Datasets:

Joemgu

/

sumstew

like 9

Summarize: As Freddie Gray was being transported in a police van through West Baltimore, at least one officer warned that Gray needed medical care but wondered, along with others, whether he was faking injuries or being uncooperative, according to investigators who reviewed the officers' statements during a departmental probe. Those statements — which have never been publicly revealed — help to explain why a judge has ordered separate trials for six officers charged in the incident. Some of the statements provide differing accounts of events that day; defense attorneys have argued in court that such conflicts could create problems in a joint trial. Officer William Porter told police investigators that after being summoned to check on Gray on the morning of April 12, he told the van's driver that the city booking facility would not process Gray because he was in medical distress. "Help me. Help me up," Gray said. Porter helped Gray up and asked, "Do you need a medic or something? Do you need to go to the hospital?" When Gray responded affirmatively, Porter said he told the van's driver, Officer Caesar Goodson, Jr., that Central Booking wouldn't accept Gray. Porter also told investigators he wasn't sure if Gray was in distress, or trying to convince officers to take him to the hospital instead of jail. That sequence of events was drawn from a police account of Porter's statement, provided as part of an initial police review of Gray's fatal injury in police custody; Goodson was the only officer charged in Gray's arrest and transport who did not provide a statement to investigators. The statements shed new light on the events of April 12 including prosecutors' allegations that officers failed to provide medical care to the 25-year-old. The Baltimore Sun was granted exclusive access to the Police Department's investigation, in which detectives outlined the officers' statements and scrutinized Gray's arrest and transport — days before any charges were filed in the case or any court proceedings began. Citizen video of Baltimore Police taking a man believed to be Freddie Gray into custody in the Gilmor Homes area on April 12. After being arrested the man was hospitalized in critical condition. A week later, he died. Citizen video of Baltimore Police taking a man believed to be Freddie Gray into custody in the Gilmor Homes area on April 12. After being arrested the man was hospitalized in critical condition. A week later, he died. SEE MORE VIDEOS Police did not show or provide The Sun with the actual statements given by the officers. Some of the officers' statements conflict, and the police summary might not reflect the full account of each officer. Those statements to investigators have become a key issue in the case. Prosecutors have asked that Porter go on trial first because they say he will be called as a witness to testify against two other officers. In a letter to Judge Barry Williams, Chief Deputy State's Attorney Michael Schatzow said Porter is "a necessary and material witness in the cases against" Goodson and Sgt. Alicia White. In an earlier hearing, defense attorneys raised concerns that the statements could create problems in a joint trial pitting one defendant's right to confront his or her accuser against another's right not to testify. Partially redacted statements by some officers were submitted to the court, though under seal. Attorneys for the officers who provided statements have argued in motions — unsuccessful to date — that the statements should be suppressed because they were not properly advised of their rights against self-incrimination or their rights under the Law Enforcement Officers' Bill of Rights. Some officers have argued that they made their statements under duress because they feared losing their jobs, or were led to believe they were providing statements as witnesses rather than suspects. Prosecutors — who allege that officers violated department policy by failing to put Gray in a seatbelt and to provide medical care — have said that all of the statements were legally and properly obtained. Gray died April 19 from an injury sustained in police custody, according to prosecutors, and his death triggered massive protests that devolved into looting and rioting. As police investigators tried to determine how Gray sustained his fatal injury, prosecutors used the local sheriff's office to conduct a parallel probe. Recently, the city agreed to pay $6.4 million to Gray's family to settle any civil claims; in that agreement, neither the city nor its officers acknowledged any wrongdoing. Goodson faces the most serious charge: second-degree murder. Porter, White and Lt. Brian W. Rice are charged with manslaughter. Officers Edward M. Nero and Garrett E. Miller face lesser charges, including second-degree assault. All have pleaded not guilty; a scheduling hearing for their trials will be held on Tuesday. Asked to comment on the officers' statements for this article, defense attorney Joseph Murtha issued a statement on behalf of all of the defense attorneys, while noting that Williams has told the prosecution and defense to refrain from discussing the case in public. "We are at a disadvantage because we cannot comment on the inaccuracy or accuracy of the statements," it said. Because police gave The Sun access to the investigation, information that "contends to represent our clients' statements is being disclosed at a time and in a manner which is both unfair and unconstitutional. We look forward to the opportunity for a complete and thorough review of all of the evidence and information that will be presented at trial." The Baltimore state's attorney's office declined to comment. Prosecutors have asked Circuit Judge Barry Williams to try William Porter first because they want to call him as a witness against Caesar Goodson and Alicia White. Porter, Goodson and White face charges of manslaughter, misconduct in office, assault and reckless endangerment. Goodson also is charged with "depraved-heart" murder. Three other officers also face charges stemming from Gray's death. The first trial is scheduled Oct. 13. However, defense attorneys indicated in a filing that some defendants might seek postponements because of "discovery issues" regarding evidence and witnesses. Williams said in a memo they would get a chance to argue for postponements. Gray's death April 19 triggered rioting in Baltimore.

Summary: An officer involved in the infamous Freddie Gray arrest-which sparked rioting and political upheaval in Baltimore after Gray died a week later-conceded that the man appeared to need medical help while in police custody, the Baltimore Sun reports. According to officer William Porter, Gray asked for help while cuffed in the back of a police van on April 12. "Help me," Gray reportedly said. "Help me up." Porter asked whether Gray needed "a medic or something," and Gray indicated yes, so Porter says he told the van's driver, officer Caesar Goodson, Jr., and Sgt. Alicia White, though he wasn't sure whether Gray was faking to avoid jail time. White checked on Gray herself, Porter says, and ordered the other officers to get the man some medical help. But they stopped to drop off another detainee at the station, where Gray was found unresponsive and taken to a hospital; there, doctors found him in cardiac arrest, perhaps with a nearly severed spine. Police provided the Sun with statements by Porter and the other officers, which sometimes contradict each other, triggering an angry response from an attorney speaking for all of the accused officers: The statements are "being disclosed at a time and in a manner which is both unfair and unconstitutional," he says. "We look forward to the opportunity for a complete and thorough review of all of the evidence and information that will be presented at trial." All of the officers have pleaded not guilty to charges including second-degree murder and manslaughter. The first of their separate trials is scheduled to start Oct. 13, the AP reports.

multi_news

en

Summarize: Neben fest und flüssig ist gasförmig einer der drei klassischen Aggregatzustände. Eine Substanz ist dann ein Gas, wenn sich ihre Teilchen in großem Abstand voneinander frei bewegen und den verfügbaren Raum gleichmäßig ausfüllen. Im Vergleich zum Festkörper oder zur Flüssigkeit nimmt die gleiche Masse als Gas unter Normalbedingungen den rund tausend- bis zweitausendfachen Raum ein. Zusammen mit den Flüssigkeiten zählen Gase zu den Fluiden. == Etymologie Die Herkunft des Wortes Gas war lange Zeit unklar. Zwar war mehr oder weniger bekannt, dass das Wort als Fachbegriff im 17. Jahrhundert durch den flämischen Arzt und Naturforscher Johan Baptista van Helmont (+ 1644) in Brüssel eingeführt wurde, über die Etymologie bestand jedoch Unsicherheit, und es wurde Herkunft u. a. aus dem Hebräischen, aus niederl. geest ("Geist"), aus niederl. gisten ("gären") oder aus deutsch gäsen (bei Paracelsus für "gären"), gäscht ("Schaum" auf gärender Flüssigkeit) vermutet. Die Klärung wurde 1859 durch den Sprachwissenschaftler Matthias de Vries herbeigeführt, der eine Aussage aus van Helmonts Ortus Medicinae (Amsterdam 1648) beibrachte, wonach dieser das Wort speziell für den durch Kälte entstandenen Dunst des Wassers bewusst neugeschaffen und hierbei eine Anlehnung an das griechische, im Niederländischen sehr ähnlich ausgesprochene Wort ao ("Chaos") bezweckt hatte: "ideo paradoxi licentia, in nominis egestate, halitum illum gas vocavi, non longe a chao veterum secretum" ("In Ermangelung eines Namens habe ich mir die Freiheit zum Ungewöhnlichen genommen, diesen Hauch Gas zu nennen, da er sich vom Chaos der Alten nur wenig unterscheidet."). == Der Aggregatzustand gasförmig Der Aggregatzustand "gasförmig" entsteht aus der "festen" oder "flüssigen" Form durch Energiezufuhr (Wärme). Für einige Elemente und Verbindungen genügen bereits die Standardbedingungen (Temperatur 20 °C, Druck 101325 Pa), um als Gas vorzuliegen; bei ausreichend hohen Temperaturen wird jedoch jede Materie in den gasförmigen Zustand versetzt. Die dabei zugeführte Energie wird zur Bewegungsenergie der einzelnen Teilchen (je nach Temperatur mit Geschwindigkeiten im Bereich um 500 m/s), was den gasförmigen Zustand mit vollständigem Ausfüllen des vorgegebenen Raumes mit statistischer Gleichverteilung der Gasteilchen bewirkt. Hierbei strebt das Gesamtsystem den Zustand höchster Entropie an (zweiter Hauptsatz der Thermodynamik). Dass dies der wahrscheinlichste Zustand ist, kann man auf folgende Weise anschaulich machen: Unterteilt man gedanklich das einem Gas zur Verfügung stehende Volumen in Raumzellen von etwa der Größe eines Moleküls, dann gibt es viel mehr Möglichkeiten, die Moleküle auf die vielen Zellen des ganzen Volumens zu verteilen als auf einen kleinen Bruchteil. Der Makrozustand der raumerfüllenden Verteilung weist die meisten Anordnungsmöglichkeiten (Mikrozustände) für die Teilchen auf und damit auch die höchste Entropie. Die Zahl der Mikrozustände, das statistische Gewicht, kann berechnet werden. Genaueres unter Entropie (Thermodynamik) /Beispiele. == Eigenschaften Bei idealen Gasen ist die freie Beweglichkeit der einzelnen Teilchen entsprechend der kinetischen Gastheorie vollkommen; dieser Zustand wird erst bei hohen Temperaturen gegenüber dem Siedepunkt erreicht (was z. B. für Wasserstoff und Helium bereits bei Zimmertemperatur gilt). Füllt man ein beliebiges ideales Gas in ein vorgegebenes Volumen, so befindet sich bei gleichem Druck und gleicher Temperatur darin immer die gleiche Anzahl Teilchen (Atome oder Moleküle), d. h. unabhängig von der Masse jedes Teilchens und somit unabhängig von der Art des Gases. Quantitativ ausgedrückt beansprucht bei Normalbedingungen ein Mol (das sind nach Avogadro 6,022 x 1023 Teilchen) jedes beliebigen Gases einen Raum von 22,4 Litern (siehe auch Molares Volumen und Loschmidt-Konstante). Bei realen Gasen sind noch mehr oder weniger große Anziehungskräfte der Teilchen untereinander wirksam (Van-der-Waals-Kräfte). Der Unterschied ist beim Komprimieren bemerkbar: Gase sind kompressibel, das Volumen idealer Gase ist umgekehrt proportional zum Druck (Zustandsgleichung). Reale Gase weichen von den vorstehend beschriebenen Gesetzmäßigkeiten mehr oder weniger ab. Gase besitzen auch Eigenschaften von Flüssigkeiten: Sie fließen und widerstehen Deformationen nicht, obgleich sie viskos sind. == Zustandsübergänge Den Übergang vom flüssigen in den gasförmigen Aggregatzustand bezeichnet man als Verdampfung (oberhalb des Siedepunktes) oder Verdunstung (unterhalb des Siedepunktes), den umgekehrten Übergang vom gasförmigen in den flüssigen Aggregatzustand als Kondensation. Der direkte Übergang vom festen in den gasförmigen Aggregatzustand ist die Sublimation, der umgekehrte Übergang vom gasförmigen in den festen Aggregatzustand heißt Resublimation. == Lagerung Um eine möglichst große Menge an Gas in einem Behälter zu speichern, also eine hohe Dichte zu erhalten, wird das Gas stark komprimiert (siehe auch Druckgas). Zwecks hoher Druckbelastbarkeit der Gasbehälter werden meist zylinderförmige oder kugelförmige Druckbehälter (z. B. Gasflaschen) eingesetzt. Gaskessel oder Gasometer sind Niederdruckspeicher mit großem geometrischen Volumen. Aufgrund des geringen Drucks (< 1 bar) ist die gespeicherte Menge aber unbedeutend. Gasversorger speichern das Gas üblicherweise im Leitungsnetz (Gasnetz), indem sie Hochdruckleitungen großer Nennweite einsetzen.

Summary: Gas nennt man einen der drei Zustände, in dem sich Materie befinden kann. Materie kann zum Beispiel Wasser oder Gold oder Sauerstoff sein. Materie ist gasförmig, fest oder flüssig. Ihr Zustand kann sich auch ändern, so kann Flüssigkeit zu Gas werden. Wasser zum Beispiel ist flüssig, zumindest, wenn es im Zimmer normal warm ist. Wenn man aber einen Topf mit Wasser auf eine heisse Herdplatte stellt, fängt das Wasser an zu kochen. Bei 100 Grad Celsius verwandelt sich das Wasser in Wasserdampf. Aus einer Flüssigkeit wird also ein Gas. Das Wasser ist verdampft. Wenn der Wasserdampf wieder kühler wird, kondensiert er, er wird also wieder zu flüssigem Wasser. Das sehen wir zum Beispiel, wenn wir unseren Atem an eine Glasscheibe hauchen. Wenn man eine bestimmte Menge Materie nimmt, dann bleibt sie in unterschiedlichen Zuständen immer gleich schwer. Als Gas braucht sie aber viel mehr Platz, sie nimmt also mehr Raum ein. Wenn man einen dichten Beutel Wasser erhitzt, platzt der Beutel schliesslich. Luft ist ein Gemisch von Gasen, vor allem von Stickstoff und Sauerstoff. Wenn Erdöl aus der Erde gewonnen wird, findet man dabei oft Erdgas. Das ist auch ein wertvoller Rohstoff, aus dem man Energie gewinnt. Helium und einige andere Gase nennt man Edelgase. So ein Edelgas verbindet sich nicht mit anderen Gasen zu neuen Gasen.

klexikon_de

de

Summarize: Following news this morning that Donald Trump ‘s star on the Hollywood Walk of Fame was smashed to pieces by an ax-wielding vandal, I couldn’t help but wonder why the face of The Apprentice has been honored with a star in the first place. The Hollywood Chamber of Commerce, which bestows the sidewalk honors, has generally not been kind to the reality TV genre. A few years ago, it made it a point to mention that it doesn’t even have a category for reality TV after Kanye West complained that his reality-star wife, Kim Kardashian, didn’t have a star. So why Trump and not Kardashian? According to Ana Martinez, the Walk of Fame’s spokeswoman, it’s not the reason you think. “He was selected for his producer job for his Miss Universe shows,” she tells me in an email. So there you have it. Trump produced beauty pageants, and that counts. Still, Trump was honored with his star in 2007, at the height of his Apprentice fame, so the timing is curious. Either way, the chamber said it will repair the star immediately. [Photo: Flickr user halbag] David Palmer says he's used to seeing unusual things on the Hollywood Walk of Fame, but what he witnesses early Wednesday caught him by surprise. Palmer describes what he saw when a man took a pickax to President Donald Trump's Walk of Fame star. (Published Wednesday, July 25, 2018) What to Know President Donald Trump's star is once again in pieces after it was smashed overnight near Hollywood and Highland by a man with a pickax The destroyed star, presented for his 'Apprentice' role in 2007, became a target for vandals during then-candidate Trump's campaign in 2016 Walk of Fame star recipients are selected by a committee that considers hundreds of applications each year, then purchased President Donald Trump's star on the Hollywood Walk of Fame was destroyed Wednesday morning by a man with a pickax that witnesses say he concealed in a guitar case, police said. Officers were called around 3:30 a.m. to the star's location on Hollywood Boulevard near Highland Avenue, where they found a small pile of rubble in place of the star that Trump received in January 2007 for his role in the NBC show "The Apprentice." The star-smashing suspect, who reported the crime to police, later turned himself in to Beverly Hills police after leaving the pickax at the scene, according to the Los Angeles Police Department. He called police and said, "See you soon," Lt. Karen Leong of the LAPD's Hollywood Division told the Los Angeles Times. The suspect was identified as Austin Clay, 24, who was booked on suspicion of felony vandalism. His hometown was not immediately available. Bond was set at $20,000, according to jail records. An initial court date was not indicated in jail records and it was not immediately clear whether he has an attorney. Witness David Palmer said he is accustomed to seeing the unusual on Hollywood Boulevard, where tourists mingle with street performers dressed as superheroes and other characters, but he was surprised to see the Walk of Fame vandalism. "I'm like, 'Why are you hitting that star? What did Donald Trump do to you?'" Palmer told NBC4. "Then he went around the corner and I think he left." Workers were sweeping up the shattered remains of the star early Wednesday. As daylight arrived, Walk of Fame visitors stopped to take pictures of the smashed star and have a closer look. One man placed an exercise pullup-bar on top of the crumbled remains, pieces of which some visitors kept as souvenirs. "We came here a few days ago and there was some grafitti on it," said tourist Josh Kallen. "We didn't think anything like this would happen." The Walk of Fame is administered by the Hollywood Chamber of Commerce, which issued a statement later Wednesday morning. Crews Clean Up Damage From Trump Walk of Fame Star Vandalism Crews clean up the damage left behind by a vandal who smashed President Donald Trump's Hollywood Walk of Fame star Wednesday July 25, 2018 with a pickax. It's expected to take several days before the star is replaced. (Published Wednesday, July 25, 2018) "The Hollywood Walk of Fame is an institution celebrating the positive contributions of the inductees," said Leron Gubler, President-CEO of the Hollywood Chamber of Commerce. "When people are unhappy with one of our honorees, we would hope that they would project their anger in more positive ways than to vandalize a California State landmark. Our democracy is based on respect for the law. People can make a difference by voting and not destroying public property." Many visitors echoed that sentiment. "I just think it's wrong," said visitor Greg Donovan, dressed in a red blazer and top hat, and carrying a "Trump 2020" sign. "If they don't like the president, they can show it in a different way." Gubler said there appears to be security camera video of the vandalism. Repairs are expected to begin immediately, but usually requires several days. During past Walk of Fame vandalism cases, stars have been covered to protect them during the work. This is at least the second time Trump's star has been vandalized in the last two years. In October 2016, a man dressed as a construction worker smashed the star with a pickax and sledgehammer. James Otis pleaded no contest to felony vandalism in February 2017 and was sentenced to three years probation, 20 days of community service and agreed to pay $4,400 for the damage. Also in 2016, the star was defaced by spray paint and even surrounded by an artist's 6-inch high wall, a miniature representation of then-candidate Trump's border wall proposal. Walk of Fame star recipients are selected by a committee that considers hundreds of applications each year. The stars are purchased for $30,000, rather than gifted. HOLLYWOOD (CBSLA) — Donald Trump’s star on the Hollywood Walk of Fame has been destroyed again, this time by a man with a pickax, who turned himself into police about an hour later. Police say the destruction was reported at about 3:30 a.m. Wednesday, and the call was believed to have been made by the man who took a pickax to the star. All that was left of the star was a jagged hole in the ground, and pieces of concrete and the star were left strewn about the sidewalk by the time police arrived. Just the star outline left of @realDonaldTrump star on #Hollywood walk of fame after man destroyed it with a pick ax early Wednesday morning @CBSLA @CBSNews pic.twitter.com/Zofgy8CU1a — JASMINE VIEL (@jasmineviel) July 25, 2018 Even in the pre-dawn hours, there were a number of witnesses to the destruction, including two security guards and a Walgreens worker who was on her way to work. “I just seen a guy going to town on, I guess, the ground or whatever,” Patricia Cox said. “I didn’t know what was going on. A guitar case was collected as evidence. It’s believed to be what held the pickax until the suspect took it out to destroy the star. And as if the destruction wasn’t complete, graffiti was added to the destroyed star then covered with a chin-up bar just a few hours later. Curious pedestrians stopped to take pictures and video of the star’s remains, and some collected pieces of it as souvenirs. A man turned himself in at the Beverly Hills Police Department at about 4:30 a.m. in connection with the destroyed star. He was later identified as Austin Clay and has been booked on suspicion of felony vandalism. Passersby seem to be fatigued with the ire being directed at the star. “Whether you like the man or not is completely inconsequential. You don’t do that to the Walk of Fame,” one man said. It was a sentiment echoed by the Hollywood Chamber of Commerce, which administers the Walk of Fame. “When people are unhappy with one of our honorees, we would hope that they would project their anger in more positive ways than to vandalize a California State landmark,” the chamber said in a statement. One man decided to stand guard over the star, holding a Trump 2020 sign. “This is the second time. Something has to be done,” Gregg Donovan said. Trump’s star had to be replaced after being vandalized in October 2016 by a man dressed as a construction worker. James Otis used a pickax and a sledgehammer then, and later pleaded no contest to felony vandalism. On Wednesday afternoon, Otis told KCAL9’s Dave Lopez he was trying to raise bail money for Clay. Since Trump announced his candidacy and his subsequent election, the star has been a magnet for visitors unhappy with the current political climate. It has been defaced by tourists and dogs, had a model of a border wall built around it, and covered with “I resist” stickers. On July 11 this year, comedian George Lopez — using a small water bottle — pretended to urinate on Trump’s star. The president received the star in 2007, while he was a producer and host of “The Apprentice.”

Summary: Vandalism has once again hit President Trump's star on the Hollywood Walk of Fame. This time, a man who had a pickax hidden in a guitar case almost completely destroyed the star, reducing it to mostly rubble, NBC Los Angeles reports. The suspect left the pickax at the star and reported the incident to police himself, around 3:30am Wednesday, and later turned himself in. Graffiti was later added to the already-destroyed star, CBS Los Angeles reports. In 2016, a man used a pickax and sledgehammer to deface the star of then-candidate Trump (who received it in 2007 for his role as the Miss Universe producer, Fast Company explained in 2016). It has also been spray-painted, covered with stickers, and had a 6-inch-high wall placed around it to symbolize Trump's proposed border wall. It's not clear when repairs will begin this time around. See the destroyed star here.

multi_news

en

Write a title and summarize: SECTION 1. SHORT TITLE. This Act may be cited as the ``National Veterinary Medical Service Act''. SEC. 2. ESTABLISHMENT OF LOAN REPAYMENT PROGRAM FOR VETERINARY MEDICINE. The National Agricultural Research, Extension, and Teaching Policy Act of 1977 is amended by inserting after section 1415 (7 U.S.C. 3151) the following: ``SEC. 1415A. VETERINARY MEDICINE LOAN REPAYMENT. ``(a) Definitions. ``In this section: ``(A) Qualifying educational loan.--The term `qualifying educational loan' means a government or commercial loan incurred by an individual to pay for attendance at an accredited college of veterinary medicine resulting in a degree of Doctor of Veterinary Medicine or the equivalent, including-- ``(i) tuition expenses; ``(ii) other reasonable educational expenses, including fees, books, and laboratory expenses; and ``(iii) reasonable living expenses, as determined by the Secretary. ``(B) Veterinarian shortage situation.-- ``(i) In general.--The term `veterinarian shortage situation' shall have the meaning that the Secretary shall prescribe. ``(ii) Considerations.--In determining what constitutes a veterinarian shortage situation, the Secretary may consider-- ``(I) urban or rural areas that the Secretary determines have a shortage of veterinarians;---------- ``(II) areas of veterinary practice that the Secretary determines have a shortage of veterinarians, such as public health, epidemiology, and food safety; ``(III) areas of veterinary need in the Federal Government; and ``(IV) other factors that the Secretary considers to be relevant. ``(b) Program.-- ``(1) Service in veterinarian shortage situations.-- ``(A) In general.--Subject to the availability of funds under subsection (d), the Secretary shall carry out a program to enter into agreements with veterinarians under which a veterinarian agrees to provide, for a period of time determined by the Secretary and specified in the agreement, veterinary services in veterinarian shortage situations. ``(B) Annual loan repayment.--For each year of service under an agreement under this paragraph, the Secretary shall pay an amount, determined by the Secretary and specified in the agreement, of the principal, interest, and related expenses of the qualifying educational loans of the veterinarian. ``(2) Service to federal government in emergency situations.--The Secretary may enter into a 1-year agreement with a veterinarian who has entered into an agreement under paragraph (1) for the veterinarian to provide service to the Federal Government in an emergency situation, as determined by the Secretary, under terms and conditions specified in the agreement. ``(C) Additional annual loan repayment.-- ``(i) In general.--Under an agreement under this paragraph, the Secretary shall pay an amount, as determined by the Secretary and specified in the agreement, of the principal, interest, and related expenses of the qualifying educational loans of the veterinarian. ``(ii) Amount in addition.--An amount paid under clause (i) shall be in addition to the amount paid under the agreement under paragraph (1). ``(D) Requirements.--An agreement under this paragraph shall provide that-- ``(i) a veterinarian shall not be required to serve more than 60 working days per year of the agreement; and ``(ii) a veterinarian who provides service in accordance with the agreement-- ``(I) shall receive a salary commensurate with the duties performed; and ``(II) shall be reimbursed for travel and per diem expenses as appropriate for the duration of the service. ``(c) Administration.-- ``(1) Authority.--The Secretary may carry out the program under subsection (b) directly or through an agreement with another Federal agency or service provider. ``(2) Breach remedies.-- ``(A) In general.--An agreement under paragraph (1) or (2) of subsection (b) shall provide remedies for a breach of the agreement by the veterinarian, including repayment or partial repayment of financial assistance received, with interest. ``(B) Amounts recovered.--Funds recovered under this subsection-- ``(i) shall be credited to the account available to carry out this section; and ``(ii) shall be available to carry out this section without further appropriation until expended. ``(C) Waiver.--If the Secretary determines that a veterinarian has breached an agreement due to extreme hardship or extreme need, the Secretary may grant a waiver of the repayment obligation for breach of contract. ``(4) Repayment schedule.--The Secretary may enter into an agreement with the holder of a qualifying educational loan for which payments are made under this section to establish a schedule for the making of payments. ``(5) Tax liability.--In addition to educational loan repayments, the Secretary shall make such additional payments to a participating veterinarian as the Secretary determines to be appropriate to reimburse the veterinarian for tax liability resulting from participation in the program under this section. ``(d) Authorization of Appropriations.--There are authorized to be appropriated such sums as are necessary to carry out this section, to remain available until expended.''.

Title: A bill to authorize the Secretary of Agriculture to conduct a loan repayment program to encourage the provision of veterinary services in shortage and emergency situations Summary: National Veterinary Medical Service Act - Amends the National Agricultural Research, Extension, and Teaching Policy Act of 1977 to direct the Secretary of Agriculture to conduct a veterinary school loan repayment program (including resultant tax liability payments) for persons who perform qualifying veterinary services in shortage and emergency situations.

billsum

en

Summarize: FILE - In this June 26, 2012, file photo, two women converse in New York. "Socially accepted normal body weight is shifting toward heavier weight. As more people around us are getting heavier, we simply... (Associated Press) FILE - In this June 26, 2012, file photo, two women converse in New York. "Socially accepted normal body weight is shifting toward heavier weight. As more people around us are getting heavier, we simply believe we are fine, and no need to do anything with it," said Dr. Jian Zhang, a public health researcher... (Associated Press) FILE - In this June 26, 2012, file photo, two women converse in New York. "Socially accepted normal body weight is shifting toward heavier weight. As more people around us are getting heavier, we simply believe we are fine, and no need to do anything with it," said Dr. Jian Zhang, a public health researcher... (Associated Press) FILE - In this June 26, 2012, file photo, two women converse in New York. "Socially accepted normal body weight is shifting toward heavier weight. As more people around us are getting heavier, we simply... (Associated Press) CHICAGO (AP) — Fewer overweight Americans have been trying to lose weight in recent years, and researchers wonder if fat acceptance could be among the reasons. The trend found in a new study occurred at the same time obesity rates climbed. "Socially accepted normal body weight is shifting toward heavier weight. As more people around us are getting heavier, we simply believe we are fine, and no need to do anything with it," said lead author Dr. Jian Zhang, a public health researcher at Georgia Southern University. Another reason could be people abandoning efforts to drop pounds after repeated failed attempts, Zhang said. The researchers analyzed U.S. government health surveys over nearly two decades from 1988 through 2014. The surveys involved in-person physical exams and health-related questions including asking participants if they'd tried to lose weight within the past year. More than 27,000 adults aged 20 to 59 were included. They were not asked to explain their answers. In the early surveys, about half the adults were overweight or obese. Those numbers climbed to 65 percent by 2014. But the portion of overweight or obese adults who said they were trying to slim down fell from 55 percent to 49 percent in the study. Body mass index, a measure of height and weight, determines weight status. Those with a BMI of 25 to 29 are considered overweight; 30 and above is obese. A BMI of 30 generally reflects being about 50 pounds above your ideal weight. The study results were published Tuesday in the Journal of the American Medical Association. Dr. Scott Kahan, director of a weight-loss clinic in Washington, said the study is important and echoes previous research. He acknowledged that it has become more acceptable in some circles to be overweight, but that many patients still feel stigmatized. He said many come to his center after repeated attempts to lose weight and some give up for a while out of frustration. The study found obesity was most common among black women — 55 percent were obese in the most recent survey years, and there was a big decline in black women trying to lose weight. Whether that's because of fat acceptance, dieting frustration or other reasons is not known. Zhang said there's a positive side to fat acceptance, if it means people feel less ridiculed for their weight. But obesity can increase risks for heart disease, diabetes, cancer and other ailments. The findings "are a very serious concern," he said. "We should forget the words 'fat' or 'obesity,'" Zhang said, adding that a healthy lifestyle may be an effective way to help people lose weight. ___ Online: CDC: https://www.cdc.gov/obesity/ ___ Follow AP Medical Writer Lindsey Tanner at http://www.twitter.com/LindseyTanner. Her work can be found at http://bigstory.ap.org/content/lindsey-tanner What is BMI? BMI is a person’s weight in kilograms divided by the square of height in meters. BMI does not measure body fat directly, but research has shown that BMI is moderately correlated with more direct measures of body fat obtained from skinfold thickness measurements, bioelectrical impedance, densitometry (underwater weighing), dual energy x-ray absorptiometry (DXA) and other methods 1,2,3. Furthermore, BMI appears to be as strongly correlated with various metabolic and disease outcome as are these more direct measures of body fatness 4,5,6,7,8,9. In general, BMI is an inexpensive and easy-to-perform method of screening for weight category, for example underweight, normal or healthy weight, overweight, and obesity. How is BMI used? A high BMI can be an indicator of high body fatness. BMI can be used as a screening tool but is not diagnostic of the body fatness or health of an individual. To determine if a high BMI is a health risk, a healthcare provider would need to perform further assessments. These assessments might include skinfold thickness measurements, evaluations of diet, physical activity, family history, and other appropriate health screenings10. What are the BMI trends for adults in the United States? The prevalence of adult BMI greater than or equal to 30 kg/m2 (obese status) has greatly increased since the 1970s. Recently, however, this trend has leveled off, except for older women. Obesity has continued to increase in adult women who are age 60 years and older. To learn more about the trends of adult obesity, visit Adult Obesity Facts. Top of Page Why is BMI used to measure overweight and obesity? BMI can be used for population assessment of overweight and obesity. Because calculation requires only height and weight, it is inexpensive and easy to use for clinicians and for the general public. BMI can be used as a screening tool for body fatness but is not diagnostic. To see the formula based on either kilograms and meters or pounds and inches, visit How is BMI calculated? Top of Page What are some of the other ways to assess excess body fatness besides BMI? Other methods to measure body fatness include skinfold thickness measurements (with calipers), underwater weighing, bioelectrical impedance, dual-energy x-ray absorptiometry (DXA), and isotope dilution 1,2,3. However, these methods are not always readily available, and they are either expensive or need to be conducted by highly trained personnel. Furthermore, many of these methods can be difficult to standardize across observers or machines, complicating comparisons across studies and time periods. Top of Page How is BMI calculated? BMI is calculated the same way for both adults and children. The calculation is based on the following formulas: Measurement Units Formula and Calculation Kilograms and meters (or centimeters) Formula: weight (kg) / [height (m)]2 With the metric system, the formula for BMI is weight in kilograms divided by height in meters squared. Because height is commonly measured in centimeters, divide height in centimeters by 100 to obtain height in meters. Example: Weight = 68 kg, Height = 165 cm (1.65 m) Calculation: 68 ÷ (1.65)2 = 24.98 Pounds and inches Formula: weight (lb) / [height (in)]2 x 703 Calculate BMI by dividing weight in pounds (lbs) by height in inches (in) squared and multiplying by a conversion factor of 703. Example: Weight = 150 lbs, Height = 5’5″ (65″) Calculation: [150 ÷ (65)2] x 703 = 24.96 Top of Page How is BMI interpreted for adults? For adults 20 years old and older, BMI is interpreted using standard weight status categories. These categories are the same for men and women of all body types and ages. The standard weight status categories associated with BMI ranges for adults are shown in the following table. BMI Weight Status Below 18.5 Underweight 18.5 – 24.9 Normal or Healthy Weight 25.0 – 29.9 Overweight 30.0 and Above Obese For example, here are the weight ranges, the corresponding BMI ranges, and the weight status categories for a person who is 5′ 9″. Height Weight Range BMI Weight Status 5′ 9″ 124 lbs or less Below 18.5 Underweight 125 lbs to 168 lbs 18.5 to 24.9 Normal or Healthy Weight 169 lbs to 202 lbs 25.0 to 29.9 Overweight 203 lbs or more 30 or higher Obese Top of Page For children and teens, the interpretation of BMI depends upon age and sex. For more information about interpretation for children and teens, read – What is a BMI percentile and how is it interpreted? Is BMI interpreted the same way for children and teens as it is for adults? BMI is interpreted differently for children and teens, even though it is calculated using the same formula as adult BMI. Children and teen’s BMI need to be age and sex-specific because the amount of body fat changes with age and the amount of body fat differs between girls and boys. The CDC BMI-for-age growth charts take into account these differences and visually show BMI as a percentile ranking. These percentiles were determined using representative data of the U.S. population of 2- to 19-year-olds that was collected in various surveys from 1963-65 to 1988-9411. Obesity among 2- to 19-year-olds is defined as a BMI at or above the 95th percentile of children of the same age and sex in this 1963 to 1994 reference population. For example, a 10-year-old boy of average height (56 inches) who weighs 102 pounds would have a BMI of 22.9 kg/m2. This would place the boy in the 95th percentile for BMI – meaning that his BMI is greater than that of 95% of similarly aged boys in this reference population – and he would be considered to have obesity. For more information and to access the CDC Growth Charts For adults, the interpretation of BMI does not depend on sex or age. Read more about interpreting adult BMI. Top of Page How good is BMI as an indicator of body fatness? The correlation between the BMI and body fatness is fairly strong1,2,3,7, but even if 2 people have the same BMI, their level of body fatness may differ12. In general, At the same BMI, women tend to have more body fat than men. At the same BMI, Blacks have less body fat than do Whites 13,14, and Asians have more body fat than do Whites 15, and Asians have more body fat than do Whites At the same BMI, older people, on average, tend to have more body fat than younger adults. At the same BMI, athletes have less body fat than do non-athletes. The accuracy of BMI as an indicator of body fatness also appears to be higher in persons with higher levels of BMI and body fatness16. While, a person with a very high BMI (e.g., 35 kg/m2) is very likely to have high body fat, a relatively high BMI can be the results of either high body fat or high lean body mass (muscle and bone). A trained healthcare provider should perform appropriate health assessments in order to evaluate an individual’s health status and risks. Top of Page If an athlete or other person with a lot of muscle has a BMI over 25, is that person still considered to be overweight? According to the BMI weight status categories, anyone with a BMI between 25 and 29.9 would be classified as overweight and anyone with a BMI over 30 would be classified as obese. However, athletes may have a high BMI because of increased muscularity rather than increased body fatness. In general, a person who has a high BMI is likely to have body fatness and would be considered to be overweight or obese, but this may not apply to athletes. A trained healthcare provider should perform appropriate health assessments in order to evaluate an individual’s health status and risks. Top of Page What are the health consequences of obesity for adults? People who have obesity are at increased risk for many diseases and health conditions, including the following: 10, 17, 18 All-causes of death (mortality) High blood pressure (Hypertension) High LDL cholesterol, low HDL cholesterol, or high levels of triglycerides (Dyslipidemia) Type 2 diabetes Coronary heart disease Stroke Gallbladder disease Osteoarthritis (a breakdown of cartilage and bone within a joint) Sleep apnea and breathing problems Chronic inflammation and increased oxidative stress 19,20 Some cancers (endometrial, breast, colon, kidney, gallbladder, and liver) Low quality of life Mental illness such as clinical depression, anxiety, and other mental disorders 21,22 Body pain and difficulty with physical functioning23 For more information about these and other health problems associated with obesity, visit Health Effects Top of Page Socially acceptable body weight is increasing.1 If more individuals who are overweight or obese are satisfied with their weight, fewer might be motivated to lose unhealthy weight. This study assessed the trend in the percentage of adults who were overweight or obese and trying to lose weight during 3 periods from 1988 through 2014. We used data from the National Health and Nutrition Examination Survey (NHANES), an ongoing, stratified, multistage probability sample of the US noninstitutionalized population designed to represent the health and nutritional status of the general population. A strength of NHANES is that the sampling approaches, interviews, and physical examination methods are standardized across surveys and have been published extensively elsewhere.2 NHANES protocol was approved by the National Center for Health Statistics institutional review board, and written informed consent was obtained.2 The current analysis was categorized as exempt by the Georgia Southern University institutional review board. Although weight gain has continued among U.S. adults, fewer report trying to lose weight, according to a study appearing in the March 7 issue of JAMA. Socially acceptable body weight is increasing. If more individuals who are overweight or obese are satisfied with their weight, fewer might be motivated to lose unhealthy weight. Jian Zhang, M.D., Dr.P.H., of Georgia Southern University, Statesboro, and colleagues used data from the National Health and Nutrition Examination Survey (NHANES) to assess the trend in the percentage of adults who were overweight or obese and trying to lose weight during three periods: from 1988-1994, 1999-2004, and 2009-2014. Participants ages 20 to 59 years who were overweight (a body mass index [BMI] of 25 to less than 30) or obese (BMI 30 or greater) were included. The question of interest was "During the past 12 months, have you tried to lose weight?" The study included 27,350 adults. Overweight and obesity prevalence increased throughout the study period, from 53 percent in 1988-1994 to 66 percent in 2009-2014. The percentages of adults who were overweight or obese and trying to lose weight declined during the same period, from 56 percent in 1988-1994 to 49 percent in 2009-2014. The largest decline occurred among black women, from 66 percent in 1988-1994 to 55 percent in 2009-2014. Black women also had the highest prevalence of obesity, and more than half of black women (55 percent) were obese in the 2009-2014 survey. Adjusted prevalence rates showed a significantly declining trend of reporting efforts to lose weight among white men and women, and black women. The authors write that fewer adults trying to lose weight may be due to body weight misperception reducing the motivation to engage in weight loss efforts, or primary care clinicians not discussing weight issues with patients. Also, the longer adults live with obesity, the less they may be willing to attempt weight loss, in particular if they had attempted weight loss multiple times without success. Explore further: Overweight mothers underestimate their children's weight More information: JAMA. DOI: 10.1001/jama.2016.20036

Summary: Despite a steady rise of obesity rates, fewer overweight Americans are trying to drop extra pounds, new research shows. "Socially acceptable body weight is increasing," the scientists write in JAMA. They theorize that as individuals see more overweight people around them, they become less "motivated to lose unhealthy weight." Another reason could be the demoralizing effect of failed diets, lead author Dr. Jian Zhang tells the AP. Zhang's team studied government health data stretching back 20 years for 27,000 adults aged 20 to 59. The surveys involved a physical exam and this question: "During the past 12 months, have you tried to lose weight?" About half of the adults were overweight or obese at the start with a body mass index of 25 to above 30. A BMI of 30 or above is considered obese, per the CDC. The percentage of those overweight or obese rose throughout the study period from 53% to 66% by the end in 2014, per Medical Xpress. At the same time, the percentage of those trying to shed pounds dropped from 56% to 49%. Black women had the highest obesity rates and the sharpest drop in a desire to lose pounds, from 66% to 55%. While there is a bright side to fat acceptance, Zhang says the findings are a "serious concern," with obesity linked to serious health effects such as heart disease and cancer. Weight loss is "really challenging," says Zhang, per the Daily News. "Let's forget 'fat,' 'obese,' all the bad words.... Let's just try to engage in a healthy lifestyle: eating less and moving."

multi_news

en

Summarize: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention generally relates to training aids for use in sporting activities. More particularly, the present invention is directed to an improved method and apparatus for instructing a batter on the proper technique for hitting a baseball or softball with a bat. 2. Description of the Prior Art and Related Information It has been said that many professional athletes in all different sports consider the art of hitting a baseball or softball correctly to be the single most difficult thing to do in any sport. In little league baseball, a batter has one to two seconds to identify a pitch, make a decision whether to swing at a pitch, and then complete a proper swing of the bat to make contact with the ball. In professional baseball, where it is not uncommon to have pitchers who can throw a baseball between 80 and 100 miles per hour, a batter has one-fifth of a second (0.2 seconds) to identify the pitch, decide whether to swing the bat, and to swing the bat so as to make contact with the ball! In addition, given that the useful area of bat for making proper contact with the ball is approximately four to six inches long and about one and one-half inches wide (depending on the location of the pitch and size of the bat), one can readily understand the difficulty associated with becoming a successful batter. Perhaps this explains why, in professional baseball, players are considered good hitters if they are able to successfully hit a baseball as few as 25 to 30 times out of 100 attempts. It also helps explain why Ted Williams, who in one memorable season successfully hit the ball an average of more than 40 times per 100 attempts, is considered in mythic proportions. Very few believe his achievement will ever be duplicated. Given the love for baseball in the United States, it is not to be unexpected to find prior art related to the devices for attempting to teach proper batting techniques. The most common prior art batting instruction aid in use today is a standard batting tee in which a tube, oftentimes adjustable in height, is attached to a piece or rubber or plastic in the shape of a home plate, and a ball is supported by the tube. Such a batting tee is used to simulate the ball, having been pitched by a pitcher, crossing the plate for the batter to swing at. However, the standard batting tee described above, which is used in little leagues, grade schools, high schools, colleges, and by professional baseball and softball teams across the country, has a major flaw. That is, depending upon the location of the pitch which is thrown, it may be necessary for the batter to swing at and make contact with the ball either before the ball reaches the plate or after the ball has passed the plate and is approaching the glove of the catcher. By &#34;location,&#34; it is meant that a ball may be thrown by the pitcher inside, outside, high or low. Such pitches may be thrown in or out of the strike zone, and the location of the pitch, i.e., inside, outside, etc., will, to a great extent, dictate the batter&#39;s swing. For example, for an inside pitch, it is necessary for the hitter to begin swinging the bat and make contact with the ball when the ball is well in front of the plate. Similarly, for an outside pitch, in order to make proper contact with the ball, the batter must swing the bat and make contact with the ball as the ball is crossing the back threshold of the plate. This is commonly referred to a &#34;going with the pitch.&#34; However, prior art batting tees teach a batter to swing at a ball so as to contact the ball as it is crossing the center of the plate. As discussed above, this is not proper form, depending on the location of the pitch. Thus, prior art batting tees teach a batter what is essentially an incorrect method for the development of proper batting technique, particularly for younger players. &#34;Tee-ball&#34; leagues for young children have sprung up all over the United States and in many foreign countries. This type of a league utilizes a batting tee instead of having a pitcher pitch the ball. In such a league, the children are very young and are developing the fundamental skills necessary to play the game. It is therefor essential that these children are taught proper batting technique. However, the use of a standard batting tee discussed above teaches such children to hit a baseball or softball as the ball is crossing the center of the plate. This is incorrect. As an alternative, some coaches try to improve upon the prior art batting tee by using a batting tees placed in front of home plate. The ball is placed on the tee and the children are instructed to align themselves with the actual home plate behind the plate which is attached to the batting tee. While this technique is somewhat better than using only the batting tee itself, children often have difficulty transferring the lessons learned from this type of instruction method to an actual playing situation in which there is only the batting tee. In this situation, children can become confused, discouraged and disheartened with the game of baseball. While it has often been said that practice makes perfect, if a player does not practice using the proper technique, the player will ultimately perfect what are essentially bad habits conducive to poor hitting performance. Once an individual has learned an improper technique for hitting a baseball or softball, it is extremely difficult for such an individual to rid themselves of the bad habits developed through the improper learning technique. Accordingly, there is a need for a device which properly simulates a pitch thrown by a pitcher such that the batter can learn when to swing at a ball depending upon the location of the pitch and can also learn where to stand relative to the plate to achieve the best results. In addition, there is a need for a method and apparatus for teaching the proper fundamentals of batting such as alignment with home plate and the proper contact points between the bat and the ball to ensure the development of a good batting technique. SUMMARY OF THE INVENTION The present invention is directed to an improved batting instruction method and apparatus in which batters are taught proper batting technique for any location where a pitch may be thrown by a pitcher. In addition, batters are taught proper placement of themselves relative to the actual plate and when to swing at a ball depending upon the location of the pitch. With the present invention, an individual will learn proper batting technique and avoid the pitfalls associated with utilizing prior art devices. The present invention obviates the problems associated with the prior art using a unique form of batting tee that allows the ball to be properly placed, relative to home plate, for hitting depending on the location of the pitch being simulated. Further, the present invention utilizes indicia for indicating proper contact between the bat and the ball. Using the present invention, a batter sets up relative to home plate and a ball can be positioned at an optimum hitting point relative to the plate. Thus, the batter learns proper stance relative to home plate as well as where to swing at the ball in order to make proper contact depending on the location of the pitch. The present invention, which has been generally discussed above, will be more readily understood through the accompanying detailed description of the invention taken in conjunction with the figures of which the following is a brief description: BRIEF DESCRIPTION OF THE FIGURES FIG. 1 shows a perspective view of the batting instruction device of the present invention; FIG. 2 shows a side elevational view of the batting instruction device of FIG. 1; FIG. 3 shows a detail of the connection between the plate and the ball support mechanism of FIG. 1; FIG. 4 shows a detail of the fit between various portions of the ball supporting device of FIG. 1; FIG. 5 shows a bat in accordance with the present invention for use with the device shown in FIG. 1; FIG. 6 shows a ball for use in conjunction with the device of FIG. 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT As can seen in FIGS. 1-4, the present invention includes a home plate 10 having a first ball supporting device connected thereto. The home plate 10 is provided with a plurality of openings 24 formed therein so that the ball supporting devices can be attached to various positions about the plate. The first ball supporting device includes a first vertical member 12. The first vertical member 12 extends upward therefrom and mates with a radial member 14. The radial member 14 mates with a second vertical member 16. A third vertical member 18, which supports a baseball 50 on one end thereof mates with the second vertical member 16. A support 26, which can be formed to fit about the circumference of the radial member 14, is provided to prevent excessive sagging of the radial member 14 due to the weight of vertical members 16 and 18. A second ball supporting device can be connected to the home plate 10 at the same time as the first ball supporting device. The second support device is shown comprised of vertical members 20, 21, 22, and 23 which are similar in construction to the above discussed vertical and radial members. By making the second ball support device with four interconnecting members, a wide range of height adjustment can be provided to simulate multiple pitches at various points in the strike zone. An advantage to using two ball supporting devices together will become clear from the discussion below. The vertical and radial members described above are preferably formed of a heavy duty rubber material, which can be easily molded using a simple molding process. Other suitable, stiff yet pliable materials could also be used with the present invention. Alternatively, it would be possible to make several of the components out of different materials, such as metal. That is, while it is necessary to have the vertical member 18 (or possibly vertical member 16 depending the particular use to which the present invention is put) which supports the baseball be made of a flexible material so that when a batter swings the bat and hits the vertical member 18, it does not break, the vertical portion 12 and the radial portion 14 could conceivably be made of a less resilient material provided that it is rugged. For example, a heavy duty plastic could be appropriate. Support 26 can also be made of metal, rubber or plastic and can also be formed as a single extension extending down from the radial member 14 as opposed to the bi-pod structure shown in the drawings. Members 12, 14, 16, and 18 can be formed of any appropriate size. Preferably, vertical member 12 extends 10&#34; above the plate, 18&#34; along the horizontal plane of the plate, and is 2.5&#34; in diameter. Radial member 14 has a length of 18&#34; along the horizontal and vertical portions and a diameter of approximately 2.25&#34;. Vertical member 16 has a length of 20&#34; and a diameter of approximately 2&#34;. Vertical member 18 has a length of 20&#34; and a diameter of 1.75&#34;. Vertical members 20, 21, 22, and 23 which make up a second ball support for use with the present invention, are preferably sized at 10&#34; by 2.5&#34;, 20&#34; by 2.25&#34;, 20&#34; by 2&#34;, and 20&#34; by 1.75&#34;, respectively. In the preferred embodiment, vertical members 12, 16, and 18 and radial member 14 fit together utilizing a friction fit which is shown in more detail in FIG. 4. In particular, the fit between the various members may be a &#34;telescoping&#34; fit in which the individual members slide together in a coaxial fashion with friction holding the individual members at desired positions. To assist in making a proper and secure friction fit, the outer coaxial member can be provided with grooves or ridges 28 formed therein. The plurality of grooves or ridges 28 surrounding the inserted member serve to provide sufficient friction to hold the inserted member in the desired position yet allow the relative positions of the connected members to be adjusted by twisting or otherwise pulling the inserted member with sufficient force to overcome the friction force created by the grooves or ridges 28. Referring in particular to FIG. 3, the interconnection of the vertical member 12 and the plate 10 is illustrated. A bolt 30 having a head 32 and a threaded portion 34 is inserted through an opening 24 in the plate 10 and interconnects with a threaded coupling 36 disposed in the vertical member 12. Threaded coupling 36 is preferably made of metal to provide a secure connection. In addition, coupling 36 can extend up into the vertical member 12 any desired length to assist in providing structural rigidity to the ball support device. As shown, the coupling is disposed in the vertical member 12 which can comprise a heavy duty rubber which is molded about the coupling 36. Alternatively, the vertical member 12 can be made entirely of metal to provide additional structural integrity. The bolt is tightened and the vertical member 12 is held securely to the plate. The plate 10 can include molded detents on the underside of the plate 10 which are formed to fit around the head portion 32 of bolt 30. Thus, the bolt can be held in place while the vertical member 12 including the coupling 36 is twisted about the threaded portion 34 to secure the member 12 to the plate 10. Due to the threaded interconnection of the coupling 36 and the threaded portion 34, the ball supporting member can be swiveled about the plate 10 to properly position the ball supporting device to simulate a desired pitch. In addition, the telescoping connections between members 12, 14, 16 and 18 allow an almost limitless variation of the ball position. The coupling 36 and the bolt 30 are preferably made of metal. Alternative methods for connecting the ball supporting devices to the plate 10 are intended to fall within the scope of this invention. In particular, it is clear that the bolt 30 could be permanently attached to the vertical member 12 and extend through the top of plate 10 and protrude from the bottom thereof and be secured by way of a nut. Alternatively, the vertical member 12 could be provided with a keyed member protruding therefrom which is inserted into a pre-formed &#34;key-hole&#34; opening in the plate 10. After insertion, the vertical member 12 could be turned to securely hold the vertical member 12 to the plate 10. Referring to FIGS. 5 and 6, the bat 40 is provided with distinct hit indicators 42 and 44. The ball 50 is provided with distinct strike indicators 52 and 54. In practice, hit indicator 42 corresponds to strike indicator 52 and hit indicator 44 corresponds to strike indicator 54. That will become more clear from the discussion below as to the use of the present invention. The size of the hit indicators 42, 44 are determined by measuring from the end of the bat 48 farthest from the handle 46 approximately 10 inches down the length of the shaft of the bat. At the measured distance, a line is drawn about the circumference of the bat. A second measurement of approximately three inches is taken from the end 48 toward the handle 46. A second line is drawn about the circumference of the bat. The two lines define a cylindrically shaped portion 49 of the bat which is approximately seven inches long. Depending on the size of the bat, this portion may vary in size (larger or smaller). The cylindrical portion 49 is then divided in half along the longitudinal axis thereof and only one half of the cylinder is utilized (either side). If a wooden bat is utilized, the division is taken, preferably, along a longitudinal axis which, if extended, would split the manufacturer&#39;s label 41 in half. The half cylinder portion is again divided in half along the longitudinal axis of the bat to produce the areas defining the hit indicators 42, 44. These areas are then marked to distinguish them from the rest of the bat. E.g., one section 42 could be painted blue, while the other 44 could be painted red. The process of providing strike indicators 52, 54 for the ball 50 is somewhat less complicated. Simply put, the ball is divided into two equal halves 52, 54, which are then marked to correspond to the hit indicators 42, 44. Although corresponding markings are not absolutely necessary, it is helpful to avoid confusion. Thus, if hit indicator 42 is painted red, strike indicator 52 would be painted red as well. Similarly, if hit indicator 44 is painted blue, strike indicator 54 would be painted blue. The operation of the present invention will now be described. Referring to FIG. 1, which is set up for left-handed batter, vertical member 23 supports a ball 50 to simulate an outside pitch. The ball supporting device including members 12, 14, 16 and 18 would simulate an inside pitch. Prior to swinging, using the bat shown in FIG. 5, the batter would stand at the plate 10 (in a proper batting stance relative to plate 10) and hold the bat such that the label 41 thereon is facing straight up (in the opposite direction from the plate) and the hit indicators 42, 44 are facing towards the forward portion of the plate 10. Further, balls 50 are positioned on the members 23, 18 such that the strike indicator 54 corresponding to the top hit indicator 44 (when bat 40 is held out over the plate) faces away from the batter and the centerline 53 of ball 50 is substantially parallel to the lateral side edge 11 of the plate 10. At this point, the batter swings the bat 40 so as to make contact with ball 50 supported by member 23.and ball 50 supported by member 18. However, when bat 40 contacts ball 50 supported by the member 23 to simulate an outside pitch, the batter needs to utilize that portion of the bat 40 identified as hit indicator 42. Furthermore, hit indicator 42 must meet the ball 50 along the strike portion 52. In this manner, the outside pitch will be hit to the opposite field, in this case to the left field. As the batter continues swinging the bat 40, proper hand rotation will cause that portion of the bat 40 identified as hit indicator 44 to contact ball 50 supported by vertical member 18. Hit indicator 44 should make contact with strike indicator 54 on the ball 50. In this manner, the ball 50 will be pulled to the right field which is proper for an inside pitch. As evident from a review of FIG. 1, ball 50 supported by vertical member 23, which simulates an outside pitch, is properly struck when the ball 50 has passed the majority of the plate 10. Conversely, ball 50 supported by vertical member 18 and simulating an inside pitch, must be struck before the ball reaches the plate 10 in order to be hit properly. By controlling the swing of the bat 40 to ensure that hit indicator 42 contacts the ball 50 on member 23 on the strike indicator 52 and hit indicator 54 contacts ball 50 on member 18 on the strike indicator 54, the batter will ensure proper swing technique and wrist release. By repeatedly practicing this motion, the batter will develop proper batting technique. The above discussion illustrates that a proper batting technique will result in the same swing to make contact with an outside pitch as with an inside pitch. Given the speed at which the ball 50 is often travelling in actual game situations, this technique teaches the batter that he or she must anticipate an outside pitch in order to correctly hit an inside pitch. Since the present invention is able to simulate both inside and outside pitches at the same time, a batter can train to hit the outside ball to the opposite field, and following through, pull the inside pitch down the line to the proper field (for the left-handed batter set up of FIG. 1 this would be the right field). Finally, the present invention provides instant feedback as to whether a batter is making proper contact with ball 50. A coach can monitor the performance of a player and determine whether the player is making proper contact between the bat 40 and the ball 50 depending upon which portion of the bat 40 contacts the ball 50. Using the above-described method and apparatus for teaching batting technique, a batter will develop the proper fundamental skills on how to properly hit a baseball. Further, an individual will not only learn proper batting technique, but also proper placement of his or her body relative to the plate 10 shown in FIG. 1. There is no need to use multiple batting tees to teach proper position relative to the plate as with the prior art or to move the ball supporting device and plate to properly position a ball since the present invention accurately positions the ball depending upon the location of pitch the being simulated. Accordingly, the present invention overcomes the problems associated with prior art batting tees and teaches individuals of all ages proper batting techniques.

Summary: A baseball instructional device for use teaches a hitter how to properly hit a ball depending on the location at which the ball is thrown by the pitcher. The device is capable of positioning a baseball, softball, or other ball, at proper hitting positions relative to the hitter, thus teaching the hitter when and where to swing to make proper contact with the ball. In addition to positioning the ball relative to the hitter, the device is provided with visual indicators to assist in teaching the hitter the proper location for striking the ball with the bat.

big_patent

en

Summarize: By. Liz Hull. PUBLISHED:. 04:58 EST, 15 January 2014. |. UPDATED:. 18:09 EST, 15 January 2014. A woman struggling to cope after breaking up with her boyfriend went berserk and strangled her mother, a court heard. Emma Parr, 38, transformed from a ‘happy, bubbly young woman’ to a killer in the space of just six months. She was suffering from severe depression when she throttled her mother Carol, 62, who had told her that she was planning to sell the family home where they lived happily together. Parr recollected little of what had happened though she spoke of disappointment her mother no longer wanted to live with her. Emma Parr, 38, turned on waitress Carol Parr, 62, following her struggle to cope with the bitter break up. Mrs Parr’s body was discovered by a relative after colleagues raised the alarm when she failed to turn up for work. Police. later found her daughter, who had attempted suicide just weeks earlier,. driving her car in her pyjamas 17 miles away. When questioned, the. university department manager told police she remembered little of what. had happened, except ‘disappointment’ that her mother had decided she. didn’t want to live with her. Psychiatric. examinations concluded Miss Parr was suffering from a ‘severe. depressive episode’ and had psychotic symptoms at the time. Yesterday. she was jailed for just ten months after a judge ruled she was a. ‘caring and loving person’ whose deadly attack was ‘largely outside her. control’. She admitted manslaughter on the grounds of diminished. responsibility after the prosecution accepted her not guilty plea to. murder. Mr Justice. Alistair MacDuff told Miss Parr: ‘This is the saddest of cases. Just one. short year ago you were a happy bubbly young woman with many happy. years ahead of you. You had a good job, happy family, friends and good. health. No one, not even with the most effective crystal ball, could. have foretold what was to happen. ‘What. you did on that dreadful night was largely outside your control. Through it all you continued to love your mother and she you. After she choked her mother, for a drive in her car wearing only pyjamas and an overcoat. Carol's body was found on her bedroom floor wearing her pyjamas when she failed to turn up for work at Penny Lane Cafe. ‘What. you did was wholly out of character – and you are a loving and caring. person who will find it difficult in the future to live with this. episode.’ Liverpool Crown. Court heard that the killing, in June last year, followed six months of. unhappiness for Miss Parr, who worked as a data manager in the Molecular. and Clinical Cancer Medicine department at the University of Liverpool. She and her mother had been as close as sisters, living together,. socialising and holidaying in exotic locations like Dubai. But. Miss Parr’s life began to crumble following a break-up in January last. year. Simon Berkson, defending, told the court she had been devastated. by the end of her romance at a time when she may have had hopes of. settling down and starting a family. The. court heard Miss Parr became introspective and her relationship with. her mother was ‘strained and tense’. In May, she took an overdose of. paracetamol in a suicide attempt. She became anxious about the damage. she might have done to her liver, and began following her mother around. and sleeping in her bed at night. On. the evening of June 9 last year, Mrs Parr had had enough of her. daughter’s behaviour. She told her she was thinking of selling their. home in Walton, Liverpool, as she couldn’t cope any longer. Emma was seen by neighbours climbing out of a bedroom window onto a flat roof and folding clothes as if to go on a trip. Carol's body was found on her bedroom floor wearing her pyjamas when she failed to turn up for work. The mother and daughter lived, socialised and even holidayed together and had a close relationship 'akin to sisters' A. struggle followed and at 2am Miss Parr dialled 999 to request an. ambulance. The operator could hear ‘groaning noises’ in the background,. but Miss Parr then said paramedics were not needed. Her mother’s body. was found on her bedroom floor later that day after she failed to turn. up at the cafe where she worked as a waitress. Officers discovered Miss Parr several hours later, driving her car in her pyjamas with bare feet in Runcorn, Cheshire. She. was detained at a psychiatric clinic for nearly six months under the. Mental Health Act before being charged with her mother’s murder in. November. Mrs Parr’s. brother Thomas Gregory told the court the family had been ‘torn apart’ by the tragedy but he loved his niece and was supporting her. ‘We can’t. turn back the clock and must face the future. Hopefully, with support. from all of us Emma will be able to have a normal life. Carol would not. want Emma to suffer in any way.’ Miss Parr’s brother, Jason, a father of two, added: ‘Justice is needed for my mum but I need my sister and my sister needs me.’

Summary: Emma Parr, 38, turned on her mother, Carol, 62, after she became depressed following a break up. Attack happened when Carol revealed she planned to sell the family home. Emma recollected little of what had happened only recalling her 'disappointment' at learning her mother no longer wanted to live with her. She admitted manslaughter on the grounds of diminished responsibility. Emma was jailed for ten months after a judge ruled she was a 'caring and loving person' whose attack was 'largely outside her control'

cnn_dailymail

en

Summarize: BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates to an improved angled rotating surgical instrument. More particularly, this invention relates to an angled rotating surgical instrument wherein suction or aspiration may be effectively applied to the surgical site. 2. Discussion of the Prior Art U.S. Pat. No. 4,646,738 to Trott, assigned to the assignee of the present application, shows a rotary surgical tool wherein a surgical cutter is mounted for rotation at the end of an angled, elongated rigid sheath. The cutter is driven by a flexible torque-transmitting member rotating within the sheath. A proximal end of the sheath is mounted to a housing of a motor, typically disposed in a handpiece. The motor is coupled to the torque-transmitting member to drive the rotary cutter. The sheath has a bend near its distal tip which allows the surgeon access to several different portions of the surgical site through a single incision. Such bent or angled surgical instruments are becoming increasingly popular. In order that the torque-transmitting member can drive the cutter while extending around the bend in the sheath, the torque-transmitting member must be freely flexible. According to the Trott invention, the torque-transmitting member comprises three concentric flexible tubular spring-like members. Each of the flexible tubular members is a continuous spiral-wrapped stainless steel wire of generally flat cross-section. The central of the three concentric members is wound with the opposite hand with respect to the hand of the inner and outer members, so that upon application of torque, the assembly tends to tighten upon itself. The three concentric members are secured to one another at their ends by spot welding or a like expedient. The disclosure in the Trott patent is expressly incorporated herein in its entirety by this reference. Another generally relevant device is shown in European Patent Application No. 445,981 of Krause et al, the disclosure in which, and in any U.S. counterpart patent, is expressly incorporated herein in its entirety by this reference. In Krause et al, a flexible torque-transmitting member for a surgical instrument having an angled probe is provided by forming a number of opposed rows of interdigitated circumferential slits extending through the wall of the tubular metallic member, or by drilling a number of offset rows of holes therethrough. In order to remove debris from the surgical site, it is desirable to apply suction or aspiration to the proximal end of the instrument, the suction or aspiration being effectively conveyed to the surgical site by way of the lumen of the tubular torque-transmitting member. In the Trott device, the fact that the torque-transmitting member comprises three concentric spiral-wrapped members allows substantial suction or aspiration to be transmitted from the proximal end to the distal tip of the instrument. However, some leakage takes place between the substantially circumferential slits formed between the adjacent spiral-wrapped wire sections, so that improvement in the efficiency of transmission of the suction or aspiration would be desirable. In the Krause et al device, the slits or holes in the torque-transmitting member may be filled with a silicone rubber or other pliable material to permit the necessary flexibility yet allow the suction or aspiration to be effectively conveyed to the distal tip of the instrument. Such pliant plugs tend to fall out of the slits, destroying their beneficial effect. Other generally related subject matter is shown in U.S. Pat. No. 4,858,897 to Irifune showing a spring which might be used as a flexible tension member within a sheath in an instrument of the type shown in the Trott patent. The Irifune spring would also require protection against loss of suction as in the case of the Krause et al European patent. U.S. Pat. No. 4,576,772 to Carpenter shows a fluid-tight catheter having notches cut in its surface to facilitate bending. German OS 3,219,169 of Nagel et al apparently shows various ways in which a tubular member may be perforated or slit to impart flexibility thereto. If the Nagel et al device were used as a torque-transmitting element in a probe, suction or aspiration applied to one end thereof would not be effectively transmitted to the other end. OBJECTS AND SUMMARY OF THE INVENTION It is accordingly an object of the invention to provide an improved angled rotating surgical instrument wherein suction or aspiration applied to a proximal end of the instrument is effectively communicated to the distal end thereof. It is a further object of the invention to reduce friction between the torque-transmitting member and the rigid outer sheath of an angled rotating surgical instrument, to ease assembly and disassembly of the instrument for cleaning, to provide vibration absorption, and to eliminate any necessity for forming a costly, low-friction coating on the inner surface of the sheath. These and other objects of the invention which will appear as the discussion below proceeds are satisfied by the present invention, whereby a thin sleeve of a fluid-impermeable material is placed over the flexible torque-transmitting member rotating within the sheath. Typically, the sleeve is of a so-called heat-shrink polyester material as used for electrical insulation and other such purposes; however, other materials suitable for performing the functions described herein may be employed. The sleeve is placed over the torque-transmitting member and, if made from heat-shrink polymer, is heated, whereupon the sleeve shrinks into place and is retained on the torque-transmitting member. The fluid-impermeable sleeve material has low surface friction and acts as an effective lubricant between the torque-transmitting member and the outer rigid sheath. The presence of the continuous sleeve over the flexible section of the torque-transmitting member effectively seals the torque-transmitting member, such that suction or aspiration applied to the proximal end of the instrument is effectively transmitted to the surgical site. Other objects and advantages of the present invention will become apparent from the following description of the preferred embodiment, taken in conjunction with the accompanying drawings wherein like parts in each of the several figures are identified by the same reference characters. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows a partial broken cut away view of the flexible section of the torque-transmitting member; and FIG. 2 shows an exploded broken partially sectional view of the complete instrument of the invention comprising a handpiece, a torque-transmitting member with a termination, and an outer rigid sheath with a corresponding termination. DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 shows the flexible portion of the torque-transmitting member of the invention, including three concentric members 10, 12 and 14 as disclosed in the Trott patent. Members 10, 12 and 14 comprise spiral-wrapped stainless steel wire. The handedness of the inner and outer members 10 and 14 may be opposite to that of the intermediate member 12. As disclosed in the Trott patent, this arrangement of the concentric members ensures that application of torque to the assembly will cause the spiral wrap members to contract upon one another. To ensure proper operation the three concentric members may be secured to one another at their ends by a spot-weld or the equivalent as indicated generally at 15. According to the present invention, the assembly of the three concentric members 10, 12 and 14 is covered with a very thin sleeve 16, preferably of a polymeric material and typically of a polyester heat shrinkable material. After fabrication of the flexible member comprising the three concentric members 10, 12 and 14, and their spot-welding to one another at 15, the polymeric sleeve 16 is assembled thereover and heat is supplied to shrink the sleeve snugly onto the assembly. In a preferred embodiment the polymeric sleeve is polyester heat shrink tubing having a wall thickness on the order of 0.00073 inches. In other respects, the flexible section 20 of the torque-transmitting member 22 (FIG. 2) is as disclosed in the Trott patent. Each of the three concentric members 10, 12 and 14 may be formed by wrapping a flat stainless steel wire 0.125 inches wide by 0.003 inches thick around a mandrel of appropriate diameter. As shown in FIG. 2, the complete angled rotary surgical instrument according to the invention includes a handpiece 30, a torque-transmitting member indicated generally at 22 and including a flexible section 20 detailed above in connection with FIG. 1, and a rigid tubular sheath 32. As indicated by the dot-dash lines in FIG. 2, in use the flexible torque-transmitting member 22 slides within the rigid sheath 32 such that the rotating cutter assembly 34 is supported at the distal tip 36 of the rigid sheath 32. The cutter assembly 34 ma comprise a stationary element 38 mounted on the sheath 32, and a rotating element 40 mounted on the torque-transmitting member 22 for rotation with respect to the stationary member 38. Preferably, the rigid sheath 32 and the torque-transmitting member 22 are provided with terminal members 42 and 44, respectively, as shown schematically in FIG. 2. The sheath 32 and torque-transmitting member 22 are thus adapted for quick release connection to the handpiece 30. Such connections are generally known to the art and may be implemented in a conventional fashion. When the torque-transmitting member 22 and the sheath 32 have been assembled to the handpiece 30, termination 44 of the torque-transmitting member 22 is coupled to a shaft 46 of a drive motor 48 for rotating the torque-transmitting member 22 and the cutter assembly 34 within sheath 32. At the same time, suction or aspiration may be applied to a lumen extending the length of the torque-transmitting member 22 by way of a port 50 in termination 44 communicating with a passage 52 in the handpiece 30, to which is connected a source 54 of suction or aspiration as desired. According to the invention, the fluid-impermeable sleeve member 16 allows suction or aspiration applied to the proximal end of the torque-transmitting member 22 to be effectively communicated to the distal end thereof. Since the torque-transmitting member comprises a number of substantially circumferential slits formed between the spiral-wrapped wire members, if the distal cutting window becomes clogged or obstructed, the condition known as &#34;blow-by&#34; or unrecoverable loss of suction may occur in the absence of sleeve 16. The instrument of the invention is thus assembled for use by inserting the distal tip of the torque-transmitting member 22 into the terminal member 42 of the rigid sheath 32, and sliding the termination 42 of the sheath into recess 56 of the handpiece 30, such that the termination 44 engages an inner corresponding recess 53 in the handpiece 30, and so that motor shaft 46 engages termination 44. At that point the rotating cutter element 40 fits snugly within the stationary member 38 fixed at the distal end of the ridged sheath 32 and is supported thereby. When the motor 48 is subsequently actuated by motor control 58, the torque-transmitting member rotates, rotating cutter 40 with respect to stationary cutter 38, thereby forming an effective cutting instrument. Suction or aspiration may then be provided at 54 to withdraw or flush debris from the surgical site. As indicated above, the provision of sleeve 16 over the flexible portion 20 of torque-transmitting member 22 provides a number of useful functions. The fluid-impermeable sleeve seals the flexible member 20 despite the presence of essentially circumferential slits formed between the flat wires of the concentric members 10, 12 and 14. Stated differently, sleeve 16 provides a seal over the discontinuous surface of flexible portion 20 of torque-transmitting member 22. Sleeve 16 further provides a low friction interface between the flexible member 20 and the interior of the rigid sheath 32. This not only eases assembly and disassembly of the instrument, but also eliminates any necessity of providing a costly low friction coating on the interior surface of the rigid sheath 32. As noted above, the preferred embodiment of the invention utilizes heat-shrink polyester tubing for sleeve 16. It will be appreciated, however, that other materials may be employed and need not be applied by heat shrinking. It is only necessary that the resulting sleeve be capable of performing the functions ascribed thereto in the foregoing description. The most important of these functions is sealing the torque-transmitting member in order that the distal end of the cutter may be efficiently aspirated from the proximal end. In addition, the sleeve should provide the bearing and vibration damping functions described above, although these are not as critical as fluid impermeability. Of course, the sleeve also must not interfere with the rotation of the torque-transmitting member. One alternative to the heat shrink polyester material is Teflon which has all of the desirable features required for sleeve 16. A Teflon sleeve 16 would take the form of a tube journaled or &#34;free-wheeling&#34; about the torque-transmitting member. Still another alternative embodiment of sleeve 16 is a &#34;spray-on&#34; resin or other material that is sprayed as a liquid and solidifies on the torque-transmitting member. It should also be noted that the fluid-impermeable sleeve of the present invention is useful to preclude aspiration leakage in instruments other than angled cutters. Thus, the sleeve is useful in straight cutters and in non-cutting instruments through which aspiration of a surgical site is effected. It is also useful in cutters or the like that are conformable to shape by the end user. Such a cutter may be supplied by its manufacturer without bends but is bent to a desired angle before use. While as indicated the cooperating cutting elements may be permanently fixed to the distal ends of the tubular sheath and torque-transmitting member, it is also within the scope of the invention to provide separable cutter assemblies. Other aspects of the construction of the complete instrument according to the invention are generally as taught in the Trott patent and need not be reiterated here. However, the invention is not limited to the particular flexible member disclosed by Trott, but is also useful in connection with the device shown in the Krause et al European patent. If the Irifune spring were employed as a flexible element in a rotating surgical instrument it would similarly be substantially improved by placement of a fluid-impermeable sleeve 20 thereover according to the invention. Inasmuch as the present invention is subject to many variations, modifications and changes in detail, it is intended that all subject matter discussed above or shown in the accompanying drawings be interpreted as illustrative only and not be taken in a limiting sense.

Summary: A thin-walled sleeve of polymeric or similar material is heat shrunk or otherwise closely fitted over a flexible torque-transmitting shaft in an angled rotary surgical instrument. The sheath primarily provides a seal to maintain aspiration or suction through the porous flexible shaft, and also provides an additional bearing surface on the distal end of the flexible shaft, facilitates insertion and removal of the flexible shaft, and provides a low friction surface between the flexible shaft and outer sheath of the instrument.

big_patent

en

Write a title and summarize: The innate immune response is essential for controlling West Nile virus (WNV) infection but how this response is propagated and regulates adaptive immunity in vivo are not defined. Herein, we show that IPS-1, the central adaptor protein to RIG-I-like receptor (RLR) signaling, is essential for triggering of innate immunity and for effective development and regulation of adaptive immunity against pathogenic WNV. IPS-1−/− mice exhibited increased susceptibility to WNV infection marked by enhanced viral replication and dissemination with early viral entry into the CNS. Infection of cultured bone-marrow (BM) derived dendritic cells (DCs), macrophages (Macs), and primary cortical neurons showed that the IPS-1-dependent RLR signaling was essential for triggering IFN defenses and controlling virus replication in these key target cells of infection. Intriguingly, infected IPS-1−/− mice displayed uncontrolled inflammation that included elevated systemic type I IFN, proinflammatory cytokine and chemokine responses, increased numbers of inflammatory DCs, enhanced humoral responses marked by complete loss of virus neutralization activity, and increased numbers of virus-specific CD8+ T cells and non-specific immune cell proliferation in the periphery and in the CNS. This uncontrolled inflammatory response was associated with a lack of regulatory T cell expansion that normally occurs during acute WNV infection. Thus, the enhanced inflammatory response in the absence of IPS-1 was coupled with a failure to protect against WNV infection. Our data define an innate/adaptive immune interface mediated through IPS-1-dependent RLR signaling that regulates the quantity, quality, and balance of the immune response to WNV infection. West Nile virus (WNV) is a neurotropic flavivirus and is an emerging public health threat. Infection with WNV now constitutes the leading cause of mosquito-borne and epidemic encephalitis in humans in the United States [1]. WNV is enveloped and contains a single strand positive sense RNA genome of approximately 11 kb in length that encodes three structural (C, prM/M, and E) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5). It cycles enzootically between birds and Culex mosquitoes, with humans infected as dead-end hosts. WNV infection has been modeled in inbred mice wherein infection and pathogenesis recapitulate many of the features of human infection (reviewed in [2]). Following subcutaneous inoculation, WNV replicates in dendritic cells (DCs) at the portal of entry and in the draining lymph node. A primary viremia develops and virus spreads to visceral organs including the spleen, where further amplification occurs, leading to central nervous system (CNS) dissemination and encephalitis. In humans, WNV causes an acute febrile illness that can progress to severe and sometimes lethal neuroinvasive disease, especially in the elderly and immunocompromised [3]. However, healthy young adults are also afflicted with severe neurological disease [4], [5], [6], indicating that virulence can occur independently of immune deficiencies or aging. Intracellular innate immune defenses and the actions of type I interferon (IFN) provide a first-line of defense against virus infection and are essential for the control of WNV replication, dissemination, and neurovirulence [7]. Innate antiviral immune defenses are triggered through the recognition of conserved pathogen associated molecular pattern (PAMP) motifs within viral products by intracellular pathogen recognition receptor (PRR) proteins in infected cells. PRR signaling directs downstream activation of latent transcription factors, including NF-κB, interferon regulatory factor (IRF) -3 and IRF-7, in a cell type-specific manner to induce antiviral response programs that include expression of proinflammatory cytokines, chemokines, type I IFN, and interferon stimulated genes (ISGs) [7], [8], [9], [10]. The ISG products induced through autocrine and paracrine actions of IFN confer antiviral activity by limiting virus replication and cell-to-cell virus spread. Modulation of IFN signaling has been identified as a virulence feature of pathogenic strains of WNV [11], [12]. The RLRs, retinoic acid inducible gene-I (RIG-I) and melanoma differentiation antigen 5 (MDA5) [13], [14], [15], [16], are PRRs that play critical roles in triggering immune defenses against RNA virus infection, including WNV. RIG-I and MDA5 are cytosolic RNA helicases that contain an amino terminal tandem caspase activation and recruitment domain (CARD). Upon engaging RNA substrates, the RLRs undergo a conformational change and bind to the mitochondrial associated protein, interferon promoter stimulator-1 (IPS-1) through a CARD-CARD interaction, leading to IPS-1-dependent signaling of IFN production and expression of immune response genes [17], [18]. RLR signaling and IPS-1 function have an essential role in triggering IFN defenses during WNV infection of mouse embryo fibroblasts (MEFs) and human cell lines in vitro. Cells lacking either RIG-I or MDA5 were attenuated in their ability to generate an effective innate immune response to infection, whereas cells lacking both RIG-I and MDA5 or those deficient in IPS-1 alone were unable to respond to infection with WNV and related flaviviruses [19], [20], [21], [22]. Recent studies examined the role of another class of pattern recognition receptors, Toll like receptor (TLR) 3 and TLR7, and show that these receptors are also important PRRs of WNV infection, as they play a role in signaling IFN production and an inflammatory response upon viral ligand recognition [23], [24], [25]. TLR3 has been shown to contribute to both enhancement and protection of CNS inflammation and neurovirulence of WNV in vivo [23], [24], while TLR7-dependent signaling was shown to be essential for directing proper immune cell homing to sites of WNV infection during the adaptive immune response in vivo [25]. Type I IFN, a major product of PRR signaling, has been shown to link innate and adaptive immune responses. However, the specific PRR pathways that mediate this during acute WNV infection have not been delineated nor has the RLR pathway been evaluated in this context. The quantity and quality of the innate and adaptive immune responses after infection must be carefully regulated to avoid aberrant inflammation and immunopathogenesis. Regulatory T (Treg) cells and inflammatory dendritic cell (DC) subsets regulate inflammation during acute virus infection through T cell suppression and by modulating the trafficking and inflammatory cytokine production of immune cells into infected tissues [26], [27], [28]. Thus, the level of local and peripheral Treg cells, and the composition of local DC subsets that develop during WNV infection may determine immune control and WNV disease. Here, we assessed the role of RLR signaling and IPS-1 in WNV infection and immunity. Our studies define IPS-1 as an essential modulator of immunity in vivo and demonstrate that IPS-1-dependent signaling orchestrates an innate/adaptive immune interface that regulates immune responses to effectively control WNV infection. WNV infection of primary embryonic fibroblasts recovered from RIG-I−/− mice revealed that RIG-I was important in eliciting innate antiviral immune defenses early during infection, whereas MDA5 was important for enhancing and sustaining this response [21]. We further evaluated WNV infection of RIG-I−/− or MDA5−/− mice and confirmed that RIG-I serves a dominant role among the RLRs for the acute induction of innate immune defenses and protection against WNV infection in vivo (data not shown). Since the RLRs signal innate defenses through the IPS-1 adaptor protein [29], we also examined the role of IPS-1 in protection against WNV infection upon a sub-lethal virus challenge of wild type and IPS-1−/− mice. IPS-1−/− mice were highly susceptible to WNV infection and exhibited 100% mortality with an average survival time (AST) of 7. 3 days as compared to wild type mice (38. 5% mortality with an AST of 13. 2 days; p<0. 0001; Fig 1A). Thus, RIG-I and IPS-1-dependent signaling are essential for protection against WNV infection. To define the role of IPS-1 in controlling WNV in vivo, wild type and IPS-1−/− mice were infected subcutaneously (s. c.) with 100 PFU of WN-TX and viral burden within peripheral tissues and the CNS was measured over time post-infection (pi). IPS-1−/− mice exhibited increased viremia compared to wild type mice (45. 7 fold enhancement at day 1 pi, P<0. 05) throughout the course of infection (Fig 1B). Similarly, viral loads in the spleen were elevated in the infected IPS-1−/− mice (Fig 1C). WNV infection of IPS-1−/− mice displayed an expanded tissue tropism as infectious virus was found in the kidneys, a tissue that is not normally permissive to infection in wild type mice (Fig 1D). WNV is typically detected in the CNS of wild type mice after s. c. challenge between 4 and 8 days pi [2]. Consistent with this time course, infected wild type mice exhibited detectable viral loads (average viral titer of 101. 8 pfu/gram of tissue) in the brain by day 6 p. i., although virus was not detected in the spinal cord (Fig 1E and F). In contrast, WNV spread to the brain (Fig 1E) and spinal cord of IPS-1−/− mice (Fig 1F) by day 2 pi, with viral loads rising through day 6 pi. Together these results indicate that IPS-1, likely through RLR signaling of innate immune defenses, limits WNV replication, viremia, and peripheral spread, and is essential for the control of viral invasion of the CNS. Myeloid cells, including tissue and lymphoid DC and macrophages (Mφ), are among the first cells to encounter WNV during infection and thus function to restrict the spread of virus to distant tissues and the CNS [2]. To define the role of IPS-1 in controlling virus replication and innate immunity in myeloid cells, we analyzed WNV infection and host responses in primary bone marrow-derived DC and Mφ recovered from wild type and IPS-1−/− mice. DC and Mφ were infected at an MOI of 1. 0 (relative to viral plaque assay quantification of BHK-21 cells; see Methods) and evaluated for virus replication, IFN induction, and innate immune triggering of ISG expression (Fig 2). IPS-1−/− DCs sustained significantly higher WNV replication at 36 and 48 hours pi compared to wild type infected cells (Fig 2A). WNV infection of wild type DCs induced IFN-β secretion but this response was completely abolished in IPS-1−/− DCs (Fig 2B). The lack of IFN-β induction in IPS-1−/− DCs correlated with a lack of ISG expression including RIG-I, MDA5, and STAT-1 (Fig 2C). In addition, expression of ISG54 and ISG49, which are direct IRF-3 target genes [30], [31], were not induced during WNV infection of IPS-1−/− DCs (Fig 2C). Moreover, ISG56, another IRF-3 target gene [31], was induced late during infection and to lower levels as compared to ISG54 and ISG49 in wild type, infected DCs. WNV infection of IPS-1−/− Mφ resulted in significantly higher virus replication between 24 and 48 hours pi as compared to infected wild type cells (Fig 2D). Whereas wild type infected Mφ expressed IFN-β, this response was completely abolished in IPS-1−/− Mφ (Fig 2F). We also observed a differential expression of ISGs and IRF-3-target genes within WNV-infected Mφ. RIG-I, MDA5, and STAT-1 were not induced in IPS-1−/− Mφ, whereas, ISG56, ISG49, and PKR were expressed at reduced levels and with delayed kinetics. These data establish that IPS-1-dependent RLR signaling is the major innate immune signaling pathway that controls virus replication in conventional DCs and Mφ. Neurons represent the target cell of WNV infection in the CNS and their death after infection is a key factor in pathogenesis and neurological sequelae [32], [33]. To define the role of RLR signaling in restricting virus replication in neurons, primary cortical neurons were generated from wild type and IPS-1−/− mice. Cells were infected at an MOI of 1. 0 with WN-TX and virus yield, IFN-β induction, and ISG expression were evaluated. In the absence of IPS-1, WNV replicated faster and to higher levels resulting in a 2. 2 and 4. 2-fold (p<0. 05) increase in viral production at 24 hrs and 48 pi, respectively as compared to infected wild type neuronal cells (Fig 3A). This relatively modest virologic effect in neurons compared to that observed in IPS-1−/− DC and Mφ was expected, as IFN-α or -β pre-treatment only inhibits WNV infection in cortical neurons to a maximum of 5 to 8-fold [12], suggesting that the IFN response is comparably less potent in neurons. IFN-β expression was induced to lower levels in IPS-1−/− neurons compared to wild type infected neurons at 24 (10-fold, p<0. 05) and 36 hours pi (5-fold, p<0. 05) despite the higher levels of virus replication (Fig 3A and 3B). Expression of ISGs, (including RIG-I and MDA5) and IRF-3 target genes (including ISG56 and ISG49) followed this pattern and were dependent on IPS-1 for rapid and high level expression (Fig 3C). The presence of IFN-β and ISG transcripts in IPS-1−/− cells at 48 hrs pi is consistent with the finding that TLR3 has an independent and subordinate role in triggering innate immune responses in cortical neurons at later time points after WNV infection [23]. These results demonstrate that the RLR signaling pathway controls virus replication and induces innate immune responses against WNV infection in cortical neurons. To determine the role of the RLR pathway in protection of neurons against WNV pathogenesis in vivo, we conducted histological analysis of brain tissue from wild type and IPS-1−/− mice infected with WN-TX (Fig 4A). Analysis of brain sections from infected wild type mice revealed little or no inflammation or neuronal damage, with sparse and focal cell infiltrates restricted to the hippocampus and cerebral cortex on day 6 pi. By day 10 pi (a timepoint in wild type mice in which peak virus replication in the CNS occurs [34]) cellular infiltrates were present in the parenchyma and neuronal destruction was observed throughout the cortex and hippocampus. In contrast, brain sections from infected IPS-1−/− mice recovered on day 6 pi displayed extensive injury to neurons in the cortex and granular neurons of the hippocampus. Damaged neurons appeared pyknotic with vacuolation, degeneration and cell dropout. Somewhat surprisingly, we observed extensive inflammation in the brains from infected IPS-1−/− mice within the cortex, hippocampus, and cerebellum (data not shown) displaying prominent perivascular and parenchymal immune cell infiltrates. To evaluate the composition and antigen-specificity of the inflammatory cells within the brains of WNV-infected mice, lymphocytes were isolated from infected brains on day 6 pi and were characterized from pools (n = 5) of wild type and IPS-1−/− infected mice. Brains from IPS-1−/− infected mice showed an 2. 9-fold increase in the total number of immune cells as compared to wild type infected mice (Fig 4B), and this was associated with an increase in absolute numbers of infiltrating CD4+ and CD8+ T cells (Fig 4C). Among the brain CD8+ T cells isolated from IPS-1−/− mice, there was a remarkable 27-fold increase in the number of antigen-specific cells that expressed IFN-γ after treatment with an immundominant NS4B peptide (Fig 4D) [35], [36]. Analysis of microglia/Mφ cells, based on relative surface expression of CD45 and CD11b [37], revealed increased numbers of microglial cells (CD45+lo/CD11b+) and infiltrating macrophages (CD45+hi/CD11b+) within the brains of infected IPS-1−/− mice when compared to wild type mice (Fig 4E). Similar findings were observed in the spinal cords from infected IPS-1−/− mice (data not shown). Combined with the histological analysis, these results demonstrate that in the absence of IPS-1, WNV infection induces a strong inflammatory response in the CNS. While this response is likely associated with increased viral loads, the failure of this increased inflammatory response to elicit protection or control CNS pathology, in the absence of IPS-1, suggests a role for the RLR signaling pathway as a regulatory program that controls the quality of the inflammatory response to WNV infection. To further characterize how IPS-1 modulates the inflammatory response to WNV infection, we measured levels of systemic type I IFN, proinflammatory cytokines, and chemokines present in the serum of WNV-infected mice at 1 and 4 days pi. Paradoxically, a trend towards more rapid induction and increased levels of type I IFN were observed in the serum of IPS-1−/− mice compared to wild type mice (Fig 5A). We note that in this case type I IFN was detected and quantified through a mouse-specific type I IFN bioassay, which does not differentiate between the IFN-α or -β species. This result is consistent with our recent studies showing that serum type I IFN levels accumulate during WNV infection in an IRF-7-dependent but IRF-3-independent manner [8], [9]. In this case IFN-α species are likely accumulating through a TLR7-dependent signaling pathway involving plasmacytoid DCs, which do not require the RLR pathway for IFN production [38]. More recently, Town et al. assessed the role of TLR7 and MyD88−/− during WNV infection and found that mice lacking MyD88 produced elevated levels of systemic IFN during WNV infection [25]. Thus, during WNV infection systemic IFN is regulated through RLR-dependent and independent processes. Correspondingly, when compared to wild type mice, IPS-1−/− infected animals, which show higher viremia (Fig 1B) produced increased serum levels of proinflammatory cytokines and chemokines in response to WNV infection. Elevated levels of systemic IL-6, TNF-α, CXCL10, and IFN-γ were observed at 1 and/or 4 days pi in IPS-1−/− mice (Fig 5B). Serum cytokine levels were also compared between uninfected wild type and IPS-1−/− mice and showed no differences in basal cytokine expression (data not shown). These results demonstrate that in the absence of IPS-1, greater proinflammatory cytokine and chemokine responses are induced during WNV infection. WNV-specific antibody responses are essential for suppressing viremia and virus dissemination and limiting lethal WNV infection [39], [40]. To determine if a deficiency in IPS-1 modulated the quality and quantity of the humoral immune response, we characterized the antibody profile in sera during WNV infection. In wild type mice, neutralizing virus-specific IgM antibodies are typically detectable by day 4 pi with WNV and production of neutralizing virus-specific IgG antibodies follow between days 6 and 8 pi [40]. A time course analysis in wild type and IPS-1−/− infected mice showed that between 4 and 6 days pi, WNV-infected IPS-1−/− mice exhibited significantly higher levels of virus-specific IgM, IgG, and IgG subclasses as compared to infected wild type mice (Table 1). WNV-specific IgG1 antibodies were detected at low levels on day 6 pi in sera from wild type and IPS-1−/− mice. Additionally, we observed a ∼72. 9-fold increase in WNV-specific IgG2a levels in infected IPS-1−/− as compared to wild type mice on day 6 pi and ∼2. 2-fold increase on day 8 pi. Assessment of the virus-specific antibody responses through a PRNT assay revealed that neutralization titers in sera from wild type mice increased dramatically between 6 and 8 days pi. Sera from IPS-1−/− infected mice exhibited a modest increase in neutralization titer to 1∶1280, despite having much higher levels of virus-specific antibodies. This difference translated into a serum neutralization index that was ∼39-fold lower on day 6 pi in the infected IPS-1−/− mice compared to wild type mice. These results demonstrate that the humoral responses in WNV-infected IPS-1−/− mice are distinct from responses in wild type infected mice. Thus, RLR signaling and IPS-1 actions likely contribute to regulatory processes that govern the levels, IgG class switching, and neutralizing capacity of antibodies generated in response to WNV infection. To characterize the immune parameters associating with the dysregulated inflammatory and humoral responses in WNV infected IPS-1−/− mice, we analyzed the immune cell composition in draining lymph node and spleen tissues. Wild type and IPS-1−/− mice were challenged with diluent alone or with WN-TX, and draining popliteal lymph node (DLN) and the spleen were harvested at 1 and 6 days pi, respectively. Analysis of the popliteal DLN provides insight into how IPS-1 modulates the inflammatory response immediately after infection whereas assessment of the spleen elucidates characteristics of the adaptive immune response prior to the infection endpoint. Immune cells were isolated from the popliteal DLN and were characterized by flow cytometry (Fig 6). Analysis of CD8α DC subsets, which are phenotypically the major antigen presenting cells within lymphoid tissues and are implicated in eliciting virus-specific CD8 T cell in response to acute WNV infection [41], showed that infected wild type and IPS-1−/− mice exhibited similar increases in the numbers of CD8α+ and CD8α− DCs compared to mock-infected mice (Fig 6A, B). However, a significant increase (∼3-fold; p<0. 05) of a proinflammatory DC subset, characterized as CD11c+CD11bhiLy6C+, was observed within the popliteal DLNs of IPS-1−/− infected mice (Fig 6C). This DC subset is monocyte-derived and typically recruited to sites of acute inflammation where they propagate the inflammatory response [42]. We found that these DC subsets were not significantly expanded and showed no differences in their recruitment to the DLN in either wild type or IPS-1−/− infected mice at 12 hours pi (data now shown). Thus, as early as 24 hours pi, an elevated cellular inflammatory response is initiated in the IPS-1−/− mice. In contrast, similar increases in plasmacytoid DCs were observed within infected wild type and IPS-1−/− infected mice (Fig 6D), demonstrating that an absence of IPS-1 did not directly affect expansion and/or recruitment of this DC subset. Within the popliteal DLNs, mock-infected IPS-1−/− mice compared to wild type mice generally showed elevated numbers of B cells, CD4+ T cells (p<0. 05), and CD8+ T cells (Fig 6E, F, and G). These results suggest that IPS-1 contributes to the homeostasis of lymphocyte populations within LNs. WNV infection of wild type mice increased the number of B cells (3. 4 fold), CD4+ T cells (3. 1 fold), and CD8+ T cells (2. 3 fold; p<0. 05) in the DLN within 24 hours pi. Similar increases in B cells were observed upon infection of IPS-1−/− mice. However, the number of CD4+ and CD8+ T cells was reduced in the DLN after WNV infection of IPS-1−/− mice. Thus, in the absence of IPS-1, WNV infection specifically increases the number of inflammatory Ly6c+ DCs but suppresses the overall expansion and/or recruitment of T cells in the DLN. We further analyzed the lymphocyte composition of the spleen on day 6 after WNV infection (Fig 7). Gross pathologic analysis revealed that WNV infection of IPS-1−/− mice results in massive splenomegaly whereas infection of wild type mice induces only a slight increase in spleen size (Fig 7A). Cell analysis revealed increased numbers of total lymphocytes in the spleens of infected IPS-1−/− mice as compared to wild type mice (Fig 7B). Regulatory T (Treg) cells have recently been shown to contribute to the dampening of inflammation and adaptive immune responses during acute virus infections [26], [43], [44]. Moreover, a reduction in the number of circulating Treg in mice leads to enhanced lethality after WNV infection [45]. A time course analysis of Tregs in wild type mice revealed that WNV infection results in a significant increase in the numbers of FoxP3+ T cells as compared to mock-infected mice beginning on day 4 and peaking by day 6 pi (Fig 7C), indicating the expansion of Tregs during acute WNV infection. Despite their marked increase in viral load, the infected IPS-1−/− mice did not display an increase in the numbers of FoxP3+ T cells at any timepoint analyzed. Thus, IPS-1 signaling directly or indirectly impacts Treg proliferation and does so independently of viral load. We also observed that spleens from infected IPS-1−/− mice exhibited significantly increased numbers of CD8+ T cells in comparison to those from infected wild type mice, whereas the expansion of splenic CD4+ T cells in wild type and IPS-1−/− mice were not different (Fig 7D). The spleens from WNV-infected IPS-1−/− mice showed significantly higher numbers (3. 9-fold; p<0. 05) of WNV-specific CD8+ T cells producing IFNγ. To further define the phenotype associated with WNV-induced splenomegaly in IPS-1−/− mice, we also assessed the numbers of NK cells and neutrophils. Spleens from infected IPS-1−/− mice contained greater numbers of NK cells (CD4−CD8−NK1. 1+cells), NKT cells (CD4+/CD8+/NK1. 1+ cells) and neutrophils (CD11b+Gr1+ cells) (Fig 7E). Although WNV-infected wild type mice infected displayed slight increases in the absolute numbers of these specific cell types, a deficiency of IPS-1 resulted in a more marked enhancement of these immune cell populations. In this study, we establish a major role for RLR signaling in protection from WNV pathogenesis, and demonstrate that IPS-1 is critical for the control of WNV infection in vivo. Our studies indicate that IPS-1-dependent RLR signaling functions to establish balanced, effective, and protective innate and adaptive immune responses, and that IPS-1 links adaptive immune regulation with the innate immunity triggered by RLR signaling during WNV infection. In the absence of IPS-1 in vitro, innate immune defense programs of myeloid DCs and macrophages were ablated or severely attenuated. Moreover, in vivo analysis of infected IPS-1−/− mice showed altered IgG and IgM antibody responses with diminished virus neutralization activity. The inflammatory response to WNV infection is regulated by IPS-1-dependent processes such that a deficiency of IPS-1 resulted in elevated proinflammatory cytokine and chemokines and increased numbers of inflammatory DCs, antigen-specific T cells, natural killer cells, and neutrophils in lymphoid organs, and activated macrophage/microglial cells within the CNS. The dysregulated inflammatory response to WNV infection in IPS-1−/− mice was associated with a reduction in the numbers of Treg cells and their failure to expand during acute infection. These observations demonstrate the critical role of IPS-1 in mediating RLR signaling of innate antiviral immunity against WNV infection, and reveal novel features of IPS-1 function in regulating immune homeostasis, inflammation, and adaptive immunity to infection. Although infection of primary DCs, macrophages, and neuronal cells failed to induce type I IFN upon WNV infection, WNV-infected IPS-1−/− mice showed enhanced systemic type I IFN responses. This finding agrees with previous studies that indicate both IPS-1-dependent and -independent pathways contribute to the systemic type I IFN production in vivo [8], [9], [23], [25]. Most importantly, the enhanced tissue tropism and rapid viral entry into the CNS observed in the IPS-1−/− mice is not affected by the elevated systemic IFN responses. This suggests that local tissue-specific and intracellular responses triggered by RLR-dependent signaling are more essential for reducing viral burden and dissemination. One possible explanation is that a distinct set of RLR-responsive genes function to control virus replication at the site of infection. This could explain, in part, the elevated levels of virus replication, enhanced tissue tropism and cell-to-cell spread in mice or cells deficient in IRF3 or IRF-7, each of which are downstream transcription factors of RLR signaling [8], [9], [10]. Additionally, it is likely that pDCs, which are specialized dendritic cells for producing systemic type I IFN during a viral infection [46], are likely contributing to the IFN responses observed during WNV infection. Studies by Silva et al. have shown that WNV triggers IFN induction in pDCs through a replication-independent manner [47]. Interestingly, within the DLN, we observed similar expansion of pDCs between wild type and IPS-1−/− infected mice, yet at the same timepoint (24 hours pi), elevated systemic type I IFN responses were observed in IPS-1−/− mice. This suggests two possibilities: 1) splenic pDCs or circulating pDCs are likely responding to the high levels virus in the serum from the IPS-1−/− infected mice to produce IFN at 24 hours pi and/or 2) various other cell types that express TLR3 and/or TLR7 are responding to WNV infection and contributing to systemic IFN responses. Taken together, these studies indicate that RLR signaling and the actions of IRF-3/7 are important in triggering IFN production and ISG expression to limit WNV replication and spread, and that TLR signaling may impart additional, RLR-independent defenses that regulate immunity against WNV infection. The production of and response to type-I IFN is a major linkage point between innate and adaptive immunity, as IFN-α and IFN-β sustain B cell activation and differentiation [48], [49], [50], expand antigen-specific CD8+ T cells [51], [52], CD4+ T cells [53], and activation of NK cells [54]. One of the most intriguing aspects of this study was the global alteration of the immune response elicited in the IPS-1−/− mice, indicating that RLR signaling couples innate immunity with regulation of the adaptive immune response. Infection of IPS-1−/− mice exhibited increased IgM and IgG WNV-specific antibodies, enhanced WNV-specific CD8+T cell response, and increased expansion of neutrophils, NK cells and NK-T cells. One trivial explanation for these differences is that there is an increased antigen load in the absence of IPS-1 and, as a result, enhanced virus-specific (e. g. CD8+ T cells, IgG and IgM antibodies) and nonspecific (e. g. Neutrophils, NK cells) responses. However, there are several key findings from this study that argue against these responses simply being attributed to higher antigen load: (1) In the absence of IPS-1, the CD4 and CD8 T cells, which are protective against WNV infection [34], [35], [36], [55], [56], [57], [58], were significantly enhanced in the peripheral and CNS compartments but failed protect against infection. One explanation for this observation is that the expansion and migration of CD4 and CD8 T cells to different tissues was itself uncontrolled, resulting in T cell-mediated pathology rather than T cell-mediated protection. (2) While the quantity of virus-specific IgM and IgG antibody responses were greatly enhanced in the absence of IPS-1, there was a marked reduction in antibody quality in terms of neutralization capacity. In contrast deficiencies in TLR3 or MyD88 (data not shown) did not alter virus-specific antibody responses and neutralization capacities. Collectively, these findings suggest that RLR-dependent signaling coordinates effective antibody responses during WNV infection through as yet undefined pathway. (3) While systemic IFN responses provide a link between innate and adaptive immune responses, our studies suggest that the PRR signaling pathways (RLR-dependent vs –independent) and levels of IFN production in combination with production other proinflammatory cytokines or chemokines regulate the quantity and quality of the immune response during virus infection. Thus, in the absence of IPS-1 signaling, infected conventional DCs or Mφ, two integral cell types in establishing adaptive immunity, likely do not produce IFN or the normal array and level of proinflammatory cytokines/ chemokines. Instead, IFN and other mediators may be strictly produced by infected or bystander cells during WNV infection, occurring with altered kinetics and magnitude, through TLR-dependent pathways, such as TLR3 and/or TLR7 [23], [25]. (4) In the absence of IPS-1, the enhanced expansion of Ly6C+ “inflammatory” DCs failed to limit early WNV replication and dissemination. This inflammatory DC subset migrates to sites of infection, secretes pro-inflammatory cytokines, and promotes CD8+ T cell expansion during a secondary virus infection, suggesting it sustains the effector T cell response [59]. Our data indicate that Ly6C+ DC recruitment and/or expansion is governed by IPS-1-dependent events of RLR signaling. Thus, the aberrant recruitment/expansion of these inflammatory DCs may contribute to immunopathogenesis and limit development of an effective immune response to control WNV virus infection. (5) The lack of Treg expansion during WNV infection correlated with altered IFN levels, increased proinflammatory cytokines and chemokine levels, and an increased number and distribution of antigen-specific CD8+ T cells. These observations implicate an indirect or direct role for IPS-1 in regulating Treg levels during WNV infection, and provide evidence that links a lack of Treg expansion to immune dysregulation. While their importance in autoimmunity is established [60], recent studies have implicated an integral role for Tregs in controlling inflammation and adaptive immune responses during acute virus infections [26], [43], [44]. During acute infection Tregs function to locally contain and control the immune response with the dual outcome of suppressing viral dissemination while decreasing the likelihood of immune-mediated pathology. In support of this model, infection studies with herpes simplex virus (HSV) and mouse hepatitis virus (MHV), two well established models of viral encephalitis, have demonstrated the importance of Tregs in limiting proinflammatory cytokine and chemokine responses to protect the CNS and enhance survival [26], [43]. Recent work also implicates Tregs in the control of WNV pathogenesis, wherein peripheral expansion of Tregs was associated with asymptomatic infection among WNV-infected blood donors but reduced Treg levels associated with WNV disease [45]. Furthermore, these studies revealed that the conditional depletion of Treg cells in mice results in enhancement of WNV virulence and expansion of antigen-specific CD8 T cells. Interestingly, from our studies, type I IFN does not appear to be the major contributor to Treg expansion during WNV infection, as Tregs failed to expand in the IPS-1−/− infected mice despite their elevated levels of systemic type IFN. These observations suggest that RLR signaling through IPS-1 provides essential signals that directly or indirectly impart the expansion of Tregs during WNV infection. We propose that IPS-1 coordinates an innate/adaptive immune interface wherein IPS-1- signaling after RLR engagement regulates the quantity, quality, and balance of the subsequent immune response. The integrity of the innate/adaptive immune interface is central to the eliminating virus but also restricting immunopathogenesis and inflammation during infection. RLR signaling is essential for triggering the innate immune response to RNA viruses that cause human disease, including the influenza viruses, respiratory syncytial virus and other paramyxoviruses, picornaviruses, reoviruses, flaviviruses, and hepatitis C virus [14], [19], [22]. Thus, in addition to WNV, IPS-1-dependent RLR signaling will likely have a broad impact for the control of inflammation, immune response quality, and viral disease. BHK21 and L929 cells were maintained in Dulbecco' s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2mM L-glutamine, 1 mM sodium pyruvate, antibiotic-antimycotic solution, and 1× nonessential amino acids (complete DMEM). WNV strain TX 2002-HC (WN-TX) was isolated by as previously described [11]. Working stocks of WN-TX were generated by a single round of amplification on Vero-E6 (ccl-81; ATCC) cells, and supernatants were collected, aliquoted, and stored at −80°C. Virus stocks were titered by a standard plaque assay on BHK21 cells as previously described [40]. IPS-1−/− (C57BL/6×129Sv/Ev) and their wild type littermate control mice have been published [38], [61] and were obtained as a generous gift from Dr. S. Akira (Osaka University, Osaka, Japan). Mice were genotyped and bred under pathogen-free conditions in the animal facility at the University of Washington. Experiments were performed with approval from the University of Washington Institutional Animal Care and Use Committee. The methods for mice use and care were performed in accordance with the University of Washington Institutional Animal Care and Use Committee guidelines. Age-matched six to twelve week old mice were inoculated subcutaneously (s. c.) in the left rear footpad with 100 PFU of WN-TX in a 10 µl inoculum diluted in Hanks balanced salt solution (HBSS) supplemented with 1% heat-inactivated FBS. Mice were monitored daily for morbidity and mortality. For in vivo virus replication studies, infected mice were euthanized, bled, and perfused with 20 ml of phosphate-buffered saline (PBS). Whole brain, spinal cord, kidney, and spleen were removed, weighed, homogenized in 500ul of PBS, and titered by plaque assay. Bone-marrow derived DC and Mφ were generated as described previously [9]. Briefly, bone marrow cells from wild type and congenic deficient mice were isolated and cultured for 7 days in either RPMI-1640 supplemented with granulocyte-macrophage-colony stimulating factor, and interleukin-4 (Peprotech) to generate myeloid DC or in DMEM supplemented with macrophage colony stimulating factor (Peprotech) to generate Mφ. On day 7, DC or Mφ were infected with WN-TX at an MOI of 1. 0 and at 12,24,36, and 48 hours post-infection (hpi), supernatants were collected for titration of viral burden by plaque assay on BHK21 cells and levels of IFN-β (described below). Cells were collected in parallel for western blot analysis. Cortical neurons were isolated from 15-day-old embryonic mice and cultured as described previously [62]. On day 6 of culture, neurons were infected with WN-TX at an MOI of 1. 0 and at 12,24,36, and 48 hpi, supernatants were collected for virus titration by plaque assay on BHK21 cells and cells were collected for RNA analysis by RT-qPCR (described below). Cells were lysed in modified RIPA buffer (10mM Tris [pH 7. 5], 150mM NaCl, 0. 5% sodium deoxycholate, and 1% Triton X-100) supplemented with protease inhibitor cocktail (Sigma) and phosphatase inhibitor cocktail II (Calbiochem). Protein extracts (25 µg) were analyzed by immunoblotting as described previously [11]. The following primary antibodies were used to probe blots: mouse anti-WNV from the Center for Disease Control; rabbit anti-ISG56, rabbit anti-ISG54, rabbit anti-ISG49, kindly provided by Dr. G. Sen; mouse anti-PKR from Santa Cruz; rabbit anti-RIG-I and rabbit anti-MDA5 from IBL; mouse anti-tubulin from Sigma; and rabbit anti-STAT-1 from Cell signaling. Secondary antibodies included peroxidase-conjugated goat anti-rabbit, goat anti-mouse, donkey anti-rabbit, and donkey anti-mouse were from Jackson Immunoresearch. For analysis of viremia, serum was separated (BD Microtainer tube SST) and RNA was extracted as previously described [8]. WNV RNA copy number was measured by RT-quantitative PCR (RT-qPCR) as previously described [63]. For cultured cells, total RNA was extracted using the RNeasy kit (Qiagen), DNase treated (Ambion) and evaluated for ISG49, ISG56, IFN-β, RIG-I, and MDA5 RNA expression by one-step SYBR Green RT-qPCR. Specific primer sets for ISG-49, ISG-56, RIG-I, and IFN-β have been described previously [30], [64]. Primer sets for MDA5 are: 5′-GTGGTCGAGCCAGAGCTGAT and 3′- TGTCTCATGTTCGATAACTCCTGAA. IFN-α and -β were measured in sera using a biological assay as previously described [65]. Briefly, L929 cells were seeded at 3×104 cells/well in a 96 well plate one day prior to the addition of interferon standards or experimental samples. Mouse sera (diluted 1∶10 in L929 media) were treated with UV light for 20 minutes to eliminate residual virus. Duplicate sera samples were then added to the 96-well plates in two-fold dilutions along with a murine IFN-β standard. The following day, EMCV challenge virus was added to the cells in 50 µl/well at an MOI of 5. 0. Twenty-four hours later, cytopathic effect was measured by a blinded scorer and IFN levels in the sera was calculated based on the IFN standard. IFN-β in cell culture supernatants was analyzed using mouse-specific ELISA kits from PBL Biomedical Laboratories according to the manufacturer' s protocol. WNV-specific IgM, total IgG, IgG1, and IgG2a levels were determined by an ELISA using purified recombinant E protein as previously described [55]. The neutralization titer of serum antibody was determined by using a previously described plaque reduction neutralization assay [40]. Briefly, sera samples from mock or WN-TX infected mice were diluted in DMEM followed by incubation at 56°C for 30 minutes to inactivate virus and complement factors. Sera were further diluted in two-fold increments and incubated with 100 PFU of WN-TX at 37°C for 1 hour. Standard plaque assays were performed on BHK21 cells and the dilution at which 50% of plaques were neutralized was determined by comparing the number of plaques formed from WNV-infected sera samples to mock infected sera samples. WNV infected sera were analyzed for the presence and levels of TNF-α, IFN-γ, CXCL10 (IP-10), and IL-6 by a mouse-specific cytokine/chemokine Milliplex ELISA (Millipore). Mock-infected or WNV-infected mice were exsanguinated and perfused with PBS, 4% paraformaldehyde, pH 7. 3. Brains were embedded in paraffin and 10-µm sections were prepared and stained with hematoxylin and eosin (H&E) by the UW histology pathology laboratory. Sections were analyzed using a Nikon Eclipse E600 microscope (UW Keck microscope facility). Draining lymph nodes from mice were isolated and digested with collagenase (Roche) and type I DNase in serum-free RPMI media at 37°C for 40 minutes with mechanical disruption. Cells were then incubated with RPMI media containing 10% FBS with EDTA and HEPES for 10 minutes at room temperature, pelleted, and resuspended in PBS containing 2% FBS and 0. 1% sodium azide (FACS Staining buffer). Splenocytes were isolated, washed, and re-suspended in RPMI 1640 containing 10% FBS before in vitro stimulation. Cells were washed twice before FACS staining. For isolation of CNS immune cells, mice were euthanized and perfused extensively with PBS to remove residual intravascular leukocytes. Brains and spinal cords from 5 mice per experimental group were isolated and pooled. Tissues were minced in RPMI media, triturated, and digested with Liberase (Roche) and type I DNase in serum-free RPMI media at 37°C for 45 min. Immune cells were isolated after gradient centrifugation from a 37/70% Percoll interface and washed twice with FACS staining buffer. Immune cells were stained with antibodies specific to CD11c, CD11b, B220, CD3, CD25, CD4, CD8, NK1. 1, Gr-1, siglec H, and CD45 (all reagents from eBiosciences). Intracellular FoxP3 staining was performed as previously described [26]. Intracellular IFN-γ staining was performed on splenocytes and CNS immune cells as previous described [35], [36]. Briefly, lymphocytes were stimulated with 1 µg/ml of the WNV NS4B peptide (SSVWNATTAI) for 4 h at 37°C. Cells were washed and stained for cell surface markers followed by permeabilization-fixation using the Cytofix-Cytoperm Kit (BD-PharMingen) and stained with a Pacific-Blue conjugated IFN-γ antibody (eBiosciences) at 4°C for 30 min, washed and analyzed by flow cytometry. Flow cytometry was performed on a BD LSRII machine using BD FACSDiva software. Cell analysis was performed on FlowJo (v. 8. 7. 2) software. For in vitro studies and immune cell analysis an unpaired student T-test was used to determine statistical differences. For in vivo viral burden analysis, Mann-Whitney analysis was used to determine statistical differences. Kaplan-Meier survival curves were analyzed by the log-rank test. A p-value≤0. 05 was considered significant. All data were analyzed using Prism software (GraphPad Prism5).

Title: IPS-1 Is Essential for the Control of West Nile Virus Infection and Immunity Summary: West Nile virus (WNV) is a mosquito-transmitted RNA virus that has emerged in the Western hemisphere and is now the leading cause of arboviral encephalitis in the United States. However, the virus/host interface that controls WNV pathogenesis is not well understood. Previous studies have established that the innate immune response and interferon (IFN) defenses are essential for controlling virus replication and dissemination. In this study, we assessed the importance of the RIG-I like receptor (RLR) signaling pathway in WNV pathogenesis through analysis of mice lacking IPS-1, the central adaptor molecule of RLR signaling. Our studies revealed that IPS-1 is essential for protection against WNV infection and that it regulates processes that control virus replication and triggering of innate immune defenses. We found that IPS-1 plays an important role in establishing adaptive immunity through an innate/adaptive interface that elicits effective antibody responses and controls the expansion of regulatory T cells. Thus, RLRs are essential for pathogen recognition of WNV infection and their signaling programs help orchestrate immune response maturation, regulation of inflammation, and immune homeostasis that define the outcome of WNV infection.

lay_plos

en

Write a title and summarize: Symptomatic acute schistosomiasis mansoni is a systemic hypersensitivity reaction against the migrating schistosomula and mature eggs after a primary infection. The mechanisms involved in the pathogenesis of acute schistosomiasis are not fully elucidated. Osteopontin has been implicated in granulomatous reactions and in acute hepatic injury. Our aims were to evaluate if osteopontin plays a role in acute Schistosoma mansoni infection in both human and experimentally infected mice and if circulating OPN levels could be a novel biomarker of this infection. Serum/plasma osteopontin levels were measured by ELISA in patients with acute (n = 28), hepatointestinal (n = 26), hepatosplenic (n = 39) schistosomiasis and in uninfected controls (n = 21). Liver osteopontin was assessed by immunohistochemistry in needle biopsies of 5 patients. Sera and hepatic osteopontin were quantified in the murine model of schistosomiasis mansoni during acute (7 and 8 weeks post infection, n = 10) and chronic (30 weeks post infection, n = 8) phase. Circulating osteopontin levels are increased in patients with acute schistosomiasis (p = 0. 0001). The highest levels of OPN were observed during the peak of clinical symptoms (7–11 weeks post infection), returning to baseline level once the granulomas were modulated (>12 weeks post infection). The plasma levels in acute schistosomiasis were even higher than in hepatosplenic patients. The murine model mirrored the human disease. Macrophages were the major source of OPN in human and murine acute schistosomiasis, while the ductular reaction maintains OPN production in hepatosplenic disease. Soluble egg antigens from S. mansoni induced OPN expression in primary human kupffer cells. S. mansoni egg antigens induce the production of OPN by macrophages in the necrotic-exudative granulomas characteristic of acute schistosomiasis mansoni. Circulating OPN levels are upregulated in human and murine acute schistosomiasis and could be a non-invasive biomarker of this form of disease. Schistosomiasis is a severe tropical disease caused by Schistosoma spp. flatworms that affects over 200 million of people from 76 countries and territories [1]. S. mansoni is the only species in the Americas and Brazil holds the majority of infected individuals with 25 million living in endemic areas and 4–6 million infected [2]. Infected individuals have various clinical manifestations that generally cluster into three distinct forms of the disease: acute, hepatointestinal and hepatosplenic schistosomiasis [2–5]. In patients from endemic areas, the acute phase of schistosomiasis is rarely symptomatic (0. 3%) due to infection early in life (3–4 years-old) and exposure to schistosoma antigens/antibodies against antigens in-utero and/or in breast milk [3]. The majority of chronically infected patients from endemic areas (90–96%) develop the hepatointestinal form of the disease, which is asymtomatic or oligosymptomatic in most cases and characterized by granulomatous inflammation in the liver and intestines, little or no hepatosplenomegaly, and minimal liver fibrosis without any sign of portal hypertension [2,4–7]. A small proportion (4–10%) of infected individuals from endemic areas develops the hepatosplenic form of disease characterized by hepatosplenomegaly, severe liver fibrosis and portal hypertension [2,4–7]. Among individuals from non-endemic areas, the acute form of schistosomiasis mansoni is a systemic hypersensitivity reaction against the migrating schistosomula (pre-postural phase of infection) and mature eggs (post-postural phase of infection). This typically develops within 16–90 days after a primary infection [2]. The burden of infection (and probably host genetic background) dictates the severity of the clinical manifestations: more worm couples produce more eggs and consequently, trigger an exacerbated host immune response [2,8]. The pre-postural phase occurs during the initial 35 days after infection and is caused by immune modifications induced by the schistosomules, immature and adult worms before laying eggs [2]. Cercarial dermatitis may occur soon after infection, but symptoms are more evident when schistosomules/immature worms arrive/grow/mature in the hepatoportal veins (peak 15–21 days post infection) [2]. High fever (38–39°C), cough, abdominal pain, discrete hepatosplenomegaly and nonspecific symptoms such as muscular pain, arthralgia and headache, are observed [2,9]. Blood eosinophilia (10–75% of eosinophils) is frequent [2,9]. Liver biopsy reveals discrete inflammatory infiltrate consisting of lymphocytes, eosinophils, neutrophils and macrophages surrounding schistosomules/immature worms, non-specific portal hepatitis, and sparse focal intralobular necrosis [2]. During this phase a Th1 response is predominant and an increase in pro-inflammatory cytokines such as IL-2, gamma Interferon and TNF alpha is frequently observed [2,3]. Other less frequent clinical manifestations may be present: transverse myelitis or pseudotumoral lesions in encephalon (neural schistosomiasis) [2,9]. The post-postural phase is initiated by egg laying (approximately 35 days post-infection) and egg maturation (which begins about 6 days later) [2]. Symptoms are aggravated, episodes of diarrhea increase, and the patient experience severe weight loss [2,4, 8]. Clinical symptoms can continue until 90 days after infection [2,4, 8]. Severe, toxemic forms of acute disease in which there is massive dissemination of eggs throughout the intestines and lungs may be fatal [2]. Moderate to mild disease spontaneously resolves two to three months after infection [2]. During acute schistosomiasis intense miliary distribution of eggs occurs in the liver, colons, small intestines, visceral peritoneum, abdominal lymph nodes, pancreas and lungs [2]. Periovular granulomas localize on the serosal surface of affected organs and macroscopically appear as translucent granule or nodules [2]. Microscopically, the granulomas are large (over 100 times the size of the egg), necrotic-exudative, and enriched with eosinophils [2], due to the naïve hosts’ hyperergic reaction to novel parasite antigens. In the liver, granulomas are irregularly distributed through the parenchyma and portal tracts and non-specific inflammatory cells frequently surround portal tracts. Because hepatocellular lesions are relatively mild (loss of basophilia, hydropic degeneration and rare focal necrosis), the serum aminotransferases are usually normal or slightly elevated [2,9]. An important feature of acute schistosomiasisis is that all the granulomas are uniformly in the same necrotic-exudative phase of formation, with prominent central necrosis [2]. This finding in liver biopsies is pathognomonic of acute infection. With egg-laying the Th2 immune response starts to suppress the initial Th1 response and IL4, IL5, IL10 and IL13 are the most predominant cytokines [2,3]. The hyperergic, massive granulomas are modulated as the infection evolves to the chronic phase. By around 90 days post-infection, liver granulomas are smaller [2,3, 10–12] and progressively heal by fibrosis [2,7, 10,12]. The symptoms usually disappear due to the modulation of the immune response to the eggs [2]. Because the signs and symptoms of acute schistosomiasis are nonspecific and diagnosis is established by presence of eggs in stools that occurs only six weeks after infection, acute schistosomiasis mansoni is frequently misdiagnosed, under diagnosed or has delayed diagnosis [2,9]. Efforts to develop tests for earlier diagnosis of the disease have been challenging. Unfortunately, lesions similar to those observed in pre-postural phase of human acute schistosomiasis are not observed in mouse models of schistosomiasis mansoni [11,13,14], likely because the granulomas that form in mice are generally less necrotic than those that occur in acutely infected humans [14]. Osteopontin (OPN), a pro-inflammatory cytokine and pro-fibrogenic molecule [15–17], was recently associated with hepatosplenic schistosomiasis mansoni [18]. Soluble egg antigens (SEA) directly induce liver cells to produce OPN. Moreover, serum and hepatic osteopontin levels correlate with the degree of liver fibrosis and the level of portal hypertension, suggesting that this molecule could be a novel biomarker for hepatosplenic schistosomiais mansoni [18]. The authors observed that macrophages, stellate cells and bile ductular cells in/around the granulomatous reaction are the major sources of OPN in schistosomiasis [18]. Osteopontin was also demonstrated to play a role in recruitment and activation of macrophages/Kupffer cells, neutrophils and lymphocytes [15–17,19]. OPN-/- mice injected with S. mansoni eggs develop abnormal granuloma formation in the lung due to reduced macrophage accumulation [20]. Since in acute schistosomiasis the liver is enriched with necrotic-exudative granulomas and there is an exacerbated immune response, our aims were to evaluate if OPN increases in acute Schistosoma mansoni infection of both humans and mice, and to determine if circulating OPN levels might be a novel biomarker of this infection. This was a comparative cross-sectional study. A total of 28 patients with acute schistosomiasis mansoni diagnosed at Tropical Diseases Outpatient Clinic of the University Hospital of Universidade Federal de Minas Gerais (Belo Horizonte, Brazil) from January 2014 to December 2015 were included in the study. Serum samples from acute patients (n = 28; age 19. 8±11. 8 years; 21 males/7 females) and uninfected controls (n = 21; age 27. 86±9. 45 years; 14 males/7 females) were collected for analysis. Formalin-fixed, paraffin-embedded liver needle biopsies were available in a subgroup of patients (n = 5). Plasma samples from uninfected controls (n = 21) and from patients with Hepatointestinal (n = 27; age 35. 66±12. 09 years; 16 males/7 females), Hepatosplenic (n = 39; age 38. 25±9. 4 years; 30 males/9 females) and Acute (n = 3; age 39±25. 23; 3 males) schistosomiasis were also included in the analysis. Diagnosis of acute schistosomiasis was based on epidemiological data (recent contact with stream water in an endemic area), clinical data (cercarial dermatitis, acute enterocolitis, fever, cough, malaise, paraplegia, pulmonary involvement, hepatomegaly and or splenomegaly), laboratory assays (eosinophilia, IgG antibodies against SWAP, S. mansoni eggs in stools or rectal biopsy fragments), and imaging techniques (Ultrasound to observe liver, spleen and intra abdominal lymph node enlargement; MRI to demonstrate spinal cord injury). To be considered as having acute schistosomiasis in the present study the participants had to have more than 1 or more symptoms/signs described above, evidence of infection (parasitologic or serologic) and reported contact with contaminated waters. All patients included in the study were residents of the metropolitan region of Belo Horizonte (capital of Minas Gerais state), a non-endemic area for schistosomiasis mansoni. No previous history of contact with S. mansoni was reported by the patients or parents/guardians. The present study was conducted in accordance with the Declaration of Helsinki (2013) of the World Medical Association and was approved by the Ethics Committee of Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil (UFMG) (Protocol ETIC 204/06). Written informed consent was obtained from all participating subjects or their parents/guardians (on behalf of child participant). All data regarding human participants was anonymized. Female Swiss Webster outbred mice were infected with 50 cercariae of S. mansoni (Feira de Santana strain, CPqGM/FIOCRUZ) for 6,7, 8 weeks (acute phase, n = 15) and 30 weeks (chronic phase, severe fibrosis, n = 8). Uninfected, age- and strain-matched animals were used as controls (n = 8). Liver tissue and serum were collected for analysis. The present study protocol meet the regulation and guidelines of Brazil’s National Animal Experimentation Control Board (CONCEA) and was approved (Protocol 003/2010) by the Ethical Committee for Animal Research of Centro de Pesquisas Gonçalo Moniz, Oswaldo Cruz Foundation, Salvador, Bahia, Brazil (CPqGM/FIOCRUZ). OPN was quantified in the serum (humans and mice) or plasma (humans) using OPN Quantikine ELISA kit (R&D Systems) according to the manufacturer’s protocol. Liver sections were stained with H&E (haematoxylin and eosin) for general histology. Immunohistochemistry (IHC) analysis was performed to evaluate the expression of osteopontin (R&D Systems; Antigen retrieval: 3% pepsin digestion for 10 min at 37°C; 5ug/mL of primary antibody, incubation overnight at 4°C). To confirm that Macrophages produce osteopontin, double IHC was performed using the chromagen DAB (3,3_-diaminobenzidine) for OPN and the chromagen Vina Green for CD68 (a macrophage marker). OPN staining was quantified in 15 x200 fields/sample by computer-assisted morphometry using MetaMorph (Universal Imaging Corp.). OPN (+) bile ducts were counted in 15 x200 fields/sample by three independent observers. The SEA was prepared at Centers for Disease Control and Prevention (CDC) as previously described [21]. The amount of Gram-negative bacterial endotoxin present in the SEA preparation was quantified using the end-point chromogenic limulus amebocyte lysate assay (Lonza). To investigate if macrophages produce osteopontin, primary human Kupffer cells (from Thermo Fisher Scientific) were incubated with 10 μg/ml SEA or 0. 0001 μg/ml LPS (lipopolysaccharide; control, same amount of endotoxin present in the SEA preparation) for 3,6, 12 and 24 hours. RNA was collected for analysis. RNA was extracted using RNeasy mini kit (Qiagen) according to the manufacture’s protocol. Reverse transcription was performed using the First Strand Superscript III kit (Life Technologies) using the random hexamers protocol. Osteopontin mRNA expression was evaluated by real-time PCR (Taqman, Thermo Fisher Scientific). Each sample was analysed in duplicate and target gene levels in treated cells are shown as a ratio to levels detected in corresponding control samples, according to the ΔCT method, relative to the housekeeping gene (18s). The probes were designed by Thermo Fisher Scientific. Results are expressed as means ± S. E. M. (Standard Error of the Mean; for normal distribution variables) or as medians (for non-normal distribution variables). Comparisons between groups were performed using the oneway ANOVA and Student’s t test (parametric) or Kruskal–Wallis one-way ANOVA and Mann–Whitney U test (non-parametric). Significance was accepted at the 0. 05 level; Bonferroni correction was applied when comparing more than two groups. Receiver operating characteristics (ROC) curve analysis was used to investigate if sera OPN levels could be a good biomarker for symptomatic acute schistosomiasis. All statistical analyses were performed using SPSS Statistics 22 (IBM) and Prism 6 (GraphPad). Patients with acute schistosomiasis have increased circulating levels of osteopontin in the plasma (p = 0. 0005 vs non-infected; p = 0. 0005 vs HI and p = 0. 0012 vs HS) (Fig 1A) and serum (p = 0. 0001) (Fig 1B). The plasma OPN levels in acute schistosomiasis are even higher than in patients with hepatosplenic form of the disease (p = 0. 0012) (Fig 1A). We observe that OPN starts to increase in the beginning of the post-postural phase (5–6 weeks post-infection, p = 0. 0005 vs non-infected) and OPN levels peaked 7–11 weeks post-infection (p = 0. 0001 vs uninfected; p = 0. 04 vc 5–6 weeks; p = 0. 001 vs 12 weeks and p = 0. 0001 vs 24 weeks), when the livers are enriched with necrotic-exudative granulomas (Fig 1C). Twelve weeks after infection the symptoms start to disappear, the granulomas reach a modulated state and circulating OPN levels start to fall, reaching levels comparable to uninfected individuals 24 weeks post-infection (Fig 1C). Receiver operating characteristics (ROC) curve analysis demonstrated that serum OPN measurement could be a good biomarker to identify patients with symptomatic acute schistosomiasis mansoni (Area under the curve = 0. 9959; p<0. 0001; 95% confidence interval 0. 9848–1. 007; S1 Fig). In our study population OPN serum test >23. 34 can detect a symptomatic acute patient with 95. 65% sensitivity and 95. 24% specificity (Likelihood ratio = 20. 09). Immunohistochemistry demonstrated that the inflammatory cells in the necrotic-exudative liver granulomas express OPN, especially in the macrophage (epithelioid cells) enriched area around the egg and central necrosis (Fig 1D and S2 Fig). Similar to humans, mice in the acute phase of infection also have more circulating and hepatic OPN levels than mice in the chronic phase of infection where there is severe fibrosis (p = 0. 001 vs Non-infected; p = 0. 0124 vs chronic phase) (Fig 2A, 2C and 2D). OPN levels in mice also peaked in the liver (p = 0. 0001 vs non-infected; p = 0. 0245 vs 6 weeks and p = 0. 0104 vs 30 weeks) and serum (p = 0. 0286 vs non-infected; p = 0. 0286 vs 6 weeks; p = 0. 0286 vs 8 weeks and p = 0. 004 vs 30 weeks) 7 weeks post-infection, at a time when the livers were enriched with necrotic-exudative granulomas and inflammatory cells (Fig 2B, 2C and 2D). During the acute phase of infection in both mice and humans, the majority of liver OPN producing cells are inflammatory cells (Figs 1D and 2C; S2 Fig), while the ductular reaction is the most important source of OPN in chronic schistosomiasis (Fig 2C and 2E). OPN expression in both human and murine acute schistosomiasis is enriched in the macrophage area of the necrotic-exudative granulomas. Double immunohistochemistry for OPN and CD68 (a macrophage marker) confirmed that the macrophages in acute schistosomiasis express this pro-inflammatory cytokine (Fig 3A). Since the macrophages are in contact with egg antigens, we investigated if soluble egg antigens could stimulate OPN production in vitro. Primary human Kupffer cells incubated with SEA for 3 hours upregulated OPN mRNA (p = 0. 0082) (Fig 3B), indicating that infection per se can directly increase macrophage expression of this proinflammatory and profibrogenic molecule. We demonstrated for the first time that circulating osteopontin levels are increased in human acute schistosomiasis mansoni. Our results also suggest that serum OPN measurement could be a good biomarker to diagnose symptomatic acute schistosomiasis. The highest levels of OPN were observed in patients during the peak of clinical symptoms (7–11 weeks post infection). Once the granulomas were modulated (>12 weeks post infection) the OPN levels decrease significantly. Circulating and hepatic OPN levels were also elevated in the acute phase of experimental murine schistosomiasis mansoni. Chen et al. (2011) demonstrated that liver OPN levels peaked at the acute phase of S. japonicum infection. As previously mentioned, the murine model has some limitations in regard to acute schistosomiasis [14]. However the model may be helpful to identify the factors related to the onset of the generalized reactive changes during the early course of a primary schistosomal infection [14]. Importantly, our new data in humans demonstrate that the mouse model mirrored the human disease with regards to the pattern of OPN expression, reinforcing that this model could be useful to understand the mechanisms related to the acute phase of schistosomiasis in humans. Macrophages are the major OPN producing cell in acute schistosomiasis and SEA induces OPN expression in primary human Kupffer cells. Pereira et al. (2015) also observed that macrophages are one of the major sources of OPN in the early phases of infection in mice and in patients with hepatointestinal schistosomiasis, while bile ducts are the main producers of OPN in patients with hepatosplenic disease. We confirm that osteopontin is mostly expressed by the ductular reaction in mice in the late chronic phase of infection. Pereira et al. (2015) also observed that SEA stimulates primary mouse Kupffer cells, stellate cells and cholangiocytes to produce OPN, demonstrating that egg antigens directly induce the expression of this pro-inflammatory and pro-fibrogenic molecule by multiple types of cells that localize in schistosoma-infected livers. Osteopontin has been previously associated with acute hepatic injury [16,17,22,23]. Patients with acute liver failure of different etiologies such as acetominophen toxicity, ischemia (shock), idiosyncratic drug-induced liver injury, autoimmune hepatitis and viral hepatitis A and B, have increased OPN plasma levels [22,23]. Recent findings indicate that OPN plays a central role in liver diseases associated with necrosis [16,17,23]. Liver injury triggers OPN production in Kupffer cells and NKT cells that attract neutrophils, lymphocytes and macrophages to affected areas [16,17,19,24]. The recruited cells become activated and produce OPN and Th1 cytokines, exacerbating liver necrosis [16,17,19,24]. In acute liver failure patients, OPN was particularly associated with hyperactute injury [23]. The role of OPN has been described in granulomatous reactions, especially Th1-mediated, [16,19]. OPN is essential for Th1 polarization [25] and OPN from dendritic cells mediates granuloma formation against bacterial antigens [26]. OPN expression in sarcoidosis, tuberculosis and other Th1-mediated granulomas is more associated with macrophages than extracellular matrix [27]. Using the B-glucan model, Morimoto et al. (2004) demonstrated that OPN-/- mice have a reduction in granuloma size and number and a 2-fold decrease in macrophage accumulation [28]. Overexpression of OPN increased granuloma formation and delayed its resolution, promoting an exacerbated fibrotic response [28]. Similar findings were observed by O’Regan and coworkers (2008) in S. mansoni egg-induced lung granulomas, a typical Th2-mediated granuloma [20]. Our results confirm the pivotal role of OPN in the Th1 and Th2 mediated granulomas and demonstrate that pathogen antigens directly induce OPN production by macrophages. Acute schistosomiasis is a systemic hypersensitivity reaction against S. mansoni and it is characterized by miliary distribution of hyperergic necrotic-exudative granulomas [2]. The live miracidia inside the egg secrete a series of antigens and lytic substances that can trigger OPN production, recruiting inflammatory cells and inducing the granulomatous reaction to prevent further liver damage (Th1 over Th2 response) [2,3, 10,18]. As disease progress (Th2 over Th1 response), the granulomas are modulated (decrease in IFN-gamma and increase in IL10), the antigens and lytic substances are sequestered, necrosis is no longer observed and OPN is down regulated [2,3, 10,12]. Patients that will develop hepatosplenic schistosomiasis continue to produce OPN, especially by the ductular reaction, promoting fibrosis and portal hypertension [18]. The plasma levels in acute schistosomiasis were even higher than observed in hepatosplenic patients. Although OPN was demonstrated to be stable in both serum and plasma, OPN levels in the serum are 3. 8–4. 8 times lower than in plasma [29]. The authors speculate that this phenomenon may reflect OPN sequestration by the clot or its cleavage by thrombin, leading to loss of immunoreactivity [29]. In our cohort of acute patients only a small number of individuals had both plasma and serum samples collected and we also observed a 4–4. 5 times reduction of OPN levels in serum compared to plasma (S1 Table). Ideally, future studies should use plasma samples in order to measure the total amount of circulating osteopontin. In conclusion, S. mansoni egg antigens induce the production of OPN by macrophages in the necrotic-exudative granulomas characteristic of acute schistosomiasis mansoni. Circulating OPN levels are upregulated in human and murine acute schistosomiasis and could be a non-invasive biomarker of this form of disease.

Title: Osteopontin Is Upregulated in Human and Murine Acute Schistosomiasis Mansoni Summary: Schistosomiasis is a major health problem that affects over 200 million people. Symptomatic acute schistosomiasis is a systemic reaction to the worms and eggs in individuals from non-endemic areas after a primary infection. Tourists, military personnel and people who practice water sports are at risk. Although most cases resolve 90 days post infection, severe cases with massive distribution of eggs can be fatal. It is frequently misdiagnosed, under diagnosed or has delayed diagnosis because the signs and symptoms are nonspecific and eggs are usually present in stool only 6 weeks post-infection. The mechanisms underlying the pathogenesis of acute schistosomiasis are not fully elucidated and currently there is a lack of noninvasive biomarkers to diagnose this form of disease. We report that serum osteopontin levels are increased in patients with acute schistosomiasis and parallel the clinical symptoms, returning to baseline level once the granulomas were modulated and the symptoms resolve. Soluble egg antigens provoke macrophages to produce osteopontin, recruiting more macrophages to the site of injury and inducing the granulomatous reaction. This observation suggests that osteopontin plays an important role in acute schistosomiasis mansoni and could be a novel non-invasive biomarker for this form of the disease.

lay_plos

en

Write a title and summarize: The larva of cestodes belonging to the Echinococcus granulosus sensu lato (s. l.) complex causes cystic echinococcosis (CE). It is a globally distributed zoonosis with significant economic and public health impact. The most immunogenic and specific Echinococcus-genus antigen for human CE diagnosis is antigen B (AgB), an abundant lipoprotein of the hydatid cyst fluid (HF). The AgB protein moiety (apolipoprotein) is encoded by five genes (AgB1-AgB5), which generate mature 8 kDa proteins (AgB8/1-AgB8/5). These genes seem to be differentially expressed among Echinococcus species. Since AgB immunogenicity lies on its protein moiety, differences in AgB expression within E. granulosus s. l. complex might have diagnostic and epidemiological relevance for discriminating the contribution of distinct species to human CE. Interestingly, AgB2 was proposed as a pseudogene in E. canadensis, which is the second most common cause of human CE, but proteomic studies for verifying it have not been performed yet. Herein, we analysed the protein and lipid composition of AgB obtained from fertile HF of swine origin (E. canadensis G7 genotype). AgB apolipoproteins were identified and quantified using mass spectrometry tools. Results showed that AgB8/1 was the major protein component, representing 71% of total AgB apolipoproteins, followed by AgB8/4 (15. 5%), AgB8/3 (13. 2%) and AgB8/5 (0. 3%). AgB8/2 was not detected. As a methodological control, a parallel analysis detected all AgB apolipoproteins in bovine fertile HF (G1/3/5 genotypes). Overall, E. canadensis AgB comprised mostly AgB8/1 together with a heterogeneous mixture of lipids, and AgB8/2 was not detected despite using high sensitivity proteomic techniques. This endorses genomic data supporting that AgB2 behaves as a pseudogene in G7 genotype. Since recombinant AgB8/2 has been found to be diagnostically valuable for human CE, our findings indicate that its use as antigen in immunoassays could contribute to false negative results in areas where E. canadensis circulates. Furthermore, the presence of anti-AgB8/2 antibodies in serum may represent a useful parameter to rule out E. canadensis infection when human CE is diagnosed. The larval stage (metacestode) of Echinococcus granulosus sensu lato (s. l.) causes cystic echinococcosis (CE, traditionally referred to as hydatid disease), one of the most important and widespread parasitic zoonoses. It is a fluid-filled cyst that establishes and grows in the host viscera (mainly liver and lung) of several ungulate livestock (among others sheep, cattle, horse, goat, and pig) and wild animals [1]. Recently, phylogenetic studies have led to split E. granulosus s. l. into five species, showing preference for infecting different hosts: E. granulosus sensu stricto (including G1-G3 genotypes), E. equinus (G4), E. ortleppi (G5), E. canadensis (G6–G10) and E. felidis [2,3]. These species seem to diverge in their transmission dynamics, morphology, rate of development, antigenicity, sensitivity to drugs and, particularly, in their infectivity and pathogenicity in humans, which might therefore influence the design of therapeutic and prophylactic programmes for CE control. This emphasises the need of studies focused on the molecular characterisation and the geographical distribution of E. granulosus s. l. species/genotypes. E. granulosus sensu stricto (s. s.) uses mostly sheep as intermediate hosts, but is also capable of infecting other livestock such as cattle as well as humans. Epidemiological studies for examining E. granulosus s. l. species associated with human CE have determined that E. granulosus s. s. has an extensive geographical distribution and causes between 73% and 88% of human CE worldwide (reviewed by [4,5]). On the other hand, E. canadensis G6 and G7 genotypes, which use mainly camels, goats and pigs as intermediate hosts, are also geographically widely distributed and ranked as the second cause of human CE in the world, being responsible for between 11% and 21% of human CE cases according to more recent studies [4–6]. However, these values may be underestimated since E. canadensis seems to exhibit a lower and/or slower growth than E. granulosus s. s. in humans, leading to more benign or asymptomatic infections [3,4]. Moreover, in countries such as Austria, Poland, Egypt and Sudan, E. canadensis is the predominant cause of human CE [3]. Regarding E. canadensis genotypes, G6 has been preferably associated with human CE but, a recent systematic revision of the species and genotypes of E. granulosus s. l. responsible for human infections suggests a scenario with a slightly lower prevalence rate for G7 comparing to G6 (9. 6% vs 12. 2%, respectively) [5]. Interestingly, the geographical distribution of these genotypes differ; G6 genotype is mainly present in human CE cases from America, Asia and Africa whereas the G7 genotype seems to affect mostly some countries in Central Europe. It is worth to mention that there is little or no genotype information on human CE cases reported in many geographical regions/countries, which might influence the epidemiological data cited above. Despite some progress achieved by prevention campaigns, CE continues being a major public health problem in several countries while represents an emerging or re-emerging disease in others (reviewed by [7,8] and [9–13]). Regarding CE diagnosis, antigen B (AgB), an abundant parasite component present in the HF of the E. granulosus s. l. metacestode, is the most immunogenic and specific Echinococcus-genus antigen. It is a 230 kDa lipoprotein that carries a huge amount of both neutral and polar lipids (around 50% in mass) including fatty acids (FA) and sterols, which Echinococcus is not capable of synthesising (reviewed by [14]). This has led to emphasise its hypothetical role in parasite lipid metabolism, taking up host lipids as building blocks for parasite metabolic demands. Moreover, this hypothesis is supported by the fact that AgB belongs to a cestode-specific family of proteins exhibiting ability to bind hydrophobic ligands (HLBP for hydrophobic ligand binding protein) [15,16]. This family has emerged by independent gene expansion events, giving rise to species and gene-specific monophyletic clades. Interestingly, HLBP members are all immunodominant antigens. AgB antigenicity has been associated with its protein moiety (apolipoprotein components) [17–19] that is encoded by a multigene and polymorphic family with five AgB gene products named AgB1 to AgB5 (revised by [20]). The recent assembly of Echinococcus granulosus G1 genotype and E. multilocularis genomes confirmed that this scenario is highly conserved among Echinococcus species [21]. The mature protein products of these genes are small (around 8 kDa in mass), α helix-rich secreted polypeptides, with ability to self-assembly generating high-molecular-mass oligomers [22,23]; they are thus named AgB8/1 to AgB8/5 subunits. The native antigen, the recombinant AgB8/1 and AgB8/2 subunits, as well as various synthetic peptides derived from them, have shown to be valuable for CE diagnosis [24–26]; all of them have shown similar diagnostic performance in comparison with crude HF preparations, but in some clinical studies recombinant AgB8 subunits (rAgB8) seem to yield better specificity with little or no loss in sensitivity [27–30]. AgB gene expression in Echinococcus s. l. species has been examined suggesting differences between them; this might be relevant for epidemiological investigations intended to discriminate the contribution of distinct E. granulosus s. l. species to human CE. In the larva of E. granulosus s. s. all AgB genes were found to be expressed at mRNA level [31], even though only AgB8/1 to AgB8/4 protein products have been certainly detected in HF [23]. On the other hand, no evidence of AgB5 expression or of the generation of AgB8/2 and AgB8/5 was achieved in E. canadensis (G6 and G7 genotypes) and E. ortleppi (G5 genotype) metacestode [32,33]. In particular, AgB2 was proposed to be a pseudogene in E. canadensis. In fact, a low-scale sequencing analysis of E. canadensis genomic DNA, revealed that AgB2-related sequences (named EgB2G6v15 to EgB2G6v17 and EgB2G7v15, EgB2G7v18 and EgB2G7v19) contained a substitution at the splicing site (GT-TG instead of GT-AG) that probably interferes with the splicing, leading to the formation of a premature stop codon [32,33]. Taking advantage of the recently available genome of E. canadensis G7 genotype (published at http: //parasite. wormbase. org as echinococcus_canadensis. PRJEB8992. WBPS5. protein), we confirmed the existence of this substitution in ECANG7_10984, which corresponds to the first hit by Blastn analysis using the E. granulosus s. s. AgB2 sequence Q27275 as a query at the http: //parasite. wormbase. org webpage. However, the generation of a functional AgB2 product may occur by a non-canonical transcriptional mechanism using the TG dinucleotide as splice acceptor site [34]. Studies at transcriptional level failed to identify mRNA coding for a functional AgB2 product in E. canadensis G7; detected AgB2 mRNA transcripts were compatible with the use of an upstream AG dinucleotide in the second exon as splice acceptor site that would yield a protein considerably shorter than AgB8/2 due to a premature stop codon [33]. Nevertheless, these studies were carried out using protoscoleces derived from a single cyst of G7 origin (Muzulin et al, 2008), and the germinal layer constitutes a metacestode structure relevant in terms of AgB expression. On the other hand, a deep-sequencing analysis of the transcriptome of E. canadensis G7 metacestode has not been performed yet. Taken together, the conversion of AgB2 into a mature and functional product in the larva of E. canadensis remains uncertain and has not been explored using proteomic tools yet. It is important to remark that predictions based on draft genomes and transcriptional studies are not the ultimate proof of the absence or presence of a protein. Post-transcriptional control of gene expression could play an important role; a gene with low or undetectable expression at the transcriptional level could be efficiently translated allowing the detection of the encoded protein. Various proteomic studies have analysed the parasite and host components present in the HF of E. granulosus s. l. [23,35,36], nevertheless, none of them provide data about E. canadensis AgB. In this work, we have employed high sensitivity proteomic tools to determine the apolipoprotein composition of AgB present in the HF of E. canadensis G7 genotype. For this proteomic study, we used swine HF as a source of AgB because pigs constitute the main intermediate hosts for E. canadensis G7 genotype, and HF collects products secreted/excreted by the germinal layer as well as protoscoleces, representing the parasite material where AgB accumulates. Complementary and high-sensitivity approaches including two-dimensional gel electrophoresis (2-DGE) and liquid chromatography (LC) coupled to mass spectrometry (MS) were used as proteomic tools. For a complete biochemical characterisation of E. canadensis AgB, the lipids carried by the lipoprotein were also examined by high performance thin layer chromatography (HPTLC). Results highlight the concept that AgB is a complex lipoprotein in E. granulosus s. l. species, including E. canadensis, being AgB8/1 the predominant apolipoprotein. Furthermore, in contrast with E. granulosus s. s. [23], AgB8/2 was not detected in E. canadensis G7 genotype, supporting the concept that AgB2 is a pseudogene in this species. Since AgB is the most relevant antigen for human CE immunodiagnosis, and the use of rAgB8 subunits offers several advantages for standardising immunoassays (reviewed by [26]), the possible implications of our findings on diagnostic and epidemiological studies on human CE are discussed. Fertile hydatid cysts (containing protoscolex, n = 24) were collected from livers of naturally infected pigs during the routine work of local abattoirs in Buenos Aires (Argentina). HF was obtained by aspiration of the content of cysts, and preserved by addition of 5 mM EDTA and 20 μM 3,5-di-tert-butyl-4-hydroxytoluene (BHT) at -20°C until use. Protoscolex were used to analyse parasite genotype on individual cysts. Cyst genotyping was performed by amplification and sequencing of a fragment of the mitochondrial cytochrome c oxidase subunit 1 (COX1) [37]. The sequencing reactions were performed at Macrogen (Korea). All HF samples of swine origin were confirmed to belong to E. canadensis G7 genotype. For controlling the sensitivity of our proteomics tools, we prepared a pool of bovine HF samples (similarly obtained from local abattoirs in Montevideo, Uruguay). This bovine pool was mainly representative of E. granulosus s. s. as it contained material from 20 and 3 cysts belonging to E. granulosus s. s. (18 of G1 and 2 of G3 genotypes) and of E. ortleppi (G5 genotype), respectively. An AgB-enriched fraction was prepared from pooled HF by removing the bulk of host albumin and immunoglobulins by anion exchange chromatography. HF was centrifuged at 10000 x g for 20 min at 4°C and the resulting supernatant filtered through 0. 45 μm filter membranes (Millipore). The clarified HF (700 mL) was then fractioned by anion exchange chromatography on a Q-Sepharose column (2. 5 cm x 10 cm, Pharmacia Biotech, Uppsala, Sweden) previously equilibrated in 20 mM phosphate buffer, pH 7. 4 containing 200 mM NaCl, 5 mM EDTA and 20 μM BHT. After washing in equilibration buffer, the retained material was eluted by changing ionic strength to 500 mM NaCl in a single step. The eluted fraction, Q-Sepharose retained fraction (QSf), was concentrated 10-times, equilibrated in 20 mM phosphate buffer, pH 7. 4 containing 150 mM NaCl, 5 mM EDTA and 20 μM BHT (PBSEDTA-BHT), and used to characterise AgB apolipoprotein composition by mass spectrometry as described below. A second purification step was performed based on ultracentrifugation of QSf in a KBr density gradient. Briefly, 2. 45 g of KBr were dissolved in 5 ml of QSf in an ultracentrifuge tube and slowly covered with a solution containing 0. 15 M NaCl and 0. 42 M KBr. After ultracentrifugation (4 h at 332. 000 x g) two bands were carefully recovered named low (Ldf, yellowish-brown band) and high (Hdf) density fractions. All fractions were equilibrated in PBSEDTA-BHT, and maintained at 4°C under a N2 atmosphere until use. 2-DGE and MS analysis was performed as described previously [38] but using 150 μg (protein) of the AgB-enriched fraction (QSf) for the electrofocusing step in order to detect poorly represented subunits. Briefly, the first dimension was performed with commercially available IPG-strips (7 cm, linear 3–10, GE Healthcare). QSf was prepared and concentrated by using the 2-D Clean-Up kit (GE Healthcare) and dissolved in rehydration solution (7 M urea, 2 M thiourea, 2% CHAPS, 0. 5% IPG buffer 3–10 (GE Healthcare), 0. 002% bromophenol blue, 17 mM DTT). Samples in rehydration solution were loaded onto IPG-strips by passive rehydration during 16 h at room temperature. The second-dimensional separation (SDS-PAGE) was performed in 15% polyacrylamide gels using a SE 260 mini-vertical gel electrophoresis unit (GE Healthcare). The molecular size marker used was Low Molecular Weight Calibration Kit for SDS Electrophoresis (Amersham GE Healthcare). The gels were colloidal coomassie stained and images were digitalised using a UMAX Power-Look 1120 scanner and LabScan 5. 0 software (GE Healthcare). Selected spots were submitted to in gel trypsin digestion (sequencing-grade, Promega) at 37°C overnight. Peptides were extracted from gels using 60% acetonitrile in 0. 1% TFA, concentrated by vacuum drying, and then desalted using C18 reverse phase micro-columns (OMIX Pipette tips, Varian). Peptide elution from micro-column was performed directly into the mass spectrometer sample plate with 2 μl of matrix solution (α-cyano-4-hydroxycinnamic acid in 60% aqueous acetonitrile containing 0. 1% TFA). Mass spectra of digestion mixtures were acquired using a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF/TOF, 4800 Analyzer, ABi Sciex) in positive reflector mode and were externally calibrated using a mixture of peptide standards (Mix 1, ABi Sciex). Collision induced dissociation (CID) MS/MS spectra of selected peptides ions were also acquired. Proteins were identified with measured m/z values in MS and MS/MS acquisition modes and using the MASCOT search engine (Matrix Science, http: //www. matrixscience. com) in the Sequence Query search mode. AgB8 subunits were identified by searching in both, the NCBInr and an in-house Echinococcus databases using the following search parameters: unrestricted taxonomy, monoisotopic mass tolerance, 0. 05 Da; fragment mass tolerance, 0. 2 Da; carbamidomethyl cysteine and methionine oxidation as variable modifications and up to one missed tryptic cleavage allowed. Significant protein scores (p < 0. 05) were used as criteria for positive protein identification. In addition, at least two unique peptides with ion significant score (p < 0. 05) were required for AgB8 subunit identification. The in-house Echinococcus database was built comprising all sequences of E. canadensis (G7 genotype, published in http: //parasite. wormbase. org as echinococcus_canadensis. PRJEB8992. WBPS5. protein) and of E. granulosus s. s. (G1 genotype, published in www. genedb. org as EGU_proteins_29042013_products. fa) plus a total of 102 full length sequences, including polymorphic variations at the level of the AgB mature products as well as the orthologous products in other Echinococcus species (available on NCBInr, March 2015). Furthermore, to study E. canadensis AgB8/2 presence, we took into account the previous characterisation of this gene (at DNA and mRNA level, [32]) and added to the database those protein sequences that would be generated by non-canonical splicing of E. canadensis AgB2-related sequences EgB2G6v15 to EgB2G6v17, EgB2G7v15, EgB2G7v18 and EgB2G7v19, as well as of AgB ECANG7_10984 gene (in all putative open reading frames, S1 Appendix). Samples (QSf) were analysed by LC tandem-mass spectrometry (LC-MS/MS) using five analytical replicates. Proteins were reduced, carbamidomethylated, and digested in solution with sequencing-grade trypsin (Sequencing-grade Promega; 1: 50 enzyme to total protein ratio) in 70 mM ammonium bicarbonate pH 8. 0 buffer containing 2 M guanidine hydrochloride for 12 h at 37°C. Peptides were further concentrated, desalted using C18 reverse phase micro-columns (OMIX Pipette tips, Varian) and eluted with 60% aqueous acetonitrile containing 0. 1% TFA. Peptide mixtures were dried and resuspended in 5% aqueous acetonitrile containing 0. 1% formic acid. Five micrograms of each sample were analysed in an EASY-nLC II nanoflow liquid chromatography (Thermo Fisher Scientific, USA) coupled to a LTQ-Orbitrap Velos mass spectrometer (Thermo Fisher Scientific). Peptide mixture was injected into a trap column (I. D. 100 μm x O. D. 360 μm x 50 mm) packed with Jupiter C18 10 μm beads (Phenomenex Inc., USA) for desalting with 100% solvent A (0. 1% formic acid). Peptides were then fractionated on an analytical column (I. D. 75 μm x O. D. 360 μm x 100 mm) packed in-house with Aqua C-18 5 μm beads (Phenomenex Inc.) at a flow rate of 200 nL/min using a 60 min linear gradient from 5 to 35% of solvent B (0. 1% formic acid in acetonitrile). Afterwards, a gradient from 35 to 85% of B in 5 min was applied for ensuring a complete elution. Nano-electrospray voltage was set to 2. 3 kV, the source temperature to 250°C and mass spectrometer was operated in a data-dependent acquisition mode, where the top ten precursor ions in each cycle were selected for fragmentation event by CID. Ion trap injection time was set to 100 ms and FT-MS injection time was set to 1000 ms with a resolution of 60,000 across m/z 300–1800. For IT scans, fragmentation was carried out on ions above a threshold of 200 counts, and dynamic exclusion was enable with an exclusion list size of 500 for 90 seconds, repeat duration of 30 seconds and a repeat count of 1. Raw mass data files (. raw) were analysed in Maxquant (v. 1. 5. 5. 1) and its built-in Andromeda search engine. Parasite and host proteins were identified using MaxQuant software by searching MS and MS/MS data against a merged database comprising the Echinococcus database (built as described above) and the Bos taurus/Sus scrofa database (downloaded from UniProt, April/2016). Trypsin was set for enzyme specificity with a maximum of two missed cleavages, mass tolerance for precursor ions was set to 10ppm, and fragment ion mass tolerance was set to 0. 5 Da. MS/MS spectra searches incorporated fixed modifications of carbamidomethylation of cysteine, oxidation of methionine and protein N-terminal acetylation were set for variable modifications. Maximum false peptide and protein discovery rate was set to 0. 01. Proteins matching to the reverse database were eliminated. Statistical analysis for protein identification was performed using Perseus (v. 1. 4. 0. 11) based on unique peptides MS intensities, the presence of a minimum of two unique peptides and PEP (posterior error probability) < 0. 01. To evaluate the abundance of each AgB protein species (AgB8 subunit) the intensity-based absolute quantification (iBAQ) was used as it has been reported as a useful label-free quantification method provided by MaxQuant. In the iBAQ algorithm the sum of all identified peptide intensities (maximum detector peak intensities of the peptide elution profile, including all peaks in the isotope cluster) is divided by the number of theoretically observable tryptic peptides, and expressed as log2 values [39]; this operation transforms a measure that is expected to be proportional to mass (intensity) into one that is proportional to molar amount (iBAQ). To determine the relative abundance of each AgB8 subunit in AgB (riBAQAgB), we divided the iBAQ value corresponding to each AgB8 subunit by the sum of the iBAQ values obtained for all AgB8 subunits, and expressed this ratio as a percentage. Replicate results were merged with Perseus and values for iBAQ, riBAQAgB, score and the percentage of the protein sequences covered by identified peptides (% CO) are expressed as the mean of all runs (n = 5). The total number of the identified peptide spectra matched for a protein (PSM) was also estimated as the sum of all runs. Only proteins present in at least 3 of the 5 analytical replicates were considered as positively identified. AgB total lipids were analysed using Ldf (between 0. 25 and 0. 5 mg of protein) following the methodology that we have already described [38]. Qualitative analysis of lipid classes was performed by HPTLC using double development for neutral and polar lipids as described previously [38], but lipid bands were visualised under iodine vapour. Identification of lipid classes was performed by comparison with primary and secondary standards run on the same HPTLC plate. The apolipoprotein composition of AgB present in fertile HF of E. canadensis G7 genotype was analysed using MS based methodologies. For this purpose, we prepared a biological representative pool of E. canadensis HF from 24 individual swine cysts, each one of G7 origin according to COX1 genotyping. However, to achieve an adequate sample for AgB apolipoprotein characterisation, we firstly carried out an enrichment step since AgB is poorly represented in HF compared to host albumin and immunoglobulins. Taking advantage that AgB can be selectively separated from these host proteins employing a Mono-Q [40] or Q-Sepharose beads [41], we prepared an AgB enriched-fraction by a single step anion exchange chromatography of HF on Q-Sepharose; this step concentrates AgB favouring the detection of lower represented apolipoproteins. S1 Fig shows the SDS-PAGE analysis of fractions obtained by this chromatography. As expected, AgB was retained by Q-Sepharose beads and eluted with 500 mM NaCl pH = 7. 4 (fraction QSf) since QSf, but not the flow through fraction (FTf), showed 8,16 and 24 kDa bands in agreement with the typical AgB ladder-like pattern (S1 Fig, small head arrows) [42]. In contrast, the majority of the most abundant host proteins present in HF (albumin and immunoglobulins) did not bind to Q-Sepharose, being recovered in FTf (S1 Fig). Additional steps based on ultracentrifugation on a KBr density gradient achieved to purify AgB (see below). Nevertheless, since these purification steps led to protein losses, we rather to use QSf for characterising AgB protein species; we cannot ruled out that AgB includes particles of different densities and/or less abundant AgB apolipoproteins would be not represented in the purified AgB preparation (Ldf, see below). The presence of AgB8 subunits in swine QSf (sQSf) was examined by 2-DGE followed by MALDI-TOF/TOF MS. As shown in Fig 1A, AgB was detected in several spots corresponding to the monomer, dimer and trimer (indicated with bold circles and numbers in the Fig). Interestingly, the monomeric as well as oligomeric AgB forms comprised several components spread over a wide range of pH (between 9. 4 and 4. 5). AgB protein species identified in those spots included AgB8/1, AgB8/3 and AgB8/4 subunits (Fig 1A and S1 Table). AgB8/1 was the predominant subunit detected in all spots belonging to AgB (n = 25); the presence of both Q86BY8 and Q3YFQ5 isoforms is plausible accordingly to the set of unique peptides identified by MALDI-TOF/TOF (Table 1). Q86BY8 and/or Q3YFQ5 are basic proteins (pI = 9. 11) that would agree with their identification in spots focused at around pH 9. 4 (named #1 and #2, Fig 1A), but not in more acidic ones. Similarly, two AgB8/4 protein species with a theoretical pI of 6. 15 (named Q6J0W7 and Q6Q0G2, Table 1) were detected in 9 spots focused in a wide range of pH (Fig 1A and S1 Table). Thus, these results suggest the presence of post-translational modifications in both AgB8/1 and AgB8/4. Neither signals corresponding to phosphorylated peptides nor the formation of carbonyl groups in AgB8 subunits (because of oxidative reactions with oxides of nitrogen or metal catalysed oxidation) were detected by MS and Western Blot, respectively (S2 Appendix). Thus, further studies are needed to elucidate which molecular modifications explain the AgB pattern obtained by 2-DGE. On the other hand, AgB8/3 was identified in spots #1 and #2 based on various unique peptides (Table 1 and S1 Table); while these peptides cannot distinguish between the AgB8/3 isoforms named Q6VXZ8 and Q6VXZ9, the presence of Q6VXZ8 seems to be more likely according to its pI. Finally, AgB8/2 and AgB8/5 were not detected in sQSf. The presence of AgB8/2, but not AgB8/5, has been previously reported in bovine HF collected from E. granulosus s. s. cysts [23]. We performed thus a similar study by 2-DGE plus MALDI-TOF/TOF using a pool of bovine HF (mostly belonging to E. granulosus s. s. according to COX1 genotyping) to evaluate whether our proteomic tool reached enough sensitivity for AgB8/2 analysis. Results showed the presence of AgB8/2, but not of AgB8/5 in bovine QSf (bQSf, Table 1, S3 Appendix) in accordance with the earlier report. Overall, detection of AgB8/2 in bQSf but not in sQSf suggests that AgB8/2 is not present in G7-HF. In addition to AgB protein species, analysis of sQSf and bQSf by 2-DGE plus MALDI-TOF/TOF showed the presence of Echinococcus Ag5 (22 and 38 KDa subunits) as well as of some host components (remaining albumin and immunoglobulin light chains, as well as apolipoprotein A-I (Apo A-I); see S1 Table and S3 Appendix). Confirmation of the observations described above was achieved using LC-MS/MS since this methodology enables a high sensitivity quantitation of proteins in complex biological samples. Results are summarised in Table 2, in which the iBAQ parameter (expressed as log2 values) is proportional to the protein molar amounts while the riBAQAgB refers to the relative abundance of each protein species in AgB. Analysis of sQSf showed that AgB8/1 was the major protein component, representing 71% of total AgB apolipoproteins followed by AgB8/4 (15. 5%), AgB8/3 (13. 2%) and AgB8/5 (0. 3%). AgB8/2 was not detected in sQSf although our database included all AgB8/2 sequences available for E. granulosus s. l. species (comprising those protein products that could be generated because of non-canonical splicing of all available AgB2 related sequences). As expected, E. granulosus s. s. AgB8/2 was identified in bQSf. Because the bovine HF pool contained samples from E. ortleppi G5 genotype, we looked for E. ortleppi AgB8/2 specific peptides in bQSf with no success. This may be consequence of the low proportion of E. ortleppi components in the bovine HF pool (around 15% of the total volume), or of the lack of AgB2 functionality in this species, as proposed previously [32]. On the other hand, we detected in sQSf two unique peptides that make reliable the identification of AgB8/5 in E. canadensis metacestode (Table 2), contrasting with previous findings at the RNA level [33]. This contrast can be explained by the fact that AgB5 would be poorly expressed in the metacestode and/or that previous mRNA expression studies were performed using primers, which were not specifically designed for E. canadensis AgB5. On the other hand, our results endorse previous data at mRNA [31] and protein levels [35] for AgB5 expression in E. granulosus s. l. metacestode. Finally, the detection in sQSf of AgB8/5, an AgB subunit barely expressed in the metacestode of Echinococcus species, denotes the high sensitivity reached in our proteomic study. AgB is likely involved in taking up host lipids, which are essential for Echinococcus spp., as building blocks for parasite needs [14]. For a complete biochemical characterisation, we purified E. canadensis AgB from QSf and characterised the lipid classes present in its lipid moiety. AgB purification was performed by a novel procedure based on density-gradient ultracentrifugation; this method preserves AgB native structure and yields AgB particles independently of its apolipoprotein composition. AgB was mainly recovered in the low density fraction, Ldf, but consecutive ultracentrifugation rounds were needed to achieve a good-quality AgB preparation (about 95% pure, according to SDS-PAGE, S1B Fig), although these steps goes against the final AgB yield. Several lipid classes including highly polar (phosphatidylcholine and, to a lesser extent, phosphatidylethanolamine) and neutral lipids (sterols, free FA, triacylglycerols and sterol esters) were detected in E. canadensis AgB (Fig 1B), just as we have already described for AgB immunopurified from bovine pooled HF ([38] and S3 Appendix). In sum, the observed differences in the protein composition of AgB preparations from distinct E. granulosus s. l. species (Table 2), did not affect the lipid class composition of the lipoprotein. However, qualitative and or quantitative differences in lipid components within each class cannot be excluded and require further studies. The proteomic analysis of sQSf and bQSf by LC-MS/MS allowed identifying several parasite and host proteins in HF (S4 Appendix). Regarding parasite proteins, we identified several proteins with putative diverse functions, but taking into account the iBAQ values in both samples, AgB subunits were found to be the most abundant components, followed by Ag5. In particular, we significantly identified in sQSf and/or bQSf parasite proteins with a potential role in lipid metabolism. One of the most interesting was an E. granulosus s. s. HLBP detected in bQSf (EgHLBP, Uniprot protein accession A0A068WMS7_EGHR). The gene that encodes EgHLBP, referred to as EgrG_000549200 (http: //www. genedb. org/), mapped outside AgB cluster [43]. This novel EgHLBP has a higher similarity to an uncharacterised protein of E. granulosus s. s. (Uniprot protein accession W6UNU2_ECHGR, 87% identity) and to Taenia solium HLBP1 and HLBP2 (Uniprot protein accession G3FJ94_TAESO and G3FJ95_TAESO, with 68% and 71% of identity, respectively) than to AgB (45% identity); the alignments of EgHLBP with these proteins are shown in S5 Appendix. Interestingly, we detected EgHLBP in HF, while both TsHLBP1 and TsHLBP2 were not detected by Western Blot in the metacestode and showed a high expression in the T. solium adult [44]. In addition, we identified the Lipid transport protein N-terminal in both sQSf and bQSf. This protein exhibits only significant similarity with its orthologous in E. multilocularis (96% identity) and with an E. granulosus s. s. apolipophorin (Uniprot accession W6UHB7_ECHGR, 98% identity), neither of which have been characterised yet. All of them are high MW macromolecules, containing various conserved regions found in several lipid transport proteins, including vitellogenin, microsomal triglyceride transfer protein and apolipoprotein B-100 (Smart accession number SM00638; PROSITE PS51211). Finally, we identified various host apolipoproteins, including Apo A-I, in sQSf and bQSf. Apo A-I has previously been detected in E. granulosus s. l. HF [35]. Interestingly, Apo A-I and an Apo A-I binding protein (EmABP) were found to be present in HF of E. multilocularis [45]. In our study, EmABP orthologues in E. granulosus s. l. species were not found in QSf or bQSf, but their presence in HF requires further investigation. This work contributes to widen the information available on E. granulosus s. l. AgB subfamilies, particularly at the level of the presence and abundance of their protein products in the metacestode of E. canadensis G7 genotype. Using high sensitivity and quantitative proteomic analysis of a representative number of hydatid cysts, we showed that AgB8/1 is the major AgB apolipoprotein in the HF of E. canadensis G7 genotype. This strengthens the concept of AgB8/1 predominance in the HF of various E. granulosus s. l. species [23,35]. Since AgB likely contributes to the mechanisms used by the metacestode to transport lipids, particularly those that the parasite is unable to synthesise, this result would indicate that AgB8/1 is the main AgB apolipoprotein involved in this transport, and, in consequence, the presence of AgB8/1 receptors in parasite and host cells is worth to be further studied. To this respect, AgB8/1 was found to bind selectively to monocyte and macrophages, but the molecular partners involved have not been identified yet [41]. Furthermore, we identified an additional Echinococcus HLBP (EgrG_000549200) and host apolipoproteins (particularly Apo A-I) in QSf, which suggests that several lipid carriers are involved in parasite mechanisms aimed at providing essential lipids to metacestode tissues. However, taken into account their abundance in HF (iBAQ values), their contribution to lipid transport within metacestode tissues seems to be lower than that of AgB. On the other hand, our results support a differential expression of AgB2 among E. granulosus s. l. species; AgB8/2 was not detected in the HF of E. canadensis G7 genotype contrasting with their detection in E. granulosus G1 genotype ([23] and this work). Despite this difference, we did not found significant differences in the lipid moiety of AgB purified from sQSf and bQSf, at least in terms of lipid classes. This may be a result of the fact that AgB8/2 showed a low relative abundance in comparison with AgB8/1 in E. granulosus s. s., and that no differences have been observed between the lipid binding properties of AgB8/1 and AgB8/2 using in vitro assays [46]. Regarding to the lack of AgB8/2 in E. canadensis, it would be explained at the molecular level by the occurrence of an A/T transversion at the splicing site that likely interferes with canonical splicing mechanisms and the synthesis of a functional protein product. Bearing in mind that AgB is diagnostically valuable for human CE, differences in AgB apolipoprotein composition between E. granulosus s. s. and E. canadensis are informative for diagnosis and epidemiological studies on this zoonosis. In particular, rAgB8/1 and rAgB8/2 subunits, as well as peptides derived from them, have yielded reasonable diagnostic performance in immunoassays using panels of sera from patients with CE and other helminth infections (reviewed by [26]). In most of these studies rAgB subunits were assessed as single antigens, although combination between them (AgB subunit cocktail) or with other HF antigens would help to achieve more sensitive and specific tests [27,47–51]. In any case, our results indicate that the use of AgB8/2 as antigen in immunoassays might contribute to false-negative results in patients infected by E. canadensis G7 genotype. Probably the same holds for infections with E. canadensis G6 genotype, although the lack of AgB8/2 at the protein level requires confirmation. As we have mentioned above E. canadensis accounts for between 11% and 21% of CE human cases worldwide reaching a higher prevalence in some countries, therefore, our results denote the importance of adapting the diagnostic tools to the epidemiological situation of each geographical region. Taking into account the global distribution of E. canadensis, our observations may thus be of major importance for regions where cases of human infection by E. canadensis have been reported (including countries in all continents, such as Argentina, Egypt, Iran, Kenya, Mauritania, Mongolia, Poland, South Africa and ex-Yugoeslavia, [4,5]). However, because of the scarce information about the E. granulosus species/genotypes associated with human CE in many countries/regions, our data might be of interest for any region where E. canadensis is known to circulate (for instance, Southern Brazil and the Mediterranean region [52,53]). The determination of E. granulosus s. l. genotype/species responsible for human CE cases is a subject of relevance since, as we have already mentioned, species belonging to E. granulosus complex differ in biological features (i. e. infectivity and pathogenicity in humans), influencing control program design as well as disease follow-up and treatment [54]. Genotyping is thus a critical task. However, nowadays, it can only be performed after surgery in order to obtain parasite samples, and using molecular biology tools. Since E. granulosus s. s. and E. canadensis are the most common cause of human CE, it would be worth to develop simple tools to differentiate the infections caused by them. The presence of antibodies against AgB8/2 in serum would be easy to determine through conventional immunoassays based on the use of rAgB8/2 or AgB8/2-derived peptides as antigen. Thus, including this kind of immunoassays during routine diagnosis of human CE would allow ruling out E. canadensis infection based on the presence of anti-AgB8/2 antibodies.

Title: Characterisation of Antigen B Protein Species Present in the Hydatid Cyst Fluid of Echinococcus canadensis G7 Genotype Summary: Cystic echinococcosis (CE), a worldwide-spread zoonosis, affects livestock mammals and humans with significant economic and public health impact. It is caused by the infection with the larva of cestodes belonging to Echinococcus granulosus complex, a series of parasite species with preference for different hosts. Among them, Echinococcus canadensis larva uses mainly camels, goats and pigs as hosts. Species/genotypes belonging to E. canadensis are considered the second most common cause of human CE, but its contribution may be underestimated since causes asymptomatic or more benign infections than other E. granulosus complex species. The most relevant antigen for CE diagnosis is a lipoprotein called antigen B (AgB). AgB antigenicity is linked to its protein moiety that is encoded by several genes. One of these genes, AgB2, seems to be differentially expressed within E. granulosus complex. Using high sensitivity proteomic tools we analysed the composition of AgB obtained from E. canadensis larva, detecting the protein products of all AgB genes, except AgB2. Since AgB subunits have been widely used as antigens in immunoassays for human CE diagnosis, our results indicate that using AgB2 protein product in these assays may lead to false-negative results, particularly in geographical areas where E. canadensis species/genotypes circulate.

lay_plos

en

Summarize: Probabilmente il figlio di Stella Creasy è l'individuo di sesso maschile più giovane mai entrato in Parlamento. La deputata lo ha portato con sé, a pochi giorni dal parto, rivendicando i diritti delle neo mamme e chiedendo per loro maggiori tutele e più sostegno, piuttosto che rimproveri costanti e scarso aiuto. La donna aveva già fatto un gesto simile in passato, facendo ingresso in Aula assieme alla figlia appena nata, per sensibilizzare sull'importanza di rientrare a lavoro in piena serenità, senza essere giudicate ma senza neppure essere abbandonate a se stesse. Con il supporto adeguato, conciliare carriera e famiglia è possibile. La 44enna Stella Creasy è membro del Parlamento del Regno Unito dal 2010. La laburista ha avuto una figlia nel 2019 e un figlio nel 2021. Quello della maternità e dei diritti delle neo mamme sul posto di lavoro sono temi che le stanno particolarmente a cuore e per cui si è sempre battuta, così come il diritto all'aborto. In prima persona si è resa conto delle difficoltà a cui una donna va incontro, quando si tratta di conciliare la propria professione con la vita familiare. Questo a causa dell'assenza di adeguate misure che le tutelino su entrambi i fronti, consentendo di viverli senza colpe e senza rinunce. Sin dalla prima gravidanza, la Creasy ha portato avanti una campagna per migliorare i diritti di maternità per i parlamentari. È stata la prima a nominare un locum, una sorta di deputato provvisorio che potesse ufficialmente e temporaneamente fare le sue veci nella gestione del lavoro (Kizzy Gardiner). Non si era mai visto prima, nel Parlamento britannico. Questa figura, però, le è stata negata una volta data la luce alla sua primogenita. Quando ha annunciato la seconda gravidanza ha contestato le iniziative del governo per limitare i nuovi piani per il congedo di maternità parlamentare. Difatti lei lo ha richiesto alle stesse condizioni del procuratore generale Suella Braverman, massima autorità legale del governo di Boris Johnson, ritenendo di avere gli stessi diritti in quanto donna, al di là dalla carica. Purtroppo in materia ci sono leggi molto restrittive, frutto di antichi retaggi culturali e convenzioni sessiste duri a morire, contro cui la Creasy si batte giorno dopo giorno. Queste discriminazioni mettono le donne in condizione di dover scegliere tra lavoro e famiglia, non le fa sentire libere e autonome nella gestione di questi due aspetti, che potrebbero convivere se ci fossero delle tutele, dei sostegni.Portare suo figlio con sé in Parlamento è stato il gesto simbolico per rivendicare nuovamente la sua battaglia. Lo ha portato per dimostrare quanto sia difficile, quanto impegno serva da parte delle donne, che possono e vogliono farcela, ma hanno bisogno di non sentirsi sole, hanno bisogno di sapere di avere degli alleati accanto, piuttosto che nemici. Rivolgendosi a Jacob Rees-Mogg, leader della Camera dei Comuni di Westminster (padre di sei figli e dunque ferrato sull'argomento) la Creasy ha detto: "Vuole incontrare una delegazione interpartitica per vedere come possiamo garantire a tutti in questo Parlamento la legge sulla maternità e sul congedo parentale?". Lui non si è mostrato particolarmente propenso, difendendo le leggi attualmente in vigore, a suo dire sufficienti e ragionevoli.

Summary: La deputata laburista Stella Creasy è entrata in Aula con suo figlio, nato da poco. Il suo gesto simbolico è in linea con la battaglia che la 44enne porta avanti da tempo: più diritti per le neo mamme che vogliono tornare a lavoro. Per farlo c'è bisogno di supporto, per non sentirsi costrette a scegliere tra carriera e famiglia. Conciliarle è possibile, ma c'è bisogno di leggi e tutele adeguate.

fanpage

it

Summarize: A man who shone a laser into the cockpit of police helicopter dazzling the pilot with a light so bright it could have caused it to crash and kill the crew on board has been jailed. Andrew Paul Holden, 47, was branded'stupid and selfish' by police officers after he shone the laser beam at the National Police Air Service (NPAS) helicopter as it flew over Radcliffe in Greater Manchester. Images of the bright green light seen by the officers show the dangerous 'bloom effect' it creates - easily dazzling the crew with potential catastrophic consequences. Andrew Paul Holden has been jailed after he shone a laser pen into the cockpit of a police helicopter. Officers on the ground were guided to Holden's home in Radcliffe, Bury, by the crew on board the helicopter and were arrested him. The helicopter had to abandon the job it had been called out to in Radcliffe because of the incident. Holden pleaded guilty to endangering an aircraft and was sentenced to six months in prison. The incident took place on October 13 last year when the laser was fired into the cockpit. Holden had previously claimed he didn't target the helicopter intentionally. The pilot on the flight, Captain Robert Grainger, said: 'The consequences of targeting an aircraft can be horrendous. 'During this incident I had to ensure the safety of my passengers, the general public and myself, whilst being disorientated by the intense light. Holden was tracked by officers and arrested at home. 'Luckily this incident didn't result in any injuries, but it could quite easily have ended in fatalities. 'Sadly, and frustratingly, this is not a one off event. I hope that the sentence handed down reflects the severity of such foolish and reckless actions and will serve as a deterrent to others.' The helicopter is equipped with technology which makes it able to track individuals and help with search operations. Tactical Flight Officer, Pc Andy Shaw, said: 'As a result of one person's stupid and selfish actions we had to stop our original tasking, meaning the valuable resource of the NPAS helicopter was taken away from supporting officers on the ground. 'Fortunately, on this occasion it didn't greatly impact on the incident we were dealing with or have any significant impact on the public, but we could have been involved in an incident whereby someone's life was in danger. 'Additionally shining a laser into a helicopter could have itself had catastrophic consequences.' A number of people have been convicted of targeting police helicopters with laser pens to dazzle them, creating dangerous risks for those on board. People who are targeted with the lasers can also suffer eye damage. Last October Chris Vowles targeted the National Police Air Service (NPAS) four times with a laser while he was drinking in his garden below the plane with his friends in Birmingham. Holden, 47, was branded'stupid and selfish' for the incident in Radcliffe, Greater Manchester. Police said shining a laser pen into a cockpit can have potentially fatal consequences for the crew. Police tracked Vowles using thermal imaging equipment so he could be arrested. The 23-year-old TK Maxx worker admitted targeting the helicopter and was sentenced to seven years in prison, suspended for two years. And last November 23-year-old William Sprout, from Warrington, was jailed for up to four months after beaming a laser at a police helicopter from the window of his flat. Police said they take the practice of deliberately targeting helicopters with laser pens very seriously. The lasers can create a 'bloom effect', with a large burst of light dazzling pilots who are in danger of losing control. People have lasers shone into their eyes can suffer eye damage. It's not just police helicopters that are targeted with laser pens. In January last year the RAF revealed its planes had been targeted 262 times in five years. Some of the incidents reported by the military included people shining the lasers into the cockpits of planes carrying injured personnel. The planes were targeted as they descended below 5,000ft on their final approach to Birmingham Airport to take soldiers for treatment at the city's state-of-the-art Queen Elizabeth Hospital. RAF officers have warned that the practice plays 'Russian roulette' with the lives of pilots and those on board, while police have warned of the potentially disastrous consequences. Aviation officials issued a plea in December for people to stop shining lasers into the eyes of pilots as they prepare to land, after 100 incidents were recorded at Liverpool John Lennon airport in just two years. There were more reports of pilots being targeted at Liverpool than at London Stansted and Edinburgh. Shining a laser at an aircraft in flight became a criminal offence in 2010, and carries a fine of up to £2,500. Recklessly endangering an aircraft is punishable by up to five years in jail. The pens can be bought for as little as £8

Summary: Andrew Paul Holden shone a laser pen into the cockpit of police helicopter. Light was so bright it could have caused pilot to lose control of the plane. Helicopter crew helped police on ground track Holden to Radcliffe home. The 47-year-old pleaded guilty to endangering an aircraft and was jailed. Police branded him'stupid and selfish' for his actions with the laser pen. Shining a laser pen was made illegal in 2010, punishable with £2,500 fine.

cnn_dailymail

en

Summarize: this data is currently not publicly accessible. Starting in 1996, Alexa Internet has been donating their crawl data to the Internet Archive. Flowing in every day, these data are added to the Wayback Machine after an embargo period. Sorry, the page you requested could not be found. If you are trying to reach a page from a bookmark, the page URL may have changed. Please choose a community from the navigation at the top of this page. If you are looking for an older news story, it may no longer be here. Free archives of stories are maintained on our site for one month. If you still have trouble finding what you need, please contact our webmaster After Tiger Woods infidelity, Elin Nordegren buys Faglaro Island Swedish mansion Hotgossip.se Faglaro Mansion on Faglaro Island, the $2 million hideaway in Sweden bought by duffer Tiger Woods' humiliated wife, Elin Nordegren, 29. Tiger Woods' humiliated wife has bought herself a little Faglaro Island hideaway as reports swirled that she's moved out on her sex-crazed hubby. Elin Nordegren's real estate purchase in Sweden fueled speculation she might take the kids back to her homeland to start a new life far from her transgressing Tiger. "The official story from her family is that she bought the place with her twin sister, Josefin, not with Tiger," said Poppe Linge of HPG, a Swedish news agency. The fetching blond spent a little more than $2 million to buy Faglaro Mansion, Swedish newspapers reported. Nordegren, 29, has not been heard from since the scandal broke. Radaronline.com reported Monday she has moved out of the couple's $2.6 million Windermere, Fla., home and is bunking at another house nearby. Her Swedish getaway is located on some prime Scandinavian real estate - a small, secluded island reachable from Stockholm only by ferryboat. "It belonged to a professional soccer player who wanted privacy," Linge said. "I heard she started looking at it last summer and bought it two weeks ago." That would be around the time Woods plowed his SUV into a fire hydrant outside his Orlando-area mansion - reportedly after Nordegren caught him texting New York party girl Rachel Uchitel. Uchitel, 34, was the first of at seven reported mistresses (and counting) who have surfaced since then. Woods escaped with just facial cuts and a $164 ticket after totaling his truck. He was not tested for drugs or alcohol despite claims from a witness who "stated that the driver had consumed alcohol earlier in the day," according to a Florida Highway Patrol subpoena request. The witness, who was not named in the document, also told troopers that Woods had prescriptions for the sleeping aid Ambien and the painkiller Vicodin. Woods, 33, publicly apologized for his "transgressions" a few days after the wreck and reportedly offered Nordegren millions to keep her from walking out on him and taking the kids. Meanwhile, Woods was reportedly being courted by yet another woman, talk show queen Oprah Winfrey, who has offered the golfer an outlet to tell his version of the tawdry tale - if not a shoulder to cry on. In addition to Uchitel, a New York party girl, Woods has been linked to six other hotties: - Cori Rist, 31, a divorced mom and a former dancer at the Penthouse Executive Club in Manhattan. - Orlando waitress Mindy Lawton, 33, who lived a little over five miles from Woods' home and who told London's News of the World that he "wanted to spank me and loved pulling my hair as we had sex." - Los Angeles-based porn star Holly Sampson, 36, the star of skin flicks like "Cheating Housewives 6." - Jaimee Grubbs, 24, a cocktail waitress and former contestant on VH1's "Tool Academy" who has admitted to an affair with Woods. - Kalika Moquin, a 27-year-old Las Vegas nightclub executive, who denies a relationship with Woods. - Jamie Jungers, 26, a Vegas cocktail waitress who allegedly had an 18-month fling with Woods. ndillon@nydailynews.com

Summary: Tiger Woods' scorned wife may be moving further away than just another house in the neighborhood-she's purchased a $2 million mansion on a Swedish island. "The official story from her family is that she bought the place with her twin sister, Josefin, not with Tiger," a Swedish reporter tells the New York Daily News, although she supposedly started looking last summer, before Tiger's alleged flings hit the fan, and bought two weeks ago. At least two of those flings involved trysts with Tiger at the home he shares with Elin, which has her "crazy angry," a source tells the Chicago Sun-Times. Regardless, another source adds that the two still want to save their marriage-if they can find somewhere to escape the media, perhaps "a private preserve with total security, in like Montana or Idaho."

multi_news

en

Summarize: Finding out which is the highest mountain in the US Arctic may be the last thing on your mind, unless you are an explorer who skis from the tallest peaks around the globe. Ski mountaineer Kit DesLauriers joined forces with glaciologist Matt Nolan to settle a debate of more than 50 years, while testing a new, affordable mapping technique in a steep mountainous region. Their research is published today (23 June) in The Cryosphere, an open access journal of the European Geosciences Union (EGU). At 6190m, Denali is the uncontested highest peak in North America, but beyond the Arctic Circle, a debate remains as to which US mountain can be crowned the tallest. Depending on the scale of the map you look at, the 1950s US Geological Survey's topographic maps of the eastern Alaska Arctic show either Mt Chamberlin or Mt Isto as the highest mountain in the region. "These mountain peaks just happened to be located in the same area as the glaciers we were studying and several of the peaks ended up in our maps," says Nolan, a professor at the University of Alaska Fairbanks and the lead-author of the study. He had been mapping glacier volume change in the Brooks Range using a technique called fodar, which he invented. "Because we were interested in understanding the performance limitations of fodar in steep mountain terrain, it seemed a natural fit to combine this validation testing with settling the debate on which peak was the tallest." Fodar is used to survey and map terrain using airborne photography. It's similar to airborne lidar, which relies on laser equipment mounted on an aircraft to scan the landscape and create 3D maps of the terrain, but is much more affordable. "The core equipment is a modern, professional DSLR camera, a high-quality lens, a survey-grade GPS unit, and some custom electronics to link the camera to the GPS," Nolan explains. "A modern airborne lidar unit that can map steep mountain terrain like the one we studied costs over 500,000 USD and typically requires a twin engine plane and a separate equipment operator. In contrast, the fodar hardware costs under 30,000 USD if bought new (much cheaper if you buy used) and can be operated by the pilot flying in a small single-engine plane." Taking to the skies, Nolan flew his Cessna 170B to map the Brooks Range peaks. Meanwhile, DesLauriers, a professional athlete and the first person to ski down the highest peaks of the 7 continents, was on the ground climbing up and then skiing down Mt Isto and Mt Chamberlin. During the trek, she tracked her position using the same type of GPS unit Nolan used in his plane. "The GPS antenna, mounted to a steel post in my backpack, required a constant unobstructed view of the sky which forced me to find creative ways to adapt my usual ski carrying system while climbing," DesLauriers says. "Instead of a normal rest stop to eat and hydrate, I used the rare moments standing still to note my location and time in a field journal so that Matt could have as much data as possible to compare our measurements. The process made climbing the peaks, which took on average a 10 hr summit push after a multi-day approach, more difficult but also more rewarding." Nolan explains why this challenging expedition was needed: "The general idea is to measure elevations from the air at about the same time someone is measuring them on the ground. These ground control points then get compared to the airborne measurements and the difference between them is a measure of accuracy." With an accuracy of better than 20cm, Nolan and DesLauriers found that Mt Isto is, at 2735.6m, the tallest peak in the US Arctic while Mt Chamberlin (2712.3m) is only the third highest peak. Fodar measurements also showed a third peak, Mt Hubley, surpasses Mt Chamberlin by about 5m, taking up second place in the list of highest US Arctic mountains. Nolan and DesLauriers believe it is plausible that the ranking has changed over time, and may continue to change as summit glaciers dwindle, though not enough to remove Mt Isto from the top. Fodar has helped settled this debate, but the applications of the technique extend far beyond measuring mountain heights. "Though determining peak heights was a fun and useful study, our primary use for fodar is in change detection in the cryosphere [the planet's frozen regions]." In addition to measuring peak heights, they are using the same maps to study how snow and glacier melt will affect the region. Nolan has also used fodar to measure coastal erosion, permafrost melt, landslides, ice jams, and infrastructure degradation, mostly in Alaska. Elsewhere, he has been discussing fodar projects to study landscape and ecological change in the Galapagos, flooding dynamics in desert regions of Botswana, and even earthquake relief in Nepal. "The possibilities are truly unlimited." ### Please mention the name of the publication (The Cryosphere) if reporting on this story and, if reporting online, include a link to the paper (TBA) or to the journal website. Mountaineers and other adventurers now have a more accurate map of which peaks in the U.S. Arctic are the tallest. Though Denali is the uncontested highest peak in North America — with a summit elevation of 20,310 feet (6,190 meters) — there has been a more than 50-year debate over which U.S. mountain can be crowned the tallest beyond the Arctic Circle. U.S. Geological Survey topographic maps from the 1950s show either Mount Chamberlin or Mount Isto as the highest mountain in the eastern Alaska Arctic region. A new mapping technique, known as fodar, has finally settled the debate. At 8,975.1 feet (2,735.6 m), Mount Isto is the tallest peak in the U.S. Arctic, and Mount Chamberlin (at 8,898.6 feet, or 2712.3 m) is only the third highest. [Photos: See the World's Tallest Mountains] Fodar, which uses airborne photography to survey and map terrain, revealed a third tall peak: Mount Hubley. This mountain surpassed Mount Chamberlin by about 16 feet (5 m) of height, claiming second place in the list of highest U.S. Arctic mountains. Glaciologist Matt Nolan, lead-author of the study published today (June 23) in the journal The Cryosphere, had been mapping glacier volume change using the fodar technique, which he invented. "These mountain peaks just happened to be located in the same area as the glaciers we were studying, and several of the peaks ended up in our maps," Nolan, of the University of Alaska Fairbanks, said in a statement. "Because we were interested in understanding the performance limitations of fodar in steep mountain terrain, it seemed a natural fit to combine this validation testing with settling the debate on which peak was the tallest." Fodar is similar to airborne LiDAR (Light Detection And Ranging), which uses aircraft-mounted lasers to scan a landscape and create 3D maps of the terrain, but is a more affordable mapping option, Nolan said. [The University of Alaska has a more detailed explanation of how fodar works.] "The core equipment is a modern, professional DSLR camera; a high-quality lens; a survey-grade GPS unit; and some custom electronics to link the camera to the GPS," Nolan explained. And, he added, fodar can be operated by the pilot flying in a small, single-engine plane. Nolan worked with champion American skier and ski mountaineer Kit DesLauriers to map the peaks from the air and on the ground. While Nolan flew a Cessna 170B and used his fodar technique, DesLauriers was climbing up and skiing down the mountain range. DesLauriers, who was the first person to ski down the Seven Summits, tracked her position using the same GPS unit Nolan had in his plane. "Instead of a normal rest stop to eat and hydrate, I used the rare moments standing still to note my location and time in a field journal so that Matt could have as much data as possible to compare our measurements," DesLauriers said in the statement. "The process made climbing the peaks, which took on average a 10-hour summit push after a multiday approach, more difficult but also more rewarding." The challenging expedition was necessary, Nolan said, as it offered control points on the ground to compare the airborne measurements to, ensuring accuracy. Now that fodar has settled the Arctic peaks' debate, Nolan said the mapping technique can be used for measurements beyond mountain heights. "Though determining peak heights was a fun and useful study, our primary use for fodar is in change detection in the cryosphere [the planet's frozen regions]," Nolan said. The same maps created from fodar to measure peak heights can help scientists understand how snow and glacier melt will affect the region, Nolan said. He has used fodar to measure coastal erosion, permafrost melt, landslides and more. "The possibilities are truly unlimited," he said. Original article on Live Science.

Summary: It's been the subject of a half-century-long debate. But thanks to a new mapping technique, mountain experts have identified the tallest mountain in the US Arctic and uncovered a bit of a surprise as well. Using fodar, a technique he invented to map terrain using airborne photography, glaciologist Matt Nolan created a high-resolution topographic map of the Brooks Range in Alaska, which reveals Mount Isto to be the tallest mountain in the US Arctic, standing 8,975 feet tall, reports Live Science. But though scientists have long thought that Mount Chamberlin, located about 30 miles away, was Isto's biggest competition, Nolan says it actually falls short of another peak. At 8,914 feet, Mount Hubley stands about 16 feet taller than Chamberlin, he explains in Cryosphere. Nolan was actually using fodar to observe glacier volume change when he realized he was flying over the highest mountains in the US Arctic-only scientists couldn't decide which should wear the crown. That's when Nolan decided to put his technology to the test to find out. As he flew over the Brooks Range in a Cessna 170B equipped with a DSLR camera linked to a GPS unit, skier and mountaineer Kit DesLauriers climbed up and skied down the mountains with her own GPS unit, allowing the pair to chart elevations from the air and ground simultaneously to provide a "measure of accuracy." The resulting maps are accurate to about 8 inches, according to a release. Nolan notes his technology is much like lidar but costs $30,000 rather than $500,000 and can measure coastal erosion, glacier melt, and landslides. "The possibilities are truly unlimited," he says.

multi_news

en

Write a title and summarize: Pregnant women, and their fetal offspring, are uniquely susceptible to Zika virus and other microbial pathogens capable of congenital fetal infection. Unavoidable exposure to Zika virus in endemic areas underscores the need for identifying at-risk individuals, and protecting expecting mothers and their fetal offspring against prenatal infection. Here we show that primary Zika virus asymptomatic infection in mice confers protection against re-infection, and that these protective benefits are maintained during pregnancy. Zika virus recovery was sharply reduced in maternal tissues and amongst fetal concepti after prenatal challenge in mothers with resolved subclinical infection prior to pregnancy compared with mice undergoing primary prenatal infection. These benefits coincide with expanded accumulation of viral-specific antibodies in maternal serum and fetal tissues that protect against infection by the identical or heterologous Zika virus genotype strains. Thus, preconceptual infection primes Zika virus-specific antibodies that confer cross-genotype protection against re-infection during pregnancy. The ongoing Zika virus (ZIKV) epidemic has triggered an explosion in cases of fetal death, microcephaly and other birth defects in surviving infants with congenital infection [1–4]. These sequelae usually occur in parallel with higher and more prolonged maternal viremia for up to 15 weeks following prenatal infection [5–9]. By contrast, ZIKV infection in non-pregnant individuals is mostly subclinical or asymptomatic, and associated with only transient self-resolving viremia [10]. For example, only 19% of ZIKV IgM seropositive individuals reported clinical symptoms during the 2007 Yap Island outbreak [11]. Likewise, only 11% of individuals developed reportable symptoms despite an approximate 50% rate of newly acquired seroprevalance during the 2013–2014 French Polynesian outbreak [12]. Thus, considering ZIKV-associated morbidity is largely confined to infection during pregnancy, there is an urgent need for improved strategies for protecting against congenital fetal invasion and prenatal infection susceptibility. This urgency persists even though several ZIKV candidate vaccines have recently been shown to confer very promising protection in animal infection models [13–21], since clinical validation of efficacy and safety, especially with increasingly recognized immunological shifts that physiologically occur during human allogeneic pregnancies, have not been demonstrated. Given the limited and non-specific clinical symptoms associated with most ZIKV infections in healthy non-pregnant individuals, a fundamental unanswered question regarding prenatal ZIKV susceptibility is whether preconceptual infection protects against re-infection during pregnancy. For some classical prenatal pathogens (e. g. varicella virus, rubella virus), maternal susceptibility and congenital fetal invasion are each efficiently overturned amongst women with resolved infection prior to pregnancy [22–24]. By contrast, protection conferred by preconceptual infection is considerably less reliable for other prenatal pathogens—that may reflect susceptibility to re-infection by immunologically discordant strains (e. g. human cytomegalovirus, influenza virus) [25,26], or attenuated responsiveness of normally protective maternal immune components simultaneously required for sustaining fetal tolerance (e. g. Plasmodium spp., Listeria monocytogenes) [27–29]. Importantly, while prior infection has been shown to protect against lethal re-challenge in non-pregnant hosts for other flaviviruses such as West Nile virus (WNV) or St. Louis encephalitis virus [30–32], whether primary infection protects against re-infection during pregnancy for flaviviruses as a group remains poorly defined since prenatal infection has not traditionally been a dominant feature for these viral pathogens until the recent clinical emergence of ZIKV [33]. Herein, pregnancies established among genetically discordant strains of inbred mice were used to investigate how primary preconceptual ZIKV infection impacts the susceptibility of mothers and their fetal offspring to re-infection during pregnancy. We show that anti-viral immunity primed by asymptomatic infection prior to pregnancy protects against re-infection during pregnancy, with sharply reduced susceptibility in both maternal and fetal tissues. These findings have important translational implications for discriminating individuals at risk for severe prenatal infection from others with naturally acquired protective immunity, and new strategies for protecting against congenital fetal invasion. ZIKV shares with other flaviviruses, susceptibility to innate anti-viral immunity activated by type I interferons (IFNs), and functionally overriding host cell type I IFN responsiveness is required for productive symptomatic infection [34,35]. We exploited the selective resistance of murine cells to STAT2 degradation by ZIKV NS5 protein, reasoning that the natural innate resistance from unabated type I IFN responsiveness in mice makes this species ideally suited to probe immunological shifts primed by asymptomatic infection in non-pregnant individuals [36,37]. In turn, temporally overriding innate protection by initiating type I IFN receptor antibody blockade immediately prior to secondary challenge creates a unique opportunity for investigating the impacts of prior asymptomatic infection on susceptibility to re-infection. Consistent with the results of recent studies [34,35], standard laboratory C57BL/6 mice inoculated with a ZIKV clinical isolate (PRVABC59) from the contemporary Latin American outbreak showed no clinical symptoms and rapidly cleared the infection (Fig 1A and 1B). Comparatively, clinical signs of infection (i. e., hunched posture, ruffled hair, lethargy) that paralleled high levels of circulating virus were unleashed by type I IFN receptor antibody (MAR1-5A3) blockade administered beginning one day prior to infection (Fig 1A and 1B). Thus, antibody mediated type I IFN receptor blockade efficiently causes clinical and virological susceptibility to ZIKV infection despite previously described normal weight gain in mice [34]. To investigate how prior infection impacts susceptibility to re-infection, clinical symptoms and viral RNA levels after secondary ZIKV challenge amongst mice with prior asymptomatic infection were compared with primary infection in naive control animals. Interestingly, despite susceptibility conferred by type I IFN receptor blockade initiated one-day prior to infection, clinical symptoms were sharply attenuated amongst mice with prior asymptomatic infection compared with naive control mice (Fig 1C). These protective benefits paralleled significantly reduced recovery of ZIKV RNA in the blood and tissues of mice undergoing secondary challenge compared with primary infection in naive controls (Fig 1D). Together with recent studies demonstrating significantly reduced susceptibility to secondary compared with primary ZIKV infection in non-human primates [38–40], these results highlight the immunogenicity of ZIKV where even abortive primary infection can efficiently prime protection against re-infection. A variety of adaptive immune components stimulated by ZIKV infection or candidate vaccine formulations are associated with protection. For example, polyfunctional IFN-γ plus TNF-α producing CD8+ T cells with broad ZIKV specificity primed by primary infection can overturn the susceptibility of naive recipient mice after adoptive transfer in vivo [41,42]. Likewise, sterilizing immunity induced by live attenuated or inactivated ZIKV candidate vaccine formulations occurs in parallel with sharply expanded accumulation of IFN-γ producing ZIKV-specific CD4+ and CD4+ T cells [14,15]. On the other hand, the same live attenuated viral strains and nucleic acid based candidate vaccines each prime high titer ZIKV-specific IgG antibodies that protect against infection in non-pregnant mice and non-human primates [13–18]. ZIKV susceptibility to antibodies is further highlighted by protection against infection in pregnant and non-pregnant mice adoptively transferred individual clones of human monoclonal antibodies that bind and neutralize ZIKV in vitro infectivity most efficiently [43]. Thus, while ZIKV-specific T cells and antibodies are each capable of protection, the adaptive immune components stimulated by subclinical infection that protect against re-infection remain undefined. Accordingly, our model of protective immunity primed by primary ZIKV asymptomatic infection was used to evaluate the relative contribution of serological compared with cellular adaptive immune components. Consistent with efficient serological conversion after human subclinical infection [11,12,44], the serum of mice three weeks after asymptomatic primary infection showed robust accumulation of ZIKV-specific IgG antibody, whereas IgA and IgM titers remained at background levels found in naive control mice (Fig 2A). ZIKV-specific IgG antibodies were highly enriched for IgG2a and IgG2b subclasses, with specificity to ZIKV envelope (ENV) and NS1 proteins (Fig 2B and 2C). In turn, serum from mice three weeks after primary asymptomatic infection also efficiently neutralized ZIKV plaque formation in Vero cell monolayers, with serial dilutions of the serum showing reductions in functional activity that directly paralleled when the titer of virus-specific IgG antibody returned to background levels (loss of activity for both after 103 to 104-fold dilutions) (Fig 3A compared with Fig 2A). Interestingly, heat-inactivation did not significantly impact the neutralization potency of serum from mice with prior asymptomatic infection suggesting heat-liable serum components (e. g. complement) are not required for neutralizing ZIKV in vitro infectivity (S1A Fig). To further investigate whether ZIKV-specific antibodies primed by asymptomatic primary infection protect against infection, the susceptibility of naive mice adoptively transferred serum from donor mice infected with ZIKV three weeks prior was evaluated. ZIKV RNA levels were significantly reduced in the serum, spleen, liver and brain of recipient mice prophylactically treated one day prior to infection with serum from donor mice with resolved primary asymptomatic infection compared with the serum of naive control mice (Fig 3B). Interestingly, Fc-γ receptor (CD16/CD32) in vivo blockade [45,46] efficiently overturned the protective benefits of adoptively transferred serum from mice with resolved primary infection, highlighting essential roles for phagocytic host cells that take up antibody coated virus through Fc-γ receptors (Fig 3C). In contrast, depletion of CD8+ or CD4+ T cells, either individually or in combination, did not significantly alter protection against re-infection amongst mice with prior infection (S1B Fig). Thus, protection against ZIKV re-infection primed by primary infection is associated with retained accumulation of high titer viral-specific antibodies that can adoptively transfer protection to naive recipients. Together with the protective benefits of exogenously administered ZIKV-specific IgG antibodies shown in recent studies [43,47], these findings suggest high titer ZIKV antibodies that inhibit viral infectivity can bypass the necessity for CD8+ T cells during primary infection [41,48]. Given the sharply increased morbidity associated with ZIKV infection during pregnancy—with ensuing congenital invasion of fetal tissues, we further investigated whether protection primed by preconceptual infection persists during pregnancy. Three weeks after primary asymptomatic infection, allogeneic pregnancies were established amongst C57BL/6 (H-2b) female mice by mating with Balb/c (H-2d) males to recapitulate the natural heterogeneity between maternal and fetal antigens in outbred populations. Type I IFN receptor blockade was subsequently initiated in pregnant females at midgestation (E10. 5), followed by ZIKV infection one-day later (E11. 5) (Fig 4A). Interestingly, despite immunological shifts required for averting fetal rejection, the presence of expanded fetal target tissue and diminished responsiveness of maternal T cells to primary ZIKV infection during pregnancy [49], protection against secondary challenge was maintained amongst mice with prior preconceptual infection shown by sharply reduced ZIKV RNA levels in their serum, spleen, liver and brain compared with primary prenatal infection in naive control mice (Fig 4A). Importantly, congenital invasion was also efficiently averted as ZIKV RNA levels were reduced to near or below the limits of detection for most concepti (fetal plus decidual tissue) after prenatal challenge of mice with prior preconceptual infection (Fig 4A). Protection against re-infection during pregnancy paralleled persistence of high titer circulating ZIKV IgG antibodies that efficiently neutralized ZIKV plaque formation in vitro (loss of activity for both after 103 to 104-fold dilutions) following preconceptual primary infection compared with naive control mice (Fig 4B and 4C). Interestingly, despite high levels of neutralizing antibodies in maternal serum following preconceptual primary infection, sporadic, low level placental ZIKV dissemination occurred with secondary prenatal infection (Fig 4A). These findings are consistent with breakthrough congenital invasion in mice receiving preconceptual vaccination or exogenous ZIKV-antibody transfer prior to prenatal infection [20,21,43]. To investigate the possibility that vertically transferred maternal antibodies may provide additional protective benefits, the levels of ZIKV-specific antibodies were compared with viral RNA levels amongst individual concepti. ZIKV-specific IgG levels were sharply increased in nearly all concepti homogenates of pregnant mice with prior preconceptual infection compared with the concepti of naive control mice, with a highly significant inverse correlation with ZIKV RNA levels amongst individual concepti scattered across multiple litters with low level breakthrough infection (Fig 4D). Likewise, ZIKV infectivity of Vero cells was significantly reduced by pre-incubation with UV-inactivated fetal tissue homogenates of pregnant mice with prior preconceptual infection compared with the concepti of naive control mice (Fig 4E). Together with recent studies demonstrating protection against congenital ZIKV transmission primed by mRNA or live attenuated viral vaccine platforms [20,21], or amongst mice exogenously administered ZIKV human monoclonal antibody with high neutralization potency [43], these findings highlight the protective capacity of virus-specific neutralizing antibodies in overturning the natural vulnerability of mothers and their fetal offspring to ZIKV prenatal infection. ZIKV is believed to have originated in East Africa, with subsequent mutation into unique West African and Asian variants [50]. Despite incomplete information on whether selective pressures by host immunity drive these antigenic shifts, the existence of unique ZIKV lineage strains has important practical implications for the scope of protection primed by natural infection or vaccination. For example, while purified ZIKV-specific human monoclonal antibodies can effectively neutralize both Asian and African strains, a single amino acid mutation in ZIKV ENV protein can override neutralization by individual antibody clones [43]. Furthermore, while antibodies primed by infection with the related flavivirus, Dengue (DENV), protect against secondary infection by identical viral serotype strains, they can also enhance susceptibility to re-infection by discordant DENV serotypes [51–53]. To investigate whether protective immunity primed by primary ZIKV infection extends to cross-lineage ZIKV strains, susceptibility to the original Uganda East African MR766 strain primed by prior infection with the Asian lineage PRVABC59 strain used in our preceding experiments was evaluated. We found the serum from mice with prior PRVABC59 infection efficiently neutralized plaque formation by the cross-lineage MR766 strain, and with near identical potency compared with monolayers infected with the homologous PRVABC59 virus (loss of activity between 103 to 104-fold dilutions) (Fig 5A compared with Fig 3A). In turn, the potency of MR766 neutralization by the serum of mice with prior PRVABC59 infection was not significantly impacted by pregnancy (Fig 5A). In agreement with this cross-lineage susceptibility of MR766 to antibodies primed by prior PRVABC59 infection, ZIKV RNA levels were significantly reduced after secondary MR766 challenge amongst mice with prior asymptomatic PRVABC59 infection compared with primary MR766 ZIKV infection in naive control mice (Fig 5B). Importantly, cross-lineage protection primed by preconceptual prior infection is maintained during pregnancy shown by significantly reduced ZIKV RNA in the maternal serum spleen, liver, and brain, and amongst individual concepti (fetal plus decidual tissue) after MR766 secondary challenge in midgestation pregnant mice with preconceptual PRVABC59 infection compared with primary MR766 prenatal infection in naive control mice (Fig 5C). Significantly increased ZIKV IgG antibody titers that were inversely associated with ZIKV RNA levels were found in the tissue homogenate of nearly all concepti recovered from protected pregnant mice with preconceptual primary infection, but absent in concepti from naive control mice (Fig 5D). Thus, the broadly neutralizing capacity of serum after primary ZIKV infection in humans, non-human primates and mice [38,40,54], extends to cross-genotype protection against re-infection during pregnancy. Despite identification nearly 70 years ago, ZIKV has remained a relatively obscure human pathogen until its emergence and global spread beginning in 2015. The unique propensity for congenital fetal invasion makes ZIKV distinct from other flaviviruses, and opens-up many unanswered fundamental questions for ZIKV and other pathogens that cause prenatal infection. These include whether strategies that protect against infection in non-pregnant healthy individuals remain effective despite pregnancy-associated anatomical changes that significantly expands susceptible target tissue to include fetal tissues, and increasingly recognized immunological shifts that avert maternal-fetal immunological conflict and dampen the proliferation-activation of T cells after infection during pregnancy [49,55,56]. Here we show preconceptual asymptomatic ZIKV primary infection protects against re-infection, and that these protective benefits are maintained during pregnancy (Fig 6). Thus, naturally acquired immunity against prenatal ZIKV infection is similar to the resistance of mothers to classical prenatal pathogens (e. g. varicella virus, rubella virus) that is also efficiently primed by preconceptual infection. A potentially unifying theme among these pathogens is diversity of protective epitopes that functionally minimizes the impacts of antigenic shifts amongst individual immune dominant microbe expressed antigens. For naturally acquired immunity to ZIKV, this notion is supported by the wide distribution of protective epitopes across spatially distinct domains of immune dominant envelope protein [13–18,43,57–59]. By contrast, the number of protective immune dominant epitopes is more limited for other viral pathogens (e. g. human cytomegalovirus, influenza virus) where protection after preconceptual infection is less reliable [25,26]. These protective benefits conferred by pre-conceptual asymptomatic infection parallel sustained accumulation of ZIKV-specific antibodies in maternal serum that efficiently neutralize virus infectivity in vitro and reduce susceptibility to in vivo infection by the identical or heterologous ZIKV genotype strains. Interestingly, serum from mice primed by pre-conceptual infection efficiently neutralized ZIKV infectivity despite prior heat-inactivation, suggesting these protective benefits do not require complement or other heat-liable components associated with enhanced protection for other flaviviruses such as WNV [60]. On the other hand, antibody-mediated Fcγ receptor blockade overturned protection conferred by transfer of serum from mice with prior preconceptual infection. These results are consistent with our finding that primary asymptomatic infection selectively primes IgG2a and IgG2b antibodies with ZIKV ENV and NS1 specificity, and high affinity Fcγ receptor binding for these specific mouse antibody isotypes [61,62]. Interestingly however, inactivation of Fcγ receptor binding for a human monoclonal IgG1 antibody with high affinity for ZIKV ENV protein does not significantly impact protection against prenatal ZIKV infection in mice [43]. Thus, further studies are needed to investigate how the necessity for Fcγ receptor can be functionally bypassed, in particular focusing on the importance of antibody isotype and/or antigen affinity, and the potential for cross-reactivity of ZIKV-specific antibodies with structurally homologous flaviviruses such as DENV that may promote antibody-dependent enhanced infectivity [63,64]. Complexities inherent to investigating immunity and the pathogenesis of prenatal infection largely stem from the choice of experimental models that need to balance practicality with relevance to human pregnancy. Here, it is important to highlight that only humans have human placentas—with anatomical and molecular features that are not reproduced in any other species [65]. Likewise, the physiological discordance between maternal and fetal-expressed paternal antigens drive potent immunological shifts during pregnancy amongst humans and other outbred species that convey profound impacts on prenatal infection susceptibility [66]. For example, expanded accumulation of immune suppressive regulatory CD4+ T cells is highly accentuated in allogeneic compared with syngeneic pregnancies amongst inbred strains of mice [67]. In turn, expanded systemic accumulation of immune suppressive regulatory CD4+ T cells promotes maternal susceptibility to common prenatal pathogens (e. g. Listeria and Salmonella spp.), whereas dampened suppressive function of maternal regulatory CD4+ T cells fractures fetal tolerance and promotes congenital invasion in the context of allogeneic pregnancy [67–69]. Therefore, to extend the analysis of ZIKV prenatal infection that in mice has been limited to syngeneic pregnancies, MHC haplotype discordant strains of inbred mice were used for breeding to recapitulate the physiological mis-match between maternal-fetal antigens encountered in human pregnancy. Strategies that render mice innately susceptible to more prolonged and higher levels of ZIKV viremia by administration of type I IFN receptor blocking antibodies were exploited to overcome the natural resistance of murine STAT2 to degradation by ZIKV NS5 protein [34–37]. We further reasoned that unabated type I IFN responsiveness that attenuates ZIKV replication and protects against symptomatic disease makes this species ideally suited to investigate the immune response primed by preconceptual infection. Using this model where susceptibility to ZIKV can be temporally controlled by delayed administration of type I IFN receptor blocking antibody, we show even asymptomatic primary infection protects against re-infection, and that these protective benefits extend to re-infection during allogenic pregnancy. Importantly, ZIKV primary infection in wildtype mice used to model human infections that are mostly subclinical and primarily associated with only transient self-resolving viremia does not replicate all aspects of human infection. For example, ZIKV infection in non-pregnant individuals remains asymptomatic despite presumed functional neutralization of type I IFN responsiveness though STAT2 inactivation [10,11]. Thus, the enhanced vulnerability of type I IFN receptor deficient mice that almost uniformly develop symptomatic and often fatal infection [34,70], suggests STAT2-indepenent, type I IFN-dependent cell activation pathways may also be important in protection against ZIKV symptomatic infection. Potential candidates are type I IFN induced STAT3 activation that overrides the pro-inflammatory effects of activated STAT1 and STAT2 in myeloid cells [71], or STAT5 phosphorylation that drives CD4+ T cell differentiation into FOXP3+ regulatory cells [72]. Nonetheless, despite this potential limitation regarding how asymptomatic primary infection is achieved, protective immunity efficiently primed by abortive infection we demonstrate in wildtype mice further highlights the immunogenicity of endogenous viral antigens recently shown with minimally-replicating live-attenuated viral strains, non-replicating inactivated virus or nucleic acid-based candidate vaccine formulations [13–18,20,21]. In the broader epidemiological context, antibody-mediated protection against re-infection during pregnancy primed by preconceptual ZIKV primary infection has important translational implications for new therapeutic strategies aimed at identifying at-risk individuals, and protecting expecting mothers and their fetal offspring. Considering an estimated attack rate that exceeds 90% [44], together with an ~80% rate of subclinical infection among individuals with newly acquired infection [11,12], a majority of reproductive age women in ZIKV endemic areas likely have naturally acquired immunity against re-infection primed by resolved prior infections. Thus, despite very promising protective benefits having been recently shown for several ZIKV candidate vaccines in preclinical infection models involving non-pregnant animals or mice during syngeneic pregnancies [13–21], neither the safety of these formulations, nor their protective efficacy during pregnancies that recapitulate the heterogeneity between maternal and fetal expressed antigens representative of naturally outbred human populations have been established. Thus, our data highlighting that protection against re-infection primed by preconceptual infection are functionally retained during allogeneic pregnancy adds an important, but previously unaddressed perspective on how protection against prenatal infection can be achieved. Our finding that primary abortive infection primes robust expansion of anti-ZIKV neutralizing antibodies that functionally persist during pregnancy and can be found amongst individual concepti proportional to their degree of protection, together with recent studies showing donor serum or purified human monoclonal antibodies can transfer protection against ZIKV infection in pregnant hosts [43,47], points to serological screening for viral neutralizing antibodies as a practical approach for distinguishing susceptible at-risk individuals from those with naturally acquired immunity. Considering reduced placental transfer of maternal antibodies in mice compared with other mammalian species [73,74], the 102 to 104-fold reduced ZIKV levels we demonstrate in individual mouse concepti likely underestimates the degree of protection achievable for human fetal offspring. In turn, the inverse correlation between ZIKV RNA levels amongst individual concepti following secondary infection and levels of anti-ZIKV IgG antibody suggest individual fetal offspring are near the threshold for minimal amount of passively transferred maternal antibody required for protection against congenital invasion in this model using mice rendered susceptible with type I IFN receptor blockade. Taken together, these results showing cross-lineage immunity against ZIKV prenatal infection conferred by primary abortive infection, together with protection primed by minimally replicating, attenuated ZIKV vaccine candidates [14,20,21], underscores the therapeutic potential of preconceptual strategies that prime accumulation of high titer neutralizing antibodies for broadly protecting susceptible reproductive age women. On the other hand, since primary ZIKV infection during early pregnancy in mice also primes the accumulation of neutralizing antibodies that are presumably similarly protective [49], averting congenital fetal invasion may require increased functional thresholds based on antibody levels and/or affinity to viral expressed antigens. Thus, important next-steps are to further investigate the degree of protection against ZIKV prenatal infection primed by preconceptual primary infection, and whether the reduced virus levels we find in maternal tissues are below the threshold required for pathological fetal infection in animals where gestational length, and placental expression of factors that influence antibody function (e. g. neonatal Fc receptor, complement regulatory protein Crry), are more representative of human pregnancy [73–75]. Experiments involving animals were performed under Cincinnati Children’s Hospital Institutional Animal Care and Use Committee (IACUC) approved protocols (Assurance Number 2013–0170). These protocols strictly adhere to recommendations described in the National Research Council’s “Guide for the Care and Use of Laboratory Animals” and American Veterinary Medical Association' s" Report of the AVMA Panel on Euthanasia”. C57BL/6 (H-2b) and Balb/c (H-2d) mice were purchased from the National Cancer Institute Charles River Laboratories (Frederick, Maryland), and maintained under specific-pathogen free conditions at the Cincinnati Children’s Hospital. For all Zika challenge studies, 6–8 week old, sex-matched C57BL/6 mice were randomly assigned to experimental groups. For experiments during gestation, 6–8 week old Balb/c males were used to sire allogeneic pregnancies in C57BL/6 females. Zika virus (ZIKV) strains PRVABC59 (Puerto Rico, 2015) and MR766 (Uganda, 1947) were obtained from the US Center for Disease and Prevention (Atlanta, Georgia) and American Type Culture Collection (Manassas, Virginia), respectively [50]. Virus stocks were propagated and titred based on the number of plaque forming units (PFUs) in semi-confluent monolayers of Vero cells (American Type Culture Collection; Manassas, Virginia). All experiments were carried out under biosafety level 2 (BSL2) containment at Cincinnati Children’s Hospital. For abortive infections, mice were inoculated subcutaneously in the lateral flank with 106 PFU ZIKV suspended in 100 μL sterile saline. For challenge studies, non-pregnant or allogeneic pregnant females (embryonic day 10. 5) with and without primary abortive infection were administered 1 mg anti-type I IFN receptor antibody (MAR1-5A3; BE0241; BioXcell, West Lebanon, New Hampshire) intraperitoneally 24 hours prior to and on the day of infection, and were subsequently boosted (0. 5 mg/dose) every five days thereafter. Mice were checked daily and assigned the following clinical disease score (1 healthy; 2 limited ruffled fur; 3 ruffled fur throughout; 4 mild lethargy; 5 limited movement; 6 moribund or uncontrolled spastic movements; 7 deceased) as previously described [76]. For serum harvest, blood from donor mice was obtained 21 days after initial infection, spun, filtered, and stored at -20°C. For heat inactivation, serum was incubated at 56°C for 30 minutes. For adoptive transfer, 300 μl serum (representing ~1/3 serum volume per donor animal) was administered i. p. to each recipient mouse one day prior to ZIKV infection. For Fcγ receptor neutralization in vivo, 250 μg of anti-CD16/CD32 antibody (clone 2. 4G2, BioXcell) were administered intraperitoneally to mice day -1,0 and 2 relative to ZIKV infection as described [45,46]. For cell depletion, anti-CD4 (GK1. 5; BE0003-1; BioXcell) and/or anti-CD8 (2. 43; BE0061; BioXcell) antibodies (500 μg/mouse) were administered intraperitoneally to mice as previously described [68] one day prior ZIKV infection. Following ZIKV infection, individual tissues were harvested and homogenized, whereas serum was collected after coagulation and centrifugation. Homogenized tissue and serum samples were extracted with the RNeasy mini kit (Qiagen, Hilden, Germany), and levels of ZIKV RNA evaluated by TaqMan (ThermoFisher, Waltham, Massachusetts) one-step quantitative reverse transcriptase PCR (qRT-PCR) on an ABI 7500 Fast instrument, using a previously described primer/probe set: forward primer, 5’-CCGCTGCCCAACACAAG-3’; reverse primer, 5’-CCACTAACGTTCTTTTGCAGACAT-3’; probe 5’/FAM/AGCCTACCTTGACAAGCAATCAGACACTCAA/NFQ-MGB/-3’ [34,77]. Viral burden for each entire tissue was calculated by interpolation from a standard curve produced using serial 10-fold dilutions of ZIKV, and expressed on a log10 scale as number of viral copies. Blood was collected from infected mice compared with uninfected controls and serum was isolated after coagulation and centrifugation. Each individual concepti (fetus plus placenta) was collected 3 days after prenatal infection from primed or naive pregnant females, homogenized in PBS (1 mL) and stored at -80°C. For ELISA, 96-well plates were coated overnight with purified ZIKV ENV (MBS319787) or NS1 (MBS319788) proteins, blocked with BSA (1%), incubated with serial dilutions of serum (starting at 1: 10 dilution) or clarified fetal tissue homogenates (1: 4 dilution). ZIKV-specific antibodies were probed with biotinylated secondary antibodies including rat anti-mouse IgA (clone 11-44-2; 13-5994-82; ThermoFisher), IgM (clone eB121-15F9; 13-5890-85; ThermoFisher), IgG1 (clone A85-1; 553441; BD Bioscience, San Jose, California), IgG2a (clone R10-15; 553388; BD Bioscience), IgG2b (clone R12-3; 553393; BD Bioscience), IgG3 (clone R40-82; 553401; BD Pharmingen), developed with streptavidin-peroxidase (554066; BD Bioscience) using o-phenylenediamine dihydrochloride as a substrate and reading absorbance at 450 nm (A450). For viral neutralization, serial dilutions of the serum from mice with and without primary abortive infection were pre-incubated with 102 PFU ZIKV PRVABC59 or MR766 for 1 hour at 37°C. To investigate functionally neutralizing antibodies in concepti, clarified fetal homogenate was UV treated for 1 hour to inactivate any residual virus [78], and subsequently incubated with 102 PFU ZIKV PRVABC59 at a 1: 100 dilution for 1 hour at 37°C. Following incubation, protection against plaque formation by each virus-antibody complex was assessed in Vero cell monolayers by first incubation at 37°C for 1 hour, followed by overlaying monolayer cells in each well with methyl cellulose (1%), and enumeration of plaques 72 hours thereafter as described [43]. All data were analyzed using GraphPad Prism software. For viral burden, levels of ZIKV RNA within each individual data set, were analyzed using the non-parametric Mann-Whitney test (two groups) or ANOVA (3 or more experimental groups). Linear regression was performed to determine correlation between ZIKA RNA and ZIKA-specific IgG levels in fetal tissue homogenates. P < 0. 05 was taken as statistical significance.

Title: Preconceptual Zika virus asymptomatic infection protects against secondary prenatal infection Summary: Expecting mothers are uniquely susceptible to Zika virus infection that often spreads to vital tissues of the developing fetus. Since Zika virus infection in healthy non-pregnant individuals is mostly asymptomatic, a large proportion of reproductive age women that live in Zika endemic areas have been previously infected, and cleared the infection prior to pregnancy. Here we show that primary Zika virus asymptomatic infection in mice protects against re-infection, and that these protective benefits are maintained during pregnancy. Protection in this preclinical model is mediated by circulating antibodies found at very high levels amongst individuals with resolved infection that efficiently neutralize virus infectivity. Thus, antibodies against Zika virus, naturally stimulated by prior asymptomatic infection, protect against re-infection during pregnancy, and the presence of these protective antibodies may help discriminate protected individuals from others that remain susceptible to infection. This knowledge, combined with protection primed by promising candidate vaccine formulations that also stimulate production of high level neutralizing antibodies, will help identify reproductive age women at-risk for severe infection consequences and more focused therapeutic strategies for averting infection.

lay_plos

en

Summarize: CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of U.S. patent application Ser. No. 13/910,257, which is a continuation of U.S. patent application Ser. No. 11/807,359, now issued as U.S. Pat. No. 8,489,178, which claims priority on U.S. Provisional Patent Application Ser. No. 60/898,506, filed Jan. 31, 2007, and on U.S. Provisional Patent Application Ser. No. 60/817,623 filed Jun. 29, 2006, and which parent application is also a continuation-in-part of U.S. patent application Ser. No. 11/700,729 filed Jan. 31, 2007, and U.S. patent application Ser. No. 11/478,322, filed on Jun. 29, 2006. The disclosures of each of these applications are incorporated herein by reference. FIELD OF INVENTION [0002] In these applications, technology, embodiments and ergonomics of a miniature vein enhancement system is described that uses scanned lasers and other light sources to accomplish the acquisition and projection of a vein pattern to aid the user in locating a vein for venipuncture. In this application, additional capabilities and how these capabilities improve the processes associated with venipuncture are set forth. [0003] Key among these is the use of enhanced embodiments that allow additional data collection and application processes that are critical to the successful performance of venipuncture and related medical and business procedures. BACKGROUND OF THE INVENTION [0004] It is known in the art to use an apparatus to enhance the visual appearance of the veins and arteries in a patient to facilitate insertion of needles into those veins and arteries. Such a system is described in U.S. Pat. Nos. 5,969,754 and 6,556,858 incorporated herein by reference as well as a paper by Zeman, H. D. et al., “The Clinical Evaluation of Vein Contrast Enhancement”, Department of Biomedical Engineering, University of Tennessee Health Science Center, Memphis, Tenn. 8 Feb. 2007. Luminetx is currently marketing such a device under the name “Veinviewer Imaging System” and other information related thereto is available on their website, www.luminetx.com, which is incorporated herein by reference. [0005] In this application, the use of biometrics to identify the user and/or the patient to aid in the management and safety of processes involving venipuncture are described. One such biometric that is unique to this field is to use vein patterns to identify the individual. For example, U.S. Pat. No. 5,787,185, provides for “A method of verifying the identity of an individual comprises capturing an image of the subcutaneous vein pattern at a predetermined region of the individual, converting the captured image to a plurality of stored values representative of the intensity of said image at specified relative locations, processing the stored values to produce a second plurality of stored values representative of the image of the vein pattern having enhanced contrast and subjecting the second plurality of stored values to a thresholding process to select those above a predetermined value and storing a set of measurements derived from the selected ones of said second plurality of stored values for comparison with a corresponding set of measurements made on the individual.” [0006] Current embodiments are large, fixed mounted devices. The present invention allows this capability to be integrated into a handheld device as well as to be used as part of the management of the venipuncture process and the handling of delivered drugs and drawn blood. OBJECTS OF THE INVENTION [0007] 1) It is an object of the invention to integrate data collection capabilities such as bar code decoding and biometrics and use those to control the use of the vein scanner [0008] 2) It is another object of the invention to combine the projection of vein patterns with the projection of data such as text and/or icons to enhance the usability and capabilities of the device [0009] 3) It is another object of the invention to project variable data overlaid on the vein pattern. [0010] 4) It is yet another object of the invention to enable payment schemes such that the practitioner can acquire the device at a reduced initial cost and pay per use or pay per user. [0011] 5) It is yet another object of the invention to gather and then project back on the patient additional parameters of the patient such as pulse and temperature. [0012] 6) It is yet another object of the invention to connect the scanner to a host computer to allow data collected to be stored for future uses such as patient history and billing through wired or wireless means. [0013] 7) It is yet another object of the invention to connect to other devices monitoring the patient and to project the data from those monitors back on to the patient. [0014] 8) It is yet another object of the invention to project the data collected by the invention and other devices onto an arbitrary surface so that the practitioner can view the data. [0015] 9) It is yet another object of the invention to allow the practitioner to avoid blood vessels when performing a procedure on a patient. [0016] 10) It is yet another object of this invention to vary the resolution over the working area to allow certain portions of the viewed area to be scanned in higher resolutions than in other areas thereby using the limited bandwidth of the device in an optimized manner. [0017] 11) It is yet another object of the invention to provide additional lighting on the working area to aid the practitioner to see the patient&#39;s body. [0018] 12) It is yet another object of the invention to provide a mechanism for measuring pulse and projecting the pulse rate back on to the body. [0019] 13) It is yet another object of the invention to provide a mechanism for measuring blood oxygen levels and projecting the value back on to the body. [0020] 14) It is yet another object of the invention to provide a mechanism for reading pulse rate without body contact when the target person is participating in exercise. [0021] 15) It is yet another object of the invention to provide the ability for a remote practitioner to perform or assist in the performance of a venipuncture. SUMMARY OF THE INVENTION [0022] In this application a handheld vein detection device is described with capabilities that allow it to enhance the management of venipuncture processes including blood draw, venous or arterial injection or intravenous starts through the use of bar codes and biometrics; as well as embodiments that enhance usability; and the ability to detect and record additional data about the patient such as the location of venous valves, diagnostic data and re-project that data on to the patient&#39;s body. BRIEF DESCRIPTION OF DRAWINGS [0023] FIG. 1 —this flowchart describes the process for taking blood samples using an embodiment of the invention that combines vein location with other data collection including biometrics and bar code scanning. [0024] FIG. 2 —this flowchart describes the process for injecting medicines into a patient using an embodiment of the invention that combines vein location with other data collection including biometrics and bar code scanning. [0025] FIG. 3 —this exhibit shows the types of bar codes that might be used in the invention [0026] FIG. 4 —this drawing shows examples of text and icons that might be projected by the engine. [0027] FIG. 5 —this drawing shows how variable data can be projected to increase the amount of data available to the practitioner. [0028] FIG. 6A —this drawing shows a body surface being illuminated by a scanning laser pattern. [0029] FIG. 6B —this drawing shows a part of a body being illuminated by a scanning laser pattern, and with portions of the laser light reflected and absorbed off different structures. [0030] FIG. 6C —this drawing shows a contrast in intensity of reflected laser light. [0031] FIG. 7A —this drawing shows how the device can be integrated into a robotic system. [0032] FIG. 7B —this drawing shows the robotic control system of FIG. 7A, with a display and a control system. [0033] FIG. 8 shows a person on a treadmill using the invention. DETAILED DESCRIPTION OF THE INVENTION [0034] Text and Iconography Projection [0035] The preferred embodiment of the invention simultaneously captures information about the target part of the body (e.g., forearm) such as vein location and then re-projects information about the captured image directly over that area of capture. As previously described, in one embodiment a visible representation of the vein position(s) is projected directly on the vein locations thereby highlighting the veins and making them easier for the practitioner to find. [0036] In enhanced embodiments, digital information in the form of text and/or icons can be included in the information that is re-projected on to the skin. The text and icons can be any useful information to assist the practitioner in performing the procedure. This information may be real time information such as vein depth or it may be patient information such as patient name or it may be mistake proofing information such as the quantity of blood needed to be drawn next so that the vial size can be verified. [0037] Referring to FIG. 4, text 1100 is projected that indicates information about the process being performed (e.g., next), the status of the system (e.g., OK) or attribute of the patient (e.g., blood pressure is 110/50). As a further enhancement, icons or pictographs 1101 can be projected that shows information in a graphical manner such that it doesn&#39;t require a common language between users. Further extending the capabilities, combinations of text and icons 1102 can be projected that show both a specific value for a piece of data and a qualitative indication of the importance of the data. In the example 1102, normal temperature is checked, and high and low temperatures are similarly indicated. [0038] An alternative embodiment uses variable markings to indicate some value about the patient&#39;s status. In FIG. 5, an arm 1131 is shown with several veins 1127 / 1132 shown. The projection area 1130 overlaps two veins which are therefore highlighted so that the practitioner can easily find them. Additional information about those veins such as depth can be shown with size indicators 1125 / 1126 / 1128 / 1129. The size of the line between the end marks can be varied to show relative or absolute depth of the vein. If additional colors or intensities are available in the embodiment, the line can be directly overlaid on the vein track 1128 / 1129 in a different color or in a different intensity or both. Alternatively, a negative image in relation to the projection image can be used to show the size bars 1128 / 1129. A further alternative would be to place the size bars in proximity but not on top of the imaged veins 1125 / 1128. [0039] Blood Sample Lifecycle Management [0040] The process of obtaining blood, storing it, transporting it, testing it and returning the test results has many steps, involves multiple persons and is prone to errors. Through the use of the invention many of these mistakes can be eliminated. Referring to FIG. 1, a process for drawing blood samples is described. The process begins 1000 with the verification of the user/practitioner 1001. This verification can take many forms. The device can be equipped with a keypad such that the user can enter in a personal identifier such as employee id-along with an optional pass code. The device can also have a built in token reader such as a smart card reader or an RFID reader such that the user presents this physical token to the device to identify themselves. The device can have built-in biometrics reading capabilities such as a fingerprint reader or an audio input subsystem to allow for voice recognition. Since the device is inherently an image capturing system, image based biometrics such as vein pattern recognition and facial recognition can be used. In most cases a single identifier would be used. However, in some circumstances, a higher level of security might be desired and combinations of identifiers could be used before the process is allowed to continue. [0041] Once an identifier or identifiers have been acquired 1001, they can be compared to a database 1005 of valid users of the device either in a locally stored database or in the case of a scanner that is connected either wired or wirelessly, a remote database can be queried. If the user isn&#39;t found in the database 1003 or if the user does not have appropriate training or privileges for the procedure they can be prompted to try again 1002. This prompt could be in a display built into the device, through audio feedback, through visual feedback of an indicator such as an LED, or in the preferred embodiment, through the projection of text or icons from the scanner&#39;s projection field. If they user has been successfully identified 1006 then the process moves on to verifying that the patient is the correct patient. [0042] Verification of the patient 1007 can take all of the forms described above for the user of the device. In addition, it is common in hospital environments for a patient to wear a bar coded wrist band. The device can be used to scan that wrist band as a means of identifying the patient. Once an identifier or identifiers have been acquired 1010, they can be compared to a database 1011 of valid patients of the device either in a locally stored database or in the case of a scanner that is connected either wired or wirelessly, a remote database can be queried. If the patient isn&#39;t found in the database 1011 or if there are not orders on file for the procedure, the user can be prompted to try again or to correct the situation 1008. This prompt could be in a display built into the device, through audio feedback, through visual feedback of an indicator such as an LED, or in the preferred embodiment, through the projection of text or icons from the scanner&#39;s projection field. [0043] Now that the practitioner and the patient have been validated, a list of blood needed and the equipment needed to perform the procedure 1013 can be presented to the user through one of the modalities described above. [0044] Since the invention can be used for bar code data collection, it can be used to ensure that the appropriate materials are in hand before the venipuncture is performed. As the user selects each piece of equipment 1014 they can use bar code or other means of identification such as RFID to verify that the item 1015 is on the list of needed equipment. The process repeats and prompts the user 1016 to continue with gathering equipment until all of the equipment has been identified 1018. [0045] Once a complete set of equipment has been verified 1019, the user can use the vein scanner to find a vein and perform the venipuncture 1020. As the user fills the vials 1021 they can be identified again as filled and if all of the required vials have not been filled 1023, the user can be prompted to continue 1022. [0046] Once all of the vials are filled 1080 the user verifies that the process has completed successfully 1075 and then packages the specimens taken for delivery to the lab 1027 / 1028 and The status of the completed process is reported either in the documents forwarded on with the specimens and in a preferred embodiment, this information can be updated in to the medical records database using one of the input and connectivity modalities previously described. [0047] The flowchart and the associated descriptions are shown to highlight how additional input and output modalities integrated into the vein scanner can be used to enhance the process of blood collection. However, the specifics of the processes implemented in different medical environments will require changes to the process flow to meet the unique requirements of the specific environment. [0048] Injection Lifecycle Management [0049] Improper or mistaken delivery of medicines is one of the leading causes of accidental injury or death in the healthcare system. In addition to the use of the vein scanner to help identify the position of veins to aid in the taking of blood specimens, the vein scanner can be used to assist in the delivery of medicines into the patient. [0050] In addition, an enhanced scanner can assist in the management of the injection process in a similar way as described above for taking blood specimens. Referring to FIG. 2, a substantially similar process is presented. Steps 1050 - 1069 are handled as previously described for steps 1000 - 1019 in FIG. 1. While blood draw is typically handled with a single catheter and multiple containers, injections might use multiple hypodermics, so step 1070 is not always performed and step 1071 might include multiple injections and therefore multiple uses of the scanner to verify vessel location. All of the remaining steps are substantially identical to that described above. [0051] Intramuscular Injections [0052] A primary focus of the invention has been to locate blood vessels so that a practitioner can access that vessel with a needle for withdrawing blood or injecting a material into the blood stream. In medical practice, some inject-able substances must be injected into the muscle rather than into a vein. Through the use of the invention, veins can be located for this procedure and the area between veins can be targeted by the practitioner thereby reducing the risk of inadvertently injecting the substance into a vein. [0053] Image Capture [0054] In an embodiment of the invention where the laser is scanned in a predictable and repeatable pattern, such as a raster pattern, an image of the targeted area can be captured and stored into memory. This image can be of sufficient quality and resolution to allow image processing techniques that are well known in the art to be applied to this captured image. [0055] As previously described, this stored image can be used for many purposes such as using various image processing techniques to find the vein pattern within the image. In addition to the location of the vein, with appropriate image processing software the scanner can be used to capture and present additional information about objects seen in the field of view. [0056] In a digital camera a photo detector array that has elements that are separately sensitive to red, green and blue light are used with either ambient light or a broad spectrum white light to capture the image. In a laser based system, the image being captured is based on the target surface&#39;s reflection of the light of the specific color (wavelength) of the laser being projected. [0057] For each laser used in the system an additional color can be captured and included in the image. In the preferred embodiment with an infrared and a visible red laser, an image can be captured based on both the reflectivity at infrared and red. Depending on the desired result, one or both of these images can be used. For example, in a bar code application, a bar code could be scanned with the infrared laser that is invisible to the human eye. With the addition of other color lasers, more information about the target surface can be captured. [0058] Finding Needle [0059] Since the needle is a large object that is above the surface of the body prior to venipuncture, image processing software can be used to identify the position of the needle using its different characteristics when compared to skin and sub-skin body parts. Since the invention both captures an image and projects an image, the user can be prompted with projected feedback as to the position of the needle in relation to the nearest vein in the field of view. Since it is important that the needle pierces the vein in its center, icons such as cross hairs or arrows can be used to guide the practitioner to the proper position. For example, right/left/forward arrows can be used to accurately guide the practitioner to the center of the vein by showing them the direction to move the needle. The invention is also able to use other feedback mechanisms such as audible feedback or that of projecting text into the field of view with prompting information. [0060] Finding Vein [0061] Once the image is captured from the penetrating infrared laser, many algorithms well known in the art can be used to find the veins. Several things are typical in the types of veins that one would wish to select for venipuncture, including that it has a sizeable segment that is linear. Therefore, a line detection algorithm can be used. Another thing that is characteristic of the vein image being captured is that there are edge boundaries between the area surrounding the vein and the vein itself. An edge detection algorithm such as the one described by Canny in the IEEE Transactions on Pattern Analysis and Machine Intelligence archive, Volume 8, Issue 6 (November 1986), Pages: 679-698, Year of Publication: 1986, ISSN:0162-8828, can be used to find these edges. [0062] Finding Depth [0063] In a preferred embodiment, the output intensity of the laser (or the gain of the photo detection circuitry, or a combination of both) can be varied from frame to frame as the scanner passes over the target area. The higher the intensity, the deeper the laser will penetrate the body. At a low setting, the veins identified will be close to the surface. At higher settings, more and deeper veins will be identified. [0064] By comparing the images between frames a determination can be made of which veins are close to the surface of the body and which veins are deeper. As the intensity increases and new veins become visible, the image processing software can tag the veins seen with an indicator of the intensity at which they first became visible. This provides an indication of relative depth. [0065] Several user interfaces are possible. In a system with multiple, differently colored projection lasers, veins at different depths can be displayed in different colors. In a system with a single projection color, the lines that represent the veins could have patterns superimposed over the vein to indicate depth such as crosshatching for shallow and solid for deep. An alternative embodiment would be to allow the user to set what depth to look at so that only the veins of a specific depth are presented to the user. [0066] Bar Code [0067] There is a significant body of knowledge on the use of image capture devices to read bar codes such as described U.S. Pat. No. 6,223,988. Since the invention is capable of capturing an image of the necessary resolution and quality for bar code detection, algorithms well known in the art can be used to find and decode bar codes that are in the field of view. [0068] Bar codes are commonly used to identify users (through ID cards or badges), identify patients (through wrist bands) and to identify equipment (through bar-coded labels) in medical environments. It is an important extension of the invention to have it able to read bar codes so that it can be fully integrated into the work flows of the medical environment. [0069] The invention can be implemented such that it captures images of the field of view, and stores those images into computer memory. Using techniques well known in the art, the captured images can be searched for the presence of bar codes such as those shown in FIG. 3. Both the 1D and 2D bar codes shown in FIG. 3 could be captured by an image capturing version of the invention. An alternative version of the invention can use the signal from a single line of the laser scanning pattern, as it crosses a 1D bar code as seen in the upper portion of FIG. 3, and decode the bar code through the reflections of the marks and spaces through techniques that are well known in the art. [0070] By re-using the image capture capabilities of the scanner for bar code reading a very small, tightly integrated, low cost embodiment can be created that combines vein location and bar code scanning. [0071] An alternative implementation of bar code scanning would be to use a commercially available off the shelf bar code scanning engine such as those available from Hand Held Products integrated into the housing of the invention. This would have the advantage of keeping the vein detection portion of the design simpler and forgo the need to develop or integrate bar code reading into the vein detection electronics and software. The disadvantage is that there would be redundancies that might increase the size and cost of the unit. [0072] Biometrics [0073] As previously described, the ability to identify the user of the invention and the patient upon which the invention will be used are very useful to the management of the medical procedures. Simple methods of passwords or PINs can be used for the user identification, and as previously described bar coded wrist bands can be used to identify the patient. However, each of these approaches has its weaknesses in that the password or PINs can be misplaced and the wrist band is only available in a hospital environment. [0074] Once again, the image capture capabilities of the invention can be used for capturing many biometric identifiers such as but not limited to fingerprint, face recognition, iris or retina recognition, and vein pattern recognition through techniques well known in the art. [0075] There are many biometric capture devices available on the market that are designed to be integrated into an OEM devices such as laptops. This includes fingerprint modules such as those available from AuthenTec (www.authentec.com) which can be integrated into the invention. This avoids the complexity of the user having to ‘take a picture’ of their finger and allows them to simply swipe their finger on the sensor. [0076] While fingerprint has become a common method of biometric identification, any biometric identification device could be integrated into the invention to allow the user and patient to be identified. [0077] Non contact biometrics can be valuable in a medical situation. For example, a fingerprint would be difficult to read from the gloved hand of the practitioner and the device would have to be cleaned between patients if they came in physical contact with it for a fingerprint swipe. Therefore, non contact biometrics are very desirable. By adding a microphone and associated analog to digital circuitry, sound sampling can be included in the invention and voice recognition techniques well known in the art can be applied to user and patient identification. [0078] Non-Linear Scanning Speeds [0079] The scanning speed of a moving laser and the resolution of the captured image are subject to many physical limitations such as the maximum speed of the mirror, and the bandwidth of the electronic circuitry. However, it is desirable in some embodiments to have a higher resolution of the image on a particular area of the body than the device is inherently capable of. One mechanism to overcome this limitation would be to reduce the area being scanned so that whatever the native resolution of the device is, it is being applied to a smaller physical area. This trades off scanning area vs. resolution. [0080] An alternative embodiment of the device would is to change the resolution of the scanned image in a particular area of the field of view. For example, since the practitioner would typically focus their attention on the center of the field of view, if the device can be made to have higher resolution at the center, the information content of the projected image would be higher at that most important point, without reducing the total field of view of the device. In a scanned laser implementation, this is done by varying the amplitude of the drive circuit so that the mirror moves less quickly in the center and more quickly outside of the center. The increase in dwell time at the center of the image provides higher resolution in that area. The decrease in dwell time outside the center would reduce the resolution in that area. The advantage of this over the previous embodiment is that the total field of view is maintained while sill gaining an improvement in resolution of the targeted area. [0081] Business Models [0082] Most medical practitioners will have the need to draw blood or otherwise stick the patient in their normal course of practice. However, for many of these practitioners this is not done on a regular basis. For example, in a general practitioners office this might be only two to three times a day. This presents a dilemma. On one hand, they don&#39;t do the procedure often enough to become very skilled, and would therefore gain an extraordinary benefit from a vein location device, but on the other hand they don&#39;t perform the procedure often enough to cost justify the acquisition of such a device. [0083] Records of usage can be: 1. Uploaded immediately through a wireless connection 2. Kept in the device memory and then uploaded to a billing system later through a wired or wireless connection 3. Kept in the device on a removable storage device such as an SD card and then transferred to a billing system by removing the card from the device 4. Entirely within the device by using a pre-paid billing approach where a certain number of usages are enabled and the device counts down until there are none left and then disables itself until such a time as more credit is added, through a wired, wireless or physical token as discussed above. [0088] To overcome-this dilemma, the device can be modified to provide a per use-type licensing model. This can be approached by keeping track of the number of times the device is activated in memory that is built in to the device. Various schemes can be used to activate the device such as: 1. Enable the device for a pre-determined period of time. This can be cheated in that there is no limit to the number of patients scanned during that period of time. 2. Enable the device for an unlimited period of time after a particular patient is identified through a biometric scan such as vein pattern, face or voice recognition. This is more expensive to implement, but is much harder to cheat. 3. Some combination of 1 and 2. The inverse of this dilemma is an office with many practitioners where they share the device. In such a circumstance, the benefit of the device is very large when the entire group is considered. It would desirable to be paid more for the device in such a circumstance. Through the use of biometrics, the practitioner can be identified and a record can be kept in the device of that practitioner&#39;s access of the device. As above, various schemes can be used to activate the device such as: 1. Enable the device for a pre-determined period of time. This can be cheated in that there is no limit to the number of patients scanned during that period of time. 2. Enable the device for an unlimited period of time after a particular patient is identified through a biometric scan such as vein pattern, face or voice recognition. This is more expensive to implement, but is much harder to cheat. 3. Some combination of 1 and 2. [0095] Data Collection [0096] In the simplest embodiment, a vein scanner is a stand alone device that simply detects and projects a vein pattern back on to the skin of the patient. In this embodiment, no data about the process or the patient is retained for future use. In an enhanced embodiment, the device can be used to collect, store, manipulate and transmit information about the user, the usage of the device, and the patient. Also, updated control software and firmware can be uploaded to the device. In addition, data can be transferred into the scanner that can be used as previously described to manage the processes associated with venipuncture. [0097] The transfer of information can either be batched up so that when needed the device can be connected to a computer system for upload and download, but in normal operation it is unconnected, or the device can be always connected to a computer system either through a wired or wireless connection. There are advantages and disadvantages and costs associated with each of these techniques that are well known. [0098] While a bi-directional connection that allows the systems to acknowledge that communications have been properly received, a one-way communication scheme can be used for simple embodiments. [0099] Wired Connectivity [0100] The device can be connected to a computer system using a cable such as commonly seen in bar code scanners at store checkouts. The advantage of this implementation is that it is typically low cost and doesn&#39;t require that a battery be in the handheld and eliminates the need to charge the battery. The disadvantage is that the unit cannot be moved far from the computer and the cable can get in the way and easily breaks. Approaches well known in the art can be used for cabled communication such as RS232, USB or Ethernet can be used for transport medium. [0101] Wireless Connectivity [0102] A wireless implementation eliminates the problems associated with a cable with the tradeoff that a battery is needed in the device. Wireless can be implemented with well understood approaches such as point-to-point proprietary protocols such as seen in remote control key fobs or standard protocols such as Bluetooth or WiFi. [0103] Optical Connectivity [0104] Since the device contains both a means of measuring light with its photo detector and emitting light with its lasers, these devices can also be used to transmit and receive data. Thorough the use of modulation schemes that are well known in the art, the laser can be used to transmit data to a remote receiver that could be integrated into the cradle or into a stand alone receiver. This receiver would have a photo detector system capable of detecting and decoding the modulated laser light. It would also have an output system such as a LED or LEDs that would transmit a modulated light signal at a wavelength that can be detected by the photo detector in the handheld. In this manner a two-way communication session can be established. [0105] Batch Connectivity [0106] Memory can be added to the system so that system events can be captured and system control information can be kept in that memory. From time to time, the information collected will need to be sent to another computer or new control information will need to be downloaded to the scanner. Any of the communication techniques previously described can be used on an ad-hoc basis to connect the scanner to a computer and to transfer the information. When not transferring information, the scanner is disconnected from the computer system and continues to operate in a stand alone mode. [0107] Flashlight [0108] In many field medical situations, there is limited ambient light available for performing procedures such as venipuncture. One mechanism that a practitioner would use to improve lighting would be to use a handheld flashlight to illuminate a specific area of the patient&#39;s body. One embodiment of the invention would be to integrate the scanning capability into a flashlight and the inverse of integrating a flashlight into the device. [0109] One embodiment is that the invention be used such that the intensity of the projected field is bright enough to illuminate the body area without the need of additional lighting during the venipuncture process. This can be implemented so that the intensity is controllable by the user with the same hand that is holding the scanner. By varying the intensity of both the dark areas that represent veins and the light areas that represent the spaces between veins, and keeping the contrast between the two sufficiently different, the field of work can be kept well lit while maintaining the ability to discern the vein positions. [0110] Integrated Scanner into a Flashlight [0111] Most of the embodiments of the invention have been described as stand-alone devices. However, since the scan engine of the invention can be miniaturized, it is uniquely suited to be integrated into other devices already in use by the practitioner. For example, the scan engine can be built into an otoscope such that it becomes a multi-function device for both looking into shaded areas of the body such as the ear canal and then used for vein location. The advantages to the user are that they have a single device to buy and manage and a single battery can be used. [0112] Another device that the invention can be integrated with is a flashlight. Depending on the application, the scanner can be integrated so that the light beam and the scan/projection field are aligned along the same path. In this manner, the area of the body is illuminated by the flashlight and the vein pattern is viewable in the same field. The laser projection will be bright enough for most applications, but the flashlight/scanner combo can contain controls that alternately switch between scan/project only, light only and both or other combinations of those modes. The intensity of both the flash light and the projector can be controlled in these modes to ensure that the pattern remains visible without becoming over-bright or too dim. [0113] Integrated Flashlight into the Scanner [0114] An alternative approach is to add lighting capabilities to the scanner. Finer control over the modes of the device can be accomplished in this way. In a preferred embodiment, the scanner is implemented so that multiple lasers allow any color to be projected on to the body including white. In this way, the field of view can be illuminated in white light and the other colors can be used to identify the position of the veins and to project data into the field of view. Since there is no competing illumination from a flashlight bulb, the image will be easier to read, while still illuminating the body part. [0115] A further enhancement of the invention would be to implement a non-linear scanning pattern so that the illuminated area can be made arbitrarily wide, thereby increasing the device&#39;s utility as a flashlight while maintaining a good vein image in the center of the scan area. [0116] A user interface can be implemented that allows the practitioner to switch between these modes and to modify the parameters such as intensity, field of view and field of high resolution. This user interface can be either controls for user input or based on the distance to the target area or both. [0117] Telemedicine [0118] The use of robots to provide fine control of surgical tools is well known in the art. In U.S. Pat. No. 7,155,316, a robot system for use in surgical procedures is described. In U.S. Pat. No. 7,158,859, a mobile robot that provides the ability to remotely control the robot and to see the patient, the patient surroundings and the tools in use is described. [0119] In a normal patient encounter for venipuncture, the practitioner would use several senses to locate a vein and then to perform venipuncture. This would include looking for both visible and tactile indicators of the vein position. The invention enhances the visible feedback of the position of the veins. By using the visual position enhancement, the ability to use a tele-robot to perform venipuncture is enhanced. [0120] The vein scanner engine can be integrated into one of these robots so that the scanning/projection field is positioned in alignment with the end of a robotic arm which contains the needle or catheter for the venipuncture procedure. The engine can be mounted directly to the arm that contains the needle or it can be mounted to a separately controllable arm. [0121] The engine can work in one of three modes. It can work as it normally does such that the acquisition of the vein pattern and the projection of the vein pattern is directly on the patient&#39;s skin. In this mode, the cameras that are already on the robot can view the pattern on the skin and send that back to the remote operator on the screen that is already provided for the remote operator. [0122] In a second mode, the detected vein pattern is captured and then rather than projecting the pattern back on to the patient, the pattern is sent to the remote practitioner and is shown to them on a screen. In this mode, there are several possible embodiments. One is that the pattern of veins can be overlaid on the video display of the patient&#39;s body. In this manner, what they see on the screen is very much like what they would see on the body. The advantage of this is that the image from the vein scanner can be enhanced and made brighter than it would be in the first mode. [0123] In a third mode, a combination of the first two modes can be used that allows the practitioner to switch between the modes or for both modes to be used simultaneously on two different screens or windows on a single screen. [0124] Referring to FIGS. 7 a and b, a robotic control system 1204 is shown with a controllable articulated arm 1200 holding a medical device 1205. The remote medical practitioner 1207 uses a corresponding control system 1206 to move the arm 1200 and the medical device 1205 to perform a medical procedure on the patient 1201. The remote system 1206 has a display 1208 that the practitioner uses to view the patient 1201 and the action of the medical device 1205. Typically this is performed by a camera mounted to the medical device 1205 or to the arm 1200. Our invention adds a vein scanner device 1202 to either the medical device 1205 or the arm 1200. The vein scanner is arranged in such a way that the scanned area overlaps in a known orientation with the medical device. In a preferred embodiment the medical device is a syringe and the enhanced vein pattern is used to properly position the syringe for venipuncture. The invention enables this by either projecting the pattern on the patient&#39;s body such that it is viewed by the camera and seen by the practitioner on the remote display 1208 or alternatively a second display is mounted along with the camera display 1208 so that a digital version of the vein pattern can be seen or alternatively a single display is used with the digital version of the vein pattern overlaid over the camera view of the patient. [0125] Non Contact Photoplethysmography [0126] Pulse oximetry is a non-invasive method of measuring arterial oxygen saturation. The use of an infrared light source and a red light source along with a photo detector in a finger clip arrangement to measure pulse and oxygen levels in blood are well known in the art. The basis for the measurement is the differential absorption between oxygenated hemoglobin and non-oxygenated hemoglobin of the two different wavelengths of light. The traditional technique for performing this measurement requires contact with the body and relies on the transmission of the light through an area of the body with a small cross section such as a finger or an earlobe. By alternating between the red and infrared light sources and measuring the level of the light that is passed through a body part, a waveform that is known as a photoplethysmograph (PPG) is captured. Through signal processing techniques that are well known in the art, oxygenation levels and pulse rates can be calculated. [0127] Several articles and patents have been published that use non-contact techniques to measure the necessary waveforms by replacing the photo detector with a remote device such as a CMOS camera. U.S. Pat. No. 6,491,639 describes the use of a light sources and the photo detector in the same plane eliminating the need for a small cross section part of the body to be used for measurement of the necessary waveforms. This design relies on the internal reflections of the body such that the light is directed in to the body and then reflected out of the body rather than completely passing through a small cross section portion of the body. [0128] In the article by Humphreys, K. et al, “A CMOS Camera-Based Pulse Oximetry Imaging System.” Proceedings of the 2005 IEEE Engineering in Medicine and Biology 27th Annual Conference (2005). 7 Feb. 2007, they describe a non-contact pulse oximetry system that uses a CMOS camera as the detector. In this design, the detector is held at a distance from the body and a bounding box is selected in the captured image. The average intensity of the bounding box is used as a proxy of the photo detector value that would be found in a contact pulse oximeter. By plotting the changing value of the average intensity of that location over time a PPG is derived that can be used to measure oxygenation and pulse using the same techniques as previously described. [0129] Our micro vein enhancer invention uses scanned lasers to capture a pattern of veins below the surface of the skin and then re-projects a vein pattern image directly back on to the body. In one embodiment those laser light sources are visible red for projecting the image and infrared for detecting the veins. The vein detection relies on absorption characteristics that are related to those used in pulse oximetry in that the infrared light is absorbed differentially by hemoglobin than surrounding tissues. [0130] Furthermore, we have previously described how a micro vein enhancer can be used to capture images based on the light transmitted by the lasers and reflected back to the micro vein enhancer&#39;s photo detector circuitry. Through the combination of these techniques, a miniature vein enhancer can be extended to read pulse and oxygen levels without contact with the patient. [0131] Referring to FIG. 6 a, view of the body surface 1150 is being illuminated by the scanning laser pattern 1160. Since information must be collected separately from each laser wavelength, a mechanism is required for the system to differentiate the reflected energy from each laser. One mechanism is to use optical filters and two photo detectors, each responsive to only one of the lasers. Another mechanism is to polarize the lasers so that two photo detectors, each sensitive to only one of the laser&#39;s polarization. [0132] A further mechanism is to project using both the visible red and infrared lasers in an alternating pattern. The alternation can be handled in multiple ways. Possibilities include the lasers time sliced so that short pulses of each laser are alternated while the laser scans. Alternatively, the lasers can be switched between scan lines with right to left with one laser and left to right with the other laser. Alternately, the lasers can be switched on some multiple of scan lines or on alternate complete passes of the scan area. [0133] Referring to FIG. 6 b, a part of the body 1153 is illuminated by the scanning laser pattern 1154 from the vein enhancer. Portions of the laser light are reflected and absorbed off different structures of the body. The portion that is reflected 1155 is measured by the photo detector of vein enhancer. The laser projects a point source on to the body. Therefore the reflected signal is characteristic of a small area of the body. Since the PPG needs to be representative of both venous and arterial blood, the small point source reflection may be insufficient for detecting the PPG. [0134] Referring back to FIG. 6 a, the scanned area 1160 is broken down in to a number of smaller areas (e.g., 1151 / 1152 ) that are sized to capture a large enough area of the body such that an adequate PPG can be detected. The system can dynamically change the size of the smaller areas such that they are sufficiently large to obtain an adequate return signal regardless of the distance of the scanner from the body. The system then averages the returned signal from all of the pixels in the smaller areas and records the values. By keeping track of these values for the same location, over time, a PPG is generated. [0135] Since the preferred embodiment is a handheld device, the motion of the device in relation to the patient must be allowed for. In the simplest embodiment, since the device is breaking the scan area down into a series of subsections and generating the PPG from the subsections and there would be similar return signals from these subsections, the motion can be ignored. However, if the motion becomes too severe or if the area being imaged is heterogeneous, then the system can use digital or optical image stabilization well known in the art to maintain alignment from frame to frame. An alternative method would be to keep PPGs from all of the subsections, then as the hand moves, the system would use the PPG from areas that still remain in view. [0136] Non Contact Pulse Measurement [0137] Many commercially available exercise machines such as treadmills and bicycles provide the capability to measure pulse rate. Knowledge of the current pulse rate is very valuable for managing excursive intensity. One commonly seen embodiment relies on hand contact with electrodes that are used to measure heart rate. In many situations, such as running, it is difficult or dangerous to maintain contact with the electrodes. Another common embodiment uses a chest strap that measures heart rate and then wirelessly transmits it to the exercise machine so that the value can be displayed. This requires that the user acquire this separate device in order for pulse rate to be displayed. In a preferred embodiment of the invention, a capture only scan engine is mounted in proximity to the user such that the scanning pattern is oriented such that it strikes an uncovered portion of the body such as the hands, arms, neck or face. As described previously, the PPG waveform can be captured and the pulse rate can be fed back into the exercise machine and displayed to the user. [0138] Referring to FIG. 8, a person is shown exercising on a treadmill. A typical treadmill is composed of the bed 1305 which houses a moving belt, an upright 1302 at the front of the treadmill and a combination control panel, display panel and handle at 1303. The invention needs access to bare skin and therefore needs to be mounted in proximity to typically uncovered locations such as the legs 1310, the arms 1309 or the neck 1312 and head 1308. In addition, to normally uncovered body parts during exercise, in some embodiments it would be desirable for the torso 1313 to be bare or for a shirt made from materials that are transparent to the wavelength of the scanning light to be worn. Various embodiments of the invention can have at one or more of locations including a laser scanner engine 1304 mounted on the treadmill upright 1302, or an engine mounted in the display panel 1303. In addition, an additional upright 1307 can be added to the treadmill to allow for a scanner engine 1306 to be mounted above the standard parts of the treadmill. Given that the person will typically be moving, it is preferred that the scanner target a part of the body that moves the least. Therefore, the preferred embodiment would target the upper part of the body rather than the legs or the arms. Given that the pulse measurement application requires much lower resolution than the projector application described elsewhere, the full travel of the moving mirror can be used to steer the measurement zone over a fairly wide range of locations. This agility provides two benefits. One, the beam can be steered to the general desired location and the beam can be moved in synchrony with the movement of the body during exercise. Beam positions 1300, 1311 and 1301 show nominal center points for the beam. However, they can be steered in real time as desired within the general area of the center point.

Summary: A portable handheld vein-image-enhancing device broadly includes: a first laser emitting a beam of light at a first wavelength; a second laser emitting a beam of light at a second wavelength; a scanner to scan the beams of light onto the target surface; a photo detector to receive the reflected vein image and convert the image into a signal, for use by the second laser and scanner to project the image onto the target surface; and a user interface and electronic circuitry to permit adjustments to one or more of the following imaging parameters: a vein size parameter to set a vein size to be imaged by the device; a field of view size; a size for a field of high resolution for the projected image; a brightness of the projected image; and a laser output intensity parameter setting a depth of penetration by the first wavelength for the imaging.

big_patent

en

Summarize: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method of determining the position of a catheter in a patient&#39;s esophagus, a control unit for use with a ventilator, and a computer program product for use in such a control unit. 2. Description of the Prior Art and Related Applications It is known in the art to use myoelectrical or neuroelectrical signals from a patient to control the function of a ventilator providing breathing support to the patient. U.S. Pat. No. 5,820,560 and U.S. Pat. No. 6,588,423 both disclose methods and devices for triggering ventilatory support to a patient using a myoelectrical signal obtained from the diaphragm. U.S. Pat. No. 5,671,752 discloses a method and a device for registering the myoelectrical activity of the diaphragm by means of an esophageal catheter having an array of electrodes. The signals from such a catheter can be used as the myoelectrical signal to control ventilator function. EP 1 091 780 discloses the use of a neuroelectrical signal picked up, for example, from the phrenic nerve to control a ventilator. A problem when obtaining a myoelectrical signal from the diaphragm is positioning of the catheter within the patient&#39;s esophagus. To obtain a proper signal some of the electrodes of the catheter should be placed above the diaphragm and some below it. There is a possibility that the catheter will be inserted too far, or not be inserted far enough. In both cases, the catheter will detect a weak signal, or may not capture any signal at all. Alternatively, the catheter may capture myoelectrical signals from other muscles instead of, or in addition to the signal from the diaphragm. Hence, it is difficult to obtain an optimal catheter position and the ventilator may have to work in pneumatic triggering mode if the signal is too weak. There are some problems associated with methods based on the registration of the EMG signal from the diaphragm. There may not be an EMG signal present, for example if the patient is sedated or has no breathing activity of his own for other reasons. The EMG signal may be very weak and/or difficult to detect, for example because of disturbances caused by breathing support provided to the patient. There is a risk that other myoelectric signals resembling that of the diaphragm but originating from other muscles are mistaken for signals from the diaphragm. Such signals may come, for example, from the abdominal muscles. Co-pending application No PCT/EP2007/054149 discloses a method of positioning the catheter based on the ECG component that will always be present in a myoelectrical signal from the diaphragm. In this application, the damping of the ECG signal caused by the diaphragm is used. The ECG signal components from different electrode pairs are determined and compared and the difference in amplitude of the ECG signal between different electrode pairs is used to determine the position of the diaphragm relative to the electrode pairs. The greatest damping between two neighbouring electrode pairs should be caused by the diaphragm being positioned between these two electrode pairs. This method is predominantly based on the registration and comparison of the QRS complex of the ECG signal. Co-pending Swedish patent application No. 0850076-1 discloses a method utilizing the presence of the p wave of the ECG signal. Since the damping of the p wave is very strong with increasing distance from the heart, any electrode pair picking up a p wave must be located fairly close to the heart&#39;s atria, well above the diaphragm. SUMMARY OF THE INVENTION It is an object of the invention to determine the position of an esophageal catheter inserted in a patient, in relation to the patient&#39;s diaphragm. This object is achieved by a method of determining the position of an esophageal catheter inserted into the esophagus of a patient, said catheter having a number of electrodes arranged to pick up a myoelectrical signal, including the following steps: stimulating the muscular activity of the diaphragm at a specific point in time or period, and detecting resulting signals respectively with different pairs of said electrodes supplying the signals detected by the respective electrode pairs of the esophageal catheter at the specific point or period in time to a computerized processor, and in the processor, automatically determining which of said electrode pairs detected a signal with the highest amplitude. The object is also achieved by an apparatus for determining the position of an esophageal catheter inserted into the esophagus of a patient for picking up myoelectric signals from the patient, including at least one stimulating electrode arranged to stimulate the muscular activity of the diaphragm by applying a stimulus signal, a catheter having a number of electrodes that respectively detect a myoelectrical signal resulting from the stimulation of muscular activity by the stimulus signal, a registering unit that registers the signals detected by respective pairs of the electrodes of the esophageal catheter at the specific point in time and the stimulus, a control that operates the stimulating electrode to cause stimulating the electrode to stimulate the muscular activity at a specific point in time, and that automatically determines which of said electrode pairs detected a signal with the highest amplitude at the specific point in time, based on the signals received by the registering unit. The object is also achieved by a non-transitory computer-readable storage medium encoded with programming instructions for controlling an apparatus for determining the position of an esophageal catheter inserted into the esophagus of a patient, the catheter having a number of electrodes arranged to pick up a myoelectrical signal, and at least one electrode arranged to stimulate the muscular activity of the diaphragm, when the storage medium is loaded in a control unit of the apparatus, the programming instructions cause the apparatus to: stimulate the muscular activity of the diaphragm at a specific point in time, register the signals detected by respective pairs of the esophageal catheter at the specific point in time, determine which of the electrode pairs detected the stimulus signal with the highest amplitude. By stimulating the diaphragm and registering the signal at the time of stimulation it is ensured that the registered signal really is the signal from the diaphragm and not another bioelectrical signal that resembles that of the diaphragm. By registering the signals from all electrode pairs of an esophageal catheter at the time of stimulation, the ones registering the strongest signal can be determined. This electrode pair or these electrode pairs will be the ones closest to the diaphragm. In this way the position of the catheter relative to the diaphragm can be determined and adjusted as desired. The stimulus signal may be applied transcutaneously or subcutaneously, using any known method and suitable electrodes for applying the signal in the desired way. The stimulus signal may be applied to a nerve controlling the function of the diaphragm, such as the phrenic nerve or to the diaphragm itself. In a preferred embodiment the control unit determines if the electrode pairs that record the strongest stimulus signal are located in or close to the middle of the catheter and, if they are not, indicate that the catheter should be adjusted. This is preferably achieved by means of a computer program algorithm. This will provide a direct feedback to an operator that the catheter position should be adjusted. As a response to this, the method may include the step of adjusting the position of the catheter in the appropriate direction to bring the middle electrode pairs closer to the diaphragm. The method steps may be repeated at regular or irregular time intervals. This will enable continuous monitoring of the catheter&#39;s position. Preferably the control unit is arranged to repeat the method steps related to stimulating, recording the signals and determining the position of the electrodes repeatedly. The stimulation of the diaphragm may be performed by invasive or non-invasive methods. Non-invasive methods have the advantage of being easier to perform and causing less discomfort to the patient. A non-invasive method would be to stimulate the phrenic nerve from the outside of the neck, but this entails a risk of stimulating another nerve instead of, or in addition to, the phrenic nerve, which may cause undesired effects. Electrical stimulation may be applied non-invasively using electrodes applied to the skin surface. Percutaneous electrodes have also been developed, which can be left in place over a period of time to allow specific reproducible stimulation patterns. The best site for transcutaneous stimulation of the phrenic nerve is on the neck a few cm above the clavicle, since this site is as far away from the vagus nerve as possible. Unwanted stimulation of the vagus nerve can cause severe bradycardia and other negative effects. Invasive methods include invasive stimulation of the phrenic nerve as well as direct invasive stimulation of the diaphragm itself. Such methods have greater impact on the patient&#39;s body but offer better control of what nerve or muscle to stimulate. Several types of implanted electrodes can be found. Invasive electrodes can be placed using several different techniques, including neck incisions, thoracoscopic procedures or thorocatomy. For example, electrodes available from Avery Labs incorporate an antenna placed immediately under the skin, which allows activation of the electrode through intact skin. European Patent application 1 389 442 discloses a neural probe having a number of electrodes, which may be used to stimulate nerve cells. Another manufacturer of suitable stimulation electrodes for diaphragm pacing is Synapse Biomedical. Atrotech Oy manufactures a phrenic nerve stimulator. The method according to the invention may be combined with other known methods for positioning the esophageal catheter. For example, the method of co-pending application No. PCT/EP2007/054149 might be used first to position the catheter. Then the inventive method may be used to adjust the catheter position. The position may be adjusted so that the electrodes located in the middle of the catheter have the highest registered EMG signals. The NEX method may also be used for a first positioning of the catheter. The inventive procedure may also be used during operation, to ensure that the catheter position has not changed too much, for example because of the patient&#39;s movements. The programming instructions of the storage medium may also cause the control unit to present the determined position of the catheter to the user on a display, for example, the display of the ventilator. This will assist the operator in determining the position of the catheter correctly. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 illustrates a patient with an esophageal catheter used to control a ventilator. FIG. 2 is a flow chart of a method according to an aspect of the invention. DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a schematic overview of a patient 1 connected to a ventilator 3 and having an esophageal catheter 5 inserted in order to record a myoelectric signal from the diaphragm. Instead of the ventilator, the inventive idea could be used with a device arranged to monitor the signal from the esophageal catheter. This myoelectric signal is fed to a control input of the ventilator 3 to control the ventilation function of the patient 1. The catheter 5 has a number of electrodes, for example, nine electrodes placed equidistantly in an array along the catheter to produce 8 subsignals, each subsignal being a difference signal between two neighbouring electrodes. The subsignals will be received by receiving means 7 and processed in a control unit 9 in the ventilator to produce the overall signal that can be used to control the ventilator. To this end, the control unit 9 is loaded with at least one non-transitory computer-readable storage medium encoded with programming instructions that control the ventilator to perform the calculations and other relevant functions. The registration of a myoelectric (EMG) signal from the diaphragm may not always be successful. As for any bioelectric signal, the EMG signal recorded from the diaphragm will comprise disturbance, in particular from the heart, but also from other muscles such as abdominal muscles. If the catheter is inserted much too far into the patient, the disturbing signals may constitute the largest part of the signal picked up by some or all the electrode pairs. In this case, there is a risk that the control signal provided to the ventilator is not related to the patient&#39;s breathing activity. In other cases, the patient may exhibit no breathing activity, or too little breathing activity to enable a proper registration. The breathing activity can be reduced, for example, because of illness or sedation. Even if the catheter is initially positioned in the right place it may be moved up or down within the patient&#39;s esophagus because of the patient&#39;s breathing activity or other movements, so that after a while the diaphragm activity is not registered in the right way. According to the invention, therefore, the arrangement of FIG. 1 also includes at least one electrode 11 for stimulating the diaphragm. The electrode 11 may be of any kind suitable for stimulating nerve or muscle cells, depending on where the stimulus is provided. The stimulus may be provided transcutaneously or subcutaneously to the phrenic nerve, or directly to the diaphragm. The electrode 11 may be controlled by an electrode control unit 13, which may be positioned in the ventilator 3, as shown in FIG. 1. It may also be a separate unit. If it is part of the ventilator, it may be integrated with the control unit 9 or it may be a separate unit. The electrode control unit 13 typically comprises a computer program for controlling the stimulation function. FIG. 1 b shows a schematic example of an esophageal catheter 5 like the one shown in FIG. 1 a. The catheter has nine electrodes, numbered e 1, e 2,..., e 9 in the Figure. Each channel is recorded as the difference signal between two adjacent electrodes, that is, between e 1 and e 2, between e 2 and e 3, etc. Hence, the uppermost channel will be the one recorded between the two uppermost electrodes e 8 and e 9, also referred to as the uppermost electrode pair. Ideally, the catheter 5 should be positioned in such a way that the electrodes e 4, e 5, e 6 located in the middle of the catheter 5 should be near the diaphragm, in order to pick up the best signal from the diaphragm. It should be understood that this configuration of the catheter is only an example. It is, however, usually suitable to place the electrodes in the middle of the catheter near the diaphragm. A method according to an aspect of the invention comprises the steps shown in FIG. 2 : Step S 20 : Position the catheter in the patient&#39;s esophagus. As discussed above, a number of methods exist for finding an appropriate position for the catheter. Step S 21 : Stimulate the muscular activity of the diaphragm at a specific point or period in time by applying a stimulus signal either to a nerve controlling the function of the diaphragm, or to the diaphragm itself. The stimulus signal may be applied as a short pulse at a point in time or over a period of time, typically during one breath. Step S 22 : Register the signals recorded by the electrode pairs of the esophageal catheter at the specific point or period in time. Step S 23 : Determine which electrode pairs record the stimulus signal with the highest amplitude. Step S 24 : Are the electrode pairs that record the strongest stimulus signal located in or close to the middle of the catheter? If yes, end of procedure; if no, go to step S 25. Step S 25 : Indicate that the catheter is not in an optimal position. To do this, the control unit 9 will indicate the position of the catheter on a display. The control unit may also issue an explicit message that the catheter position should be adjusted, for example, on the display, or through an audio alarm. Step S 26 : Adjust the position of the catheter in the appropriate direction to bring the middle electrode pairs closer to the diaphragm. This will normally be done manually by health care personnel. Return to step S 21. If the answer in step S 24 is yes, this indicates that the catheter is in an appropriate position for registering the EMG signal from the diaphragm. By repeating the procedure at regular or irregular time intervals, the position of the catheter can be monitored over time. It may be advantageous to wait for a certain period of time and then repeat the procedure to ensure that the catheter remains in the right place and to adjust it if it moves. The algorithm may also be performed in other situations, for example, when the EMG signal ceases or deteriorates dramatically, or if it is detected that ventilation correctly with the patient&#39;s breathing efforts. The latter situation is discussed in co-pending Application No. PCT/SE2007/051048. The stimulation performed in step S 21 can be carried out to provide a well defined EMG signal from the diaphragm when the spontaneous activity of the diaphragm is too weak to be recorded properly. It may also be used if there is normal activity of the diaphragm, to provide a well-defined peak of the EMG signal at a specific point in time, to ensure that the signal picked up by the electrodes is really the signal from the diaphragm and not a disturbing signal from some other muscle. The timing of the stimulation performed in step S 21 should be determined based on a number of factors. From a technical point of view a period with no disturbances from other signals, such as EMG and ECG, might be preferable, to enable an unambiguous detection of the stimulus signal. If the patient has any spontaneous breathing activity, from a clinical point of view it would be suitable to synchronize the stimulus signal with the patient&#39;s inspiration phase. The point in time at which the stimulus signal is applied should be known in order to detect when the stimulus signal will be detected. In step S 24, if the initial positioning of the catheter was unsuccessful it may be that the catheter is positioned in such a way that none of the electrodes are measuring on the diaphragm. In this case, the catheter should be adjusted up or down, preferably a distance corresponding to at least the length of the catheter that is covered by the electrodes. Then the procedure should return to step S 21. It may also happen that the amplitude of the stimulus signal is too low for it to be registered properly. If a contraction or a pneumatic triggering can be detected during stimulation, it may be concluded that the amplitude is sufficiently high. If it is determined that the amplitude is too low, the amplitude may be increased until the stimulus signal is detected by the electrodes or causes a response in the patient. As will be understood, the steps S 21 -S 25 will normally be performed by software running in the control unit 9 of the apparatus. Step S 26 will normally be performed by health care personnel.

Summary: In a method and an apparatus for determining the position of an esophageal catheter that is inserted into the esophagus of a patient, the catheter having a number of electrodes, muscular activity of the diaphragm is stimulated by applying a stimulus signal that produces a myoelectrical signal in the diaphragm having a well-defined peak at a specific point in time. The myoelectrical signal is detected by respective pairs of electrodes of said catheter, and the electrode pair or pairs that detected the signal having the highest amplitude at the specific point in time is identified. If this electrode pair or these electrode pairs that detected the signal with highest amplitude are not located approximately at the middle of the catheter, an indication is emitted that the position of the catheter should be adjusted.

big_patent

en

Summarize: US would not rule out military option if Iran breaches deal, says president in letter to New York Democrat Jerrold Nadler, in bid to shore up Congress support Barack Obama is promising Democratic lawmakers that the US will continue to keep economic pressure on Iran – and keep military options open – if a nuclear deal with Tehran goes ahead. Congress does not have votes to block Iran deal, says Nancy Pelosi Read more Obama, in a letter addressed to New York Democratic representative Jerrold Nadler, said that if Iran rushes to build a nuclear weapon, “all of the options available to the United States – including the military option – will remain available”. In the letter, which has been seen by the Guardian and was first published by the New York Times, the president also says the US will uphold sanctions targeting Iran’s non-nuclear activities, such as its support for Lebanon’s Hezbollah group and what Obama calls Iran’s “destabilising role in Yemen”. “We have a wide range of unilateral and multilateral responses that we can employ should Iran fail to meet its commitments,” the letter goes on, citing the “snap back” provisions of the deal, which allow the US and its European partners to restore sanctions in the event of a breach by Tehran. Obama wrote the letter, dated 19 August, from Martha’s Vineyard, where he is in the midst of a two-week vacation. While the president has made no public appearances during his vacation, he has been privately reaching out to Democratic lawmakers in a bid to boost support for the Iran deal. Iran nuclear deal: the key points Read more On Thursday, Nancy Pelosi, the Democratic leader in the House of Representatives, said she was confident House Democrats would have the votes if necessary to see the Iran deal through. With a Senate vote looking increasingly secure for the president, Pelosi’s comments suggest it is now extremely unlikely that Congress will halt the deal. Just two Democrats in the Senate – New York senator Chuck Schumer and New Jersey senator Bob Menendez – have defied Obama by publicly rejecting the deal. Congress will vote next month on a resolution of disapproval on the accord to curb Iran’s nuclear program in exchange for billions of dollars in sanctions relief. Obama has picked up crucial support from Democrats in recent days, including Democratic senator Claire McCaskill, who said on Thursday that while the agreement was not perfect, it was clear to her that “the world is united behind this agreement, with the exception of the government of Israel”. On Friday, Nadler, who represents a heavily Jewish congressional district, came out in support of the nuclear agreement. “The only decision that matters at this moment is whether to support or reject the agreement that is on the table now, not on whether we could or should have gotten a better deal,” Nadler wrote in an statement on Medium. “I bring to my analysis the full weight of my responsibilities as a member of Congress, and my perspective as an American Jew who is both a Democrat and a strong supporter of Israel,” Nadler said. “Accordingly, I will support the agreement and vote against a Resolution of Disapproval.” In his letter to Nadler, Obama emphasised US support for Israel, saying he views the country’s security as sacrosanct. He said he was committed to deepening missile defense funding and other military cooperation with Israel. The deal with Iran, Obama wrote, “is a very good deal for the United States, for the state of Israel, and for the region as a whole … No administration has done more for Israel’s security than mine”. Associated Press contributed to this report Photo Advertisement Continue reading the main story WASHINGTON — In his most comprehensive effort to assure wavering Democrats, President Obama wrote in a letter to Congress that the United States would unilaterally maintain economic pressure and deploy military options if needed to deter Iranian aggression, both during and beyond the proposed nuclear accord. The Aug. 19 letter, obtained by The New York Times, is addressed to Representative Jerrold Nadler, Democrat of New York, but is also aimed at other Democrats with concerns about the deal. For Mr. Obama, it reflects steps the administration could take outside the agreement. The president has repeatedly said that the deal reached by Iran and six world powers cannot be changed. While many of the promises have been made before by Mr. Obama, Secretary of State John Kerry and others, White House officials say the letter represents the first time that the president himself has compiled them under his name and in writing. It commits explicitly to establishing an office within the State Department to carry out the nuclear accord. In addition, Representative Adam Schiff of California, the ranking Democrat on the House Intelligence Committee, said that the letter expanded assurances that sanctions lifted under the nuclear accord could be reimposed piece by piece, not all at once, to keep Iran in compliance. Mr. Obama’s pledge to use the multinational commission policing the accord to block Iranian procurement of nuclear-related technology is new, as is the president’s explicit pledge “to enhance the already intensive joint efforts” of the United States and Israel in the region, said Mr. Schiff, a supporter of the deal. “Should Iran seek to dash toward a nuclear weapon, all of the options available to the United States — including the military option — will remain available through the life of the deal and beyond,” Mr. Obama wrote. He pledged to increase missile defense funding for Israel, accelerate co-development of missile defense systems, and boost tunnel detection and mapping technologies. He also vowed to increase cooperation with Israel and Persian Gulf allies to counter Iran’s efforts to destabilize Yemen, its support for Hezbollah in Lebanon, and its efforts to preserve the government of President Bashar al-Assad in Syria. Advertisement Continue reading the main story The letter comes as supporters of the nuclear deal close in on the number of lawmakers they need to sustain a presidential veto of Republican-led efforts to block it. The announcements of support by moderate Democrats from Republican-leaning states, such as Senators Claire McCaskill of Missouri on Thursday and Joe Donnelly of Indiana on Wednesday, have changed the dynamic of the Iran debate. “This was not an easy call. It was a close call,” Ms. McCaskill said Thursday. “It’s wrong for anybody, including the president, to accuse those who don’t support this deal of bad faith or warmongering.” With 26 Senate Democrats declaring their support for the accord, and five others leaning toward supporting it, it is becoming increasingly difficult for opponents of the deal in the Senate to find the 67 votes needed to override a veto. Advertisement Continue reading the main story Even some senior Republicans are looking beyond the September showdown to the accord’s implementation. “I’ve said from Day 1 it’s a heavy lift, and the administration has been effective with Democrats in saying it’s this deal or no deal,” said Senator Bob Corker of Tennessee, the Senate Foreign Relations Committee chairman working to kill the accord. “There’s going to have to be a regional follow-on now. This cannot be implemented in a vacuum because this will be our Middle East policy.” The president and Democratic leaders are now pressing for a more convincing vote. Representative Nancy Pelosi of California, the minority leader, is coordinating efforts to secure enough Democratic support to sustain a veto, through “Dear colleague” letters from respected members and assurances from outside experts. A two-thirds vote to override is also required in the House, and if the vote falls short in either chamber, the agreement goes through. “I need to have 146 to win,” Ms. Pelosi said in an interview. “Do I want more? Yes. I want more because I want this to be unifying.” So far, only two Senate Democrats — Chuck Schumer of New York and Robert Menendez of New Jersey — have declared their opposition to the deal. That raises the possibility that a resolution of disapproval, to be voted on around Sept. 16, could fail to get the 60 votes needed to break a filibuster, ending any threat of a veto showdown. Mr. Obama’s letter could solidify support. Of the 14 House and Senate Democrats who have come out against the deal, nine are from New York and New Jersey. Addressing Mr. Nadler could stanch that bleeding. In a statement, Mr. Nadler, who represents a heavily Jewish district in Manhattan, said he raised “troubling” questions “about our ability to permanently stop Iran from developing a nuclear bomb, and our commitment to strengthening the U.S. strategic relationship with Israel, as well as Iran’s continued destabilizing influence through support of terrorism and other actions that threaten Israel’s security and the security of our other Middle East allies.” “I am gratified that the president’s response satisfies a number of these concerns,” Mr. Nadler said in a statement. Mr. Nadler plans to endorse the deal on Friday, according to sources familiar with his decision, who asked to speak anonymously because the congressman has not yet made a formal announcement. Senator Chris Coons of Delaware, an influential Democrat on the Senate Foreign Relations Committee, said he and others had suggested to Mr. Obama and Vice President Joseph R. Biden Jr. ways that the administration could unilaterally address issues around the Iran nuclear accord without renegotiating it. Those include interdiction of conventional weapons shipped from Iran, enforcement of sanctions placed on Iran for its support of terrorism and its abuse of human rights, and deterrents for other destabilizing or destructive acts around the world. “This is not a one-day decision about one up-or-down vote,” said Mr. Coons, who is also undecided on whether to support the deal. “This is about what happens after the vote.” Advertisement Continue reading the main story Advertisement Continue reading the main story Republicans pounced this week on an Associated Press report that confirmed a deal between the International Atomic Energy Agency and Iran that would let Iran conduct portions of the inspections themselves at the disputed military site of Parchin, where the I.A.E.A. suspects some nuclear weapons-related experiments may have been conducted years ago. “This side agreement shows that true verification is a sham, and it begs the question of what else the administration is keeping from Congress,” said Representative Kevin McCarthy of California, the House majority leader. But that issue involved a longstanding effort by the I.A.E.A. to complete a report on past Iranian efforts to develop a nuclear weapon, an important part of the international effort to pressure Iran. It has little to do with verification of the nuclear accord between Iran and the United States, Britain, Germany, France, Russia and China. That verification regime is laid out in the actual nuclear accord and does not rely on Iran’s self-monitoring. The issue surfaced last month at the first Senate hearings on the Iran deal, when Republican senators mocked the verification procedures as unacceptable even by National Football League standards. The head of the I.A.E.A., Yukiya Amano, issued a statement Thursday saying the reports were “misleading.” He added: “I am disturbed by statements suggesting that the I.A.E.A. has given responsibility for nuclear inspections to Iran. Such statements misrepresent the way in which we will undertake this important verification work.” But because those agreements are confidential, he did not explain how the inspections would proceed. On Thursday, The Associated Press published a transcript of the original draft agreement. Mr. Corker said that by agreeing to the self-reporting procedures, the I.A.E.A. was laying down a dangerous precedent for any future disputes with Iran.

Summary: President Obama is hoping to calm concerns among Democrats still undecided or against the Iran nuclear deal. In an Aug. 19 letter to New York Democratic Rep. Jerrold Nadler written in the midst of Obama's vacation on Martha's Vineyard, the president maintains the deal itself cannot be changed, but he notes the US doesn't have its hands tied should Iran try to procure a nuclear weapon. "All of the options available to the United States-including the military option-will remain available through the life of the deal and beyond," he writes, per the New York Times. He also pledges "to enhance the already intensive joint efforts" of the US and Israel to fight Iran's support of Hezbollah and Syrian President Bashar al-Assad and its "destabilizing role in Yemen." He notes this is a "very good deal" for the US and for Israel, per the Guardian, adding, "No administration has done more for Israel's security than mine." Nadler previously raised concerns about "our ability to permanently stop Iran from developing a nuclear bomb," what the deal would mean for the US-Israel relationship, and "Iran's continued destabilizing influence through support of terrorism and other actions that threaten" Israel and the Middle East. With this letter, however, "I am gratified that the president's response satisfies a number of these concerns," Nadler says. As for why the letter was addressed to Nadler, the Times explains that 14 House and Senate Democrats publicly oppose the deal, and all but five are from New York and New Jersey. As the Times puts it, "Addressing Mr. Nadler could stanch that bleeding." To wit, sources tell the paper he'll come out in favor of the deal today. He would become the only Jewish member of Congress from New York to do so.

multi_news

en

Summarize: Thailand is renowned for its delicious curries and mouthwatering noodle dishes - so much so, many restaurants attempt to replicate these national cuisines, but with limited success. In a bid to clamp down on poorly made, bland meals, the Thai government has stepped in and created its Thai Delicious program. The initiative will use an app to find authentic recipes, alongside a so-called ‘e-delicious’ machine to test dishes across the country. The Thai Delicious program was setup to improve meals across Thailand. Its ‘e-delicious machine’ (illustrated) features 10 sensors that identify taste signatures for dishes, by measuring the conductivity of the samples at different voltages. Dishes are then given an authenticity score out of 100. The Thai Delicious program uses an app to store authentic recipes. Its so-called ‘e-delicious’ machine is then taken to restaurants to test samples of dishes. The machine has a total of 10 taste sensors. Each recipe has a unique signature, and, using these sensors, the machine can identify the signature of tested dishes. These signatures are created by measuring the conductivity of the food samples at different voltages. The two signatures, or electrical readings, are compared to one another, to achieve an authenticity score, out of 100. A score lower than 80 is a poor replica. The machine has a total of 10 taste sensors. Each authentic recipe has a unique signature, and, using these sensors, the machine can also identify the signature of tested dishes. These signatures are created by measuring the conductivity of the food samples at different voltages. These two signatures, or electrical readings, are compared to one another, to achieve an authenticity score, out of 100. Any dish that scores lower than 80 is deemed a poor replica. But those that score highly are given an award of authenticity. This includes a sticker that can be placed in the windows of restaurants. The technical specifications have not been revealed, but Nakah Thawichawatt, a Thai businessman involved in the project, told the New York Times the machines cost around £11,000 ($18,000) each. During tests, the New York Times used the machine to discover the authenticity of a green curry from the Foreign Correspondents' Club of Thailand. During tests, the New York Times used the machine to discover the authenticity of a green curry from the Foreign Correspondents' Club of Thailand (stock image pictured). It scored a below-par 78 out of 100. Last week, PhD student Joana Guerreiro from Aarhus University in Denmark released details of a nanosensor (pictured) that performs a similar ‘taste’ test for wine. Dubbed'mini-mouth', the sensor mimics the reaction that wine creates on a person's tongue to determine how the alcohol tastes and how astringent it is. It scored a below-par 78 out of 100. Last week, PhD student Joana Guerreiro from Aarhus University in Denmark released details of a nanosensor that performs a similar ‘taste’ test for wine. Dubbed'mini-mouth', the sensor mimics the reaction that wine creates on a person's tongue to determine how the alcohol tastes and, in particular, how astringent it is. The sensor uses salivary proteins to measure the sensation that would be felt on a human mouth. The researchers study how the proteins change in the interaction with the wine, and they can use this to describe the effect of the wine

Summary: The Thai Delicious program was setup to improve meals across the country. It involves an app for storing authentic recipes and an 'e-delicious machine' E-delicious has 10 sensors that create taste readings on an electrical scale. This creates a dish'signature' that can be compared to an original recipe. The machine then gives the dish a score out of 100 - and anything lower than 80 is considered poor quality. Restaurants that serve authentic Thai dishes receive an authenticity award.

cnn_dailymail

en

Summarize: A vigil for a 17-year-old who died in police custody in North Carolina ended in violence on Thursday. The march was intended as a peaceful memorial for Jesus Huerta, who died in the back of a police car of a gunshot wound to the head last month. Police, some dressed in riot gear, used tear gas and batons to disperse the crowd of about 150 friends, family and supporters as they marched towards Durham Police Department. Grief and anger: Protesters held the vigil for teen Jesus Huerta who died last month while in police custody. Police line: Officers in riot gear shadowed the protesters as they moved from Durham's downtown to the police department and back again. The supporters began at Durham City Hall and marched towards Durham Police Department, shadowed by police. According to the News Observer, as the crowd surged towards the Police Department parking lot police commanded them to leave. No answers: Jesus Huerta died last month of a gunshot wound to the head while his hands were cuffed behind his back. The crowd continued into the parking lot, some carrying signs that read 'Murdered by police.' A coordinator of the vigil, Rafael Estrada Maya called for quiet and restraint. 'We are praying! Respect prayer! Respect the dead,' he pleaded over a loudspeaker. The crowd departed the parking lot and began to move back towards Durham's downtown area where younger members of the crowd chanted and threw firecrackers at the ranks of armed riot police. Smoke bombs and tear gas were released and according to a witness, police beat protesters with sticks. 'They didn't really look like batons or night sticks, but they were thinner and longer and they were reaching over the banner whacking people with them,' said David Kaplan. 'They were clearly upset with the fact that people were out expressing themselves and upset at the fact that it appears they murdered a 17-year-old child.' Police Chief Jose Lopez said his officers were restrained in their treatment of the protesters. 'I could not be more proud of the restraint and professionalism demonstrated by our officers," Durham Police Chief Jose Lopez said in a statement to the media. 'There was a march. The peaceful intent did not exist. We used the best practices in law enforcement,' he said at a news conference Friday. At least six protesters were arrested. Broken up: Police used smoke bombs and tear gas to disperse the protesters, who had planned a peaceful vigil. Jose Huerta's sister Evelin Huerta says police. 'The actions of the Durham Police Department, led by Chief Lopez, last night were a tried-and-true tactic to intimidate and spread fear in our community,' she said. 'We call on Chief Lopez to resign immediately in light of his leadership that put dozens of armed police on the streets to scare residents and turned a memorial vigil into a war zone last night. We will not be intimidated by Chief Lopez and the Durham Police Department's tactics.' The Durham Police Department says that Jose Huerta died from a self-inflicted gunshot wound while he was sitting, handcuffed, in the back of a police cruiser. He had been taken into custody by Officer Sam Duncan for a second-degree trespassing violation at around 3am on November 19. Intimidation: Huerta's family say police tried to intimidate and spread fear among protesters who are demanding answers in the death of the teen. Force: Police pin a protester to the ground during the memorial for Jesus Huerta. According to CNN, Police Chief Lopez told a press conference that gunshot residue tests conducted on Huerta revealed a'saturation' of gunshot residue on the gloves Huerta was allegedly wearing, while Officer Duncan's hands had none. Police say the gun used in the shooting was not a police department weapon, but it is standard procedure to search suspects before they are placed in a vehicle for transport. The case is still under investigation by the State Bureau of Investigation. It was the second such event for Jesus Huerta that has erupted in violence. A protest last month ended with arrests and damage to police vehicles. The Huerta family is calling for a federal investigation and demanding that the police department release all information pertinent to the case. The family intends to hold vigils on the 19th of every month in Huerta's memory until that time

Summary: A vigil was held Thursday for a teen who died in police custody in Durham. Jesus Huerta, 17, died of a gunshot wound to the head while handcuffed in the back of a police car last month. He had been arrested on second degree trespassing charges. Police say he shot himself but family and friends are demanding an federal investigation. The supporters marched for Huerta in Durham's downtown area then towards the police department. Police used tear gas, smoke bombs and batons to disperse the protesters. At least six people were arrested. Protesters say police used unnecessary force; the police chief says officers showed restraint.

cnn_dailymail

en

Summarize: BACKGROUND OF THE INVENTION This invention relates generally to a garment bag of the type used to transport garments stored on separate hangers and more particularly to a garment bag where the hooks of the garment-bearing hangers extend through the upper end of the bag to provide a hand-grip when the bag is being transported, and to provide the point of suspension when the bag is stored, for example, on a closet rack. In the art, a small opening is generally provided centrally at the upper end of the bag for passage therethrough of the hanger hooks. Ideally the hooks are readily available to the person who wishes to transport the loaded bag. Unfortunately when the loaded bag has been lying flat, for example, on the rear seat of an automobile, or when a hanging loaded bag is lifted from the rack for purposes of hand-carriage, the hanger hooks, all or some, may slide through the top opening and become inaccessable to the user until the bag is opened. Additionally when the bag is lifted by the hanger hooks, and, as described above, one or more hangers have slipped through the top opening, the unhooked garments and their hangers fall to the bottom of the bag. Aside from the inconvenience of retrieving the fallen garments, the garment bag, designed primarily as a covering for clothing, now bears the unintended weight load of the garments. This stresses the bag walls and top and can cause tearing which most frequently occurs proximate the central top opening. What is needed is a garment bag which secures the separate and independent hangers and prevents the unintended slippage of the hanger hooks through the top opening. SUMMARY OF THE INVENTION Generally speaking, in accordance with the invention, a foldable garment bag especially suitable for transporting garments is provided. A double strap is attached to the front wall of the bag adjacent to the top central opening. The first strap member, in surface contact with the front wall, loops over the bag top and is releasably attached to the back wall. C-shaped hanger hooks protrude from the bag compartment and each hook has a substantially horizontal segment extending over the first strap member. The second strap member, overlaying and attached at the front to the first strap member, loops over the horizontal segments of the hanger hooks and is releasably secured to the first strap member. Accordingly while the strap members are secured, the hanger hooks are restrained between the strap members and are prevented from slipping into the bag compartment or from moving substantially away from the top center of the bag. When the strap members are released the hangers are individually detachable from the garment bag in the conventional manner. When the garment bag is folded, the first strap member is releasably attached near the bottom center of the folded bag, and in cooperation with lateral tabs maintains the folded condition of the bag while simultaneously restraining the hanger hook. Accordingly, it is an object of this invention to provide an improved foldable garment bag having straps capable of preventing the hanger hooks from being laterally displaced or sliding into the bag interior. Another object of this invention is to provide an improved foldable garment bag which is easily removed from a storage rack. A further object of this invention is to provide an improved foldable garment bag which supports the folded bag centrally and laterally. Still another object of this invention is to provide separable hangers with horizontal elements for cooperation with bag straps which secure the hanger to the garment bag. Still other objects and advantages of the invention will in part be obvious and will in part be apparent from the specification. The invention accordingly comprises the features of construction, combination of elements, and arrangement of parts which will be exemplified in the construction hereinafter set forth, and the scope of the invention will be indicated in the claims. BRIEF DESCRIPTION OF THE DRAWINGS For a fuller understanding of the invention, reference is had to the following description taken in connection with the accompanying drawings, in which: FIG. 1 is a front view of the garment bag with hangers of this invention; FIG. 2 is a rear view of the garment bag with hangers of FIG. 1; FIG. 3 is a view taken along the line 3--3 of FIG. 2; FIG. 4 is a partial perspective view of the garment bag of FIG. 1 in the folded condition; and FIG. 5 is a view taken along the line 5--5 of FIG. 2. DESCRIPTION OF THE PREFERRED EMBODIMENTS With reference to the Figures, the garment bag 10 of this invention includes a front wall 12, joined peripherally to the rear wall 14. A permanent joining, as by stitching (not shown) between front wall 12 and rear wall 14, continuously extends generally from the top center 16 of the bag 10, along the right shoulder 18, as seen in FIG. 1, down the right side 20, across the bottom 22, and partially up from the bottom 22 along the left side 24. A zipper fastener 26 of extended length provides a reversible closure which extends upward from the termination 28 of the permanent joining, along the left side 24 and left shoulder 30 of the bag 10 and terminates substantially at the top center 16. The zipper slide element 32 is located adjacent the top center 16 when the zipper fastener 26 is in its closed condition. Accordingly when the zipper fastener 26 is fully opened, substantially an entire side 24 and one shoulder 30 are opened for the insertion or removal of garments on hangers in the known manner. The hangers are completely separable from the bag 10. The garment bag 10 of this invention further includes a pair of tabs 34, symmetrically attached at the shoulders 18, 30 adjacent to the side joints 20, 24 and extending downward along the back wall 14. The male element 38 of a snap fastener is affixed to each tab 34, and mating female elements 40 of the snap fastener are alignedly attached to the front wall 12 near the bottom 22 of the garment bag 10. When the garment bag 10 is folded transversely (FIG. 4), the snap fastener elements 38 on the tabs 34, are releasably engaged with the fastener elements 40 on the front wall 12 to retain the bag 10 in the folded condition. The tabs 34 are fabricated from a flexible, durable material, e.g. leather, suited to carry the load imposed by the weight of the folded bag and its contents. Additional mating female elements 41 for the snap fastener are affixed to the back wall 14 in registration with the male elements 38 on the tabs 34 and are used to releasably grip the free ends of the tabs 34 when the bag 10 is not in a folded condition. A central vertical strip 42 of material, e.g. cloth, leather, is attached, for example, by stitching, to the front wall 12 to provide reinforcement for, and to enhance the appearance of the bag 10. A double strap assembly 44 is centrally attached to the front wall 12 proximate the top 16, for example, by stitching extended through the strap assembly 44, vertical strip 42 and front wall 12. The strap assembly 44 comprises a first flat, elongated, flexible, strap member 46 which is attached at one end 48 (FIG. 3) to the bag front wall 12, e.g. by stitching, as stated above. The other end 49 of the first strap member 46 is free for limited rotation about the fastened end 48 and includes a male snap fastener element 39 which when the bag is transversely folded releasably engages an aligned female snap fastener element 43 centrally attached to the front wall 14. Thus the folded-up portion of the bag 10 is supported by snap fasteners 38,40,39,43 at three locations, namely at the two lateral tabs 34 and at the central first strap member 46. When the bag 10 is extended to full length, the first strap member 46 is looped over the top of the bag 10 and connected to a female snap fastener element 45 located (FIG. 3) in registration on the back wall 14 of the bag. The second strap member 50 of the double strap assembly 44 overlays the first strap member 46 and is affixed to both the first strap member 46 and the front wall 12 proximate the fixed end 48 of the first strap member 46. The other end 52 of the second strap member is free for limited rotation about the fastened end and includes a male snap fastener element 53 which releasably engages a female fastener element 54 on the outer surface of the first strap member 46. When both strap members 46, 50 are secured by the associated snap fasteners 39, 45, 53, 54, the strap members rest with substantial surface contact between them. However, the flexible quality of the overlayed strap members 46, 50 permits the hooks of hangers to be retained therebetween without permanent distortion of the straps 46, 50 as explained hereinafter. The garment bag 10 is used in combination with separate clothing hangers 60 of generally conventional design, including downwardly sloping shoulder bars 62 connected at their center 63 and joined at the far ends 64 by a horizontal cross-bar 66. A hook 68, fabricated of stiff wire, is attached at the center 63 of the shoulder bar 62, and comprises a lower horizontal rod portion 70 (FIGS. 1, 2) extending away from the hanger center 63, and an integral rod portion 72 arching upward, and down, and passing over the center 63. Accordingly the hook 68 is substantially C-shaped and has a flat-bottomed profile as best seen in FIGS. 1 and 2. In filling the garment bag 10 with hanging garments (not shown) the zipper fastener 26 is opened and the snap fastener elements on the double strap assembly 44 are opened. The clothing hanger 60 is removed from the interior of the bag 10 and the garments are suspended. Then the hanger 60 and associated garment are passed through the zippered opening with the C-shaped hanger hook 68 extending laterally above the bag shoulders 18, 30. The horizontal rod portion 70 of the hanger hook 68 passes over the center 16 of the bag 10. The first strap member 46 is looped under the horizontal rod 70 on the hanger hook 68 and attached to the rear wall 14 by means of the snap fastener 39, 45. Then the second strap member 50 is passed over the horizontal rod 70 on the hanger hook 68 and attached to the first strap member 46 by means of the snap fastener 53, 54. Thereby the hanger hook 68 is constrained vertically between the two strap members 46, 50 and laterally in one direction by the double strap assembly 44 and in the other lateral direction by the closed shoulder joint between front wall 12 and rear wall 14. The zipper fastener 26 is then closed by bringing the sliding element 32 up adjacent to the hook 68 at the top center 16 of the bag 10. To place the loaded garment bag in a transversely-folded condition, the double strap assembly 44 is separated from the back wall 14 by opening the snap fastener 39, 45 engaging the first strap member 46. The hanger hook 68 remains secured between the two strap members 46, 50. The free ends of the tabs 34 are also separated from the back wall 14 by release from the female snap elements 41. The bag is then folded so that the snap fastener elements on the tabs 34 and first strap member 46 are in registration with the snap fastener elements 40,43 on the front wall 12. Pressure on the snap fasteners, in the known manner, releasably joins the folded-up bag portion to the upper bag portion. The loaded bag is carried, or suspended from a storage rack, by the C-shaped hooks 68. It should be noted that in using the double strap assembly 44, the stresses caused by the weight of folded garments are transmitted to the hanger hook 68 and thence via the strap members 46, 50 to the back wall 14 of the bag and to the reinforcing strip 42. Thus stresses on the zipper 26 and bag shoulders 18, 30 which might in some circumstances damage the bag 10 are relieved and directed to an area which is conveniently reinforced. It should be obvious that the steps in using the garment bag 10 of this invention are not restricted to the order presented above. For example, the zipper may be closed before the strap members are fastened. The end result is unchanged by the sequence of steps used to constrain the hanger hook 68. In alternative embodiments of this invention, the zippered opening is not limited to one bag side and a shoulder, but may extend substantially around the entire bag periphery. In another embodiment of this invention, the zippered opening may be located entirely in one wall, front or rear of the garment bag, and extending vertically, diagonally or in any direction and position suited to the insertion therethrough of hangers bearing garments It should further be understood that whereas zipper and snap fasteners have been described above in association with the garment bag 10 of this invention, other closures and fasteners, e.g. buttons, hooks, may be utilized in alternative embodiments. It will thus be seen that the objects set forth above, among those made apparent from the preceding description, are efficiently attained and, since certain changes may be made in the above construction without departing from the spirit and scope of the invention, it is intended that all matter contained in the above description or shown in the accompanying drawings shall be interpreted as illustrative and not in a limiting sense. It is also to be understood that the following claims are intended to cover all of the generic and specific features of the invention herein described, and all statements of the scope of the invention which, as a matter of language, might be said to fall therebetween.

Summary: A foldable garment bag has hooks of separable hangers extended externally from the upper end, and includes a double strap and tabs attached to the bag. One strap member passes beneath the hanger hooks and holds the folded-up portion of the bag; the other strap member loops over a horizontal, linear portion of each hanger hook and constrains the hangers to the bag. The lateral tabs cooperate with the double strap to maintain the folded condition of the bag.

big_patent

en

Summarize: BACKGROUND OF THE INVENTION 1. Field of the Invention Provided is a collection of cytofectins or cationic lipids that bind and transport polynucleotides, polypeptides, pharmaceutical substances and other biologically active species through membrane barriers. More specifically, cationic lipids are disclosed that complex with selected molecular species and facilitate delivery of those selected species into and through membranes and comparable boundary structures. 2. Description of the Background Art Cellular transfection strategies for gene therapy and similar goals have been designed and performed, but many of these procedures involve recombinant virus vectors and various problems exist with these viral gene transfer systems. Even generally advantageous adenovirus techniques encounter difficulties since most humans have antibodies to many of the adenovirus serogroups, including those that have been chosen as vectors. Wild type adenoviral superinfection of an adenoviral vector treated patient may result in propagating the recombinant vector as a defective viral particle, with the ability to infect many unintended individuals (if chosen to have a rare serogroup). The chance of adenoviral contamination is quite low but not impossible. The safety of using these genetic materials in humans remains unclear and thus hazardous. Safe, non-viral vector methods for transfection or gene therapy are essential. A few such lipid delivery systems for transporting DNA, proteins, and other chemical materials across membrane boundaries have been synthesized by research groups and business entities. Most of the synthesis schemes are relatively complex and generate lipid based delivery systems having only limited transfection abilities. A need exists in the field of gene therapy for cationic lipid species that have a high biopolymer transport efficiency. It has been known for some time that a very limited number of certain quaternary ammonium derivatized (cationic) liposomes spontaneously associate with DNA, fuse with cell membranes, and deliver the DNA into the cytoplasm and these species have been termed &#34;cytofectins&#34;. LIPOFECTIN™ represents a first generation of cationic liposome formulation development. LIPOFECTIN™ is composed of a 1:1 formulation of the quaternary ammonium containing compound DOTMA and dioleoylphosphatidylethanolamine sonicated into small unilamellar vesicles in water. Problems associated with LIPOFECTIN™ include non-metabolizable ether bonds, inhibition of protein kinase C activity, and direct cytotoxicity. In response to these problems, a number of other related compounds have been developed. The monoammonium compounds of the subject invention improve upon the capabilities of existing cationic liposomes and serve as a very efficient delivery system for biologically active chemicals. As indicated immediately above, various cationic lipids have been synthesized in previous references. For example, U.S. Pat. No. 4,812,449 discloses in situ active compound assembly of biologically active agents at target locations in preference to surroundings which are desired to be unaffected. Several charged and uncharged amine derivatives are described. Introduced in U.S. Pat. No. 5,171,678 are lipopolyamines and their use for transfecting eukaryotic cells. A polynucleotide is mixed with the subject lipopolyamine and contacted with the cells to be treated. U.S. Pat. Nos. 5,186,923 and 5,277,897 relate an enhancement of cellular accumulation of lipophilic cationic organometallic compounds by reduction of the intramembrane potential. Technetium containing compounds are disclosed. Lipophilic cationic compounds are presented in U.S. Pat. No. 5,208,036. Asymmetrical amine compounds are synthesized and employed in a method for DNA transfection. The amines are quaternized by two hydrogens or alkyl, aryl, aralkyl, quinuclidino, piperidino, pyrrolidino, or morpholine groups, unlike the present invention. U.S. Pat. No. 5,264,618 discloses cationic lipids for intracellular delivery of biologically active molecules. Asymmetric ammonium containing cationic lipids are presented for transporting molecules into membrane enclosed systems. The amines are quaternized by two hydrogens or alkyl groups, unlike the present invention. Transfection of nucleic acids into animal cells via a neutral lipid and a cationic lipid is revealed in U.S. Pat. No. 5,279,833. Liposomes with nucleic acid transfection activity are formed from the neutral lipid and the ammonium salt containing cationic lipid. U.S. Pat. No. 5,334,761 describes other amine containing cationic lipids are reported. Cationic lipids are utilized to form aggregates for delivery of macromolecules and other compounds into cells. The amines are quaternized by two hydrogens or unbranched alkyl groups, unlike the present invention. In the PCT publication of PCT/US94/13362 a heterocyclic diamine is disclosed. A symmetrical quaternary diamine having lipid tails is related for forming liposomes. The foregoing patents and publication reflect the state of the art of which the applicants are aware and are tendered with the view toward discharging applicants&#39; acknowledged duty of candor in disclosing information which may be pertinent in the examination of this application. It is respectfully submitted, however, that none of these patents teach or render obvious, singly or when considered in combination, applicants&#39; claimed invention. SUMMARY OF THE INVENTION An object of the present invention is to disclose a category of amines that greatly facilitate the delivery of biologically active compounds through membrane structures. Another object of the present invention is to present a group of cationic amine compounds that assist in the transport of selected macromolecules and other substances into and past membrane barriers. A further object of the present invention is to relate a collection of biologically active molecule transporters having the general structure: ##STR2## wherein m=1-10; R 1, R 2, and R 3 are the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, a hydroxylated alkyl, alkenyl, or alkynyl group, an ether containing alkyl, alkenyl, or alkynyl group, or a halogenated alkyl, alkenyl, or alkynyl group; R 4 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 5 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; and X - is an anion that assist in the transport of selected macromolecules and other substances into and past membrane barriers. Disclosed are novel monoamine cationic transporter molecules (and one diamine derivative) that facilitate the delivery of such compounds as polynucleotides, polypeptides, and the like into and beyond membrane walls. Generally related are compounds having the structure: ##STR3## wherein m=1-10; R 1, R 2, and R 3 are the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, a hydroxylated alkyl, alkenyl, or alkynyl group, an ether containing alkyl, alkenyl, or alkynyl group, or a halogenated alkyl, alkenyl, or alkynyl group; R 4 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 5 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; and X - is an anion. Other objects, advantages, and novel features of the present invention will become apparent from the detailed description that follows, when considered in conjunction with the associated drawings. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows a comparison of cytofectin-mediated DNA transfection using NIH 3T3 cells. FIG. 2 presents an in vivo comparison of cytofectin-mediated DNA transfection in Balb-C mice. FIG. 3 shows transfection in vivo data for various amine cytofectins. DESCRIPTION OF THE PREFERRED EMBODIMENT Referring now to the following disclosure and to the data presented in FIGS. 1-2, there are described preferred embodiments of a cationic monoamine having at least a pair of lipoyl moieties selected from a group consisting of an alkyl chain, an alkenyl chain, and an alkyl or alkenyl containing acyl chain such as: ##STR4## wherein m=1-10; R 1, R 2, and R 3 are the same or different and are hydrogen, an alkyl group, an alkenyl group, an alkynyl group, a hydroxylated alkyl, alkenyl, or alkynyl group, an ether containing alkyl, alkenyl, or alkynyl group, a halogenated alkyl, alkenyl, or alkynyl group, or acyl or acyloxy containing alkyl, alkenyl, or alkynyl group; R 4 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 5 is an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; and X - is an anion. The extra, with m more than 1, number of methylenes is introduced by standard procedures that complement the described subject synthetic pathways. A first preferred structure is: ##STR5## wherein for Compound A: n=1-10, usually between 1 and 3, preferably 1; R 3 is a hydrogen, an alkyl group, an alkenyl group, an alkynyl group, or a hydroxylated alkyl, alkenyl, alkynyl group, often an alkyl group of from 1 to 10 carbons, preferably a methyl group; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; and X - is an anion, usually a halide, and preferably iodide. ##STR6## where: the abbreviation Tr in the synthesis scheme denotes --C(Ph) 3, n=1-10, usually between 1 and 3, preferably 1; R 3 is a hydrogen, an alkyl group, an alkenyl group, an alkynyl group, or a hydroxylated alkyl, alkenyl, alkynyl group, often an alkyl group of from 1 to 10 carbons, preferably a methyl group; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; and X - is an anion, usually a halide, and preferably iodide. It is stressed that although other procedures are contemplated to be within the realm of this disclosure, a preferred method for introducing different acyl containing R 4 and R 5 groups in this compound, and in the compounds below, is the synthesis method given in &#34;A Flexible Approach to Synthetic Lipid Ammonium Salts for Polynucleotide Transfection&#34; by Bennett et al. (Tetrahedron Letters, Vol. 36, No. 13, pp. 2207-2210) and is herein incorporated by reference. In this method an acyl migration is employed to produce the mixed ester products. In the general synthesis scheme for Compound A derivatives, the first step involves reacting a tert-butyldiphenylsilyloxy derivatized material (made via a reaction of the dihydroxyethyl starting material with ClSiPh 2 tBu) with (triphenylmethoxy)methyloxirane (synthesized according to the procedure described in Bennett, M. J., Malone, R. W., and Nantz, M. H. Tetrahedron Left. 1995, 36, 2207) in the presence of lithium perchlorate in absolute ethanol. Diethyl ether in formic acid comprised the second step. The third step is a reaction with an alkyl, alkenyl, or alkynyl halide or an alkyl, alkenyl, or alkynyl containing acyl halide. The fourth step is tetrabutylammonium fluoride and THF initiated removal of the tert-butyldiphenylsilyloxy protection groups to produce the general precursor compound. The general precursor compound is then allowed to react with a selected alkyl, alkenyl, alkynyl or hydroxylated alkyl, alkenyl, or alkynyl halide. ##STR7## wherein for Compound B: n=1-10, usually between 1 and 3, preferably 1; R 3 is a hydrogen, an alkyl group, an alkenyl group, an alkynyl group, or a hydroxylated alkyl, alkenyl, alkynyl group, often an alkyl group of from 1 to 10 carbons, preferably a methyl group; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 6 is an alkyl group, an alkenyl group, an alkynyl group, or an acyl containing group all from 1 to 10 carbons, preferably a methyl group; R 7 is an alkyl group, an alkenyl group, an alkynyl group, or an acyl containing group all from 1 to 10 carbons, preferably a methyl group; and X - is an anion, usually a halide, and preferably iodide. ##STR8## where: n=1-10, usually between 1 and 3, preferably 1; R 3 is a hydrogen, an alkyl group, an alkenyl group, an alkynyl group, or a hydroxylated alkyl, alkenyl, alkynyl group, often an alkyl group of from 1 to 10 carbons, preferably a methyl group; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 6 is an alkyl group, an alkenyl group, an alkynyl group of from 1 to 10 carbons, preferably a methyl group; R 7 is an alkyl group, an alkenyl group, an alkynyl group of from 1 to 10 carbons, preferably a methyl group; and X - is an anion, usually a halide, and preferably iodide. In the general synthesis scheme for Compound B the first step involves reacting an amine starting material with (triphenylmethoxy)methyloxirane in the presence of lithium perchlorate in absolute ethanol. Diethyl ether in formic acid comprised the second step. The third step is a reaction with an alkyl, alkenyl, or alkynyl halide or an alkyl, alkenyl, or alkynyl containing acyl halide. The general precursor compound is then allowed to react with a selected alkyl, alkenyl, alkynyl or hydroxylated alkyl, alkenyl, or alkynyl halide. ##STR9## wherein for Compound C: a, b, or d are the same or different and are from 0-10, usually between 0 and 3, preferably 0 or 1; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkenyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 8, R 9, or R 10 are the same or different with each an alkyl, alkenyl, or alkynyl group or halogenated alkyl, alkenyl, or alkynyl group as long as one is halogen containing; and X - is an anion, usually a halide, and preferably iodide. More specifically for Compound C a preferred structure is: ##STR10## wherein for Compound C: a=0-10, usually between 0 and 3, preferably 1; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 8 is a halogenated alkyl, alkenyl, or alkynyl group, preferably a trifluoromethyl group; R 11 and R 12 are the same or different with each an alkyl, alkenyl, or alkynyl group or halogenated alkyl, alkenyl, or alkynyl group; and X - is an anion, usually a halide, and preferably iodide. ##STR11## where: a=0-10, usually between 0 and 3, preferably 1; R 4 and R 5 are the same or different with each an alkyl group, an alkenyl group, an alkynyl group, or an alkyl, alkenyl, or alkynyl containing acyl group; R 8 is an alkyl, alkenyl, or alkynyl group or halogenated alkyl, alkenyl, or alkynyl group, preferably a trifluoromethyl group; R 11 and R 12 are the same or different with each an alkyl, alkenyl, or alkynyl group or halogenated alkyl, alkenyl, or alkynyl group; and X - is an anion, usually a halide, and preferably iodide. In the general synthesis scheme for Compound C-1 the first step involves reacting the preferably halogenated starting material with (triphenylmethoxy)methyloxirane in the presence of lithium perchlorate in absolute ethanol. A reaction with diethylether in formic acid comprised the second step. The third step is a reaction with an alkyl, alkenyl, or alkynyl halide or an alkyl, alkenyl, or alkynyl containing acyl halide. The general precursor compound is then allowed to react with a selected alkyl, alkenyl, alkynyl or hydroxylated alkyl, alkenyl, alkynyl, halogenated R 11 and R 12 that are the same or different with each an alkyl, alkenyl, or alkynyl group or halogenated alkyl, alkenyl, or alkynyl group halide. With even more specificity, three preferred structures will now be presented with specific synthesis schemes (detailed in the Example section below). A first specific preferred structure is: ##STR12## A second specific preferred structure is: ##STR13## A third specific preferred structure is: ##STR14## There are alternate synthesis pathways for the fluorinated derivatives, two of which are presented below, but other pathways, as with the above synthesis schemes, are considered within the realm of this disclosure. ##STR15## Compound 17, immediately above, may purchased directly from Aldrich Chemical Company and is usually ordered from this source. ##STR16## Note that Compound 19 was prepared from 2,2,2-trifluoroethylamine (Aldrich Chemical Company) according to a literature procedure by Wawzonek, S., McKillip, W., and Peterson, C. J. Organic Synthesis, Coil. Vol. V 1973, 758. General Implications for Synthetic Flexibility The subject synthesis schemes present opportunities for a widely flexible array of approaches to synthesizing related amine cationic transport molecules. Not only are monosubstituted amine transporters easily synthesized by the subject procedures, but so a disubstituted and trisubstituted derivatives with like or mixed polar domain functional groups readily produced. Either a monosubstituted or disubstituted amine starting material is utilized to generate one or two functional groups in the final compound or during the quaternization step a functional group containing residue is added (see the fluoronated example above). By way of example and not by way of limitation, a mixed product is synthesized as follows: ##STR17## wherein R 30, R 40, and R 50 are the same or different and are a hydrogen, alkyl, alkenyl, or alkynyl group, a hydroxy or ether containing alkyl, alkenyl, or alkynyl group, or a halogen containing alkyl, alkenyl, or alkynyl group, R 60 and R 70 are carbonyl containing or not containing alkyl, alkenyl, or alkynyl groups, and X - is a halide (note that the initial starting material functional group or groups may need to be protected via silation or other appropriate means). More specifically, a preferred synthesis scheme for a mixed functional product is: ##STR18## wherein R 60 and R 70 are carbonyl containing or not containing alkyl, alkenyl, or alkynyl groups and X - is a halide, preferably iodide. An example of a synthesis that produces a trisubstituted derivative is as follows: ##STR19## EXAMPLES Example 1 Synthesis of N,N- Bis(2-tert-butyldiphenylsilyloxyethyl)!amine Compound 2 in an above Specific Synthesis Scheme To a mixture of diethanolamine (4.78 g, 7.26 mmol), triethylamine (2.5 mL), and 4-dimethylaminopyridine (89 mg, 0.73 mmol) in dichloromethane (73 mL) at 0° C. was added tert-butylchlorodiphenylsilane (5.46 g, 18.14 mmol). On complete addition, the reaction mixture was allowed to warm to room temperature. After 12 h, the reaction mixture was transferred to a separatory funnel and the organic layer was washed successively with saturated aqueous sodium bicarbonate, water, and brine. The organic layer was dried (sodium sulfate), filtered, and the filtrate solvent removed in vacuo. The crude product so obtained was purified by silica gel column chromatography (1% methanol in dichloromethane) to yield 2.53 g (2.13 mmol, 29%) of 2 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 7.70-7.34 (m, 20H), 3.79 (t, J=5 Hz, 4H), 2.79 (t, J=5 Hz, 4H), 2.09, (s, 1H), 1.05 (s, 18H); 13 C NMR (75 MHz, CDCI 3 ) d 135.5, 133.6, 129.6, 127.6, 63.5, 51.7, 26.9,19.2; IR (KBr) 3071, 2930,1428 cm -1. Example 2 (±)- (TriPhenylmethoxy)methyl!oxirane, Compound 3 in an above Specific Synthesis Scheme To a mixture of (±) glycidol (4.00 g, 33.5 mmol), triethylamine (5.7 mL), and 4-dimethylaminopyridine (420 mg, 3.40 mmol) in dichloromethane (170 mL) at 0° C. was added triphenylmethyl chloride (16.5 g, 51.2 mmol). On complete addition, the reaction mixture was allowed to warm to room temperature. After 12 h, the reaction mixture was transferred to a separatory funnel and the organic layer was washed successively with saturated aqueous sodium bicarbonate, water, and brine. The organic layer was dried (sodium sulfate), filtered, and the filtrate solvent removed in vacuo. The crude product so obtained was purified by silica gel column chromatography (3% diethylether in hexane) to yield 8.70 g (27.5 mmol, 82%) of 3 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 7.47-7.20 (m, 15H), 3.33-3.30 (m, 1H ), 3.16-3.09 (m, 3H), 2.76 (m, 1H), 2.61 (dd, J=2, 5H,1 H); 13 C NMR (75 MHz, CDCI 3 ) d 143.8,128.6,127.9,127.8,127.1, 127.0, 86.7, 64.7, 51.0, 44.6; IR (KBr) 3057, 2922, 1448 cm -1. Example 3 (±)-3- N,N-bis(2-tert butyldiphenylsilyloxyethyl)amino!-1-(Triphenylmethoxy)-2-propanol, Compound 4 in an above Specific Synthesis Scheme To a mixture of (±)-(triphenylmethoxy)methyloxirane (7.66 g, 24.2 mmol) and lithium perchlorate (5.87 g, 55.2 mmol) in absolute ethanol (110 mL) was added amine 2 (11.7 g, 20.2 mmol). The reaction mixture was warmed to 65° C. and allowed to stir for 24 h. After this time, the reaction solution was allowed to cool to room temperature and then transferred to a separatory funnel containing diethylether (100 mL). The resultant mixture was sequentially washed with saturated aqueous sodium bicarbonate, water, and brine. The organic layer was dried over sodium sulfate, filtered and the filtrate was concentrated by rotary evaporation to give the crude product as a yellow oil. Purification was accomplished by SiO 2 column chromatography (3% methanol in dichloromethane) to yield 14.5 g (16.1 mmol, 80%) of 4 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 7.65-7.20 (m, 25H), 3.73-3.56 (m, 5H), 3.17 (dd, J=5, 9 Hz, 1H), 2.97 (dd, J=5, 9 Hz, 1H), 2.69 (m, 5H), 2.45 (dd, J=10, 12 Hz, 1H ), 1.02 (s, 18H); 13 C NMR (75 MHz, CDCI 3 )d 144.1, 135.5, 134.7, 133.5, 129.6, 128.7, 128.6, 127.7, 127.6, 126.8, 86.4, 67.2, 66.1, 62.1, 58.4, 56.6, 26.8, 26.5, 19.0; IR (KBr) 3445, 3069, 2930, 1427 cm -1. Example 4 (±)-3- N,N-Bis(2-tet-butyldiphenylsilyloxyethyl)amino!-1,2-propanediol, Compound 5 in an above Specific Synthesis Scheme To a mixture of amine 4 (8.43 g, 9.40 mmol) in diethylether (12 mL) was added 85% formic acid (35 mL). The resulting reaction mixture was stirred at room temperature for 20 h. After this time, solid NaHCO 3 was added to neutralize the acidic solution. The resultant mixture was subsequently diluted with diethylether (100 mL) and transferred to a separatory funnel. The organic layer was separated and sequentially washed with water, and brine. Purification was accomplished by SiO 2 column chromatography (3% methanol in dichloromethane) to yield 3.75 g (5.73 mmol, 61%) of 5 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 7.65-7.31 (m, 20H), 3.68-3.60 (m, 6H), 3.40 (dd, J=4, 9 Hz, 1H), 2.71 (m, 4H), 2.57 (d, J=7 Hz, 2H), 1.03 (s, 18H); 13 C NMR (75 MHz, CDCI 3 ) d 135.5, 133.4, 129.7, 127.7, 68.0, 64.4, 62.0, 57.2, 56.7, 26.8, 19.0; IR (KBr) 3432, 3070, 2931, 1428 cm -1. Example 5 (±)-3- N,N-Bis(2-tert-butyldiphenylsilyloxyethyl)amino!-1,2-bis(9(z)-octadecenoyloxy)propane, Compound 6 in an above Specific Synthesis Scheme To a mixture of diol 5 (4.78 9, 7.26 mmol), triethylamine (2.5 mL), and 4-dimethylaminopyridine (89 mg, 0.73 mmol) in dichloromethane (73 mL) at 0° C. was added dropwise oleoyl chloride (5.46 g, 18.14 mmol). On complete addition, the reaction mixture was allowed to stir at 0° C. for 4 h whereupon an additional portion of dichloromethane (20 mL) was added. The reaction mixture was then transferred to a separatory funnel and the organic layer was washed successively with saturated aqueous sodium bicarbonate, water, and brine. The organic layer was dried (sodium sulfate), filtered, and the filtrate solvent removed in vacuo. The crude product so obtained was purified by silica gel column chromatography (6% EtOAc in Hexane) to yield 2.53 g (2.13 mmol, 29%) of 6 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 7.67-7.34 (m, 20H), 5.37 (m, 4H), 5.03 (m, 1H), 4.29 (dd, J=3, 12 Hz, 1H), 4.06 (dd, J=6, 12 Hz, 1H), 3.65 (t, J=6, 4H), 2.67 (m, 6H), 2.23 (m, 4H), 2.02 (m, 8H), 1.51 (m, 4H), 1.29 (m, 40), 1.05 (s, 18H), 0.90 (t, J=5 Hz, 6H); 13 C NMR (75 MHz, CDCI 3 ) d 173.3, 172.9, 135.5, 133.6, 130.0, 129.8, 129.7, 129.6, 127.6, 127.5, 70.0, 63.5, 62.5, 57.0, 55.4, 34.3, 34.05, 31.9, 30.0, 29.8, 29.7, 29.5, 29.4, 29.3, 29.2, 29.1 (2), 27.4, 27.2, 27.0, 26.8, 24.9 (2), 22.7, 19.1, 14.1; IR (KBr) 3071, 2927, 1741 cm -1. Example 6 (±)-3- N,N-Bis(2-hydroxyethyl)amino!-1,2-bis(9(z)-octadecenoyloxy)propane, Compound 7 in an above Specific Synthesis Scheme To a solution of amine 6 (2.50 g, 2.10 mmol) in THF (11 mL) at 0° C. was added dropwise a solution of tetrabutylammonium fluoride (6 mL of a 1M solution in THF, 6 mmol). The reaction was stirred at 0° C. for 15 h at which time analysis by thin layer chromatography revealed that no starting material was present. The reaction mixture was diluted with dichloromethane and transferred to a separatory funnel. The reaction mixture was washed sequentially with saturated aqueous sodium bicarbonate, water, and brine. The resultant organic layer was dried over sodium sulfate, filtered and the filtrate solvent removed in vacuo. The crude product was passed through a short column of silica gel using 5% methanol in methylene chloride to yield 1.03 g (1.45 mmol, 69%) of 7 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 5.34 (m, 4H), 5.18 (m, 1H ), 4.36 (dd, J =3, 12 Hz, 1), 4.10 (dd, J=6, 12 Hz, 1H ), 3.60 (t, J=5 Hz, 4H), 2.71 (m, 6H), 2.32 (dd, J=7, 14 Hz, 4H), 2.00 (m=8H), 1.61 (m, 4H), 1.37-1.15 (m, 40H), 0.87 (t, J=6 Hz, 6H); 13 C NMR (75 MHz, CDCI 3 ) d 173.7, 173.5, 129.9, 129.7, 129.6, 70.0, 63.5, 59.8, 57.2, 55.8, 34.3, 34.0, 31.9, 29.7 (2), 29.6 (2), 29.5, 29.4, 29.3, 29.1 (2), 27.2, 27.1, 24.8, 22.6, 14.1; IR (KBr) 3416, 2925, 1740 cm -1. Example 7 (±)-N,N- Bis(2-hydroxyethyl)!-N-methyl-N- 2,3-bis(9(z)-octadecenoyloxy)propyl!ammonium chloride (DODHP), Compound 1 in an above Specific Synthesis Scheme To a sealed tube containing amine 7 (0.40 g, 0.56 mmol) was added iodomethane (3 mL). The tube was flushed with argon then sealed. The reaction mixture was heated to 80° C. for 15 h. After this time, the reaction mixture was concentrated under a stream of argon (Caution: perform evaporation in a fume hood). The resulting yellow oil was dissolved in methylene chloride and transferred to a round bottomed flask. This mixture was concentrated by rotary evaporation to insure that all residual iodomethane was removed. The crude product was passed through a short silica gel column (gradient, 5%-10% methanol in dichloromethane) to yield 0.47 g (0.55 mmol, 98%) of 1 as a wax. 1 H NMR (300 MHz, CDCI 3 ) d 5.69 (m, 1H ), 5.32 (m, 4H), 4.47 (dd, J=3, 12 Hz, 1H ), 4.25-4.12 (m, 5H), 3.95-3.76 (m, 6H), 3.36 (s, 3H), 2.57 (s, 2H), 2.37 (m, 4), 1.99 (m, 8H), 1.58 (m, 4H), 1.37-1.24 (m, 40H), 0.86 (t, J=6 Hz, 6H); 13 C NMR (75 MHz, CDCI 3 ) d 173.2,172.7,129.9,129.5 (2), 65.5 (2), 63.9, 63.3, 55.6, 51.2, 34.2, 33.9, 31.8, 29.7, 29.5, 29.4, 29.2, 29.1, 29.0 (2), 27.1, 24.7, 24.6, 22.6, 14.0); IR (KBr). Example 8 (±)-1-(TriPhenylmethoxy)-3- N,N-bis(2-methoxyethyl)amino!-2-propanol, Compound 10 in an above Specific Synthesis Scheme To a mixture of oxirane 3 (5.00 g, 15.8 mmol) and lithium perchlorate (3.36 g, 31.6 mmol) in absolute ethanol (80 mL) was added amine 9 (2.53 9, 19.0 mmol). The reaction mixture was warmed to 65° C. and allowed to stir for 24 h. After this time, the reaction solution was allowed to cool to room temperature and then transferred to a separatory funnel containing diethylether (20 mL). The resultant mixture was sequentially washed with saturated aqueous sodium bicarbonate, water, and brine. The organic layer was dried over sodium sulfate, filtered and the filtrate was concentrated by rotary evaporation to give the crude product as a yellow oil. Purification was accomplished by SiO 2 column chromatography (3% methanol in dichloromethane) to yield 6.46 g (14.18 mmol, 90% ) of 10 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 7.49-7.22 (m, 15H), 3.82 (m, 1H), 3.44 (m, 4H), 3.34 (s, 6H), 3.22 (dd, J=6, 9 Hz, 1H), 3.06 (dd, J=6, 9 Hz, 1H), 2.89-2.71 (m, 5H), 2.57 (dd, J=9, 13 Hz, 1H); 13 C NMR (75 MHz, CDCI 3 ) d 144.0,128.7, 127.7,126.9, 71.2, 67.8, 66.0, 58.7, 58.4, 54.7; IR (KBr) 3437, 3058, 2874, 1449 cm -1. Example 9 (±)-3- N,N-Bis(2-methoxyethyl)amino!-1,2-propanediol Compound 11 in an above Specific Synthesis Scheme To a mixture of amine 10 (2.86 g, 6.28 mmol) in diethylether (13.4 mL) was added 85% formic acid (16.7 mL). The resulting reaction mixture was stirred at room temperature for 20 h. After this time, NaHCO 3 was added to neutralize the acidic solution. The resultant mixture was subsequently diluted with diethylether () and transferred to a separatory funnel. The organic layer was separated and sequentially washed with water, brine, and dried (sodium sulfate). Purification was accomplished by SiO 2 column chromatography (3% methanol in dichloromethane) to yield 0.99 9 (4.64 mmol, 74%) of 11 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 3.68 (m, 2H), 3.53-3.39 (m, 6H), 3.33 (s, 6H), 2.86-2.70 (m, 5H), 2.64 (d, J=6 Hz, 2H); 13 C NMR (75 MHz, CDCI 3 ) d 71.1, 68.7, 64.7, 58.7, 57.7, 54.8; IR (KBr) 3413, 2876 cm -1. Example 10 (±)-3- N,N-Bis(2-methoxyethyl)amino!-1,2-bis(9(z)-octadecenoyloxy)propane, Compound 12 in an above Specific Synthesis Scheme To a mixture of diol 11 (0.30 g, 1.41 mmol), triethylamine (0.5 mL), and 4-dimethylaminopyridine (17.2 mg, 0.14 mmol) in dichloromethane (14 mL) at 0° C. was added dropwise oleoyl chloride (1.10 g, 3.66 mmol). On complete addition, the reaction mixture was allowed to stir at 0° C. for 4 h whereupon an additional portion of dichloromethane (10 mL) was added. The reaction mixture was then transferred to a separatory funnel and the organic layer was washed successively with saturated aqueous sodium bicarbonate, water, and brine. The organic layer was dried (sodium sulfate), filtered, and the filtrate solvent removed in vacuo. The crude product so obtained was purified by silica gel column chromatography (1% methanol in dichloromethane) to yield 150 mg (0.21 mmol, 15%) of 12 as an oil. 1 H NMR (300 MHz, CDCI 3 ) d 5.31 (m, 4H), 5.08 (m, 1H), 4.35 (dd, J =3, 12 Hz, 1), 4.09 (dd, J=6, 12 Hz, 1), 3.40 (t, J=6 Hz, 4H), 3.29 (s, 6H), 2.76-2.68 (m, 6H), 2.26 (m, 4H), 1.98 (m, 8H), 1.58 (m, 4H), 1.35-1.22 (m, 40H), 0.85 (t, J=6 Hz, 6H); 13 C NMR (75 MHz, CDCI 3 ) d 173.3, 173.0, 129.9, 129.6, 71.4, 70.1, 63.7, 58.7, 55.2. 54.8, 34.3, 34.1, 31.8, 29.7, 29.6, 29.5, 29.2, 29.1, 29.0, 27.0 (2), 24.8, 22.6,14.0; IR (KBr) 2925, 2854, 1740 cm -1. Example 11 (±)-N,N-Bis(2-methoxyethyl)-N-methyl-N- 2,3-bis(9(z)-octadecenoyloxy)propyl!ammonium chloride (DODMP), Compound 8 in an above Specific Synthesis Scheme To a sealed tube containing amine 12 (150 mg, 0.20 mmol) was added iodomethane (3 mL). The tube was flushed with argon then sealed. The reaction mixture was heated to 80° C. for 15 h. After this time, the reaction mixture was concentrated under a stream of argon (Caution: perform evaporation in a fume hood). The resulting yellow oil was dissolved in methylene chloride and transferred to a round bottomed flask. This mixture was concentrated by rotary evaporation to insure that all residual iodomethane was removed. The crude product was passed through a short silica gel column (gradient, 5%-10% methanol in dichloromethane) to yield 162 mg (0.19 mmol, 95%) of 8 as a wax. 1 H NMR (300 MHz, CDCI 3 ) d 5.59 (m, 1H), 5.24 (m, 4H), 4.40 (dd, J=3, 12 Hz, 1H), 4.13-3.75 (m, 11H), 3.34 (m, 9H), 2.25 (m, 4H), 1.91 (m, 8H), 1.51 (m, 4H), 1.27-1.15 (m, 40H), 0.78 (m, 6H); 13 C NMR (75 MHz, CDCI 3 ) d 172.8, 172.6, 129.8, 129.4, 129.4, 65.9 (2), 63.5, 63.2, 63.0, 59.2, 50.4, 34.1, 33.8, 31.7, 29.5 (2), 29.3, 29.2 (2), 29.1, 29.0, 28.9 (2), 29.8, 27.0 (2), 24.5, 24.4, 22.5, 13.9; IR (KBr) 3004, 2925, 1744 cm -1. Example 12 (±)-N-(2,2,2-Trifluoroethyl)-N,N-dimethyl-N- 2,3-bis(9(z)-octadecenoyloxy)gropyl!ammonium chloride (DOFEP), Compound 14 in an above Specific Synthesis Scheme To a sealed tube containing amine 15 (0.50 g, 0.77 mmol) in DMF (5 mL) was added 2-iodo-1,1,1-trifluoroethane (1.1 mL). The tube was flushed with argon then sealed. The reaction mixture was heated to 100° C. for 15 h. After this time, the reaction mixture was transferred round bottom flask and the volatiles (DMF, excess ICH 2 CF 3 ) were removed via distillation at reduced pressure. The resulting yellow oil was passed through a short silica gel column (gradient, 5%-10% methanol in dichloromethane) to yield 67 mg (0.07 mmol, 10%) of 14 as a solid. 1 H NMR (300 MHz, CDCI 3 ) d 5.59 (m, 1H), 5.33 (m, 4H), 4.51 (m, 2H), 4.13 (dd, J=6, 12, 1), 3.87 (dd, J=9, 14 Hz, 1), 3.53 (s, 6H), 2.35 (m, 4H), 1.99, (m, 8H), 1.59 (m, 4H), 1.29-1.25 (m, 40H), 0.87 (t, J=7 Hz, 6H);; 13 C NMR (75 MHz, CDCI 3 ) d 173.0, 172.5, 129.9, 129.8, 129.5, 129.4, 66.0, 65.6, 62.8, 54.6, 34.1, 33.8, 31.7, 39.7, 29.6, 29.4 (2), 29.3, 29.1 (2), 29.0 (2), 28.9, 27.1, 27.0, 24.6, 24.5, 22.5, 13.9); IR (KBr). Example 13 Liposome formulation. An appropriate mass of the cationic lipid and a neutral lipid (DOPE) were added as solutions in chloroform to 1.9 mL sample vials to yield a 50:50 molar ratio of cationic lipid:neutral lipid. The chloroform was removed via rotary evaporation at 37° C. The resulting thin lipid films were placed under vacuum overnight to insure that all traces of solvent have been removed. The lipid mixture was resuspended in 1 mL sterile water at 70° C. until the film is hydrated, and then vortex mixed to afford an emulsion (unsonicated preparation). These emulsions were formulated at a cationic lipid concentration of 1 mM. To form the sonicated preparations used in this study, the lipid emulsions were sonicated using a Branson sonifier 450 sonicator equipped with a cup horn and recirculating water bath (35° C., 80% output with 2 sec delays over 15 minutes.). By performing comparative transfection experiments, it was determined that sonication of cytofectin emulsions above their phase transition temperature did not significantly alter transfection efficacy. Furthermore, sonication at or above 70° C. resulted in partial lipid decomposition as determined by thin layer chromatography. Example 14 Cell culture. NIH 3T3 cells were obtained from ATCC (CRL 1658), cultured in Dulbecco&#39;s Modified Eagle&#39;s Medium with 10% calf serum, and plated on standard 24 well tissue culture plates 12 to 24 hours prior to transfection. Cells were approximately 80% confluent at the time of transfection. CHO cells (ATCC CCL 61) were cultured using Ham&#39;s F12 medium supplemented with 10% fetal calf serum, and plated as described for NIH 3T3. Example 15 Transfection of cultured cells. NIH 3T3 cells were plated onto 24 well tissue culture plates as described above. The growth media was removed via aspiration and the cells were washed once with 0.5 mL PBS/well. The liposome/DNA complexes were formed through sequential addition of appropriate amounts of DMEM (serum-free), plasmid DNA (4 micrograms), and the liposome formulation into a 2 mL Eppendorf tube to a total volume of 800 microliters. Typically, 24 microliters of a lipid emulsion (1 mM cytofectin, 1 mM DOPE) were used to complex 4 micrograms of DNA to yield a 2:1 cytofectin to DNA molar charge ratio. The addition of these substances was followed by thorough vortex mixing and incubation for 15 minutes at room temperature. A 200 microliter aliquot of the resultant transfection complex was added to each well (1 microgram DNA/well, n=4) and the is cells were incubated for 4 hrs. at 37° C. At this time, 500 microliters of the appropriate growth media+10% calf serum/well was added and the cells cultured for approximately 48 hours prior to lysis and analysis. The sample transfections were subsequently repeated a minimum of three times for each cell line in order to ensure reproducibility. Example 16 Intratracheal instillation of DNA or lipid/DNA complexes. Female Balb/C mice (specific pathogen free) weighing approximately 20 to 21 grams were obtained from Charles River Laboratories. Anesthesia was provided for invasive procedures and animals were terminated by CO 2 inhalation in accordance with University of California, Davis guidelines. DNA was prepared for instillation by dilution in sterile water. Lipid/DNA complexes were prepared by mixing 20 micrograms of plasmid DNA (Luciferase) at a 4:1 molar charge ratio (cationic lipid:DNA) in sterile water for injection (total volume of 240 microliters). Mixtures were prepared and vortex mixed at room temperature, and injected within 5 minutes of lipid:DNA complex formation. Neck dissections were performed on anesthetized mice using a 1 cm incision through the skin of the anterior neck, dissection of the salivary gland and musculature surrounding the anterior trachea immediately below the larynx, and instillation of 240 microliters of DNA or lipid/DNA complex using a 1/2&#34; 30 g needle inserted 1-3 tracheal ring interspaces inferior to the larynx. After injection, the salivary gland was placed over the tracheal defect, and the superficial neck wound closed with staples. Mice were killed 48 hours after treatment and a tracheal/lung block dissected, homogenized in lysis buffer, and assayed for luciferase protein as described below. Mock treated mouse lung/trachea was used for assessment of background luciferase activity. No activity was detected in control mock-treated mouse tissue. Example 17 Luciferase assay. Relative luciferase activity was determined by using the Enhanced Luciferase Assay Kit and a Monolight 2010 luminometer (both from Analytical Luminescence Laboratories, San Diego, Cailf.). This was accomplished by directly applying 233.3 mL of concentrated luciferase lysis buffer (final concentration 0.1M potassium phosphate pH 7.8, 1% Triton X-100, 1 mM DTT, 2 mM EDTA) to each well and placing the cells on ice for 15 minutes. Removal of growth media was not necessary prior to the application of the lysis buffer. This technique enhances reproducibility by avoiding the possibility of cell loss during media removal. An analogous experiment where the growth media was removed afforded similar results. Luciferase light emissions from 31 mL of the lysate were measured over a 10 second period, and results were expressed as a function of an assumed total lysate volume of 933.3 mL. Activity has been expressed as relative light units, which are a function of assay conditions, luciferase concentration, luminometer photomultiplier tube sensitivity and background. Under the conditions described above, relative light units are related to luciferase protein mass by the equation fg luciferase=(RLU/48.6)-824!. Example 18 Transfection Data Interpretation of FIGS. 1-3 is facilitated by reference to the following list of abbreviations, prefixes, or suffixes and the associated structures: ##STR20## FIG. 1 shows a comparison of cytofectin-mediated DNA transfection using NIH 3T3 cells. DNA transfections were performed in quadruplicate as described in the experimental procedures using a 2:1 molar charge ratio (lipid charge to DNA phosphate charge). The data demonstrates that the incorporation of a dihydroxyethyl substituted ammonium functionality in the lipid polar domain leads to significantly higher transfection efficacy in vitro. Results are summarized in bar graph form as the mean (n=4) and standard deviation of total luciferase light units (RLU) obtained from cells lysed after transfection of 1 microgram of DNA. All cytofectins were formulated at a 1:1 molar ratio with DOPE. In FIG. 2 there is shown an in vivo comparison of cytofectin-mediated DNA transfection. Balb-C mice were transfected with plasmid DNA using various cytofectins. Intratracheal instillations of cytofectin:DNA complexes were performed as described in the experimental procedures. The data demonstrates that the incorporation of dimethoxyethyl and triflouroethyl substituted ammonium functionality in the lipid polar domain leads to significantly higher transfection efficacy in vivo. Results are summarized in bar graph form as the mean (n=4) and standard deviation of total luciferase light units (RLU) obtained from trachea/lung blocks lysed 48 hours after treatment with 20 micrograms of DNA. The transfection activity of various compounds is illustrated in FIG. 3. As can be seen in FIG. 3, the dioleoyl derivatives of -FEP and DMP of the subject invention are exceptionally effective in transfection. General transfection conditions were as above. The invention has now been explained with reference to specific embodiments. Other embodiments will be suggested to those of ordinary skill in the appropriate art upon review of the present specification. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the appended claims.

Summary: For use in transporting biologically active species into and through membrane barriers, a cationic amine compound is utilized that has the general structure: ##STR1## wherein R 4 and R 5 are a pair of same or different lipoyl moieties selected from a group consisting of an alkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, or alkynoyl groups and for R 1 R 2, and R 3 at least two are hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl groups or at least one amine bonded halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group or a mixture of at least one halogen containing moiety selected from a group consisting of a halogenated alkyl, alkenyl, or alkynyl group and at least one hydroxylated, ether containing, or acyloxy containing alkyl, alkenyl, or alkynyl group, and X - is an anion.

big_patent

en

Summarize: By. Matt Chorley and Jason Groves. PUBLISHED:. 17:52 EST, 21 October 2012. |. UPDATED:. 05:29 EST, 23 October 2012. Split: Andrew Mitchell resigned last week after a month of controversy. The Prime Minister was forced to defend his handling of the Andrew Mitchell affair yesterday, insisting the former Chief Whip’s rant at Downing Street police was not serious enough to cost him his job. The Prime Minister said Mr Mitchell finally resigned because of the ‘bigger question’ of whether he would be able to do his job properly, having had his authority over MPs undermined. Mr Cameron – widely criticised for allowing the issue to drag on for a month – appeared irritated as he was questioned over his refusal to sack the minister in a series of TV interviews yesterday. Mr Mitchell has denied police claims he called officers ‘plebs’ and ‘morons’ after they asked him to use a side entrance to Number Ten, rather than opening the main gate for him and his bicycle. But he admitted swearing during the altercation and finally quit on Friday night after senior colleagues told him and the Prime Minister he could not continue. Mr Cameron, speaking on ITV’s The Agenda programme last night, said: ‘The decision I had to make right at the beginning was: Was it right for him to apologise and did the police accept that apology and therefore could he get on and do his job? I took that decision. Some people would criticise that and say, “Why not just sack him straight away?” I don’t think that would have been the right thing to do. But obviously we found after a period of time he couldn’t do his job so he had to resign.’ He added: ‘I think as PM you’ve got to... if someone has apologised you need to give them a chance to do their job. I tried, whether it was Liam Fox’s resignation, I tried... rather than just do the easy thing quick at the lobby meeting at 11 o’clock meeting and have the person out the door which Tony Blair sometimes did.’ ‘It's the easiest thing in the world as Prime Minister to just sack someone at the drop of a hat when something goes wrong. I thought the right thing to do was to make sure there was a proper apology,’ he said. ‘The police didn't want to take it further but it did become apparent he wasn't going to be able to do his job so the right conclusion was reached. ‘It takes longer to discover whether someone can or can't do their job. It's much easier just to fire people, I actually think that is not the right approach.’ Yesterday Education Secretary Michael Gove revealed Mr Cameron did not believe Mr Mitchell should have lost his job because of ‘seven seconds of madness’ in Downing Street. The revelation raised fresh questions about Mr Cameron’s judgment among Tory MPs – who are still reeling from Mr Mitchell’s dramatic resignation after four weeks of controversy about his foul-mouthed rant at police officers. Michael Dugher, Labour's shadow Cabinet Office minister, claimed the PM had not learned any lessons from 'this shambles'. He added: 'While everyone else can see how wrong Andrew Mitchell was, the Prime Minister is still completely out of touch. David Cameron should stop trying to blame other people for his own weakness and dithering - and start taking some responsibility.' Mr Gove risked reopening the row with the Police Federation, by suggesting that the officers’ recollection of former Chief Whip Mr Mitchell’s rant may have been wrong. He said: ‘There’s a Japanese film, I think it’s called Rashomon, in which different participants who see the same event all have different recollections of it. I wasn’t there. I trust Andrew, and I’m always inclined to give him the benefit of any doubt.’ One Tory MP said yesterday he was ‘staggered’ that Mr Mitchell had not been sacked as soon as news broke of his confrontation. Another said the Prime Minister had been ‘misguided’, while a third accused him of presiding over a ‘catastrophe’ that would do lasting damage to the party’s image. In a further blow, former Conservative Party chairman Lord Tebbit accused Mr Cameron of leading a ‘dog of a coalition government’ that had let itself be portrayed as ‘a government of unfeeling toffs’. Mr Mitchell finally quit the Cabinet on Friday night, saying he had lost the backing of the Parliamentary party. Scroll down for video. Opposed: David Cameron does not believe Andrew Mitchell should have lost his job because of ‘seven seconds of madness’ in Downing Street, Michael Gove (right) revealed last night. Senior ministers, including Home Secretary Theresa May, Work and Pensions Secretary Iain Duncan Smith and Mr Mitchell’s own deputy John Randall, were among those who told him he should go for the good of the party. Mrs May made no attempt to deny her role yesterday, telling the BBC: ‘I’m not going to talk about private conversations. Andrew has resigned. I think that is an end to the issue.’ Accepting Mr Mitchell’s resignation on Friday, Mr Cameron hinted at a possible comeback. And Mr Gove revealed that the Prime Minister still believed Mr Mitchell should be in the Cabinet. Home Secretary Theresa May told Mr Mitchell he should resign. He told Sky News: ‘David Cameron wanted to keep Andrew, I wanted Andrew to stay because I don’t believe and the Prime Minister doesn’t believe that 30 years of public service should be effaced at a stroke by seven seconds of unacceptable but very human exasperation. ‘But Andrew came to that decision and he did so because he wanted to put the interests of the party collectively ahead of his own.’ Mr Gove repeated the claim in a later interview, saying the Prime Minister had ‘wanted him to stay on’. He acknowledged that the row had overshadowed good news on the economy last week, but said the public would see through the ‘froth’, noting that Tony Blair had won three elections despite a string of scandals involving ministers in his government. Mr Gove’s comments came as backbenchers broke cover to voice their dismay at the Prime Minister’s handling of recent events. Tory MP Andrew Percy said he had told his whip as soon as the original story broke that Mr Mitchell ought to be removed because it would ‘run and run and run’. ‘I was frankly a bit staggered at the time that Downing Street hadn’t picked up on the whole mess earlier,’ he said. Mr Percy added: ‘A lot of the problems we are facing at the moment are not problems about the economy but actually it’s stuff we have made ourselves. The disasters or shambles that are created wholly within Westminster or within Downing Street.’ Brian Binley, secretary of the backbench 1922 Committee, described Mr Cameron’s decision to bring back Old Etonian Sir George Young to replace Mr Mitchell as a ‘comfort-zone appointment’. He added: ‘I’m afraid it points to very poor man-management – David is the sort of guy who wants people around him who is comfortable with rather than people who will challenge him.’ MP Philip Davies said Mr Cameron had been ‘misguided’ to hang on to Mr Mitchell, adding: ‘It was handled badly. Andrew Mitchell should have resigned, I think when he didn’t resign the Prime Minister should have sacked him.’ In an article for the Observer, Lord Tebbit wrote: ‘This dog of a coalition government has let itself be given a bad name and now anybody can beat it. ‘It has let itself be called a government of unfeeling toffs. Past governments have had far more real Tory toffs: prime ministers Alec Douglas-Home and Harold Macmillan, or even in Thatcher’s day, Whitelaw, Soames, Hailsham, Carrington, Gowrie, Joseph, Avon, Trenchard and plenty more, without incurring similar abuse.’

Summary: The Prime Minister says it would have been 'easy' to sack his minister but the 'proper thing to do' was get him to say sorry. Education Secretary Michael Gove suggested he and the PM had tried to persuade Mr Mitchell to 'tough it out' Mr Mitchell's dramatic resignation was due to foul-mouthed rant at police officers as he tried to exit the Downing Street gates on his bike. Mr Gove suggested the officers' recollection of rant may have been wrong.

cnn_dailymail

en

Summarize: Introduction Congress has granted many federal agencies the authority to issue regulations that carry the force of law. This grant of authority raises the issue of how those agencies should be held accountable for the regulations they implement. One method of maintaining accountability is requiring agencies to analyze the potential effects of new regulations—sometimes called regulatory analysis or regulatory impact analysis —before implementing them and making the analyses public during the rulemaking process. An important and commonly performed type of regulatory analysis is a cost-benefit analysis (CBA)—a systematic examination, estimation, and comparison of the economic costs and benefits resulting from the implementation of a new rule. By performing and making public such analyses, an agency demonstrates that it has given reasoned consideration to the necessity and efficacy of a rule and the effects it will have on society. Most agencies regulating the financial industry are not subject to certain statutes or other requirements that apply to most executive branch agencies, allowing them to operate with a relatively high degree of independence from the President and Congress. These financial regulators—along with other agencies that have similar independence—are often referred to as independent regulatory agencies. Agencies are given this independence in part so that experts writing technical rules have some degree of insulation from political considerations. One aspect of these regulatory agencies' independence is that they are not subject to certain requirements that direct other agencies to perform CBAs with certain parameters and executive review. Some observers argue that this independence is appropriate and that subjecting financial regulators to increased requirements would inhibit implementing necessary, beneficial regulation. However, others argue that financial regulators should be subject to greater requirements to increase accountability. The debate has drawn increased attention in recent years as regulators promulgate and continue to promulgate rules mandated and authorized by the Dodd-Frank Wall Street Reform and Consumer Protection Act ( P.L. 111-203 ). In response, a number of bills in recent Congresses have proposed increased requirements. This report examines issues related to financial regulators and CBAs, including potential difficulties facing such regulators and methods available to them when preforming a CBA; the analytical requirements the agencies currently face; and the arguments for and against increasing requirements on financial regulators. This report also briefly describes several examples of proposed legislation that would change the requirements facing financial regulators. Overview of Cost-Benefit Analysis CBA can help ensure that regulators demonstrate that their decisions are based on an informed estimation of likely consequences during the development, issuance, and implementation of rules. In the analysis, economists and other experts use theory, modeling, statistical analysis, and other tools to estimate the likely outcomes if a particular regulation were to be implemented. These outcomes are compared with the likely outcomes if no regulation or a different regulation were implemented. Then the good outcomes (benefits) can be weighed against bad outcomes (costs) of a regulatory action to determine whether and to what degree a regulation is on net beneficial to society. Benefits may include such outcomes as deaths and injuries avoided, acres of rare habitat saved, or a decreased probability of financial crisis. Costs may include outcomes such as increased production costs for companies, regulation compliance cost to companies, and increased prices for consumers. Externalities —the effects experienced by parties that are not directly involved in the market transactions covered by the regulation—also should be included in the analysis to the extent possible. If it were the case that regulators were expected to make decisions with complete information, all societal costs and benefits would need to be accurately and precisely estimated. These outcomes would be quantified (assigned accurate numerical values) and monetized (assigned an accurate dollar value). Proposed rules would be finalized and implemented only if benefits were expected to exceed costs, and in a form that maximized net benefits. However, societal costs and benefits may be difficult to accurately estimate, quantify, and monetize. Therefore, performing most CBAs involves some degree of subjective human judgement and uncertainty, and predicted results are often expressed as a range of values. As discussed in more detail in the " Financial Regulator Requirements Debate " section, some argue that performing CBAs for financial regulation is particularly challenging, due largely to the high degree of uncertainty over precise regulatory costs and outcomes. This raises questions about the appropriate scope, level of detail, and degree of quantification that should be required of analysis performed in the rulemaking process. On one hand, overly lenient requirements could allow agencies to implement overly burdensome regulation with limited benefit without due consideration of consequences. In addition, a CBA can be an informational tool that estimates the potential effects of a rule and informs the agency and the public as various groups advocate for certain policies—and potentially exaggerate or minimize risks, costs, or likely outcomes of a certain regulation. In contrast, overly onerous analytic requirements could risk impeding the implementation of necessary, beneficial regulation because performing the analysis would be too time consuming, too costly, or simply not possible. Another concern is that if agencies face highly burdensome requirements, they may have an incentive to achieve policy goals through other methods—such as issuing policy statements, guidance documents, and technical manuals—that create less accountability than the rulemaking process. In addition, a CBA itself can be costly and is performed by departments and agencies funded by general taxes and fees on industry. Finally, requiring uncertain and contestable CBAs may allow self-interested parties to impede socially beneficial regulation by challenging agency analysis in court and offering their own subjective analysis. For these reasons, stringent CBA requirements may themselves generate more costs than benefits. Current Cost-Benefit Analysis Requirements As mentioned above, CBA can be a useful tool for ensuring good regulations are implemented and that regulatory agencies are accountable. However, requirements to perform such analyses may restrict agencies from effectively regulating. This section examines current CBA requirements, including those that apply to nonfinancial regulators and those that direct financial regulators more specifically. It also reviews certain government reports examining the methods and results of recent regulatory CBAs performed by financial regulators under the existing requirements. Requirements for Nonfinancial Regulators: Executive Order 12866 and OMB Circular A-4 The primary requirement for most agencies to calculate estimates of costs and benefits when issuing rules is under Executive Order (E.O.) 12866, which was issued in 1993 by President William Clinton. E.O. 12866 requires covered agencies—that is, agencies other than independent regulatory agencies, which includes most of the financial regulators—to submit "significant" rules to the Office of Management and Budget's Office of Information and Regulatory Affairs (OIRA) for review, along with an initial cost and benefit assessment. For rules that are determined to be significant because their annual economic effect is likely to exceed a $100 million threshold, covered agencies are required to conduct a more in-depth CBA. Specifically, the order requires agencies to provide to OIRA an assessment of anticipated costs and benefits of the rule, and an assessment of the costs and benefits of "reasonably feasible alternatives" to the rule. Other E.O. 12866 provisions encourage agencies to consider costs and benefits during the rulemaking process for all rules, although those other provisions do not require a complete, detailed cost-benefit analysis for non-economically significant rules. E.O. 12866 has remained in effect since 1993, and it was reaffirmed in 2011 in E.O. 13563 by President Barack Obama. E.O. 13563 states that covered agencies should (1) propose or adopt a regulation only upon a reasoned determination that its benefits justify its costs, (2) tailor regulations to impose the least burden on society, and (3) select regulatory approaches that maximize net benefits. It also directs agencies to "use the best available techniques to quantify anticipated present and future benefits and costs as accurately as possible." In September 2003, OMB finalized Circular A-4 on regulatory analysis, which refined and replaced an earlier OMB guidance document, providing good-guidance practices to agencies for conducting their CBAs. The circular states that it was "designed to assist analysts in the regulatory agencies by defining good regulatory analysis... and standardizing the way benefits and costs of Federal regulatory actions are measured and reported." The document provides some specific information that agencies should generally include in their analyses, such as the statutory or judicial directives that authorize the action; the underlying problem or market failure prompting the regulation; consideration of a "reasonable number" of regulatory alternatives; and both a cost-benefit analysis and a cost-effectiveness analysis. Circular A-4 remains the current OMB guidance for agencies preparing CBAs under E.O. 12866 requirements. Exception for Independent Regulatory Agencies from Executive Order 12866 The exception for independent regulatory agencies in Executive Order 12866 is similar to the exception found in Executive Order 12291, in which President Ronald Reagan first established centralized regulatory review in OIRA and required cost-benefit analysis of certain regulations in 1981. This decision is widely understood to have been based on political considerations regarding the statutorily designed independence of these agencies. In short, President Reagan—and subsequent Presidents—viewed these agencies as having been designed by Congress to be independent of the President, and as such chose not to subject them to presidential (OIRA) review. The statutory categorization of those agencies had been codified in the Paperwork Reduction Act of 1980, which designated a special set of procedures for those agencies' information collection approvals from OMB. E.O. 12291, and later E.O. 12866, referenced the PRA's list of agencies to identify the excepted agencies. Currently, the list of independent regulatory agencies includes the following financial regulators: Board of Governors of the Federal Reserve System, Commodity Futures Trading Commission, Federal Deposit Insurance Corporation, Federal Housing Finance Agency, Securities and Exchange Commission, Bureau of Consumer Financial Protection, Office of Financial Research, Office of the Comptroller of the Currency, and National Credit Union Administration. When President Clinton issued Executive Order 12866 in 1993, he, like President Reagan, chose to exempt the independent regulatory agencies from the order's CBA requirements. Similarly, President Obama continued to exempt independent regulatory agencies from CBA requirements with E.O. 13563, although his OIRA Administrator encouraged those agencies to "give consideration to all [E.O. 13563's] provisions" in a memorandum issued soon after the executive order. In July 2011, President Obama issued E.O. 13579, "Regulation and Independent Regulatory Agencies." The executive order encouraged independent regulatory agencies to comply with some of the principles in E.O. 13563 that were directed to Cabinet departments and independent agencies (e.g., public participation, integration and innovation, flexible approaches, and science). In a separate memorandum issued the same day as the executive order, the President said he was taking these actions with "full respect for the independence of your agencies." E.O. 13579 did not, however, directly apply the cost-benefit principles in E.O. 12866 and 13563 to independent regulatory agencies, nor did it require these regulators to conduct CBA before issuing their rules. CBA Requirements on Financial Regulators As previously discussed, the financial regulators are exempt from many of the analytical requirements and guidance documents that are applicable to executive agencies, including E.O. 12866 and OMB Circular A-4. However, financial regulators may be required to conduct CBA or other regulatory analyses under cross-cutting statutes or pursuant to the underlying statutes that provide them with rulemaking authority. Requirements facing financial regulators arguably require a relatively narrow analysis or allow for more agency discretion compared to the requirements discussed above under E.O. 12866. For example, agencies may be required to "consider" or "estimate" costs, benefits, or other economic effects, but the degree to which those considerations must be quantified and monetized estimates is not specified. However, the requirements facing financial regulators are not trivial, and financial regulations have been vacated following judicial review when the court found the CBA performed during rulemaking to be deficient. Cross-Cutting Analytical Requirements The following statutes contain analytical requirements that apply to all federal regulatory agencies, including the financial regulators. Regulatory Flexibility Act The Regulatory Flexibility Act (RFA) of 1980 ( P.L. 96-354 ) requires federal agencies to assess the impact of their forthcoming regulations on "small entities," which the act defines as including small businesses, small governmental jurisdictions, and certain small not-for-profit organizations. Under the RFA, all regulatory agencies, including the financial regulators, must prepare a "regulatory flexibility analysis" at the time proposed and certain final rules are issued. The RFA requires the analysis to describe, among other things, (1) the reasons why the regulatory action is being considered; (2) the small entities to which the proposed rule will apply and, where feasible, an estimate of their number; (3) the projected reporting, recordkeeping, and other compliance requirements of the proposed rule; and (4) any significant alternatives to the rule that would accomplish the statutory objectives while minimizing the impact on small entities. However, these analytical requirements are not triggered if the head of the issuing agency certifies that the proposed rule would not have a "significant economic impact on a substantial number of small entities." Paperwork Reduction Act The Paperwork Reduction Act (PRA) of 1980 ( P.L. 96-511 ) pertains to certain aspects of the rulemaking process, albeit not the rules themselves. The PRA's primary purpose is to minimize the paperwork burden for individuals, small businesses, and others resulting from the collection of information by or for the federal government, which often stems from regulatory requirements: many information collections, recordkeeping requirements, and third-party disclosures are contained in or are authorized by regulations as monitoring or enforcement tools. In fact, these paperwork requirements are sometimes a primary component of requirements stemming from financial regulation. The PRA requires agencies to justify any collection of information from the public by establishing the need and intended use of the information, estimating the burden that the collection will impose on respondents, and showing that the collection is the least burdensome way to gather the information. Paperwork burden is most commonly measured in terms of "burden hours." The burden-hour estimate for an information collection is a function of the frequency of the information collection, the estimated number of respondents, and the amount of time that the agency estimates it takes each respondent to complete the collection. Agencies must receive OIRA approval (signified by an OMB control number displayed on the information collection) for each collection request before it is implemented, and those approvals must be renewed at least every three years. OIRA can disapprove any collection of information if it believes the collection is inconsistent with PRA requirements. However, multiheaded independent regulatory agencies can, by majority vote of the leadership, void any OIRA disapproval of a proposed information collection. Analytical Requirements Applicable Solely to Banking Regulators The Riegle Community Development and Regulatory Improvement Act (Riegle Act) imposes analytical requirements on rulemaking for the federal banking regulators—the Federal Reserve, the Office of the Comptroller of the Currency (OCC), and the Federal Deposit Insurance Corporation (FDIC). One of the Riegle Act's primary purposes is to reduce administrative requirements for insured depository institutions, and the scope of the analysis required reflects that specific aim. When determining the effective date and compliance requirements of new rules that impose additional reporting, disclosure, or other requirements on depository institutions, the federal banking regulators must take into consideration: "(1) Any administrative burden that such regulations would place on depository institutions, including small depository institutions and customers of depository institutions; and (2) the benefits of such regulations." Agency-Specific Requirements for CBA Certain individual agencies—the Securities and Exchange Commission (SEC), the Consumer Financial Protection Bureau (CFPB), and the Commodity Futures Trading Commission (CFTC)—are statutorily required to perform certain analysis in rulemaking specific to the agency. As mentioned previously, the parameters of analysis when "considering" cost and benefits are to a degree left to agency discretion, although analysis could be subject to judicial review if a party were to challenge the regulation in court. The SEC is subject to requirements to analyze the effect of its rules, with an emphasis on market efficiency and competition. The National Securities Market Improvement Act ( P.L. 104-290 ) requires the SEC to "consider or determine whether an action is necessary or appropriate in the public interest... [and] whether the action will promote efficiency, competition, and capital formation." The Securities Exchange Act (P.L. 73-291) requires the SEC to perform economic analysis on "the impact any such rule or regulation will have on competition." The CFPB must specifically consider the costs and benefits to consumers and the companies to which the new rules apply. The Dodd-Frank Wall Street Reform and Consumer Protection Act (Dodd-Frank Act) ( P.L. 111-203 ) requires the CFPB to " consider (1) the potential benefits and costs to consumers and covered persons, including the potential reduction of access by consumers to consumer financial products or services resulting from such rule; and (2) the impact of proposed rules on covered persons... and the impact on consumers in rural areas." The CFTC must evaluate costs and benefits of new rules and the analysis must include several specified considerations. The Commodity Exchange Act (P.L. 74-675) requires the CFTC to "consider the costs and benefits of the action" before promulgating a rule, and "the costs and benefits of the proposed Commission action shall be evaluated in light of: (A) considerations of protection of market participants and the public; (B) considerations of the efficiency, competitiveness, and financial integrity of futures markets; (C) considerations of price discovery; (D) considerations of sound risk management practices; and (E) other public interest considerations." Cost-Benefit Analysis in Practice Reports on the characteristics of the agency-performed CBAs—including independent regulatory agencies—can illustrate what analyses are done in practice as part of rulemaking. Section 624 of the Treasury and General Government Appropriations Act of 2001 (31 U.S.C. §1105 note)—sometimes known as the "Regulatory Right-to-Know Act"—requires OMB to issue an annual report to Congress on regulatory costs and benefits. The report generally includes an assessment of the CBAs for major rules done by agencies as a part of rulemaking. The 2016 report indicated that independent financial regulatory agencies issued 8 major final rules during FY2015, and that although benefits and costs were considered during the rulemaking process for all these rules, they were not always monetized. Six of these rules provided monetized costs, but none provided monetized benefits. In comparison, executive departments and agencies subject to E.O. 12866 implemented 30 major rules: 21 analyses monetized both benefits and costs; 6 monetized costs but not benefits; 2 monetized benefits but not costs; and 1 did not monetize costs or benefits. The Government Accountability Office (GAO) releases an annual report on Dodd-Frank regulations that examines analyses done by financial regulators. These reports typically make an assessment of the degree to which the analyses—for rulemaking related to Dodd-Frank provisions—were consistent with the directives of OMB Circular A-4, even though the regulators are not required to follow the directives. In general, GAO has found that financial regulator analysis is consistent with that guidance. For example, in the 2016 report, GAO notes, Independent federal financial regulators are not required to follow OMB's Circular A-4 when developing regulations, but they told us that they try to follow this guidance in principle or spirit. Regulators generally included the key elements of OMB' s guidance in their regulatory analyses for these major rules. To assess the extent to which the regulators follow Circular A-4, we examined 5 major rules... Specifically, we examined whether the regulators (1) identified the problem to be addressed by the regulation; (2) established the baseline for analysis; (3) considered alternatives reflecting the range of statutory discretion; and (4) assessed the costs and benefits of the regulation. We found that all five rules we reviewed were consistent with OMB Circular A-4. Challenges and Variants of Cost-Benefit Analysis CBA of any type of regulation faces challenges in making an accurate assessment of the regulation's effects. Over recent decades, academics and agency experts have developed sophisticated and useful techniques to do these types of analyses, but they generally contain a degree of uncertainty. Some challenges include behavioral changes of people as they adapt to a new regulation, which are difficult to predict; quantification that must overcome uncertainty over the causal relationship between the regulation and outcomes; and monetization, which is difficult for outcomes that do not have easily discernable monetary values. Variations of CBAs address some of these difficulties, including cost-effectiveness analysis, which compares costs of alternative regulation when benefits cannot be accurately quantified or monetized; breakeven analysis, which can establish the likelihood or under what conditions a regulation would be beneficial; qualitative analysis with expert judgement, in which experienced professionals describe and explain likely effects that cannot be quantified and make a judgement as to how costs compare with benefits; and retrospective analysis, which estimates the realized costs and benefits following some period of time—often years— after implementation of rules. This section examines these challenges and variants as they relate to CBA generally. There is debate over whether the challenges are particularly daunting for financial regulation CBA and to what degree different types of analysis can solve these problems. An examination of the arguments related to financial regulator CBA requirements can be found in the following section, entitled " Financial Regulator Requirements Debate." Challenges of CBA One difficulty in performing cost-benefit analysis is trying to accurately determine the human behavioral response to the implementation of a regulation. For example, consider a hypothetical and very simplified CBA that analyzes a new requirement that financial institutions make additional disclosures to customers about a certain type of loan. To estimate the benefit to consumers who avoided entering into a bad financial arrangement, the analysis would have to estimate, among other things, how many potential customers would read the disclosure and would elect not to use the product on the basis of that information. Of these, how many would then seek out a substitute credit source? Predicting human choices such as these involves modeling consumer behavior in this market, statistical interpretations of available data, and some degree of uncertainty. Quantification of outcomes also poses challenges in determining causation and measuring magnitudes of effects. Returning to the hypothetical regulation outlined above, suppose lenders also would be required to report additional performance data, such as default rates, about the loans. The additional cost of reporting could decrease loan profitability. In such a case, lenders will likely reduce the availability of these loans. An important cost of this regulation might be reduced economic growth by the contraction of credit. Making an estimation of this cost would involve macroeconomic modeling, statistical interpretation, and uncertainty. After an estimate has been made of the quantity and magnitude of outcomes, those effects must be monetized because measuring the varied effects of a regulation requires a common unit of measurement. This becomes problematic when attempting to assign a dollar value to outcomes that do not have market prices. For example, imagine a proposed regulation aimed at reducing the number of home foreclosures. An important benefit might be the avoidance of the emotional distress families may experience as a result of being forced to move from their homes and finding alternative housings. Assigning a dollar value to this outcome would require sophisticated techniques and would likewise involve uncertainty. Finally, regulatory benefits may often be more difficult to monetize than major costs. Costs are often economic costs, which may be more easily monetized, such as an industry's reduction in economic activity or the added expense of complying with regulation. Benefits may be harder to quantify because of the difficulty in determining causation and because the outcomes are harder to price. Financial regulation benefits that may be difficult to monetize include the emotional distress of foreclosure cited in the previous example, consumer and investor confidence in knowing they are protected from fraud, and decreased probability of a financial crisis. Variants of CBA Quantified and monetized estimates generally provide the clearest measurement and comparison of the costs and benefits of proposed regulation. However, variants of CBA can be performed when full quantification and monetization is not entirely possible due to the challenges described above. Some of these variants include cost-effectiveness analysis, breakeven analysis, and qualitative analysis with expert judgement. Also, agencies sometimes do retrospective analysis. Although not a part of rulemaking and so beyond the scope of this report, it deserves mention because this type of analysis is the subject of proposals to assess the regulatory system and identify regulations that should be amended or repealed. Cost-Effectiveness Analysis Cost-effectiveness analysis may be useful if benefits of a regulation are hard to monetize. In these analyses, an outcome is identified as necessary or sufficiently important to the advancement of social welfare, such as preventing cancer cases, preserving wetlands, or reducing the likelihood of financial crises. A set of alternative regulations—ranging from stringent to lenient—is then analyzed to determine how well each alternative achieves the objective outcome and at what cost. This comparison is useful for identifying the most effective form of regulation. Breakeven Analysis Breakeven analysis may be useful when estimates of either benefits or costs or both face a relatively large degree of uncertainty, and the estimates fall within a wide range. In these analyses, the magnitudes of the quantified costs and benefits are compared to determine what values of the unquantified variables would have to be for the regulation to break even or impose no net cost on society. The analysis—in the face of a relatively high level of uncertainty—can reveal under what circumstances a regulation would benefit society or at least identify which regulations are most or least likely to do so. For example, consider another highly stylized analysis of a hypothetical regulation aimed at reducing cases of a certain disease. The cost of the regulation is estimated to be $50 million; how many cases would be avoided can only be estimated in the range of 10,000-50,000; and monetizing the benefit of avoiding a case is problematic. Given these hypothetical values, the breakeven value of avoiding one case of the disease is between $1,000 and $5,000. To use extreme examples for the purpose of illustration: if this disease is the common cold, it could be argued that the regulation is overly burdensome; but if the disease is fatal, it could be argued that the regulation should be implemented. Qualitative Analysis with Expert Judgement Wherever benefits and costs cannot be quantified to a reasonably informative degree of certainty and precision, they could be analyzed qualitatively. This analysis type describes the factors considered, the rationale used in making a policy choice, and the regulators' professional judgement in assessing the regulation's welfare effects. Retrospective Analysis Retrospective analysis estimates the realized costs and benefits following some period of time—often years— after implementation of rules. This analysis eliminates some uncertainties about what outcomes will be observed under the regulation. However, the results of the analysis still involve assumptions and uncertainty in assessing the degree to which the regulation caused the observed outcomes or estimating what outcomes would have been realized if the regulation had never been implemented. Retrospective analysis is different from most of the analysis covered in this report, in that it is an ex post analysis performed after implementation and so cannot be part of the rulemaking process. Financial Regulator Requirements Debate Most observers agree that performing CBA is often a useful tool for the regulatory rule-writing process. However, whether financial regulators should be required to perform CBAs with specified parameters that would be subject to review is a matter of long-standing debate, probably at least in part due to their exemption from E.O. 12866. In addition, the issue may have attracted increased attention in recent years as many financial regulations have been implemented in response to the financial crisis, particularly after the enactment of the Dodd-Frank Wall Street Reform and Consumer Protection Act. Some observers argue that financial regulators should maintain a relatively high degree of discretion over the role and form of CBAs in the rule-writing process. They assert certain characteristics of the finance industry—discussed in detail below—necessitate CBAs with more easily contestable assumptions and uncertain results than in other industries; and performing highly contestable and uncertain CBAs does not discipline agencies, but instead may provide an opportunity for interested parties to impede socially beneficial regulation. Others argue that financial regulators should be subject to more stringent requirements than is currently the case. They assert performing CBAs for regulation of the finance industry does not pose greater difficulties than for regulation of other industries, and imposing requirements on financial regulators would spur them to overcome methodological and other challenges; and financial CBAs—despite contestable and uncertain results—would be the best tool for ensuring that regulation is implemented responsibly with due consideration of consequences. This section presents the two sides of this debate. Arguments That Financial Regulator Discretion is Appropriate Some observers assert that performing CBAs for financial regulation is different from other types of regulation. They claim financial regulation CBAs are more uncertain and contestable, and this limits the effectiveness of CBA requirements. Therefore, the argument goes, the CBA requirements facing most regulators would not be appropriate for financial regulators. Others advocate more generally for a relatively high degree of agency discretion to use expert judgement. One potential reason for greater uncertainty in financial CBA is that the outcomes are almost wholly dependent on human behavioral responses. Unlike regulation of other sectors, the objects of regulation are not chemicals or pieces of machinery, but the activities of individuals and financial firms and their interactions in interrelated markets for intangible financial goods. The behaviors of a pollutant in an ecosystem, a drug in the human body, or material in a car during a crash are governed by biological, chemical, and physical laws. The implementation of a regulation does not change these reactions. However, the behavior and reactions in the financial system are governed by human behavior within a system of laws and regulations. A new regulation changes the system itself and its effects result entirely from human behavioral changes. This may make the effects—especially the first-order, direct effects—harder to accurately predict than in other industries. For example, if certain factories were required to install a piece of equipment that prevented the release of a pollutant, the cost of the equipment is identifiable and the direct effect of how much of the pollutant would be captured can likely be measured. In contrast, if a requirement is implemented on banks to hold more liquid assets, the cost to banks is uncertain because it depends on what types of assets banks choose to shed from their balance sheets, what they add, and what effect those actions have on the market prices of those assets. It is also unclear how to quantifiably measure the liquidity of the financial system or its resultant benefits. Another reason cited as a potential cause for uncertain estimates is the central role the financial system plays in the entire economy. For most industries, changes in factors such as production cost, price, and quantity demanded and supplied resulting from regulations can be calculated using relatively well-vetted economic models. However, the causal channels through which financial changes affect overall economic activity are complex with no consensus macroeconomic model that can be used to make precise estimates. In addition, innovation in finance—unlike innovation in industries using physical equipment and chemical processes—faces few physical constraints, possibly allowing the financial system to change more quickly than other industries. Therefore, estimating how a regulation implemented today will affect markets years in the future is challenging. For example, in the years leading up to the financial crisis, private label sub-prime mortgage securitizations and collateralized debt obligations grew very rapidly and to a level of importance in the financial system that would have been difficult to have foreseen when many regulations were being developed. Another confounding factor in financial CBA is that for many financial regulation objectives there is not always consensus about whether outcomes are benefits or costs. For example, most agree improved health outcomes are beneficial and increased consumer prices and industry cost should be counted as costs. However, the cost-benefit tally for financial regulation is sometimes not as clear cut. If a consumer protection provision is expected to reduce a certain kind of high-interest-rate lending, experts might reasonably argue over to what degree this is a benefit versus a cost; it is a benefit to the extent it reduces an abusive practice, but a cost to the extent it reduces the availability of a needed credit source. Often such a lack of clarity arises because the effects of financial regulation often consist largely of wealth transfers between various groups—such as transfers between lenders and borrowers or between businesses seeking to raise capital and investors. CBA is a tool most often used to measure the net economic effects, and economic transfers between groups are typically a secondary concern. Proponents of greater agency discretion argue that placing more stringent requirements on financial regulators for conducting CBAs could potentially make issuing regulations more costly and time consuming. Those proponents argue that increasing CBA requirements could lead agencies to block or delay the issuance of individual regulations, and that over time, this could ultimately result in less stringent regulation. Proponents of agency discretion further assert that CBAs involving such a high degree of uncertainty and contestable assumptions would not discipline agencies. Instead of increasing accountability and regulatory efficiency, they argue CBAs could disguise agency judgement as objective, scientific measurement. Instead of providing an authoritative rationale for a regulation, they argue requirements would provide an opportunity for parties aiming to protect their own interests—not social welfare—to challenge certain beneficial regulations by offering competing but similarly subjective CBAs. Arguments That Stricter Requirements on Financial Regulators Are Needed In contrast, some observers believe that regulatory analysis requirements for financial regulators are not stringent enough. Proponents of increased CBA argue that the challenges facing financial regulators are not substantively more difficult than those facing other regulators when performing CBA. They note that all regulation elicits uncertain human behavioral responses. For example, the direct effects of antitrust regulation—where CBA plays an important role—similarly are almost entirely based on the reaction of firms, consumers, and markets. They also challenge the claim that financial innovation is especially rapid compared with other industries, citing the rapid advances in agriculture and pharmaceuticals. In addition, the largest financial regulation effects may actually be easier to monetize because they largely involve changes in monetary transactions rather than health or environmental outcomes that involve assigning a dollar value to nonmarket outcomes. Proponents of stricter requirements also take issue with the argument that the centrality of finance to the economy represents a reason for exemption from CBA requirements. First, they again disagree that estimating financial effects is uniquely and prohibitively complex, noting the sophistication of CBA performed by other regulators. Next, they argue that the potential to cause very large effects across the entire economy increases the importance of CBA in financial regulation, because implementing harmful financial regulation is more consequential than if the industry were more peripheral to the economy and had small economic effects. Proponents further assert that financial regulation CBA seems to face such difficult challenges because it has been exempt from certain requirements and oversight. Other regulators—once faced with similar problems—have overcome challenges because requirements spurred them to develop agency expertise and methods for performing CBA. They argue that if faced with similar requirements financial regulators, experts, and consultants would similarly devise solutions. Some academics have already started to propose methods to address questions specific to the financial industry. Furthermore, CBA's proponents argue uncertainty and imprecision are not valid reasons for foregoing financial CBA. They note that most CBAs involve some degree of uncertainty and assumptions. Nevertheless, by requiring agencies to perform the analysis, the assumptions used in evaluating the regulation are articulated and transparent, and their merits can be evaluated. Even if estimated outcomes fall over a wide range of values, an analysis can still make an assessment of the likelihood a regulation will be beneficial and how its costs can be minimized. In these ways, they argue uncertain CBAs can play an important role in showing when a proposed regulation is hard to justify or easy to defend. Proponents argue CBAs—despite possible limitations—are the best alternative for identifying good and bad regulations and have rightly become an important and often required part of rulemaking. For these reasons, they assert financial regulators should face requirements similar to those facing regulators of other industries. Selected CBA Legislation A number of bills have been introduced and seen action in recent Congresses that would impose additional regulatory impact analysis requirements on financial regulators, including bills that would impose more stringent CBA requirements. Examples include bills that would impose certain requirements on all agencies, including the financial regulators; bills that address independent or financial regulators specifically; and bills affecting only one financial regulator. 115th Congress The Financial CHOICE Act ( H.R. 10 ) was introduced in the House on April 26, 2017. Section 312 of the bill would require financial regulators to perform certain analyses as part of the rulemaking process, including a quantitative and qualitative assessment of all anticipated direct and indirect costs and benefits of the regulation. Proposed rules found to have quantified costs greater than quantified benefits would require a congressional waiver before being implemented. The Regulatory Accountability Act ( H.R. 5 ) passed the House on January 11, 2017, and introduced in the Senate ( S. 951 ) on April 26, 2017. The bill would make several changes to the rulemaking process of all agencies by amending the Administrative Procedure Act. Among the changes, agencies would have to consider alternatives to the new regulation and the potential costs and benefits of the alternatives. The bill would extend requirements for CBA to all agencies, including independent regulatory agencies. The OIRA Insight, Reform, and Accountability Act ( H.R. 1009 ) passed the House on March 1, 2017. The bill, among other measures, would codify into law OIRA authority of reviewing agency CBA in rulemaking. This authority would also be extended to independent regulatory agencies. The SEC Regulatory Accountability Act ( H.R. 78 ) passed the House on January 12, 2017. The bill would impose additional cost-benefit requirements for the SEC, would specify parameters and considerations that must be part of the analysis, and would require the SEC to retrospectively assess the impact of adopted regulation. The CFTC Commodity End-User Relief Act ( H.R. 238 ) passed the House on January 12, 2017. The bill would expand the number of considerations that CFTC is statutorily required to include in its CBAs from 5 to 12. The additional considerations include the cost of compliance with the regulation and alternatives to direct regulation. 114th Congress The Independent Agency Regulatory Analysis Act of 2015 ( S. 1607 ) would have authorized the President to subject independent regulatory agencies to CBA requirements that exist in executive order—such as EO 12866. Notably, this would include the E.O. 12866 requirement that major rules be submitted for OIRA review with an initial cost and benefit assessment. Bills that would have imposed new analysis requirements on individual financial regulators include the Federal Reserve Accountability and Transparency Act of 2015 ( H.R. 113 ), the Fed Oversight Reform and Modernization Act ( H.R. 3189 ), and the CFPB Dual Mandate and Economic Analysis Act ( H.R. 5211 ). Conclusion Congress likely will continue to face questions over what appropriate CBA requirements for financial regulators should be. A reasoned and systematic examination of likely consequences of a regulation is a useful practice to ensure good and avoid bad regulation. However, calibrating requirements to reach this outcome is difficult. Excessively lenient requirements could allow bad regulation to be implemented, because regulators could promulgate regulations without due consideration of their likely effects. In contrast, excessively stringent requirements could block good regulation from being implemented, because the time and resources required to perform the analysis could make the cost to regulators prohibitively high. The calibration is complicated by the difficulties and uncertainties involved in performing CBA. Additional lack of clarity is involved in financial regulation, because experts disagree over whether CBA is especially difficult and uncertain in that field. These factors suggest that the question of what CBA requirements financial regulators should face may not be easily settled.

Summary: Cost-benefit analysis (CBA) in the federal rulemaking process is the systematic examination, estimation, and comparison of the potential economic costs and benefits resulting from the promulgation of a new rule. Agencies with rulemaking authority implement regulations that carry the force of law. While this system allows technical rules to be designed by experts that are to some degree insulated from political considerations, it also results in rules being implemented by executive branch staff that arguably are not directly accountable to the electorate. One method for Congress to increase accountability is to require the regulators to conduct analyses of likely effects of proposed regulations. In this way, an agency demonstrates that it gave reasoned consideration to the effects of the proposed rules. CBA is an important type of such analysis, as comparing costs and benefits can be useful in determining whether or not a regulation is beneficial. However, performing CBA can be a difficult and time-consuming process, and it produces uncertain results because it involves making assumptions about future outcomes. Some observers argue that financial regulation CBA is particularly challenging. This raises questions about what parameters and level of detail agencies should be required to include in their CBA. While most federal regulatory agencies are directed by Executive Order 12866 and Office of Management and Budget Circular A-4 in their performance of CBAs, financial regulators are generally not subject to these directives. Financial regulators are statutorily required to perform certain CBA: requirements such as the Paperwork Reduction Act (P.L. 104-13) and Regulatory Flexibility Act (P.L. 96-354) generally apply to all financial regulators; financial regulators that regulate the banking system are subject to requirements set out in the Riegle Community Development and Regulatory Improvement Act (P.L. 103-325); and agencies such as the Securities and Exchange Commission (SEC), the Consumer Financial Protection Bureau (CFPB,) and the Commodities Futures Trading Commission (CFTC) face requirements specific to them. However, the requirements facing individual financial regulators generally allow them to perform analysis under less specific instruction than is contained in the requirements that are cited above and apply to nonindependent regulatory agencies. Whether the requirements facing financial regulators should allow for this discretion is a contentious issue. Some observers assert that financial regulators should maintain a relatively high degree of discretion over when and how to conduct CBA. They argue that characteristics of the financial industry and regulation make CBAs in this area especially uncertain and contestable, and assert that financial regulation effects depend entirely on human and market reactions; finance plays a central role in a huge, complex economic system; and financial regulations' effects are more likely (relative to other types of regulation) to include transfers between groups not well accounted for in net measurements. They further argue that requisite CBAs that are uncertain and contestable are more likely to disguise agency discretion as objective fact and provide the opportunity for interested parties to challenge socially beneficial regulation with their own subjective, self-interested analyses. Other observers assert that financial regulators should face more stringent requirements than they currently do. They refute claims that financial CBAs are necessarily more uncertain or contestable than in other areas. Also, they argue that tools and techniques would be developed to overcome challenges if CBAs were required. They further argue that even uncertain and contestable CBAs are effective in disciplining agencies because they create transparency of the agency's evaluations of proposed regulations and allow for outside assessment of that evaluation. Recent Congresses have been active on this issue, and the House has passed several bills in the 115th Congress that would increase CBA requirements. Recent proposals would affect either all regulators including financial regulators, financial regulators as a group, or individual financial regulators.

gov_report

en

Summarize: President Barack Obama stepped into an oil-slicked Keystone Pipeline mess on his way out of town Friday, contradicting the findings of his own State and Energy Departments. The pipeline, a 1,179-mile long planned conduit stretching from Albert, Canada to the Gulf of Mexico, has generated environmental controversy that the administration pledged to weigh against the project's economic impact. Obama claimed Friday during a press conference before he left for an extended vacation that America would hardly notice the $8 billion enterprise's impact on their pocketbooks. 'At issue in Keystone is not American oil. It is Canadian oil that is drawn out of tar sands in Canada,' he told a packed press briefing room. Obama made no mention of oil from Montana and North Dakota that will be incorporated into the three-foot-wide pipeline's load if the project gets a green-light from Congress in the new year. SCROLL DOWN FOR VIDEO. NOT IMPRESSED: Obama said the Keystone pipeline will bring little benefit to Americans and won't include oil from American wells. The three-foot-wide pipeline, pieces of which are seen here, will wend its way through 1,179 miles of North America on a path from Alberta, Canada, to the Gulf coast of Texas. North Dakota Senator John Hoeven (left) disputes Obama's contentions about the pipeline, and has helped recruit key Democrats including West Virginia Sen. Joe Manchin to is side with arguments about energy independence. 'It would save Canadian oil companies and the Canadian oil industry an enormous amount of money if they could simply pipe it all the way through the United States down to the Gulf,' he said, casting the enterprise as a sop to businesses north of the border. Incoming Senate Majority Leader Mitch McConnell said this week that a Keystone approval bill will be his chamber's first agenda item in January. But the president hedged his bets on what he would do if a united, Republican-led Congress dropped that legislation on his desk. 'I'll see what they do,' he said. 'We'll take that up in the New Year.' A reporter asked Obama if he had been short-selling the economic benefits of the pipeline, and he shot back that 'I don’t think I’ve minimized the benefits, I think I’ve described the benefits.' And those, he said, were slim. 'There is very little impact, nominal impact, on U.S. gas prices – what the average American consumer cares about – by having this pipeline come through,' he claimed. 'There’s a global oil market. It's very good for Canadian oil companies and it's good for the Canadian oil industry, but it’s not going to be a huge benefit to U.S. consumers.' 'It’s not even going to be a nominal benefit to U.S. consumers,' Obama said, putting an exclamation point on his argument. Republican North Dakota Sen. John Hoeven fired back, telling DailyMail.com that the president's judgment was 'just not accurate.' 'The president said that the pipeline would move only Canadian oil and that it would all be exported. He’s wrong on both counts,' said Hoeven. 'The Keystone XL pipeline will also carry light sweet crude from North Dakota and Montana, and the president's own Department of Energy produced a report that said the oil would be used in the United States.' That report, a June 2011 memorandum, came from the desk of Carmine DiFiglio, the department's deputy assistant secretary for policy analysis. NORTH DAKOTA CRUDE: Rancher Bob Banderet stands in front of the Keystone pumping station that will move oil from the state into the main pipeline – something Obama said Friday won't happen. Hoeven also had an answer for Obama's insistence on Friday that the project's chief negative remains the threat of an outsize environmental footprint that could drive new levels of climate change. 'I want to make sure that if, in fact, this project goes forward, that it's not adding to the problem,' Obama said. 'If we've got more flooding, more wildfires, more drought, there are direct economic impacts on that.' But Hoeven said Obama forgot to count the environmental impact of the Alberta tar sands oil's current mode of transportation. TransCanada, the company that has been angling to build the pipeline since 2008, is shipping the oil 'through rail or trucks,' the president said during his press conference. The North Dakota senator suggested that he should read the latest environmental impact report from his own State Department. That report, he said, concluded that 'it would take 1,400 tanker rail cars every day to ship the same amount of oil as the pipeline.' 'Ironically, not building the pipeline will result in more emissions from trucks, trains and oil tankers than would ever be produced by the Keystone XL pipeline.' NOT QUITE: Sen. Mary Landrieu, a Louisiana Democrat, was one vote shy of passing a Keystone pipeline approval bill in November, and her failure helped a Republican snatch her seat away in an election runoff. It's unclear whether Obama is laying the groundwork for a veto. But ultimately the Nebraska Supreme Court could have as much to say about the outcome as the White House. Governor Dave Heineman has approved the pipeline's route through his state, but a group of landowners challenged that decision. If they prevail, TransCanada will have to start over with Nebraska's Public Services Commission and find a new path through the Cornhusker State. That could take as long as a year. Louisiana Democratic Sen. Mary Landrieu tried to make approving Keystone the cornerstone of her failed bid to keep her seat in November and December. She was among the president's allies who have deserted him on the issue. Oil refineries dot Landrieu's state like chicken pox and provide Louisianans with a recession-proof economic baseline. She tried in mid-November to marshal the 60 votes in the Senate required to break a logjam and approve the pipeline, but she fell short by just one. The House of Representatives passed its version of the bill by a wide margin

Summary: Senator John Hoeven, a North Dakota Republican, says his state and Montana are both planning to move crude oil on the $8 billion pipeline. Obama said his chief concern with the TransCanada company's project is the environmental impact. Hoeven said the Canadian oil's current transport - trucks and trains - have a bigger climate change impact than the pipeline. The president spoke at a year-end press conference before jetting to Hawaii for an extended vacation. Incoming Senate Majority Leader Mitch McConnell said this week that approving the 1,179-mile pipeline will be the Senate's first job in January. But the Nebraska Supreme Court could decide next month to reject its proposed path through the state, setting the project back months or longer.

cnn_dailymail

en

Summarize: The head of Puerto Rico’s power authority has resigned amid a scandal over efforts to restore electricity to the hurricane-hit island, its governor announced Friday. Governor Ricardo Rossello told a news conference that Ricardo Ramos had resigned as executive director of the Puerto Rico Electric Power Authority to avoid "distractions." Rossello defended Ramos’ management but suggested that his resignation was necessary in the midst of the controversy. He did not say whether or not he had asked for his resignation. More than half of the population of Puerto Rico is still without electricity nearly two months after Hurricane Maria ravaged the US territory of more than three million people. Critics of Ramos have also questioned the decision to award a tiny American firm a $300 million contract to help restore power, a deal that has since been cancelled. Story highlights Whitefish Energy's contract is up on December 1 The company says it is stopping work before then because it is owed millions of dollars San Juan, Puerto Rico (CNN) Whitefish Energy is stopping its work to restore Puerto Rico's broken electricity grid because the company says it is owed more than $83 million by the island's power authority. Whitefish CEO Andy Techmanski told CNN that repeated requests for agreed payments were not met and there was no choice but to suspend work. He claimed credit for the restoration of transmission lines by his contractors, even after his company's controversial contract with the Puerto Rico Electric Power Authority (PREPA) was set to be voided. "We stopped because of the financial situation, lack of payment with PREPA has gotten beyond its maximum threshold and what we can sustain as a business," he said. A letter sent by Whitefish to PREPA and seen by CNN accuses PREPA of delaying payments. As of Sunday, Whitefish said $83,036,305.09 was outstanding, including more than $26 million that it said had been audited and approved by PREPA already. Without payment to Whitefish, contractors and subcontractors were also going unpaid, the letter said. FILE - In this Oct. 19, 2017 file photo, a brigade from the Electric Power Authority repairs distribution lines damaged by Hurricane Maria in the Cantera community of San Juan, Puerto Rico. Puerto Rico's... (Associated Press) FILE - In this Oct. 19, 2017 file photo, a brigade from the Electric Power Authority repairs distribution lines damaged by Hurricane Maria in the Cantera community of San Juan, Puerto Rico. Puerto Rico's... (Associated Press) SAN JUAN, Puerto Rico (AP) — Whitefish Energy Holdings said late Monday that it was halting work to help restore power in Puerto Rico because the U.S. territory's government has not paid crews as part of a contract that led to accusations of overcharging and incompetence and contributed to the resignation of the power company director. The Montana-based company said in a statement that invoices for work done in October are outstanding and that it can no longer keep working. The Associated Press obtained a letter dated Nov. 19 and signed by Whitefish CEO Andy Techmanski saying that Puerto Rico's government owes Whitefish more than $83 million and that the company would suspend work on Monday if it wasn't paid. Whitefish said in the letter that the lack of payments is a breach of the $300 million contract that the administration of Gov. Ricardo Rossello cancelled last month. Even though the contract had been cancelled, both sides agreed Whitefish would complete its current projects and remain in Puerto Rico until Nov. 30. "There is no basis for PREPA to withhold payments from Whitefish Energy," the letter said, referring to the Puerto Rico Electric Power Authority. "We have met the terms of the contract — including completing difficult work on time and under challenging conditions." The company said Techmanski was not immediately available for comment. Whitefish also said Monday that Florida-based utility companies working in Puerto Rico were pulling out ahead of time. "While we cannot speak for these utilities, we have been assured by their representatives that this is about their go-forward concerns once Whitefish Energy completes its work with PREPA and the ability of PREPA or any successor organizations to provide their crews with the necessary resources and management," Whitefish said. The company said it would resume work if it obtained the payments owed. Meanwhile, Puerto Rico's government said in a statement late Monday that Whitefish is alleging nonpayment and that the power utility is reviewing and auditing Whitefish invoices. It also said the Electric Power Authority was forced to stop pending payments "until the situation with the Whitefish subcontractor is clarified." Puerto Rico power company spokesman Carlos Monroig told the AP that both sides are in talks to reach an agreement to satisfy everyone involved. The dispute comes just days after the resignation of Ricardo Ramos, the power company director who signed the Whitefish contract, which is undergoing a local and federal audit. Documents released by the U.S. House Natural Resources Committee ahead of a recent hearing show the electric utility ignored advice from its own lawyers before signing the contract with Whitefish, which is based in Interior Secretary Ryan Zinke's hometown and had just two employees when Hurricane Maria hit. Many people in Puerto Rico remain without power two months after the Category 4 storm devastated the island, killing at least 55 people and causing damage estimated at up to $95 billion. More than 20 of Puerto Rico's 78 municipalities remain in the dark, and overall power generation stands below 50 percent. Major blackouts also have hit the capital of San Juan and surrounding areas in recent weeks, for reasons ranging from overgrown vegetation to fuel not being supplied in time because of logistical limitations. The hurricane hit as Puerto Rico entered its 11th year in recession and struggles to restructure a portion of its $73 billion public debt load. The power company holds about $9 billion of that debt. Whitefish Energy company announced that it has decided to stop its work in Puerto Rico after Puerto Rico Energy Power Authority stopped payments, owing the company $83 million. Interested in Puerto Rico? Add Puerto Rico as an interest to stay up to date on the latest Puerto Rico news, video, and analysis from ABC News. Add Interest In a statement, the Montana-based company said despite the company’s “diligence and that of its subcontractors” payments under the contract with the island’s bankrupt energy authority have been delayed. Whitefish said that it will not continue any work and will not perform any additional work until PREPA pays for work that has already been completed. PREPA confirmed that it had stopped payments to Whitefish Energy after receiving “a communication from one of Whitefish's subcontractors requesting the stoppage of any payment to the company, since it owed the subcontractor money.” PREPA added that it is in communication with Whitefish and the subcontractor to resolve the situation. PREPA did not confirm the amount owed to Whitefish. PREPA signed a controversial contract with Whitefish Energy, which only has two full-time employees, for $300 million, which was canceled after public backlash. Even though the contract was canceled, the terms of the contract stated Whitefish would work for an additional 30 days and complete its projects. The date of the contract’s completion is Nov. 30. Two months after Hurricane Maria slammed into Puerto Rico, only 46.6 percent of the island’s electrical grid has power. Officials on the ground have not given a number of how many homes or businesses are receiving that power.

Summary: The Montana energy company that set up a controversial, since-canceled contract to help restore Puerto Rico's power after Hurricane Maria had agreed to work through Nov. 30 before heading out. But now it's stopping work altogether, claiming the Puerto Rico Electric Power Authority owes it upward of $83 million, ABC News and the AP report. "We stopped because... lack of payment with (Prepa) has gotten beyond its maximum threshold and what we can sustain as a business," Whitefish CEO Andy Techmanski tells CNN. But Prepa, whose director resigned Friday over the contract scandal, says it's not paying until it can resolve an issue between Whitefish and an unnamed Whitefish subcontractor. Prepa claims it received "a communication" from the subcontractor "requesting the stoppage of any payment" to Whitefish over non-payment to the subcontractor. Only about half of the island's power grid is back online two months after the hurricane, though CNN notes that doesn't mean half of the island's customers are with power. The AP reports that San Juan and nearby areas keep getting hit with blackouts. "It may have not been the best business decision coming to work for a bankrupt island," Techmanski tells CNN, adding that he thought FEMA would help ensure his firm would get paid. He says he hopes the payment can be worked out so that Whitefish can wrap up its work there through the end of November. A Prepa spokesman tells the AP negotiations are now taking place to make sure that happens. Meanwhile, the original Whitefish contract is undergoing both local and federal audits to find out how it came to pass in the first place.

multi_news

en

Summarize: The bright morning sun dazzled the eyes, the snow had ceased, the mists had vanished, the mountain air was so clear and light that the new sensation of breathing it was like the having entered on a new existence. To help the delusion, the solid ground itself seemed gone, and the mountain, a shining waste of immense white heaps and masses, to be a region of cloud floating between the blue sky above and the earth far below. Some dark specks in the snow, like knots upon a little thread, beginning at the convent door and winding away down the descent in broken lengths which were not yet pieced together, showed where the Brethren were at work in several places clearing the track. Already the snow had begun to be foot-thawed again about the door. Mules were busily brought out, tied to the rings in the wall, and laden; strings of bells were buckled on, burdens were adjusted, the voices of drivers and riders sounded musically. Some of the earliest had even already resumed their journey; and, both on the level summit by the dark water near the convent, and on the downward way of yesterday's ascent, little moving figures of men and mules, reduced to miniatures by the immensity around, went with a clear tinkling of bells and a pleasant harmony of tongues. In the supper-room of last night, a new fire, piled upon the feathery ashes of the old one, shone upon a homely breakfast of loaves, butter, and milk. It also shone on the courier of the Dorrit family, making tea for his party from a supply he had brought up with him, together with several other small stores which were chiefly laid in for the use of the strong body of inconvenience. Mr Gowan and Blandois of Paris had already breakfasted, and were walking up and down by the lake, smoking their cigars. 'Gowan, eh?' muttered Tip, otherwise Edward Dorrit, Esquire, turning over the leaves of the book, when the courier had left them to breakfast. 'Then Gowan is the name of a puppy, that's all I have got to say! If it was worth my while, I'd pull his nose. But it isn't worth my while--fortunately for him. How's his wife, Amy? I suppose you know. You generally know things of that sort.' 'She is better, Edward. But they are not going to-day.' 'Oh! They are not going to-day! Fortunately for that fellow too,' said Tip, 'or he and I might have come into collision.' 'It is thought better here that she should lie quiet to-day, and not be fatigued and shaken by the ride down until to-morrow.' 'With all my heart. But you talk as if you had been nursing her. You haven't been relapsing into (Mrs General is not here) into old habits, have you, Amy?' He asked her the question with a sly glance of observation at Miss Fanny, and at his father too. 'I have only been in to ask her if I could do anything for her, Tip,' said Little Dorrit. 'You needn't call me Tip, Amy child,' returned that young gentleman with a frown; 'because that's an old habit, and one you may as well lay aside.' 'I didn't mean to say so, Edward dear. I forgot. It was so natural once, that it seemed at the moment the right word.' 'Oh yes!' Miss Fanny struck in. 'Natural, and right word, and once, and all the rest of it! Nonsense, you little thing! I know perfectly well why you have been taking such an interest in this Mrs Gowan. You can't blind _me_.' 'I will not try to, Fanny. Don't be angry.' 'Oh! angry!' returned that young lady with a flounce. 'I have no patience' (which indeed was the truth). 'Pray, Fanny,' said Mr Dorrit, raising his eyebrows, 'what do you mean? Explain yourself.' 'Oh! Never mind, Pa,' replied Miss Fanny, 'it's no great matter. Amy will understand me. She knew, or knew of, this Mrs Gowan before yesterday, and she may as well admit that she did.' 'My child,' said Mr Dorrit, turning to his younger daughter, 'has your sister--any--ha--authority for this curious statement?' 'However meek we are,' Miss Fanny struck in before she could answer, 'we don't go creeping into people's rooms on the tops of cold mountains, and sitting perishing in the frost with people, unless we know something about them beforehand. It's not very hard to divine whose friend Mrs Gowan is.' 'Whose friend?' inquired her father. 'Pa, I am sorry to say,' returned Miss Fanny, who had by this time succeeded in goading herself into a state of much ill-usage and grievance, which she was often at great pains to do: 'that I believe her to be a friend of that very objectionable and unpleasant person, who, with a total absence of all delicacy, which our experience might have led us to expect from him, insulted us and outraged our feelings in so public and wilful a manner on an occasion to which it is understood among us that we will not more pointedly allude.' 'Amy, my child,' said Mr Dorrit, tempering a bland severity with a dignified affection, 'is this the case?' Little Dorrit mildly answered, yes it was. 'Yes it is!' cried Miss Fanny. 'Of course! I said so! And now, Pa, I do declare once for all'--this young lady was in the habit of declaring the same thing once for all every day of her life, and even several times in a day--'that this is shameful! I do declare once for all that it ought to be put a stop to. Is it not enough that we have gone through what is only known to ourselves, but are we to have it thrown in our faces, perseveringly and systematically, by the very person who should spare our feelings most? Are we to be exposed to this unnatural conduct every moment of our lives? Are we never to be permitted to forget? I say again, it is absolutely infamous!' 'Well, Amy,' observed her brother, shaking his head, 'you know I stand by you whenever I can, and on most occasions. But I must say, that, upon my soul, I do consider it rather an unaccountable mode of showing your sisterly affection, that you should back up a man who treated me in the most ungentlemanly way in which one man can treat another. And who,' he added convincingly,'must be a low-minded thief, you know, or he never could have conducted himself as he did.' 'And see,' said Miss Fanny,'see what is involved in this! Can we ever hope to be respected by our servants? Never. Here are our two women, and Pa's valet, and a footman, and a courier, and all sorts of dependents, and yet in the midst of these, we are to have one of ourselves rushing about with tumblers of cold water, like a menial! Why, a policeman,' said Miss Fanny, 'if a beggar had a fit in the street, could but go plunging about with tumblers, as this very Amy did in this very room before our very eyes last night!' 'I don't so much mind that, once in a way,' remarked Mr Edward; 'but your Clennam, as he thinks proper to call himself, is another thing.' 'He is part of the same thing,' returned Miss Fanny, 'and of a piece with all the rest. He obtruded himself upon us in the first instance. We never wanted him. I always showed him, for one, that I could have dispensed with his company with the greatest pleasure. He then commits that gross outrage upon our feelings, which he never could or would have committed but for the delight he took in exposing us; and then we are to be demeaned for the service of his friends! Why, I don't wonder at this Mr Gowan's conduct towards you. What else was to be expected when he was enjoying our past misfortunes--gloating over them at the moment!' 'Father--Edward--no indeed!' pleaded Little Dorrit. 'Neither Mr nor Mrs Gowan had ever heard our name. They were, and they are, quite ignorant of our history.' 'So much the worse,' retorted Fanny, determined not to admit anything in extenuation, 'for then you have no excuse. If they had known about us, you might have felt yourself called upon to conciliate them. That would have been a weak and ridiculous mistake, but I can respect a mistake, whereas I can't respect a wilful and deliberate abasing of those who should be nearest and dearest to us. No. I can't respect that. I can do nothing but denounce that.' 'I never offend you wilfully, Fanny,' said Little Dorrit, 'though you are so hard with me.' 'Then you should be more careful, Amy,' returned her sister. 'If you do such things by accident, you should be more careful. If I happened to have been born in a peculiar place, and under peculiar circumstances that blunted my knowledge of propriety, I fancy I should think myself bound to consider at every step, "Am I going, ignorantly, to compromise any near and dear relations?" That is what I fancy _I_ should do, if it was _my_ case.' Mr Dorrit now interposed, at once to stop these painful subjects by his authority, and to point their moral by his wisdom. 'My dear,' said he to his younger daughter, 'I beg you to--ha--to say no more. Your sister Fanny expresses herself strongly, but not without considerable reason. You have now a--hum--a great position to support. That great position is not occupied by yourself alone, but by--ha--by me, and--ha hum--by us. Us. Now, it is incumbent upon all people in an exalted position, but it is particularly so on this family, for reasons which I--ha--will not dwell upon, to make themselves respected. To be vigilant in making themselves respected. Dependants, to respect us, must be--ha--kept at a distance and--hum--kept down. Down. Therefore, your not exposing yourself to the remarks of our attendants by appearing to have at any time dispensed with their services and performed them for yourself, is--ha--highly important.' 'Why, who can doubt it?' cried Miss Fanny. 'It's the essence of everything.' 'Fanny,' returned her father, grandiloquently, 'give me leave, my dear. We then come to--ha--to Mr Clennam. I am free to say that I do not, Amy, share your sister's sentiments--that is to say altogether--hum-- altogether--in reference to Mr Clennam. I am content to regard that individual in the light of--ha--generally--a well-behaved person. Hum. A well-behaved person. Nor will I inquire whether Mr Clennam did, at any time, obtrude himself on--ha--my society. He knew my society to be--hum--sought, and his plea might be that he regarded me in the light of a public character. But there were circumstances attending my--ha--slight knowledge of Mr Clennam (it was very slight), which,' here Mr Dorrit became extremely grave and impressive, 'would render it highly indelicate in Mr Clennam to--ha--to seek to renew communication with me or with any member of my family under existing circumstances. If Mr Clennam has sufficient delicacy to perceive the impropriety of any such attempt, I am bound as a responsible gentleman to--ha--defer to that delicacy on his part. If, on the other hand, Mr Clennam has not that delicacy, I cannot for a moment--ha--hold any correspondence with so--hum--coarse a mind. In either case, it would appear that Mr Clennam is put altogether out of the question, and that we have nothing to do with him or he with us. Ha--Mrs General!' The entrance of the lady whom he announced, to take her place at the breakfast-table, terminated the discussion. Shortly afterwards, the courier announced that the valet, and the footman, and the two maids, and the four guides, and the fourteen mules, were in readiness; so the breakfast party went out to the convent door to join the cavalcade. Mr Gowan stood aloof with his cigar and pencil, but Mr Blandois was on the spot to pay his respects to the ladies. When he gallantly pulled off his slouched hat to Little Dorrit, she thought he had even a more sinister look, standing swart and cloaked in the snow, than he had in the fire-light over-night. But, as both her father and her sister received his homage with some favour, she refrained from expressing any distrust of him, lest it should prove to be a new blemish derived from her prison birth. Nevertheless, as they wound down the rugged way while the convent was yet in sight, she more than once looked round, and descried Mr Blandois, backed by the convent smoke which rose straight and high from the chimneys in a golden film, always standing on one jutting point looking down after them. Long after he was a mere black stick in the snow, she felt as though she could yet see that smile of his, that high nose, and those eyes that were too near it. And even after that, when the convent was gone and some light morning clouds veiled the pass below it, the ghastly skeleton arms by the wayside seemed to be all pointing up at him. More treacherous than snow, perhaps, colder at heart, and harder to melt, Blandois of Paris by degrees passed out of her mind, as they came down into the softer regions. Again the sun was warm, again the streams descending from glaciers and snowy caverns were refreshing to drink at, again they came among the pine-trees, the rocky rivulets, the verdant heights and dales, the wooden chalets and rough zigzag fences of Swiss country. Sometimes the way so widened that she and her father could ride abreast. And then to look at him, handsomely clothed in his fur and broadcloths, rich, free, numerously served and attended, his eyes roving far away among the glories of the landscape, no miserable screen before them to darken his sight and cast its shadow on him, was enough. Her uncle was so far rescued from that shadow of old, that he wore the clothes they gave him, and performed some ablutions as a sacrifice to the family credit, and went where he was taken, with a certain patient animal enjoyment, which seemed to express that the air and change did him good. In all other respects, save one, he shone with no light but such as was reflected from his brother. His brother's greatness, wealth, freedom, and grandeur, pleased him without any reference to himself. Silent and retiring, he had no use for speech when he could hear his brother speak; no desire to be waited on, so that the servants devoted themselves to his brother. The only noticeable change he originated in himself, was an alteration in his manner to his younger niece. Every day it refined more and more into a marked respect, very rarely shown by age to youth, and still more rarely susceptible, one would have said, of the fitness with which he invested it. On those occasions when Miss Fanny did declare once for all, he would take the next opportunity of baring his grey head before his younger niece, and of helping her to alight, or handing her to the carriage, or showing her any other attention, with the profoundest deference. Yet it never appeared misplaced or forced, being always heartily simple, spontaneous, and genuine. Neither would he ever consent, even at his brother's request, to be helped to any place before her, or to take precedence of her in anything. So jealous was he of her being respected, that, on this very journey down from the Great Saint Bernard, he took sudden and violent umbrage at the footman's being remiss to hold her stirrup, though standing near when she dismounted; and unspeakably astonished the whole retinue by charging at him on a hard-headed mule, riding him into a corner, and threatening to trample him to death. They were a goodly company, and the Innkeepers all but worshipped them. Wherever they went, their importance preceded them in the person of the courier riding before, to see that the rooms of state were ready. He was the herald of the family procession. The great travelling-carriage came next: containing, inside, Mr Dorrit, Miss Dorrit, Miss Amy Dorrit, and Mrs General; outside, some of the retainers, and (in fine weather) Edward Dorrit, Esquire, for whom the box was reserved. Then came the chariot containing Frederick Dorrit, Esquire, and an empty place occupied by Edward Dorrit, Esquire, in wet weather. Then came the fourgon with the rest of the retainers, the heavy baggage, and as much as it could carry of the mud and dust which the other vehicles left behind. These equipages adorned the yard of the hotel at Martigny, on the return of the family from their mountain excursion. Other vehicles were there, much company being on the road, from the patched Italian Vettura--like the body of a swing from an English fair put upon a wooden tray on wheels, and having another wooden tray without wheels put atop of it--to the trim English carriage. But there was another adornment of the hotel which Mr Dorrit had not bargained for. Two strange travellers embellished one of his rooms. The Innkeeper, hat in hand in the yard, swore to the courier that he was blighted, that he was desolated, that he was profoundly afflicted, that he was the most miserable and unfortunate of beasts, that he had the head of a wooden pig. He ought never to have made the concession, he said, but the very genteel lady had so passionately prayed him for the accommodation of that room to dine in, only for a little half-hour, that he had been vanquished. The little half-hour was expired, the lady and gentleman were taking their little dessert and half-cup of coffee, the note was paid, the horses were ordered, they would depart immediately; but, owing to an unhappy destiny and the curse of Heaven, they were not yet gone. Nothing could exceed Mr Dorrit's indignation, as he turned at the foot of the staircase on hearing these apologies. He felt that the family dignity was struck at by an assassin's hand. He had a sense of his dignity, which was of the most exquisite nature. He could detect a design upon it when nobody else had any perception of the fact. His life was made an agony by the number of fine scalpels that he felt to be incessantly engaged in dissecting his dignity. 'Is it possible, sir,' said Mr Dorrit, reddening excessively, 'that you have--ha--had the audacity to place one of my rooms at the disposition of any other person?' Thousands of pardons! It was the host's profound misfortune to have been overcome by that too genteel lady. He besought Monseigneur not to enrage himself. He threw himself on Monseigneur for clemency. If Monseigneur would have the distinguished goodness to occupy the other salon especially reserved for him, for but five minutes, all would go well. 'No, sir,' said Mr Dorrit. 'I will not occupy any salon. I will leave your house without eating or drinking, or setting foot in it. How do you dare to act like this? Who am I that you--ha--separate me from other gentlemen?' Alas! The host called all the universe to witness that Monseigneur was the most amiable of the whole body of nobility, the most important, the most estimable, the most honoured. If he separated Monseigneur from others, it was only because he was more distinguished, more cherished, more generous, more renowned. 'Don't tell me so, sir,' returned Mr Dorrit, in a mighty heat. 'You have affronted me. You have heaped insults upon me. How dare you? Explain yourself.' Ah, just Heaven, then, how could the host explain himself when he had nothing more to explain; when he had only to apologise, and confide himself to the so well-known magnanimity of Monseigneur! 'I tell you, sir,' said Mr Dorrit, panting with anger, 'that you separate me--ha--from other gentlemen; that you make distinctions between me and other gentlemen of fortune and station. I demand of you, why? I wish to know on--ha--what authority, on whose authority. Reply sir. Explain. Answer why.' Permit the landlord humbly to submit to Monsieur the Courier then, that Monseigneur, ordinarily so gracious, enraged himself without cause. There was no why. Monsieur the Courier would represent to Monseigneur, that he deceived himself in suspecting that there was any why, but the why his devoted servant had already had the honour to present to him. The very genteel lady-- 'Silence!' cried Mr Dorrit. 'Hold your tongue! I will hear no more of the very genteel lady; I will hear no more of you. Look at this family--my family--a family more genteel than any lady. You have treated this family with disrespect; you have been insolent to this family. I'll ruin you. Ha--send for the horses, pack the carriages, I'll not set foot in this man's house again!' No one had interfered in the dispute, which was beyond the French colloquial powers of Edward Dorrit, Esquire, and scarcely within the province of the ladies. Miss Fanny, however, now supported her father with great bitterness; declaring, in her native tongue, that it was quite clear there was something special in this man's impertinence; and that she considered it important that he should be, by some means, forced to give up his authority for making distinctions between that family and other wealthy families. What the reasons of his presumption could be, she was at a loss to imagine; but reasons he must have, and they ought to be torn from him. All the guides, mule-drivers, and idlers in the yard, had made themselves parties to the angry conference, and were much impressed by the courier's now bestirring himself to get the carriages out. With the aid of some dozen people to each wheel, this was done at a great cost of noise; and then the loading was proceeded with, pending the arrival of the horses from the post-house. But the very genteel lady's English chariot being already horsed and at the inn-door, the landlord had slipped up-stairs to represent his hard case. This was notified to the yard by his now coming down the staircase in attendance on the gentleman and the lady, and by his pointing out the offended majesty of Mr Dorrit to them with a significant motion of his hand. 'Beg your pardon,' said the gentleman, detaching himself from the lady, and coming forward. 'I am a man of few words and a bad hand at an explanation--but lady here is extremely anxious that there should be no Row. Lady--a mother of mine, in point of fact--wishes me to say that she hopes no Row.' Mr Dorrit, still panting under his injury, saluted the gentleman, and saluted the lady, in a distant, final, and invincible manner. 'No, but really--here, old feller; you!' This was the gentleman's way of appealing to Edward Dorrit, Esquire, on whom he pounced as a great and providential relief. 'Let you and I try to make this all right. Lady so very much wishes no Row.' Edward Dorrit, Esquire, led a little apart by the button, assumed a diplomatic expression of countenance in replying, 'Why you must confess, that when you bespeak a lot of rooms beforehand, and they belong to you, it's not pleasant to find other people in 'em.' 'No,' said the other, 'I know it isn't. I admit it. Still, let you and I try to make it all right, and avoid Row. The fault is not this chap's at all, but my mother's. Being a remarkably fine woman with no bigodd nonsense about her--well educated, too--she was too many for this chap. Regularly pocketed him.' 'If that's the case--' Edward Dorrit, Esquire, began. 'Assure you 'pon my soul 'tis the case. Consequently,' said the other gentleman, retiring on his main position, 'why Row?' 'Edmund,' said the lady from the doorway, 'I hope you have explained, or are explaining, to the satisfaction of this gentleman and his family that the civil landlord is not to blame?' 'Assure you, ma'am,' returned Edmund, 'perfectly paralysing myself with trying it on.' He then looked steadfastly at Edward Dorrit, Esquire, for some seconds, and suddenly added, in a burst of confidence, 'Old feller! _Is_ it all right?' 'I don't know, after all,' said the lady, gracefully advancing a step or two towards Mr Dorrit, 'but that I had better say myself, at once, that I assured this good man I took all the consequences on myself of occupying one of a stranger's suite of rooms during his absence, for just as much (or as little) time as I could dine in. I had no idea the rightful owner would come back so soon, nor had I any idea that he had come back, or I should have hastened to make restoration of my ill-gotten chamber, and to have offered my explanation and apology. I trust in saying this--' For a moment the lady, with a glass at her eye, stood transfixed and speechless before the two Miss Dorrits. At the same moment, Miss Fanny, in the foreground of a grand pictorial composition, formed by the family, the family equipages, and the family servants, held her sister tight under one arm to detain her on the spot, and with the other arm fanned herself with a distinguished air, and negligently surveyed the lady from head to foot. The lady, recovering herself quickly--for it was Mrs Merdle and she was not easily dashed--went on to add that she trusted in saying this, she apologised for her boldness, and restored this well-behaved landlord to the favour that was so very valuable to him. Mr Dorrit, on the altar of whose dignity all this was incense, made a gracious reply; and said that his people should--ha--countermand his horses, and he would--hum--overlook what he had at first supposed to be an affront, but now regarded as an honour. Upon this the bosom bent to him; and its owner, with a wonderful command of feature, addressed a winning smile of adieu to the two sisters, as young ladies of fortune in whose favour she was much prepossessed, and whom she had never had the gratification of seeing before. Not so, however, Mr Sparkler. This gentleman, becoming transfixed at the same moment as his lady-mother, could not by any means unfix himself again, but stood stiffly staring at the whole composition with Miss Fanny in the Foreground. On his mother saying, 'Edmund, we are quite ready; will you give me your arm?' he seemed, by the motion of his lips, to reply with some remark comprehending the form of words in which his shining talents found the most frequent utterance, but he relaxed no muscle. So fixed was his figure, that it would have been matter of some difficulty to bend him sufficiently to get him in the carriage-door, if he had not received the timely assistance of a maternal pull from within. He was no sooner within than the pad of the little window in the back of the chariot disappeared, and his eye usurped its place. There it remained as long as so small an object was discernible, and probably much longer, staring (as though something inexpressibly surprising should happen to a codfish) like an ill-executed eye in a large locket. This encounter was so highly agreeable to Miss Fanny, and gave her so much to think of with triumph afterwards, that it softened her asperities exceedingly. When the procession was again in motion next day, she occupied her place in it with a new gaiety; and showed such a flow of spirits indeed, that Mrs General looked rather surprised. Little Dorrit was glad to be found no fault with, and to see that Fanny was pleased; but her part in the procession was a musing part, and a quiet one. Sitting opposite her father in the travelling-carriage, and recalling the old Marshalsea room, her present existence was a dream. All that she saw was new and wonderful, but it was not real; it seemed to her as if those visions of mountains and picturesque countries might melt away at any moment, and the carriage, turning some abrupt corner, bring up with a jolt at the old Marshalsea gate. To have no work to do was strange, but not half so strange as having glided into a corner where she had no one to think for, nothing to plan and contrive, no cares of others to load herself with. Strange as that was, it was far stranger yet to find a space between herself and her father, where others occupied themselves in taking care of him, and where she was never expected to be. At first, this was so much more unlike her old experience than even the mountains themselves, that she had been unable to resign herself to it, and had tried to retain her old place about him. But he had spoken to her alone, and had said that people--ha--people in an exalted position, my dear, must scrupulously exact respect from their dependents; and that for her, his daughter, Miss Amy Dorrit, of the sole remaining branch of the Dorrits of Dorsetshire, to be known to--hum--to occupy herself in fulfilling the functions of--ha hum--a valet, would be incompatible with that respect. Therefore, my dear, he--ha--he laid his parental injunctions upon her, to remember that she was a lady, who had now to conduct herself with--hum--a proper pride, and to preserve the rank of a lady; and consequently he requested her to abstain from doing what would occasion--ha--unpleasant and derogatory remarks. She had obeyed without a murmur. Thus it had been brought about that she now sat in her corner of the luxurious carriage with her little patient hands folded before her, quite displaced even from the last point of the old standing ground in life on which her feet had lingered. It was from this position that all she saw appeared unreal; the more surprising the scenes, the more they resembled the unreality of her own inner life as she went through its vacant places all day long. The gorges of the Simplon, its enormous depths and thundering waterfalls, the wonderful road, the points of danger where a loose wheel or a faltering horse would have been destruction, the descent into Italy, the opening of that beautiful land as the rugged mountain-chasm widened and let them out from a gloomy and dark imprisonment--all a dream--only the old mean Marshalsea a reality. Nay, even the old mean Marshalsea was shaken to its foundations when she pictured it without her father. She could scarcely believe that the prisoners were still lingering in the close yard, that the mean rooms were still every one tenanted, and that the turnkey still stood in the Lodge letting people in and out, all just as she well knew it to be. With a remembrance of her father's old life in prison hanging about her like the burden of a sorrowful tune, Little Dorrit would wake from a dream of her birth-place into a whole day's dream. The painted room in which she awoke, often a humbled state-chamber in a dilapidated palace, would begin it; with its wild red autumnal vine-leaves overhanging the glass, its orange-trees on the cracked white terrace outside the window, a group of monks and peasants in the little street below, misery and magnificence wrestling with each other upon every rood of ground in the prospect, no matter how widely diversified, and misery throwing magnificence with the strength of fate. To this would succeed a labyrinth of bare passages and pillared galleries, with the family procession already preparing in the quadrangle below, through the carriages and luggage being brought together by the servants for the day's journey. Then breakfast in another painted chamber, damp-stained and of desolate proportions; and then the departure, which, to her timidity and sense of not being grand enough for her place in the ceremonies, was always an uneasy thing. For then the courier (who himself would have been a foreign gentleman of high mark in the Marshalsea) would present himself to report that all was ready; and then her father's valet would pompously induct him into his travelling-cloak; and then Fanny's maid, and her own maid (who was a weight on Little Dorrit's mind--absolutely made her cry at first, she knew so little what to do with her), would be in attendance; and then her brother's man would complete his master's equipment; and then her father would give his arm to Mrs General, and her uncle would give his to her, and, escorted by the landlord and Inn servants, they would swoop down-stairs. There, a crowd would be collected to see them enter their carriages, which, amidst much bowing, and begging, and prancing, and lashing, and clattering, they would do; and so they would be driven madly through narrow unsavoury streets, and jerked out at the town gate. Among the day's unrealities would be roads where the bright red vines were looped and garlanded together on trees for many miles; woods of olives; white villages and towns on hill-sides, lovely without, but frightful in their dirt and poverty within; crosses by the way; deep blue lakes with fairy islands, and clustering boats with awnings of bright colours and sails of beautiful forms; vast piles of building mouldering to dust; hanging-gardens where the weeds had grown so strong that their stems, like wedges driven home, had split the arch and rent the wall; stone-terraced lanes, with the lizards running into and out of every chink; beggars of all sorts everywhere: pitiful, picturesque, hungry, merry; children beggars and aged beggars. Often at posting-houses and other halting places, these miserable creatures would appear to her the only realities of the day; and many a time, when the money she had brought to give them was all given away, she would sit with her folded hands, thoughtfully looking after some diminutive girl leading her grey father, as if the sight reminded her of something in the days that were gone. Again, there would be places where they stayed the week together in splendid rooms, had banquets every day, rode out among heaps of wonders, walked through miles of palaces, and rested in dark corners of great churches; where there were winking lamps of gold and silver among pillars and arches, kneeling figures dotted about at confessionals and on the pavements; where there was the mist and scent of incense; where there were pictures, fantastic images, gaudy altars, great heights and distances, all softly lighted through stained glass, and the massive curtains that hung in the doorways. From these cities they would go on again, by the roads of vines and olives, through squalid villages, where there was not a hovel without a gap in its filthy walls, not a window with a whole inch of glass or paper; where there seemed to be nothing to support life, nothing to eat, nothing to make, nothing to grow, nothing to hope, nothing to do but die. Again they would come to whole towns of palaces, whose proper inmates were all banished, and which were all changed into barracks: troops of idle soldiers leaning out of the state windows, where their accoutrements hung drying on the marble architecture, and showing to the mind like hosts of rats who were (happily) eating away the props of the edifices that supported them, and must soon, with them, be smashed on the heads of the other swarms of soldiers and the swarms of priests, and the swarms of spies, who were all the ill-looking population left to be ruined, in the streets below. Through such scenes, the family procession moved on to Venice. And here it dispersed for a time, as they were to live in Venice some few months in a palace (itself six times as big as the whole Marshalsea) on the Grand Canal. In this crowning unreality, where all the streets were paved with water, and where the deathlike stillness of the days and nights was broken by no sound but the softened ringing of church-bells, the rippling of the current, and the cry of the gondoliers turning the corners of the flowing streets, Little Dorrit, quite lost by her task being done, sat down to muse. The family began a gay life, went here and there, and turned night into day; but she was timid of joining in their gaieties, and only asked leave to be left alone. Sometimes she would step into one of the gondolas that were always kept in waiting, moored to painted posts at the door--when she could escape from the attendance of that oppressive maid, who was her mistress, and a very hard one--and would be taken all over the strange city. Social people in other gondolas began to ask each other who the little solitary girl was whom they passed, sitting in her boat with folded hands, looking so pensively and wonderingly about her. Never thinking that it would be worth anybody's while to notice her or her doings, Little Dorrit, in her quiet, scared, lost manner, went about the city none the less. But her favourite station was the balcony of her own room, overhanging the canal, with other balconies below, and none above. It was of massive stone darkened by ages, built in a wild fancy which came from the East to that collection of wild fancies; and Little Dorrit was little indeed, leaning on the broad-cushioned ledge, and looking over. As she liked no place of an evening half so well, she soon began to be watched for, and many eyes in passing gondolas were raised, and many people said, There was the little figure of the English girl who was always alone. Such people were not realities to the little figure of the English girl; such people were all unknown to her. She would watch the sunset, in its long low lines of purple and red, and its burning flush high up into the sky: so glowing on the buildings, and so lightening their structure, that it made them look as if their strong walls were transparent, and they shone from within. She would watch those glories expire; and then, after looking at the black gondolas underneath, taking guests to music and dancing, would raise her eyes to the shining stars. Was there no party of her own, in other times, on which the stars had shone? To think of that old gate now! She would think of that old gate, and of herself sitting at it in the dead of the night, pillowing Maggy's head; and of other places and of other scenes associated with those different times. And then she would lean upon her balcony, and look over at the water, as though they all lay underneath it. When she got to that, she would musingly watch its running, as if, in the general vision, it might run dry, and show her the prison again, and herself, and the old room, and the old inmates, and the old visitors: all lasting realities that had never changed.

Summary: The next morning the Dorrits and their huge retinue of servants pack up to go down the mountains into Italy. Amy tells Tip that Pet Gowan is doing better, and he worries that she's been trying to take care of her - which is low-class and something only servants are supposed to do. Fanny overhears this exchange and loses it. She figures out that Pet must know Arthur, who apparently did something she finds super-offensive in between the novel's two books. It's not clear what this thing Arthur did is, but Fanny writes him off entirely. Dorrit is less angry about Arthur but tells Amy she is just not doing enough to preserve their family dignity. Poor Amy. No one wants to hang out with Prisony McPrisonson. The only person who is nice to her is her uncle Frederick, who goes out of his way to make sure she is well taken care of and that everyone defers to her. Too bad he's a half-senile old man. After some traveling, the Dorrits get to Venice. When they arrive at their hotel, the innkeeper tells them that a lady and gentleman are using their dining room and will be out in a minute. Dorrit flips his lid and starts yelling at the innkeeper - why is he being treated as less important than the lady? The poor guy doesn't know what to say, and Dorrit threatens to leave the hotel, until out of the dining room pops... Edmund Sparkler! He's all apologies and stuff, and his mom Mrs. Merdle is not too far behind with some more apologies. In the middle of apologizing, she recognizes Fanny and Amy. But Mrs. Merdle is a lady of the world, so she plays it off like she's never met either of them before. Dorrit is satisfied with the apologies, and Mrs. Merdle and Edmund Sparkler go off in the carriage, with Edmund never taking his eyes off Fanny. Fanny is totally psyched by this encounter and is a in a good mood for a while afterwards. Amy, meanwhile, is living in kind of a dream world. They go around Venice and everything seems dreamy and strange to her - the canals and the museums and all the people. All she does is think about the prison and wonder how it's possible that it still exists. It's also very stressful for her to no longer be taking physical care of her father's needs and to be replaced by servants. Looks like Amy is going to have to find a new identity!

booksum

en

Summarize: FIELD OF THE INVENTION The present invention relates generally to a system or apparatus for thermally treating medical instruments. More particularly, but not exclusively, the invention relates towards an improved system and apparatus for covering a thermal medical treatment apparatus and for supporting a medical instrument being warmed by the thermal medical treatment apparatus. BACKGROUND OF THE INVENTION Surgical scopes (e.g., laparoscopes, endoscopes, arthroscopes, etc.) are used in corrective medical procedures, as well as in medical procedures that image interior viscera such as surfaces of the stomach, small intestines, and colon. The use of surgical scopes permits a surgeon to view a patient body interior with a minimal amount of cutting of patient tissue. The surgical scopes may be warmed prior to use, where scope optics must remain dry to protect those optics and prevent distortion of the image. The scopes are warmed for several reasons, including enhancing image results, preventing infections, and maintaining normothermia. For example, a scope that is not warmed prior to being inserted into a patient body may fog due to differences between the body temperature and scope temperature, thereby impeding or distorting the resulting image. Further, scopes may be warmed to minimize trauma caused to tissue in response to insertion of the scope into the patient body. The trauma basically results from the temperature difference between the scope and the tissue. Inserting a hot or cold scope may damage tissue, thereby leading to infections. Inserting a cold scope may also lower core body temperature, thereby leading to hypothermia and compromising patient safety. Therefore, thermal warming systems or devices may warm the scopes. The thermal devices generally include a body or cabinet, and a basin or recess positioned within a top surface of the body. The basin is configured to contain and thermally treat a liquid bath. The medical instrument, or scope, is then placed in the thermally treated bath to raise the temperature of the insertion end of the scope to the desired temperature prior to inserting the scope into the patient. The opposite end, or optical end, of the scope is positioned outside of the bath so as to not damage the optics, which can be expensive to replace. However, it should also be appreciated that the scopes and other medical instruments and devices must be sterile, so as to reduce the chance of infecting a patient during surgery or other procedures. Therefore, sterilized plastic sheets have been placed over the thermal warming systems, including the basis, to reduce the risk of infection. The liquid is added directly on the sheet in the basin, with additional parts of the sheet hanging over and around the device. The medical scopes are often longer than the length of the basin and distance to an edge of the warming device. Thus, when the end of the scope is being warmed in the basin, the opposite end extends at an angle beyond the edge of the device, which increases the chance that the scope can fall out of the basin. Doing so can introduce the insertion end of the scope to potentially harmful bacteria, and can also damage the optic end of the scope, which, as noted above, can be rather expensive. Methods have been taken by medical staff to reduce the chance that the scope will fall out of or off of the thermal device. For example, it is not uncommon for a surgical team to move a wheeled shelf adjacent the wheeled thermal device, such that the optic end of the scope can rest on the shelf, with the optic end elevated to ensure that the insertion end remains in the thermally treated liquid solution. However, doing so reduces the amount of shelf space available in the surgery room, and also introduces additional risks to the scope. An otherwise useful shelf must be used to support the scope, or an otherwise unneeded shelf must be used, which reduces the amount of space in the room. As both the second shelf and the thermal device are on wheels, they can be easily moved, even when not purposefully moved. Moving the second shelf, either purposefully or accidentally, will move the support of the scope, which can cause the scope to fall from the thermal device, introducing the problems discussed above. Therefore, there is a need in the art for an apparatus and system that can be used with a thermal warming medical device or apparatus that covers the device to ensure a sterilized medical room. There is also a need for a medical drape or cover that includes a scope support member housed within to support a scope during warming of the scope in the medical device. Furthermore, there is a need for a shelf that can be attached to the medical device to increase the amount of usable space, while minimizing intrusion into the medical room space. The shelf can also be used to support the support member of the drape without having to use a separate wheeled shelf in the room. SUMMARY OF THE INVENTION Therefore, it is principal object, feature, and/or advantage of the present invention to provide an apparatus that overcomes the deficiencies in the art. It is another object, feature, and/or advantage of the present invention to provide a thermal device drape that includes a support member for supporting a medical instrument that needs warmed or otherwise sterilized. It is still another object, feature, and/or advantage of the present invention to provide a support member within a drape that includes various shapes to prevent medical instruments from rolling off the support. It is a further object, feature, and/or advantage of the present invention to provide a drape and system for a warming medical device that includes a shelf for supporting a medical instrument support member for the warming device. It is still a further object, feature, and/or advantage of the present invention to provide a system that includes a cord hook for redirecting and routing a power cord for a warming medical device. It is yet a further object, feature, and/or advantage of the present invention to provide a system including a drape, support member, extension shelf, and hook to provide stability for a warming medical device while not reducing space in a medical room. These and/or other objects, features, and advantages of the present invention will be apparent to those skilled in the art. The present invention is not to be limited to or by these objects, features and advantages. No single embodiment need provide each and every object, feature, or advantage. According to one aspect of the present invention, a drape for use with a thermal medical treatment apparatus having a body including a recessed basin in a top surface is provided. The drape includes a sheet for covering the apparatus basin and body and hanging from the apparatus; and a support member housed in the sheet for supporting a medical device adjacent the basin. According to another aspect of the present invention, a drape for use with a thermal medical treatment apparatus having a body including a recessed basin in a top surface is provided. The drape includes a first sheet for covering the apparatus basin and body and hanging from the apparatus; a second sheet attached to the first sheet to form a pocket therebetween; and a support member positioned in the pocket of the first and second sheets, the support member configured to support a medical device adjacent the basin. According to yet another aspect of the present invention, a system for covering a thermal medical treatment apparatus having a body including a recessed basin in a top surface is provided. The system includes an extension shelf attached to the apparatus body adjacent the basin; a sheet covering the extension shelf, apparatus basin, and apparatus body and hanging therefrom; and a support member housed in the sheet and positioned on the extension shelf for supporting a medical device adjacent the basin. The invention includes a pillow drape with extension shelf and power cord hook to offer improved stability of scopes during warming. The extension shelf is designed to offer increased working surface area of the warmer while minimizing intrusion into the operating room space. The power cord hook allows for power cord storage and convenient change of orientation relative to the sterile field. The pillow drape incorporates a foam holder to provide stable scope positioning during warming. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view of a medical warming device with a drape system attached and covering the same according to the present invention. FIG. 2 is a side elevation view of the device and system of FIG. 1. FIG. 3 is a front elevation view of the device and system of FIG. 1. FIG. 4 is an exploded view of the device and system of FIG. 1. FIG. 5 is an exploded view of a pillow drape for use with the drape system according to an embodiment of the present invention. FIG. 6 is a perspective view of an extension shelf for use with the drape system according to an embodiment of the present invention. FIG. 7 is a perspective view of a cord hook for use with the drape system according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a perspective view of a medical warming device with a drape system attached thereto and covering the same, according to the present invention. As shown in FIG. 1, a medical system 10 includes, in part, a pillow drape 12 and a warming device 40. The warming device 40 is a medical apparatus used to warm scopes 58, such as surgical scopes (e.g., endoscopes or the like). The scopes 58 generally include an insertion end 60 and an optic end 62, which contains a camera or other optical device. However, as the insertion end 60 will be inserted into a patient, it is necessary to warm the insertion end 60 prior to insertion. Therefore, the scope 58 is placed in a liquid 48 contained in a basin 46 formed in a top surface 44 of the body 42 of the warming device 40. The insertion end 60 of the scope 58 can be warmed to a desired temperature prior to inserting the scope in the patient during surgery or other medical procedures. Furthermore, the warming device 40 may be attached to a stand 52 including a plurality of wheels 54 to move the warming device 40 as needed in the surgical or medical room. In order to ensure a sterile environment in the surgical room, a sterilized pillow drape 12 is included to cover the warming device 40, including the basin 46. As shown in FIG. 1, the pillow drape 12 covers the warming device 40, and the liquid 48 is added on top of the sterilized pillow drape 12, and the scope 58 is then included in the warming device 40 to warm as needed. An embodiment of the pillow drape 12 is shown in FIGS. 1-4. The pillow drape 12 includes generally a sheet 14, which may be a sterilized polyurethane sheet. It is contemplated that the sheet 14 be clear or be colored as needed or desired. The pillow drape 12 also includes a first section 16 and a second section 18. The first section 16 includes generally the sheet 14, which may also be known as a first sheet. The second section 18 includes a cover portion 20 defining a pocket 24 between the first and second sections. Housed within the pocket 24 is a support member 32. The support member 32, as shown in FIG. 1, can be used to support an end, shown to be the optic end 62, of the scope 58 in the warming device 40. The support member 32 includes a surface 34 and first and second walls 36, 38 extending generally upwards from the first surface 34. The first and second walls 36, 38 aid in preventing the scope 58 from rolling off of the support member and warming device. However, it should be contemplated that the exact shape of the support member 32 need not be that shown in FIGS. 1-3, and that generally any shape may be used to aid in supporting the scope 58 in the warming device 40. The support member 32 may comprise generally any material capable of supporting the scope 58 in the warming device 40. However, according to one embodiment, a foam material is used. The density of the foam may vary according to the type of scope used, as well as other factors. The present invention contemplates that any density be used, and that the invention is not to be limited to any such densities. Therefore, the support member 32 may be designed and cut out of a bulk piece of foam to the desired shape for the pillow drape 12. The size of the support member 32 may be adjusted and configured as necessary. For example, in some situations, multiple scopes will be needed to be warmed in the warming device at the same time. Therefore, the present invention contemplates that a generally wider support member 32 (wider surface 34 ) be used to support the plurality of scopes. Furthermore, it is contemplated that the support member 32 of the present invention comprises a generally pliable or deformable material such that the resting of the scope 58 on the support member 32 will cause a surface of the support member to slightly deform. The deformation of the support member 32 due to the weight of the scope will further aid in preventing the accidental falling of the scope from the warming device 40, while also providing a surface that will not scratch or otherwise damage the scope 58. Furthermore, as shown in FIG. 1, the support member 32 will aid in raising an end of the scope 58 such that the insertion end 60 of the scope 58 will be in the warming liquid at all times. This will aid in ensuring that the scope is warmed to the ideal or required temperature. As is also shown in FIGS. 1-4, an extension shelf 64 can be used with the present invention. The extension shelf 64 includes top and bottom surfaces 66, 68, and a support or attachment means 72 for attaching the shelf to the body 42 of the warming device 40. The support shelf may simply be screwed or otherwise adhered to body 42 of the warming device 40. The extension shelf 64 provides increased area to allow the support member to rest thereon during use of the pillow drape 12 of the present invention. As noted, the amount of space in a surgical or medical room is limited. Therefore, the extension shelf provides additional space for the warming device 40 and pillow drape 12, while not reducing the usable space in the medical room. The extension shelf 64 allows for an easy attachment to the warming device to provide support for the pillow drape 12, including the support member 32. Thus, as shown in FIG. 4, the support member 32 may rest or be positioned on the extension shelf 64 before placing the scope 58 in the warming device 40. Furthermore, while the extension shelf 64 of the present invention is shown to extend generally only at one side or end of the warming device 40, it is contemplated by the present invention that the extension shelf may be any size and configuration needed. For example, the extension shelf could extend from an end and along a side of the warming device 40 to provide additional space for medical personnel to place medical tools and other devices. The extension shelf may also be smaller, if the warming device 40 is smaller or support member 32 is smaller. This may be beneficial when the amount of available space is even less than ideal. According to one embodiment of the present invention, the extension shelf 64 is positioned opposite the controls 50 of the warming device 40. Doing so aids in keeping the raised (optic) end 62 of the scope 58 away from the controls 50, such that adjusting the controls 50 will include a lower or reduced risk of altering the position of the scope 48, or accidentally knocking the scope 48 off of the device 40. However, as medical devices, such as the warming device 40 shown in the figures, include varying configurations and placements of controls and other information, the location of the extension shelf 64 and support member 32 need not be in the exact configuration shown in the Figures, and can be positioned adjacent the device to best increase space and reduce damage to the equipment. Also shown in the Figures is a cord hook 74 attached to the warming device 40. As shown best in FIG. 2, a power cord 56 for the warming device 40 extends from an underside of the warming device 40. As the extension shelf 64 and support member 32 of the pillow drape 12 is positioned on the same side as the power cord 56, it may be desired to route the power cord 56 to the opposite side such that it can be out of the way for medical personnel. Therefore, the cord hook 74 can be attached to the underside of the warming device 40, and the power cord 56 can be routed under the warming device and through a router 78 of the hook 74 to route the cord away from the support member 32 side of the medical system 10. The cord hook 74 can be attached in any manner to the warming device 40. For example, the cord hook 74 can be screwed, glued, taped, or otherwise adhered to the warming device 40. In addition, while the cord hook 74 is shown to be attached to the underside of the warming device 40, it is contemplated that the hook can be placed or attached generally anywhere to the warming device 40 such that it is able to support the power cord 56. However, the location of the cord hook 74 as shown in the Figures is positioned such that the hook can be attached to the warming device using existing screw holes. FIG. 5 is an exploded view of the pillow drape 12 for use with the system 10 according to an embodiment of the present invention. As mentioned above, the pillow drape 12 can be comprised of a first section 16, including a sheet 14, and a second section 18, including a cover portion 20. As noted previously, the first section 16 or sheet 14 comprises a sterilized plastic, such as polyurethane. The sheet 14 may be opaque or also may be transparent. Slits 28 can be placed into the sheet at the location of the support member and second section 18, and can be used to aide in sterilizing the pillow drape 12, as will be described below. The support member 32 and second section 18 (cover section 20 ) are then positioned or attached to the sheet at the slits 28. As shown in FIG. 5, the second section 18 may include a cover portion 20 and a flange-type portion 22 to define a pocket 24 for housing the support member 32. While the cover portion 20 or pocket 24 shown in FIG. 5 is shown to be the same shape as a support member 32, the shape is for purposes of understanding, and is not to be limiting to the present invention. The cover portion 20 forming the pocket 24 can be any shape, including a simple dome shape that is able to conform to the shape of the support member 32. Furthermore, the second section 18 of the pillow drape 12 will also be formed of a sterilized plastic, such as polyurethane. To form the pillow drape 12 of the present invention, the following method can be used. The desired size and shape of the support member 32, including the first surface 34, and first and second side walls 36, 38 is decided. The support member is then formed into the said shape, such as by cutting a piece of foam into the desired shape of the support member 32. A second section or sheet 18 of sterilized polyurethane can be positioned through a hole on a forming board with a flange portion 22 around the edge of the hole to aid in holding the second section in place. The support member 32 is then inserted into the now formed pocket 24 in the second section 18 in an upside down orientation. The first section 16 can then be placed on top of the support board with the slits 28 positioned adjacent the support member 32 and within the opening of the second section. The first and second sections 16, 18 can then be sealed to one another, such as melting, adhering, or the like, to connect the first and second portions to form a single pillow drape 12. The now formed pillow drape 12, including the first section 16, second section 20, and support member 32 can then be sterilized according to known sterilization processes. The location of the slits 28 adjacent to the support member will allow the sterilization process to sterilize the support member 32 within the pocket 24 between the first and second sheets 16, 18, along with the rest of the pillow drape 12. The pillow drape 12 can then be packaged and sent to be used with a warming device 40. In addition, the extension shelf 64 and cord hook 74 can be included with the pillow drape 12 to make the medical system 10 as shown in FIG. 1. In addition, the pillow drape 12 can be formed in other manners as well. For example, the support member 32 could be molded or formed of a thicker piece of plastic, such a polyurethane, to include the support member 32 formed integrally with the first section 16 of the pillow drape 12. Thus, the integrally formed support member 32 would not be a separate piece from the rest of the pillow drape/sheet. Furthermore, the support member 32 could be formed of a material, such as a rubber or plastic, which could be sealed directly to the first sheet 16 of the pillow drape 12. Thus, this would eliminate the need of a second section 18 including a pocket 24. Furthermore, as noted previously, the size and configuration of the support member 32 can dictate the size of the pocket 24 between the first and second sheets 16, 18. The present invention is not to be limited to the configurations shown in the Figures, as well as the exact method described above. FIG. 6 is a perspective view of the cord hook 74 according to an embodiment of the present invention. As noted, the cord hook 74 includes a hook attachment portion 76, and a router portion 78. The attachment portion 76 is used to attach the cord hook 74 to the body 42 of the warming device 40. The router portion 78 is used to hold a route the power cord 56 of the warming device 40. The configuration of the cord hook 74 is not to be limited by that shown in FIG. 6. For example, the router portion 78 may include an enclosed circle, or the addition of additional or a plurality of hooks to allow for the cord hook 74 to route a plurality of power cords for the warming device and any other medical device. Furthermore, as noted previously, while FIG. 6 shows that the cord hook 74 can be attached. by screws at the attachment portion 76 to the warming device 40, it is contemplated that the cord hook be attached to the warming device via other means. For example, the cord hook 74 could be attached via tape, via glue, via VELCRO®(hook and loops), or the like. The cord hook may comprise generally any rigid material, such as, but not limited to, metal, plastic, rubber, or the like. FIG. 7 is a perspective view of an embodiment of the extension shelf 64 according to the present invention. The extension shelf 64 shown in FIG. 7 includes a top surface 66 and an opposite bottom surface 68. The top surface is used to support and hold items in the medical room, such as the support member 32 of the pillow drape 12. However, the extension shelf could also be used to hold other medical instruments, such as scopes and other tools. Thus, the size and shape of the extension shelf can be configured to that configured to any configuration as needed. For example, as noted above, it is contemplated that the shelf extends along one end of the warming device, an end in a side of the warming device, an end in both sides of the warming device, or generally around all sides and ends of the warming device. The present invention is not to be limited by the configurations shown in the Figures. Also shown in FIG. 7 is a plurality of shelf lips 70 surrounding at least a portion of the extension shelf 64. The shelf lip 70 is included to aid in preventing items from falling off the extension shelf. Finally, a shelf support 72 is shown in FIG. 7. The shelf support 72 may also be known as a shelf attachment means. Thus, it is contemplated that the shelf 64 may be attached to the warming device via the shelf support or attachment means 72. As shown in FIG. 7, the shelf can be screwed to the body 42 of the warming device 40. However, it is contemplated that other means will be used to attach the shelf to the warming device 40, such as, but not limited to, screws, VELCRO® (hook and loops), glue, adhesive, tape, or the like. The invention is not to be limited to the exact configuration and attachment means shown and described. The foregoing description has been presented for purposes of illustration and description, and it not intended to be an exhaustive list or to limit the invention to precise forms disclosed. It is contemplated that other alternative processes obvious to those skilled in the art are considered to be included in the invention. The descriptions are merely examples of embodiments. For example, the materials of the various components described may be configured as required. For example, while the sheet has been described as being polyurethane, and the support member as a foam, it is contemplated that the sheet may be any other sterilized material or material used in the medical industry. The support member 32 may also be any material capable of supporting a medical instrument. Furthermore, the size of the pillow drape, including the excess 30 of the sheets, the size and configuration of the support members 32, as well as the size and configuration of the extension shelf may be adjusted as required or desired according to the use and need in the operating or other medical room. It is understood that any other modifications, substitutions, and/or additions may be made, which are within the intended spirit and scope of the invention. From the foregoing, it can be seen that present invention accomplishes at least all of the stated objectives.

Summary: A pillow drape for use with a thermal medical treatment apparatus or device is provided. The drape includes a sterilized sheet for covering the warming medical device that covers the device body and a basin that is used to store a warming liquid. The pillow drape includes an enclosure having a support member therein to support a medical instrument, such as a surgical scope, during warming of the scope. The support member can be housed between first and second sections of the pillow drape sheet in a pocket portion. The support member can be positioned on an extension shelf added to the warming device to provide additional storage room while not taking up room in the surgical or medical room. Additionally, a cord hook can be attached to the medical device to aid in routing a power cord for the device away from traffic.

big_patent

en

Summarize: The Oregon Court of Appeals ruled Wednesday that a southern Oregon couple must quiet their incessantly barking dogs by sending them to the vet to have their voices surgically squelched. The Appeals Court ruled “debarking” surgery is an appropriate solution to a noisy and relentless problem that neighbors living next to the dogs have had to endure for more than a decade on their rural property outside Grants Pass. Debarking operations, also known as devocalization, are highly controversial. Groups such as the Oregon Humane Society and American Humane have spoken out against them. Six states have outlawed the procedure under certain circumstances, according to the American Veterinary Medical Association. The surgery involves cutting the vocal cords. Opponents say removing a dog or cat's prime means of communication is cruel and unnecessary. Proponents say if done correctly, it can save problematic animals from being euthanized and still allow them to express themselves with a soft, raspy bark or muffled squeak. But it’s rare for courts in Oregon to order the procedure done, in part because barking-dog disputes usually are resolved long before cases get that far. “We are just shocked,” said David Lytle, a spokesman for the Oregon Humane Society. Lytle said his organization pushed for a bill to outlaw debarking surgeries in Oregon, but it failed a few years ago. The lawsuit began as a last resort, according to the neighbors who filed it. Debra and Dale Krein said they could no longer take the barking of the six or more Tibetan and Pyrenean Mastiffs owned by the couple who lived next door for almost 20 years. The barking started in 2002, but the Kreins didn't sue Karen Szewc and John Updegraff until 10 years later, according to a court summary of the case. Like the Kreins, Szwec and Updegraff are married. The Kreins contended the barking started as early as 5 a.m. and continued for hours on end after Szewc and Updegraff left the house for the day. The dogs routinely roused the Kreins from sleep, deterred relatives from visiting their property and forced them to turn up the volume of their TV to watch shows, they said. Their children dreaded coming home from school. The Kreins made audio recordings to prove their case. After a four-day trial in Jackson County Circuit Court in April 2015, a jury ruled that Szewc and Updegraff had to pay the Kreins $238,000. The Kreins at the time also argued that while the money compensated them for several years of disruption, it didn’t stop the problem. Judge Timothy Gerking agreed and ordered that the Mastiffs be debarked, given that the owners hadn't stopped the barking by other means, including using citronella-spray and shock collars or erecting a visual barrier between the dogs and the neighbors’ property. The Appeals Court upheld the $238,000 verdict and Gerking’s ruling, reasoning that the Kreins shouldn't have to file lawsuit after lawsuit to recover compensation as the problem continues. In his written opinion, Appeals Judge Joel DeVore likened that to a “judicial merry-go-round.” Reached by phone, Debra Krein declined comment. The Kreins' Medford attorney, Michael Franell, couldn't be reached for comment. Szewc told The Oregonian/OregonLive that efforts to silence her dogs have threatened her ability to run her farm. “The dogs are my employees,” she said. “We do not have the dogs to harass the neighbors. We have the dogs to protect our sheep.” The dogs bark, she said, when they sense predators, such as bears and cougars. She said agricultural properties generate farm noise -- something her neighbors haven't come to accept. “The next line of defense is a gun. I don’t need to use a gun, if I can protect my sheep with dogs," Szewc said. "This is a passive way of protecting livestock.” Szewc said she and her husband currently have six dogs, but the number has fluctuated over the years. In 2005, Jackson County cited Szewc for allowing two of her dogs to become a public nuisance with frequent and prolonged barking. After reviewing the case, county hearings officer Donald Rubenstein in 2006 ordered Szewc to pay a $400 fine and have the dogs debarked or moved off of her land. In making his decision, Rubenstein found that Szewc’s farm activities then were so small and unprofitable that they didn't fall under farm-use laws that might have protected the sound of the barking dogs. Szewc and Updegraff have strongly disputed that. Szewc said the farm made $26,000 last year and it has supplemented their income. Court papers describe the couple's land as a 3.4-acre parcel, populated by sheep, goats and chickens. Szewc said the couple did debark the dogs, but it had disastrous effects in 2010. A cougar ran off with six lambs in a single week, she said. “That’s $3,000 of income,” Szewc said. Now, Szewc said, she doesn’t know what she’ll do -- whether she’ll try to appeal the decision or accept it. Only one of the couple's six dogs have been debarked. The ruling was made by a three-judge panel of the Appeals Court: Joel DeVore, Chris Garrett and Bronson James. Read the opinion here. -- Aimee Green agreen@oregonian.com o_aimee Full Width Column 1 Decisions of the Oregon Supreme Court, Court of Appeals, and Tax Court are posted weekly after 8:00 a.m., or as soon as available, on the day issued by the court. Final publication in the bound volumes of the Oregon Reports are accessible through the State of Oregon Law Library Digital Collection. Opinions Owners must surgically 'debark' loud dogs, court rules https://t.co/cnggmgZZdj pic.twitter.com/148pp9Y9mk — The Oregonian (@Oregonian) August 31, 2017 An Oregon appeals court agreed Wednesday that a couple must surgically lacerate their dogs’ vocal cords in a procedure known as “debarking” or “devocalization,” following a lawsuit brought by neighbors annoyed by the pets’ “incessant barking.” The ruling upheld a lower court order. The case began in 2002, when Karen Szewc and John Updegraff began breeding Tibetan Mastiffs, large fluffy dogs often employed to protect sheep from predators, at their home in Rogue River, Ore., about 150 miles south of Eugene. The married couple’s neighbors, Debra and Dale Krein, quickly grew tired of the dogs’ barking. According to the Kreins, the “dogs bark[ed] uncontrollably for long periods of time while defendants [were] away from the residence,” court documents state. But they weren’t the first ones to take action against the dog owners. In both 2004 and 2005, Jackson County cited Szewc for violating a county code provision on public nuisance “by allowing two of her dogs to bark frequently and at length,” according to court documents. Szewc argued the provisions didn’t apply to her because she ran a farm on the couple’s 3.4-acre parcel of land, which includes sheep, goats and chickens. Farms fall under different ordinances. The Jackson County Circuit Court rejected this argument, saying the property was not a farm, ordered her to pay $400 and to debark the two offending dogs or to move them to a different area. It is unclear if she debarked these dogs, but in 2012, the Kriens filed a lawsuit against Szewc and Updegraff, claiming they had not taken the necessary actions to prevent the dogs from barking. At that point, there were at least six dogs on the property, all either Tibetan or Pyrenean Mastiffs, the Oregonian reported. Again, the dog owners argued that they were not subject to the dog barking ordinance because they were running a farm. The Kreins claimed the dogs often began barking at 5 a.m., sometimes waking the couple. Relatives refused to visit, and their children hated being around the house, according to the Oregonian. They recorded the barking to prove it. “The dogs are my employees,” Szewc told the Oregonian. “We do not have the dogs to harass the neighbors. We have the dogs to protect our sheep.” “The next line of defense is a gun. I don’t need to use a gun, if I can protect my sheep with dogs,” she added. “This is a passive way of protecting livestock.” In April 2015, a jury sided with the Kreins and ordered Szewc and Updegraff to pay them $238,000 in damages. Also in response to the suit, Judge Timothy Gerking ordered the couple to debark the mastiffs, since they hadn’t stopped them from barking using other means such as shock collars. Szewc and Updegraff again argued unsuccessfully that the dogs were necessary because they had a farm. On Wednesday, a three-judge panel of the Oregon Court of Appeals consisting of Joel DeVore, Chris Garrett and Bronson James upheld that ruling, agreeing that the dog owners were not running a farm. The question of whether debarking is an appropriate remedy was not at issue in the case. Debarking is a surgical procedure in which parts of a dog’s vocal folds or cords are cut out in an effort to lower the volume of its barks or, more severely, to eliminate the dog’s ability to bark altogether, according to the American Veterinary Medical Foundation. The procedure is partially prohibited in six states, according to the AVMF. Many animal welfare organizations oppose it, as do some veterinarians. “Debarking is not a medically necessary procedure,” Jeffrey S. Klausner, chief medical officer of the Banfield Pet Hospital, told the New York Times in 2010. “We think it’s not humane to the dogs to put them through the surgery and the pain. We just do not think that it should be performed.” Wednesday’s ruling left some animal rights activists reeling. “We are just shocked,” David Lytle, a spokesman for the Oregon Humane Society, told the Oregonian. More from Morning Mix Trump and Manafort get big reminder that pardon power does not extend to state crimes Investors say SeaWorld lied about business downturn after orca outcry. Now feds are investigating. Evangelicals’ ‘Nashville Statement’ denouncing same-sex marriage is rebuked by city’s mayor Lesbians win $10,000 judgment against county clerk for calling them an ‘abomination’ MEDFORD-- An Oregon couple who sued their neighbors because of their constantly barking dogs was awarded nearly $240,000 in damages. The Mail Tribune reports that a Jackson County jury found in favor of Dale and Debra Krein of Rogue River, awarding damages for what was described as more than a decade of ceaseless barking. The defendants, John Updegraff and Karen Szewc, said their dogs were necessary to protect their livestock from predators. The court last week found that the Tibetan mastiffs weren't ideally suited to be livestock guardians and ordered them debarked within 60 days or replaced with a more suitable breed. Szewc and Updegraff reportedly began breeding the dogs at their home around 2002. According to the Mail Tribune, the Kreins claimed the dogs would often begin barking at 5 a.m. and would continue throughout the day. They also claimed the couple did nothing to quiet their dogs even after being cited by Jackson County Animal Control in 2002 and 2004 for violating public nuisance codes. -- The Associated Press

Summary: "We are just shocked." That's the reaction of the Oregon Humane Society to a Wednesday ruling by the Oregon Court of Appeals that will require a Rogue River couple to have their six dogs' vocal cords cut. The court case was one between neighbors of nearly 20 years, reports the Oregonian: Debra and Dale Krein said the barking from Karen Szewc and John Updegraff's Tibetan and Pyrenean Mastiffs began in 2002-at 5am each day, and didn't stop. The suit was filed 10 years later, with the Kreins alleging an auditory hell so bad their kids didn't want to come home from school. Szewc and Updegraff reportedly did attempt to rectify the situation with methods including shock collars, to no avail. A jury ruled in the Kreins' favor in 2015; Szewc and Updegraff were ordered to pay the couple $238,000 and have their dogs undergo devocalization. The Appeals Court upheld that ruling. Szewc and Updegraff had argued that the dogs were what kept their livestock-sheep, goats, and chickens on 3.4 acres-safe from predators. The Washington Post reports that as such, they argued the county's public nuisance code didn't apply as they were subject instead to farming ordinances. But the AP in 2015 reported on the original ruling, which found Tibetan mastiffs aren't a breed designed to guard livestock. It also cited the 2006 decision of a local hearings officer (related to a citation over the barking) who found the farm use defense was unavailable to the couple due to the small size and profits of their farming endeavors. They have not decided whether to appeal the latest decision. The Oregonian notes the surgery still allows dogs to make a very quiet bark or squeak; critics call it a "cruel and unnecessary" procedure.

multi_news

en

Summarize: A former homeless crack addict who found fame four years ago after a YouTube video catapulted him to overnight fame has revealed that he is broke despite a six-figure book deal and a voice over contract with Kraft. Ted ‘Gold Voice’ Williams became a media sensation when he was filmed panhandling in Columbus, Ohio in December 2010 with a sign reading: 'I'm an ex-radio announcer who has fallen on hard times'. The once-successful DJ was recorded performing a voice-over by a local reporter and the video went viral, receiving more than 20 million views on YouTube. Scroll down for video. Ted ‘Gold Voice’ Williams, a former homeless crack addict who found fame four years ago after a YouTube video of him went viral, has said he is broke despite a $375,000 book deal in 2012. With his story garnering national coverage, Williams found himself with a home, radio job offers - and money for the first time since he had moved into a tent in the woods 17 year earlier as an addiction to crack cocaine took over his life. The notoriety that followed his amazing comeback led to numerous voice-over jobs, including one as the spokesman for Kraft Macaroni and Cheese. Two stints in rehab have helped him deal with first his cocaine abuse and then issues with alcohol, but problems remain in his life. 'I should have been a millionaire by now,' he told The Columbus Dispatch after a recent appearance on a Marion, Ohio, radio station, where he has a weekly show. Williams became a media sensation when he was filmed panhandling in Columbus, Ohio in December 2010 with a sign reading: 'I'm an ex-radio announcer who has fallen on hard times' Inspiration: Williams now spends part of his time visiting homeless shelters and offering support to other people in the position that he once found himself. Despite receiving a $375,000 book advance for a biography, A Golden Voice: How Faith, Hard Work, And Humility Brought Me From The Streets To Salvation, Williams admits he is broke. 'I own nothing,' he said, adding that he doesn't have furniture or a car. The situation was very different in 2011 when Williams first found fame and lived in hotels paid for by an agent. After that he moved into a rented condominium owned by another agent, but now he lives in an apartment and admits he owns no furniture. ‘Financially I'm a little under the weather. I lot of things I signed in 2011 I probably shouldn't have signed,’ he said. Williams admits he ‘wasn't focused’ and ‘trusted too much’ in managers who he now believes didn't have his best interests at heart. He now says his Christian faith has helped him cope with his resentment and anger about what has happened. 'I’m still very fragile in my recovery,' he said. In the 1980s, Williams had been a successful radio show host on a top-rated morning show in Columbus, but then he slid into a life of homelessness and drug abuse for 17 years and lived in a tent. In the 1980s, Williams had been a successful radio show host on a top-rated morning show in Columbus, but then he slid into a life of homelessness and drug abuse for 17 years until the YouTube video gave him a second chance. Williams has had other struggles since 2010 and initially turned to alcohol. 'I figured since it wasn’t my drug of choice, alcohol could be my new drug,' Williams told the Today show in 2012. 'I could go and start drinking, and nobody would know. Everybody would know [if] Ted was on crack, but they wouldn’t know that Ted was drinking.' When he appeared on the Today show he had just achieved one year of sobriety and was celebrating by walking his daughter down the aisle at her wedding. 'I was able to be a part of that, something that a year and a half ago I wouldn’t have even thought about, let alone become a part of,' he said. Mug shots: Williams was also arrested several times for charges including robbery and possession of drugs

Summary: Ted 'Gold Voice' Williams, a former homeless crack addict found fame four years ago after a YouTube video of him went viral. Catapulted to overnight fame, he received a $375,000 book deal and continues to do voice over work for Kraft. But Williams says that bad management mean he is now broke and doesn't own either furniture or a car. 'Financially I'm a little under the weather. I lot of things I signed in 2011 I probably shouldn't have signed,' he said. In the 1980s Williams had been a successful radio show host before he slid into a life of homelessness and drug abuse for 17 years. He says his Christian faith has helped him cope with his resentment and anger about what has happened. Since finding fame in 2011 he has been in rehab twice - first to overcome his addiction to crack cocaine and then to over an issue with alcohol.

cnn_dailymail

en

Summarize: FIELD OF THE INVENTION [0001] This invention relates to steel shelving and more particularly to a steel shelving unit providing both adjustable shelving as well as accessory functions for holding items other than on a shelf. BACKGROUND OF THE INVENTION [0002] Steel shelving units comprising horizontal shelf-supporting beams with ends adjustably connected to vertical support columns for shelf height adjustment are known. While such units are very useful, it is desirable to provide such beams with improved structures for converting such beams to such columns. As well, it is frequently desirable to provide additional capacity in such units for holding items other than by positioning on the shelves themselves. Additionally, it is desired to provide for vertical adjustability of shelf unit accessories providing such additional capacity. SUMMARY OF THE INVENTION [0003] To these ends, a preferred embodiment of the invention includes an improved support column for a shelving unit with provisions for adjustably mounting shelf-supporting beams and accessories, such as a removable, adjustable, hook accessory for hanging items for storage or display, other than on the shelves of the unit. [0004] Accordingly, an improved support column according to a preferred embodiment of the invention includes a plurality of perforations or apertures in the column face and which operably accommodate end brackets of a horizontal shelf-supporting beam as well as removable accessories, such as hooks which cooperate with the apertures. [0005] Each perforation in the column is defined by a plurality of edges which define tapered support surfaces for cooperating with beam end brackets to adjustably mount the brackets in position up and down the columns. Transverse edges of the perforations or apertures are preferably bent or curved to support a bracket component of an accessory such as a hook. Both the beam-end bracket and the hook bracket interface with two adjacent, vertically-spaced, apertures in the column for securely holding them in a selected vertical position on the column. [0006] Preferably the perforations are symmetrical on the column face, and centrally disposed therein so a column can be used on both right and left sides of the shelving unit with all columns in the unit (typically four of them) preferably identical. [0007] The beam-end brackets are each angular or L-shaped, with one leg welded or affixed to the beam, and the other leg for lying adjacent to or on the column face over at least part of the column aperture. This other bracket leg preferably has tabs for extending into and engaging a tapered edge of each of two column apertures, one above the other, to hold the bracket and its fixed beam in a fixed, but removable position at selected vertical positions on the column. [0008] The hook bracket has extensions, one fitting over a curved portion of an upper aperture on the column and another over a similar curved portion of a next lower aperture for securing the hook in the column face at a selected vertical position. A tab, upwardly extending from the hook bracket, engages a rear side of the column face at the upper aperture to facilitate mounting ad holding the hook and accessory thereon. [0009] Other accessories, similarly mountable to the columns, and other than hooks, are contemplated. [0010] Accordingly, the invention provides a shelving unit having vertically adjustable shelf-supporting beams, as well as vertically adjustable accessories, both mountable on a supporting column at variable, selected, vertical positions. Improved perforation on the faces of the columns facilitates both beam and accessory mounting. [0011] These and other objectives and advantages will become readily apparent from the following detailed description of a preferred embodiment of the invention and from the drawings in which: BRIEF DESCRIPTION OF THE DRAWINGS [0012] FIG. 1 is a front isometric view of the overall shelving unit of the invention; [0013] FIG. 2 is a front isometric view of a column of the invention; [0014] FIG. 3 is a front isometric view of a horizontal shelf-supporting beam of the invention; [0015] FIG. 4 is a fragmentary isometric cross-sectional view generally taken along lines 4 - 4 of FIG. 3 ; [0016] FIG. 5 is a rear isometric view of the fragmentary isometric cross-sectional view of FIG. 4 ; [0017] FIG. 6 is an enlarged isometric view of the encircled area of FIG. 1 ; [0018] FIG. 7 is a front partial view of the beam and post of FIG. 6 ; [0019] FIG. 8 is a rear view of the partial beam and post of FIG. 7 ; [0020] FIG. 9 is a cross-sectional view taken along lines 9 - 9 of FIG. 8 ; [0021] FIG. 10 is a front isometric view of a hook accessory spaced from a column shown in partial view; [0022] FIG. 11 is an isometric view as in FIG. 10, but showing the hook accessory mounted to the column; [0023] FIG. 12 is a cross-sectional view taken along lines 12 - 12 of FIG. 11 but showing the respectively rotated hook accessory being partially mounted to a column; and [0024] FIG. 13 is a view similar to FIG. 12 but showing the hook accessory mounted to the column. DETAILED DESCRIPTION OF THE INVENTION [0025] Turning now to the drawings, there is shown in FIG. 1 an isometric view of a dual function shelf unit 10 according to one embodiment of the invention. Shelf unit 10 includes four vertical support columns 11 - 14, a plurality of horizontal shelf-supporting beams 15 - 22, a plurality of shelves 23 - 26 which are shown as wire-formed shelves but could be in other wire patterns, of solid material, of grate or other patterns or of any suitable shelf configuration, each supported by one of beams 15 - 18 and one of beams 19 - 22, respectively. [0026] A plurality of horizontal braces 27 - 30 extend between respective columns 12, 13 and 11 - 14 as shown, as well as a plurality of slanted braces 31, 32 as shown. [0027] Optionally, a plurality of preferably identical tie bars 33, 34, 35 extend between respective ones of the front beams 15 - 18 and rear beams 19 - 22 to support shelves 23 - 26 respectively, these bars 33 - 35 extending respectively under each shelf and supported on the respective horizontal beams, particularly on flange 78 (see FIGS. 4-6 ). [0028] A column 12 is illustrated in FIG. 2, column 12 being preferably identical to columns 11, 13, 14 and thus only one being described in detail, each with preferably like parts. Identical construction allows a column to be used at any corner position in unit 10, [0029] Columns 11 - 14 are in a C-shaped cross-sectional configuration (see also FIGS. 6, 7, 8, 10 - 13 ). Each column thus has a face 40, a plurality of symmetrical apertures 42 in the face 40, and a plurality of rectangularly-shaped apertures 44 in each leg 46, 48. Each leg terminates along its legs 46 - 48 with its flanges 50, 52 completing configuration of column 12 (see FIGS. 2, 6 and 9, for example). [0030] Details of apertures 42 are perhaps best seen in FIGS. 6-13. Referring first to FIG. 6, each aperture 42 is preferably identical to each other aperture 42 in the respective columns. Each aperture 42 is oriented one above the other in the columns, equally spaced vertically and aligned centrally and symmetrically in column face 40, as shown. Each aperture is defined in part by edges 54, 56, tapering or inclined from a bottom edge 58, 60 upwardly and outwardly in face 40 to upper edges 62, 64. [0031] A curved tab 66 extends upwardly between bottom edges 58, 60 and is turned or curved inwardly from face 40. A second tab 68 extends downwardly as part of face 40 between edges 54, 56 and has an inwardly curved tab end 70, as shown. [0032] Tapering or inclined edges 54, 56 may be continuously straight, or may define slightly different angles, such as at breaks 72, 74 as illustrated in FIG. 6 such that upper regions of edges 54, 56 incline outwardly in face 40 at a slightly greater angle than lower regions of those edges to facilitate initial bracket engagement. [0033] Horizontal beams 15 - 22 are, like columns 11 - 14, each preferably identical to each other so they can be interchangeably used in shelf unit 10. Only one exemplary beam 17 will be described in detail, [0034] With initial reference to FIGS. 3-5, an elongated shelf support beam 17 includes a preferably embossed face 76, having a plurality of outstanding ornamental projections 77 of any suitable form embossed, pressed from or otherwise defined in face 76. [0035] Beam 17 is defined by a lower, rearwardly extending flange 78, a first rearward extending upper flange 79, a return 80 and a final rearwardly extending shelf-supporting flange 81. Other beam configurations may be used but this beam provides both shelf-supporting flange 81 and lower, tie-bar supporting flange 78 as will be described. [0036] To an end of beam 17 is welded (or otherwise fixed) an L-shaped bracket 82 having a rearwardly extending leg 83 and a front leg 84. A bracket 82 a, which is essentially a mirror image and otherwise identical to bracket 82, is fixed to the other end of beam 17 and is otherwise similar to the bracket 82. Similar parts of the respective brackets are numbered with a suffix “a”. Legs or faces 84, 84 a of brackets 82, 82 a are provided with punched-out upper and lower locking tabs 87, 88, 87 a, 88 a formed at an angle in leg 84, 84 a, respectively ( FIGS. 4, 5 ). As will be described, these tabs serve to engage and interface with inclined edges 54 of adjacent apertures 42 (see FIGS. 8, 9 ) holding and locking the brackets 82 to column 12. Similar tabs 87 a, 88 a will engage and interfere in similar fashion with inclined edge 56 of apertures 42 in another column 11 at the other end of the beam to hold and lock the bracket 82 a and beam 17 to that column 11. [0037] FIGS. 6-8 further illustrate the interconnected relationship of a beam 17 to a column 12. The other beam end is supported and locked such as on a column 11 by a bracket 82 a operational in a similar fashion. [0038] In FIG. 6, bracket 82, via tabs 87, 88 is locked onto column 12. For example, the beam 17 and its fixed bracket are oriented proximate vertically adjacent apertures 42 in column 12, with tabs 87, 88 initially proximate upper ends of apertures 42 so the tabs can be inserted into the apertures. When leg 82 is against face 40, the bracket (and beam) is moved downwardly, tabs 87, 88 engaging and fitting around inclined edges 54 in the respective apertures. The distance between tabs 87, 88 and the leg 82 of the bracket causes, as the bracket moves downwardly, the tabs 87, 88 and leg 84 to be frictionally wedged onto column 12, the engagement of respective tabs 87, 88 with edges 54 providing frictional, locking engagement of bracket 82 and beam 17 to column 12. Similar complementary action of tabs 87 a and 88 a with inclined edges 56 of vertically-adjacent apertures 42 in column 11 secures the other end of the beam in vertically-coordinated position so the beam 17 is horizontally supported across and between columns 11 and 12 as described. [0039] The vertical locations of the beams 15 - 22 can be set on the columns as desired to provide the eventually desired spacing between any shelves as described in unit 10. [0040] It will be further appreciated that the frictional interface between two respective apertures 42 in each column, and the single complimentary brackets at each end of the beam strengthens and rigidifies any tendency of the columns connected by the beams to “rack”, move or tilt toward or away from one another, resulting in a very strong, rigid unit 10 construction. This benefit is, in part, also provided by the engagement of the inner faces of legs 83, 84 with the respective leg 46 and face 40 of column 12 as well as tabs 87, 88 and inclined aperture edge 54. Complementary engagement of complementary parts of bracket and column at the other beam end provide the same result. [0041] Accordingly, it will be appreciated that a shelf unit 10, as described above, provides a rigid shelving function for a variety of applications. [0042] In a further embodiment of the invention, a further support function is provided by the addition of an accessory which provides a further article support or hanging function shelving unit 10. [0043] This additional embodiment is illustrated in FIGS. 10-13 of the drawings, wherein an accessory hook 90 is provided for use on a column, such as a column 12 as described above. In FIG. 10, hook accessory 90 includes a hook member 91 preferably rigidly-mounted at shank 91 a to a hook bracket 92. Bracket 92 includes a face surface 93, a lower flange 94 with a depending tab 95, and upper flange 96 with a downwardly depending tab 97, and an upwardly extending locking tab 98. [0044] FIG. 10 illustrates a column 12 and a hook accessory 90, not yet assembled and FIG. 11 illustrates a hook accessory 90 attached to a column 12. In FIG. 11, it is noted that flanges 94 and 96 of hook accessory 90 rest on the curved tabs 66 respectively of vertically-oriented adjacent apertures 42 in face 40 of column 12. Tabs 97, 95 prevent hook accessory 90 from being pulled outwardly, away from column 12. Locking tab 98 prevents removal of hook accessory 90 horizontally and forwardly of column 12 since it engages or is in close blocking proximity to curved end 70 of tab 68 if moved directly forwardly. [0045] Referring to FIGS. 12 and 13, FIG. 12 illustrates the preferred motion for attaching accessory hook 90 to a column such as 12. In use, the upper end of hook 90 is inserted into an upper aperture 42, tab 98 extending therein below, then behind end 70 of tab 68 in the aperture. Once the upper end of accessory hook 90 is extended into upper aperture 42, the lower end of bracket 92 can be rotated into a lower aperture 42 and then bracket 92 lowered so tabs 97, 95 are locked behind the curved tab ends 66, respectively of the vertically-adjacent apertures as in the cross-sectional view of FIG. 13. [0046] Hook accessory 90 is thus removably but securely mounted at selected vertical positions up and down column 11, 12 and others, providing for additionally supporting functions for a variety of items on shelving unit 10. In this embodiment, the hooks can be selectively spaced along the entire lengths of the columns excepting at the same position of the brackets 82, 82 a on the columns for the horizontal shelf-supporting beams. [0047] Vertically-adjustable storage is thus not limited to the shelves only but includes the additional function of hanging items suitably on vertically-adjustable hooks, [0048] It will be appreciated that other storage or hanging accessories can be similarly attached to the columns to provide additional hanging or storage functions,

Summary: A dual function shelf unit has identical support columns, horizontal shelf-supporting beams adjustably-mounted on the columns for vertical shelf adjustment and one or more hooks also mountable on at least one column. The columns have apertures with complementary apertures having opposed inclined edges for accepting a beam-end bracket from either side of the column and tabs for accepting mounting flanges of hooks. Both beam-end brackets and hooks are each mounted in two vertically-spaced apertures. Vertically adjustable shelf and hook storage is provided.

big_patent

en

Summarize: BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates to a sink spray. More particularly, it refers to a sink spray capable of engaging multiple attachment devices to vary the utility of the sink spray. 2. Description of the Prior Art Sink sprays are well known products and are currently found as a fixture in most private houses, apartments and commercial kitchen facilities. Typical sink sprays are described in U.S. Pat. Nos. 3,498,546; 4,148,438; and 4,344,578. Such sprays are adapted for one use; namely, portability for directed discharge of water at the end of a limited length hose. Because space is restricted on a sink and plumbing codes frequently limit fixtures on a sink, the single purpose use of a sink spray is inconvenient. When a below sink filter system is employed it could be hooked up to the sink spray but this eliminates the spray from use as a utility device to clean pots and pans. Moreover, the spray nozzole on the sink spray is not a convenient device for filling a glass or other drinking container. A way is needed to easily convert a sink spray into a multitude of uses. SUMMARY OF THE INVENTION I have invented a sink spray that can be easily mated to various utilitarian devices useful on a sink. My apparatus mates a sink spray device with a spigot for use when the water circulation system feeds through a below the sink water filter device. Alternatively, a brush can be mated to the sink spray to provide means for cleaning pots, bottles and pans using both the abrasive power of the brush and the concentrated water pressure from the spray device. The sink spray unit contains an annular hose connector attached to a sink&#39;s standard flexible conduit. The annular hose connector is in turn attached to a three component housing containing a two-way valve and means for rotatably attaching the valve housing to the attachments. The attachment can be a spigot which causes water to flow when a locking tab in an annular opening to the spigot engages a slot in the sink spray housing and actuates a valve to permit water flow. Alternatively, a brush attachment to the sink spray with the locking tab in its annular opening merely acts to seat the brush on the sink spray. The handle on the sink spray is used to actuate the valve in the sink spray housing to allow water to flow through the brush attachment. The sink spray consists of three integral parts; namely, an upstream housing, a valve housing and a downstream housing. The upstream housing is connected to a flexible hose from a water source, the valve housing contains the two way valve controlling water flow and the slot for receiving the locking tab from the auxiliary attachments. The downstream housing mates with the interior of each attachment device. BRIEF DESCRIPTION OF THE DRAWINGS The present invention may be best understood by those having ordinary skill in the art by reference to the following detailed description when considered in conjunction with the accompanying drawings in which: FIG. 1 is a perspective view of the sink spray and its attachments in relation to a sink. FIG. 2 is an elevation view in partial section of the sink spray and spigot attachment in an operational mode. FIG. 3 is an elevation view in partial section of the sink spray in a non-operating mode. FIG. 4 is a plan view of the sink spray. FIG. 5 is a side elevation view partially in section showing the sink spray and spigot attachment in a non-operating mode. FIG. 6 is a side elevation view partially in section showing the sink spray and spigot attachment in an operating mode. FIG. 7 is a detailed sectional view of the sink spray two way valve closed and the valve housing exterior surface features when the apparatus is in the FIG. 5 non-operating mode. FIG. 8 is a detailed sectional view of the sink spray two way valve open and the valve housing exterior surface features when the apparatus is operating according to FIG. 6. FIG. 9 is a diagramatic view of the water flow system going through or by-passing the filter element below the sink. FIG. IO is a diagramatic view of the water flow system going through a filter element below the sink. FIG. 11 is an elevational view in section of an alternate sink spray from the one shown in FIG. 3. FIG. 12 is a detailed elevation view partially in section of the annular hose connector. DETAILED DESCRIPTION OF THE INVENTION Throughout the following detailed description the same reference numerals refer to the same elements in all figures. The apparatus of this invention combines a sink spray 10 with at least two alternate attachments 12 or l2A. The sink spray 10 is generally cylindrical in shape and can be conveniently made from injection molded high density polymer materials such as DuPont DELRIN® or Celanese CELCON®. Three basic parts of the spray housing include the valve housing 14, an integral cylindrical upstream stem 16 and an integral cylindrical downstream stem 18. The valve housing 14 has an actuating lever 20 (FIGS. 2-3) inserted through an opening 22 to the interior of valve housing 14 where it is capable of actuating a valve 24. Alternatively, the actuating lever can be a grip 20A with groove 104 (FIG. 11) engaging the top 54 of valve 24. The valve housing 14 has a lock tab slot 26 which accommodates a locking tab 28 or 28A from the inlet end 30 or 30A of the attachment 12 or 12A. Movement of the locking tab 28 or 28A through entrance groove 102, past notch 103 (FIG. 4) places it in a first position 32. When tab 28 or 28A is moved to a second position 34 it opens valve 24. See FIGS. 7 and 8. The attachment 12 has an inverted J-tube 70 leading from a cylindrical body 74 integral with an annular member 72. Annular member 72 receives horizontal stem 18 from sink spray 10. Water flowing into attachment 12 exits at opening 76. Attachment l2A has cylindrical body 74A integral with annular member 72A. Annular member 72A receives horizontal stem 18 from sink spray 10. Brushes 78 are integral with body 74A. The vertical stem 16 of the sink spray 10 is threaded at its upstream end to an annular hose connector 36. The annular hose connector 36 separates the vertical stem 16 from the flexible water conduit 42 which in turn is attached to a water supply. This conduit 42 provides the means for the water supply to reach the sink spray 10. The standard features of the hose parts including flexible hose 42, metal hose connector 40 and its nylon bushing 90 are connected over the downstream end of connector 36 by insertion of a metal snap ring 84. A washer 82 downstream from ring 84 prevents leakage at this connection. The downstream end of connector 36 has male threads 41 which engage the upstream threads on the inner surface of vertical stem 16. For purposes of this specification the water source in the sink piping 68 is the furthest upstream point and the water outlet 76 is the furthest downstream point. The flexible water conduit 42 moves up and down through receptacle 44 attached to sink 46. Its lowest position is determined by the upstream edge 38 of the vertical stem 16 which rests on the top surface 45 of the receptacle 44. The flexible water conduit 42 can be connected to any water supply source containing potable water. Valve 24 is normally in the closed position because of the action of spring 50 held in place by retainer 83. Seal 80 prevents leakage of water. See FIG. 3. Movement of lever 20 in a direction towards the horizontal stem 18 causes an actuating member 52 attached to the lever 20 to press down on the downstream nipple 54 of valve 24. In like manner, squeezing of handle 20A causes pressure to be exerted on nipple 54. This causes the valve 24 to be depressed against its spring 50 and opens chamber 56 to allow the flow of water from hose 42. See FIGS. 2 and 9. When the sink spray 10 is disconnected from an accessory device 12 or l2A, actuation of lever 20 or handle 20A causes water to spray from nozzle end 60. When in the upright position, lever 20, as shown in FIG. 3, has no action on valve 24 and consequently no water flows through the sink spray 10. Upon insertion of the horizontal stem 18 into inlet 30 or 30A and fully seating it by inserting tab 28 or 28A into entrance groove 102, the system is ready for operation. The mechanism for allowing the flow of water to move from conduit 42 when the accessory 12 or l2A is in place is not lever 20 or 20A but locking tab 28 or 28A which moves from a first position 32 as shown in FIG. 7 to a second position 34 as shown in FIG. 8. In position 34 the tab 28 depresses the actuating member 52 and causes valve 24 to be pressed against its spring 50 and thereby opens channel 56 to allow the flow of water. FIG. 5 shows the sink spray 10 and spigot 12 in a resting position and FIG. 6 shows the spigot 12 pulled down about 30 degrees to actuate valve 24 in the sink spray 10 and permit water to flow through channels 56 and 57 through porous material 106, and channel 68 to outlet 76. The large filter container 66 as shown in FIGS. 9 and 10 is designed to be installed below the sink to provide filtered water through spigot 12. A switch 98 on sink 46 changes the flow through valve 64 so that the water flows through piping 62, through filter 66 and eventually out through outlet 76. If the sink spray alone is to be used without the filtering cycle, the switch 98 is turned so that water flows through piping 108 directly to the sink spray. See FIG. 9. A simple piping diagram with water flow from its source piping 68 through piping 62A to filter 66 is shown in FIG. 10. Lock ring 84 holds the hose connection in place downstream from the annular hose connector 36 as seen in FIG. 12. Nylon bushing 90 contains a groove for holding the lock ring 84. A standard hose fitting 40 is connected to nylon bushing 90 to make the water connection. O-rings 96 in the horizontal stem 18 prevent leakage of water. The auxiliary attachment devices can be molded from the same or different polymer materials used to make the sink spray housing. The filter cartridge can contain activated charcoal, coral sand, ion exchange resins or other filter media. The water filter container 66 can also contain additive ingredients downstream of the filter such as calcium or magnesium to add desirable minerals to the water. Various flavorings can be added to the water outlet 76 so that the water runs through the flavoring and gives added taste to the water. Equivalent valves and mechanisms can be substituted for the various valves and mechanisms of the present invention without departing from its scope.

Summary: A multipurpose sink spray device having an upstream and downstream stem integrally joined to a valve housing containing a two-way valve. A slot in the valve housing receives a locking tab mounted in an annular opening of an auxiliary attachment device such as a spigot or a brush. Movement of the locking tab in the slot activates a valve in the housing and allows water to flow through the attachment device. Alternatively, the sink spray device can be activated without the auxiliary attachment device by depressing a lever or handle on the sink spray housing.

big_patent

en

Summarize: Two San Diego teenagers were killed execution-style in a triple homicide Sunday morning at a Tijuana apartment complex, and Mexican authorities were continuing to search for the assailants. The victims were Christopher Alexis Gomez, 17, a football player in his senior year at O’Farrell Charter High School, and Juan Suarez-Ojeda, 18, who graduated from Ingenuity Charter School on the shared Encanto campus earlier this year. The third victim was Angel Said Robles, an 17-year-old Tijuana high-schooler. Gomez’s cousin, Katheryn Garcia, said the trio had gone together to a barbecue in Ensenada last Friday and were supposed to return that same night. A Tijuana police detective told Gomez’s family the three teens were tortured before they were shot Sunday, Garcia said. There was no initial indication of what might have led to the brutal killings. Courtesy of O'Farrell Charter School Staff at O'Farrell Charter School set up a GoFundMe for the families of Juan Suarez-Ojeda, left, and Christopher Gomez, right. Staff at O'Farrell Charter School set up a GoFundMe for the families of Juan Suarez-Ojeda, left, and Christopher Gomez, right. (Courtesy of O'Farrell Charter School) Family photo Angel Said Robles Angel Said Robles (Family photo) The incident happened in Lomas Verdes, a high-crime area in central Tijuana with high rates of homicide, neighborhood drug dealing and domestic violence. The semi-clothed bodies of the three victims were found early in the morning in a complex of apartments. The bodies were found outside one of the buildings, and initial police reports stated they had been shot in the head. Mexican authorities said the San Diegans had no criminal record in Tijuana, but Said had a car theft on his record — a statement Said’s family called “a lie.” Jorge Alvarez, head of the Baja California Attorney General's Office in Tijuana, withheld some details in an interview Thursday, citing the ongoing investigation. But he said that preliminary information indicated that the two San Diegans were familiar with the neighborhood where they were killed. “They did not live there, but they came to visit family members, one of them apparently did so frequently,” Alvarez said. Said’s uncle, Juan Carlos Contreras of San Diego, said his nephew lived in the neighborhood, as did Suarez-Ojeda’s grandmother. The boys had been friends for several years, he said. As for Gomez, Katheryn Garcia contradicted law enforcement and said her cousin had never been to Tijuana or Ensenada before this trip. Gomez was born and raised in San Diego and was not well-traveled, his cousin said, adding that he’d been to Mexico only once before, when their families traveled together in 2016 to visit other family members in Morelia, the capital of the central state of Michoacan. Gomez’s cousin described the teen as “an innocent boy” who was the “sweetest, most selfless person.” He recently got his first job cleaning cars at a parking lot near the San Diego International Airport and planned to graduate from O’Farrell Charter next year before joining the Marines, Garcia said. “We’re all in disbelief that this happened,” Garcia said through tears in a phone interview. According to his family, Gomez had headed out Friday with Suarez-Ojeda, who was dating Gomez’s sister. Joined by Said, the trio apparently made it to the barbecue in Ensenada, according to those who were there. But the San Diego teens did not make it back to the U.S. that night as expected. Said reportedly called his mother early Saturday and told her they were safe but had lost their cellphones, Garcia said. That was the last anyone heard from the group. Family members contacted Baja California authorities and frantically searched for the missing teenagers over the weekend, Garcia said. On Sunday night, Tijuana police told them about the three bodies found outside the Lomas Verdes apartment. “We were still holding out hope it wasn’t them,” Garcia said. But Gomez’s uncle went to a Tijuana morgue Monday and confirmed his nephew’s identity. Later that afternoon, news reached O’Farrell Charter about the gruesome slayings. “It’s been pretty devastating for students and staff,” Superintendent Jonathan Dean said Wednesday night. “It’s a tough situation.” Suarez-Ojeda and Gomez both began attending O’Farrell in the same year, when Suarez-Ojeda was in seventh grade and Gomez was in sixth grade. Four of Suarez-Ojeda’s siblings and Gomez’s younger brother still attend the schools. Dean and his staff made social workers and counselors available Wednesday. The high school’s senior class of about 135 students met after school Thursday to discuss plans for a memorial. The superintendent described Gomez as a “really nice, good kid” who was a leader this year during the football team’s inaugural season. John Van Houten, a former teacher and coach at O’Farrell who taught basketball, said Gomez was a fan of the sport, with many friends on the school’s basketball team. “He’d come to all our games,” Van Houten said. Gomez also played with them during lunchtime. “He was so hard working, always helping his parents out,” Garcia said of her cousin, who she described as being more of a little brother. “Nobody had a bad thing to say about him, he was just so goofy, happy and always smiling. “I know how this story sounds, like something that people hear on the news: You go to Tijuana and this happens,” Garcia said. “But he was the most selfless, kindest boy ever … This shouldn’t have happened to him.” Suarez-Ojeda had a more difficult road through school, dropping out at one point, Dean said. The teen’s mother was instrumental in convincing him to enroll at Ingenuity, an independent study program that shares a campus with O’Farrell, to get his high school diploma. He graduated last year. Suarez-Ojeda also played on the school soccer team, Van Houten said. Said attended Jose Vasconcelos high school in Tijuana and worked part-time as a barista at a coffee shop, his uncle said. Said was earning money for his own car and hoped to go to college, possibly to be an industrial engineer. “His name Angel was what he was to all his family,” Contreras said. “A great kid with good grades in school, kind, lovable. He will be missed.” He said the family believes the murders were a case of mistaken identity. “His whole family is broken,” Contreras said. Mexican officials said they are collaborating with U.S. authorities on the investigation. “When we have this type of incident, we work together with U.S. authorities,” Alvarez said. “They help us and we help them.” A spokeswoman for the FBI in San Diego said the agency was monitoring the situation. Faculty at O’Farrell set up a GoFundMe site for both San Diego families, and Garcia also set up a GoFundMe to help her cousin’s family, who she said was struggling financially even just to bring his body back to San Diego. GoFundMe has verified that the funds raised will go directly to the intended recipient. What does verified mean? (KGTV) - Three San Diego residents were shot to death during a weekend of violence in Tijuana. Multiple Tijuana-based news services said a triple homicide was reported on the morning of Nov. 25 in front of a building in the Lomas Verdes housing complex. When the bodies were found, they each had gunshot wounds to the head. According to news outlet Punto Norte, investigators believe the three people killed were taken from an apartment at gunpoint, forced to kneel and then shot. 10News spoke to 23-year-old Katheryn Garcia. She told us her cousin, 17-year-old Christopher Alexis Gomez was among the three people shot to death. Garcia said her cousin is a senior at O'Farrell Charter School (OCS) in Encanto. She said he was a proud starting Lineman for the school's new football team. Last week, Gomez told Garcia that he was going to a barbecue party in Ensenada on Friday with his friend, 18-year-old Juan Suarez Ojeda. Dr. Jonathan Dean, the Superintendent at OCS, told 10News Ojeda wasa recent O'Farrell Charter graduate, class of 2018. Garcia said she has never met the third victim, but knew the teen was Ojeda's friend who lived in Tijuana. Garcia said the three were tortured, likely in a different location, before being shot execution style. Garcia said her family are now left with more questions than answers. Why would Christopher, a boy who had never been in trouble, be killed? Was this a case of mistaken identity? Was he at the wrong place at the wrong time? Who would do this? “They’re monsters. Who does that to three young boys who had their whole life ahead of them," Garcia said in tears. The school set up a GoFundMe page to help Gomez’s and Suarez-Ojeda’s families with funeral costs and other expenses. Further details on the deaths on the San Diegans’ deaths were not released by authorities or news outlets. Four other deadly shootings occurred in the city during the weekend, according to Tijuana news reports.

Summary: Christopher Alexis Gomez and Juan Suarez-Ojeda crossed the US-Mexico border the day after Thanksgiving to attend a barbecue in Ensenada. Their plan was to head home to San Diego later that night. When they didn't return, family members contacted authorities in Baja California and searched for the pair over the weekend. On Sunday, Tijuana police gave them devastating news: The bodies of 17-year-old Gomez, 18-year-old Suarez-Ojeda, and an 18-year-old friend from Tijuana were found at an apartment complex in that city, the San Diego Union-Tribune reports. Local authorities say the three teens had been tortured, stripped down to their underwear, forced to kneel, and then shot in the head execution-style. Their bodies were discovered before the front door of an apartment on Lomas Verdes Avenue, local news outlet Punto Norte reports, adding that no shell casings were left behind and no one has been able to describe the killers. "Who does this to three young boys who had their whole life ahead of them?" Katheryn Garcia, Gomez's cousin, tells ABC10. Gomez was reportedly a football player and senior at O'Farrell Charter High School who had recently gotten his first job and was planning to join the Marines upon graduation. Suarez-Ojeda was a recent graduate of Ingenuity Charter School. Garcia tells the Union-Tribune that it was Gomez's first trip to Baja California, saying, "I know how this story sounds... you go to Tijuana and this happens. But he was the most selfless, kindest boy ever. This shouldn't have happened to him." In all, there were seven shooting homicides in Tijuana over that weekend, the Spanish-language Uniradio Informa reports. The O'Farrell Charter school has set up a GoFundMe page to help cover funeral expenses for the San Diego teens.

multi_news

en

Summarize: By. Daily Mail Reporter. and Ap. Vigilantes are searching an extensive cave system in the hunt for the last fugitive boss of the Knights Templar drug cartel, a "self-defense" group leader in western Mexico said Thursday. Vigilante leader Estanislao Beltran said there are signs the cartel had used the caves near the town of Arteaga in Michoacan state. 'I think they used it as a hideout,' said Beltran, who had descended about 100 meters (yards) into the caves. 'We found evidence.' Vigilantes: Groups of armed vigilantes search a cave for cartel leader Servando 'La Tuta' Gomez, a Mexican teacher-turned-drug kingpin. 'Self Defense': Groups of vigilantes calling themselves a'self defense' group stormed Gomez's hometown of Arteaga last week. Caves: The groups are searching an elaborate system of caves where Gomez is thought to be hiding out. Three days ago, vigilantes accompanied by federal forces took control of Arteaga, which is the hometown of Servando "La Tuta" Gomez, the only one of the cartel's top four leaders who has not been captured or killed. Gomez once served as a school teacher in the area and had some support among townspeople because his gang handed out money to residents. Beltran said some townspeople tried to prevent the vigilantes from entering Arteaga to hunt for cartel gunmen, as self-defense groups have down in more than a dozen townships in Michoacan over the last year. But he contended the demonstrators in Arteaga acted under threat from the cartel and said many people were becoming more trustful of the vigilantes. Residents want to get the local iron ore mines re-opened so the area's economy can start again. The government cracked down earlier this year on the Knights Templars' extensive involvement in iron ore exports to China. 'La Tuta': Servando 'La Tuta Gomez, a former teacher, is the alleged leader of the Knights Templar drug cartel. Signs: The vigilantes say that there are signs that the drug cartel has recently used some of the caves. Collaboration: The vigilantes team up with federal police officers in the search for 'La Tuta' The vigilantes sprang up in February 2013 to fight the cartel's extortion demands, kidnappings and killings. Armed with assault rifles, they travel from town to town in pickup trucks and set up highway checkpoints, seeking to expel cartel gunmen from the largely agricultural state. The government is trying to register and reign in the vigilantes, to avoid copy cats and infiltrators, like dozens of members of a rival cartel who were arrested earlier this week in Michoacan wearing vigilante-style T-shirts. The federal government has given the vigilante groups until May 10 to demobilize, which is usually understood as handing in their largest-caliber weapons and ending armed patrols. The government has offered them the option of signing up as members of an army-controlled Rural Defense corps. Robin Hood: Gomez has the support of many in his hometown of Arteaga because he often distributes money to the people living in the town. Last one: Gomez is the only remaining leader of the Knights Templar cartel, as the three others have either been captured or killed. Residents of Arteaga are accusing the man in the white shirt of being a supporter of the cartel, despite his claims siding with the vigilantes. Beltran said other self-defense fighters may become municipal policemen. Many of the state's local cops have been fired or re-assigned, in part because of suspected links to crime gangs. Interior Secretary Miguel Angel Osorio Chong appeared to suggest Thursday the demobilization would entail, as a minimum, that vigilantes leave their guns at home. 'Starting May 10, there cannot be armed people in the streets,' Osorio Chong told a news conference

Summary: Vigilantes last week stormed the Mexican town of Atreaga in search of Servando "La Tuta" Gomez. Gomez is the last remaining leader of the Knights Templar drug cartel who hasn't been either captured of killed. Prior to being a drug kingpin and international fugitive, Gomez was a school teacher. Gomez has the support of many of the residents of the town, as he often donates a lot of his money from the drug trade to the town and its people. The vigilante groups believe Gomez is hiding out in a complex system of caves on the outskirts of Arteaga.

cnn_dailymail

en

Summarize: Si è votato fino alle 15 di oggi lunedì 21 settembre per le elezioni regionali in Puglia, orario in cui sono state chiuse le urne per iniziare lo scrutinio delle schede. Sono già stati diffusi anche i primi exit poll per il voto pugliese mentre per ricevere i risultati di queste elezioni regionali 2020 in Italia, sarà necessario attendere la fine delle operazioni di scrutinio che si concentreranno dapprima sulle schede per il voto referendario sul taglio dei parlamentari per poi concentrarsi su quelle regionali. Testa a testa, dunque tra Emiliano e Fitto che restano i favoriti per la carica di Presidente della regione. L'affluenza registrata alla chiusura della giornata di ieri, domenica 20 settembre, ha fatto registrare una percentuale del 39,71% alle 23.00. La legge elettorale pugliese si basa su un turno unico con sistema proporzionale con premio di maggioranza. Gli elettori possono avvalersi del voto disgiunto ed esprimere due preferenze per candidati di genere diverso. Ecco tutte le informazioni per le regionali in Puglia, fino a che ora si vota oggi, i candidati, sono dicono i dati dei primi exit poll e quando arrivano i risultati definitivi. In Puglia si è votato ieri, domenica 20 settembre, dalle 7 alle 23, mentre oggi, lunedì 21 settembre, le urne sono rimaste aperte fino alle 15. Dopo la chiusura delle urne, sono iniziate le operazioni di scrutinio: l'ordine di spoglio darà precedenza alle schede referendaria, terminate le quali, si passerà allo scrutinio delle schede elettorali. Poco dopo le 15, però, sono arrivati i primi dati di exit poll e, circa un'ora più tardi, si avrà una prima proiezione. Per quanto riguarda i risultati definitivi su chi vincerà bisognerà aspettare la fine dello scrutinio. Gli ultimi sondaggi sulle intenzioni di voto in Puglia vedono il candidato del centrodestra Raffaele Fitto in leggero vantaggio sul governatore uscente Michele Emiliano; il primo totalizza tra il 39 e il 43% dei consensi, il secondo non va oltre il 40%. Molto più lontana la candidata del Movimento 5 Stelle Antonella Laricchia, che si attesta tra il 14 e il 18%. Alle 23 di domenica 20 settembre si è recato a votare per le regionali in Puglia il 39,71% degli aventi diritto, secondo i dati comunicati dal Ministero dell'Interno. Alle 19 di domenica 20 settembre il dato sull'affluenza era del 27,60%, mentre alle 12 si era fermato al 12,04%. L'affluenza definitiva alle elezioni di 5 anni fa fu del 51,1%. La legge elettorale che regola il voto alle regionali in Puglia è stata varata nel 2015 e prevede un sistema di voto proporzionale a turno unico; ciò significa che il vincitore è colui che riesce a ottenere anche un solo voto in più rispetto agli altri sfidanti. L'assegnazione dei seggi, che sono 50, viene portata avanti in parte su base circoscrizionale, in parte in base al listino unico regionale. Non c'è soglia di sbarramento ed è possibile effettuare il voto disgiunto, oltre che assegnare la doppia preferenza di genere. La scheda elettorale sarà di colore arancione, come si vede dal fac simile.

Summary: Urne chiuse anche per le regionali in Puglia dove alle 15 si sono chiusi i seggi per dare inizio alle operazioni di scrutinio. Si è conclusa dunque la tornata elettorale del 20 e 21 settembre e, appena dopo le 15, sono stati diffusi anche i primi exit poll. Particolarmente attesi i risultati della tornata elettorale pugliese dove la sfida principale resta quella tra Emiliano e Fitto. Ecco nel dettaglio tutte le informazioni sugli orari del voto, i primi exit poll e quando arriveranno i risultati per le regionali in Puglia.

fanpage

it

Summarize: Three men posed as police officers to force their way into a house and drag a father-to-be from his bed before they stabbed him to death in front of his pregnant fiancee, the Old Bailey heard. Danny Gough, 24, was dragged to the floor and stabbed so many times by Sam Monteith, Stephen Dougherty and Paul West, 30, it was impossible for him to survive. The court heard the three men had surveyed Mr Gough's fiancee's house for days and equipped themselves with an axe, machete and possibly a sword for the attack. Danny Gough, 24, was hacked to death in front of his pregnant fiancee after being dragged to the floor. They were the kind of weapons that could be used'silently but lethally', said Mark Heywood, prosecuting. The trio were caught after they left a Jiff lemon bottle, containing ammonia, behind which they used to spray in the eyes of Mr Gough so he could not protect himself. His pregnant fiancee Kareena Modashia was also sprayed so that she could not identify them. Mr Heywood said: ‘This case concerns a truly vicious killing, one motivated by vengeance and the need to remain in criminal control.' Mr Gough, 24, was stabbed with an axe, machete and possibly a sword until he was dead lying on the floor. The court heard Mr Gough had been involved in an ongoing feud with Stephen Dougherty, 36. Mr Gough's friend Leanne Meredith was involved in a fight with Dougherty and Monteith, 33, in a pub. Mr Gough took revenge by breaking into Monteith’s home in Wallington, Surrey, with two others. Mr Gough was hacked to death in front of his pregnant fiancee, Kareena Modashia, (left). A series of violent threats from the trio included having him raped in front of his mother Debbie Gough (right, arriving at the trial) Monteith was attacked with ammonia and knives, leaving a 10cm cut across his face - a scar clearly visible at court today during his trial. During the attack Mr Gough and the two others took Monteith’s phone, the court heard. In revenge Dougherty then called Mr Gough to make a series of threats in the weeks leading up to the killing. Paul West, 30, is accused of murdering Mr Gough. Dougherty told Gough he was going to have him raped ‘in front of your ugly mum,’ it is claimed. The men then carried out surveillance of Miss Modashia's house, watching neighbours and monitoring Mr Gough's condition so they could ensure the attack would occur when people were less alert. Mr Heywood said: ‘They had the information from someone close by, they dressed for the business and armed themselves with the kind of weapons which could be used silently but lethally, weapons with only one purpose - an axe, a machete and a long-bladed instrument, possibly a sword.’ The court heard the men then put their plan into action in the early hours of December 14 2010. Mr Heywood told the jury: ‘They went to his door, shouted police as a distraction, as a cover, they forced their way through the door and using information they went up the stairs and into the bedroom that he occupied with his girlfriend Kareena Modashia. ‘They dragged him from his bed, pulled him to the floor and hacked and stabbed at his body causing so many serious wounds that he had no hope of survival. ‘They left only when one of them said ‘he’s dead.' Mr Heywood said the trio then left quickly and quietly, disposing of anything that may link them to the killing. He added: ‘They left only one thing behind by mistake, it was dropped in the final moments when other members of the house heard and were awoken - it was a Jiff lemon bottle filled with ammonia. ‘It was used to spray into the eyes of the deceased so that, although he was sleeping and naked, he was unable to defend himself. ‘They also sprayed the others, his girlfriend and her mother so that it disabled them from identifying the attackers.' Dougherty, of no fixed address but formerly of Mijas, Spain; West, of Eaton Avenue, Slough, Berks, and Monteith, of Brighton Road, Sutton all deny murder. The trial continues. Sorry we are not currently accepting comments on this article

Summary: Danny Gough, 24, was dragged from bed and hacked with axe and machete. Sam Monteith, 33, Stephen Dougherty, 36 and Paul West, 30, deny murder. Old Bailey heard the trio surveyed Mr Gough's pregnant fiancee's house. They forced their way in, pretending to be police officers, and attacked him. Men sprayed ammonia into his fiancee's eyes so she could not identify them. Armed themselves with weapons that could be used'silently but lethally' Violent attack was revenge for ongoing feud and was a 'truly vicious killing'

cnn_dailymail

en

Summarize: AWAKENING When Siddhartha left the grove, where the Buddha, the perfected one, stayed behind, where Govinda stayed behind, then he felt that in this grove his past life also stayed behind and parted from him. He pondered about this sensation, which filled him completely, as he was slowly walking along. He pondered deeply, like diving into a deep water he let himself sink down to the ground of the sensation, down to the place where the causes lie, because to identify the causes, so it seemed to him, is the very essence of thinking, and by this alone sensations turn into realizations and are not lost, but become entities and start to emit like rays of light what is inside of them. Slowly walking along, Siddhartha pondered. He realized that he was no youth any more, but had turned into a man. He realized that one thing had left him, as a snake is left by its old skin, that one thing no longer existed in him, which had accompanied him throughout his youth and used to be a part of him: the wish to have teachers and to listen to teachings. He had also left the last teacher who had appeared on his path, even him, the highest and wisest teacher, the most holy one, Buddha, he had left him, had to part with him, was not able to accept his teachings. Slower, he walked along in his thoughts and asked himself: "But what is this, what you have sought to learn from teachings and from teachers, and what they, who have taught you much, were still unable to teach you?" And he found: "It was the self, the purpose and essence of which I sought to learn. It was the self, I wanted to free myself from, which I sought to overcome. But I was not able to overcome it, could only deceive it, could only flee from it, only hide from it. Truly, no thing in this world has kept my thoughts thus busy, as this my very own self, this mystery of me being alive, of me being one and being separated and isolated from all others, of me being Siddhartha! And there is no thing in this world I know less about than about me, about Siddhartha!" Having been pondering while slowly walking along, he now stopped as these thoughts caught hold of him, and right away another thought sprang forth from these, a new thought, which was: "That I know nothing about myself, that Siddhartha has remained thus alien and unknown to me, stems from one cause, a single cause: I was afraid of myself, I was fleeing from myself! I searched Atman, I searched Brahman, I was willing to dissect my self and peel off all of its layers, to find the core of all peels in its unknown interior, the Atman, life, the divine part, the ultimate part. But I have lost myself in the process." Siddhartha opened his eyes and looked around, a smile filled his face and a feeling of awakening from long dreams flowed through him from his head down to his toes. And it was not long before he walked again, walked quickly like a man who knows what he has got to do. "Oh," he thought, taking a deep breath, "now I would not let Siddhartha escape from me again! No longer, I want to begin my thoughts and my life with Atman and with the suffering of the world. I do not want to kill and dissect myself any longer, to find a secret behind the ruins. Neither Yoga-Veda shall teach me any more, nor Atharva-Veda, nor the ascetics, nor any kind of teachings. I want to learn from myself, want to be my student, want to get to know myself, the secret of Siddhartha." He looked around, as if he was seeing the world for the first time. Beautiful was the world, colourful was the world, strange and mysterious was the world! Here was blue, here was yellow, here was green, the sky and the river flowed, the forest and the mountains were rigid, all of it was beautiful, all of it was mysterious and magical, and in its midst was he, Siddhartha, the awakening one, on the path to himself. All of this, all this yellow and blue, river and forest, entered Siddhartha for the first time through the eyes, was no longer a spell of Mara, was no longer the veil of Maya, was no longer a pointless and coincidental diversity of mere appearances, despicable to the deeply thinking Brahman, who scorns diversity, who seeks unity. Blue was blue, river was river, and if also in the blue and the river, in Siddhartha, the singular and divine lived hidden, so it was still that very divinity's way and purpose, to be here yellow, here blue, there sky, there forest, and here Siddhartha. The purpose and the essential properties were not somewhere behind the things, they were in them, in everything. "How deaf and stupid have I been!" he thought, walking swiftly along. "When someone reads a text, wants to discover its meaning, he will not scorn the symbols and letters and call them deceptions, coincidence, and worthless hull, but he will read them, he will study and love them, letter by letter. But I, who wanted to read the book of the world and the book of my own being, I have, for the sake of a meaning I had anticipated before I read, scorned the symbols and letters, I called the visible world a deception, called my eyes and my tongue coincidental and worthless forms without substance. No, this is over, I have awakened, I have indeed awakened and have not been born before this very day." In thinking these thoughts, Siddhartha stopped once again, suddenly, as if there was a snake lying in front of him on the path. Because suddenly, he had also become aware of this: He, who was indeed like someone who had just woken up or like a new-born baby, he had to start his life anew and start again at the very beginning. When he had left in this very morning from the grove Jetavana, the grove of that exalted one, already awakening, already on the path towards himself, he had every intention, regarded as natural and took for granted, that he, after years as an ascetic, would return to his home and his father. But now, only in this moment, when he stopped as if a snake was lying on his path, he also awoke to this realization: "But I am no longer the one I was, I am no ascetic any more, I am not a priest any more, I am no Brahman any more. Whatever should I do at home and at my father's place? Study? Make offerings? Practise meditation? But all this is over, all of this is no longer alongside my path." Motionless, Siddhartha remained standing there, and for the time of one moment and breath, his heart felt cold, he felt a cold in his chest, as a small animal, a bird or a rabbit, would when seeing how alone he was. For many years, he had been without home and had felt nothing. Now, he felt it. Still, even in the deepest meditation, he had been his father's son, had been a Brahman, of a high caste, a cleric. Now, he was nothing but Siddhartha, the awoken one, nothing else was left. Deeply, he inhaled, and for a moment, he felt cold and shivered. Nobody was thus alone as he was. There was no nobleman who did not belong to the noblemen, no worker that did not belong to the workers, and found refuge with them, shared their life, spoke their language. No Brahman, who would not be regarded as Brahmans and lived with them, no ascetic who would not find his refuge in the caste of the Samanas, and even the most forlorn hermit in the forest was not just one and alone, he was also surrounded by a place he belonged to, he also belonged to a caste, in which he was at home. Govinda had become a monk, and a thousand monks were his brothers, wore the same robe as he, believed in his faith, spoke his language. But he, Siddhartha, where did he belong to? With whom would he share his life? Whose language would he speak? Out of this moment, when the world melted away all around him, when he stood alone like a star in the sky, out of this moment of a cold and despair, Siddhartha emerged, more a self than before, more firmly concentrated. He felt: This had been the last tremor of the awakening, the last struggle of this birth. And it was not long until he walked again in long strides, started to proceed swiftly and impatiently, heading no longer for home, no longer to his father, no longer back.

Summary: As he walks away from Govinda, Siddhartha realizes that he is embarking on a new stage of life. He has walked away from all his teachers, even Buddha, because they cannot teach the nature of the self. Siddhartha decides to learn from himself alone. As he walks, Siddhartha sees his surroundings as real and beautiful, rather than an illusion that causes suffering. For the first time, Siddhartha is experiencing the world on its own terms, rather than scorning what it has to teach him. This is his awakening. Siddhartha decides he has to start anew on his quest for enlightenment. Concurrent to this decision is the realization that he is completely alone. He has left his father, he has left the Samanas, and he has left Govinda with the Yellow-Robed Men. He can no longer define himself in relation to other men because he has no community.

booksum

en

Summarize: There are said to be around 6,500 spoken languages around the world. While 2,000 of these languages are spoken by fewer than 1,000 speakers, others are much more common with words and phrases being passed down through generations and shared across countries and continents. A map has been designed to highlight the spread of these common languages by plotting words, and their translations, from population to population. Type a word in the interactive map below. Called Word Map, the interactive guide (pictured) was created by Easyway Language Centre in Brazil. To use the map, type any word in any language into the search box and press Map It. As the words are mapped globally, the map reads each translation out loud. Coloured lines also connect countries with a common language. Called Word Map, the interactive guide was created by Easyway Language Centre in Brazil. If the tool above doesn't work in your browser, use the original version. To use the map, type any word in any language into the search box and press Map It. As the words are plotted globally, the map reads each translation out loud. Coloured lines also connect all the countries with a common language and where the word has the same translation. Clicking on a word reveals more about that particular country’s language, and how many countries share the translation, with details taken from Wikipedia. To use the map, type any word in any language into the search box and press Map It. As the words are mapped globally, the map reads each translation out loud. Coloured lines also connect all the countries with a common language and where the word has the same translation. Clicking on a word reveals more about that particular country’s language, and how many countries share the translation, with details taken from Wikipedia. In many instances, it shows how Latin formed the basis for modern-day languages, particularly in Europe. Typing the English word ‘cat’ reveals how many French speakers there are across the central band of countries in Africa, including Mali and Chad. In both Spanish and Portuguese, the translation is ‘gato’, and the coloured lines highlight how Spanish dominates South America, with the exception of Brazil. And how Portuguese is isolated in southern Africa in Mozambique. In terms of the number of speakers and dominance, Spanish is the most prominent language in Spain. Portuguese is universal in Portugal, but there are dialects in the south and central regions, including the dialect of Lisbon. The etymology of the word cat is said to come from a Late Egyptian word čaute, and was introduced in around the first century BC. Across Europe, 'cat', or 'catt', comes from the Latin 'cattus' and Byzantine Greek 'κάττα'. From here, it was introduced to many European languages. There is a noticeable difference in how the majority of Nordic countries write their translated version of 'cat', where the ‘c’ is replaced by a ‘k’, and how the word varies more across this region. Typing the English word ‘cat’ reveals how many French speakers there are across the central band of countries in Africa, including Mali and Chad. In both Spanish and Portuguese, the translation is ‘gato’, and this highlights how Spanish dominates South America, with the exception of Brazil. It also shows how Portuguese is isolated in southern Africa in Mozambique. The translations of the word ‘love’ are more varied. In French, Spanish and Italian, for example, it is shown as ‘aimer’, ‘amar’ and ‘amare’. It differs in Catalan, however, where it becomes ‘estimar. ’In Dutch, love translates as ‘liefde’, and German’s say ‘lieben.’ But further north it becomes ‘elsker’ and ‘alskar’. Meanwhile, in Russia as well as Belarus, Turkmeninstan and Ukraine, the word translates as ‘любовь’ For example, from the UK to the east, into Belgium, Germany, Bulgaria and Turkey the word evolves from ‘cat’ to ‘kat’ to ‘katze’ to ‘Котка’ to ‘kedi’ respectively. The map also shows the range of scripts in the Middle East and into Asia from Urdu to Hindi in India, Nepal’s Nepali and Thai. ‘Taxi’ is ‘taxi’ in many languages (Spanish highlighted.) It is shortened from ‘taximeter’, a tool used to measure distances and fares, and 'cabriolet' that refers to a horse-drawn carriage. Taximeter comes from Latin for ‘taxa’, meaning ‘tax’ The Word Map plots the official languages in each country, and doesn’t necessarily reflect the dominance of other languages in that region. For instance, Mandarin Chinese has the most native speakers around the world, but is only the official language of China and Taiwan, as well as one of four official languages of Singapore. Australia is listed as having no official language, although English is considered the as the national language of the country. Translations of the word ‘love’ are more varied. From the Old English ‘lufu’, the word has a Germanic origin, and from an Indo-European root it is shared with Sanskrit ‘lubhyati’ for ‘desires’, Latin’s ‘libet’, which translates into ‘it is pleasing’, as well as libido. While the Latin verb for ‘to love’ is 'amare'. These differences are seen in the way the word changes as the map moves around the world. In French, Spanish and Italian, for example, it is shown as ‘aimer’, ‘amar’ and ‘amare’. It differs in Catalan, however, where it becomes ‘estimar.’ In Dutch, love translates as ‘liefde’, and German’s say ‘lieben.’ But further north it becomes ‘elsker’ and ‘alskar’. Meanwhile, in Russia as well as Belarus, Turkmeninstan and Ukraine, the word translates as ‘любовь’ Of all the languages of Russia, Russian is the only official language. But, there are 35 different languages which are considered official languages in various regions of Russia, along with Russian. According to the map, Arabic is the official language of 21 countries, including Syria, Yemen and Oman. It belongs to the Semitic branch of the Afro-Asiatic family, and in this language and script the word science is translated into علم. The English word'science' originated in Middle English and has links with Old French and Latin ‘scientia’, from scire which means ‘know’. The noun 'ciencia' in Spanish translates to science, as well as erudition and knowledge. The English word'science' originated in Middle English and has links with Old French and Latin ‘scientia’, from scire which means ‘know’. The noun cicencia in Spanish translates to science, as well as erudition and knowledge. In German, the word translates to ‘wissenschaft’, where ‘wissen’ means knowledge and ‘schaft’ is a suffix used to denote that the word is feminine. In Finnish, science is ‘tiede’ The map also reveals that despite its vast size, the official language of the US, as well as Hawaii, is English, or American English. It isn't until Mexico (shown) that the word for'science', used as an example, changes into Spanish and 'ciencia' In German, the word translates to ‘wissenschaft’, where ‘wissen’ means knowledge and ‘schaft’ is a suffix used to denote that the word is feminine. In Finnish, science is ‘tiede’. On the map, Australia is listed as having no official language (pictured), although English is considered the as the national language of the country. The two ‘national’ languages of Finland are Finnish and Swedish. The official minority languages are three Sami languages, Romani, Finnish Sign Language and the Karelian language. Elsewhere, the word ‘internet’ is a portmanteau of the English words ‘inter’, for reciprocal or mutual, and ‘net’ for network. The technology was described in a paper in 1974, but the term is widely accepted as being coined in 1986 by the US Defense Department. Considered a proper noun by many, the word is often capitalised, but the term has become so widespread - and is often used interchangeably with the word web - that many people use it as a noun instead. As a result, the word hasn’t been translated globally, and as the map shows, is written as ‘internet’ in the various scripts. There are differences between the internet, and the web, namely that the internet is the network that connects computers and devices, while the web is used to move content around this network. However, the two terms are widely used to mean the same thing nowadays. Similarly, the English word ‘taxi’ is ‘taxi’ in many countries. This is because it shortened from ‘taximeter’ and ‘cabriolet’. The taximeter was invented in 1891 to measure distances and fares, while cabriolet refers to a horse-drawn carriage. Taximeter itself comes from Latin for ‘taxa’, meaning ‘tax’. The word ‘internet’ is a portmanteau of English words ‘inter’, for reciprocal or mutual, and ‘net’ for network. Considered a proper noun by many, the word is often capitalised, but the term has become so widespread - and is often used interchangeably with the word web - that many people use it as a noun instead. As a result, the word hasn’t been translated globally, and as shown, is written as ‘internet’ in various scripts

Summary: The map was created by Brazil-based Easy Way Language Centre and shows the official languages of countries. Type a word in any language into Word Map to hear it being translated globally in these official languages. As the words are mapped globally, lines connect all the countries with a common language. These lines in turn reveal how language has spread with migration and colonisation. Clicking a word reveals more about that particular language, and how many countries share the translation.

cnn_dailymail

en

Summarize: He shot and killed Brandon Lee on the set of the 1993 movie directed by Alex Proyas. Michael Massee, the actor who unfortunately will forever be known as the man who accidentally shot and killed Brandon Lee on the set of the 1994 film The Crow, has died. He was 64. Massee's death was first announced by actor Anthony Delon and confirmed to The Hollywood Reporter by a representative of his agent at Greene & Associates in Los Angeles. He and Delon recently worked together on the French TV series Interventions. His wife, Ellen, told The New York Times that Massee died Oct. 20 of stomach cancer in Los Angeles. In just his second movie appearance, Massee was playing a character known as Funboy when he fired a revolver that had been improperly prepared by crewmembers at Brandon Lee, the son of famed martial arts star Bruce Lee. The round was live and struck Lee in the stomach, and he died after several hours of surgery on March 31, 1993. He was 28. Massee was just following the script during filming at a studio in Wilmington, N.C. Shaken by the incident, he took a sabbatical from acting that lasted more than a year. “I don’t think you ever get over something like that,” he said. Most of The Crow, directed by Alex Proyas, had been completed before the tragedy, and the film was released in May 1994. It grossed more than $50 million in the U.S. Massee later played Ira Gaines, an American mercenary and former Navy SEAL, on the first season of the Fox drama 24 and portrayed Charles Hoyt, the scalpel-wielding killer known as the Surgeon, on the TNT crime drama Rizzoli & Isles. He also appeared as Gustav Fiers, aka The Gentleman, in The Amazing Spider-Man (2012) and its 2014 sequel. A native of Kansas City, Mo., Massee worked opposite Bill Pullman in two films released in 1997, David Lynch's Lost Highway and Wim Wenders' The End of Violence, and in the 2005 NBC miniseries Revelations. His film résumé also included the David Fincher films Seven (1995) and The Game (1997) as well as One Fine Day (1996), Amistad (1997), The Florentine (1999), Corky Romano (2001), Catwoman (2004) and The Amazing Spider-Man 2. On television, Massee also could be spotted on shows including The X-Files, Murder One, Alias, Cold Case, Supernatural, Flashforward, House and The Blacklist. Michael Massee, a prolific actor with nearly 80 screen credits in film and television between 1991 and 2016, has died at age 64, a representative at his talent agency, Greene & Associates, confirmed to EW. Massee died of cancer on Oct. 20 at Cedars-Sinai Medical Center in Los Angeles, though news of his death did not break until Wednesday morning. Massee is perhaps best known for his guest-starring role across 12 episodes of 24‘s first season in 2001, during which he played Ira Gaines, a leader of a terrorist cell. He also appeared on several episodes of HBO’s Carnivàle from 2003-2005 before taking parts on ABC’s Alias and Flashforward, The CW’s Supernatural, the NBC miniseries Revelations, and TNT’s Rizzoli & Isles. Massee was the actor who pulled the trigger on a faulty prop gun that accidentally killed Brandon Lee during production of a scene on 1994’s The Crow. The Kansas City, Missouri, native also appeared in several major films, including 2004’s Catwoman, 2012’s The Amazing Spider-Man, and the latter’s 2014 sequel. As news of Massee’s death broke, his peers took to Twitter to mourn their former collaborator. My heart is heavy to hear of his passing. He was IMMENSELY talented & had the kindest soul. I am privileged to have known him. #RIPMichaelM💔 https://t.co/QxcaGHcQOf — Angie Harmon (@Angie_Harmon) October 26, 2016 “My heart is heavy to hear of his passing,” wrote Angie Harmon, who starred opposite Massee on Rizzoli & Isles. “He was IMMENSELY talented & had the kindest soul. I am privileged to have known him.” Jon Cassar, one of 24’s executive producers, echoed Harmon’s sentiment. “Sad to hear of the passing of Michael Massee who played Ira Gaines, 24‘s first villain in season #1 RIP,” he tweeted. Sad to hear of the passing of Michael Massee who played Ira Gaines, 24's first villain in season #1 RIP pic.twitter.com/MBT2uw3ZD3 — Jon Cassar (@joncassar) October 26, 2016 Massee is survived by his wife, Ellen, whom he married in 1997, and two children, Lily, 17, and Jack, 13. Uploaded by samiam78blm on From 2005 - Actor Michael Massee talks about the accident that occurred on the set of The Crow in 1993, that killed Brandon Lee. Once and ONLY time he has commented on accidentally killing Brandon Lee on the set of The Crow. I do not approve idiotic responses, these include; hateful comments about Lee or Massee, murder theories, or unfactual rants. If your grasp of English and grammar is poor, save yourself the time writing comments. If I cannot understand your comment, it is NOT getting approved. http://www.brandonleemovement.com Michael Massee, the character actor sadly remembered for his role in the accidental death of Bruce Lee’s son, has died at the age of 61. Massee’s death was reportedly first announced by actor Anthony Delon — who recently costarred with Massee on the French TV series Interventions — and later confirmed by The Hollywood Reporter. The details of his death were not disclosed. While acting in just his second feature film, the cult-classic The Crow, Massee fired a faulty prop revolver at the film’s star, Brandon Lee, the son world-famous actor and martial artist Bruce Lee. Unbeknownst to Massee, the gun fired a live round, striking Lee in the stomach. He died after several hours of surgery on March 31, 1993, at the age of 28. After years of silence, Massee finally opened up about the incident in a 2007 interview with Extra. “It absolutely wasn’t supposed to happen. I wasn’t even supposed to be handling the gun until we started shooting the scene and the director changed it.” After the incident, Massee said, “I just took a year off and I went back to New York and didn’t do anything. I didn’t work. What happened to Brandon was a tragic accident … I don’t think you ever get over something like that.” Fortunately, most of the film was completed before the accident, and it was released in May 1994, grossing more than $50 million domestically. Massee is also known for playing Ira Gaines, the mercenary villain on the first season of Fox’s 24. He also played a villain known as the Surgeon, or Charles Hoyt, on the TNT’s Rizzoli & Isles. He also appeared as Gustav Fiers, or The Gentleman, in 2012’s The Amazing Spider-Man and its 2014 sequel. A native of Kansas City, Missouri, Massee also appeared in films like Seven (1995), The Game (1997), One Fine Day (1996) and Amistad (1997). Massee TV credits included The X-Files, Murder One, Alias, Cold Case, Supernatural, Flashforward, House and The Blacklist.

Summary: Michael Massee, who accidentally shot and killed Bruce Lee's son, Brandon Lee, on the movie set of The Crow in 1993, has died at the age of 61, reports the Hollywood Reporter. The actor died of cancer in a Los Angeles hospital last Wednesday, his talent agency tells Entertainment Weekly. Massee, a native of Kansas City, Mo., was appearing in only his second movie when he fired a prop revolver at Brandon Lee while filming Alex Proyas' The Crow in Wilmington, NC. Unbeknownst to Massee, crew members had mistakenly loaded the gun with real bullets. Lee was hit in the stomach and died on March 31 after hours of surgery. "It absolutely wasn't supposed to happen. I wasn't even supposed to be handling the gun until we started shooting the scene and the director changed it," Massee said in a 2005 interview, per People. "I don't think you ever get over something like that." Massee took more than a year off from acting following the incident but went on to appear in movies Se7en, Lost Highway, The End of Violence, The Florentine, Corky Romano, Catwoman, The Amazing Spider-Man, and in TV shows 24, Rizzoli & Isles, The X-Files, Alias, Cold Case, House, and more. He leaves behind a wife and two children, aged 13 and 17.

multi_news

en

Summarize: By. Michael Zennie. PUBLISHED:. 21:01 EST, 25 October 2012. |. UPDATED:. 22:34 EST, 25 October 2012. Busted: Michael R Mastro had been hiding out in the French Alps with his wife Linda for 16 months. A bankrupt Seattle real estate magnate and his wife have been arrested in France more than a year after allegedly skipping out on $250million in debts. Michael and Linda Mastro, age 87 and 63, were arrested on Wednesday at the idyllic Lake Annecy in the French Alps - where they had been hiding out after their $2billion real estate empire fell apart. The couple fled after refusing to hand over a pair of massive diamond rings Mrs Mastro owned. The rings were 27.8 carats and 15.9 carats and worth more than $1.4million combined. They were found in the small village of Doussard in eastern France near the Swiss border, where they had been renting a house under their own name for 16 months, the Seattle Times reports. U.S. authorities are currently working to extradite them back to Seattle. They face six counts of federal bankruptcy fraud as part of the biggest bankruptcy case in Washington state history. In 2009, Mr Mastro claimed $587million in debts and $250million in assets. That debt is estimated at $250million today. In the heyday of their real estate business, the Mastros lived on top of the world. The couple owned a Bentley convertible, a Rolls Royce, a Land Rover and a Mercedes-Benz. They vacationed in Italy, Switzerland, France, New York, and Palm Springs, Florida. They bought expensive works of art, including dozens of sculptures by renowned artist Dale Chihuly. When their real estate empire collapsed, federal authorities say, the couple tired to hide the assets from creditors, according to the Times. Wealthy: Mr Mastro's Bently convertible was among the luxury items auctions off in a feeble attempt to clear his $250million debts. Hanging on: This is one of the massive diamond rings that the Mastros refused to give up. They allegedly set up shell companies in Belize and the state of Delaware and transferred ownership of their home in California, as well as their cars and Mrs Mastro's expensive jewelry. Even after they filed bankruptcy, they continued to drive luxury cars and take lavish vacations to resorts all over the world, the Times reports. In June 2011, after a judge ordered them to turn over the lavish rings, they moved out of their expansive home in Palm Desert California and fled. Federal authorities say they missed the couple in Toronto, Canada, by just days. And then, for 16 months, they disappeared. In their absence, many of their belongings were sold at auction, in an effort to cover their debts. Among the estate was a Chihuly chandelier, a Steinway baby grand piano, 56 designer handbags. 65 pairs of high heels and a Bentley convertible. The total from that sale was $341,000. In January, a judge ordered $2.8million from the Mastro's assets to be distributed to creditors -- just one penny on the millions owed by the couple. A world away: The Mastros spent 16 months on the run from federal authorities. They were found in the French Alps. Valuable: This is just one of the works of art by renowned glass artist Dale Chihuly that the Mastros owned

Summary: Michael and Linda Mastro declared bankruptcy in 2009, with $587million in debts and $250million in assets. They were found living in a house rented under their own name on Lake Annercy in the French Alps. Michael Mastro had a $2billion real estate empire in Seattle, Washington, before the collapse. Couple are accused of trying to use shell companies to Belize and Delaware to hide the assets from creditors. Analysts expect investors to get back just pennies for every dollar invested.

cnn_dailymail

en

Summarize: By. Michael Seamark. PUBLISHED:. 20:51 EST, 6 November 2012. |. UPDATED:. 09:01 EST, 7 November 2012. An MP last night accused Channel 4 of commissioning a ‘loaded’ documentary on the Leveson Inquiry into Press standards that will be presented by Hugh Grant. Alun Cairns has written to its chief executive, David Abraham, asking how a programme fronted by the actor ‘will ever comply with Ofcom’s rules about impartiality’. Mr Grant, who has become the public face of the Hacked Off pressure group demanding Press reform, is making the documentary. Scroll down for video. MP Alun Cairns questioned whether a documentary on the Leveson Inquiry presented by Hugh Grant, who fronts the Hacked Off campaign demanding Press reform, can comply with Ofcom's impartiality rules. At Leveson: Celebrities Steve Coogan, Hugh Grant and Max Mosley pictured at the Inquiry into Press Standards. The production company, Films of Record, was founded by Roger Graef, who is its chief executive. He was, until yesterday, also chairman of the Media Standards Trust, which founded Hacked Off. In his letter, which has been copied to Ofcom chief Ed Richards, Tory MP Mr Cairns says he is ‘extremely concerned’ about the programme. He details the links between Mr Grant, the production company and the MST which, the politician points out, has ‘produced a series of recommendations for a system of statutory regulation of the Press which Hacked Off and Hugh Grant support’. Mr Cairns continues: ‘The celebrity spokesman of Hacked Off making the case for the implementation of the MST’s proposals is none other than Hugh Grant – the chosen celebrity presenter of the programme. It is little wonder why he accepted the very kind invitation from Films of Record to front the programme.’ On the campaign trail: Actor Hugh Grant, pictured with Lib Dem Dr Evan Harris, right, and journalist John Kampher, left, at a Hacked Off meeting. Campaign: Hugh Grant said that Government intervention is necessary on the Andrew Marr show. The MP for the Vale of Glamorgan says he has seen an email from the producer of the programme which states: ‘The film follows Hugh Grant as he investigates what forces may be ranged against his campaign to make the Press more accountable…’ Mr Cairns says he suspects Mr Graef’s resignation as head of the Trust is ‘an attempt to hide conflicts of interest which brought this programme about’. Mr Cairns quotes Ofcom’s Broadcasting Code which states broadcasters ‘should not give undue prominence to the views of particular persons or bodies on matters of political or industrial controversy and matters relating to current public policy’. The MP wants Mr Abraham to answer:. Celebrity status: Mr Cairns said the matter is too important to depend on celebrity support. Last night Mr Cairns said: ‘It’s important that we consider the Leveson Inquiry objectively. It’s too important to depend on celebrity support. The issue goes to the heart of journalism and uncovers some of the most difficult issues.’ A Channel 4 spokesman said: ‘The programme will be impartial in line with Ofcom’s Broadcasting Code.’ The MST website yesterday announced: ‘Helena Kennedy QC is temporarily taking over the Chair of the Media Standards Trust, as Roger Graef, the founder of Films of Record, has stepped aside due to broadcasting commitments. ‘Ten Alps/Films of Record has been commissioned to make a programme about the Leveson Inquiry, and although Roger Graef is not involved, he wants no suggestion that there may be any conflict of interest with the Media Standards Trust.’ VIDEO: Hugh Grant talks about what he thinks is of public interest at the Leveson inquiry

Summary: MP for the Vale of Glamorgan Alun Cairns asks how a show fronted by the actor can 'ever comply with Ofcom's rules about impartiality' Mr Cairns voiced concerns in letter to C4 chief executive David Abraham. How the owner of a production company lobbying for a particular outcome can be allowed to air a programme supportive of his own agenda. What investigations C4 has made into the funding of the MST, Films of Record, Hacked Off and Mr Grant. How a celebrity spokesman for a lobbying campaign can present a programme in an impartial way. How the owner of a production company can lobby on the one hand for one organisation's success, then present an impartial programme about that.

cnn_dailymail

en

Summarize: Background CID carries out IRS’ criminal law enforcement responsibilities under three principal statutes. Under title 26 U.S.C., IRS has authority to investigate alleged criminal tax violations, such as tax evasion and filing a false tax return. Under title 18 U.S.C., IRS has authority to investigate a broad range of fraudulent activities, such as false claims against the government and money laundering. Under title 31 U.S.C., IRS is responsible for enforcing certain recordkeeping and reporting requirements of large currency transactions, such as cash bank deposits of more than $10,000. In carrying out its responsibilities, CID coordinates as necessary with IRS’ District Counsel, the Tax Division within the Department of Justice (Justice), and local U.S. Attorneys to prosecute violators of these statutes. Combating money laundering and other financial crimes is considered a high priority by both Justice and the Department of the Treasury (Treasury). According to Justice officials, because of CID’s expertise in conducting detailed financial investigations, U. S. Attorneys and law enforcement agencies routinely rely on CID’s assistance in investigating financial crimes, particularly those involving money laundering related to narcotics trafficking. In addition, CID agents have access to tax information, which they can use to develop financial investigations more fully. CID is also involved in ongoing efforts to identify and investigate emerging financial crimes, such as health care and bankruptcy fraud. According to CID officials, although such assistance places competing demands on CID’s time, it also aids in establishing the cooperative environment conducive to getting CID’s tax cases prosecuted and in obtaining information that can lead to criminal tax investigations. Historically, CID’s total staffing and budget have represented about 5 percent of IRS’ overall resources. As of the end of fiscal year 1996, CID had the full-time equivalent of 4,504 staff, including 3,065 special agents, and a budget of $366 million. CID is also reimbursed for some of the assistance it provides to other law enforcement agencies, particularly Justice’s Organized Crime Drug Enforcement Task Force (OCDETF) program. Scope and Methodology To determine the actions CID has taken since the early 1990s to increase the time spent on tax investigations relative to nontax investigations, we interviewed senior CID officials in IRS’ National Office, officials from both the Tax and Criminal Divisions in Justice, and officials from the Office of the Under Secretary for Enforcement in Treasury. We interviewed the CID officials because of their responsibilities in managing CID operations and setting division policies. Justice and Treasury officials were interviewed to obtain their opinions regarding CID’s (1) assistance in narcotics and money laundering investigations and (2) increased emphasis on tax investigations. We also reviewed CID’s annual goals and objectives and annual performance reports for fiscal years 1990 through 1996, as well as relevant documentation on the reorganization of its administrative functions and operations. To determine the types of investigations initiated and the results of referrals to U.S. Attorneys for prosecution and sentences resulting from these prosecutions, we analyzed IRS’ Criminal Investigation Management Information System (CIMIS) data. CID uses CIMIS data to track the status and overall results of its criminal investigations, including the direct investigative time (DIT) expended on investigations. DIT is the amount of time that CID agents spend directly working on investigations. We selected fiscal years 1990 through 1996 to identify CID’s investigative trends in order to capture data from the time of the IRS studies that raised concerns about CID’s investigative priorities through fiscal year 1996, the most recent period for which data were available. We obtained and analyzed CIMIS data to identify nationwide by fiscal year (1) the number and results of various types of CID investigations, and (2) the principal sources of information that led to CID investigations. CID staff, at our request, reconfigured CIMIS data for fiscal years 1990 through 1996 to reflect the current IRS field alignment of 4 regions and 33 districts, as well as CID’s current program areas—fraud and narcotics—with fraud further broken out between tax gap fraud and other fraud. Other than reconciling the totals from CIMIS data extracts to CID annual performance reports, we did not verify the accuracy of the CIMIS data. We did our work from October 1996 to August 1997 in accordance with generally accepted government auditing standards. The work was done at IRS’ National Office and Southeast Regional Office; IRS’ Georgia, South Florida, and Delaware-Maryland District Offices; and at U. S. Attorney’s offices in the Northern District of Georgia, the Southern District of Florida, and the Maryland District. We selected the offices we visited because of the proximity of our staff working on this assignment. We requested comments on a draft of this report from the Acting Commissioner of Internal Revenue and the Attorney General. Their comments are discussed at the end of this letter. Actions to Increase Direct Investigative Time Spent on Tax Investigations In the early 1990s, concerns raised in IRS studies regarding CID’s investigative priorities spurred CID to take actions to increase the amount of time its agents spent on tax investigations. Between fiscal years 1990 and 1992, the percent of DIT spent on tax gap investigations decreased from 56 percent to 46 percent; since then, the percent of DIT spent on tax gap investigations increased to 59 percent as of fiscal year 1996. CID has established a range of 57 to 61 percent of DIT to be spent on tax gap investigations as its goal for fiscal year 1997 and beyond. Subsequent to hearings on IRS employee misconduct in 1989 before the Subcommittee on Commerce, Consumer and Monetary Affairs, House Committee on Government Operations, the Commissioner of Internal Revenue appointed an independent panel to review various concerns raised during the hearings, including issues relating to criminal investigations. In its October 1990 report, the panel stated that there had been a significant decrease in CID resources applied to tax investigations and a corresponding increase in resources applied to nontax investigations. The panel believed that CID’s work priorities were not properly aligned with IRS’ strategic goal of increasing taxpayers’ compliance with the tax laws. The panel recommended that CID (1) establish a criminal enforcement policy in line with IRS’ overall efforts to increase compliance with the tax laws, (2) ensure that its allocation of resources and mix of cases are consistent with such a policy, and (3) closely monitor and control implementation of this policy through the National Office. Also, to address concerns about whether CID’s workload was properly balanced between tax and nontax investigative efforts, IRS convened a study group that included representatives from Justice and Treasury. The study group’s August 1991 report found that CID resources used for tax investigations had declined about 18 percentage points between fiscal years 1980 and 1990; on that basis, the study group recommended that resources devoted to tax investigations be increased and that future resources devoted to narcotics investigations be limited to the amount expended in fiscal year 1991. The IRS Executive Committee agreed with these recommendations, and in June 1992 CID initiated an action plan to implement them. The actions led to a reorganization of CID, which began in October 1993 and was fully implemented in October 1994. The reorganization was done in part with the intent of giving CID’s national office a better means to control and monitor field activities to keep them aligned with national policies and objectives as recommended by the review panel and the study group. In terms of its organizational structure, CID was reduced from 7 regions and 63 districts to 4 regions and 34 districts. In addition, the position of Director of Investigation (DI), reporting directly to the IRS National Office Assistant Commissioner for Criminal Investigation, was established in each region to oversee and coordinate investigative activities. The DIs replaced seven former Assistant Regional Commissioners for Criminal Investigation, who reported directly to the Regional Commissioners. The DIs are responsible for ensuring that CID field offices adhere to national office program objectives and policies. Another action CID took to better track the allocation of its resources to tax versus nontax investigations was to consolidate its major program areas and to establish a specific category for tax gap investigations. In fiscal year 1995, CID consolidated the five program areas under which investigations had been categorized and tracked—narcotics, organized crime, public corruption, financial compliance, and other illegal crime—into two principal program areas—fraud and narcotics. The fraud program was subdivided into tax gap fraud and other fraud. Tax gap fraud pertains to investigations of legal industries with alleged criminal tax violations. The other fraud category involves investigations of illegal industries or money laundering investigations with no tax-related charges.The narcotics program primarily relates to investigations of money laundering activity by individuals and organizations involved in narcotics trafficking. Beginning in fiscal year 1996, CID set specific national goals for the percent of DIT to be used on tax gap and narcotics investigations to help ensure that additional resources would be allocated to tax gap investigations. The fiscal year 1996 DIT goals were 58 percent for tax gap investigations—1 percent higher than the actual DIT for fiscal year 1995—and 24 percent for narcotics investigations—the same as the actual DIT for fiscal year 1995. Since CID began taking these actions, DIT applied to tax gap investigations has increased. (See fig. 1.) According to data provided by IRS, the percent of time applied to tax gap investigations for fiscal years 1993 through 1996 increased 13 percentage points. As of fiscal year 1996, CID applied 59 percent of DIT to tax gap investigations, exceeding its goal by 1 percentage point, while applying 22 percent of DIT to narcotics investigations, 2 percentage points short of its goal. According to CID national office officials, the goal for the amount of DIT to be allocated to tax gap investigations in fiscal year 1997 is a range of 57 to 61 percent, and the goal for narcotics investigations is a range of 23 to 25 percent. They stated that these goals, which are expected to be the goals for the next few years, were developed with input from the DIs. It is CID officials’ judgment that these goals will enable CID to (1) conduct investigations in support of IRS’ strategic goal of increasing compliance with the tax laws; (2) contribute to the government’s efforts in combating narcotics and money laundering; and (3) continue allocating some of its investigative time to cases involving emerging financial crimes, such as health care fraud. Trends in the Number and Results of CID Investigations for Fiscal Years 1990 Through 1996 CID considers completed investigations that merit referral to the U.S. Attorneys for prosecution as an important step toward the eventual prosecution, conviction, and sentencing for criminal tax violations and related financial crimes. By publicizing convictions, CID hopes to deter others from engaging in such criminal activity and to promote voluntary compliance with the tax laws. Consequently, CID uses statistical data from CIMIS to track the number and percent of investigations initiated, as well as the number and percent of referrals made to U.S. Attorneys for prosecution and sentences handed down by the U.S. courts based on CID cases. CIMIS data show that the percent of tax gap investigations initiated, the percent of tax gap cases referred to U.S. Attorneys for prosecution, and the percent of court sentences based on tax gap cases have all begun to increase since CID increased the time spent on tax gap investigations. However, as of fiscal year 1996, the increases have not been enough to match fiscal year 1990 levels for these indicators. Trends in the Types of Investigations Initiated As shown in figure 2, tax gap investigations represented 54 percent of all CID investigations initiated in fiscal year 1996. This is an increase over the fiscal year 1992 level of 47 percent and just under the fiscal year 1990 level of 55 percent. The figure also shows that between fiscal years 1990 and 1996, narcotics investigations decreased from 30 percent to 25 percent of all CID investigations initiated. Other fraud investigations were 6 percentage points higher in fiscal 1996 than in fiscal year 1990. Trends in Investigations Referred to the U.S. Attorneys for Prosecution Figure 3 shows that, in general, the percent of CID cases being referred to the U.S. Attorneys for prosecution for tax gap fraud since fiscal year 1992 has increased, while the percent of other types of referrals—narcotics and other fraud cases—either declined or remained somewhat stable. Specifically, tax gap referrals represented 47 percent of all CID referrals in fiscal year 1996 compared to 39 percent in fiscal year 1992 and 49 percent in fiscal year 1990. Trends in Sentences From Cases Referred to the U.S. Attorneys for Prosecution Court sentences—including incarceration, probation, and fines—based on tax gap investigations decreased from 54 percent of all court sentences based on CID investigations in fiscal year 1990 to a low of 37 percent in fiscal year 1994. Since that time tax gap sentences have increased to 44 percent of all court sentences for CID cases as of fiscal year 1996. Overall, sentences based on narcotics investigations increased from 32 percent to 39 percent of all court sentences based on CID investigations between fiscal years 1990 and 1993, then decreased to 31 percent as of fiscal year 1996. (See fig. 4.) Additional information relating to CID’s investigations between fiscal years 1990 and 1996 is shown in appendixes I, II, III, and IV. Appendix I shows the number of staff days applied nationwide by type of criminal investigation. Appendix II contains information on the percent of DIT applied to each type of criminal investigation by IRS location. Appendix III shows the numbers of investigations, referrals to the U.S. Attorneys for prosecution, and court sentences by type of criminal investigation. Appendix IV discusses the principal sources of information on which CID’s investigations were based. Agency Comments and Our Evaluation IRS and the Department of Justice each provided comments on a draft of this report. Each agency generally agreed with the information presented in the report and offered technical comments, which we have incorporated where appropriate. Copies of this report are being sent to the Chairmen and Ranking Minority Members of the Senate Committee on Finance, the Senate Committee on Governmental Affairs, the House Committee on Ways and Means, and the House Committee on Government Reform and Oversight; the Chairman and Ranking Minority Member of the Subcommittee on Treasury, General Government, and Civil Service, Senate Committee on Appropriations; and the Chairman and Ranking Minority Member of the Subcommittee on Treasury, Postal Service, and General Government, House Committee on Appropriations; various other congressional committees; the Secretary of the Treasury; the Attorney General; and other interested parties. We will also make copies available to others upon request. Major contributors to this report are listed in appendix VI. Please contact me on (202) 512-9110 if you or your staff have any questions about this report. Investigative Time Applied Nationwide by Type of Criminal Investigation in Fiscal Years 1990 Through 1996 This appendix presents the nationwide number of staff days applied directly to the major types of Criminal Investigation Division (CID) investigations. This does not include staff days applied to noninvestigative activities, such as training. Table I.1: Number of Direct Staff Days Applied Nationwide by Type of Criminal Investigation in Fiscal Years 1990 Through 1996 Number of staff days by fiscal year Note 1: Investigative time spent following up on information provided to CID that indicates potential criminal violations prior to initiation of an investigation is categorized as information items. Note 2: Totals do not add due to rounding. Percent of Direct Investigative Time Applied to CID Investigations in Fiscal Years 1990 Through 1996 This appendix shows the percent of direct investigative time (DIT) applied to the major types of CID investigations nationwide, by regions, and by district offices from fiscal years 1990 through 1996. For fiscal year 1996, CID set national DIT goals of 58 percent for tax gap investigations and 24 percent for narcotics investigations. To achieve these goals, CID requested that the Directors of Investigation for each region help to ensure that the regional DIT was within a range of 58 to 60 percent for tax gap investigations and 24 to 26 percent for narcotics investigations. Table II.1: Percent of Total DIT Applied to Tax Gap Investigations by Location in Fiscal Years 1990 Through 1996 Percent of DIT by fiscal years (continued) Percent of DIT by fiscal years Table II.2: Percent of Total DIT Applied to Narcotics Investigations by Location in Fiscal Years 1990 Through 1996 Percent of DIT by fiscal years (continued) Table II.3: Percent of Total DIT Applied to Other Fraud Investigations by Location in Fiscal Years 1990 Through 1996 Percent of DIT by fiscal years (continued) The Number and Results of CID Investigations in Fiscal Years 1990 Through 1996 This appendix presents information on the number of CID investigations that were initiated, the number of investigations in which CID recommended prosecution, and the number of sentences resulting from prosecutions from fiscal year 1990 through fiscal year 1996. According to CID officials, completing an investigation and subsequently prosecuting and sentencing the subject of the investigation may take a year or more. As a result, the number of prosecution recommendations and sentences shown in a particular fiscal year in tables III.2 and III.3 may not have resulted from the investigations initiated in the corresponding fiscal year in table III.1. Sources of Information That Resulted in CID Investigations in Fiscal Years 1990 Through 1996 CID relies on various sources of information for initiating its investigations, including information from (1) within IRS, such as from the Examination Division; (2) other government sources, such as U.S. Attorneys; (3) currency transaction reports; and (4) the public. Although information from other government sources may result in various types of CID investigations, including tax fraud, information from within IRS predominantly results in tax fraud investigations. Information provided from within IRS and by other government sources accounted for about 75 percent of the total CID investigations initiated each year for fiscal years 1990 through 1996. As shown in figure IV.1, investigations based on information provided by other government sources increased from about 45 percent in fiscal year 1990 to about 51 percent in fiscal year 1996. Investigations based on information provided from within IRS fluctuated from about 32 percent in fiscal year 1990 to about 29 percent in fiscal year 1996. In 1991, a study group convened to examine CID’s workload and to recommend changes to better balance, direct, and strengthen its future investigative activities recommended that IRS reemphasize its internal fraud referral program. In an effort to increase the quality of internal fraud referrals from other IRS groups to CID that may lead to tax fraud investigations, IRS established formal fraud referral procedures effective for fiscal year 1996. This included establishing the position of fraud coordinator in each district office to act as a focal point for fraud referrals. According to CID officials, the objective of these procedures is to increase coordination between CID and other IRS functions, particularly the Examination Division, in an effort to ensure that only cases involving potential criminal fraud, rather than civil fraud, are referred to CID. CID officials further stated that it is too soon to determine the overall success of the new procedures. Comments From the Internal Revenue Service Major Contributors to This Report General Government Division, Washington, D.C. Atlanta Field Office A. Carl Harris, Assistant Director Clarence Tull, Senior Evaluator Sally P. Gilley, Evaluator The first copy of each GAO report and testimony is free. Additional copies are $2 each. Orders should be sent to the following address, accompanied by a check or money order made out to the Superintendent of Documents, when necessary. VISA and MasterCard credit cards are accepted, also. Orders for 100 or more copies to be mailed to a single address are discounted 25 percent. U.S. General Accounting Office P.O. Box 37050 Washington, DC 20013 Room 1100 700 4th St. NW (corner of 4th and G Sts. NW) U.S. General Accounting Office Washington, DC Orders may also be placed by calling (202) 512-6000 or by using fax number (202) 512-6061, or TDD (202) 512-2537. Each day, GAO issues a list of newly available reports and testimony. To receive facsimile copies of the daily list or any list from the past 30 days, please call (202) 512-6000 using a touchtone phone. A recorded menu will provide information on how to obtain these lists.

Summary: GAO reported on the actions the Internal Revenue Service's (IRS) Criminal Investigation Division (CID) has taken since the early 1990s to increase the time spent on tax investigations versus nontax investigations, focusing on: (1) investigations initiated by CID; and (2) referrals to U.S. Attorneys for prosecution and court sentences based on these investigations, for fiscal year (FY) 1990 through FY 1996. GAO noted that: (1) between FY 1990 and FY 1992, IRS data show that the percent of time spent on tax gap investigations decreased by 10 percentage points, continuing a downward trend since the early 1980s; (2) on the basis of the recommendations of two IRS studies done in the early 1990s, CID began in October 1993 taking actions designed to increase the amount of time its agents spent conducting tax investigations: (3) specifically, CID reorganized its administrative functions and operations with the intent of better targeting resource allocations; (4) it also consolidated and recategorized its program areas with an objective of better targeting its investigations; (5) in addition, as of FY 1996, CID established goals for the percent of time to be spent on its investigations, particularly for tax gap investigations; (6) since these actions were initiated, the percent of time spent on tax gap investigations has increased by 13 percentage points from a low of 46 percent in 1992 to 59 percent in 1996; (7) overall, the 59 percent in FY 1996 represented a net increase of 3 percentage points over the FY 1990 level; (8) between FY 1992 and FY 1996, there was an increase in the percent of tax gap investigations that CID initiated and in the percent of referrals to U.S. Attorneys for prosecution based on tax gap cases; (9) since FY 1994, the percent of court sentences based on tax gap cases has also increased; (10) however, as of FY 1996 the increases in these indicators have not been enough to match FY 1990 levels.

gov_report

en

Summarize: Allred to David Boreanaz: Pay Up or Else! Gloria Allred to David Boreanaz: Pay Up or Else! went public today with his extra-marital affair aftercontacted Boreanaz' lawyer and demanded six figures... sources tell TMZ.David admitted to PEOPLE mag he had a relationship with a woman ---- but "She asked for money. I felt as though I was being blackmailed...."TMZ knows what went down. We're told Boreanaz had a short-term relationship with the woman -- one source says they hooked up "2 or 3 times." Sources say he started paying her money -- several thousand dollars here and there -- but she began demanding more, threatening to go public with the affair. Boreanaz then confessed the affair to Jaime Bergman, his wife of 9 years.Enter Gloria Allred, who reps the mistress. Allred contacted Boreanaz' lawyer -- legal pit bull-- and demanded 6 figures.Boreanaz then blunted the attack by going public with his affair. He has refused to pay the mistress another cent.Gloria Allred just told TMZ what we told you yesterday... Rachel Uchitel was not the woman who tried getting $$$ out of Boreanaz. Bones star David Boreanaz admitted to an extra-marital affair on Monday and claimed his former mistress contacted an attorney and threatened to go to various media outlets. RadarOnline.com has confirmed that the woman is represented by attorney Gloria Allred. She strenuously denies any implication of impropriety on her part and puts the blame for his actions squarely on Boreanaz. Boreanaz did not name the woman he slept with but RadarOnline.com has learned exclusively that it was Tiger Woods’ mistress Rachel Uchitel! EXCLUSIVE REPORT: Rachel Uchitel Had An Affair With David Boreanaz And yes, Uchitel is represented by Allred. More bad news for Boreanaz: Another woman he slept with has also hired Allred, RadarOnline.com has confirmed. When Boreanaz’ slept with Uchitel his wife Jaime Bergman was pregnant at the time of the affair! PHOTOS: Tiger Mistress Rachel Uchitel In Bikini Yes, we’d love to take credit for this scoop but the affair was first reported by Star magazine way back when Tiger Woods still had a wholesome image! In a cover story, Star magazine blew the lid of Boreanaz’s affair with Uchitel. Shortly after that Star and the National Enquirer broke the story of Uchitel’s affair with Tiger. And shortly after that Uchitel hired Allred and scored a mega-million dollar payout from the golfer. PHOTOS: Celebrity Cheaters Now the Bones star says he’s concentrating on putting back together his nine year marriage. RadarOnline.com has learned that Boreanaz had ANOTHER affair and Allred represents the new woman too! PHOTOS: Rachel Uchitel at The Hamptons in 2007 So will he now confess to TWO affairs? “I was associated with a woman who I was involved with and had a relationship with,” Boreanaz says. “She asked for money. I felt as though I was being blackmailed or there was some sort of extortion.” PHOTOS: Rachel Uchitel On St.Barth With Ryan Seacrest So we now know definitively that two women who had extra marital affairs with Boreanaz are represented by Allred. So which client is she actively representing now in regard to Boreanz? The new, mystery woman, according to Allred, which makes us wonder if Boreanaz will now publicly admit to his affair with Uchitel now that he’s in a truth-telling mood. PHOTOS: Sexy Pics of All of Tiger’s Women And for the record, Allred told RadarOnline.com: ”It is a typical line of many male celebrities to make that claim (about blackmail). There is no merit to it in this case. I do represent a woman who had a relationship with him. Now that he is attacking her, I do expect that she will want to tell the story of their relationship so that the truth will come out.”

Summary: David Boreanaz admits he cheated on his wife, but what he didn't confess was that he cheated with none other than the original Tiger Woods mistress, Rachel Uchitel. If you were paying very close attention when the Tiger scandal broke, you might have already guessed-reports noted that Uchitel had been linked to Boreanaz as far back as October. So is she the alleged blackmailer? Boreanaz has another former mistress, Radar reports, and she's represented by the favored attorney of wronged women everywhere, Gloria Allred. Although Allred is representing Boreanaz's mystery woman-and reportedly asked Boreanaz's lawyer for a six-figure payout-TMZ reports Uchitel is not the woman doing the blackmailing.

multi_news

en

Summarize: Morning commuters were stunned to see a hot air balloon, which had been enjoying a scenic flight over Melbourne, descend in the middle of a car park on Monday. 'Light and variable winds' forced the pilot and his nine passengers to land outside Footscray Town Hall on Monday at around 7am, not far from a busy railway line. According to Mike, a caller on 3AW Radio who witnessed the landing, there was 'a mild scene of panic as crew and people who were on the balloon madly tried to pack up the balloon'. Onlookers were stunned when a red hot-air balloon landed in the empty car park at Footscray Town Hall on Hyde St in Melbourne's west on Monday morning at around 7am. According to Mike, a caller on 3AW Radio who witnessed the landing, 'a mild scene of panic as crew and people who were on the balloon madly tried to pack up the balloon' But Damian Crock, director of Picture This Ballooning, told Daily Mail Australia that the landing was 'routine' and safety was the pilot's priority. 'The winds became light and there was not much reliable steerage. Rather than using up lots of fuel, the pilot found a safe and available landing spot', he said. Mr Crock said the hour-long flight was close to its end when it landed in the car park. The balloon displaying the Fujitsu logo allegedly landed 25-metres away from a railway line connecting Footscray and Seddon train stations. A driver snapped this image moments before the Fujitsu balloon landed in the Footscray car park. Thankfully, no one was injured during the impromptu landing. 'Everyone had a ball and they all went back to breakfast at level 35 at the Sofitel Hotel. They all had a marvellous time', Mr Crock said. Action shots taken from onlookers show the red balloon making its way down from the sky to the empty car park, as the balloon slowly deflated to the ground. Action shots taken by onlookers show the red balloon making its way down to the empty car park. Picture This Ballooning has made around 15,000 landings in Melbourne since it was established 18 years ago. 'Annually we might get a couple of routine landings - they are quite uncommon'. Mr Crock said Melbourne was the first city in the world to begin flying commercial hot air balloons and now balloons fly nearly 150 days a year. Earlier this year, another Fujitsu balloon was forced to make an emergency landing in a home's front yard in Hawthorn, Melbourne. The pilot was forced to land after winds took control of the balloon, 3AW reported. The Fujitsu balloon was flying for nearly an hour before it made a 'routine landing'. Onlookers watched as the balloon slowly deflated to the ground. Thankfully, the pilot and his nine passengers were not injured during the impromptu landing

Summary: Picture This Ballooning's red hot air balloon made a 'precautionary landing' in the car park at Footscray Town Hall on Hyde St in Melbourne's west. The balloon displaying the Fujitsu logo was carrying 10 people, including the pilot, on Monday morning. After wind conditions became 'light and variable', the pilot found a safe spot to land. A witness told 3AW that there was 'a mild scene of panic' Damien Crock, director of Picture This Ballooning, said it was a 'routine landing' and 'everyone had a ball'

cnn_dailymail

en

Summarize: Introduction In 1956, the year Congress authorized the Interstate Highway System, there were 37,965 fatalities on U.S. roads—6.05 fatalities for every 100 million vehicle miles traveled (VMT). The construction of limited-access highways spurred travel by automobile, leading to an increase in the number of fatal accidents. Congress responded with a series of laws that have helped reduce the fatality rate by 80% over the past six decades. By 2014, the United States recorded only 1.08 fatalities for every 100 million VMT, although the rate ticked up to 1.13 per 100 million VMT in 2015 ( Table 1 ), and again in the first 9 months of 2016, when fatalities rose to 1.15 per 100 million VMT. Development of new motor vehicle technologies, investments in building safer highways and educating motorists, and improving emergency medical services all have contributed to reduced fatality rates. Congress has played a significant role in improving highway safety by directing the federal government to impose and enforce safety standards for motor vehicles. This effort has been at times controversial, and several large recalls have raised questions about the effectiveness of federal motor vehicle regulation. Federal Motor Vehicle Safety Standards In the early decades of the automobile, U.S. vehicles were lightly regulated by a combination of state and private-sector standards. National regulation was generally not seen as appropriate; in the early 1900s, according to two historians of auto safety, it was widely believed that "the only useful and politically acceptable action Congress might take was to help the states and localities construct more and better roads." The Society of Automotive Engineers (SAE), a professional association founded in 1905, became the primary source of vehicle safety rules for many decades. State governments often used SAE recommendations to set their own standards for vehicle brakes, headlamps, and windshield wipers. At the same time, the rising number of highway deaths prompted a new interest in vehicle safety: between 1962 and 1964, Congress passed three safety bills into law, including a seat belt regulation. The new laws were only a precursor to broader federal regulation. Two publications also spurred interest in a greater federal role. Ralph Nader's 1965 book, Unsafe a t Any Speed: The Designed-in Dangers of the American Automobile, argued that cars were unnecessarily unsafe and that the auto industry should be regulated by a federal agency. Also influential was Accidental Death and Disability: The Neglected Disease of Modern Society, a National Academy of Sciences report that documented the impact of accidental injuries, including those by motor vehicles. Comprehensive vehicle safety legislation was passed in the form of the National Traffic and Motor Vehicle Safety Act of 1966. As approved unanimously by both houses of Congress and signed by President Lyndon B. Johnson, the legislation had two parts: 1. The Highway Safety Act of 1966 mandated that each state put in place a highway safety program in accordance with federal standards to improve driver performance, accident records systems, and traffic control. 2. The National Traffic and Motor Vehicle Safety Act of 1966 directed the Secretary of Commerce (later changed to the Secretary of Transportation when that agency was established in 1967) to issue safety standards for all motor vehicles beginning in January 1967. A National Traffic Safety Agency was established to carry out the provisions of the new law; it was renamed the National Highway Traffic Safety Administration (NHTSA) in 1970. Since its establishment, NHTSA has issued dozens of safety standards, including regulations affecting windshield wipers, hood latches, tires, brakes, seat belts, and airbags. Proposing and finalizing a NHTSA safety regulation can take many years: all NHTSA regulations follow the Administrative Procedure Act of 1946 (APA), which ensures that proposed rulemaking is publicized in the Federal Register, comments are taken and considered, and agency decisions are clearly explained. Court review of standards is allowed, and revisions to federal regulations must also follow the APA. NHTSA does not verify in advance that motor vehicles and parts comply with its standards. Instead, the law provides that "[a] manufacturer or distributor of a motor vehicle or motor vehicle equipment shall certify to the distributor or dealer at delivery that the vehicle or equipment complies with applicable motor vehicle safety standards prescribed under this chapter.... Certification of a vehicle must be shown by a label or tag permanently fixed to the vehicle...." Manufacturers are responsible for testing their vehicles and are liable for recalls and penalties if they are later found not to meet NHTSA's Federal Motor Vehicle Safety Standards (FMVSS). After a new model goes on sale, NHTSA buys a sampling from dealers and tests the vehicles at its own facilities to determine whether they comply. If NHTSA determines there is noncompliance, it can encourage the manufacturer to recall the model to correct the problem, or it can order a recall. In addition to promulgating motor vehicle standards and addressing vehicle defects, NHTSA's mission also includes providing assistance to states on traffic safety issues, such as drunk driving and distracted driving, and maintaining a comprehensive database about motor vehicle crashes. Estimates of Effects of Federal Safety Standards A recent NHTSA study estimated that passenger vehicle safety technologies associated with Federal Motor Vehicle Safety Standards (FMVSS) have saved 613,501 lives between 1960 and 2012. The study evaluated the effects of 31 motor vehicle technologies mandated by NHTSA, including dual master cylinders and front disc brakes, electronic stability control, energy-absorbing steering assemblies, seat belts, door locks, airbags, and side door beams. It estimated that the risk of a fatality in 2012 was 56% lower than in 1960, based on evaluation of the effectiveness of specific technologies in reducing occupant fatalities. The NHTSA report found seat belts, introduced in the late 1960s, to have been responsible for more than half of all the lives saved, 329,715, and that their effectiveness rose sharply after NHTSA required installation of combined lap and shoulder belts in place of simple lap belts in 1974. However, the study also highlighted the importance of other measures in addition to federal vehicle safety regulation: it estimated that the number of lives saved annually by seat belts rose from 800 to 6,000 after many states allowed police to issue tickets if a driver or passengers were not wearing seat belts. Every state but New Hampshire has enacted laws requiring seat belt use. The study notes that the full benefits of new federal safety standards may take many years to be felt. The passenger vehicle fleet turns over slowly; nearly half the cars and light trucks on the road are more than 12 years old. And standards can take many years to develop and issue. Although electronic stability control was introduced as standard equipment on one make of vehicle in 1998 and was subsequently adopted on some other makes, only 22% of light vehicles on the road were equipped with the technology in calendar year 2012. FMVSS required electronic stability control to be included in all new vehicles starting in model year 2012. The study estimates that more than 1,362 lives may be saved annually when all vehicles on the road utilize the technology, but this will not occur for a couple of decades. In a separate study in 2012, NHTSA evaluated the crashworthiness and crash avoidance performance of passenger cars and light vehicles, isolating the vehicle element in traffic safety improvements from human and environmental effects. The study did not focus solely on FMVSS-regulated technologies, but also included overall vehicle design and improvements initiated by manufacturers. Unlike NHTSA's Lives Saved by Vehicle Safety Technologies and Associated Federal Motor Vehicle Safety Standards, 1960 to 2012, this study did not address specific technology and product sources of the improvements. The NHTSA report found that the likelihood of crashing in 100,000 miles of driving had decreased from 30% in a new model year 2000 vehicle to 25% in a new model year 2008 vehicle. The likelihood of escaping a crash uninjured improved from 79% to 82% in the same time period. The report contended that "the nationwide impact of these advancements is substantial" and that vehicle improvements between 2000 and 2008 prevented 700,000 vehicle crashes, prevented (or mitigated) injuries of 1 million occupants, and saved 2,000 lives in calendar year 2008 alone. Trends in Vehicle Recalls In addition to promulgating and enforcing vehicle safety standards, NHTSA investigates vehicle defects that affect safety. NHTSA's Office of Defects Investigation (ODI) r eviews and investigates comp laints of alleged defects from vehicle owners, a utomakers, and other sources. There are several routes a potential recall complaint can take: Denial. When NHTSA's analysis of petitions calling for defect investigations leads the agency to decide not to proceed, it publicizes the reasons for the denial in the Federal Register. Further R eview. If NHTSA determines there is reason to open an investigation of alleged safety -related defects, it looks further into the facts and ends with either a recommendation that the manufacturer recall the vehicle or a determination that there is no safety-related defect. If a safety defect is confirmed by NHTSA, most manufacturer s will initiate a recall ; if they fail to do so, NHTSA can initiate a recall itself. In addition to the NHTSA investigative process, manufacturers also conduct their own in ternal in vestigation s; if a manufacturer find s that a vehicle or component does not comply with a federal safety standard, it may issue its own recall to c orrect a safety defect before accidents are reported. The law establishing the motor vehicle safety program requires that a manufacturer of a defective vehicle or component notify the vehicle owner and fix the defect without charge. In practice, most recalls are issued by manufacturers, sometimes influenced by a NHTSA defect finding and sometimes solely by a manufacturer upon its own finding of a defect. Of the 1,039 recalls issued in 2016, 92 were issued by manufacturers influenced by a NHTSA finding, and 947 were issued based on a manufacturer's finding alone. The annual number of recall action s has generally risen in the past decade (except for the recession year of 2009), and the number of vehicles and items of equipment recalled has risen steeply since 2013 ( Figure 1 ). There are several reasons for the rising number of recalls, including stricter laws, larger fines, delayed detection by NHTSA of vehicle problems, and several recent high-visibility cases affecting millions of vehicles. In 2014 and 2015, two large recalls were issued: General Motors (GM) recalled 2.2 million vehicles because of faulty ignition switches, which could slip out of the "run" position and prevent air bags from deploying in crashes. GM acknowledged that the defective switches caused 15 deaths and a number of injuries. NHTSA assessed a maximum $35 million civil penalty against GM. I n a separate settlement, the Department of Justice fined GM $900 million in criminal penalties.Nineteen manufacturers recalled a total of about 42 million vehicles due to a defect in airbags provided by Takata, a parts supplier. The defect may cause the airbags' inflators to explode. The faulty airbags are linked to 16 deaths globally. In the United States, there have been 220 cases of Takata-supplied airbag inflators exploding, with 11 deaths and 184 injuries. NHTSA fined Takata $200 million, with $70 million due in cash; an additional $130 million payment would be demanded if Takata fails to meet its commitments or if additional violations of the law are determined. Separately, the Department of Justice fined Takata $1 billion, including a $25 million criminal fine, $125 million for victim compensation, and $850 million for compensating automakers for a portion of the cost of recalling the vehicles. In response to the GM ignition switch recall, NHTSA evaluated its procedures, interactions, and communications with General Motors. Its publication NHTSA's Path Forward in 2015 outlined how the lessons learned from that recall could improve its defect investigation system. The then-NHTSA Administrator, Mark Rosekind, wrote in that publication that it is no overstatement to say [the ignition switch recall] was one of the most significant cases in NHTSA's history, not only because of the tragic toll of deaths and injuries, or the technical challenges it presented, but because of the unprecedented steps the manufacturer took to conceal a deadly defect. Path Forward identified five shortcomings in the recall and NHTSA procedures that affected its handling of the GM ignition switch problem: 1. GM withheld critical information about engineering changes that would have allowed NHTSA to more quickly identify the defect. 2. NHTSA did not hold GM accountable for providing inadequate information. 3. Neither GM nor NHTSA completely understood the application of advanced airbag technology in GM vehicles. 4. NHTSA did not consider alternate theories proposed by internal and external sources. 5. NHTSA did not identify and follow up on trends in its own data sources and investigations. The report proposed various process improvements, including increasing auto industry accountability, increasing NHTSA's knowledge of emerging technologies, and improving defects investigations. Why Have Recalls Increased? In addition to high-profile cases involving millions of vehicles, four other factors may have changed the magnitude of motor vehicle recalls. These are described below. Trends in M anufacturing E fficiency. Motor vehicle manufacturers are attempting to reduce the number of separate parts they use by installing a single part on multiple vehicle models, instead of designing unique parts for each model. For example, Ford is cutting its global platforms from 15 to nine. Much of the auto industry's sourcing is global, and one effect of having fewer vehicle platforms may be that a defective part is installed in a very large number of vehicles sold under several brands. The defective Takata airbags, for example, were used by nearly every automaker, leading to recalls in other countries as well as the largest recall on record in the United States. Stricter F ederal R eporting R equirements and S tiffer P enalties. More thorough and earlier reporting requirements and steeper penalties are thought to have increased the number of defects reported and hence the number of recalls. In 2000, after a highly publicized recall of Ford Explorer sport utility vehicles and the Firestone tires used on those vehicles, Congress passed the Transportation Recall Enhancement, Accountability, and Documentation Act (TREAD Act). The law established an Early Warning Reporting System (EWRS) that requires vehicle manufacturers to report a wide range of information, including data on defects, injuries, and deaths related to use of their products, enabling NHTSA to investigate defects without waiting for complaints from vehicle owners. In addition, the law increased civil penalties for violations of safety standards from a maximum of $925,000 to $15 million and provided criminal penalties for misleading NHTSA about safety defects that cause death or injury. NHTSA issued final TREAD Act regulations in 2003. EWRS regulations were not followed by manufactures in some recent recalls, however, leading NHTSA to impose additional penalties. I nadequate D ata, A nalysis, and T raining at NHTSA. Some recalls might be smaller if they were identified earlier. NHTSA's Office of Defects Investigation (ODI) is responsible for identifying and investigating potential vehicle safety issues and requiring recalls when warranted. In June 2015, the Department of Transportation Inspector General (DOT OIG) made 17 recommendations to improve NHTSA's procedures for collecting and analyzing vehicle safety data and deciding when to investigate. That report states that weakness in ODI's training and supervision of pre-investigation staff and its processes for identifying potential safety concerns and initiating investigations, as evidenced by NHTSA's handling of the GM ignition switch defect, deter NHTSA from successfully meeting its mandate to help prevent crashes and their attendant costs, both human and financial. DOT OIG notes that "without detailed guidance, decisions regarding key aspects of early warning reporting data are left to manufacturers' discretion—resulting in inconsistent reporting and data that ODI investigative chiefs and vehicle safety advocates consider to be of little use." While the DOT OIG has found that NHTSA has made "considerable progress" in addressing these recommendations, it told the Senate Committee on Commerce, Science, and Transportation in November 2016 that five recommendations remain open: four that will improve early warning reporting data and an improvement in the consumer complaint quality control process. The 2015 surface transportation bill, the Fixing America's Surface Transportation (FAST) Act, tied an increase in NHTSA's funding authorization to DOT certification that the inspector general's 17 recommendations had been implemented. Agency Funding. The Obama Administration requested additional funding for NHTSA. In June 2015, the NHTSA Administrator testified on behalf of the agency's budget request: Fixing problems such as the Takata recalls and Fiat Chrysler's recall performance is a monumental task. Yet the agency must manage this enormous and necessary task with its existing people, technology, and authorities. NHTSA must accomplish this task with a defects investigation budget of $10.6 million, a figure that, when adjusted for inflation, is actually 23 percent lower than its budget 10 years ago. The President has submitted a budget request that would fund significant improvements in NHTSA's defect investigation efforts.... In light of the DOT OIG report, however, the Commerce Committee opted to tie additional funding to the resolution of those issues. Chairman John Thune spoke about the committee's perspective when the surface transportation bill was discussed on the Senate floor: [T]he Obama administration claimed NHTSA's problems could be solved by simply throwing more money at the agency, but based on the expert testimony from the inspector general, it is clear money alone is not going to solve the problem. We need to ensure that the agency fixes what is broken before we provide a significant increase in funding authorization with taxpayer dollars. Recall Completion Rates Remain an Issue It is rare that all owners of a recalled vehicle bring their vehicles to a dealer for repairs. As a result, many defective vehicles are still on the road long after a recall is initiated. A recent review of NHTSA data by J.D. Power and Associates, a market research company, found that of the more than 120 million vehicles recalled from 2013 through 2015, 45 million had not been repaired as of mid-2016. Big recalls have the lowest completion rates: recalls affecting fewer than 10,000 vehicles have a 67% completion rate, while recalls affecting more than a million vehicles have a completion rate of 49%. The J.D. Power report suggests that bigger recalls are more complicated: manufacturers have more difficulty locating all owners. Obtaining an adequate supply of replacement parts can delay repairs. In addition, owners of vehicles involved in smaller recalls are easier to contact through personalized communication methods, such as a phone call. The J.D. Power report also found that vehicle age, vehicle type, and the nature of the safety issue affected recall completion rates. Newer vehicles (model years 2013-2017) were completed at a 73% rate; older vehicles (model years 2003-2007) had a 44% completion rate. This may have reflected the difficulty of identifying the owners of vehicles that were more than six years old at the start of the period J.D. Power studied. The highest completion rates were for recalls involving powertrain, hydraulic brakes, and electrical issues: 71%, 66%, and 62%, respectively. By comparison, 47% of airbag issues and 48% of suspension problems were fixed. In a separate, earlier study, the U.S. Government Accountability Office (GAO) reviewed vehicle recalls for the period 2000 through 2008 and found that the average completion rate in those years was 65%. GAO's analysis found a wide differential among automakers: some had completion rates as low as 23%, while others had rates as high as 96%. Some manufacturers had consistently higher or lower rates. GAO called for NHTSA to implement changes that could improve the defect recall process, including adopting additional defect notification methods; modifying defect notification letters; better publicizing existing resources, such as the NHTSA website, and including a Vehicle Identification Number (VIN) search engine on the NHTSA website; and developing national standards that would categorize the severity of a recall and whether a vehicle should be operated. As discussed later in this report, the FAST Act mandated that NHTSA and manufacturers develop new approaches to reach out to owners of recalled vehicles. New Technology and Vehicle Safety Many new technologies, whether mandated by Congress or NHTSA or developed by automakers, have translated incrementally into safer motor vehicles. As the introduction of new vehicle technologies has accelerated in the past decade, moving toward much more vehicle automation and a long-term goal of a fully autonomous vehicle, Congress and federal regulators are grappling with how to encourage such advancements, while recognizing that the traditional regulatory process is long and could "stymie innovation and stall the introduction of these technologies." A range of new technologies are being introduced to motor vehicles, many of them bringing automation to vehicular functions once performed only by the driver. Mary Barra, chairman and CEO of General Motors, has observed that "the auto industry will change more in the next five to 10 years than it has in the last 50." There are three forces driving motor vehicle innovation: technological advances enabled by new materials and electronics; consumer demand for telecommunications connectivity and new types of vehicle ownership and ridesharing; and regulatory mandates pertaining to emissions, fuel efficiency, and safety. Technological Advances Most technological advances evolve from earlier technologies. For example, cruise control, a mechanism that takes over the throttle of the car to maintain a steady speed set by the driver, was invented in 1948 and first used on vehicles 10 years later. It has developed into a more automated function called adaptive cruise control, which automatically adjusts vehicle speed to maintain a safe distance from vehicles ahead. Several such innovations are expected to improve driver and passenger safety in the coming years. These include the technologies described below. Antilock Brake Systems (ABS) ABS were originally invented for use on aircraft, but by the 1990s had been modified for use on automobiles. Today they are a standard feature being used as a base for further technological advances, as described below. ABS prevent the wheels from locking up during hard braking or on slippery surfaces (such as an icy road). Sensors at each wheel and a computer interact to maximize braking and prevent lock-up. Traction Control and Electronic Stability Control (ESC) Traction control is an electronically controlled system that limits wheel spinning during acceleration. Using the antilock braking system, traction control brakes a spinning wheel and automatically shifts power to the opposite drive wheel. ESC is an advanced form of this system that brakes the wheels and keeps the vehicle on the driver's intended path. Automatic Emergency Braking (AEB) or Brake Assist The AEB system detects a sudden effort to stop the car and, working with ABS, applies the brakes to reach the shortest stopping distance. By 2020, some vehicles may have driver override systems with sensor technology that will apply the brakes if a crash is imminent, even if the driver is pressing the accelerator. Forward-Collision Warning (FCW) FCW uses cameras, radars, and lasers to search for cars ahead of a vehicle and alerts drivers if they are heading for an imminent crash with another vehicle, using visual signals and sounds to alert the driver. A similar system—pedestrian detection—is available to detect a pedestrian in the vehicle's path. Blind-Spot Warning (BSW) Radar or cameras prompt a device on an outside mirror to light up if another vehicle is in the driver's blind spot, preventing an accident. Advanced BSW may also include devices that steer a vehicle back to the center of a lane if another vehicle is detected in a blind spot. Lane Departure Warning (LDW) The system works by using cameras or lasers to monitor lane markings and sending visual or audible signals to a driver or vibrating the steering wheel or seat if the vehicle leaves its lane, unless a turn signal is activated. Lane-keeping assist takes LDW one step further and activates a sensor that will correct the steering direction. The Insurance Institute for Highway Safety (IIHS) has found that drivers who fall asleep, suffer a medical emergency, or black out from drug or alcohol use are most likely to veer out of their intended lane. Lane departure is one of the major reasons for highway fatalities. Single-vehicle crashes where vehicles leave the road accounted for 40% of fatal crashes in 2014; head-on collisions and sideswipes (which also can be caused at times by lane departures) account for another 12% of the fatal crash total. Active Head Restraints In a crash, the force of a driver or passenger in a front seat activates sensors that automatically move the head restraint forward to firmly cushion the occupant's head and reduce whiplash, which is a major consequence of such crashes. Automatic High Beams This technology automatically switches headlights from low to high beam and back, depending on road visibility. Biometric Vehicle Access Most automakers are moving away from key-based vehicle access, replacing it with electronic keyless entry systems. This links vehicle access to electromagnetic frequency and communication wavelengths that may leave the vehicle subject to hacking. In the future, biometric technology may eliminate this risk by unlocking a vehicle only with biometric identification, such as a fingerprint. Telematics Drivers or passengers can use telematics—a combination of telecommunications with information and communications technology—to communicate with a central dispatch center or 911 emergency call center using cellular telephone and Global Positioning Satellite (GPS) technologies. The vehicle location is transmitted and, if airbags deploy, emergency service can be notified. These telematics can also be used as Remote Vehicle Shutdown to immobilize stolen cars. Automated Vehicles Increasingly, such innovations are being combined as manufacturers produce vehicles with higher levels of automation. Some envision a day when vehicles will be fully automated, with little or no involvement of the human passengers. With each level of automation, it is forecast that crashes may be dramatically reduced. Vehicles do not fall neatly into two categories of automated and nonautomated, because all of today's motor vehicles have some element of automation. The Society of Automotive Engineers International (SAE), a 100-year-old international standards-setting organization, has developed six categories of vehicle automation, a classification that has also been adopted by NHTSA to foster standardization and clarity in discussions about growing vehicle automation and safety ( Table 2 ). Consumer Demand Motor vehicles and consumer electronics are increasingly connected. A sign of this transformation is seen in the annual Consumer Electronics Show (CES), which now serves as a showcase for automakers' near-term and future vehicle models. Vehicles began using more electronics in the 1990s with telematics and infotainment. As more sensors, cameras, and telecommunications features, including Internet, are added to vehicles, consumer digital technology is becoming one of the driving forces of motor vehicle innovation. These new systems provide consumers with vehicles with capabilities for entertainment and navigation assistance ; convenience through easier entry, ignition, and phone mobility; greater comfort through suspension adjustment, brake assist, and cabin temperature control; as well as more security through ABS, blind-spot detection, and 911 crash notification. A survey by the Boston Consulting Group (BCG) shows that consumers seek digital innovations in vehicles: when considering purchase of a new car, U.S. consumers said that connectivity and safety are ranked in the top five of new features. The same survey showed that consumers under 30 years of age value digital-device integration in vehicles. A McKinsey & Company report, which forecasts motor vehicle revenues between 2015 and 2030, shows growth of vehicle and aftermarket sales in those years. However, the largest sales increases are forecast in on-demand, shared mobility services, such as car sharing, and in vehicle-related data-connectivity, including remote services and software upgrades. Regulatory Mandates Emission, fuel economy, and vehicle safety regulations are a third factor increasing the demand for more technologically advanced vehicles. In the past decade, hybrid and electric vehicles have established a beachhead, while internal combustion engines—which are forecast to remain dominant in passenger motor vehicles for many decades—have been retooled so that their fuel economy has increased and emissions have dropped. Plug-in electric new vehicle sales have grown from just over 17,000 units in 2011 to nearly 160,000 units in 2016 (out of total U.S. passenger and light-truck sales in 2016 of 17.6 million vehicles). Sales grew by 37% in 2016 when compared to 2015. While many electric vehicles are purchased by "early adopters" who want to experience this type of relatively new technology, state and federal emissions and fuel economy rules also play a part. More than half of the new plug-ins sold in 2016 were sold in California, influenced by the state's zero-emission vehicle (ZEV) mandate, which requires that a certain percentage of an automaker's sales must be ZEVs (electric and fuel cell vehicles). California has established a goal of placing 1.5 million ZEVs on its highways by 2025. The Obama Administration's greenhouse gas (GHG) emissions program—a joint regulatory initiative of NHTSA and the Environmental Protection Agency (EPA)—seeks reductions in GHG emissions and an increase of vehicle fuel economy (to 54.5 miles per gallon by model year 2025). In announcing the program in 2012, the Obama White House noted the expected technology-enhancing effects of the program: [A]chieving the new fuel efficiency standards will encourage innovation and investment in advanced technologies that increase our economic competitiveness and support high-quality domestic jobs in the auto industry. [M]ajor auto manufacturers are already developing advanced technologies that can significantly reduce fuel use and greenhouse gas emissions beyond the existing model year 2012-2016 standards. In addition, a wide range of technologies are currently available for automakers to meet the new standards, including advanced gasoline engines and transmissions, vehicle weight reduction, lower tire rolling resistance, improvements in aerodynamics, diesel engines, more efficient accessories, and improvements in air conditioning systems. President Trump announced in Detroit in March 2017 that his Administration will review the GHG emissions program, which may lead to a change in the emissions and fuel economy standards. The convergence of a high level of motor vehicle industry innovation, consumer choice, and federal regulatory mandates are key factors in making motor vehicles safer. Technologies developed in one regulatory context may reinforce other regulatory requirements. Mandates to reduce motor vehicle greenhouse gases, for example, are leading manufacturers to cut tailpipe emissions by using vehicle-to-vehicle communications that reduce unnecessary braking and acceleration, and enable more efficient driving patterns. Vehicle safety is enhanced by these changes. Similarly, the sensors and lidar that are being developed for automated vehicle safety may well help improve vehicle fuel efficiency. Reforming the Regulatory Process The development of a new Federal Motor Vehicle Safety Standard can be lengthy, often lasting many years. Former DOT Secretary Anthony Foxx and former NHTSA Administrator Mark Rosekind broadened the agency's approach beyond the traditional rulemaking to include new means of interacting with manufacturers and other vehicle safety stakeholders. In congressional testimony in November 2016, then-NHTSA Administrator Rosekind said the agency's regulatory process was too slow, given the pace of technological development. He explained that a traditional approach to regulating these new technologies would be to engage solely in rulemaking process, writing new regulations that prescribe specific standards. Our view is that approach would stymie innovation and stall the introduction of these technologies.... Any rule we might offer today would likely be woefully out-of-date by the time it took effect, given the pace of technological development.... Among the steps DOT and NHTSA took in 2016 to address these issues and establish new forms of enhancing vehicle safety are the following: Secretary Foxx announced a voluntary agreement in January 2016 with 18 automakers to collectively analyze and share safety data, increase the number of car owners who respond to recall notices, and develop a joint approach to automotive cybersecurity. While automakers supported the agreement, former NHTSA Administrator Joan Claybrook reportedly criticized it as ineffective. In March 2016, NHTSA and the IIHS announced a commitment of 20 vehicle manufacturers to make automatic emergency braking (AEB) a standard feature on virtually all new passenger vehicles by 2022. This voluntary agreement makes AEB standard on vehicles three years earlier than had NHTSA pursued a traditional rulemaking. In those three years, IIHS estimates that 28,000 crashes and 12,000 injuries will be prevented. NHTSA's September 2016 Federal Automated Vehicle s Policy officially adopted SAE International's levels of automation, and provides guidance to automakers and other vehicle developers with a 15-point "Safety Assessment" that discusses safety areas that manufacturers should evaluate in developing highly automated vehicles. In addition, the policy statement delineates federal and state roles in the absence of an FMVSS regulatory process for automated vehicles, and also discusses how NHTSA might use current regulatory tools—such as exemption and interpretation authorities—to expedite the development of safe highly automated vehicles. New Vehicle Safety Laws Congress dealt extensively with vehicle safety issues in the FAST Act, the five-year surface transportation law enacted in December 2015. Its provisions on vehicle safety are described below. Rental Cars82 Rental car companies with more than 35 vehicles must repair vehicles subject to recalls before renting, leasing, or selling them. NHTSA was given authority to investigate rental car company violations of recalls. Motor Vehicle Dealers84 Motor vehicle dealers are required to notify owners of open recalls when an owner brings a vehicle to the dealer for servicing. The provision does not require dealers in used motor vehicles to repair vehicles subject to a recall prior to selling them to consumers. Recall Notifications Several provisions address the low recall completion rate in many vehicle recalls and seek to boost vehicle owner participation in recall campaigns. In the past, the law required notification of consumers by first-class mail; the FAST Act expands the requirement to include electronic means of notification, including use of email, social media, and targeted online campaigns. DOT is required to conduct a series of multiyear analyses of recall completion rates and report the findings to Congress, including information on recall completion rates by manufacturer, model year, components, and vehicle type. NHTSA is also required to report on how it will improve recall completion rates based on the analyses. The DOT Inspector General is required to audit NHTSA management of safety recalls. The law also requires DOT to initiate a two-year pilot grant program with no more than six states to evaluate the feasibility of using each state's motor vehicle registration process to inform consumers of open recalls on their vehicles. In addition, DOT is given two years to adjust its website by using current information technology, web design trends, and other best practices to ensure that motor vehicle safety recall information is more easily accessible to the public. DOT is directed to study the feasibility of adding to each new vehicle a technical system that would tell the vehicle owner when the vehicle was subject to an open recall, and to report the findings to Congress within one year. This study has not been completed. Increase in Civil Penalties and Automotive Accountability For each violation of the law, the statutory civil penalty cap is increased from a maximum of $35 million to $105 million. It is thought that the risk of higher punitive penalties will encourage automakers to more readily disclose potential defects that could lead to a recall. In addition, the time period during which automakers must pay to remedy defects is increased from 10 to 15 years (after a consumer is notified of the recall); the time period they must retain safety records is doubled from five to 10 years. The law also includes a whistleblower provision that encourages industry employees to come forward with information about possible motor vehicle safety violations and allows DOT to pay awards to whistleblowers from a portion of recovered civil penalties. Driver Privacy92 The Driver Privacy Act of 2015 was included in the FAST Act, stipulating that data retained by an event data recorder (EDR) is the property of the vehicle owner. EDR data can be accessed by someone other than the owner only in certain circumstances, such as under a court order. Most vehicles include EDRs, and owner's manuals describe their use, but there was congressional concern over how this data could be used and who owns it. NHTSA is required to submit a report to Congress within one year, evaluating the amount of time EDRs should capture and record vehicle data that is sufficient to investigate the cause of motor vehicle crashes; and promulgate within two years a regulation establishing the appropriate time period for EDR data capture. Child Occupants Congress has shown concern about infants left in car seats for prolonged periods of time. A 2012 law recommended (but did not require) that DOT research methods to reduce these risks; the FAST Act requires DOT to initiate research into ways to reduce the risks of hyperthermia or hypothermia to children left unattended in vehicles' rear seats. It also requires NHTSA to revise its crash data collection system to capture additional information on types of child restraints employed in crashes and to report its findings to Congress. Crash Avoidance Disclosure97 DOT is required to develop a rulemaking that will add crash avoidance information, such as automatic braking and lane departure prevention, next to crashworthiness information on motor vehicle window stickers. Tires99 The FAST Act includes several provisions related to motor vehicle tires, including requirements that NHTSA update its standards for tire pressure monitoring, develop a rule for tire fuel efficiency minimum performance standards, and establish an electronically searchable tire recall database. The time period for remedying tire defects is extended from 60 to 180 days (from the time a consumer is notified of a recall). Issues Before Congress Although many of the changes in federal vehicle safety policy made by the FAST Act have yet to take full effect, Members of Congress have advanced several other proposals that would extend NHTSA's authority to regulate motor vehicles. Among them are: Obligations to Repair Recalled Vehicles Current law does not require auto dealers to fix used cars on their lots, or taxi and ride-sharing services to repair vehicles being used to transport customers. Some Members of Congress have called for including used cars in the recall process. When a House floor amendment was debated during consideration of the surface transportation bill in 2015, it was argued that auto dealers do not in practice sell cars with defects and that some recalls are "overly broad because the majority of vehicle recalls do not require the drastic step of grounding the vehicle." NHTSA presently has no authority to order repairs of recalled vehicles used by taxi and ride-sharing services. Imminent Hazard Authority Currently, NHTSA cannot require manufacturers to immediately stop sales of vehicles or equipment without following the substantial procedural steps needed to complete a recall investigation. The Obama Administration asked Congress to grant NHTSA "imminent hazard authority," which would allow the agency to take immediate action when it believed there was the likelihood of death or serious injury. Congress did not include such authority in the FAST Act. Public Access to Safety Information Some Members of Congress have called for amending the Early Warning reporting provisions to require NHTSA to make information it receives from manufacturers more publicly available in a searchable, website format, contending that consumers and safety analysts could better evaluate potential defects. Prohibition of Regional Recalls NHTSA may allow auto manufacturers to limit a recall to a certain geographic area if there is evidence that the defect is primarily found in vehicles registered in that area. For example, the recall of Takata airbags was initially deemed a regional recall because excess humidity seemed to play a role, so only vehicles in more humid parts of the country were subject to the recall. Critics contended that a regional recall was inappropriate because vehicles registered in other areas at the time of the recall could subsequently be sold or moved to high-humidity areas, putting owners and passengers at risk. The Takata recall was broadened to a national recall after airbag defects were found in vehicles in other parts of the country. Legislation proposed during the consideration of the FAST Act would have eliminated regional recalls. Cybersecurity The Security and Privacy in Your Car Act of 2017 ( S. 680 ) would direct NHTSA and the Federal Trade Commission to establish federal standards to secure connected features and other motor vehicle data from hackers and data trackers. The legislation would also require the two agencies to develop a "cyber dashboard" rating that would show on a vehicle window sticker how well each vehicle model protects security and privacy of vehicle owners. The Security and Privacy in Your Car Study Act of 2017 ( H.R. 701 ) would require NHTSA to report to Congress after conducting a study to determine the appropriate cybersecurity standards for motor vehicles, including how critical vehicle software systems can be separated from other software systems, and techniques necessary to prevent intrusions into motor vehicle software systems. Civil Penalties The Obama Administration asked Congress to increase the maximum civil penalty on a manufacturer for selling vehicles that violate Federal Motor Vehicle Safety Standards from $35 million to $300 million per violation. The FAST Act increased the maximum penalty to $105 million.

Summary: Federal motor vehicle safety regulation was established more than 50 years ago by the National Traffic and Motor Vehicle Safety Act (P.L. 89-563) to address the rising number of motor vehicle fatalities and injuries. The National Highway Traffic Safety Administration (NHTSA) administers vehicle safety laws and has issued dozens of safety standards, including regulations affecting windshield wipers, hood and door latches, tires, and airbags. NHTSA has estimated that between 1960 and 2012, federal motor vehicle safety standards saved more than 600,000 lives, and the risk of a fatality declined by 56%. Although dozens of technologies were made subject to federal standards in the decades after federal regulation began, a NHTSA study reported that more than half of the lives saved-329,000-were from use of seat belts. While the federal standard was helpful in reducing fatalities, the study found that the passage of state laws allowing police to issue tickets if a driver or passengers are not wearing seat belts caused the number of lives saved to climb from 800 per year to 6,000 per year. In addition to promulgating and enforcing vehicle safety standards, NHTSA investigates vehicle defects that affect safety and issues vehicle or parts recalls if safety defects are discovered. In recent years, the number of vehicle and parts recalls has risen significantly, from 16.3 million vehicles and parts in 2013 to 87.5 million in 2015. The rising number of recalls is due to stricter laws and reporting requirements, larger fines, delayed detection of vehicle problems by NHTSA, and several high-visibility cases, including General Motors' faulty ignition switch and Takata airbags. Recalls rarely obtain 100% completion rates, leaving many defective vehicles on the road long after a recall is initiated. A recent study by J.D. Power, a market research company, showed that between 2013 and 2015, recalls of fewer than 10,000 vehicles had a 67% completion rate, while recalls of more than a million vehicles had a completion rate of only 49%. The larger recalls are thought to result in fewer repaired vehicles because of the difficulty in finding and notifying larger numbers of owners, a lengthened repair period due to lack of an adequate supply of replacement parts, and the ability of manufacturers to use more personalized communications, such as telephone calls, in smaller recalls. Many emerging technologies, such as automatic emergency braking and lane departure warning, are expected to reduce vehicle injuries and deaths in the future. Over time, these separate technologies will be combined as vehicles are built with higher levels of automation. To deal with these rapid changes, NHTSA has broadened the agency's approach beyond the traditional rulemaking to include new means of interacting with manufacturers and other vehicle safety stakeholders, such as voluntary agreements to accelerate use of life-saving technologies. The 2015 Fixing America's Surface Transportation (FAST) Act included significant vehicle safety provisions, including a new requirement that rental car fleets be covered by recalls, new methods for notifying consumers about recalls, larger penalties for violations, and a longer period for consumers to obtain remedies for defects. Congress remains interested in motor vehicle safety; proposed legislation calls for used vehicles to be subject to recalls, NHTSA to provide more public access to safety information, civil penalties to be increased, regional recalls to be terminated, and federal standards to be issued to secure electronic motor vehicle data from hackers.

gov_report

en

Summarize: NEW YORK (CNNMoney) Best Buy CEO Hubert Joly has led a turnaround at the company that's sent shares soaring. And that's turned into a very big payday for his ex-wife. On Tuesday Joly disclosed in a filing that he sold 451,153 shares of the company for a total of $16.7 million. He paid $6.3 million to exercise stock options, so he netted just over $10 million through the sale. The company issued a statement saying the sale was prompted by his need to pay a divorce settlement. "This sale reflects only one thing -- Mr. Joly has recently gone through a divorce and needs to sell a portion of his holdings in order to cover the costs of that unfortunate event," said the company's statement. "He remains heavily invested in Best Buy." Related: Best Buy - not your standard corporate comeback The company says Joly's sale represented about 20% of his Best Buy (BBY) stake. Tuesday's filing shows he holds about 476,000 shares after the sale, but a spokesman said Joly has received additional stock grants and options that have not yet been disclosed in company filings. Joly was born in France and previously worked at Carlson Wagonlit Travel, home of such brands as Radisson and TGI Friday, before he was named CEO of Best Buy on Aug. 20, 2012. Related: Rebuild your nest egg after a divorce The company has struggled with weak sales and growing competition from online retailers such as Amazon (AMZN). Shares continued to decline in his first few months on the job, especially as founder and former CEO Richard Schulze failed at his effort to take the company private. But shares have tripled in value so far this year as Joly cut costs and improved earnings, making Best Buy one of the top performers on the S&P 500 index. Dow Jones Reprints: This copy is for your personal, non-commercial use only. To order presentation-ready copies for distribution to your colleagues, clients or customers, use the Order Reprints tool at the bottom of any article or visit www.djreprints.com

Summary: Best Buy had to issue an unusual statement explaining exactly why its CEO just dumped 20% of his stake in the company: the business his fine; his marriage, not so much. Hubert Joly sold about 450,000 shares and netted $10 million, reports CNNMoney. "This sale reflects only one thing-Mr. Joly has recently gone through a divorce and needs to sell a portion of his holdings in order to cover the costs of that unfortunate event," said the company statement. A little awkward, sure, but better than the mess surrounding the previous CEO, who resigned over allegations of improper conduct with a female employee, notes the Wall Street Journal.

multi_news

en

Write a title and summarize: L locus resistance (R) proteins are nucleotide binding (NB-ARC) leucine-rich repeat (LRR) proteins from flax (Linum usitatissimum) that provide race-specific resistance to the causal agent of flax rust disease, Melampsora lini. L5 and L6 are two alleles of the L locus that directly recognize variants of the fungal effector AvrL567. In this study, we have investigated the molecular details of this recognition by site-directed mutagenesis of AvrL567 and construction of chimeric L proteins. Single, double and triple mutations of polymorphic residues in a variety of AvrL567 variants showed additive effects on recognition strength, suggesting that multiple contact points are involved in recognition. Domain-swap experiments between L5 and L6 show that specificity differences are determined by their corresponding LRR regions. Most positively selected amino acid sites occur in the N- and C-terminal LRR units, and polymorphisms in the first seven and last four LRR units contribute to recognition specificity of L5 and L6 respectively. This further confirms that multiple, additive contact points occur between AvrL567 variants and either L5 or L6. However, we also observed that recognition of AvrL567 is affected by co-operative polymorphisms between both adjacent and distant domains of the R protein, including the TIR, ARC and LRR domains, implying that these residues are involved in intramolecular interactions to optimize detection of the pathogen and defense signal activation. We suggest a model where Avr ligand interaction directly competes with intramolecular interactions to cause activation of the R protein. The plant immune system is based upon the ability to accurately perceive and appropriately respond to potential threats. In general, plants use membrane-spanning proteins with extracellular receptor domains to recognize common features of plant pathogens (pathogen associated molecular patterns, PAMPs) and intracellular receptors to detect pathogen effectors transferred into plant cells during infection [1], [2], [3]. Most intracellular immune receptors (disease resistance proteins) contain nucleotide-binding (NB) and leucine-rich repeat (LRR) domains; one subclass of these has a coiled-coil (CC) domain and the other possesses a TIR (Toll, interleukin-1 receptor, resistance protein) domain at the N-terminus [4], [5]. Plant NB-LRR disease resistance proteins belong to the STAND (signal transduction ATPases with numerous domains) clade of AAA+ (ATPase associated with diverse cellular activities) proteins, and are similar to the Nod-like receptor (NLR) family of proteins that act as intracellular surveillance molecules in animal innate immunity [6], [7], [8]. The signature catalytic core of STAND proteins comprises an αβα NB domain, a four-helix ARC1 (APAF-1, R protein, CED-4) domain, and a winged helical ARC2 domain [9], [10], [11]. This domain is thought to function as a reversible molecular switch during signal transduction, with monomeric ADP-bound forms representing the off - or closed - state, and ATP-bound multimeric forms representing the on - or open – state [9], [11], [12], [13]. Tight regulation of this switch is critical in plant NB-LRRs, because these proteins regulate an apoptotic process. The trigger for the conformational change to the open state is generated by signal perception, either directly when NB-LRRs bind effector proteins [14], [15], [16], [17], [18], [19], or indirectly when NB-LRRs detect the biochemical fingerprint of effector proteins as they attempt to carry out their virulence function [20], [21], [22], [23], [24], [25]. This effector-mediated R protein activation is believed to ultimately lead to conformation changes that expose the N-terminal TIR or CC signalling domains, so they can interact with downstream signalling partner [12], [26]. The C-terminal LRR domain of R proteins generally mediates signal perception [27], [28]. This domain is composed of repeating LRR units that form stacking β-strands, resulting in a horseshoe-shape molecule with a continuous, parallel β-sheet on the inner concave surface [29]. Individual LRR units contain xxLxLxx motifs generating β-strand/β-turn structures in which the variable non-leucine residues form the concave, solvent-exposed surface of the horseshoe and are available for participation in protein-protein interactions [29], [30]. This region of plant R proteins is often highly variable, as a result of diversifying selection, and a number of studies have demonstrated changes in specificity mediated by polymorphisms in the LRR domain [18], [30], [31], [32], [33], [34], [35], [36], [37], [38]. TIR-NB-LRR resistance proteins in flax (Linum usitatissimum) confer resistance to the flax rust fungus Melampsora lini through recognition of effector proteins delivered into the host cell during infection [39], [40]. For example, the L resistance locus consists of a single gene encoding 13 allelic protein variants (L, L1 to L11, and LH) that recognise different matching avirulence proteins [32]. L alleles share greater than 90% amino acid sequence identity, with positively selected variation concentrated in the LRR domain. Domain-swap experiments between the L2, L6 and L10 alleles showed that these recognition specificities are determined by the LRR domain [30], [32]. Similarly, the L6 and L11 proteins differ by only 32 amino acids, all in the LRR domain, and a chimeric protein with 11 amino acid changes in the C-terminal region of the LRR displayed a novel specificity, with a reduced recognition spectrum [16], [41]. The L5, L6 and L7 proteins recognise allelic variants of the M. lini effector protein AvrL567, a 127-amino acid secreted protein that is expressed in haustoria and translocated into host cells during infection [42], [43], [44]. Seven of the 12 variant forms of AvrL567 (-A, -B, -D, -E, -F, -J, -L) are avirulence alleles as they induce an L5 and/or L6, and/or L7-dependent hypersensitive response (HR) in transient expression assays whereas the other 5 variants (-C, -G, -H, -I, -K) are virulence alleles as they do not induce an HR [16]. Yeast-two-hybrid (Y2H) assays demonstrated that AvrL567 and L5, L6, and L7 interact directly and that the specificity of this protein-protein recognition corresponds with that of the HR-inducing recognition in planta [16]. L6 and L7 are differentiated by just 11 polymorphisms found in the TIR domain and have identical AvrL567 recognition specificities, although L7 shows consistently weaker interaction in yeast, and a weaker HR in planta [26], [30]. L5 and L6 are two of the most diverged L proteins, differing by 89 amino acid polymorphisms (61 in the LRR) and four small indels, but nevertheless have overlapping recognition specificities. They are distinguished by L6 interacting with AvrL567-D, while L5 does not. Wang et al. [45] determined the structures of AvrL567-A and -D and identified four polymorphic surface-exposed amino acid residues that were important for their differential recognition. Here we have further investigated the role of these surface-exposed amino acids in recognition of AvrL567. Single, double and triple mutations at these sites in a variety of AvrL567 variants showed additive effects on recognition strength, suggesting that multiple contact points are involved in the recognition event. We show by domain-swap experiments that the L5 and L6 specificities are determined by their corresponding LRR regions, with contributions made by seven and four N- and C-terminal LRR units, respectively, of a total of 26, where most positively selected amino acid sites occur. This further confirms that multiple, additive contact points occur between AvrL567 variants and either L5 or L6. However we also observed that recognition of AvrL567 is affected by co-operative polymorphisms between both adjacent and distant domains, including the TIR, ARC and LRR domains, implying that these residues are involved in intramolecular interactions to optimize detection of the pathogen and/or defense signal activation. Sequence comparisons of the 12 AvrL567 variants suggested that polymorphisms at four positions (50,56,90 and 96) were associated with specificity differences [16]. Single amino acid substitutions at positions 50 (T50I) or 96 (L96R) were sufficient to restore recognition of AvrL567-D by L5, while the I50T substitution almost completely blocked L5 and L6 recognition of AvrL567-A (Table 1) [45]. To further evaluate the role of these residues in mediating interactions with L5 and L6, we made reciprocal single, double and triple substitutions of these amino acids in a wider range of AvrL567 variants (-A, -D, -E, -J and -C), which show varying recognition patterns (Table 1). Mutant AvrL567 proteins were assayed for recognition by L5 and L6 using both Y2H assays - to test for protein interaction - and by Agrobacterium-mediated transient expression in planta - to measure R gene-dependent cell death (Figure 1 and Table 1). With one exception (see below), single amino acid changes at positions 56,90 and 96 in AvrL567-A did not alter recognition by L5 or L6 [45]. However, double and triple substitutions at these positions revealed that they all contribute additively to recognition. The K56D/S90I and K56D/R96L double mutants both substantially reduced recognition by L5, while the K56D/S90I/R96L triple substitution blocked L5 recognition completely (Figure 1). Notably, none of these changes affected L6 recognition. Conversely, the single amino acid R96S substitution abolished L6 but not L5 recognition. This indicates that L5 and L6 recognise different molecular features of AvrL567, but at similar positions. For AvrL567-D, Wang et al. [45] showed that either T50I or L96R substitutions were sufficient to allow interaction with L5; but we now found that the T50I/L96R double mutation shows an additive effect relative to the single mutants, which can be detected when the GAL4 AD and BD fusions are reversed (Figure S1). Further evidence for additive interactions comes from the context-dependent effects of several single substitutions. For instance, the presence of S or L at position 96 does not prevent L5 recognition of AvrL567-A or -J, but in AvrL567-D an R is required at this position to establish L5 recognition. Likewise, a S96 substitution destabilizes L6 recognition of both -A and -D, but is compatible with L6 recognition of -J (Figure 1 and Table 1). To complement these loss-of-function studies, we also tested the effect of reciprocal changes in the virulence allele, AvrL567-C, which is not recognised by L5 or L6 and found that multiple amino acid changes were required to restore full recognition (Figure 1, Table 1). For instance, double substitutions at positions 50 and 56 or positions 50 and 96 were required to allow L5 recognition of this protein. L6 recognition could be restored weakly (in yeast but not in planta) by the single S96R substitution, but required the triple T50I/D56N/S96R substitution for full recognition. Interestingly, in the context of AvrL567-C, a K residue at position 56 was not compatible with L6 recognition, although it does not prevent recognition in the AvrL567-A or -D contexts (Table 1). The strong positive effect of isoleucine at position 50 was confirmed as the single T50I substitution in AvrL567-E restored its interaction with L5 and L6. These data further support the additive roles of these amino acid positions in recognition. All AvrL567 mutant fusion proteins were stably expressed in yeast (Figure S2) indicating differential recognition of mutants by L proteins was due to differences in their surface properties, resulting in physical changes in the interactions of these proteins. Data from Y2H and in planta HR analyses correlated well, apart from a few exceptions, which can all be explained by in planta HR induction being less sensitive than the Y2H interaction (for instance, L6 interactions with AvrL567-A K56N or AvrL567-J S96L; Figure 1). Overall, these data suggest that multiple contact points at disparate positions on the AvrL567 molecule are involved in interaction with the corresponding R proteins and make additive contributions to the strength of recognition. In addition, although L5 and L6 recognition of AvrL567 involves contacts with similar positions, they have different requirements for amino acid residue features at these positions. The cloned Avr genes and their derived mutants provide a sensitive set of test proteins to detect subtle changes of specificity of L5–L6 chimeric proteins described in the following sections. In order to correlate the recognition-determining residues in the AvrL567 proteins with variation in the L5 and L6 proteins, we conducted an analysis of positive selection on the coding sequences of all 12 cloned L genes. Previous analysis had found an excess of non-synonymous versus synonymous substitutions in the LRR domain [30], and we used the program CODEML [46] to identify codons under positive selection. The M8 model allowing for positive selection provided a significantly better fit to the data than the M7 null hypothesis model, which excludes positive selection (p<0. 001; Table S1) and predicted 123 (9. 5%) codons as being under significant positive selection (Figure 2). This includes 86 of the 99 sites polymorphic between L5 and L6 (Figure S3). To examine the distribution of positively selected sites we considered the protein sequence in six regions: the TIR, NB, ARC1, ARC2 and LRR domains and a short spacer region between ARC2 and LRR domains. In the LRR domain, 13. 2% of codons (92 sites) are under significant positive selection, compared to only 5. 3% of codons (31 sites) in the rest of the protein. These occurred mainly in the N-terminal and C-terminal portions of the LRR domain, with a lack of positively selected sites in the central portion of the LRR domain. This suggests that recognition specificity may be conferred mainly by interactions involving the two extremities of the LRR domain. The ARC1 domain and the spacer also showed elevated numbers of positively selected sites (about 11%) compared to the TIR, NB and ARC2 domains (2 to 5%; Figure 2). The concentration of positively selected sites in the LRR domain of L proteins is consistent with the proposed role of this domain in recognition specificity. To test whether polymorphisms in the LRR domains of L5 and L6 are responsible for their different recognition specificity, we firstly generated chimeric proteins L6592L5 and L5592L6, in which the complete LRR domains of L5 and L6 are exchanged at an engineered AvrII restriction site in codons 592–593 (Figure S4). The introduction of this site causes a W to R amino acid change at position 592, but this substitution did not effect AvrL567-A or -D recognition by the modified L5 or L6 alleles (Figure S5) and is also found in the functional L9 protein. The chimeric proteins were tested for recognition of AvrL567 variants and mutants by the Y2H assay (Figure 3). Both recombinant proteins were well-expressed in yeast (Figure S2), but L5592L6 was non-functional in that it did not interact with either AvrL567-A or -D (Figure 5a). This may be related to the position of the exchange site within a seven-amino acid indel polymorphism (Figure S3). On the other hand, L6592L5 exhibited L5-like specificity, giving recognition of AvrL567-A but not -D, which indicated that this recognition pattern was determined by the LRR domain of L5 (Figure 3b). However, when tested against the extended set of AvrL567 mutants, L6592L5 recognized only a subset of the wild-type L5 repertoire, and failed to interact with AvrL567-A K56D, K56D/S90I and K56D/R96L mutants, and with most AvrL567-C gain-of function mutants (Figure 3b). We therefore extended the region swapped between the alleles further towards the N-terminus. The chimera L6493L5, which contained the L5 LRR domain plus an additional three amino acid polymorphisms from the ARC2 domain and the entire wild-type spacer region (Figure 3a, S3), retained the full L5 recognition specificity across the pool of AvrL567 variants (Figure 3b). The only exceptions were a slightly enhanced interaction with the AvrL567-A I50T and K56D/S90I mutants, which only weakly interacted with L5, and a weak interaction with AvrL567-D L96S, which did not interact with L5. Similarly, the chimera L5556L6 that included the L6 LRR domain and the spacer region had L6-like recognition specificity when tested against wild-type AvrL567 variants and mutants (Figure 3c, d), although in some cases interactions were weaker than for L6 (AvrL567-D, AvrL567-D N56K and L96R and the AvrL567-C mutants). In conclusion, the data are consistent with the L5–L6 specificity differences being contributed by polymorphisms in the LRR domain and spacer region, but with the strength of the R: Avr protein interactions modulated somewhat by interactions between these regions and the N-terminal TIR-NB-ARC region. The recognition repertoires conferred by the LRR domains of L5 and L6 can be qualitatively distinguished by their interactions with AvrL567-D (interacts with L6 but not L5) and the AvrL567-A mutant R96S (interacts with L5 but not L6) (Figure 1 and 3). To further understand the role of LRR domain polymorphisms in these differences, a series of L5–L6 chimeras with swaps within the LRR domain was generated. We designed swaps that would exchange groups of positively selected amino acids, as well as residues implicated in interaction in the docking-derived models of AvrL567 binding to a modelled L5 LRR structure presented by Wang et al. [45] (Figure S3). All the chimeric proteins were stably expressed in yeast (Figure S2) and were evaluated for interactions with AvrL567-A, -D and the AvrL567-A-R96S mutant (Figure 4). These experiments allowed us to draw several inferences about residues controlling recognition specificity. Firstly, interaction with AvrL567-D, which discriminates the L6 specificity, requires L6 polymorphisms in the last four LRR units. Substitution of these four LRR units of L6 with the corresponding region of L5 (L61193L5), abolished the interaction with AvrL567-D, but not -A (Figure 4a). This was also true for the same LRR exchange made in the context of the L5 TIR-NB-ARC (L5556L61193L5; Figure 4b). Conversely, the reciprocal exchange in L5 (L51193L6) allowed weak interaction with AvrL567-D (Figure 4c), suggesting that the L6 polymorphisms in the last four LRR units are both necessary and sufficient to confer AvrL567-D interaction in these proteins. Interestingly, this interaction was stronger when the L6 TIR-NB-ARC region was also present (L6592L51193L6; Figure 4d). Indeed, the L6 TIR-NB-ARC region enhanced the recognition of AvrL567-D for all the chimeras containing the L6 C-terminal LRRs (compare Figures 4c and 4d). Secondly, the ability to interact with AvrL567-A-R96S, which discriminates the L5 specificity, requires L5-specific polymorphisms in the first seven LRR units. L6592L5, containing the full-length L5 LRR domain, interacted with AvrL567-A-R96S (Figure 3), while proteins with chimeric L6-L5 LRR domains containing L6 N-terminal LRR polymorphisms (L6793L5, L6972L5, L61125L5 and L61193L5) did not (Figure 4a). Conversely, chimeras containing the reciprocal region from L5 swapped into the remainder of L6 (L6592L5793L6, L6592L5972L6, L6592L51125L6, and L6592L51193L6) acquired the capacity to interact with AvrL567-A-R96S (Figure 4d). Thus, the L5 polymorphisms found between residues 594 and 793 in L5 are necessary and sufficient to determine the AvrL567-A-R96S interaction in these proteins. Intriguingly, two chimeric proteins (L5793L6 and L5972L6), which contain the critical 594-to-793 L5 residues along with the L5 TIR-NB-ARC domains did not interact with AvrL567-A-R96S (Figure 4c). As above, this suggests that the presence of L6 TIR-NB-ARC region is required for strong Avr protein interactions in proteins containing the L6 C-terminal LRRs. Neither L6493L5972L6 nor L6493L51125L6 interacted with AvrL567-D (Figure 4e), suggesting an additional positive contribution to the strength of the interaction between these LRR chimeras and AvrL567-D may be attributed to the presence of one or more of the three L6-specific ARC2 polymorphisms (Figure S3). We also observed that certain L6-L5 LRR chimeras lacked recognition function. For instance, swaps containing the N-terminal LRRs of L6 and the C-terminal LRRs of L5 gave rise to non-functional proteins when the junctions were made at positions 793 or 1125, but not at 972 or 1193 (Figure 4a and 4b). Similarly, proteins containing the N-terminal LRRs of L5 and the C-terminal LRRs of L6 gave rise to proteins with reduced functionality when the junctions were made at positions 793,972 or 1125 (Figure 4c), although this could be overcome by the presence of polymorphisms found in the L6 TIR-NB-ARC region (compare swaps in Figure 4c, d and e). These observations suggest a requirement for specific, co-operative combinations of polymorphisms within the LRR domain to allow interaction with the corresponding ligand, consistent with the interaction occurring across a large surface area. Because the strength of AvrL567 interaction of several chimeric proteins appeared to be influenced by whether the TIR-NB-ARC region is derived from L5 or L6, we decided to examine the influence of polymorphisms in the N-terminal region on Avr protein interaction. A series of chimeras were generated in which various regions of the L6 TIR, NB, ARC1 and ARC2 domains were re-introduced into the L5556L6 protein, which exhibited L6-like specificity, but weaker AvrL567-D interaction, and tested for interaction with AvrL567-A and -D (Figure 5a). All chimeric proteins were stably expressed in yeast (Figure S2). Introduction of increasing lengths of L6 sequence from the N-terminus (L6185L5556L6 to L6431L5556L6), including the TIR and NB regions, did not increase the interaction with AvrL567-D, but a further swap including the L6 ARC1 (L6493L5556L6) restored strong interaction with AvrL567-D. This suggested that one or more of the six amino acid polymorphisms between 447 and 484 (five in ARC1 and one in ARC2) were responsible for the reduced interaction. Consistent with this, inclusion of the ARC1 and ARC2 regions of L6 along with the L5 TIR-NB (L5431L6) also restored recognition of AvrL567-D, again implicating this region in the reduced interaction. Chimeric proteins representing the inverse swaps and including the L5 LRR domain (L6414L5, L6431L5, L6493L5, L6592L5, L5185L6592L5 and L5226L6592L5) did not interact with AvrL567-D but retained interaction with AvrL567-A (Figure 5b), similar to L5. This suggests that polymorphisms in the LRR domain of L6 are required to provide the specific recognition capacity to bind to AvrL567-D, while those in the ARC1/2 region may contribute to the strength of the interaction through intramolecular interactions. Interestingly, some other swaps in the TIR-NB region also led to reduced Avr protein interaction. Notably, while the L5185L6 chimera was functional, L5226L6 did not interact with either AvrL567-A or -D. Similarly, L6185L5556L6 failed to interact with the Avr proteins, while L6226L5556L6 did interact with AvrL567-A. However, both the reciprocal swaps (L5185L6592L5 and L5226L6592L5, Figure 5b) interacted with AvrL567-A. This suggests that the two L5-derived amino acid polymorphisms in this region (E216 and L218) interfere with recognition in the context of the L6 LRR domain. We therefore tested a series of constructs containing chimeric L5–L6 LRR domains in the context of the L5226L6 protein, to determine which part of the L6 LRR domain mediates this incompatibility (Figure 6). Introduction of the seven N-terminal LRR units from L5 was sufficient to restore AvrL567 interaction in this protein (Figure 6a), while all chimeras containing this region from L6 failed to interact (Figure 6b). A similar pattern was observed for chimeras containing a hybrid L6185L5 TIR-NB junction (L6185L5556L6, L6185L5556L6592L5793L6 and L6185L5556L6592L51193L6; Figure 5a and Figure 6c). This suggests that a negative interaction occurs between these TIR domain polymorphisms of L5 and polymorphic residues in the N-terminal region of the L6 LRR. Even though several of these constructs contained all of the polymorphisms from L6 that are required for strong recognition of AvrL567-D, the presence of the hybrid L5–L6 TIR-NB junction appears to have prevented this interaction, for example in L6185L5556L6592L5793L6 and L6185L5556L6592L51193L6 (Figure 6c). As described above, chimeras L6493L51193L6 and L6592L51193L6, which contain the N-terminal L5 specificity region and the C-terminal L6-specificity region, represent a novel and expanded recognition specificity, in that they recognise both AvrL567-A-R96S and AvrL567-D, which distinguish L5 and L6 (Figure 4). We therefore tested these further against the larger set of AvrL567 variants and mutants and compared this to the recognition repertoires of L5, L6, L6493L5 and L6592L5 (Table 2 and Figure S6). The L6592L51193L6 and L6493L51193L6 recognition specificities were largely L6592L5-like and L6493L5-like, respectively, however both were L6-like in regards to interactions with AvrL567-D and its derived mutants. Overall, L6493L51193L6 recognized more AvrL567 variants and mutants (23) than L6 (21), L6493L5 (20), L5 (19), L6592L51193L6 (14), or L6592L5 (13) (Table 2). We further tested the L6493L51193L6 chimera for its ability to trigger an AvrL567-dependent cell death in planta (Figure 7). Agrobacterium-mediated transient expression of the L5, L6 and L6493L51193L6 cDNAs in transgenic tobacco also expressing AvrL567-A induced a strong HR-like cell death response (Figure 7a), confirming that the L6493L51193L6 construct expresses a functional resistance protein. Co-expression of L6493L51193L6 with AvrL567-D or AvrL567-A-R96S also induced an HR, while L5 induced cell death only with AvrL567-A-R96, and L6 only with AvrL567-D (Figure 7b, c), thus recapitulating the recognition specificity observed in yeast. No HR was observed when L6493L51193L6 was expressed alone (Figure 7d), indicating that this chimera is not autoactive. Mutational analysis shows that multiple sites are involved in the interaction between AvrL567 variants and L5 and L6, and that L5 and L6 have different specificity requirements at these positions (Figure 1). The additive nature of the interactions is shown by the effects of double and triple mutations in the AvrL567 proteins on recognition. For instance, while single mutations at positions 56,90 and 96 do not disrupt recognition by L5, triple substitution abolishes L5 recognition. However, these changes do not disrupt recognition by L6, highlighting the different sequence requirements of the two resistance proteins. Conversely, with the exception AvrL567-C-S96R (which is weakly recognised by L6), the virulence allele AvrL567-C required at least two to three mutations in combination to allow full recognition by L5 or L6. Again, the requirements for the two resistance proteins were different. A double substitution at positions 50 and 56 was sufficient for L5 recognition, while full L6 recognition required a further substitution at position 96, and showed a requirement for an asparagines residue at position 56 rather than lysine. Previously, Wang et al. [45] demonstrated that T50 in AvrL567 destabilizes interactions with L5 and L6 and our data further confirm that this residue is particularly important for recognition. For instance, the T50I substitution has a strong positive effect in AvrL567-E on stabilizing interactions with L5 and L6 (Figure 1). AvrL567-E and -J differ by only two polymorphisms (H26D and T50I), but we previously found that the N-terminal region consisting of amino acids 26–37 of AvrL567-A could be deleted without affecting recognition [43]. Therefore, the T50I polymorphism is the critical residue that differentiates AvrL567-E and -J recognition specificities. The importance of position 50 can also be observed by the contribution a T50I substitution makes to allow L5 recognition of AvrL567-C when paired with D56N, D56K, or S96R substitutions, and to allow L6 to recognize AvrL567-C when associated with D56N and S96R in a triple substitution (Figure 1). The presence of a D residue at position 56 had a small negative effect on interactions with L5 but not L6 [45], and we observed that this effect is much stronger when combined with either, or both, S90I and R96L in AvrL567-A. Similarly, neither S90I or R96L substitutions (Table 1), nor the double S90I/R96L substitution in AvrL567-A compromise recognition by L5; however, I90 and L96 both have stronger negative effects on interactions with L5 when combined with D56, and completely disrupt L5 recognition when all three substitutions are present in AvrL567-A (Figure 1). Interestingly, in the context of AvrL567-C, K56 has a negative effect on recognition by L6 but not L5, as the triple T50I/D56N/S96R substitution in AvrL567-C is recognized by both L5 and L6 in yeast, whereas the triple T50I/D56K/S96R substitution in AvrL567-C is only recognized by L5. Position 96 in AvrL567 is important for interactions with both L5 and L6, with R at this position favouring interactions with L5, and an S disrupting interactions with L6 (Figure 1 and [45]). For instance, the R96S substitution in AvrL567-A destabilizes interactions with L6, whereas the reciprocal S96R substitution improves interaction of AvrL567-C with L6 both individually, and in combination with T50I or T50I/D56N substitutions, and with L5 when combined with the T50I, T50I/D56N or T50I/D56K substitutions. However, as with other polymorphisms in AvrL567, the disruptive effect of S96 on L6 recognition is context-dependent, as both AvrL567-E and –J contain this polymorphism while maintaining interactions with L6. Collectively, these data indicate that L5 and L6 interact with AvrL567 through multiple amino acid contact points, and support the hypothesis that recognition is mediated in an additive manner by the cumulative composition and context of their amino acid sequences. Chimeric L5–L6 proteins containing reciprocally swapped LRR domains showed AvrL567 interaction specificities consistent with the origin of the LRR domain (Figure 3), although some interactions were weaker than in the wild-type proteins. Positively selected amino acid sites in L alleles cluster at the N and C-terminal regions of the LRR (Figure 2), and docking analysis of AvrL567 to the modelled L5 LRR domain [45] suggested that the most likely binding site of AvrL567 is between the two ends of the LRR with most potential contact points in these N- and C-terminal regions. This hypothesis is supported by analysis of LRR chimeras (Figure 4), which showed that 13 polymorphisms in the last four LRRs of L6 are required for the recognition of AvrL567-D, while polymorphisms in the first seven LRRs of L5 are required for recognition of AvrL567-A-R96S. Some internal LRR fusions (eg. L6793L5 and L61125L5; Figure 4a) fail to interact with AvrL567 variants, or show a reduced interaction repertoire (eg. L6493L5972L6, L6493L51125L6; Figure 4e), suggesting that surfaces involved in specific interactions may have been disrupted by fusions at these junctions. Previously, Ellis et al. [41] showed that polymorphisms between L6 and L11 in the last three LRRs (24,25 and 26) are important for L6 recognition of AvrL567. The L6L11RV chimera differs from L6 by 11 polymorphisms in these three LRR units, and recognizes only AvrL567-J, whereas L6L11B2, with two additional polymorphisms in LRR 23, does not interact with any AvrL567 variant (Figure S7). L5 is quite similar in sequence to L11 in this region, but a similar L6-L5 exchange in this region (L61193L5) maintained interaction with many AvrL567 variants, including -A and -J (Figure 4). Comparison of the C-terminal sequences of these chimeras (Figure S7) narrows the L11 polymorphisms responsible for these difference down to R1220 and K1222 in LRR 24 – contributing to the loss of Avr recognition (other than -J) in L6L11RV, and V1196 leading to the loss of -J recognition in L6L11B2, because these are the only polymorphisms unique to L11. It is important to note that these domain-swap experiments only examine the roles of polymorphic residues in determining recognition specificity and do not address the role of shared residues in AvrL567 interaction. Indeed, docking analysis identified 31 residues common to L5 and L6 that may be involved in protein contacts. Because chimeras with the N-terminal LRRs of L5 and C-terminal LRRs of L6 can interact with AvrL567-A, -A-R96S and -D, and reciprocal chimeras also retain interaction with at least AvrL567-A (Figure 4), we conclude that binding of AvrL567 to L5 and L6 occurs in the same basic orientation. The ability of the L5 N-terminal LRRs to allow binding to AvrL567-A-R96S may suggest that the AvrL567 surface region containing this residue makes contact with the N-terminal LRR region. Although in general domain-swaps involving the full LRR domain showed the expected specificity for the source of the LRR, some chimeras had weak or no interactions with AvrL567 variants (Figure 3). Because all of the polymorphisms found in L5 and L6 are compatible with AvrL567 interaction in their native context, this suggests that certain polymorphisms in L5 and L6 N-terminal regions occur in specific, co-operative combinations that are required for recognition function. We observed such co-adaptation between the spacer region and the LRR domain, between the ARC domains and the C-terminal region of the LRR domain, and between the TIR and LRR domains. While both L6493L5 and L6592L5 exhibited L5-like specificity (Figure 3), when tested on the wider set of AvrL567 mutants, L6592L5 recognised only a subset of the wild-type L5 repertoire. This suggests that some or all of the nine L5-specific amino acids in the ARC2 and spacer region (Figure S3) are important for optimal recognition. Six of these residues represent a small insertional polymorphism in the spacer region, which may influence the relative positioning of the L5 LRR domain with respect to the rest of the protein. A wild-type spacer region is probably also required for L6 to adopt a functionally competent state, because an exchange within this indel in L5592L6 resulted in a non-functional protein, while L5556L6 (in which the exchange occurs before this indel) showed an L6-like recognition repertoire (Figure 3). Given its proximity to the LRR domain, it is possible that the spacer region also participates directly in AvrL567 binding. On the other hand, the observed co-adaptation between polymorphisms at the C-terminus of the TIR domain and the N-terminus of the LRR domain, and between the ARC and LRR domains, likely reflects an indirect effect on ligand affinity as a result of intramolecular interactions that obscure the ligand-binding site, rather than a direct effect on binding specificity. Such general effects on ligand accessibility would be expected to manifest themselves particularly in the case of those interactions that are close to the threshold of detection in Y2H assays. Sequence exchanges involving regions of the TIR domain suggested that two L5-derived amino acid polymorphisms in this region (E216 and L218) interfere with recognition in the context of the L6 LRR. However, the TIR domain is not required for L6-AvrL567 interaction, and these two residues are exposed on the surface of the TIR domain structure in a region implicated in negative regulation of L6 through intramolecular interactions [26]. Indeed, TIR domain residues that are polymorphic between L6 and L7 also play a role in AvrL567 interaction and are responsible for the weak resistance phenotype of L7 [26], [30]. Likewise, polymorphisms in the ARC1 and ARC2 domains of L6 strengthen AvrL567-D recognition, conferred by the polymorphisms found in the last four LRRs of L6 (Figure 5). Again, this appears to be a general ligand affinity effect, because swapping the L6 ARC domains into L5 does not generate recognition of AvrL567-D (Figure 5b). Furthermore, the presence of the L6 TIR-NB-ARC region also strengthens interactions with AvrL567-A-R96S, mediated by polymorphisms found in the first seven LRRs of L5 (Figure 4c and d). We previously showed that a P-loop mutation (K271M) in L6, which would prevent nucleotide binding, also disrupted interaction with AvrL567 [16], consistent with the idea that interactions between the NB-ARC and LRR are required to support ligand binding. Previous experiments in other systems have also demonstrated intramolecular interactions between R protein domains that are important for function. The CC, NB-ARC and LRR domains of potato Rx can interact and functionally complement each other when expressed as separate polypeptides [47]. Likewise, domain swaps have also implicated co-adaptation between domains of Rx, Mi-1. 2 and I-2 in tomato, and Pm3 in wheat [34], [48], [49], [50]. It has been suggested that ARC1 functions as a molecular scaffold, forming intramolecular interactions with the LRR domain, and that signal perception disrupts these interactions [34]. Subsequently, ARC2 may transduce this effect into defence protein activation [50]. However, these experiments do not distinguish between effects on ligand recognition, protein activation or downstream signalling. Our data on AvrL567 binding by L5/L6 recombinants indicate that these intramolecular interactions can have direct effects on ligand binding. This suggests a model of R protein activation in which ligand binding occurs in direct competition with intramolecular interactions, which presumably maintain the resting protein in an inactive signalling state. Rather than Avr binding directly destabilising intramolecular interactions, it is possible that R proteins exist in an equilibrium between active and inactive states, with the Avr protein preferentially binding to and stabilising the active state to induce signalling. This competition provides a mechanism for signalling activation, as well as for fine-tuning the triggering of the response. Weak R: Avr interactions require a very delicate trigger if they are to induce effective resistance, but this would come at a cost of increased autoactivity of the R protein. Conversely, stronger Avr interactions could compete with more stable inhibitory intramolecular interactions. To visualize structurally the L protein regions involved in AvrL567 recognition, we prepared a homology model of the L6 NB-ARC domain with the program Modeller [51] using the multiple sequence alignment of NB-ARC domains from different R proteins published in van Ooijen et al. [11] and the crystal structure of APAF-1 as a template [52]. In the model, four of the L6 polymorphisms involved in strengthening interactions with AvrL567-D (A454, E457, E461, and R465) map to a solvent exposed region in the last α-helix of the ARC1 domain (Figure 8). This α-helix (H8 of HD1), is part of the ARC1-ARC2 linker region, found in APAF-1 and CED-4, that undergoes a drastic change during the switch from closed to open states [9]. Therefore in L6, these residues are positioned appropriately to be involved in either, or both, inter- and intramolecular interactions, and may provide a putative link between effector perception and hypothetical conformational changes that lead to the open state. Likewise, Brunner et al. [48] found that polymorphic residues in the ARC2 domain that disrupt Pm3 function appear to be concentrated on one side of the ARC2 domain, and are largely solvent exposed, suggesting that they may be involved in intra- or intermolecular interactions. In the course of this chimeric protein analysis we generated some recombinant proteins that exhibited novel and expanded recognition specificities, through bringing together unique combinations of polymorphic sites. One of these (L6493L51193L6) was shown to function to induce an HR in tobacco, recapitulating the expanded recognition observed in Y2H assays. This correspondence between Avr: R interaction in yeast and HR induction in planta, now observed across a large number of L-AvrL567 pairwise combinations (Figure 1 and 7; [16], [45]) is consistent with the hypothesis of ligand interaction triggering signalling through competition with inhibitory intramolecular interactions. This suggests that recombination of existing polymorphisms through sequence exchanges is a powerful method for both generating changes in recognition specificity and fine-tuning the strength of defense response during evolution of resistance genes. This mechanism, along with induced mutation, may be adapted to engineering novel resistance genes that can be deployed in agriculture [53]. This process may require not only changes to LRR domain to generate new binding specificities, but also concomitant changes in N-terminal domains to optimise the defense signalling output. Site-directed mutants of AvrL567 were constructed using the Gene-Tailor kit (Stratagene) according to the manufacturer' s instructions. Chimeric L5–L6 proteins were constructed using native and introduced restriction sites, and/or by PCR-based fusion of overlapping sequences as described in Text S1 and Figure S5. Chimeric proteins were either constructed directly in pGADT7, or were sub-cloned in pBSK prior to construction in pGADT7. All constructs were checked by restriction enzyme digests and DNA sequencing. GAL4-binding domain (BD) fusions, and transcriptional activation domain (AD) fusions to L5, L6, L5–L6 chimeric proteins and to AvrL567 mutants and variants, were prepared in the pGBT9 and PGADT7 vectors (Clontech), as described [16], [45]. Yeast transformation, lacZ and His growth assays were performed as described in the Yeast Protocols Handbook (Clontech). Yeast proteins were extracted by the trichloro-acetic acid method, separated by SDS/PAGE, and transferred to nitrocellulose membranes (Pall) by electroblotting. Membranes were blocked with 5% skim milk and probed with anti-HA mouse monoclonal antibodies (Roche), followed by goat anti-mouse antibodies conjugated with horseradish peroxidase (Pierce). Labeling was detected with the SuperSignal West Pico chemiluminescence kit (Pierce). DNA constructs encoding AvrL567 proteins lacking the signal peptide, or full-length L5, L6 or L6493L51193L6 cDNAs, were cloned into the binary vector pTNotTReg between the cauliflower mosaic virus 35S promoter and ocs terminator sequences. Agrobacterium tumefaciens (GV3101-pMP90) cells containing these constructs were grown for 36 h at 28°C in LB media supplemented with appropriate antibiotic selections. Cells were pelleted, resuspended in infiltration medium (10 mM MgCl2,200 µM acetosyringone), adjusted to OD600 nm = 1 and incubated for 2 h at room temperature. Resuspended cells were infiltrated with a 1-mL needleless syringe into the leaves of near isogenic lines of flax plants containing L5, L6 (cv. Bison) or L6L11RV (cv. Ward), or into leaves of 3-week-old tobacco plants (W38). Transgenic tobacco expressing AvrL567-A was described by Dodds et al. [42]. The non-synonymous/synonymous rate ratio parameter ω was estimated using the program CODEML [54] in phylogenetic analysis by ML v. 4. 2. Tests for positive selection were performed using the site class models that estimate ω for amino acid sites. Neutral sites have ω = 1, those under purifying selection have ω<1, and those under positive selection have ω>1 [55]. Likelihood ratio tests were performed using lnL values from the models M7 and M8 by comparing the test statistic 2ΔlnL = 2 (lnLM7−lnLM8), with the χ2 distribution (d. f. = 2). For model M8, the empirical Bayes [55] procedure estimated the mean ω-value for each codon site, and the posterior probability that the site is under positive selection.

Title: Intramolecular Interaction Influences Binding of the Flax L5 and L6 Resistance Proteins to their AvrL567 Ligands Summary: The biotrophic fungus Melampsora lini is the causal agent of flax rust disease. Flax produces immune-receptor proteins that recognize fungal effector proteins, and subsequently signal the activation of plant defense responses. Here we report the molecular details of interactions between L-locus immune-receptors and AvrL567-locus effectors, as well as the engineering of an enhanced flax immune-receptor. In order to investigate the role of AvrL567 amino acid residues hypothesized to mediate interactions with L-locus immune receptors, we generated a series of site-direct mutations in AvrL567 proteins. Conversely, to investigate the role of regions hypothesized to mediate interactions with AvrL567 effectors, we generated a series of chimeric L-locus immune-receptors that contain swaps between, and within protein domains. Interactions between modified immune-receptors and effector proteins were evaluated using the yeast-two-hybrid system and transient expression in planta. Our results revealed that interactions between L-locus immune receptors and AvrL567-locus effector proteins involve multiple surfaces, and that intramolecular interactions between, and within, domains of L-locus immune-receptors plays a crucial role in these interactions. Finally, the generation of an enhanced immune-receptor is an important proof-of-concept demonstrating the utility of protein engineering in generating novel disease resistance in agricultural crops.

lay_plos

en

Summarize: Hollywood has Americanised Roald Dahl's characters and lost their charm Fantastic Mr Fox (PG) Fantastic Mr Fox is one of Roald Dahl's finest stories, about a wily fox outwitting three farmers determined to exterminate him and his family. The charm of Dahl's book is that it takes a dark, realistic view of foxes as predators, but the humans are even worse. The farmers, Boggis, Bunce and Bean, are as greedy and implacable as a bunch of bonus-crazed bankers, which makes it easy for children and adults alike to root for the family-oriented Mr Fox. Director Wes Anderson ( Rushmore, The Royal Tenenbaums, The Darjeeling Limited) goes with the kind of stop-motion animation pioneered by Wladyslaw Starewicz in The Tale Of The Fox (1930). But this has none of the charm of Wallace And Gromit, or even those irritating compare-the-meerkat commercials. Cunning plan: Mr Fox (George Clooney) and his family attempt to outsmart farmers Boggis, Bunce and Bean in Roald Dahl's classic story For children, the jerky movement of the animals may be disturbing, not to mention the moments when they bare their unbeautiful teeth. They look horribly like stuffed, dead animal heads on human bodies, with virtually no expression on their rigid little faces. Anderson, who lived in Paris while his film was shot in England and tried to direct the thing by email, has set the story in an annoyingly spaced-out vision of England, seemingly based on viewings of Postman Pat while under the influence of recreational drugs. Only the bad guys sound English. The good ones speak with American accents and embrace a U.S. lifestyle, hanging out at the Five and Dime and playing a baseball-type game with very American coaching methods. Dahl's happy family of foxes is turned by Anderson and his co-writer Noah Baumbach (The Squid And The Whale, Margot At The Wedding) into one of their typically hip but dysfunctional American families, with Mr Fox (voiced with maximum smugness by George Clooney) a newspaper columnist undergoing a midlife crisis in which he keeps reverting to his wilder youth. Jason Schwartzman, a charmless actor incomprehensibly over-indulged by Anderson, plays Fox's son as a needy, self-absorbed whinger. One of several new and thoroughly unessential characters is Mr Fox's nephew Kristofferson (Eric Anderson, the director's brother), a kick-boxing yoga enthusiast. The imposition of Anderson's perennial-adolescent obsessions on to Dahl's simple tale is itself dysfunctional, for it makes the story much less accessible to children, and indeed unsympathetic to anyone who loathes American psychobabble. Anderson's equally ill-advised attempt to turn the piece into a wisecracking Ocean's 11-style heist movie also strikes a dissonant chord, especially as the director's acting friends (such as Owen Wilson and Bill Murray) are bizarrely unsuited to a British context. The script isn't nearly as witty as Anderson thinks it is. Time and time again, would-be comic lines are tossed off as if they are Wildean epigrams (such as when Mr Fox tells his wife: 'You're still as fine-looking as a crème brulee'). But most sink like a Sharon Stone movie because they're not funny or perceptive. I am sure that Dahl himself would roll his eyes at the dreadfully immature attempts at adult sophistication, which come across as complacent, self-congratulatory whimsy. Too many of Anderson's numerous changes are pointless. The marital tension between Mr and Mrs Fox (a wasted Meryl Streep) doesn't come to anything, and Mr Fox's career as a newspaper columnist is an irrelevance. Whereas Dahl kept his foxes and other animals relatively true to their wild selves, Anderson humanises and Americanises without bothering to establish a coherent set of rules for his far too self-consciously quirky universe. Three splendid films have emerged from Roald Dahl's children's books: The Witches, Matilda and Charlie And The Chocolate Factory. And some people are fonder than I am of Willy Wonka And The Chocolate Factory, Danny The Champion Of The World and James And The Giant Peach. Fantastic Mr Fox has been weirdly overpraised, possibly because it opened the London Film Festival in a blaze of publicity for its celebrity cast, but it is by some distance the worst of the lot. Verdict: Fantastically disappointing Early in "Fantastic Mr. Fox," the youthful, wily Mr. Fox learns that Mrs. Fox (Meryl Streep) is pregnant. He vows to quit his life of crime and start making an honest living, the better to care for his new family. Fast-forward several fox years: Mr. Fox is now a newspaper columnist, and the dapper-yet-casual cords he used to wear appear to have been put in storage. Now his skinny arms, with their furry elbows, dangle from the awkwardly utilitarian short-sleeve shirts he now wears. Mrs. Fox is happy enough in the hole in the ground where the family now lives. But Mr. Fox wants more: He likes trees, grass, action. And so he leaves for work one day -- first casting a baffled glance at the superhero-cape and pants-tucked-into-socks outfit his weirdo son, Ash (Jason Schwartzman), is wearing -- and goes straight to see his lawyer, Badger (Bill Murray), to investigate the possibility of purchasing a new home inside a shady tree. There should be something incongruous about the sound of George Clooney's cashmere-flannel voice coming from the mouth of a somewhat rangy-looking fox in a country gent's corduroy suit: Why should a matinee idol suffer the indignity of being trapped in a puppet's body? But from the first minute of the Wes Anderson stop-motion-animated feature "Fantastic Mr. Fox," Clooney is that creature, the genuinely fantastic Mr. Fox of the title, a rapscallion charmer who wears many hats: husband, father, newspaperman, chicken thief. It's one thing for an actor to feel comfortable in his own skin; it's another for him to feel completely at home in the body of a fake-fur and metal-armature vulpus vulpus. And yet Clooney's naturalism is of a piece with the joyous, marvelously detailed movie around him, adapted from Roald Dahl's novel with adventurousness and seemingly boundless love by Anderson and Noah Baumbach. "Fantastic Mr. Fox" is possibly the finest picture about family, community and poultry thievery ever made. Clearly, it's midlife-crisis time for Mr. Fox, who not only moves his family into that new home but reenters a life of crime, enlisting the help of a spacey possum, Kylie (Wally Wolodarsky), to infiltrate the fortresses of three very rich, and very mean, local farmers: Boggis (Robin Hurlstone), Bunce (Hugo Guinness) and, the nastiest of them all, Bean (Michael Gambon). Before long Mr. Fox's mild-mannered, yoga-enthusiast nephew, Kristofferson (Eric Anderson), a gifted natural athlete, is also reluctantly pulled into the act, to the annoyance of the scrawny, disaffected kit Ash, who yearns to be a star at something and feels he's good at nothing. Dahl's novel is slim and exciting: It covers, in a very small number of pages, the Fox family's efforts to dig themselves deeper and deeper into the ground in order to escape the farmers, who have become obsessed with destroying them. Other woodland animals have also become displaced by the farmers' aggressive mania, and so Mr. Fox and his family, in addition to saving themselves, come to the aid of their homeless neighbors. They do this, of course, by stealing -- this is, after all, a Roald Dahl story, with all the vaguely disreputable pleasures that implies. There's a crotchety generosity to Dahl's work, too, which Anderson and Baumbach have captured perfectly here. To fill out a feature-length movie, they've had to expand upon and embroider Dahl's story, but they've done so without bloating the picture or overloading it. "Fantastic Mr. Fox" feels colloquial and modern: When we first meet Mrs. Fox, she's wearing an Indian-style tunic decorated with braid embroidery, a favorite hip-mom uniform among young mothers everywhere. And Mr. Fox, early in the film, explains apologetically, "I used to steal birds, but now I'm a newspaperman," perhaps a reflection not just on his lost, wild youth, but on the fact that he's moved on to a profession that's pretty much facing extinction itself. But "Fantastic Mr. Fox" also shows a sense of protectiveness toward the past, largely because of the somewhat rough-looking, slightly jerky (by digital-animation standards, at least), folk-art-style stop-motion animation technique Anderson has chosen. The puppets, with their kind-of-crazy eyes and even crazier whorls of fur, capture the spirit, if not necessarily the specifics, of the cheerful, mildly insane Quentin Blake illustrations that accompany Dahl's text. Anderson and his team (including, of course, a large crew of animators, as well as production designer Nelson Lowry and director of photography Tristan Oliver) pay a great deal of attention to texture and movement: The animals' fur swirls every which way, in accordance with their movements; even Mr. Fox's suits were reportedly modeled on the corduroy and tweed ones Anderson himself favors. (The puppet makers went so far as to obtain fabric swatches from Anderson's tailor.) Anderson also has a great deal of fun contrasting his fantasy foxes with their real-life counterparts in nature: His foxes walk and speak with the most elegant manners -- it doesn't hurt that the structure of their legs makes it look as if they're walking on tiny high-heels. They hover over exquisitely prepared platters of food, savoring the delicate blend of aromas -- and then descend upon them, suddenly realistically foxlike, snuffling and snarfling and sending food flying all over the place. There's so much to look at, and to giggle over, in "Fantastic Mr. Fox": It has style and wit and heart, without ever being overly whimsical, a trap Anderson has too often fallen into. "Fantastic Mr. Fox" could turn out to be the one movie Wes Anderson naysayers end up loving, and the one his loyal fans treat as a lesser accomplishment, a trifle. Anderson has always frustrated me: On the plus side, I can sense that he tries to work from the heart, and he certainly cares about craftsmanship. But nearly all of his movies, until now, have suffered from self-conscious quaintness, and their flat, homespun quirks have left me cold. "Fantastic Mr. Fox" is different: The story is a great canvas for Anderson's visual inventiveness (offering, for one thing, lots of opportunities for those cozy, dollhouse-style cross-section views he loves, showing various creatures going about their routine daily activities). It also revisits, in subtle and wonderful ways, many of Anderson's key themes, among them the prickly pleasures of being part of a jovial, like-minded community -- or of a mismatched family. In one of the movie's loveliest moments, the cousins Ash and Kristofferson, unable to breach their differences, fume and argue in their cramped, shared bedroom. Unable to sleep, they creep out of their beds to watch a tabletop model train clickety-clack around its track in the darkness, momentarily distracted and enthralled by the blinking colored lights and the soothing, tinny whir of its engine. I'm not sure I can explain why Anderson's trademark dry, clever patter seems less tortured, and so much funnier and more believable, when it's emerging from the mouths of animal puppets with scruffy, disarranged fur. But "Fantastic Mr. Fox" is one of the few recent movies I can think of that truly captures the vibe of a childhood spent largely with books. I'm not talking about the overrated notion of "being returned to a sense of childlike wonder," or anything like that. I'm talking about a movie that captures something even more intangible than that, the very texture of an experience: Looking at all the details in "Fantastic Mr. Fox" -- the character's wayward whiskers, their little vests, the mansionette hideaways they've dug for themselves in the ground -- brought back the quiet, intense joy I felt as a kid, first poring over illustrated details in picture books (the nooks and crannies of Beatrix Potter's rabbit warrens and mouse houses, for example) and later in the semi-fanciful, semi-naturalistic details to be found in Kenneth Grahame and A.A. Milne and Dahl. "Fantastic Mr. Fox" is an intricately detailed and accomplished piece of work. (It amazes me that 2009 has brought us not just one but two dazzling stop-motion-animated pictures, the other being Henry Selick's gorgeous and spooky "Coraline," adapted from Neil Gaiman's equally terrific children's novel.) And yet what's wonderful about it is how casual and free, how un-fussed-over, it feels. Anderson is clearly taking a stand against the strained realism (make that "so-called realism") of digital animation, up to and including the repellent motion-capture technology that has turned the once-fine filmmaker Robert Zemeckis into a zombie. And in the end, Anderson's picture is more wondrous in the ways that count, more palpably believable within its fantasy world, than anything Dreamworks and -- yes, I'll say it -- Pixar (with the notable exception of Brad Bird's projects) has come up with. As a work of animation, and of art, "Fantastic Mr. Fox" is wily, clever and mischievous, without ever being too arch or knowing. It also has the distinct aura of something that's been made entirely by hand with care and affection -- a few misshapen nubs here and there only add to the charm. Anderson has pulled off the most elusive of goals: He's made a nonchalant masterpiece, a movie that feels dog-eared and loved before it's even reached our hands. Film Reviews Fantastic Mr. Fox -- Film Review "Fantastic Mr. Fox" Rate This Film 61 2 User Reviews » See all » Write a Review » See all » Leave a Comment Cast and Crew Executive Producer: Steven Rales Executive Producer: Arnon Milchan Producer: Allison Abbate Producer: Scott Rudin Producer: Wes Anderson Producer: Jeremy Dawson Director: Wes Anderson Screen Writer: Wes Anderson Screen Writer: Noah Baumbach Animation Director: Mark Gustafson Director of Photography: Tristan Oliver Editor: Ralph Foster Editor: Stephen Perkins Prod. Designer: Nelson Lowry Music: Alexandre Desplat Cast: George Clooney ( Mr. Fox ), Meryl Streep ( Mrs. Fox ), Jason Schwartzman ( Ash ), Bill Murray ( Badger ), Wally Wolodarsky ( Kylie ), Michael Gambon ( Franklin Bean ), Willem Dafoe ( Rat ), Owen Wilson ( Coach Skip ), Jarvis Cocker ( Petey ) Bottom Line: Although sometimes too sly for its own good, this great-looking carnivorous caper brings Wes Anderson's whimsical melancholy to a kids' classic. The title character of Wes Anderson's first animated feature is a cad, a rogue, an incorrigible thief. He's also a family man, struggling to do right on the marital and child-rearing fronts. Voiced by George Clooney, he's no small part Danny Ocean. If "Fantastic Mr. Fox" is an eye-popping, kid-friendly adaptation of Roald Dahl's attitude-packed 1970 book, it's at least as much a film for adults.The screenplay sometimes overdoes the winking asides, and the film doesn't so much flow as jump from one set piece to the next. But with animation director Mark Gustafson, DP Tristan Oliver and production designer Nelson Lowry, Anderson has created a world as stylized and inventive as anything he's done. From the fox-red glow of a morning idyll to the noirish gutter scene where one character meets his end to the icy fluorescent glare of the film's closing scene -- happy but not without compromise -- "Fox" is a visual delight.The movie, which premiered at the London fest and bows Nov. 13 in New York and Los Angeles before going wide for Thanksgiving, is not likely to dethrone Disney's "Princess" in the year-end animation tallies, but word-of-mouth should make it a cunning alt-family-fare contender.The word "texture" gets tossed around a lot, but this stop-motion escapade is alive with it, beginning with the puppets by U.K. outfit MacKinnon and Saunders. They're furry animals who walk upright and dress with style but who are, we're reminded on more than one occasion, wild creatures. They kill to survive, and when they eat, they devour. Miss Manners would not approve.Among Anderson's films, "Fox" hews closest to "The Life Aquatic With Steve Zissou" in its handmade aesthetic, though its vision is far less, well, fantastic. That strange adventure, like this film, was co-written by Anderson and Noah Baumbach. Their "Fox" script softens the dysfunctional edges without sugarcoating the director's ideas about the nuclear family and conflicted father figures. There's plenty of angst, of the grown-up and teen varieties, to go around. Just listen to the way Mrs. Fox (Meryl Streep), the antihero's former partner in crime, says "I'm pregnant."Twelve "fox years" after she makes that ambivalent statement, Fox has forsaken larceny and works as a newspaperman who suspects that no one reads his column. (Who says kids' films can't reflect contemporary reality?) Wanting to move on up as a homeowner, he buys a tree he can't quite afford. Soon he's donning the raffish corduroy getup from his bad old days and enlisting the help of a spacey opossum (Wally Wolodarsky) in "one last" big job. His targets are the three nastiest farmers for miles around: Boggis, Bunce and the ultramean Bean (Michael Gambon).As deliciously dry as Gambon's villain is, the bad guys are of less concern to Anderson than they were to Dahl. What drives this caper is the Fox home front. Diminutive son Ash (Jason Schwartzman, in a fine adolescent sulk) can't measure up to his debonair dad or his perfect cousin Kristofferson (an equally effective Eric Anderson, brother of the director). A terrific detail is the way Ash twitches his ear when his feelings are hurt. The cousins' rivalry is comic and touching, and Anderson administers a familiar Hollywood lesson -- the beauty of being different -- without the usual schmaltz.Star voice casts can be more distracting than helpful, but Clooney and Streep bring shadings to their characters that deepen the story. The supporting cast, effective if not indelible, includes Anderson regulars Bill Murray as attorney Badger, Willem Dafoe as security guard Rat and Owen Wilson as Ash's discouragement-dispensing coach.Boomer faves by the Beach Boys and the Stones punctuate the soundtrack, with Alexandre Desplat delivering an elegant gallop of a score. Upping the hipster quotient in lovely, non-ironic fashion is Jarvis Cocker's bit as farmhand Petey, a banjo-playing Jarvis Cocker look-alike who strums a rootsy ditty by campfire.

Summary: Wes Anderson is winning raves for his stop-motion take on Roald Dahl's classic Fantastic Mr. Fox, with voices by a raft of stars including Meryl Streep and George Clooney: It's so good, David Edelstein forgot himself. When the flick ended, he writes in New York, he wished "the whole battery of gifted artists could come out and take a bow." The film "feels handmade, present, as if they're all backstage and the curtain is going up before your eyes." Bravo! writes Stephanie Zacharek for Salon. It tops "anything Dreamworks and-yes, I'll say it-Pixar (with the notable exception of Brad Bird's projects) has come up with." Sheri Linden was skeptical at first. "Star voice casts can be more distracting than helpful," she writes in the Hollywood Reporter, but she came around. "Clooney and Streep bring shadings to their characters that deepen the story." A rare eh comes from Chris Tookey of the Daily Mail, who says the flick "has been weirdly overpraised." Kids may be disturbed by the "jerky movement" of what look like "stuffed, dead animal heads on human bodies."

multi_news

en

Summarize: CROSS REFERENCE TO RELATED APPLICATIONS This application is a continuation-in-part of application Ser. No. 09/862,939 filed May 22, 2001, for a “Spinner Bait Tackle Organizer with Compression Catches,” now U.S. Pat. No. 6,860,059. BACKGROUND OF THE INVENTION Fishing enthusiasts consistently face the problem of baits and lures becoming tangled in tackle boxes. Fishing hooks, lures, and various kinds of baits are often made with thin strips of plastic or metal that tend to intertwine with one another, hindering quick retrieval of one piece of equipment from the box. To alleviate this problem, some fishermen divide their tackle boxes into compartments that allow the fishermen to gather similar lures and hooks into a designated section of the container. Within each section of the container, however, lures and hooks still twist around each other and must be manually separated before use. Previous tackle boxes and containers have been developed in efforts to alleviate the problem of tangled lures and hooks. U.S. Pat. No. 2,220,817 (Holmes, 1939) illustrates the use of grooved slots in a tackle container to separate individual hooks and lures. The Holmes tackle container incorporates a plurality of panels attached to the bottom of the container along a central line so that the panels open outwardly like a fan. Each panel has a series of slots extending vertically from the top edge of the panel into the body of the panel. Either a hook or any curved portion of a fishing lure may slide within one of the slots in the panel to suspend the lure from that slot. The panels therefore provide a multitude of slots to separate individual fishing lures. Separating fishing lures via grooved slots in tackle container panels is also shown in U.S. Pat. No. 5,606,820 (Suddeth, 1997). The Suddeth patent is designed to organize and hold fishing lures having treble hooks hanging from the underside of the lure. Suddeth provides multiple panels within the container that may be extended vertically from a horizontal position. When the panels are vertically extended, the hooks from the underside of a fishing lure slide into grooves in the panels. The panels are then returned to their horizontal position so that the body of the fishing lure rests on the groove with the hooks hanging underneath the panel. The treble hooks of each lure are separated from the hooks of other lures to prevent tangling. Suddeth also shows a panel in a tackle container having retainer bars from which the treble hooks of the lure may hang to suspend the lure alongside the panel. U.S. Pat. No. 4,958,730 (Bunten, 1990) shows a tackle box with a plurality of deep storage drawers that slide within an enclosure. The drawers include attachment points from which lures and hooks may be suspended. Depending on the kind of lure, the lure may be positioned so that it is suspended from the top of one of the drawers in a vertical direction. Alternatively, the lure may lie flat in the bottom of a horizontal drawer. For lures that should hang vertically, the storage drawer includes an elastic retaining member supported within the top section of the drawer. U.S. Pat. No. 5,228,232 (Miles, 1993) also shows a tackle container with divided compartments in which lures and hooks may be organized. Each compartment within the Miles &#39;232 patent incorporates a groove within one of the sidewalls of each compartment. A portion of a fishing lure, such as a wire neck or a thin hook, slides within the groove to hold the fishing lure in place within its compartment. The groove within the compartment sidewall may extend from the top of the sidewall to a midpoint toward the bottom of the container. Other inventions in the art of tackle containers emphasize individual hooks and ways of preventing the hooks from becoming tangled. U.S. Pat. No. 3,133,374 (Benson, 1964) uses various upright hook holders to individually retain fishing hooks within a container. The Benson &#39;374 patent provides a pair of vertical wire stems that allow a hook to be positioned between the stems. In operation, the Benson hook organizer supports the hook by looping the eye of a hook around one stem and attaching the curved, pointed end of the hook to an opposite stem. The stems of the Benson hook organizer are positioned with enough tension between the two to hold the hook horizontally across the base of the Benson tackle container. U.S. Pat. No. 6,101,760 (Garman, 2000) provides another mechanism for holding hooks and preventing tangling. The Garman hook organizer also allows easy retrieval of one hook without touching other hooks in the container. The Garman &#39;760 patent shows an organizer in which fishhooks of various sizes may be removably attached between two parallel edges. The edges of the modular hook organizer include a sequence of notches into which the fishhook is pushed in a forced fit. In a typical embodiment of the Garman &#39;760 patent, the shank of a fishhook is held within a pair of notches in two substantially parallel vertical walls of the hook organizer. When installed within the notches, the fishhook is elevated from the bottom of a horizontal portion of the hook organizer. The curved hook end of the fishhook is also snapped into a notch in the horizontal portion of the hook organizer. The Garman &#39;760 patent, therefore, discloses a hook organizer that provides at least three contact points for a fishhook-two notches within vertical walls of the organizer and one notch within a horizontal floor. Each organizer in the Garman &#39;760 patent may have connecting elements that allow the user to connect more than one organizer in a modular fashion. Another example of tackle containers providing individualized holders for specialized baits and lures is U.S. Pat. No. 6,256,925 (Blackburn &#39;925). The Blackburn &#39;925 patent uses individual enclosures to hold each lure and is particularly useful for holding spinner baits. Each individual enclosure is an upright rectangular box with an extra cavity formed in the top of the box by a triangular attachment extending from the front wall of the enclosure. The triangular attachment includes a groove pointing downward toward the bottom of the box. A spinner bait fits within the enclosure with one segment of the spinner bait extending downward into the upright box as a second segment of the spinner bait fits across the triangular attachment at the top of the enclosure. The second segment of the spinner bait is compressed toward the segment in the box so that the second segment fits within the groove in the triangular attachment. The spinner bait is held within the enclosure by a force of compression between the two segments. U.S. Pat. No. 4,245,422 (Souza, 1981) discloses a tackle box with a pullout stacking tray structure in which the individual trays are connected to the walls of the container by pivoting links. The container includes vertically hanging attachment walls on the sides of the stack of trays. Each attachment wall includes a plurality of projecting screws from which fishing lures may hang. In one embodiment of the Souza &#39;422 patent, three projecting screws are arranged in a triangle. A spinner bait lure may be balanced at the elbow between lure segments so that one segment is supported by a lower screw as the opposite segment rests on a screw directly across from the lower screw. U.S. Pat. No. 5,960,582 (Wilkins, 1999) shows another vertically hanging attachment wall within a tackle container for securing spinner bait lures. The Wilkins &#39;582 patent uses a screw that projects from one wall of the container to balance a spinner bait at the elbow between lure segments. The weight of a spinner bait lure is unequal on opposite sides due to different components attached to each segment. The weight difference on either side can possibly lead to the lure sliding around and falling off the projecting screw. The Wilkins &#39;582 patent incorporates retaining walls on either side of the projecting screw to stop movement of the spinner bait lure and retain the lure in a convenient position. SUMMARY OF THE INVENTION It is an object of the present invention to provide a tackle container that is capable of storing different kinds of fishing lures and baits within a single container. It is another object of the present invention to provide a tackle container with a structure that secures lures and baits with segmented bodies inside the container so that the container may be carried and moved without tangling the lures and baits. It is yet another object of the present invention to provide removable inserts that fit within the tackle container of this invention to provide a storage structure for different kinds of fishing lures and baits. The invention herein meets these objects by providing a tackle container that incorporates appropriate inserts therein to hold segmented lures in a fixed position. The inserts comprise a lure retainer with notched posts that secure one segment of the lure while a second segment of the lure lies on the base of that insert. The inserts are particularly structured to accommodate spinner bait lures and buzz bait lures within a tackle container to prevent tangling of the lures therein. The inserts are interchangeable so that one container may be used to store a single kind of lure or different kinds of lures at the option of the user. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a plan view of a tackle container according to the invention herein with inserts installed on either side of a partition. FIG. 2 is a plan view of a spinner bait insert according to the invention herein. FIG. 3 is a top view of the spinner bait insert according to the invention herein. FIG. 4 is a front side view of the spinner bait insert according to the invention herein. FIG. 5 is a sectional view of the lure retainer post of a spinner bait insert according to the invention herein with a spinner bait lure installed in the uppermost notch. FIG. 6 is a sectional view of the lure retainer post of a spinner bait insert according to the invention herein with a spinner bait lure installed in the lower notch of the post. FIG. 7 is a plan view of the buzz bait insert according to the invention herein. FIG. 8 is a top view of the buzz bait insert according to the invention herein. FIG. 9 is a front side view of the buzz bait insert according to the invention herein. FIG. 10 a is a plan view of a buzz bait lure to be installed in the lure retainer post of a buzz bait insert according to the invention herein. FIG. 10 b is a plan view of a buzz bait insert with a buzz bait lure installed within the lure retainer post and the corresponding post of the insert. FIG. 11 is a plan view of an open tackle container that is structured to receive the inserts of the invention herein. FIG. 12 is a plan view of a tackle container according to the invention herein with a spinner bait insert and a buzz bait insert interchangeably installed therein. FIG. 13 is a plan view of a buzz bait insert according to the invention herein wherein the buzz bait lure is held within buzz clips attached to the base of the insert. DETAILED DESCRIPTION FIG. 1 shows inserts ( 5, 10 ) for placing inside a fishing lure container ( 25 ) to hold lures having segmented bodies. Fishing lures with segmented bodies include, but are not limited to, spinner baits ( 6 ) and buzz baits ( 9 ). A typical spinner bait, as shown in FIG. 2, comprises a V-shaped wire portion with an elbow ( 7 ) at the vertex that allows for compression of the wires without causing deformation of the lure. The elbow ( 7 ) is also known as an R-bend. A buzz bait, as shown in FIG. 7, is similarly made of a wire that is bent at an elbow ( 11 ) separating two segments. A typical fishing lure also has a head ( 12, 18 ), a hook ( 13, 19 ), a neck ( 14, 20 ), a skirt ( 16, 22 ) and a blade ( 15, 21 ). Lures may be organized and secured in a container ( 25 ) that is conveniently carried or otherwise transported during use. Apparatuses for organizing and securing segmented lures in convenient containers are shown in FIGS. 2 and 7. The apparatuses are inserts for placing inside fishing lure containers ( 25 ) to hold lures having segmented bodies ( 6, 9 ). According to the invention herein, inserts for holding segmented fishing lures each include a base ( 31, 46 ) and a lure retainer ( 32, 47 ) connected to the base ( 31, 46 ). The base ( 31, 46 ) may be substantially flat (i.e., planar). The lure retainers ( 32, 47 ) each have an upright, substantially perpendicular post ( 34, 50 ) with notches ( 35, 51 ) defined within the post. The lure retainers ( 32, 47 ) further include channel barriers ( 38, 54 ) extending substantially perpendicularly from each post ( 34, 50 ) across a respective base ( 31, 46 ) of an insert ( 30, 45 ). The post ( 34, 50 ), the channel barrier ( 38, 54 ), or both may be connected to the base ( 31 ) of the insert ( 5, 10 ). Each notch ( 35, 51 ) in a respective post ( 34, 50 ) secures one segment of a lure ( 6, 9 ) above the base ( 31, 46 ) of the insert ( 30, 45 ). The lure retainer ( 32, 47 ) positions the lure so that a second segment of the lure ( 6, 9 ) rests on the base ( 31, 46 ) of the insert ( 30, 45 ) alongside the channel barrier ( 38, 54 ). The unique arrangement of a base ( 31, 46 ), a lure retainer ( 32, 47 ), a notched post ( 34, 50 ), and a channel barrier ( 38, 54 ) provides an insert that is highly suitable for securing fishing lures with segmented bodies inside a portable container ( 25 ). One insert ( 30 ) according to the invention herein is particularly useful for holding spinner baits ( 6 ) in a container ( 25 ). FIGS. 2-6 illustrate the spinner bait insert. The spinner bait insert ( 30 ) has a post ( 34 ) with the notch ( 35 ) defined within the side edge of the post ( 34 ). The notch ( 35 ) preferably has an upward orientation from the side of the post ( 34 ) toward the top edge of the post ( 34 ). The post may define more than one notch ( 36 ) in the side edge as necessary to accommodate lures of varying sizes. The spinner bait insert ( 30 ) includes the notch ( 35 ) in the side edge of the post ( 34 ) so that a spinner bait lure ( 6 ) may be compressed between the notch ( 35 ) and the base ( 31 ) of the insert ( 30 ). As shown in FIGS. 5 and 6, when a spinner bait lure ( 6 ) is positioned within the lure retainer ( 32 ) by compressing an upper segment of the lure ( 6 ) into the notch ( 35 ) in the side of the post ( 34 ), a lower segment of the spinner bait lure lies along the base ( 31 ) beside the channel barrier ( 38 ). The channel barrier ( 38 ) of the spinner bait insert ( 30 ) extends substantially perpendicularly from the lure retainer post ( 34 ). In one embodiment the channel barrier ( 38 ) extends outwardly from near the top of the post ( 34 ) and connects to the base ( 31 ) of the insert ( 30 ). The channel barrier ( 38 ) also may include a triangular shaped gusset ( 41 ) that provides support between the post ( 34 ) and the base ( 31 ) of the insert ( 30 ). A spinner bait insert ( 30 ) may incorporate a plurality of lure retainers ( 32 ), positioned side-by-side, with each lure retainer ( 32 ) being connected to the base ( 31 ) of the insert ( 30 ). The side-by-side lure retainers ( 32 ) form a series of channels between adjacent channel barriers ( 38 ). In this configuration, one insert ( 30 ) may hold multiple spinner bait lures ( 6 ). Each spinner bait lure ( 6 ) is compressed so that one lure segment fits within a notch ( 35 ) in a respective lure retainer post ( 34 ), and another segment of that lure ( 6 ) lies within a channel formed by adjacent channel barriers ( 38 ). The notch ( 35 ) holds the lure in place by a downward compressive force, and the channel barriers ( 38 ) prevent the lure from moving side to side. The insert ( 30 ) provides a mechanism for organizing spinner bait lures ( 6 ), securing the lures in a fixed position, and maintaining the lures in an organized arrangement when the user transports the lures in a portable container ( 25 ). A spinner bait insert ( 30 ) with a plurality of side-by-side lure retainers connected to the base ( 31 ) may include neck rests ( 39 ) connecting adjacent lure retainers. The neck rests ( 39 ) provide support to the necks ( 14 ) of spinner bait lures ( 6 ) when the lure segments are compressed between the notch ( 35 ) of a lure retainer post ( 34 ) and the base ( 31 ) of the insert ( 30 ). The neck rests ( 39 ) may be formed into any convenient shape, but U-shaped neck rests ( 39 ) are useful for securing rounded lure necks ( 14 ). The inserts of this invention may also be configured to efficiently organize buzz bait lures ( 9 ) within a fishing tackle container. FIGS. 7-9 show the details of the buzz bait insert. A buzz bait insert ( 45 ) includes certain elements in common with the above-described spinner bait insert ( 30 ). In this regard, the buzz bait inserts each include a base ( 46 ) and a lure retainer ( 47 ) connected to the base ( 46 ). The lure retainers ( 47 ) also each have a substantially perpendicular post ( 50 ) defining a notch ( 51 ) within the post ( 50 ). The buzz bait lure retainers ( 47 ) further include channel barriers ( 54 ) extending substantially perpendicularly from each lure retainer post ( 50 ). In addition to the lure retainer post ( 50 ), the buzz bait insert ( 45 ) has a corresponding post ( 55 ) attached to the base ( 46 ) across from the lure retainer post ( 50 ). The corresponding post ( 55 ) may be connected substantially perpendicularly to the base ( 46 ) of the insert. The corresponding post ( 55 ) also defines a notch ( 56 ) within the corresponding post ( 55 ). In a preferred embodiment, the top edge of the lure retainer post ( 50 ) and the top edge of the corresponding post ( 55 ) define the respective notches in the post ( 50 ) and the corresponding post ( 55 ). The notches ( 51, 56 ) in the lure retainer post ( 50 ) and the corresponding post ( 55 ) are of a size and shape that allow a segment of a buzz bait lure ( 9 ) to fit therein. In one preferred embodiment, the notches ( 51, 56 ) are generally shaped like a keyhole extending from the top edge into the bodies of the post ( 50 ) and the corresponding post ( 54 ). The notches in the post and corresponding post ( 51, 56 ) may be coaxial to allow the segment of the buzz bait lure to fit therein in a linear position. One segment of the buzz bait lure ( 9 ) fits within the respective notches ( 51, 56 ) of the post ( 50 ) and the corresponding post ( 55 ) such that the lure segment extends across the channel barrier ( 54 ). While one segment of the buzz bait lure ( 9 ) is held within the notches ( 51, 56 ) of the insert ( 45 ), the lure ( 9 ) is allowed to rotate so that a second segment of the lure ( 9 ) lies alongside the channel barrier ( 54 ) on the base ( 46 ) of the insert. In one preferred embodiment, the channel barrier ( 54 ) extends from the lure retainer post ( 50 ) and connects to the corresponding post ( 55 ). The channel barrier ( 54 ) may also connect to the base ( 46 ) as necessary for support. The buzz bait insert ( 45 ) may also include triangular-shaped gussets ( 57, 58 ) attached to the sides of the post ( 50 ) and the corresponding post ( 54 ) opposite the channel barrier ( 54 ). Each gusset ( 57, 58 ) adds support to a respective post ( 50 ) and corresponding post ( 55 ). A buzz bait lure insert according to the invention herein may also comprise a plurality of buzz bait lure retainers ( 47 ) positioned side-by-side with each lure retainer ( 47 ) being connected to the base ( 46 ). The side-by-side buzz bait lure retainers ( 47 ) form respective channels between each pair of adjacent channel barriers ( 54 ). Each buzz bait lure retainer ( 47 ) holds one segment of the buzz bait lure ( 9 ) within the notch ( 51 ) in the post ( 50 ) and allows a second segment of the buzz bait lure ( 9 ) to rotate and lie in a channel formed by adjacent channel barriers ( 54 ). A buzz bait lure insert having a plurality of buzz bait lure retainers ( 47 ) may also include a plurality of corresponding posts ( 55 ) positioned side-by-side and connected to the base ( 46 ). Each of the corresponding posts ( 55 ) is aligned across the base ( 46 ) from the post ( 50 ) of a respective lure retainer ( 47 ). The corresponding posts ( 55 ) each define a notch ( 56 ) within the corresponding post ( 55 ) so that the lure retainer post ( 50 ) and the corresponding post ( 55 ) hold one segment of a lure within their respective notches ( 51, 56 ) as another segment of the lure lies within the channel between adjacent channel barriers ( 54 ). As shown in FIG. 8, a buzz bait lure insert ( 45 ) according to the invention herein may include additional features that provide more support and add rigidity to the insert. In this regard, the insert ( 45 ) may have a support ridge ( 48 ) extending along the base ( 46 ) between each lure retainer ( 47 ). A second support ridge extends along the base ( 46 ) between each corresponding post ( 55 ). A similar third support ridge may extend horizontally on the inner surface of each end wall ( 60, 65 ). The tackle container inserts according to the invention herein may also include an end wall ( 60 ) connected to the base ( 31, 46 ) substantially parallel to the channel barrier ( 38, 54 ). The end wall ( 60 ) may include a substantially vertical rail ( 62 ) opposite the base ( 31, 46 ) on the outside of the end wall ( 60 ). The end wall ( 60 ) may also include a second substantially vertical rail ( 63 ) opposite the base ( 31, 46 ) on the outside of the end wall ( 60 ). The end wall ( 60 ) and its associated rails ( 62, 63 ) assist in securing the insert ( 30, 45 ) within a container ( 25 ). The insert may include a second end wall ( 65 ) connected to the end of the base ( 31, 46 ) opposite the first end wall ( 60 ). Similar to the first end wall ( 60 ), the second end wall ( 65 ) may have first and second vertical rails ( 67, 68 ) opposite the base of the insert on the outside the second end wall ( 65 ). Alternatively, the two end walls ( 60, 65 ) and the base ( 31, 46 ) of the insert may be formed as a single piece U-shaped construction. The inserts, described herein for organizing various kinds of fishing lures, conveniently fit within tackle containers for carrying multiple lures without tangling the lures. The invention, therefore, encompasses a container, as shown in FIG. 11, that utilizes the above-described inserts for holding and organizing fishing lures with segmented bodies. The container ( 75 ) has a bottom ( 78 ) with a continuous sidewall ( 79 ) connected to the bottom ( 78 ). The inserts, particularly the above-described spinner bait and buzz bait inserts ( 30, 45 ), removably fit within the continuous sidewall ( 79 ). The sidewall ( 79 ) of a container ( 70 ), designed to hold fishing lure inserts according to the invention herein, includes a plurality of pairs of substantially vertical ribs ( 82, 83 ) that define pairs of aligned slots ( 87, 88 ) across the bottom ( 78 ) of the container ( 75 ) from one another. As described above, the inserts have at least one end wall ( 60 ) connected to the base ( 31, 46 ) of the insert ( 30, 45 ). The end wall ( 60 ) has a substantially vertical rail ( 62 ) located on the outside of the end wall ( 60 ) opposite the base ( 31, 46 ) of a respective insert ( 30, 45 ). The vertical rail ( 62 ) slides between a pair of substantially vertical ribs ( 82, 83 ) on the sidewall ( 79 ) of the container ( 75 ), such that the rail ( 60 ) fits within the slot ( 87 ) formed by the substantially vertical ribs ( 82, 83 ). The inserts according to this invention may have multiple rails ( 62, 63, 67, 68 ) located on the outside of first and second end walls ( 60, 65 ). The container ( 70 ) is adapted to receive the inserts ( 30, 45 ) described herein and hold the inserts securely in place. The pairs of rails ( 62, 63, 67, 68 ) on each end wall ( 60, 65 ) removably slide between corresponding pairs of ribs ( 82, 83 ) on the sidewall ( 79 ) to hold the insert in place within the container ( 70 ). In one preferred embodiment, the container ( 75 ) may include a partition ( 90 ) extending between opposite sections of the sidewall ( 79 ). The partition ( 90 ) divides the container ( 75 ) into two portions. In this embodiment, at least one insert removably fits within each section of the divided container ( 75 ). The different sections of the container ( 75 ) are available to install different kinds of inserts in one container ( 75 ). For example, as shown in FIG. 12, one section of the container ( 75 ) may accommodate a buzz bait insert ( 45 ) while an adjacent section of the container ( 75 ) holds a spinner bait insert ( 30 ). A single container ( 75 ) may then be used to organize and carry multiple kinds of lures at once. In this regard, the inserts are interchangeable within the container. The container ( 75 ) may include one kind of insert on both sides of the partition ( 90 ), or it may include different inserts on either side of the partition. The embodiment of FIG. 12 shows the above-described container ( 75 ) fitted with an insert for holding fishing lures that have a neck, a lower lure segment, and an upper lure segment. The assembly includes a bottom ( 78 ), a continuous sidewall ( 79 ) connected to the bottom ( 78 ), and an insert ( 30 ) that removably fits within the sidewall ( 79 ). In one embodiment, the insert ( 30 ) is particularly suitable for holding and organizing spinner bait lures. The spinner bait lure insert ( 30 ) includes a base ( 31 ) and a lure retainer ( 32 ) connected to the base ( 31 ). The lure retainer ( 32 ) is a substantially perpendicular structure that includes a post ( 34 ) with a notch ( 35 ) formed in the post ( 34 ). The lure retainer ( 32 ) further includes a channel barrier ( 38 ) extending substantially perpendicularly from the post ( 34 ). The insert ( 30 ) may include all the features described above, which will not be repeated but that are incorporated as if set forth fully herein. The container ( 75 ) may include vertical ribs ( 82 - 85 ) along the sidewall ( 79 ) for receiving rails ( 67, 68 ) positioned on the outside of the end walls ( 60, 65 ) of the spinner bait lure insert ( 30 ). When assembled in this manner, the base ( 31 ) of the spinner bait insert ( 30 ) is flush against the bottom ( 78 ) of the container ( 75 ). The container lid ( 76 ) closes over the insert ( 30 ) to protect the contents therein. The lid ( 76 ) is attached to the container by hinges that are well known in the art of portable containers. The container lid ( 76 ) includes a fastening mechanism, including but not limited to, hinged handles that snap into place around the perimeter of the lid ( 76 ). FIG. 12 also shows an assembled fishing lure container according to this invention that includes the buzz bait insert ( 45 ) installed therein. Again, the container ( 75 ) includes a bottom ( 78 ), a continuous sidewall ( 79 ) connected to the bottom ( 78 ), and an insert ( 45 ) that removably fits within the sidewall ( 79 ). A buzz bait insert ( 45 ) installed in the container ( 75 ) has a base ( 46 ) and a lure retainer ( 47 ) connected to the base ( 46 ). The lure retainer of the buzz bait insert ( 45 ) is adapted to organize and hold buzz bait fishing lures within a notch ( 51 ) in a post ( 50 ) extending perpendicularly from the base ( 46 ). The buzz bait insert ( 45 ) also has a corresponding post ( 55 ) attached to the base ( 46 ) across from the post ( 50 ) of the lure retainer ( 47 ). A channel barrier ( 54 ) extends from the post of the lure retainer ( 47 ) across the base ( 46 ) and connects to the corresponding post ( 55 ). The corresponding post ( 55 ) defines a notch ( 56 ) therein for holding one segment of the buzz bait fishing lure. In operation, one segment of a buzz bait fishing lure fits within the respective notches of the post ( 50 ) and the corresponding post ( 55 ) such that the lure segment extends across the channel barrier ( 54 ) of the buzz bait insert ( 45 ). When one segment of a buzz bait fishing lure is positioned across the channel barrier ( 54 ) and resting within the notches of the lure retainer post ( 50 ) and the corresponding post ( 54 ), a second segment of the lure lies alongside the channel barrier ( 54 ). FIG. 13 shows yet another embodiment of an insert ( 95 ), according to the invention herein, that is particularly useful for holding fishing lures having segmented bodies. This embodiment fits within a container ( 75 ) as previously described. The container ( 75 ) has a continuous sidewall ( 79 ) connected to a bottom ( 78 ). The insert ( 95 ) that fits within the continuous sidewall ( 79 ) has a base ( 98 ) and a plurality of pairs of buzz clips ( 102, 103 ) attached to the base ( 98 ). A buzz clip ( 102, 103 ) has two contoured upright braces ( 105, 106 ) positioned immediately next to one another. The contoured braces ( 105, 106 ) are essentially mirror images and include rounded inner edges facing each other. The rounded inner edges are positioned to form a holding cavity ( 108 ) there between. The contoured braces ( 105, 106 ) clamp together with sufficient force to remain in a closed position until an opposite force pries the braces apart. A lure segment fits between the braces ( 105, 106 ) and will remain in the holding cavity ( 108 ) when the braces are in a closed position. A buzz bait insert ( 95 ) according to the invention herein has at least one pair of buzz clips ( 102, 103 ) attached to the base ( 98 ) of the insert ( 95 ). The buzz clips ( 102, 103 ) are aligned directly across from one another. A buzz bait lure segment may be secured within the holding cavity ( 108 ) of a first buzz clip ( 102 ) and extend across the base ( 98 ) of the insert ( 95 ) so that a second end of the buzz bait lure segment is secured within the holding cavity of an opposite buzz clip ( 103 ). In a preferred embodiment, the buzz clips in any given pair include coaxial holding cavities ( 108, 109 ) so that the pair of buzz clips ( 102, 103 ) open and close along a common axis. A buzz bait lure segment placed within a pair of buzz clips ( 102, 103 ) is free to rotate within the pair of holding cavities so that a second segment of the lure lies along the base ( 98 ) of the insert ( 95 ). In the drawings and specification, there have been disclosed typical embodiments on the invention and, although specific terms have been employed, they have been used in a generic and descriptive sense only and not for purposes of limitation, the scope of the invention being set forth in the following claims.

Summary: The invention is a tackle container that incorporates interchangeable inserts. The inserts secure different kinds of fishing lures and baits. For example, the inserts hold segmented lures within notched upright posts. In this regard, the inserts may be suitably structured to hold spinner bait lures, buzz bait lures, or other fishing lures with segmented bodies. Moreover, the inserts are interchangeable to facilitate a wider variety of uses for the tackle container.

big_patent

en

Summarize: TECHNICAL FIELD The present invention is generally related to restoration of eyelid function and correction of ptosis. BACKGROUND OF THE INVENTION A drooping of the upper eyelid below its normal level resulting in narrowing of the palpebral opening is known in ophthalmology as Ptosis. The muscle controlling lid opening is the levator palpebrae superioris and it is innervated by the third cranial nerve (oculomotor nerve). Ptosis may be congenital or acquired. In congenital ptosis the levator palpebrae superioris muscle that elevates the lid is either absent or not well developed. Acquired ptosis, on the other hand, is usually due to either diseases or injuries of the nerves that control the movements of the levator palpebrae superioris muscle. Ptosis may further be classified as myogenic, aponeurotic, neurogenic, mechanical or traumatic. The treatment of ptosis has traditionally required accurate and consistent evaluation and measurement as well as skillful use of surgical techniques to implement a functional and aesthetic correction. In most cases, surgery has been required to correct a ptotic eyelid. The surgical procedures used generally depend on the severity of ptosis and are described in the following references, each of which is incorporated by reference in its entirety for all purposes: Callahan M, Beard C: Ptosis, 4th ed. Birmingham: Aesculapius, 1990; Dresner SC: Further modifications of the Müller&#39;s muscle conjunctival resection procedure. Ophthalmic Plast Reconstr Surg 1991;7:114-122; Dresner SC: Minimal ptosis management. In: Kikkawa DO, ed. Aesthetic Ophthalmic Plastic Surgery. Philadelphia: Lippincott-Raven, 1997:151-162; Older II: Ptosis repair and blepharoplasty in the adult. Ophthalmic Surg 1995;4:304-308; and Crawford IS: Repair of ptosis using frontalis muscle and fascia lata: a 20 year review. Ophthalmic Surg 1977; 8:31-40. The amount of levator function present generally determines which surgical procedure will be adopted. For minimal ptosis Mülllerectomy or Fasanella-Servat procedures have been used. Levator aponeurotic surgical repair has been used for patients with involutional changes. Frontalis suspension and Whitnall&#39;s sling have been used for more severe cases of ptosis. In the frontalis suspension operation, the eyelid is suspended from the frontalis so that the eyelid is opened when the patient lifts the eyebrow using the frontalis muscle. Tendon tissue from the patient&#39;s leg or biocompatible synthetic materials are also used. While this procedure allows the patient to raise the eyebrow to open the eyelid and therefore see from the eye, it suffers from a number of drawbacks. The patient must adapt to the uncommon, tiring and uncomfortable movement of raising the eyebrow to raise the eyelid. Furthermore, the extent to which the patient is able to raise the eyelid varies from procedure to procedure. Essentially, the procedure restores some eyelid function but that function is not natural. This procedure is also a cosmetic failure because of the requirement for the patient to raise his or her eyebrow. U.S. Pat. No. 5,522,889 to Baker, et al entitled “Apparatus and method for restoring eyelid function,” teaches an apparatus to restore eyelid function in a patient unable to voluntarily raise an eyelid. The apparatus includes a spiral torsion spring and pulley arrangement mounted in a housing that is implanted in fixed positions in the superior portion of the orbit of the eye. A wire connects the pulley to the eyelid. A spiral torsion spring provides the necessary spring force in tension to overcome the weight of the eyelid and draw the eyelid open. The natural muscles of eye closure are, however, sufficiently strong to overcome the spring tension thereby paying out wire from the pulley and closing the eye so as to provide normal blinking function. A position setting gear allows the biasing force of the spring to be selectively reduced sufficiently to allow the eye to remain closed for sleep or at other desired times. SUMMARY OF THE INVENTION The present invention provides a novel surgical method and apparatus to restore eyelid function to patients suffering from conditions such as ptosis. The present invention teaches a surgical procedure and method overcoming the above-described limitations and disadvantages of the prior art to restore essentially normal eyelid function in patients unable to raise their lid due to a congenital or acquired condition. The present invention provides an implantable eye drop activated artificial muscle network to take over the function of the levator palpebrae superioris muscle in a relatively simple manner to be readily surgically implantable in the eye yet providing reliable operation over an extended period. The present invention provides a surgical method, artificial muscles and eye drops for correcting ptosis in a patient whereby the eyelid is moved in a natural motion so as to restore substantially natural eyelid function. Additional advantages and other novel features of the invention will be set forth in part in the description that follows and in part will become apparent to those skilled in the art upon examination of the following or may be learned with the practice of the invention. Other advantages and features of the present invention will become apparent to those skilled in the art from the following description wherein there is shown and described a preferred embodiment of the present invention, simply by way of illustration of one of the modes best suited to carry out the invention. Accordingly, the drawings and descriptions will be regarded as not as restrictive and only illustrative in nature. BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings incorporated in and forming a part of the specification, illustrate several aspects of the present invention and together with the description serves to explain the principles of the invention. In the drawing: FIG. 1 is a view of a patient suffering from ptosis in one eye. FIGS. 2( a, b ) are schematic zoomed-in cross sectional views through the eye of a patient showing the relative implanted position of PAN artificial muscle fibers of the present invention for restoring eyelid function in an expanded relaxed (a) or a contracted (b) configuration; FIGS. 3( a, b ) are schematic zoomed-out cross sectional views through the eye of a patient showing the relative implanted position of PAN artificial muscle fibers of the present invention for restoring eyelid function in an expanded relaxed (a) or a contracted (b) configuration; FIG. 4 is a schematic front view of the eye of a patient showing the tarsal glands and relative implanted position of PAN artificial muscle fibers of the present invention for restoring eyelid function; Reference will now be made in detail to the present preferred embodiment of the invention, an example of which is illustrated in the accompanying drawing. DETAILED DESCRIPTION OF THE INVENTION The current disclosure provides apparatus and methods for correcting ptosis. FIG. 1 shows a patient suffering from ptosis in one eye. As seen in FIG. 1, a typical ptosis manifests itself in the form of upper eyelid droop under normal conditions. Typically, a surgical approach can be implemented to correct the droopy eyelid. According to the present disclosure, an artificial muscle can be surgically implanted into the eyelid. The implant can then be biased in either an open or closed position with application of different biasing means. The biasing means may take the form of different chemical solutions, for example, in the form of eyedrops having different chemical properties such as different acidities. As will be described in greater detail below, according to one embodiment, biocompatible fibrous contractile and expansive ionic polymeric artificial muscles can be surgically implanted and sutured under the superior palpebral conjunctiva and suture anchored to the tissues of the upper superior fornix. The artificial muscles can be sutured in a serpentine network or grid fashion to form a two-dimensional sheet suture anchored under the superior palpebral conjunctiva tissues in a serpentine and parallel configuration with respect to tarsal (meibomian) glands of the eyelid. The muscles can be further suture anchored to the tissues of upper fornix in the eyelid. The sutured artificial muscles can then be manipulated by the patient to transform between a contracted (open) position and a relaxed (closed) position via the use of eyedrops of different acidities. The use of artificial muscles as implants in the human body are described in U.S. Pat. Nos. 6,168,575, 6,464,655, 6,511,508, 6,589,198, and 6,682,500, each of which is incorporated by reference in its entirety for all purposes. Ionic polymeric artificial muscles and, in particular, contractile artificial muscles are described in the following references, each of which is incorporated by reference in its entirety for all purposes: M. Shahinpoor, K. J. Kim and Mehran Mojarrad, “Ionic Polymeric Conductor Composite Artificial Muscles,” ERI/AMRI Press, Albuquerque, N.Mex., 2nd. Edition, (2005); M. Shahinpoor, “Ionic Polymer Conductor Composite Materials as Distributed Nanosensors, Nanoactuators and Artificial Muscles—A Review”, Proceedings of the Second World Congress On Biomimetics and Artificial Muscle (Biomimetics and Nano-Bio 2004), Dec. 5-8, 2004, Albuquerque Convention Center, Albuquerque, N.Mex., USA, (2004); M. Shahinpoor, “Ionic Polymer Conductor Composites As Distributed Nanosensors, Nanoactuators and Artificial Muscles—A Review of Recent Findings”, Proceeding of The International Conference on Advanced Materials and Nanotechnology, AMN-1, The MacDiarmid Institute for Advanced Materials and Nanotechnology, 9-11 Feb. 2003, Wellington, New Zealand, pp. 14-22, (2003); M. Shahinpoor, et al, “Soft Actuators and Artificial Muscles”, US Patent Office, U.S. Pat. No. 6,109,852, Issued Aug. 29, (2000); M. Shahinpoor, et al, “Ionic Polymer Sensors and Actuators”, US Patent Office, U.S. Pat. No. 6,475,639, Issued Nov. 5, 2002; M. Shahinpoor, “Artificial Muscles”, Chapter in Encyclopedia of Biomaterials and Biomedical Engineering, edited by G. Wnek and G. Bowlin, Marcel Dekker Publishers, New York, N.Y., (2004); M. Shahinpoor and A. Guran, “Ionic Polymer-Conductor Composites (IPCC) as Biomimetic Sensors, Actuators and Artificial Muscles, SELECTED TOPICS IN STRUCTRONICS AND MECHATRONIC SYSTEMS, Editors: A. Belyaev and A. Guran, pp. 417-436, World Scientific Publishers, London, (2003); M. Shahinpoor, “Ionic Polymer-Conductor Composites As Biomimetic Sensors, Robotic Actuators and Artificial Muscles-A Review”, Electrochimica Acta, Vol. 48, No. 14-16, pp. 2343-2353, (2003); K. J. Kim and M. Shahinpoor, “Application of Polyelectrolytes in Ionic Polymeric Sensors, Actuators, and Artificial Muscles”, Review Chapter in Handbook of Polyelectrolytes and their Applications, edited by S. K. Tripathy, J. Kumar and H. S. Nalwa, vol. 3; Applications of Polyelectrolytes and Theoretical Models, American Scientific Publishers, Stevenson Ranch, Calif., USA (2002); K. J. Kim and M. Shahinpoor, “A Novel Method of Manufacturing Three-Dimensional Ionic Polymer-Metal Composites (IPMC&#39;s) Biomimetic Sensors, Actuators and Artificial Muscle,”, Polymer, Vol. 43/3, pp.797-802 (2002); M. Shahinpoor and K. J. Kim, “Novel Ionic Polymer-Metal Composites Equipped with Physically-Loaded Particulate Electrode As Biomimetic Sensors, Actuators and Artificial Muscles”, Actuators and Sensors A, Physical, 96 (2/3) A, 3163, pp. 125-132, (2002); M. Shahinpoor, Y. Bar-Cohen, J. Simpson and J. Smith, “Ionic Polymer-Metal Composites (IPMC&#39;s) As Biomimetic Sensors, Actuators and Artificial Muscles—A Review”, Smart Materials &amp; Structures Int. Journal, vol. 7, pp. R15-R30, (1998); and M. Shahinpoor, M., “Active Polyelectrolyte Gels As Electrically-Controllable Artificial Muscles and Intelligent Network Structure”, Book Chapter in Structronic Systems, Part II, edited by: H. S. Tzou, A. Guran, U. Gabbert, J. Tani and E. Breitbach, World Scientific Publishers, London, pp. 31-85, (1998). As stated above, the present disclosure provides artificial muscles that can be implanted into the eyelid and sutured to as to allow for the correction of ptosis. The implanted artificial muscles can be formed from a highly resilient biocompatible silicone or other medical grade rubber and/or may include or be formed from a fibrous polyacrylonitrile (PAN) hydrogel. PAN hydrogel is a multiblock copolymer that is formed from a combination of hard blocks (nitrile group) and soft blocks (hydrophilic groups), the proportion of which can be changed to modify the physical properties. PAN hydrogels have good biocompatibility and low toxicity. Compared with other hydrogels, PAN hydrogels have high tear strength. The FDA approved a hydrogel form of PAN in 2002 for a number of medical uses including cervical dilation during childbirth and gastro esophageal reflux disease (GERD). Contractile fibrous gels of PAN artificial muscles are described in U.S. Pat. No. 5,389,222 as well as the references cited below, each of which is incorporated by reference in its entirety for all purposes: K. J. Kim, K. Choe, R. Samathan, J. Nam, M. Shahinpoor and J. Adams, “Toward Nanobiomimetic Muscles: Polyacrylonitrile Nanofibers”, Proceeding of SPIE 11th Annual International Symposium on Smart Structures and Materials, 14-18 Mar., 2004, San Diego, Calif., SPIE Publication No. 5385-62, pp.33-43, (2004); K. J. Kim, J. Caligiuri and M. Shahinpoor, “Contraction/Elongation Behaviour of Cation-Modified Polyacrylonitrile Fibers”, Proceeding of SPIE 10th Annual International Symposium on Smart Structures and Materials, 2-6 Mar., 2003, San Diego, Calif., SPIE Publication No. 5051-23, pp. 207-213, (2003); K. J. Kim and M. Shahinpoor, “Electrical Activation of Contractile Polyacrylonitrile (PAN)-Conductor Composite Fiber Bundles As Artificial Muscles”, Proceedings of the First World Congress On Biomimetics and Artificial Muscle (Biomimetics 2002), Dec. 9-11, 2002, Albuquerque Convention Center, Albuquerque, N.Mex., USA, (2002); M. Shahinpoor and M. Ahghar, “Modeling of Electrochemical Deformation in Poly-acrylonitrile (PAN) Artificial Muscles”, Proceedings of the First World Congress On Biomimetics and Artificial Muscle (Biomimetics 2002), Dec. 9-11, 2002, Albuquerque Convention Center, Albuquerque, N. Mex., USA, (2002); K. J. Kim, J. Caligiuri, K. Choi, M. Shahinpoor, I. D. Norris, B. R. Mattes “Polyacrylonitrile Nanofibers as Artificial Nano-Muscles”, Proceedings of the First World Congress On Biomimetics and Artificial Muscle (Biomimetics 2002), Dec. 9-11, 2002, Albuquerque Convention Center, Albuquerque, N.Mex., USA, (2002); M. Shahinpoor, K. J. Kim, L. O. Sillerud, I. D. Norris, B. R. Mattes, “Electroactive Polyacrylonitrile Nanofibers as Artificial Nanomuscles”, Proceeding of SPIE 9th Annual International Symposium on Smart Structures and Materials, San Diego, Calif., SPIE Publication No. 4695-42, (March, 2002); M. Shahinpoor, K. J. Kim, and H. B. Schreyer, “Artificial Sarcomere and Muscle Made with Conductive Polyacrylonitrile (C-PAN) Fiber Bundles”, Proceedings of SPIE 7th International Symposium on Smart Structures and Materials, Newport Beach, Calif., Vol. 3687, pp. 243-251 (March, 2000); H. B. Schreyer, N. Gebhart, K. J. Kim, and M. Shahinpoor, “Electric Activation of Artificial Muscles Containing Polyacrylonitrile Gel Fibers”, Biomacromolecules, Vol. 1, No. 4, pp. 642-647, (2000); H. Brett Schreyer, Mohsen Shahinpoor, Kwang Kim, “Electrical activation of PAN Artificial Muscles”, Proc. SPIE Smart Materials and Structures Conference, Mar. 1-5, 1999, New Port Beach, Calif., Publication No. SPIE 3669-19, pp. 192-198. (1999); M. Shahinpoor, “Active Polyelectrolyte Gels as Electrically-Controllable Artificial Muscles and Intelligent Network Structures”, Book Chapter, in Active Structures, Devices and Systems, edited by H. S. Tzou, G. L. Anderson and M. C. Natori, World Science Publishing, Lexington, Ky., (1997); Salehpoor, K., Shahinpoor, M. and Mojarrad, M., “Some Experimental Results On The Dynamic Performance of PAN Muscles”, Smart Materials Technologies, SPIE Publication No. Vol. 3040, pp. 169-173, (1997); K. Salehpoor, M. Shahinpoor and M. Mojarrad, “Electrically Controllable Artificial PAN Muscles”, Proc. SPIE 1996 North American Conference on Smart Structures and Materials, Feb. 27-29, 1996, San Diego, Calif., vol. 2716, paper no. 07, (1996); See also, Y. Pierre Gobin, MD, Fernando Vinuela, MD, Harry V. Vinters, MD, Cheng Ji, MD and Kira Chow, MD, “Embolization with Radiopaque Microbeads of Polyacrylonitrile Hydrogel: Evaluation in Swine”, Radiology, vol. 214, pp. 113-119, January (2000) and, Va Stoy, “New type of hydrogel for controlled drug delivery”, J Biomater Appl., vol. 3, No. 4, pp. 552-604, April (1989) According to one embodiment, the implanted artificial muscles can transform between multiple positions or operating modes. Transformation can be activated via a biasing means. According to this embodiment, the biasing means allows the patient to selectively shift eyelid position from a first position or operating mode to a second position or operating mode. For example, the first position or operating condition can be a contracted, or open, position. In this position, it may be desirable that the strength of contraction of the artificial muscles by the biasing means is less than the strength of the contractile fibers in the eye, so as to allow for blinking. The second position or operating condition may be a relaxed, or closed position. The biasing means can be, for example, a chemical solution. Moreover, the chemical solution may be provided in eye drop form. Accordingly, the implanted artificial muscles can be chemically-activated. For example, the implanted artificial muscles may be formed from a chemically activated fibrous PAN hydrogel. As a non-limiting example, the artificial muscle fibers can be formed from a pH active material such as a pH active hydrogel form of fibrous PAN artificial muscles. The pH active artificial muscles can be implanted such that different muscular behavior can be achieved when the muscles are stimulated with solutions containing different pH levels. For example, according to one method, upon using a mildly acidic solution such as an eye drop having a pH of around 4, an example of which is Ciloxan® ophthalmic solution (Alcon labs, Fort Worth, TX), the PAN muscles contract and draw the eyelid open by providing the necessary resilient contraction force in tension to overcome the weight of the eyelid and draw the eyelid open. The natural muscles of eye closure or the orbicularis muscle are, however, sufficiently strong to overcome the artificial muscle tension and resiliency thereby stretching the highly resilient and compliant PAN muscle fibers and closing the eye so as to provide normal blinking function. A mildly alkaline solution, such as an eye drop with a pH of around 8, an example of which is a Timolol® solution, will, in turn, enable the PAN muscle fibers to expand and relax to allow the eye to remain closed for sleep or at other desired times. The biocompatible contractile PAN artificial muscles generally have two distinct resilience coefficients or moduli of elasticity in contraction and in expansion, respectively, upon pH activation as reported in the following papers: M. Shahinpoor, K. J. Kim, and H. B. Schreyer, “Artificial Sarcomere and Muscle Made with Conductive Polyacrylonitrile (C-PAN) Fiber Bundles”, Proceedings of SPIE 7th International Symposium on Smart Structures and Materials, Newport Beach, Calif., Vol. 3687, pp. 243-251 (March, 2000); H. B. Schreyer, N. Gebhart, K. J. Kim, and M. Shahinpoor, “Electric Activation of Artificial Muscles Containing Polyacrylonitrile Gel Fibers”, Biomacromolecules, Vol. 1, No.4, pp. 642-647, (2000); M. Shahinpoor, K. J. Kim and Mehran Mojarrad, “Ionic Polymeric Conductor Composite Artificial Muscles,” ERI/AMRI Press, Albuquerque, N.Mex., 2nd. Edition, (2005); each of which is incorporated herein by reference. FIGS. 2( a, b ) are schematic zoomed-in cross sectional views through the eye of a patient showing the relative implanted position of PAN artificial muscle fibers 19 for restoring eyelid function in an expanded relaxed (a) or a contracted (b) configuration. In FIGS. 2( a ) and 2 ( b ) the eye anatomy is denoted by 1 where the upper eyelid 2 and the lower eyelid 3 are such that the upper eyelid 2 is drooping in the presence of levator palpebrae superioris muscle 13, the orbital septums 14 and 14 ′, the superior tarsal (Muller&#39;s) muscle 15, the superior conjunctival fornix 16, the inferior conjunctival fornix 16 ′, the orbicularis oculi muscles 17 and 17 ′, the superior tarsus 18, the inferior tarsus 18 ′, the sclera 9, the bulbar conjunctiva 10, the superior palpebral conjunctiva 12, the zonules 11, the cornea 4, the crystalline lens 8, the anterior chamber 5, the iris 7, and the posterior chamber 6. As shown, the implanted contractile artificial muscle fibers 19 are sutured under the superior palpebral conjunctiva in a serpentine and parallel configuration with respect to tarsal (meibomian) glands of the eyelid and suture anchored to the tissues of upper superior fornix 21 and the lower edge of the eyelid 20. The serpentine-sutured contractile artificial muscle fiber diameter can be about 100 microns in overall diameter and can be composed of strands of about 10 microns in diameter. In FIG. 2( a ) the contractile muscle fibers 19 are in a relaxed and expanded state while in FIG. 2( b ) the contractile fibrous muscles are in a contracted state. According to one method, the sutured implanted eye drop activated PAN resilient contractile artificial muscles can be such that they provide a resilient force in tension in the range of 0 to a few 10&#39;s of grams. Accordingly, the sutured implanted artificial muscles upon activation with one or more chemical stimulants can have a range of deformation of about 20 mm between contraction and expansion. As stated before, the chemical stimulants can take the form of solutions have different chemical properties such as differing pH levels. For example, an acidic eye drop solution can be used to penetrate the superior palpebral conjunctiva and contract the polymeric artificial muscle fibers. The contractile fibers are so resilient that they allow the blinking or closing of the eyelid by the powerful orbicularis oculi muscles. In contrast, an alkaline eye drop can penetrate through the superior palpebral conjunctiva and cause the polymeric muscles to relax and expand so that the eye lid can easily close with minimum effort when the patient desires to close his or her eyes for extended periods of time. The effort required for eye lid closure is particularly reduced when the sympathetic Muller&#39;s muscles relax right before a person falls asleep. According to one embodiment, the acidic solution may have a pH level of around 4 and the alkaline solution may have a pH level of around 8. It will be understood that additional chemical solutions including, but not limited to, chemical solutions having different pH levels can be used and that the present disclosure encompasses solutions having the appropriate chemical property required to stimulate movement of the artificial muscles in an appropriate and/or desired way. Turning now to FIGS. 3( a ) and 3 ( b ), the eye anatomy is denoted by 1 where the upper eyelid 2 and the lower eyelid 3 are such that the upper eyelid 2 is drooping in the presence of the sclera 9, the bulbar conjunctiva 10, the superior palpebral conjunctiva 12, the zonules 11, the cornea 4, the crystalline lens 8, the anterior chamber 5, the iris 7, the posterior chamber 6. As shown, the implanted contractile artificial muscle fibers 19 are sutured under the superior palpebral conjunctiva in a serpentine and parallel configuration with respect to tarsal (meibomian) glands of the eyelid and suture anchored to the tissues of upper superior fornix 21 and the lower edge of the eyelid 20. In FIG. 3( a ) the contractile muscle fibers 19 are in a relaxed and expanded state while in FIG. 3( b ) the contractile fibrous muscles are in a contracted state. Again, the contractile fibers are so resilient that they allow the blinking of the eyelid or closing of the eyelid by the orbicularis oculi muscles. FIG. 4 is a schematic front view of the eye of a patient 1 showing the upper eyelid 2 and the lower eyelid 3 are such that the upper eyelid 2 is flipped backward to see the upper and lower tarsal gland 3 and 3 ′ in the presence of the sclera 9, the limbus 10, the cornea 11 and the iris 7. As shown, the implanted contractile artificial muscle fibers 19 are sutured under the superior palpebral conjunctiva in a serpentine and parallel configuration with respect to tarsal (meibomian) glands of the eyelid and suture anchored to the tissues of upper superior fornix 21 and the lower edge of the eyelid 20. The foregoing description of a preferred embodiment of the invention has been presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the invention to the precise form disclosed. Obvious modifications or variations are possible in light of the above teachings. The embodiment was chosen and described to provide an illustration of the principles of the invention and its practical application to thereby enable one of ordinary skill in the art to utilize the invention in various embodiments and with various modifications as are suited to the particular use contemplated. All such modifications and variations are within the scope of the invention as determined by the appended claims when interpreted in accordance with the breadth to which they are fairly, legally and equitably entitled. Unless specifically stated to the contrary, all references cited in the present disclosure are incorporated by reference in their entirety for all purposes.

Summary: A surgical procedure is described for the restoration of eyelid function in individuals suffering from ptosis or upper eyelid droop syndrome that makes a patient unable to voluntarily fully raise an eyelid. The surgical procedure includes implantation and suturing of eye drop (pH) activated and actuated fibrous contractile and expansive artificial muscles such as pH active hydrogels of polyacrylonitrile (PAN) artificial muscles that are surgically implanted and sutured under the superior palpebral conjunctiva in a serpentine parallel configuration with respect to the tarsal (meibomian) glands of the upper eyelid and anchored to the tissues of superior fornix.

big_patent

en

Summarize: CROSS-REFERENCES TO RELATED APPLICATIONS This application is related to the application entitled &#34;COFFEE BREWER&#34; of which Alan M. King is the inventor, Ser. No. 599,712, filed Apr. 12, 1984, now U.S. Pat. No. 4,632,023. BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates in general to coffee or other beverage brewers, and in particular to an improvement in beverage brewing machines wherein the filter element can be repetitively used and can be removed from the brewing chamber so that the coffee grounds can be removed therefrom and then the filter element can be returned to the brewing chamber. 2. Description of the Prior Art The present invention is an improvement in beverage brewing machines such as disclosed in U.S. Pat. No. 3,565,641 which issued on Feb. 23, 1971 wherein the inventor is Alan M. King. This patent discloses a beverage brewing vending machine for brewing a single cup of coffee or other beverage which has a brewing chamber which receives hot water and beverage material. The chamber has a floor through which gas and liquid can pass but is provided with a filter which prevents the beverage material from passing therethrough. The apparatus disclosed in U.S. Pat. No. 3,565,641 has a lower chamber which is of substantially the same cross-sectional size as the first chamber located below the first chamber in which a piston is mounted. The piston can be moved upwardly toward the first chamber which forces air through the floor of tee first chamber into the first chamber so as to agitate and brew the hot water and beverage material mixture to produce the beverage. The piston then moves away from the floor of the top chamber and draws the beverage through the floor and filter and then dispenses it through a suitable spout. In the apparatus of U.S. Pat. No. 3,565,641, the filter material is not reused but a supply of filter material is supplied to the brewing chamber for each individual cup of coffee and then is discarded. SUMMARY OF THE INVENTION It is an object of the present invention to provide a beverage brewing machine which allows the same filter material to be reused repetitively and provides for the upper chamber being raised from the lower brewing chamber during which time the filter material in the brewing chamber and the residue from the prior cup of beverage passes out of the brewing chamber and is scraped from the filter material after which the filter material is then returned to the brewing chamber and the upper brewing chamber then rests against the lower brewing chamber so as to provide a seal. Another object of the invention is to provide a lever operated grid hold down mechanism which during the portion of the brewing operation when the piston is moving upwards to the top of the cylinder, engages and holds the filter down, and is removed from engagement with the filter material when the piston reaches top dead center and remains at its maximum distance from the said filter material during the balance of the brewing cycle. It is yet another object of the invention to provide an improved brewing machine which allows the filter material to be reused but which provides the option to remove the reusable filter material and use instead a non-reusable paper filter material. Another object of the invention is to provide a brewing machine wherein a large portion of the brewing cycle is used for drawing coffee through the filter and agitating the beverage and where there is very limited time of the cycle for raising the upper chamber and holding it up and removing the grounds and filter material out of the brewing chamber and returning the filter material to the brewing chamber and lowering the upper chamber and the hold down grid. Another object is to provide a positively driven filter material web which is returned to the brewing chamber. Other objects, features and advantages of the invention will be readily apparent from the following description of certain preferred embodiments thereof taken in conjunction with the accompanying drawings although variations and modifications may be effected without departing from the spirit and scope of the novel concepts of the disclosure. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a view illustrating the beverage brewing machine of the invention; FIG. 2 is a top view of the beverage brewing machine; FIG. 4 is a partially sectional view of the beverage machine; FIG. 5 is a partially cut-away side view of the machine; FIG. 6 is a sectional view taken on line VI--VI in FIG. 5 and illustrates the mechanism for controlling the movement of the upper brewing chamber and the filter material; FIG. 7 is a side view of the brew hold down drive for the beverage machine; FIG. 8 is a partially cut-away sectional view taken on line VIII--VIII in FIG. 5 illustrating the pull down lever; FIG. 9 illustrates the filter medium drive; FIG. 10 illustrates a modification of the invention; and FIG. 11 illustrates the modification of FIG. 10. DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 illustrates the structure of the present machine which comprises a frame member 130 with a coffee dispenser 121 mounted thereon with an opening 123a and which has a suitable check mechanism such as well known to those skilled in the art which upon demand deposits coffee into an upper brewing chamber 11 which cooperates with a lower brewing chamber 25. Hot water is dispensed from a reservoir 123 which has a suitable heater and a spout 124 for providing one cup of hot water into the funnel 17 of the top brewing chamber 11 where it is agitated by the action of a piston 26 mounted in the lower chamber 25 to brew coffee. After the coffee has been brewed, it is drawn through a filter and passes out openings 33 to a spout 101 to a cup 102 for use. FIGS. 2 through 9 disclose the mechanism of the invention. As shown in FIGS. 4 and 6, the top of the lower brewing chamber 25 supports the filter material 36. The filter material 36 may be a screening fabric type such as polyester monofilament number HD7-75 or HD7-63 which is available from TETKO Inc., 420 Saw Mill River Rd., Elmsford, N.Y. 10523. The lower chamber 25 is cylindrical-shaped and a piston 26 is mounted therein and is connected by wrist pin 28 to a piston rod 37 which is connected by a pin 29 to a crank arm 31 mounted on drive shaft 32 which is driven in a single direction to actuate the machine by a suitable driving motor 150. The upper brewing chamber 11 may be square, rectangular or round in shape but in the illustrated example, is generally square and has the funnel 17 which has an extension 18 that is mounted to the top of the upper chamber 11 and has a downward funnel portion 19 which terminates at an opening 21 for supplying the hot water and beverage material as, for example, coffee grounds to the brewing chambers. Above the top of the lower chamber 25, the flexible filter material 36 is mounted as shown, for example, in FIGS. 4 and 6. The filter material 36 is sealed in the brewing chamber by a lower gasket 35 mounted in the upper portion of the lower chamber 25 and a second gasket 40 mounted in the lower edge of the upper chamber 11. Molded into gasket 35 is a screen material to prevent grounds from accidently entering the lower chamber 25. A filter material hold down grid 20 engages the filter material 36 during the brewing cycle and has three longitudinal members 42, 44 and 43 as illustrated in FIG. 2 and five transverse members 41, 46, 47, 48 and 49. Tee hold down grid 20 is connected to a pivotally supported hold down lever 52 by bearings 51 and 51a and brackets 53 attached to the grid hold down member 20 as shown in FIGS. 2 and 4. In the present invention, the upper brewing chamber 11 is opened and unsealed from the lower brewing chamber 25 to allow the strip of filter material 36 to be withdrawn from the brewing chamber so as to remove the grounds 101 from the previous cup of coffee from tee machine. This is generally illustrated in FIG. 9 wherein the strip 36 of filter material has one end which passes over a roller 10 to a reel 103 mounted on the frame of the machine with a shaft 104. The shaft 104 carries a gear 106 which is in mesh with an idler gear 105 mounted on shaft 99 which meshes with a segment gear 107 which drives tee reel 103. The segment gear 107 is mounted on shaft 108 which carries a sprocket 109 and cams 111 and 118. Cam 111 engages a cam follower 112 mounted on an arm 113 which is pivotally supported as illustrated in FIG. 3. The end 116 of arm 113 is bent as shown and is received in an opening 114 in frame 58 which serves as a pivot point for moving the filter hold-down member 20. The arm 113 is pivotally connected to an arm 117 which has its upper end 120 connected to member 122, as shown in FIG. 5. Member 122 has a pin 251 which is received in a recess formed between uprights 252 and 253 formed on member 135 which is pivotally supported on shafts 59 and 59a of the frame 58. Member 122 has an upper surface against which the end 262 of rocker arm 261 bears. Rocker arm 261 is pivotally supported on shaft 54 which has a flat portion as shown in FIG. 4. Lever arm 52 is carried on shaft 54 and has a free end with a pin 51 which engages brackets 700 and 701 that are attached to hold down member 20. Thus, when push rod 117 is pulled down by cam 111 the hold down member 20 is held down. Cam 118 engages a cam follower 119 mounted on a lever 142 which is pivotally mounted with a pin 144 as shown in FIG. 3. Lever 142 carries a rod 143 and when cam follower 119 rises on cam 118, filter material 36 is positively pulled by rod 143. As shown in FIG. 9 the filter web 36 is connected to the reel 103 by pin 131 such that when gear 106 is driven by idler gear 105 which is in turn driven by segment gear 107 the reel 103 pulls the filter material 36 out of the brewing chamber such that the ground residue 101 from the previous brewed cup is scraped off of the filter material 36 by a scraper 132 which has an end 133 that removes the residue 101 and the residue falls into a container, not shown. The other end of the filter material 36 passes over roller 100 and the filter material 36 passes around the rod 143 and has its second end attached to a pin 136 as best shown in FIG. 9. After the segment gear 107 has driven the filter material 36 so that the residue 101 has been scraped from the filter material, the segment gear which rotates in a counterclockwise direction relative to FIGS. 3 and 9 disengages the gear 105 which releases the reel 103 so that the rod 143 will pull the filter material through the brewing chamber so that the rod 143 takes the position shown in dotted line in FIG. 9. The upper brewing chamber is mounted on bifurcated arms 137a and 137b and 138a and 138b which are attached to a pivoted member 135 as shown in FIG. 5. A pair of stub shafts 139 and 141 are connected to the upper brewing chamber 11 and are pivotally mounted in the end of arms 137 and 138 as shown in FIG. 9. The stub shafts 139 and 141 are formed with flat spots as illustrated in FIG. 5 and are received in slots 501 formed in the arms 137 and 138. The rear end of member 135 is pivotally connected to the frame by shafts 59 and 59a as illustrated in FIG. 2 which extend through frame members 58a and 58b connected to the frame 58 of the machine. A spring 201 is mounted on a stub shaft 59c and has ends 202 and 203 so as to bias the member 135 and the arms 137 and 138 upwardly. A shaft 164 passes through member 135a as illustrated in FIG. 4 and a washer assembly 172 is mounted on the extensions 135a and nut 170 locks the upper end of shaft 164 to the member 135. A center portion 171 of member 135 carries the projection 135a as illustrated in FIGS. 3 and 4. As shown in FIG. 7, the shaft 164 has a hollow enlarged portion 163 at its lower end and carries a cam follower 161 mounted on shaft 162 that is connected to the portion 163. Cam followed 161 engages a cam 154 mounted on shaft 32 which as illustrated in FIG. 5 is driven by the motor 150 through a coupling member 151 and pin 600. A sprocket 300 is also mounted on shaft 32 as illustrated in FIG. 7 and FIG. 5 and drives a chain belt 301 which drives a sprocket 109 mounted on shaft 108 illustrated in FIG. 3. As illustrated in FIG. 3, sprocket 109 is mounted on shaft 108 and carries cams 111 and 118 as well as segment gear 107 which engages gear 105 to drive the filter material 36 on to reel 103. Cam 111 engages cam follower 112 in pivoted arm 113 so as to drive the arm 117 to actuate the filter medium hold-down lever as described. Cam 118 engages cam follower 119 in pivoted lever 142 and rod 143 returns the filter material 36 from reel 103. The shaft 32 also drives the crank arm 31 which is connected by a pivot pin 29 to piston rod 37 which is pivotally connected by pivot 28 to the piston 26 as illustrated in FIGS. 6 and 8, for example. In operation, when the dispenser machine is energized, the motor 150 illustrated in FIG. 5 will drive the drive shaft 32 in one direction and coffee will be dispensed from the holder 121 and hot water from the reservoir 123 into the upper brewing chamber 11. At this time, the grid hold down 20 engages the filter material 36 through the action of roller 112 on cam 111 and the rest of the linkage to hold i down and the piston 26 moves upwardly in the cylinder 25 forcing air through the filter 36 to agitate the hot water and coffee grounds in the upper chamber 11 so as to brew coffee rapidly and efficiently. During this time, the upper brewing chamber 11 is firmly held against the lower brewing chamber, so the gaskets 40 and 35 seal the brewing chamber. When the piston has passed top dead center, the filter hold down 20 is released by the action of roller 112 on cam 111 which allows the arm 117 to rise. A spring 254 is mounted on pin 251 so as to bias the member 122 upwardly. As 117 is attached to arm 122, this permits arm 261 to rise as it is biased by spring 260. Thus hold down 20 is allowed to also rise on cam 111 acting through roller 112 and the linkage associated with same before all the grounds settle on it. Then the piston moves downwardly in the cylinder 25 which sucks the brewed coffee through the filter material 36 into the lower brewing chamber on to the top of the piston 26. As piston 26 continues downwardly in the cylinder 25, it passes the slots 33 which allow the brewed coffee to pass therethrough. The push rod 164 moves upward due to the action of the cam 154 on the cam 154 on the cam follower roller 161 which causes the upper brewing chamber 11 to be raised due to the biasing spring 201. When the piston passes bottom dead center (pouring out the brewed coffee) and begins its upward motion, the grid 20 is lowered to hold down the filter. It is held in this position until the piston reaches top dead center. When said grid is raised, it remains there for the balance of the brew cycle. The purpose of this is to have the grid away from the filter when the patty is formed. With the grid hold down 20 in the up position and the upper brewing chamber 11 in the upper position the segment gear 107 engages the gear 105 which engages gear 106 to move the filter material 36 and used coffee grounds 101 out of the brewing chamber past the scraper 132 and the grounds are removed and fall into a suitable receptacle. Then the segment gear 107 passes beyond the gear 105 and the rod 143 returns the filter material 36 to the brewing chamber and the upper brewing chamber 11 and the hold down grid 20 are returned to the down position ready for the next cycle with the piston 26 in the lower position. FIGS. 10 and 11 illustrate a modification wherein the reuseable filter material 36 can be replaced with a roll of filter paper and wherein the machine of the invention can be modified to use the roll of filter paper such that each portion of the filter paper is only used one time. The filter paper 36a passes between the roller 103 which is driven by the gear 106 and a roller 602 supported by shaft 603. The roller 602 is supported on arm 604 which has a portion 605 which engages a cam 606 rotating on shaft 108 When roller 602 is out of engagement with reel 103 the paper 36a is not driven. It is seen that this invention provides an improved brewing machine wherein a large portion of the cycle is used for brewing the beverage such as coffee and there is a very limited time allowed for raising the upper brewing chamber and the hold down grid and removing the grounds and filter material out of the brewing chamber to clean it and then return the filter to the brewing chamber and again lower the upper brewing chamber and the hold down grid. The present invention comprises a substantially improvement in brewing machines of the prior art. Although the invention has been described with respect to preferred embodiments, it is not to be so limited as changes and modifications can be made which are within the full intended scope of the invention as defined by the appended claims.

Summary: A beverage brewing machine which has upper and lower brewing chambers which are movable relative to each other so as to clamp a strip-shaped filter therebetween during the brewing process and the chambers can be separated so as to allow the filter strip to be removed from the brewing chamber and the beverage residual wiped therefrom after which the filter strip is returned to the brewing chambers which are then resealed for the next cycle. A hold down grid engages the top surface of the filter to hold down as a piston moves in the lower chamber upwardly to force air through the filter to agitate the beverage. When the piston reaches top dead center, the hold down grid is lifted. When the piston passes the opening the brewed coffee is poured.

big_patent

en

Summarize: BACKGROUND OF THE INVENTION This invention relates to planters in general and more particularly relates to planter apparatus and a method of plant care which assures that the planting medium is well irrigated without subjecting the plant roots to over irrigation and/or insufficient irrigation. The instant invention concerns a type of planter in which there is an inner container for soil, an outer container wherein the inner container is disposed, a space constituting a basin between bottom walls of the inner and outer containers and the bottom wall of the inner container being apertured to permit water to drain into the basin from the inner container. Prior art constructions of this type are disclosed in U.S. Pat. No. 4,192,097 issued Mar. 11, 1980 to W. J. Smith for Horticultural Improvements and U.S. Pat. No. 4,335,540 issued June 22, 1982 to R. P. Allen for Combined Plant Container and Watering Device. A serious problem encountered when utilizing prior art planters of this type is that even though the soil appears to be moist and watering occurs at regular relatively short intervals, because water flows naturally along paths of least resistance (so-called &#34;short circuit effect&#34;), certain portions of the soil are not moistened. That is, each time watering takes place the water takes the same favored paths in draining from top to bottom and these paths &#34;short circuit&#34; or bypass certain areas of the soil. When the prior art has sought to overcome the &#34;short circuit effect&#34; the results have been expensive structures and/or procedures that are cumbersome, and more often than not resulted in overwatering or failed to eliminate sour soil and/or excessive salt buildup so that leaf tips and margins turned yellow or brown, plants rotted at or above soil level, leaves dropped, leaves and/or petals became spotted or completely discolored, leaves curled or otherwise distorted, and/or leaves or stems wilted. BRIEF DESCRIPTION OF THE INVENTION To overcome these problems of the prior art, utilizing a double container or pot arrangement, the instant invention provides an irrigating procedure in which the soil in the inner pot is submerged in water for a selected period of time, say between five and fifteen minutes, during which time water also fills the space or basin between the bottom walls of the inner and outer containers and extends well up into the inner container, approximately to the top surface of the soil. After the five to fifteen minute interval, more water is added until all of the soil is below water. Thereafter, water is drained from the basin while water drains from the inner container through the perforated bottom thereof. In a preferred embodiment of the instant invention the water level for the inner container is monitored and a siphon is utilized to drain water from the basin at a controlled rate so that most of the water is permitted to drain from the inner container and the plant roots are buried in damp soil rather than in soil that is submerged in water. In accordance with the instant invention, water can be forced upward through the perforated bottom of the inner container to drain from the inner container over the top edge thereof or through one or more side apertures that are located slightly above the soil level. For the embodiment in which a siphon is utilized to drain water from the basin, the normal outlet end of the siphon hose is raised above the upper soil level during the upper desalting procedure. After this procedure takes place the siphon outlet is directed downward once again and water is evacuated from the basin below the inner container. Accordingly, the primary object of the instant invention is to provide a reliable irrigating method consisting of simple procedures. Another object is to provide simplified equipment for carrying out the irrigating method taught by the instant invention. Still another object is to provide equipment of this type that is simple to operate. A further object is to provide relatively inexpensive planter apparatus which permits simplified plant care. BRIEF DESCRIPTION OF THE DRAWINGS These objects as well as other objects of this invention shall become readily apparent after reading the following description of the accompanying drawings in which: FIG. 1 is a perspective of planter apparatus constructed in accordance with teachings of the instant invention. FIG. 2 is a perspective of a modified form of the outer container which comprises a portion of the apparatus illustrated in FIG. 1. FIG. 3 is a vertical section taken through the center of simplified apparatus for carrying out major portions of the irrigating method according to teachings of the instant invention. FIG. 4 is a perspective looking at the top and two corners of another simplified apparatus for carrying out major portions of the irrigating method according to teachings of the instant invention. FIG. 5 is a side elevation of still another simplified version for carrying out major portions of the irrigating method according to teachings of the instant invention. DETAILED DESCRIPTION OF THE INVENTION Now referring to the Figures and more particularly to FIG. 1 which illustrates planter apparatus 10 that includes outer container 11, inner container 12 disposed within outer container 11 and a draining means in the form of siphon tube 14. The sidewalls of both containers 11 and 12 are circular and slightly tapered. As will hereinafter be seen, while a relatively tight fit between the outer wall of inner container 12 and the inner wall of outer container 11 will reduce the amount of liquid to be drained, such relatively tight fit is not required in order to carry out the instant invention. Containers 11, 12 are proportioned so that the perforated bottom wall 16 of inner container 12 is spaced from bottom wall 17 of outer container 11 to form upper and lower boundaries of catch basin 18 which receives liquid that drains from inner container 12 through apertures 19 in bottom wall 16 thereof. Cleaning of basin 18 is facilitated by removing inner container 12 from outer container 11. Formed integrally with outer container 11, extending the full height thereof and disposed on the outside thereof, is spout 20 having internal central wall 21 that divides spout 20 into side-by-side vertical channels 22, 23. The lower end of channel 22 is in communication with basin 18 by means of aperture 39 and, for a reason to be hereinafter explained, near its upper end channel 23 is provided with drain aperture 26 that is aligned with an aperture of similar size in the sidewall of inner container 12, which aperture 26 is disposed slightly above the upper level of the soil in inner container 12. Legs or standoffs 29 project downwardly from bottom wall 17 of outer container 11 and rest against the bottom wall 31 of pan 30 so as to maintain a space between bottom 17 of outer container 11 and the inside surface of bottom wall 31 for pan 30. Sidewall 32 of pan 30 is provided with spout 33 which makes it convenient to pour liquid from pan 30. Siphon tube 14 includes inlet portion 41 that is disposed vertically within channel 22, outlet portion 42 that is disposed vertically within channel 23 and curved sighting portion 43 that connects the upper ends of tube portions 41 and 42. Sighting portion 43 is disposed outside of spout 20 and is positioned slightly below drain aperture 26. The free or lower end 46 of inlet portion 41 is disposed at the bottom of channel 22 and constitutes the inlet for siphon 14, while the free bottom end 47 of outlet portion 42 constitutes the outlet 47 for siphon 42, which outlet 47 is disposed below spout 20 in that the lower end of outlet portion 42 extends through oversized aperture 48 at the bottom of channel 23. Thus, liquid discharges from tube 14 directly into pan 30. Liquid that accumulates in pan 30 may be discarded or may be poured into inner container 12 during a subsequent watering operation. As an alternative to accumulating liquid in pan 30 tube 14 may be directed to discharge directly into a drain. By having a substantial portion 43 of tube 14 disposed outside of spout 20 removal and replacement of tube is facilitated. To utilize planter apparatus 10 inner container 12 is filled with soil or other planting medium 51 (FIG. 3) wherein roots 52 of plant 53 are buried. The height of soil 51 is slightly below the upper edge of outer container 11, and for a reason to be hereinafter explained, curved section 43 of siphon tube 42 extends slightly above the soil level. The number and size of the perforations 19 are sufficient to permit water or other plant nourishing liquid to drain relatively freely into basin 18 from the interior of inner container 12, yet wall 16 is strong enough to support soil 51 even when it is saturated. The irrigating operation is commenced with siphon hose 42 in the position of FIG. 1. Water, either plain or fortified with nutrients, is poured into channel 22 through the enlarged top thereof and/or into the open upper end of inner container 12 until soil 51 within container 12 is soaked with water as indicated by the level of water in channel 22 and inlet portion 41 of siphon tube 14. The latter is transparent, at least in the vicinity of its curved portion 43. When water reaches a desired height within inlet portion 41, this height being somewhat below curved portion 43, the addition of water is halted for a predetermined interval, typically five to fifteen minutes. During this period the water distributes itself so that basin 18 is filled and most of the soil 51, starting upward from the bottom 16 of inner container 12, is immersed in water. This condition will be indicated by a slight drop in the water level sighted in siphon tube inlet portion 41, which level will stabilize. Thereafter, more water is added slowly through the top of channel 22 or at the upper end of inner container 12. This causes the level of water within inlet portion 41 to rise, finally reaching a level in curved portion 43 where siphoning action commences and water flows from exit 47 into pan 30 at a rate which is controlled largely by the diameter of siphon tube 14. This flow continues until essentially all of the water is evacuated from catch basin 18. If water is permitted to remain standing within pan 30 air in the vicinity of planter apparatus 10 will remain moist. Water collected in pan 30 may be poured into other planters or may be discarded as desired. To wash salts from the upper portion of soil 51, tube outlet portion 42 is removed from channel 23 and extended upward so that outlet 47 is substantially above inner container 12. Thereafter water is introduced into channel 22 through its upper end with this water flows upward through perforations 19 and upward through soil 51 and exiting inner container 12 through an aperture that is aligned with aperture 26, flowing downward, exiting channel 23 through aperture 48 at the bottom thereof and entering pan 30. Outer container 11 with spout 20 thereon may be replaced by outer container 71 of FIG. 2 which includes vertical channel-like protrusion or spout 72 extending for the full height thereof. Basin 73, of rectangular cross-section, extends diametrically across bottom 74 of outer container 71 and projects therebelow. The upper or interior surface of bottom wall 74 slopes gradually from the periphery thereof toward basin 73. Spout 72 is intended to replace channel 22. Inlet portion 41 of siphon 14 is intended to extend through spout 72 and outlet portion 42 is to be disposed outside of spout 72 with connecting hose portion 43 extending through side aperture 75 with only slight clearance. The latter is located in wall 63 of spout 72 near the upper end thereof. Outer container 71 near the upper edge thereof is provided with a plurality of drain holes 26. It is intended that when inner container 12 is inserted in outer container 71 upper drain holes 26 will be aligned with upper drain holes (not shown) in inner container 12 and that none of these upper drain holes will be aligned with spout 72 so that water which discharges through upper holes 26 flows along the outside of outer container 71 into pan 30. Outer container 71 provides three sidewalls 61, 62, 63 which define spout 72. The fourth sidewall or closure for spout 72 is provided by the curved sidewall of inner container 12 and this sidewall is closely fitted to the inner surface of the sidewall for outer container 71. The aligned upper drain holes in containers 12 and 71 near the tops thereof is utilized for a salts washing operation of the type described for the emobodiments of FIG. 1, with draining salts completely bypassing spout 72. The apparatus illustrated in FIGS. 1 and 2 may be modified by eliminating siphon tube 14 and using other means for gauging the height of water within inner container 12. For container 71 of FIG. 2 spout 72 may be constructed either in whole or part of transparent material so that the height of water within inner container 12 can be gauged by observing the height of water within spout 72. With such an arrangement a pluggable or valve controlled drain hole will be provided at the bottom of spout 72 for draining water from inner container 12 and from catch basin 73. FIG. 3 illustrates a simplified version of apparatus for carrying out the plant irrigating method according to the instant invention. That is, planter apparatus 80 of FIG. 3 includes inner container 81 disposed within outer container 82, with the perforated bottom wall of container 81 being spaced from bottom wall 84 of container 82 to form catch basin 85 therebetween. Outer container 82 is disposed within pan 30 at the center thereof being supported on spacing means comprising a plurality of legs 86. Opening 88 in the sidewall of outer container 83 at the bottom thereof provides a passage for draining water from catch basin 85 into pan 30. Normally, opening 88 is closed by removable plug 89 which is removed when water is to be drained from basin 85 and also when reverse flushing is to take place. That is, plug 89 may be removed and a hose (not shown) connected at opening 88 so that water may be introduced into basin 85 and forced upward through soil in and then out of aligned upper side openings 91a, 91b, of the respective outer and inner containers 82, 81 to urge salts from the upper portion of soil 51. Such hose may be transparent and be used for sighting (gauging) of the water level in container 81. Another alternativve is to replace plug 89 by a valve (not shown). For the embodiment of FIG. 3 with plug 89 in place filling of outer container 81 takes place through the top thereof and should proceed slowly enough to permit water to drain into basin 85 without an excessive amount of water rising above the top of soil 51. This filling operation should be halted when water flows through top openings 91 as fast as it is being supplied to inner container 81. In the embodiment of FIG. 4 rectangular container 101 is divided by vertical partition 102 into relatively large soil containing chamber 103 and relatively narrow water level sighting chamber 104. Water catch basin 105 extends from and communicates with the bottom of chamber 104, and is below bottom 106 of chamber 103. Apertures 107 near the bottom of partition 102 permit liquid to drain from chamber 103 into basin 105. Other regions of partition 102 may be provided with additional apertures to facilitate draining into chamber 104 and to permit root aeration. Removable drain plug 108 is provided at one end of basin 105 near the bottom thereof. Extension 110 projects downward from bottom 106 of container 101 along the edge thereof remote from basin 105. The end of basin 105 remote from plug 108 is provided with short leg 109 and the bottom of basin 105 slopes so that with extension 110 and leg 109 resting on a horizontal support surface water in basin 105 drains toward plug 108. Apertures 181 near the upper end of chamber 103 are provided for back flushing. In the embodiment of FIG. 5 container 120 is provided with internal ledge 121 near the bottom thereof which supports perforated partition 132 horizontally to divide container 120 into a relatively deep soil containing section 122 and a relatively shallow basin 123 disposed below section 122. Input end 126 of siphon tube 125 is connected to basin 123 at aperture 124 and outlet end 127 of tube 125 extends below bottom 129 of basin 123. Tube 125 is bent at its midregion 128 which is positioned near but below upper edge 131 of container 120. Mark 133 on the entrance side of midregion 128 is near the upper level of soil within section 122 and is used for sighting the height of liquid in container 120. Apertures 183 near the upper edge of container 120 are provided for back flushing. It should now be apparent to those skilled in the art that the containers and pans herein described may be molded plastic elements and that formations at the bottom and/or side of an outer container may be formed integrally therewith. Further, a spout and/or siphon tube may be constructed of transparent material and then be used for sighting (gauging) of water level in the inner container. Indicia to indicate operating levels may be marked on the gauging elements required. Although the present invention has been described in connection with a plurality of preferred embodiments thereof, many other variations and modifications will now become apparent to those skilled in the art. It is preferred, therefore, that the present invention be limited not by the specific disclosure herein, but only by the appended claims.

Summary: A method of caring for plants is carried out by utilizing a planter that includes a soil container having openings at the bottom thereof for communication to a basin. The container is filled with liquid to a level at which substantially all of the soil is submerged in the liquid and remains so for a selected period of time. Then an aperture at the bottom of the basic is opened and the liquid in the container drains through the openings at the bottom of the latter into the basin and exiting therefrom through the aperture. In another embodiment a transparent siphon is utilized to remove liquid from the basin and is also utilized as a sighting means so that a user is aware that liquid in the container has reached a desired level. Salts that often accumulate at the soil surface are run off by having liquid enter the container through the openings at the bottom thereof and run out of the container above the soil line.

big_patent

en

Summarize: Hostess Brands Inc. is nearing a deal to sell its Twinkie brand and other cakes to private-equity firms Apollo Global Management LLC and C. Dean Metropoulos & Co. for more than $400 million, said people familiar with the discussions. Hostess is nearing a deal to sell its Twinkie brand and other cakes to private-equity firms Apollo Global Management and C. Dean Metropoulos for more than $400 million. Rachel Feintzeig reports on The News Hub. Photo: Reuters. A deal would serve as the opening bid in a coming bankruptcy-court auction for the assets, which include Dolly Madison and other brands. The so-called stalking horse bid could be topped by other suitors at the auction, in which case Apollo and Metropoulos would likely be entitled to what's known as a breakup fee. One person cautioned a deal might not be disclosed until later this week, but the parties were putting finishing touches on the agreement Tuesday. The deal, which includes several factories, caps Hostess's efforts to sell off popular cake and bread brands as it winds down operations this year. Hostess, based in Irving, Texas, in November announced it was shutting down and selling of its 30 or so brands and 36 plants, a move expected to result in the loss of more than 18,000 jobs. Hostess moved to liquidate after it failed to reach a deal on cost cuts with its second-largest union, representing thousands of bakers. Hurst Capital, a fledgling private-equity firm run by 26-year-old twins, also vied for Hostess's cake brands, but appears to have fallen short of securing the stalking horse bid. Apollo, the giant buyout shop co-founded by Leon Black, and Metropoulos, owner of the Pabst Blue Ribbon beer brand, have been looking to partner on a deal for some time. In recent years they've explored buyouts including of Sara Lee Corp. They emerged as the front-runners to snap up Hostess's cake brands and negotiations between them and the company picked up steam in recent days, leading to the expected deal, people familiar with discussions said. Metropoulos is known for investing in well-known food brands, including Vlasic pickles, Lender's bagels and Mrs. Butterworth's syrups. Meanwhile, Apollo has been a player in food retail. The firm owns a majority stake in Sprouts Farmers Market, which has about 150 natural-food stores in the western U.S., and last year sold Smart & Final Inc., a chain of about 250 warehouse stores, to fellow private-equity firm Ares Management LLC. in deal valued at $975 million. Flowers Foods Inc., the Thomasville, Ga.-based maker of Tastykakes and Nature's Own breads, is offering up to $360 million in cash for five major Hostess bread brands—including Wonder and Nature's Pride—along with 20 plants and 38 depots. In addition, the baking company is offering $30 million for Hostess's Beefsteak rye brand. A judge Friday cleared Hostess to place those bread assets on the auction block on Feb. 28. Hostess Monday debuted two more sale deals—one for a group of bread brands including Sweetheart, Eddy's, Standish Farms and Grandma Emilie's, and one for the Drake's brand, which made treats like Devil Dogs, Ring Dings and Yodels before Hostess began liquidating in bankruptcy. United States Bakery Inc., a Portland, Ore., company also known as Franz Family Bakery, is offering $28.85 million for the bread brands plus four bakeries, 14 depots and equipment. McKee Food Corp., the Collegedale, Tenn.-based maker of Little Debbie snack cakes, has submitted a $27.5 million bid for the Drake's brand and certain equipment. —Ryan Dezember contributed to this article. Write to Mike Spector at mike.spector@wsj.com and Rachel Feintzeig at rachel.feintzeig@dowjones.com A version of this article appeared January 30, 2013, on page B4 in the U.S. edition of The Wall Street Journal, with the headline: Hostess Nears Twinkies Deal. Hostess has picked the maker of Little Debbie as the lead bidder for its Drake's cakes. According to a filing in U.S. bankruptcy court, McKee Foods has offered $27.5 million in cash for the cake brands, which include Devil Dogs, Funny Bones and Yodels. The fate of Twinkies and other Hostess cakes are still being negotiated with other bidders. Hostess also said United States Bakery agreed to pay $28.9 million for its remained bread brands, which include Sweetheart, Eddy's, Standish Farms and Grandma Emilie's. That offer includes four bakeries, 14 depots and equipment. Earlier this month, Hostess picked Flowers Foods, which makes Tastykake and Nature's Own and Bunny bread, as the lead bidder for six of its major bread brands, including Wonder. The "stalking horse" bid by McKee Foods would set the floor for an auction process that lets competitors make better offers. A judge would have to approve the final sale, which Hostess said is scheduled to close no later than May 30. McKee's bid includes some equipment but not the Drake's bakery in Wayne, N.J. A spokesman for Hostess, Tom Becker, said the company continues "to market all remaining assets." McKee Foods, based in Collegedale, Tenn., makes a variety of snack cakes under the Little Debbie banner that compete with Hostess cakes at a lower price. For example, its Cloud Cakes resemble Twinkies and its Devil Cremes resemble Devil Dogs. A representative for McKee Foods, Mike Gloekler, said the company didn't plan to scrap any brands as a result of the deal. "Our intent is to produce like products as they are since they have different packaging and formulae," Gloekler said in a statement. He said McKee hoped to make Drake's products at its plant in Stuarts Draft, Va. since Drake's cakes are best known in the northeast region. Hostess has said in court previously that it needed to move quickly in selling off its brands to capitalize on the outpouring of nostalgia and media coverage prompted by its demise. The company repeated the sentiment in its court filing Monday, noting that there is no advertising or marketing for Drake's brands, which also include Ring Dings, Sunny Doodles and Yankee Doodles. "The longer Drake's products stay off the shelves, the more likely it is that consumers will begin to use competitors' products," the filing said. McKee generates about $1.1 billion in sales a year, with its Little Debbie cake division accounting for $800 million of that, according to the company. In recent years, McKee has seen its sales remain flat or fall as eating habits have changed. Hostess Brands Inc., based in Irving, Texas, has been plagued by even greater problems. The company announced in November that it was shutting down its business and selling its breads and snack cakes. Its demise came after years of management turmoil and turnover, with workers saying the company failed to invest its brands. Hostess filed for its second Chapter 11 bankruptcy in less than a decade this January, citing costs associated with its unionized workforce. After declaring that it was going out of business, Hostess had solicited bids for its brands by a Dec. 10 deadline. The company said in its filing Monday that it had received one bid for "substantially all" of its assets. But Hostess said the bid was not as valuable as the combined total for of the bids it received for individual brands. In addition, Hostess said the bidder that made the offer for conducted "very limited diligence." ___ Follow Candice Choi at www.twitter.com/candicechoi Getty Images The private-equity firm that owns Pabst Blue Ribbon beer is planning to bid on the Hostess cake brands division – which includes Ding Dongs, Ho Hos, Drake’s Devil Dogs, and…Twinkies. Could a PBR-like revival be in the snack cake’s future? PBR and Twinkies may not seem similar. One is a cheap-but-popular beer, especially beloved in hipster urban neighborhoods. The other a once-pervasive childhood staple with a bit-role in the Hostess bankruptcy proceedings. But both are iconic legacy brands that were once very much in demand. PBR has been revived. Twinkies ain’t doing so well. (MORE: 10 Tips for Getting the Most Out of College Aid) For PBR, 2001 was the low point. After winning awards more than a century earlier (we’re talking 1880s) and seeing production peak in 1977, the brand began a slow decline into obscurity. Over a span of about 25 years, Pabst went from making 18 million barrels to 1 million. That was right before the company discovered a subset of Americans who were really into Pabst. Namely, hipsters. They appreciated that PBR didn’t market itself the way that the Bud Lights of the world, with their ubiquitous ads, were doing it. So PBR pounced, as it were, targeting areas in New York and Portland with offbeat ways of spreading the brand — it sponsored bike polo tournaments or supported local bands – while being careful not to overdo it. It appeared as if the brand wasn’t trying, a guiding ethos of their target audience. The revival of the PBR brand spurred private equity firm C. Dean Metropoulos & Co. to buy Pabst in 2010. Metropoulos has a history of buying flagging brands, including Chef Boyardee and Bumble Bee Tuna. The firm is now eyeing Twinkies. Back in November, it was reported that Metropoulos was interested, but Bloomberg News is now saying the firm is officially in the middle of the bidding process. (MORE: The Next Gun Fight) The question is what the Metropoulos would do with the brand and whether it would try a PBR-style revival. Clearly, Twinkies’ traditional target audience would be different: kids and the parents who fill their sack lunches. But if we could give Metropolous two words of advice, it’d be this: hipster moms. The same college-aged women who helped spur PBR’s revival in the early 2000s now likely have children. Start by pairing Twinkies alongside PBR at select bars: $1 a can, 50 cents a Twinkie. Being a little tipsy will make it much easier for them to ingest the snack cake. This will not only immediately boost Twinkie consumption, but those buzzed moms may even get emotional while eating the yellow snack, triggering thoughts from their childhood and inspiring them to give their kids a Twinkie every now and then. Good luck with the bid, C. Dean Metropoulos & Co. And you’re welcome.

Summary: Goodbye Hostess Twinkies, hello... Apollo Twinkies? Or maybe PBR Twinkies? The Wall Street Journal reports that two private-equity companies-Apollo Global Management and C. Dean Metropoulos-are close to a deal worth more than $400 million to buy the brand from bankrupt Hostess. The bid would serve as the one to beat in an upcoming bankruptcy-court auction, and it now seems unlikely any other suitor would do so. Metropoulos, Time notes, already owns Pabst Blue Ribbon. "Both are iconic legacy brands that were once very much in demand," writes Josh Sanburn. "PBR has been revived. Twinkies ain't doing so well." Meanwhile, the AP reports that the maker of Little Debbie (McKee Foods) is the lead bidder for the Drake's brand, which includes Devil Dogs and Ring Dings. The offer is $27.5 million.

multi_news

en

Summarize: CROSS REFERENCE TO RELATED APPLICATION [0001] This application is a divisional application of U.S. application Ser. No. 10/345,635, filed Jan. 16, 2003, which is hereby incorporated herein in its entirety by reference. BACKGROUND OF THE INVENTION [0002] This invention relates to surgical needles for tissue ablation, and more particularly, to surgical needles that are for ablation of uterine fibroids. [0003] Approximately 20 to 40 percent of women have uterine fibroids (lieomyomata). In the United States, fibroids result in approximately 175,000 hysterectomies and 20,000 myomectomies each year. Fibroids are well-defined, non-cancerous tumors that arise from the smooth muscle layer of the uterus. Approximately 25% of women suffer fibroid related symptoms, including menorrhagia (prolonged or heavy menstrual bleeding), pelvic pressure or pain, and reproductive dysfunction. [0004] The most common treatments for fibroids include hysterectomy, abdominal myomectomy, laparoscopic myomectomy, hysteroscopic myomectomy, laparoscopy-directed needle mylosis, laparoscopy-directed needle cryomyolysis, high-intensity focused ultrasound ablation of fibroids, and uterine artery embolization. Hysterectomy is a major surgical procedure and carries with it the usual risk of surgery, such as hemorrhaging, lesions, complications, pain, and prolonged recovery. The majority of myomectomies are performed abdominally, wherein a surgeon creates an abdominal incision through which individual fibroids are removed. Abdominal myomectomy and laparoscopic myomectomy, like a hysterectomy, carries the usual risk of surgery. [0005] Radio Frequency (RF) myolysis and thermal tissue ablation are two promising methods for treating fibroids. RF myolysis is a technique in which a RF probe is inserted into a fibroid or the surrounding tissue and then RF energy is applied to the tip of the probe. The tissue surrounding the tip is heated by the RF energy causing necrosis within the tissue. Thermal tissue ablation is a technique that is performed with a cryoablation probe. The cryoablation probe destroys the fibroid tissue by freezing it. [0006] Current methods incorporating RF or cryoablation techniques require direct visualization of the needle tip or electronic imaging. Normally, under direct visualization techniques an endoscope is inserted into the uterus to position the needle. Direct visualization is often problematic because of the difficulties involved in simultaneously manipulating the endoscope and needle. Typically, when electronic imaging is used, the position of the needle is visualized with a hysteroscope or with an external abdominal ultrasound. Hysteroscopy allows direct visualization of the uterine cavity by inserting a small camera on the end of a long tube directly into the uterus through the vagina and cervix. Similar to an endoscope, a hysteroscope must be simultaneously manipulated with the needle, and thus is problematic. Monitoring the probe&#39;s position with current ultrasound techniques has a number of drawbacks. For example, a clinician using ultrasound imaging from an external source will have difficulty in distinguishing the uterine tissue from the surrounding organs and precisely locating the needle. [0007] U.S. Pat. No. 5,979,453 to Savage et al. describes a myolysis needle that requires laparoscopic surgery. In laparoscopic surgery the needle must be placed through the uterine serosa into or near the fibroid. As a result, uterine adhesions often form that may cause chronic pain, infertility, and bowel obstruction. Additionally, during laparoscopic surgery the surgeon cannot visualize the tissue below the surface and must blindly place the needle, as a result placement may be sub-optimal. [0008] U.S. Pat. No. 6,146,378 to Mikus et al. discloses a needle placement guide having an endoscope that is inserted into the uterus through the vagina. Using the endoscope, the surgeon positions the endoscopic guide in the correct orientation to the targeted fibroid. After positioning the guide, the endoscope is removed from within the guide and an ablation device is inserted into the guide for subsequent operation on the fibroid. The needle guide suffers from several disadvantages. There is the risk that the needle guide could shift during removal of the endoscope and insertion of the ablation device, resulting in sub-optimal performance. The needle cannot be relocated during the ablation procedure and the endoscope must be reinserted whenever it is necessary to reposition the needle guide. Reinserting and removing the endoscope and ablation device every time the needle must be repositioned increases the time and expense of the surgery. [0009] U.S. Pat. No. 6,379,348 to Onik describes a mylolysis needle that is a combination of a cryosurgical and electrosurgical instrument for tissue ablation. The cryo/electro needle is not easily visualized when in use and requires the use of a dilator to create an access channel in the tissue area where the needle is to be inserted. Similar to laparscopic surgery, placement of the cryo/electro needle is done blindly and may not result in optimal performance. [0010] Thus, a need exists to provide a medical needle system and method that can provide accurate and reliable targeting of fibroid tumors. It is also desirable to provide a needle that has a safety system that would shut-off electrical current to the needle if the uterine wall is punctured. BRIEF SUMMARY OF THE INVENTION [0011] The invention provides a medical needle for transvaginal ultrasound directed reduction of fibroids. The medical needle is adapted for use in conjunction with a transvaginal ultrasound probe. The ultrasound probe has an attached needle guide through which the needle is inserted. The needle has an outer tubular member having an inner surface, a distal end, and a proximal end. The distal end of the outer member is made of an echogenic material so that the tip of the needle has heightened visibility on an ultrasound screen. Located at the distal end is an active electrode that is in communication with a radiofrequency source. An insulating sheath surrounds the entire outer member except for a section that is near the active electrode at the distal end. [0012] The needle has a return electrode that is optionally located on the outer member near the active electrode or on an outer tissue surface of a patient. Optionally, the needle may have a temperature sensor that is located near the active electrode. Typically, the distal end will either be a sharpened pointed tip or a beveled tip that defines an opening in the distal end. [0013] In a preferred embodiment, the needle has a safety device that will turn off power to the active electrode if the tip of the needle should penetrate a patient&#39;s uterine wall. In the embodiment possessing a beveled tip, an inner cylindrical member having a forward end and blunt rear end is disposed within the outer member. The inner member has a cylindrical outer section that is electrically conductive and a section that is not electrically conductive. Disposed on the inner surface of the outer member is a second electrically conductive surface and a third electrically conductive surface that are not in communication with one another. The second surface is in communication with the RF power source and the third surface is in communication with active electrode. [0014] A spring is attached to the forward end of the inner member and the blunt rear end extends outwardly beyond the beveled tip. When pressure is applied to the blunt rear end the spring is compressed and the exposed blunt rear end slides backwardly into the outer member. As the inner tubular member slides into the outer member the electrically conductive surface comes in contact with both the second and third surface so that current passes through the surfaces and RF energy is supplied to the active electrode. [0015] In a second embodiment having a safety device, the inner tubular member does not have a conductive surface and there are no second and third conductive surfaces. Rather, a switch is located at the proximal end of the outer member. When pressure is applied to the blunt rear end of the inner member, the inner member slides back into the outer member and thereby closes the switch. When in the closed position, the switch sends a signal to the RF source and RF energy is applied to the active electrode. [0016] In a third embodiment, the needle has an outer member, an inner surface, an echogenic distal end, and a proximal end. As in the first embodiment, the echogenic material results in the tip of the needle having a heightened visibility. Within the outer member is a cryogen tube that extends longitudinally from the proximal end to the distal end. Surrounding a section of the outer member from the proximal end to near the distal end is a cryo-insulation sheath. The distal end is in communication with a cryogen supply so that the distal end can be in cryogenic contact with fibroids. [0017] The length of the needle in all embodiments is typically from about 25 to 50 centimeters, and somewhat more typically between 30 to 40 centimeters. The diameter of the needle in all embodiments is typically from about 12 to 18 gauge, and somewhat more typically from about 16 to 18 gauge. Normally, the needle has a handle at the proximal end that allows the user to easily grip and manipulate the needle. [0018] The invention also includes a method for the electric surgery of fibroids using a transvaginal ultrasound directed echogenic needle. The method comprises the steps of providing a transvaginal ultrasound probe having a transducer and attached needle guide; providing an echogenic needle as described above; inserting the probe into a patient&#39;s uterus; inserting the needle into the uterus through the attached needle guide; sensing the location of the needle and fibroid using ultrasound imaging; guiding and positioning the needle on the surface of a fibroid using ultrasound imaging; and passing a controlled amount of RF energy through the fibroid. The method optionally includes the steps of monitoring tissue temperature, penetrating the surface of the fibroid with the distal end of the needle, and the step of turning off power to the active electrode if the distal end pierces the uterine wall. [0019] The invention additionally includes the method for the cryoablation of fibroids in the uterus using a transvaginal ultrasound directed echogenic needle. The method includes the steps of providing a transvaginal ultrasound probe having a transducer and an attached needle guide; providing a cryoablation echogenic needle as described above; inserting the probe into the uterus; inserting the echogenic needle into the uterus through the attached needle guide; sensing the location of the needle and fibroid using ultrasound imaging; guiding and positioning the needle on the surface of a fibroid using ultrasound imaging; delivering a controlled amount of cryogenic supply to the distal end of the needle while it in contact with the surface of the fibroid. The method optionally includes the step of penetrating the fibroid with the distal end of the needle before or after delivering a controlled amount of cryogenic supply. BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING(S) [0020] Having thus described the invention in general terms, reference will now be made to the accompanying drawings, which are not necessarily drawn to scale, and wherein: [0021] FIG. 1 is a side view of a transvaginal ultrasound probe having an attached echogenic needle that has been inserted into a uterus; [0022] FIG. 2 is a perspective view of an ultrasound monitor displaying an echogenic needle that has been inserted into a uterus; [0023] FIG. 3 is a side view of a radio frequency echogenic needle system for use with a transvaginal ultrasound probe; [0024] FIG. 4 is a sectional side view of the needle shown in FIG. 2 ; [0025] FIG. 5 is a sectional side view of a radio frequency echogenic needle having a “shut-off” mechanism; [0026] FIG. 6 is a sectional side view of a radio frequency echogenic needle having a “shut-off” mechanism and a noninsulated segment that is an active electrode; [0027] FIG. 7 is a sectional side view of a radio frequency echogenic needle having a “shut-off” mechanism and an active electrode disposed proximal to the distal end; [0028] FIG. 8 is a sectional side view of a radio frequency echogenic needle having a “shut-off” mechanism and an active electrode disposed in the inner member; [0029] FIG. 9 is a sectional side view of a radio frequency echogenic needle having a switch “shut-off” mechanism; [0030] FIG. 10 is a sectional side view of a cryogenic ablation echogenic needle; and [0031] FIG. 11 is a side view of a radio frequency echogenic needle having a return electrode attached to a patient&#39;s thigh. DETAILED DESCRIPTION OF THE INVENTION [0032] The present inventions now will be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the invention are shown. Indeed, these inventions may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Like numbers refer to like elements throughout. [0033] Referring more specifically to the drawings, for purposes of illustration, but not of limitation, there is shown in FIG. 1 an embodiment of the invention referred to generally as 10. FIG. 1 illustrates an ultrasound probe 100 having the attached mylosis needle 105 that is inserted into the uterus 15. The ultrasound probe has a transducer located within its tip 30 so that imaging of the uterus and needle are sent to a display for monitoring. Normally, the ultrasound probe 100 includes clamps 35 that attach the needle to the ultrasound probe. Typically, the clamps are made from a metal or plastic material that fits tightly around the probe and has an attached needle guide. The needle guide is typically a narrow or circular opening through which the needle is inserted. Alternatively, the material comprising the clamps is some other hard material that allows the user to manipulate the needle, although not necessarily with equivalent results. The ultrasound probe useful in the invention is any probe that is designed for insertion through the vagina. [0034] As illustrated in FIGS. 1 and 2, the ultrasound probe 100 is inserted into the uterus through the vagina. Once the probe is in place, the needle is inserted through the needle guide and into the uterus. The physician uses ultrasound imagery to locate the position of fibroids 50 and the needle 105 in the uterus. The tip of the needle 160 is directed against a targeted fibroid or its vascular supply and RF energy, cryogenic, or thermal treatment is applied to the fibroid to cause necrosis of the tissue. In this regard, FIG. 2 illustrates an ultrasound monitor 60 that is displaying ultrasound imaging of an echogenic needle 105 that has been inserted into a uterus 15. Normally, the probe sends data to an ultrasound unit 65 that processes the data and then displays the resulting images on the monitor. [0035] In all embodiments, the needle will have an echogenic surface 135 at or near the distal end 120. For example, FIG. 3 shows a bumpy or uneven surface 135 on the outer member. Echogenicity refers to a surface&#39;s ability to reflect incident ultrasound waves back to a sensor. The more a surface reflects waves back to the sensor the greater its image will appear on an ultrasound display. Today, there is a variety of different techniques to increase a surface&#39;s echogenicity, including grooves or recesses, bumps, coatings, indentations, and the like. In the invention, the echogenic tip enhances its visualization and helps the physician to more precisely position the tip. Normally, the distal end of the needle or a segment proximal to the distal end will have an echogenic surface. [0036] Inserting both the ultrasound probe and echogenic needle into the uterus through the vagina is very advantageous. Traditional laparoscopic myomectomy requires that the ablation needle be inserted into the uterus through the abdomen. During this procedure the needle must be inserted through the uterine serosa, which may result in the formation of uterine adhesions. In contrast, the invention provides an apparatus and method of use for fibroid myomectomy that is a minimally invasive surgical procedure. Adhesions are not expected to form with this method because the echogenic needle is inserted through the vagina rather than penetrating the uterine serosa. A second advantage of the invention is precision and accuracy. The echogenic needle has a heightened ultrasonic visibility that allows the physician to accurately locate and position the needle within the uterus. As a result, the surgical procedure is performed more quickly, the needle is easily repositionable by the surgeon, and most importantly the procedure will have a greater beneficial impact for the patient. [0037] With reference to FIGS. 3 through 10, needles that are useful in the current invention are illustrated. The needle has an outer tubular member 115, a proximal end 125, a distal end 120, an insulation sheath 200 surrounding a portion of the outer member, and an echogenic surface 135 near the distal end. [0038] As shown in FIG. 3, a RF needle is broadly designated by reference number 105. The needle 105 includes an active electrode at the distal end 120. Typically, the active electrode is a wire, wire loop, metal surface, or the like. The active electrode is in communication with an electrical connector 140 that is attached to the proximal end 125. The electrical connector 140 is connected to a RF power supply 140 a so that RF current is supplied to the active electrode. The needle 105 is connected to a RF power source 140 a, and optionally to a temperature display (heat readout) 140 b. Normally, the RF source will also include a means for controlling current to the active electrode 140 c. Typically, the RF needles will have a RF insulated sheath 200 that surrounds the outer member 115 and extends from the proximal end 125 to the distal end 120 leaving a segment of the outer member 120 a ( FIGS. 6 and 7 ) that is RF noninsulated. The RF insulation sheath may be made of any material that is suitable to prevent RF energy passing from the outer member to the tissue being treated, such as a heat shrink polyolefin or Teflon®. [0039] The RF needle of the invention delivers either monopolar or bipolar current. With reference to FIGS. 4 through 9, a RF needle having a return electrode 210 is illustrated. The return electrode is connected to the power supply so that current passes through the active electrode into the fibroid tissue and back to the return electrode. Normally, the return electrode is located on the outer shaft 115 about 2 to 20 millimeters from the active electrode. Typically, the return electrode 210 is positioned in close proximity to the active electrode so that RF energy that passes from the active electrode through the fibroid is focused and does not dissipate within the uterus. Alternatively, as illustrated in FIG. 11, the return electrode 210 a is located on an outer surface of the patient, such as the thigh or lower back. In this manner, current passes out of the active electrode 175 through the patient&#39;s tissue, and into the return electrode 210 a. [0040] In FIG. 4, the active electrode 175 is depicted at the distal end 120 within the needle. In this first embodiment, the distal end&#39;s noninsulated outer surface 150 is electrically conductive so that RF energy passes from the active electrode 175 into fibroid tissue. The distal end 120 has a sharpened tip 160 that can penetrate fibroid tissue to deliver RF energy within the fibroid. As shown in FIG. 4, the RF needle optionally has a temperature sensor 185 disposed near the distal end 120. Typically, the temperature sensor will be disposed near the tip of the needle or within the insulation sheath. Normally, the temperature sensor is a thermocouple or thermistor. The sensor provides information that enables the physician to monitor tissue temperature and to adjust the power accordingly. [0041] With reference to FIGS. 5 through 9, reference number 400 broadly designates a RF needle having a RF energy “shut-off” mechanism. The shut-off mechanism turns off RF energy to the active electrode if the tip of the needle 190 penetrates through the uterine wall. Shutting off power to the active electrode serves several useful purposes. It prevents damage to healthy tissue, which would otherwise be coagulated by RF energy and it alerts the physician that the needle has punctured the uterine wall. [0042] In contrast to the first embodiment, RF needle 400 has a sharpened beveled tip 190, an inner cylindrical member 405, and a spring 430 disposed within the outer member 115 at the outer member&#39;s proximal end 125. The inner member 405 is disposed and moveable longitudinally within the outer member 115. As illustrated in FIGS. 5 through 9, the inner member 405 has a forward end 407 and a blunt rear end 425. The forward end 407 is attached to the spring 430 that is connected to the needle&#39;s proximal end 125. In the at rest position, the blunt rear end 425 extends outwardly from the beveled tip 190 and is the first part of the distal end 120 to contact uterine tissue. Applying pressure to the blunt rear end 425 compresses the spring 430, and the inner member 405 slides longitudinally from the distal end 120 towards the proximal end 125. As a result, the blunt rear end 425 retracts into the outer member 115 and the beveled tip 190 contacts the surface of the targeted tissue. [0043] In a first embodiment of RF needle 400, a segment of the inner cylindrical member has a cylindrical conductive surface, and outer member 115 has a second and third conductive surfaces on its inner surface. The second surface is in communication with the RF power supply 140, and the third surface is in communication with the active electrode 175. When in the rest position, the second and third surfaces are not in communication with each other. As pressure is applied to the blunt rear end 425 the inner member 405 retracts into a charged position. When in a charged position, the conductive surfaces 410, 415, and 420 are in communication and RF energy flows from the RF power source to the active electrode. If the distal end 120 punctures the uterine wall pressure against the blunt rear end 425 will be released and the spring 430 will rapidly extend the blunt rear end 425 outwardly. As a result, the conductive surface 410 will move longitudinally away from the second and third surfaces 415, 420 and RF energy supplied to the active electrode is shut-off. The exact position of conductive surfaces 410, 415, and 420 is not critical except that it is necessary that all three surfaces simultaneously communicate with each other when the inner member is in a retracted position. [0044] In this regard, FIG. 6 shows a conductive surface 410 on the inner member 405. The conductive surface 410 is optionally located at the forward end 407 of the inner member 405 or at almost any position along the inner member. The second 420 and third surfaces 415 are located on an inner surface 117 of the outer member 115 so that when the inner member 405 retracts the conductive surfaces 410, 415, and 420 contact each other. When pressure is applied to the blunt rear end 425, the spring 430 compresses and the inner member retracts into the outer member 115. As a result, the conductive surfaces 410, 415, and 420 are in communication with one another and RF energy is delivered to the active electrode 175. [0045] The active electrode is at the distal end 120 or alternatively, the noninsulated surface 120 a of the outer member 115 is the active electrode. In this regard, FIG. 7 illustrates an RF needle having an insulation sheath 435 disposed between the second conductive surface 420 and the outer member 115. RF energy is supplied to the second surface through a current line 440 that is in communication with the electrical connector 140. As shown in FIG. 7, conductive surface 410 on the inner member 405 is in electrical communication with the outer member&#39;s 115 inner surface 117. Typically, the outer member is made from a material, such as stainless steel, that is electrically conductive and suitable for insertion into tissue. When the inner member 405 retracts into the outer member 115 the second surface 420 contacts the conductive surface 410 supplying RF energy to the noninsulated segment 120 a. Optionally, insulation sheath 435 insulates the entire inner surface 117 of the outer member 115 except for segments at the active electrode 120 a and the third conductive surface 415. [0046] In a second embodiment of a needle having a safety mechanism 400, the active electrode is located at the blunt rear end. As shown in FIG. 8, the active electrode 175 is located at the blunt rear end 425 and an electrical connector 425 a extends longitudinally from the conductive surface 410 to the active electrode 175. The outer member 115 has a second conductive surface 420 that is in communication with RF power supply, but rather than having a third surface in communication with the active electrode, the conductive surface 410 on the inner member 405 is in communication with the active electrode 175. When pressure is applied to the blunt rear end 425, the spring 430 compresses and the inner member retracts into the outer member. As a result, the conductive surfaces 410, 415 contact one another and RF current is applied to the active electrode 175. Typically, the electrical connector 425 a is disposed within the inner member 405. [0047] However, the electrical connector 425 a may be disposed between the surface of the inner member and an optional RF insulation sheath that surrounds the inner member. The optional insulation sheath does not surround the conductive surface 410 or the active electrode 175. [0048] In a third embodiment of a RF needle with a safety mechanism 400, the inner member is connected to a switch. With reference to FIG. 9, a needle is shown having an inner member 405 attached to a switch 450. The switch 450 is in communication with a RF power source via line 455. As pressure is applied to the blunt rear end 425 the inner member 405 retracts into the outer member 115 and closes the switch 450. When in the closed position, the switch 450 sends an electrical signal through line 455 to the RF power supply 140 a and RF energy is delivered to the active electrode. The active electrode is located at the distal end and is in communication with the switch, or alternatively, the noninsulated distal end 120 a is the active electrode. [0049] In all the embodiments of a needle having a safety mechanism 400 the inner member 405 is typically made from a material that is non-conductive, such as a plastic. Normally, a non-conductive member will have a conductive material, such as stainless steel, inserted into a surface segment so that the inner member has an electrically conductive surface that will contact the second and third surfaces on the outer member. Somewhat more typically, the inner member is made from a metal such as stainless steel that is surrounded by a RF insulation sheath. The insulation sheath surrounds the inner member except for the conductive surface 410, which is RF non-insulated. [0050] With reference to FIG. 10, a cryoablation needle is broadly illustrated by reference number 500. The cryoablation needle has an echogenic distal end having a sharpened tip 160. The outer member 115 is surrounded by a cryo-insulation sheath 200 a. The insulation sheath 200 a extends longitudinally from the proximal end 125 to the distal end 120 leaving a segment of the outer member 120 a that is cryo-noninsulated. Normally, the sheath will be made of any material that prevents the cryogenic effect from passing through the outer member and into the surrounding tissue. A cryogen supply tube 510 is disposed within the outer member and extends from the proximal end 125 to the distal end 120. A cryogen supply source 520 provides cryogen supply through a cryogen connector 525 to the cryogen supply tube 510. [0051] Typically, cryogenic liquids such as nitrogen, helium and argon are used to produce the cryogenic effect in the targeted tissue. [0052] In all embodiments, it is necessary that the needle is longer than the ultrasound probe and has sufficient length to reach fibroids deep in the uterus. Typically, the length of the needle is about 25 to 50 centimeters, and somewhat more typically about 30 to 40 centimeters. The needle&#39;s diameter is dictated by the ultrasound probe&#39;s attached needle guide. Typically, the diameter of the needle is about 12 to 18 gauge, and somewhat more typically about 16 to 18 gauge. However, the needle is not limited to the above recited dimensions and may be varied depending upon the actual length of the probe and the needle guide&#39;s inner diameter. Typically, the outer member is made of any material that is suitable for insertion into tissue, such as stainless steel. [0053] Optionally, as shown in FIG. 3, the needle will have a handle 130 at its proximal end 125. The handle 130 allows the user to easily manipulate and move the tip of the needle. Ideally, the handle 130 is large enough to be manipulated with the user&#39;s thumb, index finger and middle finger. Normally, the handle is metal, plastic, rubber, or the like. [0054] Many modifications and other embodiments of the inventions set forth herein will come to mind to one skilled in the art to which these inventions pertain having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the inventions are not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation.

Summary: The invention is a transvaginal ultrasound probe having an attached echogenic needle that is useful in the treatment of uterine fibroids. The echogenic needle has an echogenic surface near its tip that allows the physician to visualize its location using ultrasound imaging. In one embodiment, the needle has an active electrode at its distal end. The active electrode supplies radio frequency energy to a fibroids causing necrosis of the targeted fibroid or by destroying the fibroid&#39;s vascular supply. The radio frequency needle preferably has a safety device that shuts-off energy if the needle punctures the uterine wall. In a second embodiment, the needle has a cryogen supply tube and cryogen supply. This embodiment destroys fibroid tissue by freezing it or its vascular supply when the tissue comes in contact with the needle&#39;s frozen distal end. The invention further includes the method of using the ultrasound probe with the attached needle.

big_patent

en

Summarize: Il termine nel contratto di appalto Nel contratto di appalto è usuale inserire un termine entro cui terminare l'opera e è usuale una penale per il ritardo nel completamento dell'opera appaltata. Il problema relativo al termine di adempimento o di completamento dell'opera è dato dal fatto che il contratto di appalto è anche un tipico contratto soggetto a modifiche o variazioni dovute ad eventi interni (volontà del committente) o ad eventi esterni (tempi per autorizzazioni amministrative o dinieghi derivanti da scelte pubbliche ecc.) tutti questi eventi si ripercuotono, inevitabilmente, anche sui termini del contratto di appalto occorre solo valutare quali sono le conseguenze giuridiche. Ovviamente si deve essere in presenza di eventi determinati da terzi (rifiuto o ritardi nel rilascio delle pratiche amministrative) oppure dalle parti (modifica dell'opera di notevoli dimensioni) che per forza incidono sul termine stipulato (impedendo che possa essere rispettato). Se l'esigenza di allungare il termine di consegna dell'opera si manifesta prima della conclusione del contratto grossi problemi non dovrebbero essercene in quanto il contratto di appalto non è perfezionato fino a quando su tutte le clausole del contratto non si è raggiunto il consenso di tutte le parti, quindi, il committente e l'appaltatore potranno contrattare fino a quando non determinano il termine che ritengono più opportuno. Quando, invece, l'esigenza di modificare il termine di consegna dell'opera sorge dopo la stipula del contratto di appalto la situazione diventa più complicata. Infatti, la regola generale è data dal principio che per modificare ogni clausola del contratto è necessario il consenso di tutte le parti, quindi, senza il consenso di entrambi i contrenti nessuna clausola del contratto di appalto può essere modificata. Questo, però, non esclude che il committente e l'appaltatore concordano sulla modifica dell'opera appaltata, ma "si dimenticano" del termine indicato nell'originario contratto, ora, a) se si seguisse la regola generale il termine indicato ab orgine nel contratto resterebbe invariato anche se l'opera ha subito delle modifiche (con necessario ricalcolo del termine); b) oppure, occorre prendere atto che l'originario termine non esiste più (o, quanto meno, non è applicabile), in quanto legato all'originaria struttura / contenuto del contratto.

Summary: Cass. 2.4.2019 n 9152 Quando, nel corso del contratto d'appalto, il committente abbia richiesto all'appaltatore importanti variazioni del progetto - che importino "notevoli modificazioni della natura dell'opera", che determinano una sostituzione del regolamento contrattuale già in essere - il termine di consegna per il ritardo, pattuito nel contratto, viene meno per effetto del mutamento dell'originario piano dei lavori.

fanpage

it

Summarize: Lucan Battison was suspended from a New Zealand Catholic school after he refused to cut his long curly hair but he can keep his hairstyle after winning his fight in the high court today. A student has won his High Court battle to keep his hair long after he was suspended from school because he looked 'unruly'. Lucan Battison, 16, was sent home last month from St. John's College, in Hastings, New Zealand, for not adhering to the rule that states students must keep their hair ‘short, tidy, off their collars and out of their eyes’. Principal Paul Melloy told Lucan he could return once he had his hair cut - but the student argued his naturally curly hair would look 'boofy' if cropped and he was prepared to wear it tied back in order to comply with the school's standards. But New Zealand High Court Judge David Collins ruled that Lucan’s actions did not warrant suspension as his conduct was not harmful to his fellow students. He also said the school had not provided enough clarity on the hair rules to ensure students complied with them. When the case began, the teen’s lawyer, Jol Bates, likened Lucan to Martin Luther King Jr for standing up for his rights. Judge Collins made a point of commenting on Lucan’s character, noting he received a civil bravery award in March for participating in the rescue of two young women who nearly drowned in dangerous ocean conditions. After St John's College student (right) was suspended by principal Paul Melloy the matter was taken to the High Court at Wellington for a judicial review. The teenager is pictured with his lawyer Jol Bates (left) on Monday. He also mentioned that Lucan represented St. John's in rugby and loved attending the school, in part because 'his faith was important to him'. Meanwhile, although the school did not approve of Lucan’s hairstyle, it described him as a ‘nice young man’. Judge Collins said that one of Lucan’s lawyers had tried to provide the school board's disciplinary committee with statements from two hairdressers, one of whom said Lucan's hair was already short and would ‘look like "an untidy afro'' if it was cut shorter,’ but that the committee apparently chose not to accept the testimony. Parents Troy Battison and Tania Doidge said in a statement that their son had never broken the rules because his bun kept his hair off his collar and out of his eyes. 'In 2014, when girls' hair lengths at school aren't questioned, why should the rules be different for boys?' his parents said. 'The criticism we have received as parents has been hurtful and unnecessary.' In a statement issued by the school, Mr Melloy said the school was naturally disappointed about the decision made in Wellington. ‘The Board of Trustees are taking time to consider the judgment made by Justice Collins in terms of its impact, both on our school and on other schools,’ it said. Mr Bates said his client was standing up for his basic human rights, challenging authority and following in the footsteps of Martin Luther King Jr. Mr Bates said Lucan had been sporting the same hairstyle since he became a student at the Catholic college three years ago

Summary: A judge ruled New Zealand schoolboy Lucan Battison does not need to cut his hair before returning to class. The judge also stated that his Catholic high school had been wrong to suspend him for having long locks. The St John's College year 12 student was suspended by principal Paul Melloy last month. The 16-year-old was told by school he could return if he had shorter hair. But Lucan took the matter to the High Court at Wellington instead.

cnn_dailymail

en

Summarize: They pick locks, hang of the edge of multi-storey buildings and dash along underground rail tunnels all in pursuit of the perfect shot to post on social media. Now the lives of Australia’s ‘urban explorers’ are the focus of a documentary that gives an insight into the controversial subculture which police claim is harming young lives, vandalising buildings and disrupting rail networks. In Melbourne, Victoria police say trespassers were caught on the City Loop train tracks more than 1000 times in seven months last year. 'People entering the underground rail network place their lives and the lives of others at risk. We’ve seen deaths and horrific injuries as a result of people entering trains, tampering with equipment and trespassing on rail property,' a Victoria police spokesperson told Daily Mail Australia. Documentary maker Adrian Ortega, 23, spoke with four men involved in ‘urban exploring’ and got four different answers about what it is, how they do it and why they do it. What they all shared was an awareness of the dangerous situations they were putting themselves in. ‘A lot of people have a different stance of what it actually is,’ Adrian told Daily Mail Australia. ‘They argue about what it is and what the structure is.’ Police are worried about a growing number of young men choosing to invade underground rail networks to take photos. Documentary maker Adrian Ortega, 23, spoke with four men involved in 'urban exploring' and got four different answers about what it is. Two men who featured in the 30-minute film, Modus Operandi: A Look Into Melbourne's Urban Exploring Scene, are secretive urban explorers who go by pseudonyms and wear bandannas over their face to avoid being identified. They are the type of 'explorers' police are most concerned about. Photographer RDXV runs a Tumblr and Instagram account where he shares his photos from the underground rail tunnel networks of Melbourne and Sydney. ‘The most exhilarating part is when you're down in the loop and police are coming maybe, and you don't know what it is, something could happen at any time - that's the best part about it,’ he revealed in the documentary. In the 30-minute film, Modus Operandi: A Look Into Melbourne's Urban Exploring Scene, the urban explorers share their photos and memories. Photographer RDXV (left) and urban explorer Olympic Donuts (right) both appear in the documentary with face coverings. ‘When I was up in Sydney doing the loop my mate slipped over when a train was coming and could have been crushed by the train but he got up,’ RDXV added. For him urban exploring is about taking photos but also ‘having fun’. But that doesn’t mean he doesn’t get scared. ‘This one time I was with this girl on a rooftop and she climbed out on this cleaning rope support thing and was hanging there and I took a photo of her there. That was the stupidest thing I've seen anyone do, that was 20-storeys up,’ he said. Fellow ‘tunnel rat’ nicknamed Olympic Donuts, 16, said he got into urban exploring after 'walking around suburbs and getting into sticky situations’. ‘My friends started getting into rooftops and tunnels and they got me into it basically,’ Olympic Donuts said. Some of the men are secretive urban explorers who go by pseudonyms and wear bandannas over their face to avoid being identified. Some of the men have been caught by police before while others are yet to be found. Victoria Police claim they caught 1000 trespassers on Melbourne's City Loop in seven months last year. The urban explorers use whatever they can get their hands on to gain access to places they deem interesting. 'There are lots of tools to get in, coat hangers, lock picks,’ he explained. ‘You have to be stealthy, quick fast in and out, know what you are going in for.’ 'If you get caught you're going to get big fines or imprisonment it's scary but it also amps you up,’ Olympic Donuts reasoned. The other two men filmmaker Adrian spoke with are not involved with breaking into train tunnels and prefer to spend their time on construction sites and high-rise buildings. Bryce Wilson, nicknamed the Australian Spider Man, climbs structures 300 meters above Melbourne’s CBD to document people at their best and worst. ‘It's documenting and showing people things they might never have seen before and giving them a new appreciation of the city,’ Bryce said in the film. He doesn’t believe graffiti should be part of the unusual hobby. Although he does confess ‘the most exhilarating part is definitely avoiding police or security guards’. In this photo two men stand on the tracks inside Melbourne's city loop underground train tunnel system. RDXV believes 'the most exhilariting part is when you're down in the loop and police are coming' 'If you don't have any photos or video of it then you don't have no proof,' one urban explorer says in the documentary. 'It wouldn't be urban exploring without social media... if you're doing something you need to show it off,' Olympic Donuts said. ‘Bryce does it from a photojournalist view point… he’s totally against breaking into the City Link tunnels,’ Adrian explained. When Adrian asked Bryce if he does urban exploring for the adrenaline rush, Bryce denied that was the main kick he got out of it. Then there’s Lucas Bruce, who famously fell about 15 metres through the glass ceiling of Melbourne's Block Arcade in 2013 while ‘exploring’. Lucas reportedly lost his footing while climbing onto a ledge and fell nine metres through the glass roof and another six metres before smashing on the ground. ‘I tried to find the most legal way into buildings... making sure the buildings are unlocked,’ Lucas told Adrian. Bryce Wilson (left), nicknamed the Australian Spider Man, climbs structures 300 meters above Melbourne’s CBD to document people at their best and worst. Lucas Bruce (right) famously fell about 15 metres through the glass ceiling of Melbourne's Block Arcade in 2013 while 'exploring'. Neither are involved with graffiti or breaking into the city train loops. Bryce Wilson gets a kick out of taking photos from perspectives ordinary people don't get to see. The urban explorers all share a head for heights but documentary maker Adrian Ortega is scared of them. Construction sites and derelict buildings are some of the favourite places for urban explorers to take photos. Adrian was inspired to make the film after he previously documented Melbourne’s alternative nightclubbing scene and is not involved in urban exploring himself. ‘I followed a couple of the guys on Instagram… I’m afraid of heights but I got romantic and imagined what it would be like to do it.’ Asked if the men fear the police, Adrian said: ‘Two of them have been caught but the two other guys, I don't know their names, they are wearing masks and cover their identities – they haven’t been caught yet.’ Victoria police said in a statement: 'We take public safety extremely seriously and work closely with Metro Trains and other service providers to investigate and prosecute behaviour wherever and whenever it is detected. 'Depending on the incident, those detected may face serious criminal charges such as conduct endangering life and trespass.' Police warned trespassers on the rail networks can also be charged with: 'Interfering with automatic doors of a vehicle without reasonable excuse which carries a penalty of up to $283. 'Entering onto part of a rail vehicle not designed for carriage while vehicle is in motion which carries a penalty of up to $362. Travel on part of vehicle not meant for travel carries a penalty of up to $362. And moving, operation, interfering or tampering with equipment or vehicle without permission of an authorised adult carries a penalty of up to $362.' Adrian Ortega's documentary Modus Operandi: A Look Into Melbourne's Urban Exploring Scene premiers on Wednesday. To find out more visit his Facebook page or website. For more of RDXV's urban explorer photos, visit his Tumblr page

Summary: Police are concerned about people breaking into underground rail networks and disrupting services. Victoria Police claim they caught 1000 trespassers on Melbourne's City Loop in seven months last year. 'Those detected may face serious criminal charges such as conduct endangering life and trespass,' police warned. Documentary maker Adrian Ortega, 23, spoke with four men involved in 'urban exploring' Two of the men chose to remain nameless because they are involved in graffiti and photographing the rail networks. Another two men who spoke are against graffiti and prefer to take photos from tall buildings for their art.

cnn_dailymail

en

Write a title and summarize: Buruli ulcer (BU) caused by Mycobacterium ulcerans is a devastating skin disease, occurring mainly in remote West African communities with poor access to health care. Early case detection and subsequent antibiotic treatment are essential to counteract the progression of the characteristic chronic ulcerative lesions. Since the accuracy of clinical BU diagnosis is limited, laboratory reconfirmation is crucial. However, currently available diagnostic techniques with sufficient sensitivity and specificity require infrastructure and resources only accessible at a few reference centres in the African endemic countries. Hence, the development of a simple, rapid, sensitive and specific point-of-care diagnostic tool is one of the major research priorities for BU. In this study, we have identified a previously unknown M. ulcerans protein, MUL_3720, as a promising target for antigen capture-based detection assays. We show that MUL_3720 is highly expressed by M. ulcerans and has no orthologs in other prevalent pathogenic mycobacteria. We generated a panel of anti-MUL_3720 antibodies and used them to confirm a cell wall location for MUL_3720. These antibodies could also specifically detect M. ulcerans in infected human tissue samples as well as in lysates of infected mouse footpads. A bacterial 2-hybrid screen suggested a potential role for MUL_3720 in cell wall biosynthesis pathways. Finally, we demonstrate that a combination of MUL_3720 specific antibody reagents in a sandwich-ELISA format has sufficient sensitivity to make them suitable for the development of antigen capture-based diagnostic tests for BU. Buruli ulcer (BU) is a neglected mycobacterial skin disease, reported from tropical and subtropical countries world-wide with highest incidence rates in Western Africa [1]. Populations in rural areas with limited access to health facilities are most affected and often seek medical advice at late disease stages [2]. Advances in the clinical management of BU have shifted options for treatment from surgical resection to combination antibiotic therapy [1]. While PCR analysis targeting the insertion sequence IS2404 has evolved into the gold standard for laboratory diagnosis of BU, this test is only available at a few reference centres. Therefore, the diagnosis of BU is currently often based on clinical findings and antibiotic therapy is started before laboratory diagnostic results can be obtained. BU has a wide range of clinical manifestations including non-ulcerative forms such as subcutaneous nodules or papules, plaques and oedema, which may progress to chronic ulcerative lesions. Due to this diversity of disease presentations the accuracy of clinical diagnosis is limited [1,3–5] and thus a significant proportion of patients reporting with skin lesions may not receive adequate treatment. This includes cases of cutaneous tuberculosis which may be misdiagnosed as BU and thus receive the recommended eight week course of Streptomycin/Rifampicin combination chemotherapy for BU [5], which is much too short for the treatment of tuberculosis. As for IS2404 PCR, two of the other three currently applied methods for laboratory reconfirmation of BU—histopathology and cultivation of the extremely slow-growing mycobacteria—equally require expensive equipment and expertise [4,6–8] not accessible at peripheral health facilities. The only available point-of-care diagnostic test, direct-smear examination by microscopy for the detection of acid fast bacilli (AFB), has limited sensitivity and specificity [6]. Hence, one of the major research priorities for BU is the development of a fast, low-tech, sensitive and specific point-of-care diagnostic test, which can be directly implemented at peripheral health centres. The development of a specific point-of-care diagnostic test for the detection of M. ulcerans is complicated by the broad antigenic cross-reactivity among the various mycobacterial species. Serological approaches targeting the few M. ulcerans-specific antigens identified, turned out to be not suitable for differentiation between BU patients and exposed control individuals, as both groups may or may not exhibit serum IgG titers against these antigens [9–11]. In recent years, point-of-care tests in the form of antigen capture assays have successfully been developed for tropical infectious diseases [12]. Extensive studies focussing on rapid diagnostic tests for malaria [13–17] have paved the way for the development of antigen capture based assays for other diseases such as dengue fever [18,19], hepatitis C [20,21] or visceral leishmaniasis [22] to name but a few. In the present work we aimed at the identification of targets for the development of an antigen capture test for the diagnosis of M. ulcerans infection by using a proteomics approach. Ethical clearance for the analysis of clinical specimens was obtained from the Cameroon National Ethics Committee (N°172/CNE/SE/201) and the Ethics Committee of Basel (EKBB, reference no. 53/11). Immunization of mice for the generation of monoclonal antibodies was performed in strict accordance with the rules and regulations for the protection of animal rights (“Tierschutzverordnung”) of the Swiss “Bundesamt für Veterinärwesen”. All animal infection experiments performed were approved by the animal welfare committee of the Canton of Vaud (authorization number 2261) and were conducted in compliance with the Swiss animal protection law under BSL-3 conditions. In this study we analyzed M. ulcerans isolates from Ghana (NM20/02), Côte d’Ivoire (ITM 940511), Togo (ITM 970680), China (ITM 98912), Japan (ITM 8756) and Australia (JS5147) as well as additional mycobacterial strains including M. abscessus (ATCC 19977), M. avium (MAC101), M. chelonae (DSM 43804), M. fortuitum (ATCC 49403), M. gordonae (Pasteur 14021. 001), M. haemophilum (ATCC 29548), M. intracellulare (clinical isolate), M. kansasii (NCTC 10268), M. lentiflavum (clinical isolate), M. malmoense (NCTC 11298), M. marinum (ATCC 927), M. scrofulaceum (Pasteur 14022. 0031), M. simiae (clinical isolate) M. smegmatis (Pasteur 14133. 0001), M. terrae (clinical isolate), M. xenopi, M. bovis (ATCC 35734) and M. tuberculosis (Pasteur 14001. 0001). M. ulcerans strains were grown in BacT/Alert culture bottles supplemented with enrichment medium according to the manufacturer’s protocol (bioMérieux). For the preparation of M. ulcerans protein lysates, bacteria (5 ml of culture, OD600~1) were washed in PBS, heat-inactivated at 95°C for 35 min, centrifuged at 10′000 × g for 10 min and resuspended in 400 μl lysis buffer (PBS containing 5% SDS, 1 mM phenylmethylsulfonyl fluoride (PMSF) and a protease inhibitor cocktail (complete mini, Roche) ). The mix was transferred to lysing tubes (Precellys) and homogenized using a mechanical bead beater device (Precellys 24, Bertin Technologies) twice at 6′800 rpm for 30 s. Beads and non-lysed cells were removed by centrifugation at 10′000 × g for 10 min. The preparation of lysates of other mycobacterial species was described previously [9]. 90 μg of trichloroacetic acid (TCA) precipitated M. ulcerans (NM20/02) protein lysate was resuspended in rehydration buffer (8 M urea, 2% 3-[ (3-Cholamidopropyl) -dimethylammonio]-1-propanesulfonate (CHAPS), 0. 5% (v/v) ZOOM Carrier Ampholytes (Invitrogen), 0. 002% bromophenol blue and 0. 4% dithioerythritol (DTE) ). The mix was incubated with a 3–10 pH gradient strip (ZOOM Strip; Invitrogen) over night (ON) at room temperature (RT). First-dimension isoelectric focusing (IEF) was performed on a ZOOM IPG runner (Invitrogen) using a step voltage protocol (175 V for 15 min, 175–2000 V for 45 min, 2000 V for 2 h). After IEF, the strips were incubated for 15 min with equilibration buffer (6 M urea, 50 mM Tris pH 8. 8,30% glycerol, 2% SDS, 30 mM DTE) followed by a 15 min incubation period with alkylating solution (6 M urea, 50 mM Tris (pH 8. 8), 30% glycerol, 2% SDS, 0. 23 M iodacetamide). Second-dimension gel electrophoresis was performed at 200 V for 35 min using a 10% NuPAGE Novex Bis-Tris ZOOM Gel (Invitrogen). The gel was stained with Coomassie blue (Invitrogen). All Coomassie stained protein spots were selected for mass spectrometry analysis. Spots were excised from the 2D gel, placed in a low-binding microcentrifuge tube and destained in 0. 1 M ammonium bicarbonate / 30% acetonitrile at 30°C. Gel spots were dried in a SpeedVac concentrator and digested with 4 μl of 10 μg/ml trypsin (trypsin porcine, Roche Applied Science) ON at 37°C. Peptides were extracted from gel pieces with 4 μl of 0. 3% trifluoroacetic adic (TFA) / 50% acetonitrile. The samples were desalted and concentrated using ZipTipC18 tips (Millipore). Eluted peptides were loaded onto a MALDI target. MS analysis was performed using a MALDI-TOF mass spectrometer (Bruker ultraflex III TOF/TOF, Bruker Daltonics Inc.) in the reflector mode. 1 μl of tryptic digest and 1 μl of matrix (5 mg/ml α-cyano-4-hydroxycinnamic acid, 50% acetonitrile, 0. 1% TFA) were spotted onto a MALDI target (MTP AnchorChip 600/384, Bruker Daltonics) and allowed to co-crystallize at room temperature. Data were processed using FlexAnalysis software (Bruker Daltonics flexAnalysis 2. 4). Spectra were smoothed (Sawitzgy Golay algorithm, 0. 2 m/z width, 1 cycle), baseline subtracted (median algorithm, 0. 8 flatness) and calibrated using trypsin autocleavage or internal standard peptide mass peaks. A monoisotopic peak list was generated from the spectrum using SNAP algorithm and analyzed with BioTools (Bruker Daltonics BioTools 3. 0). Peptide mass fingerprinting searches were performed using the Aldente search engine on the Expasy server. The full length MUL_3720 (aa 1–207) and a truncated version of this protein lacking the lectin domain (aa 115–207) were recombinantly expressed in Escherichia coli BL21 Star (DE3, Invitrogen) as N-terminal hexahistidin-tagged fusion proteins. Briefly, PCR was performed on a pUC57 vector containing the DNA sequence of MUL_3720 generated by gene synthesis (Genscript), including NdeI and NotI restriction sites. The amplified sequences were inserted into a TOPO-TA cloning vector using the TOPO Cloning Kit and introduced into E. coli (Top 10, Invitrogen). The vector was digested with NdeI and NotI (New England Biolabs) and the sequence was ligated into a pET28a expression vector using the Rapid DNA Ligation Kit (Roche). E. coli BL21 Star (DE3, Invitrogen) were grown in Luria-Bertani (LB) medium until an OD600 of ~0. 5. Protein expression was induced by addition of isopropyl thiogalactoside (IPTG) to a final concentration of 1 mM and subsequent incubation for 3 h at 37°C. Bacteria were lysed by sonication and His-tagged proteins were purified by nickel-nitrilotriacetic acid (Ni-NTA) chromatography. MUL_3720 was amplified from genomic M. ulcerans DNA and cloned into TOPO vector using NdeI and ScaI restriction sites. Electrocompetent E. coli TOP10 cells (Invitrogen) were transformed with TOPO: : MUL_3720 vectors and spread onto LB-Ampicillin (50 μg/ml) agar. Plasmid DNA of the mutant colonies was prepared and inserts with correct size and sequence were excised from TOPO by NdeI/ScaI and ligated into the mycobacterial vector pSD5. Chemically competent E. coli TOP10 were transformed with pSD5: : MUL_3720 and grown on LB-Kanamycin (50 μg/ml) plates. Plasmid DNA was prepared and the presence of the insert was confirmed. M. ulcerans strain NM20/02 was grown in BacT bottles (bioMérieux) containing enrichment medium (bioMérieux). Bacteria were harvested and washed twice with 10% glycerol or distilled water. Competent M. ulcerans bacteria were electroporated (2. 5 kV, 1000 Ohm, 25 μF) with varying amounts (50–1000 ng) of DNA, transferred to MGIT-OADC medium (BD) and grown under non-selective conditions for 36 hours at 30°C. After the recovery phase, bacteria were spread on 7H10-Kanamycin (25 μg/ml) agar and incubated for several months at 30°C. Colonies were picked and regrown on selective agar and in BacT bottles (bioMérieux) in order to prepare lysates and stocks. For the preparation of mAbs, mice were immunized two times intraperitoneally with 40 μg of recombinant full length MUL_3720 (aa 1–207) emulsified in Immune Easy adjuvant (Qiagen). Two weeks after the second immunization serum antibody titres against MUL_3720 (aa 1–207) as well as against the truncated MUL_3720 (aa 115–207) were determined by ELISA. Based on these results one BALB/c mouse was selected to receive a final intraperitoneal injection of 40 μg of recombinant MUL_3720 (aa 1–207) without adjuvant. Three days after this last booster dose, hybridoma cell lines were generated as described previously [23]. Briefly, the spleen of the selected mouse was removed and the spleen cells were fused with mouse myeloma cells (PAI cells). After a few days, cell culture supernatants were tested for the presence of anti-MUL_3720 (aa 1–207) as well as anti-MUL_3720 (aa 115–207) antibodies. Positive cell lines were cloned by limiting dilution and expanded. MAbs were purified using HiTrap rProtein A column (Amersham Biosciences). Two individual fusion experiments resulted in 24 and 17 MUL_3720-ELISA positive B-cell hybridoma cell lines, respectively. Of these, a total of 5 B-cell hybridoma cell lines (JD3. 1, JD3. 2, JD3. 3, JD3. 4, JD3. 6, JD3. 7) were successfully cloned and expanded for antibody production. Rabbit polyclonal antibodies were generated and affinity purified by Eurogentech. New Zealand white Rabbits were injected intramuscularly with 20 μg recombinant MUL_3720 (aa 1–27) with Sigma Adjuvant System (SZ3398) or Imject Alum (SZ3403) on day 0,14,28 and 56. Total IgG was purified from antiserum collected on day 66 by protein A affinity chromatography. 96-well Immulon microtiter plates (Thermo Scientific) were coated with 1 μg recombinant MUL_3720 (aa 1–207) or MUL_3720 (aa 115–207) per well in 100 μl PBS and incubated ON at 4°C. Plates were washed three times with washing buffer (2. 5% Tween 20 in dH2O) and blocked with 5% non-fat dry milk in PBS containing 0. 1% Tween for 1 h at 37°C. After washing as described above, 100 μl of the primary antibody (mAbs or hybridoma supernatant) was added and incubated for 2 h at 37°C. Following an additional washing step, 100 μl of 1: 30′000 diluted goat anti-mouse IgG (γ-chain specific) antibodies coupled to alkaline phosphatase (SouthernBiotech) was added to each well and incubated for 1 h at 37°C. Plates were washed and 100 μl/well of phosphatase substrate solution (1 mg/ml p-Nitrophenyl phosphate in substrate buffer) was added and incubated for 1 h at 37°C. Absorbance at 405 nm was measured with a microplate reader (Tecan Sunrise). 2 μg of mycobacterial protein lysates per lane were separated on NuPAGE Novex 4–12% Bis-Tris ZOOM Gels (Invitrogen) using NuPAGE MES SDS Running Buffer (Invitrogen) under reducing conditions. After electrophoresis proteins were transferred onto nitrocellulose membranes using an iBlot gel transfer device (Invitrogen). Membranes were blocked with blocking buffer (5% non-fat dry milk in PBS) ON at 4°C. Membranes were then incubated in blocking buffer containing anti-MUL_3720 IgG (mouse mAbs JD3. 2, JD3. 4 or rabbit polyclonal IgG SZ3398) or mouse mAb DD3. 7 (specific for a conserved mycobacterial protein) serving as loading control for 1 h at RT. After washing, membranes were incubated with secondary goat anti-mouse IgG (γ-chain specific) (HRP, SouthernBiotech) or goat anti-rabbit IgG (Fc fragment specific) (HRP, Milan) for 45 min at RT. After washing, bands were visualized by chemiluminescence using the ECL Western Blotting substrate (Pierce). Immunohistochemical analysis was performed on tissue or punch biopsies from different IS2404 qPCR reconfirmed patients. Tissue or punch biopsies of BU patients were removed aseptically and immediately fixed in 10% neutral buffered formalin for 24 hours. Afterwards the tissue was embedded into paraffin, cut into 5 μm thin sections and transferred onto microscopy glass slides. Immunohistochemical staining of the sections was performed after deparaffinisation, rehydration and antigen retrieval with citrate-pretreatment according to standard protocols [24]. Inactivation of endogenous peroxidase as well as prevention of unspecific binding was achieved by incubation in PBS containing 0. 3% hydrogen peroxide and 1. 5% horse serum for 20 min. Primary anti-MUL_3720 IgG was diluted in PBS containing 0. 1% Tween-20 and added to the slides for 1 h at RT or ON at 4°C. After incubation with biotin-conjugated horse anti-mouse IgG, slides were stained using the Vector ABC and NovaRED system. Sections were counterstained with haematoxylin. JD3. 4 and JD3. 2 showed a comparable staining in intensity, specificity and sensitivity. JD3. 2 gave a slightly lower unspecific background staining of the surrounding tissue and was used for IHC analysis. IFA was carried out as described previously [25]. Briefly, a pellet of M. ulcerans bacteria (OD600~0. 6) was resuspended in 1. 5% low-melting agarose (BioWhittaker Lonza, Basel Switzerland) and transferred to cryomodules (Applied BioSystems). Agarose blocks were embedded into paraffin, cut in 3 μm sections and transferred onto microscopy glass slides (Thermo Scientific). Bacteria were stained with mAb JD3. 2 and Alexa fluor488 (Invitrogen) conjugated goat anti-mouse IgG and mounted in ProLong Gold anti-fade reagent containing 4′, 6-Diamidino-2-phenylindole (DAPI; Invitrogen). The system described for investigating protein interactions by the functional reconstitution of a murine dehydrofolate reductase domain in M. tuberculosis [26] was modified here for use in M. ulcerans. N-terminal and C-terminal fusions of the bait domains to full length MUL_3720 were constructed using pUAB400 or pUAB200. Cloning was facilitated by the MfeI and ClaI restriction sites in the pUAB multiple cloning sites. All cloning was performed using E. coli DH10B and confirmed by Sanger sequencing. Plasmids were extracted from E. coli using mini-prep columns (Qiagen) and plasmid DNA was used to transform M. smegmatis MC2155 by electroporation as previously described [27]. M. ulcerans Agy99 genomic DNA libraries were prepared by partial AciI digestion. Digested DNA between 500 bp and 3 Kbp was purified using a gel purification kit (Qiagen), ligated into ClaI digested pUAB300 and used to transform E. coli DH10B. A number of colonies were randomly selected for PCR using primers F102 (5′-agaaccaccacgaggagctcat-3′) and R102 (5′-tgatgcctggcagtcgatcgta-3′) that flank the multiple cloning site on the vector to check for insertions containing inserts within the desired size range [26]. Approximately 2 × 105 clones were subsequently collected and cultured in LB ON. Plasmid DNA maxi-preps were performed on ON cultures according to the manufacturer’s instructions (Sigma-Aldrich). Bacteria co-transformed with plasmids containing interacting, complementary mDHFR fragments were selected on 7H11 kanamycin (25 μg/ml) and hygromycin (50 μg/ml) plates. Colonies were patched onto 7H11 kanamycin-hygromycin-trimethoprim plates and colonies resistant to trimethoprim were selected for PCR. Using primers F102 and R102, PCR products were sequenced. The sequences were used to perform BLAST against the M. ulcerans Agy99 genome. Inserts containing open reading frames in the incorrect orientation were discarded. Also removed were inserts that matched non-coding genomic DNA or the dehydrofolate reductase from M. ulcerans. M. ulcerans strain S1013 used for experimental infection of mice was isolated in 2010 from the ulcerative lesion of a Cameroonian BU patient [28]. Bacteria were cultivated in Bac/T medium for 6 weeks, recovered by centrifugation and a stock suspension in sterile PBS of 125 mg/ml wet weight was prepared. 30 μl of a 1: 1000 dilution of the stock solution was injected subcutaneously into the left hind foot pad of 14 week old female BALB/c mice. On day 87 after infection, mice were euthanized and foot pads were aseptically removed. Foot pads were dipped into 70% ethanol, dried under the laminar flow, cut into four pieces with a scalpel and transferred to reinforced hard tissue grinding tubes (MK28-R, Precellys) containing 750 μl of Bac/T medium (bioMérieux). Tissue homogenization was performed with a Precellys 24-dual tissue homogenizer (3 × 20 s at 5000 rpm with 30 s break). After transferring the supernatant to a fresh tube, the residual tissue remains were homogenized a second time in 750 μl of Bac/T medium. Tissue lysates were pooled and stored at -80°C until further use. 500 μl of thawed tissue lysate was transferred into tough microorganism lysis tubes (VK05–2ml, Precellys), inactivated for 1 h at 85°C and centrifuged at 17′000 × g for 5 min. The pellet was resuspended in 250 μl PBS containing protease inhibitors (Roche, EDTA—free) and cells were disrupted with Precellys 24-dual tissue homogenizer (2 × 30s at 6800rpm with 1 min break in between). Lysates were cleared by centrifugation and tested by ELISA. Nunc-Immuno Maxisorp 96-well plates (Thermo Scientific) were coated with 10 μg/ml JD3. 4 mAb (50 μl per well) in PBS and incubated ON at 4°C. Plates were washed three times with washing buffer (2. 5% Tween 20 in dH2O) prior to incubation with blocking buffer (5% non-fat dry milk in PBS) for 2 h at RT. After washing as described above, 50 μl of different dilutions of the purified recombinant full length MUL_3720, M. ulcerans lysate (NM20/02), or lysates from M. ulcerans infected tissue samples in PBS were added and incubated for 2 h at RT. Following an additional washing step, 50 μl anti-MUL_3720 rabbit IgG (5 μg/ml) in blocking buffer with detergent (0. 5% non-fat dry milk in PBS containing 0. 05% Tween 20) was added and incubated for 2 h at RT. After washing as described above, 50 μl goat anti-rabbit IgG coupled to horseradish-peroxidase (Milan) diluted 1: 10′000 in blocking buffer with detergent was added and incubated for 1 h at RT. Plates were washed and TMB peroxidase substrate solution was added. After 10 min the reaction was stopped with 2 M sulfuric acid and absorbance was measured at 450 nm with a microplate reader (Tecan Sunrise). For identification of suitable proteins that could be used as targets in diagnostic test formats, an M. ulcerans whole protein lysate was analysed by 2D gel electrophoresis (S1 Fig.). In total, 384 protein spots were detected, processed and subsequently subjected to MALDI-TOF-MS. Among the 384 spots, 118 peptide fragments were identified and attributed to 36 different genes. In order to select for proteins without orthologs in M. tuberculosis, M. bovis or M. leprae, a BLAST search against the Uniprot database was performed for all 36 proteins, resulting in the identification of three potential targets (MUL_3720, MUL_0343 and MUL_4023) suitable for a selective antigen capture assay. However, MUL_0343 and MUL_4023 presented very weak protein spots in the 2D gel, while MUL_3720 showed a high expression level and was therefore selected for further analysis. The 624 bp MUL_3720 gene encodes a protein of 207 amino acids, with a molecular mass of 22 kDa. MUL_3720 is predicted to possess an N-terminal bulb-type mannose-specific lectin domain and a C-terminal peptidoglycan-binding Lysin Motif (LysM) linked by a proline-rich sequence (Fig. 1). Database comparisons revealed the presence of orthologs with a similar domain organisation in M. abscessus (MAB_2373), M. avium, M. colombiense, M. fortuitum, M. kansasii (MKAN_05370), M. marinum (MMAR_3773), M. smegmatis (MSMEG_3662) and M. xenopi. The M. marinum ortholog displayed a sequence identity of 99% (S2 Fig.). For the generation of antibodies against MUL_3720, required for the detection of the protein in diagnostic assays, we immunized mice and rabbits with the full length protein, recombinantly expressed as a His-tagged fusion protein (predicted molecular mass 24 kDa) in E. coli BL21. Hybridoma cell lines producing antibodies against different epitopes of the protein were identified by analyzing their reactivity against MUL_3720 (aa 1–207) as well as the truncated version of MUL_3720 (aa 115–207), lacking the lectin domain and consisting only of the LysM motif and the proline-rich sequence. Five mAbs (JD3. 2, JD3. 3, JD3. 4, JD3. 6 and JD3. 7), all of them mouse IgG1 (κ) isotype, were generated, purified and further characterized. All the mAbs recognized recombinant full length MUL_3720 (aa 1–207) in ELISA. While all antibodies except for JD3. 6 recognized recombinant full length MUL_3720 (aa 1–207) in Western Blot analysis (Fig. 2A), only JD3. 2 and JD3. 4 also reacted with recombinant truncated MUL_3720 (aa 115–207) (Fig. 2B) and the endogenous protein in M. ulcerans lysates (Fig. 2C) (S1 Table). The ability of mouse mAbs JD3. 2 and JD3. 4 to detect endogenous MUL_3720 in lysates of M. ulcerans strains from different geographical regions was examined by Western Blot analysis. While the protein was recognized by JD3. 2 and JD3. 4 in all M. ulcerans strains, isolates belonging to the classical M. ulcerans lineage (Ghana, Côte d’Ivoire, Togo and Australia) showed higher expression levels as compared to isolates belonging to the ancestral lineage (China and Japan) (Fig. 3). Interspecies cross-reactivity of MUL_3720 was determined by Western Blot analysis with lysates of a range of different mycobacterial species (Fig. 4). In accordance with the BLAST search for MUL_3720 orthologs (S2 Fig.), rabbit polyclonal anti-MUL_3720 IgG reacted with proteins in lysates of M. fortuitum, M. marinum, M. smegmatis and M. xenopi. In agreement with a shorter linker between the lectin and the LysM domains (S2 Fig.), the M. xenopi ortholog was detected at a lower molecular weight. The predicted orthologous proteins in M. abscessus, M. avium and M. kansasii were not recognized by the rabbit polyclonal IgG. Furthermore, a protein band was observed in lysates of M. gordonae, M. malmoense and M. terrae for which no sequence information is available (Fig. 4A). The detected proteins in M. malmoense and M. terrae were slightly smaller than MUL_3720. Analysis with JD3. 4 led to a similar staining pattern among the mycobacterial lysates, except for the protein expressed by M. gordonae, which was not recognized (Fig. 4B). JD3. 2 only reacted with protein in lysates of M. malmoense and M. marinum, suggesting that the two mAbs JD3. 2 and JD3. 4 recognize different epitopes of MUL_3720 (Fig. 4C). In order to confirm the expression of MUL_3720 by M. ulcerans and the ability of the anti-MUL_3720 mAb JD3. 2 to detect the protein in vivo, we performed immunohistochemistry and immunofluorescence stainings. M. ulcerans could be detected in punch biopsies of human BU patients with the mAbs JD3. 2 (Fig. 5) and JD3. 4. ZN staining (Fig. 5 A, C, E) and JD3. 2 staining (Fig. 5 B, D, F) of serial sections showed identical localization at the same tissue region. AFBs detected by ZN staining in tissue sections revealed a homogeneous staining pattern, whereas immuno-staining with mAb JD3. 2 exhibited a heterogeneous staining pattern of the bacteria with intensively stained poles (Fig. 5), indicating a higher expression of the protein in these areas in the natural environment of the bacteria. In contrast, IFA of in vitro cultivated bacteria showed a more homogeneous distribution of the protein on the bacterial cell surface. This localization was confirmed in a MUL_3720 overexpressing M. ulcerans strain (Fig. 6). As a first step to begin to understand the role of MUL_3720 we used a bait and prey approach to identify other M. ulcerans proteins that interacted with this protein. We employed the mycobacterium-specific protein fragment complementation (M-PFC) system. An M-PFC bait clone using a N-terminal fusion of MUL_3720 and co-transformation with a random library of M. ulcerans genomic DNA fragments in pUAB300 (prey) resulted in approximately 150 trimethoprim resistant colonies. Subsequent clones were patched and screened using primers F102 and R102 to determine the identity of the DNA sequence present in pUAB300 (Table 1). Multiple independent clones were identified for sequences encoding DesA1 (MUL_0445) and a PE-PGRS protein (MUL_0572), together with 13 other single-hit CDS, including an interaction with MUL_3720 itself. Many of the putative interacting proteins had a predicted cell wall location or role in cell wall biosynthesis, in line with the localization data for MUL_3720 revealed by mAb staining (Table 1 and Fig. 6). No interacting proteins were identified using the C-terminal MUL_3720 bait fusion, consistent with the predicted cell wall location for this domain of MUL_3720. The M-PFC detects cytoplasmic protein-protein interactions only [26]. We analyzed different combinations of the generated mAbs and polyclonal IgG as MUL_3720 capturing and detecting antibodies in an antigen capture sandwich ELISA. The application of mAb JD3. 4 as capturing and polyclonal rabbit IgG as detecting reagent enabled a highly sensitive detection of recombinant MUL_3720 (Fig. 7A) and the endogenous protein present in lysates of in vitro cultivated M. ulcerans (Fig. 7B). In order to test if the antigen capture ELISA is able to detect MUL_3720 expressed by bacteria in infected tissue samples, we analyzed lysates of M. ulcerans infected mouse foot pads. MUL_3720 could be detected in lysates of all five infected tissue samples analyzed, while only background readouts were obtained for lysates of uninfected foot pads (Fig. 8). Attempts to develop a diagnostic tool based on serological approaches have been equivocal [9–11], so we decided to focus on direct detection of M. ulcerans antigens in BU patient specimens. In the present study, we identified the MUL_3720 protein as a promising target in antigen capture-based diagnostic tests for M. ulcerans. Based on 2D gel electrophoretic analyses, MUL_3720 is one of the most highly expressed proteins in vitro. The high expression of MUL_3720 is considered an advantage with respect to developing a sensitive antigen detection test for the diagnosis of BU. While the biological role of MUL_3720 is not known, clues to its function are suggested by its two-domain structure—a conserved bulb-type mannose-binding lectin domain and a Lysin Motif (LysM) domain—predicted to be involved in alpha-D-mannose recognition and in binding to peptidoglycan, respectively. Some bacterial species retain certain proteins attached to peptidoglycan by their LysM domains [29]. We used a mycobacteria-specific two-hybrid system to search for M. ulcerans proteins interacting with MUL_3720 and we had hits to a range of proteins known or predicted to be cell-wall associated or involved in cell wall synthesis (Table 1). Many of the interacting proteins—such as DesA1 (Table 1) —are involved in biosynthesis or modification of cell wall molecules. In other mycobacteria, the resulting double bonds from the DesA1-mediated catalysis of a desaturation reaction of saturated alkyl chains that arise during mycolic acid synthesis are required for subsequent position specific modifications such as epoxidation and cyclopropanation of this key cell wall metabolite [30,31]. MUL_3720 appears to be arranged in an operon structure with two adjacent putative cell wall-associated protein coding genes (MUL_3721, MUL_3722). Immunofluorescence stainings of M. ulcerans bacilli confirmed the cell wall localization of MUL_3720. With its cell wall location, the two-domain structure including a mannose-binding N-terminal cytoplasmic component and C-terminal peptidoglycan-binding component, its operon structure and a substantial list of potential interacting proteins, MUL_3720 may be an adaptor protein for multiple cell wall biosynthetic pathways. MUL_3720 might play a role in cell attachment and cell-cell interactions given its presence at the cell surface as revealed by immunofluorescence microscopy and immunohistochemical analyses. The cell-surface localization of MUL_3720 is an additional advantage with respect to developing a sensitive diagnostic test, since the protein is expected to be easily accessible and detectable in tissue specimens of BU lesions. Potential shedding of the protein from the cell surface may facilitate ready detection in body fluids, which will be examined in future experiments. Monoclonal and polyclonal antibodies against MUL_3720 were generated for the development of antigen capture assays. These antibodies recognized in vitro grown M. ulcerans bacilli as well as bacteria in biopsies of human BU patients, proving the expression of MUL_3720 in BU lesions. Since these antibodies did not react with orthologs of MUL_3720 in other pathogenic mycobacterial species prevalent in the BU endemic regions, prospects for the development of a test with the desired specificity, excluding in particular cutaneous tuberculosis [5], are good. The monoclonal antibodies used for the antigen capture test bind to an epitope on the proline/rich linker and/or the LysM domain. Since the LysM domain is a widespread protein module present in more than 4000 proteins of both prokaryotes and eukaryotes [29], the potential cross-reactivity of the anti-MUL_3720 antibodies with those proteins remains to be analyzed. Importantly, this capture assay specifically detected MUL_3720 protein in tissue lysates of M. ulcerans infected mouse footpads. Furthermore, initial results revealed that MUL_3720 could be detected in swab samples from human BU lesions with a high bacterial burden (manuscript in preparation). Ongoing optimization of the applied reagents as well as the assay format is aiming at the development of a simple test format appropriate for low-resource laboratory settings with suitable test sensitivity. Antibiotic treatment of BU in its early stages leads in most of the cases to complete healing of the lesions with little or no trauma, whereas treatment at later stages often requires adjunct surgical treatment and is associated with prolonged hospitalization and long-term sequelae. The development of a simple and rapid diagnostic test, whose key elements are provided in the work presented here, will be of immediate benefit to BU patients in rural endemic communities. Clinical findings could directly be reconfirmed by this point-of-care test helping to avoid a false diagnosis and to facilitate a prompt onset of adequate treatment.

Title: Identification of the Mycobacterium ulcerans Protein MUL_3720 as a Promising Target for the Development of a Diagnostic Test for Buruli Ulcer Summary: According to the recommendations of the World Health Organization, the clinical diagnosis of BU should be reconfirmed by at least two laboratory techniques. However, out of the four currently available tests, three (PCR, histopathology and cultivation of M. ulcerans) can only be performed at centralized reference laboratories; the fourth (microscopic detection of acid fast bacilli) lacks the required sensitivity and specificity. Therefore, a simple tool for early diagnosis of the disease, which can be implemented in rural health care facilities of the endemic countries, is of urgent need. In this study we aimed at the identification of M. ulcerans proteins as potential targets for the development of a simple and rapid diagnostic antigen detection assay. Among 36 proteins, MUL_3720 best met the predefined criteria of being highly expressed by M. ulcerans and not having orthologs in other pathogenic mycobacterial species prevalent in the endemic regions. Here we generated monoclonal and polyclonal antibodies against this protein and carried out pilot studies for the development of an antigen capture-based diagnostic test.

lay_plos

en

Summarize: By. Lucy Crossley. A primary school headteacher has apologised after parents complained that a project on the discovery of a giant 'alien egg' in the playground left children in tears and too afraid to go to school. The project at Holy Trinity Primary School in Halstead, Essex, revolved around a 3ft-high egg found in undergrowth on school grounds and was part of a problem solving exercise. Despite assurances that the 'amazing discovery' was '100 per cent safe', some of the school's 250 pupils were so frightened by the egg that headmaster Jon Smith had to issue an apology after parents reported their children were distressed, and even suffering from nightmares. Project: The headteacher of Holy Trinity Primary School in Halstead, Essex, has apologised after some pupils were frightened by a project involving the 'discovery' of a giant egg on school grounds. As part of the project, which took place last month, parents were sent letter reporting how the egg had been found, and that a scientist from the 'National Museum Of Strange Objects' was coming to investigate it. 'I am pleased to inform you that the. inspection has confirmed that the area and object are 100 per cent safe and pose. no threat whatsoever to the public,' Mr Smith wrote in his mock alert. 'What still isn’t clear however, is. the identity of the object.' He then said that the museum's Dr Violet Strangeways had asked that all children'remain vigilant and put all their. energies into research and investigation in order to help the museum. ascertain the origins of this amazing discovery'. Worry: Some parents at the school reported that their children had been upset by the 'discovery' of the egg. Apology: Headteacher Jon Smith sent out this apology to parents whose children had been worried by the egg stunt. As part of the project the children had to question where the egg might have come from, drawing pictures and writing essays on its provenance, with parents kept up to date on their project. However, a post on the school's Facebook page and Twitter account soon began attracting comments from some parents who reported that their children had been frightened. One parent wrote: '@HTPSHalstead tell me I'm not the only parent whose child hasn't eaten lunch & has been in tears about the "discovery"? Prank too far?' 'It has come to our attention that whilst. the vast majority of the children at school were excited and engaged by. this morning’s "discovery" there are some who have been worried by it' Jon Smith, Headteacher. Another added: '@HTPSHalstead glad you. said that my son does not want to go to school anymore and has been in. tears since he come home #nothappy.' Although. the majority of parents writing on the school's Facebook page said. their children had been excited by the project, with the pupils. speculating that the egg could be that of a dinosaur or dragon, others. said that some younger children had been scared by it. One commented: 'He is scared of his own shadow and has been to sleep earlier but woke up at 9.20pm after having a nightmare!' Others said their children had been afraid to go to bed, with one mother writing: 'Holy trinity..........i hope this doesn't go on all week!!!!! I'm gona have 2 tired children!!!!!!!' Within hours of the initial post, Mr Smith had issued another letter, apologising to parents if their children had been upset by the egg. 'It has come to our attention that whilst the vast majority of the children at school were excited and engaged by this morning’s "discovery" there are some who have been worried by it,' he said in a second letter issued on June 16. School grounds: The well-meaning project revolved around a 3ft-high egg found in undergrowth on school grounds and was part of a problem solving exercise. 'We assure you that this afternoon we held an extra whole school assembly where we made it absolutely clear to everyone that the egg was of no danger to anyone, that it was safe to keep in school, and that we would therefore be able to observe it further tomorrow. 'We apologise if that message did not come through sufficiently and, as planned, in the follow-up assembly in the morning we will place further emphasis on it being of absolutely no danger whatsoever. 'During the course of today the children have produced an enormous amount of high quality work, in a variety of forms, related to the egg and it was great to see them so engaged. Such "discoveries" are quite common in primary schools across the country and are a very successful way of promoting problem solving, teamwork, group discussion and PSHE-related topics. This activity is part of our "No Problem" theme this term.' Holy Trinity Primary School had not responded to MailOnline's requests for comment at the time of publication

Summary: Problem solving project centred on a 3ft-high egg found in school grounds. Children were told the egg at Holy Trinity Primary School in Halstead, Essex, was safe, and asked to help investigate the 'amazing discovery' Many parents said their children were enjoying the project. But some complained younger pupils were in tears or having nightmares. Headteacher Jon Smith later apologised if children were 'worried' by stunt.

cnn_dailymail

en

Summarize: Durbin: Reid, McConnell Working on Debt Deal File Photo Senate leaders are hashing out a plan this week to raise the debt ceiling and avert government default, Senate Majority Whip Dick Durbin (D-Ill.) said Sunday. “The good news is Majority Leader Harry Reid and Sen. Mitch McConnell are sitting down and working out an approach that we’re going to try to tackle this week,” Durbin, who participated in last week’s White House talks on raising the debt ceiling, told CBS’ “Face the Nation.” With 16 days left before the Treasury Department’s Aug. 2 deadline, talks are intensifying between Reid (D-Nev.) and Minority Leader McConnell (R-Ky.) on a plan based on a framework provided by the Republican leader. The Senate is poised to hold votes this week on a balanced budget amendment to the Constitution, and it’s likely to vote on the Republicans’ “cut, cap and balance” plan. Although neither measure is expected to pass, leaders hope the votes can clear the way for the ultimate package that would raise the debt ceiling before August and perhaps include a series of hard budget cuts being negotiated now. “We have to check the boxes. One of them is to engage in this debate,” Durbin said regarding why the Senate would vote on the balanced budget amendment even though it is destined to fail. But even as Reid and McConnell work on a package that could be palatable for both their caucuses, rank-and-file Senators continue to express reservations about McConnell’s “Plan B,” which would raise the debt limit in a series of three votes that put the burden on President Barack Obama and the Democrats but do not require hard cuts. Sen. Tom Coburn said Sunday he was “unlikely” to vote for any iteration of the McConnell plan. The Oklahoma Republican walked away in May from the Senate’s bipartisan “gang of six,” which had been working for months on a $4 trillion package of deficit reductions. Durbin is one of the remaining five members. “The McConnell plan is more of Washington not taking responsibility. It’s a great political plan — it takes the pressure off all the politicians but allows us to pass a debt limit [extension] without making the hard choices that this country has to make,” Coburn said on “Face the Nation.” “I haven’t firmly decided, but I’m unlikely to support it at this time,” the Oklahoma Republican added. Coburn is still advocating for savings that he called “difficult, but not super hard,” such as cutting $1 trillion from the Pentagon budget over the next 10 years. PRES. OBAMA: I've put things on the table that are important to me and to Democrats, and I expect Republican leaders to do the same. MR. GREGORY: As deliberations continue, there were no face-to-face talks this weekend, and House Republicans are expected to vote Tuesday on a series of measures to cut spending and balance the budget, although they don't appear to have the votes necessary for those measures to become law. Joining me now, the president's top budget adviser Jack Lew. Welcome to MEET THE PRESS. MR. JACK LEW: Good to be here, David. MR. GREGORY: Good to have you here. So what is the latest? Have there been substantive talks over the course of the weekend? MR. LEW: Well, the latest is that, after the meeting on Thursday, there've been a lot of conversations within each of the party caucuses, within the House and the Senate, phone calls and conversations back and forth. That's how the president left it on Thursday, that we'd kind of put all of the items on the table. It was now a question of Congress figuring out what it could do. So I think that will continue over the next day or so. MR. GREGORY: But you don't have, as we sit here now, a, a better sense of what Congress is willing and able to do. MR. LEW: I think that what is encouraging is that the leaders in Congress seem to all agree that we can't push to a default, that we need to have a path that makes sure that the United States can keep its obligations and pay its bills in August. So I think that there are many conversations going on in order to make sure that that doesn't happen. MR. GREGORY: Well, let me be clear. Is the president's position that he would accept no more than $1 trillion in cuts if there's no tax increases? Is that the number that he's sort of dealing with in his head? MR. LEW: I, I think, I think it's a little less mechanical than that. The president made clear he wants the largest deal possible. He wants to do the most we can to reduce the deficit. That would be the right thing to do for the American people. He made it clear he's willing to go into areas that he's not in the past been comfortable going into, and others will have to do that as well. MR. GREGORY: But he used that figure. MR. LEW: But he also said that if we can't get the most done, then in addition to extending the debt we should do as much as we can. There are a number of ways to get there. We aspire to, we aspire to more than -- yeah. MR. GREGORY: But -- sure, but, but my question is the number. A -- if you get beyond $1 trillion and he still can't get any tax increases, the president said, "I'm not willing to sign something that, that cuts more spending without having tax increases." MR. LEW: Well, what, what, what the president said was to do major structural changes on the spending side, there would have to be tax increases. And I think that one can see a path to getting well over $1 trillion on things that we should be able to agree to. MR. GREGORY: Hm. So if that's something of a fallback plan, because the president wanted something on the order of $4 trillion over 10 years of spending cuts, Erskine Bowles told my colleague Chuck Todd on his program, "The Daily Rundown," that that might fall well short of what's necessary here. This is what he said. MR. ERSKINE BOWLES: The problem is, Chuck, that so many people are talking about doing it with just about $2 trillion of deficit reduction. That's not a solution. That's not going to fool our creditors. MR. GREGORY: In other words, the markets, our creditors are going to look at that and say that's not a fallback position, that's well short of what the United States should be doing. MR. LEW: I think if you look at what the markets are saying, they're saying two things. They're first saying the United States cannot default. And I think that is the bare minimum. And I say that again only because it has to be clear that there's some extreme views in some places that think that that's something we can do. We can't. The second is we need to get our fiscal house in order. I think that we've said for some time now, as have most, that we need to do on the order of $4 trillion of deficit reduction over the next 10, 12 years. We would like to get that done now. If that can't happen, if there's not a willingness to come together, the president has shown he's willing to make the kinds of, of moves that are necessary to get there. But if there's not a similar willingness, we should do as much as we can now. I think that the markets will understand moving far -- as far as we can. What will be hard to explain is doing nothing. MR. GREGORY: You talk about the perils of default -- as has the, the Fed chief and the president and others -- as a given. And yet look at the polling on this, even among people who are paying close attention in the Gallup poll, 53 percent are in favor of voting against raising the debt ceiling. Do you think that's because there are Republicans like Michele Bachmann and others saying that this administration is really selling, in her words, a "misnomer" that, that we're headed toward default? Where does that come from? Do you think they actually believe that, or you think they're doing that cynically? MR. LEW: I can't explain what motivates people to say, say those things. I can tell you the facts are the facts. If we don't raise the debt ceiling, we won't be able to pay our bills in August. And that has dire consequences. It will, for the first time, mean the United States cannot keep its obligations. It will cascade through the economy. It will mean that people, regular people who are buying homes and, and, and cars will pay higher interest rates. It means that we will put a cloud over the United States that might not go away anytime soon. WASHINGTON (Reuters) - The Senate will likely begin consideration this week of a bipartisan fallback plan to avert an unprecedented U.S. default of its debt, senior Democratic aides said on Sunday. The measure is based on a proposal first offered last week by Senate Republican leader Mitch McConnell. McConnell has been negotiating with Senate Majority Leader Harry Reid on ways to make the proposal more palatable to Democrats. Aides said supporters expect the Democratic-led Senate to pass the legislation, but it's unclear if the Republican-led House of Representatives would provide its needed concurrence. "Right now, things are fluid. The plan isn't yet done," a senior Democratic aide said. As initially proposed by McConnell, the measure would authorize Obama to raise the debt limit by $2.5 trillion -- without any mandatory spending cuts -- provided Obama's fellow Democrats go along with it. Such a move would spare Republicans from having to back an increase in the U.S. borrowing authority, an unpopular move, yet protect them from being blamed for a default. Reid wants up to $1.5 trillion in mandatory spending cuts, along the lines of those identified by a deficit-reduction group headed by Vice President Joe Biden. Staff to McConnell and Reid have also discussed creating a bipartisan committee on long-term deficit reduction, aides said. Democratic aides said while the Senate will likely begin debate on the measure this week, it is unlikely to come up for a vote on passage until next week. "We don't have all the details yet. We are still all talking. Everything is fluid," an aide said. An aide said McConnell and Reid staffers are working under the assumption that White House-led talks with Republican and Democratic leaders won't reach an agreement of their own. "Barring a breakthrough, this is going to be the plan," the aide said. "We are working under the assumption that there won't be a plan." (Reporting by Thomas Ferraro; Editing by Vicki Allen)

Summary: Mitch McConnell and Harry Reid are busily hammering out a debt ceiling Plan B, and are likely to put it to a vote in the coming week, senior Democratic aides tell Reuters. It would be based on McConnell's widely panned plan to allow Democrats to raise the ceiling, but place the blame on them for doing so. Negotiations have centered on how to make Democrats accept the plan. The Senate will also take a symbolic, sure-to-fail vote on a constitutional amendment requiring a balanced budget. Pushback is likely from both sides against any deal. Tom Coburn said today that he is "unlikely to support" any version of McConnell's plan. "It's a great political plan," he said on Face the Nation, according to Roll Call, but it avoids "making the hard choices." White House budget chief Jack Lew meanwhile was optimistic, telling MSNBC that "the leaders in Congress seem to have all agreed that we can't push to a default," and that there is "still time to get something big done."

multi_news

en

Summarize: What Determines Exchange Rates? At times, the exchange rate is erroneously imagined to be an incidental value that can be sustained by the good intentions of government and undermined by the malevolence of greedy speculators. Economic theory holds it to be a value that is far more fundamental. It is the value at which two countries trade goods and services and the value at which investors from one country purchase the assets of another country. As such, it is dependent on the two countries' fundamental macroeconomic conditions, such as its inflation, growth, and saving rates. Thus, it is generally accepted that the value of the exchange rate cannot be predictably altered (for long) unless the country's macroeconomic conditions are modified relative to those of its trading partners. Many view the volatility of floating exchange rates as proof that speculation and irrational behavior, rather than economic fundamentals, drive exchange rate values. Empirical evidence supports the view that changes in exchange rate values are not well correlated with changes in economic data in the short run. But this evidence does not prove that economic theory is wrong. Although floating exchange rate values change frequently, and at times considerably, there are important economic conditions that change frequently in ways that cannot be measured. Factors such as investors' perceptions of future profitability and riskiness cannot be accurately measured, yet changes in these factors can have profound influence on exchange rate values. Economists have had more success at correlating long run exchange rate movements with changes in economic fundamentals. A decision by a government to influence the value of its exchange rate, therefore, is likely to succeed only if its overall macroeconomic conditions are altered. Government does have tools at its disposal to alter aggregate demand in the short run—fiscal and monetary policy. Fiscal policy refers to increasing or decreasing the government's budget surplus (or deficit) in order to increase or decrease the amount of aggregate spending in the economy. Monetary policy refers to increasing or decreasing short-term interest rates through manipulation of the money supply in order to decrease or increase the amount of aggregate spending in the economy. For example, other things being equal, lower interest rates lead to more investment spending, one component of aggregate spending. Furthermore, fiscal and monetary policy influence interest rates differently, and interest rates are the key determinant of the exchange rate. Expansionary fiscal policy is likely to raise interest rates and "crowd out" private investment while expansionary monetary policy, or reducing short-term interest rates, is likely to temporarily lower interest rates. Maintaining a fixed exchange rate requires continuous policy adjustment. Although perhaps theoretically feasible, it would be impossible in practice to operate a timely or precise enough fiscal policy to maintain a fixed exchange rate as long as fiscal policy must be legislated. Thus, maintaining a fixed exchange rate has been delegated to the monetary authority in practice. Intervening in foreign exchange markets directly is equivalent to changing monetary policy if the intervention is "unsterilized." When a central bank sells foreign currency to boost the exchange rate, it takes the domestic currency it receives in exchange out of circulation, decreasing the money supply. Often, it prints new money to replace the domestic currency that has been removed from circulation—referred to as sterilization—but economic theory suggests that when it does so, it negates the intervention's effect on the exchange rate. If a government wishes to alter a floating exchange rate or maintain a fixed exchange rate, it may do so by altering monetary policy but only if it is willing to abandon other macroeconomic goals such as providing stable economic growth, preventing recessions, and maintaining a moderate, stable inflation rate. The magnitude of response of the exchange rate to changes in monetary policy is not likely to be constant or predictable over time, but under most circumstances policy can eventually lead to the desired result if it is truly dedicated to achieving it. As discussed later, problems with exchange rates usually arise when a government's heart is not truly wedded to achieving its stated goal. Floating Exchange Rates The exchange rate arrangement maintained between the United States and all of its major trading partners is known as a floating exchange rate regime. In a floating exchange rate regime, the exchange rate is a price freely determined in the market by supply and demand. The dollar is purchased by foreigners in order to purchase goods or assets from the United States. Likewise, U.S. citizens sell dollars and buy foreign currencies when they wish to purchase goods or assets from foreign countries. The exchange rate is determined by whatever rate clears these markets. Monetary and fiscal policy are not regularly or systematically used to influence the exchange rate. Thus, when the demand for U.S. goods or assets rises relative to the rest of the world, the exchange rate value of the dollar will appreciate. This is necessary to restore balance or equilibrium between the dollar value exported and the dollar value imported. Dollar appreciation accomplishes this through two effects on the United States economy, all else being equal. First, it makes foreign goods cheaper for Americans, which increases the purchasing power of American income. This is known as the terms-of-trade effect. Second, it tends to offset the changes in aggregate demand that first altered the exchange rate by making U.S. exports dearer and foreign imports less expensive. The offset in demand may not be instantaneous or complete, but it helps to make macroeconomic adjustment possible if wages and prices are not completely flexible. When foreigners increase their demand for U.S. goods, aggregate demand in the United States increases. If the United States is in a recession, this increase in aggregate demand would boost growth in the short run. If economic growth in the United States is already robust, it would be inflationary—there would be too many buyers (domestic and foreign) seeking the goods that Americans can produce. Under a floating exchange rate, a substantial part of this increase in U.S. aggregate demand would be offset by the appreciation in the dollar, which would push U.S. exports and the production of U.S. import-competing goods back towards an equilibrium level. By reducing aggregate demand, an appreciating dollar reduces inflationary pressures that might otherwise result. Likewise, if the foreign demand for U.S. assets increased, foreign capital would flow into the United States, lowering interest rates and increasing investment spending and interest-sensitive consumption spending (e.g., automobiles). Absent exchange rate adjustment, this would boost U.S. aggregate demand. But because the greater demand for U.S. assets causes the dollar to appreciate, the demand for U.S. exports and U.S. import-competing goods declines, offsetting the increase in demand caused by the foreign capital inflow. Because floating exchange rates allow for automatic adjustment, they buffer the domestic economy from external changes in international supply and demand. A floating exchange rate also becomes another automatic outlet for internal adjustment. If the economy is growing too rapidly, the exchange rate is likely to appreciate, which helps slow aggregate spending by slowing export growth. While this is unfortunate for exporters, overall it may be preferable to the alternative—higher inflation or a sharp contraction in fiscal or monetary policy to stamp out inflationary pressures. If the economy is in recession with falling income, the exchange rate is likely to depreciate, which will help boost overall growth through export growth even in the absence of domestic recovery. The maintenance of a floating exchange rate does not require support from monetary and fiscal policy. This frees the government to focus monetary and fiscal policy on stabilizing the economy in response to domestic changes in supply and demand. Fiscal and monetary policy usually can be focused on domestic goals, such as maintaining price and output stability, without being constrained by the policy's effect on the exchange rate. The drawback to fiscal and monetary autonomy, of course, is that governments are free to pursue ill-conceived policies if they desire, a particular problem for developing countries historically. Many times, a floating exchange rate is forced to act as an outlet for internal adjustment because poor fiscal and monetary policy have made adjustment necessary, causing stress on the trade sector of the economy. This can be thought of as a political, rather than an economic, drawback to floating exchange rates. How valuable the macroeconomic adjustment mechanism that floating exchange rates provide depends on the economic independence of the country. For countries that are closely tied to others through trade and investment links, the ability to adjust policy independently has little value—whatever is affecting one economy is probably affecting its neighbors as well. For countries like the United States, whose economy is arguably more affected by internal factors than external factors, flexible exchange rates allow significant internal adjustment. Trade is still a relatively small portion of American GDP: exports are equivalent to about 10% of GDP, in comparison to a country like Malaysia or Singapore where exports exceed 100% of GDP. The economic drawback to floating exchange rates is that exchange rate volatility and uncertainty may discourage the growth of trade and international investment. Many developing countries, in particular, have pursued growth strategies that have focused on promoting trade and foreign investment. Exchange rate uncertainty can be thought of as placing a cost on trade and investment, and this cost discourages trade. For example, after an international sale has been negotiated, one party to the transaction will not know what price he will ultimately receive in his currency because upon payment the exchange rate may be higher or lower than when he made the trade. If the exchange rate has depreciated, he will receive lower compensation than he had expected. The cost of this uncertainty can be measured precisely—it is the cost of hedging, that is the cost to the exporter of buying an exchange rate forward contract or futures contract to lock in a future exchange rate today. Hard Pegs and Soft Pegs The alternative to floating exchange rates are exchange rate regimes that fix the value of the exchange rate to that of another country or countries. There are two broad types of fixed exchange rates. "Hard pegs," currency boards and currency unions, are considered first because they are the most stark example of a fixed exchange rate arrangement. The second category considered is fixed exchange rates, in which the link to the other currency or currencies is less direct, making them "soft pegs." Currency Boards or Currency Unions At the opposite end of the spectrum from floating exchange rates are arrangements where a country gives up its exchange rate and monetary freedom entirely by tying itself to a foreign country's currency, what former IMF Deputy Director Stanley Fischer calls "hard pegs." This can be done through a currency board or a currency union. A currency board is a monetary arrangement where a country keeps its own currency, but the central bank cedes all of its power to alter interest rates, and monetary policy is tied to the policy of a foreign country. For example, Hong Kong has a currency board linked to the U.S. dollar. Argentina had a similar arrangement which it abandoned in 2002, during its economic crisis. In Argentina, for every peso of currency in circulation the Argentine currency board held one dollar-denominated asset, and was forbidden from buying and selling domestic assets. Thus, the amount of pesos in circulation could only increase if there was a balance of payment surplus. In effect, the exchange rate at which Argentina competed with foreign goods was set by the United States. Because exchange rate adjustment was not possible, adjustment had to come through prices (i.e., inflation or deflation) instead. Domestically, because the central bank could no longer alter the money supply to change interest rates, the economy could only recover from peaks and valleys of the business cycle through gradual price adjustment. From an economic perspective, a currency union is very similar to a currency board. An example of a currency union is the euro, which has been adopted by 13 members of the European Union. The individual nations in the euro zone have no control over the money supply in their countries. Instead, it is determined by two factors. First, the European Central Bank (ECB) determines the money supply for the entire euro area by targeting short-term interest rates for the euro area as a whole. Second, how much of the euro area's money supply flows to, say, Ireland depends upon Ireland's net monetary transactions with the rest of the euro area. For this second reason, different countries in the euro area have different inflation rates despite the fact that they share a common monetary policy. In a currency union such as the euro arrangement, each member of the euro has a vote in determining monetary policy for the overall euro area. This is the primary difference from a currency board—the country that has adopted a currency board has no say in the setting of monetary policy by the country to which its currency board is tied. The countries of the euro also share in the earnings of the ECB, known as seigniorage, just as they would if they had their own currency. Not all currency unions give all members a say in the determination of monetary policy, however. For instance, when Ecuador, El Salvador, and Panama unilaterally adopted the U.S. dollar as their currency, they gained no influence over the actions and decisions of the Federal Reserve. From a macroeconomic perspective, a unilateral currency adoption and a currency board are indistinguishable. Between these two arrangements, there are only two minor differences of note. First, currency boards earn income on the dollar-denominated assets that they hold (another example of seigniorage) while currency adopters do not. Second, investors may view a currency union as a more permanent commitment than a currency board, particularly after the failure of Argentina's currency board. If this were the case, they would view the risks associated with investment in the former to be lower. Economic Advantages to a Hard Peg The primary economic advantage of a hard peg comes through greater trade with other members of the exchange rate arrangement. The volatility of floating exchange rates places a cost on the export and import-competing sectors of the economy. Greater trade is widely seen to be an engine of growth, particularly among developing countries. In a perfectly competitive world economy without transaction costs, the cost of exchange rate volatility could be very large indeed. For instance, U.S. exporters and domestic firms that compete with importers in 2000 faced one-third higher prices than in 1995 as a result of the (floating) dollar's one-third appreciation against its main trading partners. Until the domestic price level fell by one-third, U.S. producers would be uncompetitive, if all else is equal. (All else was not equal—exports continued to rise in the 1990s despite the dollar's appreciation.) Under a system of fixed exchange rates, U.S. exporters would not have been placed at this price disadvantage, all else being equal. Between small countries, a hard peg is also thought to promote more efficient and competitive markets through lower barriers to entry and greater economies of scale. Hard pegs also encourage international capital flows. The encouragement of international capital flows can enhance a country's welfare in a couple of ways. First, it allows more investment to take place in areas where saving is relatively scarce and rates of return are high, and investment is key to sustainable growth. This makes both the borrower and the investor better off; the former because more investment, and hence growth, is possible than otherwise would be, the latter because they can now enjoy higher rates of return on their investment for a given amount of risk than if limited to home investment. For developing countries, these investment gains can be quite large. Because these countries have much lower capital-labor ratios than the developed world, capital investment can yield relatively high returns for some time if a friendly economic environment is constructed. On the other hand, international capital flows can change rapidly in ways that can be destabilizing to developing countries, as will be discussed below. Weighed against the gains of higher trade and international investment is the loss of the use of fiscal and monetary policy to stabilize the economy. For countries highly integrated with their exchange rate partners, this loss is small. For example, in the euro area, the business cycle of many of the "core" economies (e.g., Germany and the Netherlands, or Belgium and France) have been highly correlated. As long as Belgium does not face separate shocks from France, it does not lose any stabilization capabilities by giving up the ability to set policy independently of France. By sharing a currency, their fiscal and monetary policy can still be adjusted to respond jointly to shared shocks to their economies, even if these shocks are not shared by the rest of the world—the euro is free to adjust against the rest of the world's currencies. Troubles only arise if shocks harm one of these countries, but not its partners in the euro. In that case, there cannot be policy adjustment for that country to compensate for the shock. Political Advantages to a Currency Board or Union The previous explanation described the economic reasons for establishing currency boards or currency unions. But it is probable that the primary reason for establishing them in developing countries is based more on political reasons. As has been shown, these monetary arrangements tie the hands of their country's policymakers. For some countries, this is precisely what their policymakers are trying to achieve—a way to prevent the reinstatement of policies from the "bad old days." The most stark example of the "bad old days" is the hyperinflation that many developing countries experienced. For instance, in 1990, the year before Argentina adopted a currency board, its inflation rate reached 2,314%. Stable growth is impossible when the price mechanism has broken down in this way. The currency board quickly brought the inflation rate in Argentina down to single digits. Whenever a country's inflation rate gets extremely high, it is a reflection of its fiscal policy. Large budget deficits cannot be financed through the sale of debt instruments, so they are instead financed through the printing of money. Thus, a currency board prevents irresponsible fiscal policy by preventing monetary policy from supporting it. Similarly, Ecuador "dollarized" in 2000—adopting the U.S. dollar and largely discontinuing the use of its own currency—at a time of economic crisis with the hope that it would renew investor confidence. Although extremely high inflation had not yet become a problem, events leading up to dollarization appeared to be pointing in that direction. The country's banking system had collapsed, its economy had shrunk by more than 7% in 1999, low oil prices and natural disaster had caused budget financing problems, and it had defaulted on some of its sovereign debt. Investors had become very concerned that inflationary monetary policy would be used to solve its fiscal problems, and dollarization quelled these fears by eliminating that policy option. Economic analysis sheds little light on the choice between floating exchange rates and a currency board arrangement when the decision is motivated by the desire to find a political arrangement that will prevent the pursuit of bad policies. Economic analysis can identify bad policy; it cannot explain why it is pursued or how to prevent its recurrence. A currency board is not the only way to tie the hands of policymakers; various rules and targets have been devised to eliminate policy discretion that could be used with a floating exchange rate. A currency board may be a more final commitment, and hence harder to renege on, than rules and targets, however. Then again, Argentina proved that even currency boards are not permanent. In any case, the political problem of countries monetizing budget deficits seems to be waning. Since 2000, the annual inflation rate in developing countries has averaged about 6%. If current trends continue, in the future there may be fewer countries who find it advantageous to accept the harsh medicine of hard pegs to solve their political shortcomings. Hard pegs are also seen by both proponents and opponents as a means to foster political integration, a topic beyond the scope of this report. This was a primary consideration behind the adoption of the euro. Fixed Exchange Rates In a traditional fixed exchange rate regime, the government has agreed to buy or sell any amount of currency at a predetermined rate. That rate may be linked to one foreign currency or (unlike a currency board) it may be linked to a basket of foreign currencies. In theoretical models, where capital is perfectly mobile and investors consider all countries to be alike, fixed exchange rates would necessarily be functionally equivalent to a currency board. Any attempt to unilaterally influence one's interest rates, through monetary or fiscal policy, would be unsustainable because capital would flow in or out of the country until interest rates had returned to the worldwide level. In reality, results are not quite so stark. There are transaction costs to investment. Investors demand different risk premiums of different countries, and these risk premiums change over time. There is a strong bias among investors worldwide, particularly in developed countries, to keep more of one's wealth invested domestically than economic theory would suggest. Due to these factors, interest rate differentials, which should be theoretically impossible, are abundant. For instance, interest rates in France and Germany should entail similar risks. Thus, anytime French interest rates exceeded German rates, capital should flow from Germany to France until the rates equalized again. Yet the commercial interest reference rate, as measured by the OECD, between these two countries has varied by as much as 1.61 percentage points between 1993 and the adoption of the euro in 1999. As a result, countries with fixed exchange rates have limited freedom to use monetary and fiscal policy to pursue domestic goals without causing their exchange rate to become unsustainable. By contrast, countries that operate currency boards or participate in currency unions have no monetary or fiscal autonomy. For this reason, fixed exchange rates can be thought of as "soft pegs," in contrast to the "hard peg" offered by a currency board or union. But compared to a country with a floating exchange rate, the ability of a country with a fixed exchange rate to pursue domestic goals is highly limited. If a currency became overvalued relative to the country to which it was pegged, then capital would flow out of the country, and the central bank would lose reserves. When reserves are exhausted and the central bank can no longer meet the demand for foreign currency, devaluation ensues, if it has not already occurred before events reach this point. The typical reason for a fixed exchange rate to be abandoned in crisis is due to an unwillingness by the government to abandon domestic goals in favor of defending the exchange rate. Interest rates can almost always be increased to a point where capital no longer flows out of the country, but a great contraction in the economy may accompany those rate increases. It is not uncommon to see interest rates reach triple digits at the height of an exchange rate crisis. Crises ensue because investors do not believe that the government will have the political will to accept the economic hardship required to maintain those interest rates in defense of the currency. Economic Advantages of a Fixed Exchange Rate As with a hard peg, a fixed exchange rate has the advantage of promoting international trade and investment by eliminating exchange rate risk. Because the arrangement may be viewed by market participants as less permanent than a currency board, however, it may generate less trade and investment. As with a hard peg, the drawback of a fixed exchange rate compared to floating exchange rates is that the government has less scope to use monetary and fiscal policy to promote domestic economic stability. Thus, it leaves countries unable to defend themselves against idiosyncratic shocks not shared by the country to which it has fixed its currency. As explained above, this is less of a problem than with a hard peg because imperfect capital mobility does allow for some deviation from the policy of the country or countries to which you are linked. But the shock would need to be temporary in nature because a significant deviation could not last. The scope for the pursuit of domestic goals is greater for countries that fix their exchange rate to a basket of currencies—unlike a hard peg, the country is no longer placed at the mercy of the unique and idiosyncratic policies and shocks of any one foreign country. One method for creating a currency basket is to compose it of the currencies of the country's primary trading partners, particularly if the partner has a hard currency, with shares set in proportion to each country's proportion of trade. If the correlation of the business cycle with each trading partner is proportional to the share of trade with that country, then the potential for idiosyncratic shocks to harm the economy should be considerably reduced when pegged to a basket of currencies. On the down side, baskets do not encourage any more bilateral trade and investment than a floating exchange rate because they reintroduce bilateral exchange rate risk with each trading partner. There is a popular perception that the advantage of a fixed exchange rate is that it allows countries to set their exchange rate below market value in order to boost exports and curb imports. For example, this claim is often leveled against China. Economists would disagree that an artificially low exchange rate is in a country's self interest. Although it has the benefit of boosting a country's trade balance, it also has costs. By making imports more expensive, it reduces consumers' purchasing power. And by distorting market signals, it funnels resources away from their most efficient use. Finally, an undervalued exchange rate confers no permanent trade advantage because it will eventually cause domestic prices to rise, canceling out the price advantage offered by the exchange rate. Political Advantages of a Fixed Exchange Rate In previous decades, it was believed that developing countries with a profligate past could bolster a new commitment to macroeconomic credibility through the use of a fixed exchange rate for two reasons. First, for countries with inflation rates that were previously very high, the maintenance of fixed exchange rates would act as a signal to market participants that inflation was now under control. For example, inflation causes the number of dollars that can be bought with one peso to decline just as it causes the number of apples that can be bought with one peso to decline. Thus, a fixed exchange rate can only be maintained if large inflation differentials are eliminated. Second, a fixed exchange rate was thought to anchor inflationary expectations by providing stable import prices. For a given change in monetary policy, economy theory suggests that inflation will decline faster if people expect lower inflation. After the many crises involving fixed exchange rate regimes in the 1980s and 1990s, this argument has become less persuasive. Unlike a currency board, a fixed exchange rate regime does nothing concrete to tie policymakers' hands and prevent a return to bad macroeconomic policy. Resisting the temptation to finance budget deficits through inflation ultimately depends on political will; if the political will is lacking, then the exchange rate regime will be abandoned, as was the case in many 1980s exchange rate crises. Thus burnt in the past, investors may no longer see a fixed exchange rate as a credible commitment by the government to macroeconomic stability, reducing the benefits of the fixed exchange rate. Furthermore, some currency board proponents claim that this lack of credibility means that investors will "test" the government's commitment to maintaining a soft peg in ways that are costly to the economy. By contrast, they claim that investors will not test a currency board because they have no doubt of the government's commitment. For this reason, many economists who previously recommended fixed exchange rates on the basis of their political merits have shifted in recent years towards support of a hard peg. This has been dubbed the "bipolar view" of exchange rate regimes: growing international capital mobility has made the world economy behave more similarly to what models have suggested. As capital flows become more responsive to interest rate differentials, the ability of "soft peg" fixed exchange rate regimes to simultaneously pursue domestic policy goals and maintain the exchange rate has become untenable. As a result, countries are being pushed toward floating exchange rates (the freedom to pursue domestic goals) or "hard pegs" (policy directed solely toward maintaining the exchange rate). In this view, while "soft pegs" may have been successful in the past, any attempt by a country open to international capital to maintain a soft peg today is likely to end in an exchange rate crisis, as happened to Mexico, the countries of Southeast Asia, Brazil, and Turkey. Empirically, the trend does appear to be moving in this direction. In 1991, 65% of the world's 55 largest economies used "soft peg" exchange rate arrangements; in 1999, the number had fallen to 27%. Although the international trend has been towards greater capital mobility and openness, it should be pointed out that there are still developing countries that are not open to capital flows. The "bipolar view" argument may not hold for these countries: without capital flows reacting to changes in interest rates, these countries may be capable of maintaining a soft peg and an independent monetary policy. For a fixed exchange rate to work, the relative supply and demand for a country's currency must remain stable over time, or the government must adjust the value at which the exchange rate is fixed whenever supply and demand significantly change. In practice, the primary problem with fixed exchange rates has been that countries have faced frequent changes in economic conditions that put pressure on the fixed exchange rate to change, but countries have proven unwilling to change the exchange rate promptly. (Of course, frequent changes undermine many of the economic and political rationales for using fixed exchange rates.) Some countries have faced economic pressures to raise the value of the exchange rate, others to lower it. An undervalued exchange rate can be maintained indefinitely, as long as the country is willing to accumulate foreign exchange reserves. But it may lead to political tensions with trading partners, as has been the case recently between China and the United States. When the exchange rate is overvalued, it frequently results in economic crisis, as will be discussed in the next section. What Have Recent Crises Taught Us About Exchange Rates? The previous discussion summarizes the textbook advantages and disadvantages of different exchange rate regimes. As such, it abstracts and simplifies from many economic issues that may bear directly on real policymaking. In particular, it neglects the possibility that crisis could be caused or transmitted through international goods or capital markets, and the transmission role exchange rates can play in crisis. The remainder of the report will be devoted to trying to glean some general lessons from the international crises of the 1990s, which featured rapidly falling exchange rates and asset prices, international capital flight, and financial unrest, to enrich our understanding of how different exchange rate regimes function. The primary lesson seems to be that fixed exchange rate regimes are prone to crisis, while a crisis caused by international capital movements is extremely improbable under floating regimes. Unlike the crises of the 1980s, most of the countries involved in 1990s crises—particularly Southeast Asia—had relatively good macroeconomic policies in place (e.g., low inflation, balanced budgets, relatively free capital mobility). Thus, these crises cannot be blamed simply on policy errors. Fixed exchange rate regimes are prone to crisis because investors are compelled to remove their money from a country before it devalues. It is similar to a fire in a crowded theater: although everyone easily entered the theater in an orderly fashion, if everyone tries to rush out at once, the doors jam and the fire becomes a catastrophe. Proponents of fixed exchange rate regimes often argue that they can be adjusted if they "get out of line." But the weakness of fixed exchange rate regimes is that when economic fundamentals change in such a way that devaluation becomes necessary, there is no mechanism to devalue except crisis. Even if a government wanted to announce a planned devaluation to avoid crisis, the announcement would likely spur anticipatory capital flight as investors tried to withdraw their investments before the new exchange rate was implemented. Corruption, "crony capitalism," and "greedy speculation" are not needed to explain why fixed exchange rates collapse. The countries forced to devalue during the Asian Crisis (Thailand, Malaysia, Philippines, Indonesia, and South Korea) had very different economic structures and political systems, and were at different stages of economic development, ranging from a per capita GDP of $15,355 in South Korea to $4,111 in Indonesia. What they all had in common was their exchange rate peg to the U.S. dollar. The Asian crisis was instigated by the fact that the appreciating U.S. dollar, to which the crisis countries were fixed, had made their exports less competitive and encouraged imports, particularly compared to China (which had devalued its exchange rate in 1994) and Japan. Investment bubbles, notably in property markets, seemed to be present in all of the crisis countries, although there is no accepted method to identify them even after the fact. Some argue that the bursting of these bubbles played a key role in instigating the crises. Theories for why the bubbles formed include widespread state allocation of capital, poor local financial regulation, and simple misguided exuberance on the part of investors. Whether the bursting of such a bubble could have instigated the crisis under a floating exchange rate is debatable. Some sharp declines in asset prices have sparked serious crises and downturns, as was the case in Japan in the early 1990s. Other times, sharp asset price declines have not caused crisis and have had little lasting effect on the economy, as was the case with the United States in 1987. But what is clear is that an asset bubble and a fixed exchange rate can interact in ways more virulent than their individual parts. To the extent that asset prices would have fallen in Asia to return to their fundamental levels anyway, the presence of a fixed exchange rate ensured that it would happen suddenly because of the "fire in a theater" principle. To the extent that a devaluation would have been necessary anyway, the presence of an asset bubble assured that the outflows would be larger, placing more of a strain on the countries' financial systems. When investors recognize a situation where devaluation becomes likely, even though they may have had no intention of leaving a country otherwise, they have every incentive to remove their money before the devaluation occurs because devaluation makes the local investment worth less in foreign currency. Because the central bank's reserves will always be smaller than liquid capital flows when capital is mobile, devaluation becomes inevitable when investors lose faith in the government's willingness to correct the exchange rate's misalignment. To an extent, the phenomenon then takes on the aspect of a self-fulfilling prophecy. The reason the depreciation of a currency in crisis is typically so dramatic is because at that point investors are no longer leaving because of economic fundamentals, but simply to avoid being the one "standing when the music stops." Notice that in the textbook explanation, a currency depreciation is expected to boost growth through an improved trade balance. In a currency crisis, this does not happen at first, although it does happen eventually, because resources cannot be reallocated towards increased exports quickly enough to compensate for the blow to the economy that comes through the sudden withdrawal of capital. In the Asian crisis, businessmen told of export orders they were unable to fill following devaluation because their credit line had been withdrawn. The shock of the capital outflow is exacerbated by the tendency for banking systems to become unbalanced in fixed exchange rate regimes. When foreigners lending to the banking system start to doubt the sustainability of an exchange rate regime, they tend to shift exchange rate risk from themselves to the banking system in two ways. First, foreign investors denominate their lending in their own currency, so that the financial loss caused by devaluation is borne by the banking system. Before devaluation, a bank's assets might exceed its liabilities. With devaluation, the foreign currency liabilities suddenly multiply in value with the stroke of a pen without any physical change in the economy, and the banks become insolvent. Second, foreign lending to the banking system is done on a short-term basis so that investments can be repatriated before devaluation takes place. This is problematic because most of a bank's investments are longer term. The banks then enter a cycle where the short-term debt is rolled over until crisis strikes, at which point credit lines are cut. Both of these factors lead to a situation where a currency crisis causes a banking crisis, which is a much more significant barrier to economic recovery than the devaluation itself. These two characteristics both tend to be present when lending to developing countries even in good times; the tendencies are accelerated when booms look unsustainable. An exception may have been Brazil, which some economists have suggested recovered so quickly from its devaluation because its banking system had few short-term, foreign currency denominated assets. It is not necessarily illogical for the banking system to accept financing on a short term basis or denominated in foreign currency when credit conditions tighten. If it did not accept all forms of financing available to it, it could face insolvency at worst and a significant contraction in business at best. If the banks believe that the downturn is temporary and the episode will pass without a currency devaluation, then the banks will be able to repay the loans once conditions improve. If devaluation causes them to fail, they may expect the government to bail them out, perhaps explaining their willingness to accept these currency risks. These factors make it clear that once a country enters a currency crisis, there is no policy response that can avoid significant economic dislocation. A policy to lower interest rates to boost aggregate demand and add liquidity to the financial system causes the currency to devalue further, increasing the capital outflow and exacerbating the banking system's insolvency. A policy to raise interest rates in support of the currency exacerbates the economic downturn brought on by crisis by reducing investment demand further. This too can feed through to the banking system and capital markets by bankrupting significant portions of the private sector. And it may not quell the currency crisis. In a textbook analysis, interest rates can always be increased to attract back the capital leaving. In reality, after a certain point higher interest rates increase default risk, perhaps causing more capital flight than lower interest rates would bring. Both the Mexican crisis and the East Asian crisis were exacerbated by contagion effects where crisis spread from country to country in the region. This cannot be explained by an irrational (and degrading) assumption by investors that "all Asians/South Americans are crooks." Rather, it reflects the regional interdependence of these economies. Although there is no a priori evidence that South Korea's currency was overvalued, it became overvalued once its neighbors were forced to devalue. That is because its exports competed with its neighbors, and exports accounted for a large fraction of its GDP. After its neighbors devalued, South Korean exporters, already struggling because the Japanese yen had been depreciating, could no longer offer competitive prices. Simultaneously, it appears that investors' perception of the riskiness of emerging markets in general greatly increased, curtailing lending to South Korea, which placed pressure on interest rates and investment. At this point, the deterioration in economic fundamentals caused the Korean won to become overvalued, and currency crisis spread. One may ask why the Bretton Woods fixed exchange rate system that fixed the currencies of the major western economies from 1945 to 1971 was not prone to crisis (at least before it collapsed). The reason is that capital mobility was largely curtailed under the Bretton Woods system. Without capital mobility, central banks could use their reserves to accommodate small changes in fundamentals and could respond to large changes in fundamentals with a (relatively) orderly devaluation. As long as capital remains mobile—and almost nobody has supported a return to permanent capital controls—the Bretton Woods arrangement cannot be replicated. It was not long after capital controls were removed that the Bretton Woods system experienced a growing number of currency crises in the 1960s and 1970s, leading to its eventual demise. Some economists argue that if short-term, foreign-currency denominated debt is the real culprit in recent crises, then it makes more sense to address the problem directly, rather than through the indirect approach of making it more costly through a floating exchange rate. The problem could be addressed directly through various forms of capital controls, financial regulations, or taxes on capital flows. They argue that capital controls are necessary until financial markets become well enough developed to cope with sudden capital inflows and outflows. Capital controls would also allow countries to operate an independent monetary policy while maintaining the trade-related benefits of a fixed exchange rate, similar to how the Bretton Woods system operated. Yet capital controls deter capital inflows as well as capital outflows, and rapid development is difficult without capital inflows. Capital controls may make crises less likely, but they are also likely to reduce a country's long run sustainable growth rate. That is not to argue that floating exchange rates are stable and predictable, as some economists claimed they would be before their adoption in the 1970s. Rather, it is to argue that their volatility has very little effect on the macroeconomy. For example, the South African rand lost half of its value against the U.S. dollar between 1999 and 2001. Yet GDP growth averaged 2.8% and inflation averaged 5.4% in those years. To be sure, when exchange rates change their value by a significant amount in a few years, exporting and import-competing sectors of the economy suffer. Manufacturing and farming are among those sectors in the United States. But there is very little evidence to suggest that in a well-balanced economy such as the United States, other sectors of the economy cannot pick up the slack when the currency appreciates, especially when monetary policy is applied prudently. The one-third appreciation of the dollar and record trade deficits between 1995 and 2000 did not prevent the U.S. economy from achieving stellar growth and unemployment that at one point dipped below 4%. While floating exchange rates sometimes move by substantial amounts in a couple of years, they do not move by substantial amounts overnight, as happens in fixed exchange rate crises. And that is the key reason why floating exchange rates are not prone to financial and economic crises. Floating and fixed exchange rates both impose costs on economies. Floating exchange rates impose a cost by discouraging trade and investment. Fixed exchange rates impose a cost by limiting policymakers' ability to pursue domestic stabilization, thereby making the economy less stable. But there is a fundamental difference in the types of costs they impose. In most countries, the cost of floating exchange rates is internalized and can be managed through the market in the form of hedging. (Developing countries with undeveloped financial systems may not be able to adequately hedge exchange rate risk, however.) Part of the cost of fixed exchange rates is an externality and cannot be hedged away. In other words, society as a whole bears some of the costs of fixed exchange rate regimes, so that market participants do not take that cost into account in their transactions. The costs that society bears are threefold. First, to the extent that a country faces unique shocks to its economy, it gives up the ability to protect its economy against these shocks. Those involved in international trade and investment do not compensate society at large for the fact that the volatility of aggregate unemployment and inflation has been increased. Second, the fixed exchange rate regime is more prone to crisis, which further increases the probability of high unemployment episodes. Even if floating exchange rates were to lead to lower growth because they dampen the growth of trade and foreign investment, risk averse individuals may prefer that outcome if it leads to fewer crises. Third, in some historical instances, fixed exchange rates have weakened the banking system through their incentives to take on debt that cannot be repaid in the event of devaluation. Of the three factors, the last is the only one that could theoretically be rectified through regulation, although implementing such regulation in practice could be difficult, particularly in the developing world. This is not to argue that fixed exchange rate regimes are never superior to floating regimes. The United States would not be better off with 50 separate currencies for each state even though it would ameliorate regional recessions. When countries economies are interdependent enough, the benefits of fixed exchange rates outweigh the costs: regions experience fewer unique shocks, labor mobility improves, product markets may benefit from greater competition and economies of scale, and capital market integration increases. But few countries meet this criterion. Whether the countries of the euro zone become interdependent enough to make the euro sustainable remains to be seen. At the time the euro was introduced, growth between the "core" (countries like Germany and Italy) and the "periphery" (countries like Ireland and Finland) were widely divergent, although they seem to have narrowed since the euro was introduced. But many developing countries that have adopted (or have considered adopting) fixed exchange rates are not well integrated with the economy to which they are linked (see Appendix ). That is because these countries are looking to link to the world's major "hard" currencies, the U.S. dollar, the euro, the Japanese yen, the British pound, or the Swiss franc. Because they are often choosing to fix their exchange rate to gain credibility (e.g., after an episode of high inflation), only a hard currency would provide that credibility. But because the economies of most developing countries are not closely tied to these hard currency economies, they are likely to face very different economic shocks from the hard currency economy. Therefore, they will not be able to adjust policy in response to the shock because of the fixed exchange, nor will they receive any policy adjustment from the country they are fixed to, because the hard currency country, facing no shock, has no need to adjust its policy. This makes these countries more prone to boom and bust than they would be with a (responsibly run) floating exchange rate. Certainly, Russia and the countries of East Asia and Latin America that were struck by currency crises in the 1990s were not closely enough integrated with the U.S. economy to make a dollar peg sustainable. Of these countries, only Mexico and the Philippines experienced growth that was positively correlated with U.S. growth in the 1990s. Proponents of currency boards argue that they do not suffer the vulnerabilities of traditional fixed exchange rates because devaluation becomes too costly an option for the government to consider. For that reason, they argue, investors have no qualms about the safety of their money, and speculators know they cannot undermine the currency, so they do not try. The example of Argentina's currency board demonstrates why this argument is unpersuasive. In making this argument, currency board proponents are only focusing on the political advantage to a currency board—it makes profligate fiscal and monetary policy impossible. But this is not the only factor that makes economies grow and investors choose them as an investment location. A currency board eliminates currency risk, but it does nothing to eliminate a country's macroeconomic risk, to which investors are just as sensitive. For example, there are good reasons why the overall U.S. economy would not be significantly affected by the dollar's one-third appreciation since 1995, but there is no reason why the Argentine economy would be unaffected. It had not received the large capital inflows or experienced the rapid economic growth that made the dollar's appreciation sustainable—some would argue, desirable—for the United States despite its implication for exporters. Thus, Argentine's exporters and import-competing industries became uncompetitive in the last five years with no countervailing factors to make other sectors of the economy competitive. In fact, developments to the Argentine economy suggest a floating exchange rate would have naturally depreciated in recent years to offset negative factors. The prices of commodities (which are important exports for Argentina) had been falling, foreign investment to developing nations had fallen since the Asian crisis, and Argentina's largest trading partner, Brazil, underwent a significant devaluation in 1998. Although the currency board may have lowered political risk in Argentina, for these reasons, it greatly increased macroeconomic risk, and that is why the currency board collapsed in 2002. In the face of macroeconomic risk and political upheaval, Argentina proved that no currency arrangement is permanent. It is beyond the scope of this report to explore the question of whether developing countries with a profligate economic past can make a credible new start without fixing their exchange rates. Some economists go farther and suggest that in today's globalized economy, fixed exchange rates are no longer viable, and adopting a foreign currency becomes necessary for a country trying to make a new start. In those few cases where a natural currency union partner already exists, a fixed exchange rate offers considerable economic advantages, particularly for a country trying to overcome a profligate past. For all other countries, after considering the experience of recent years, the economic advantages to floating exchange rates seem considerable. Appendix. How Interdependent Are International Economies? The statement that some international economies are naturally suited for floating exchange rate regimes while some economies are naturally suited for a fixed exchange rate with a major trading partner is an uncontroversial statement among economists. It is based on the insights first provided by economist Robert Mundell's model of an optimum currency area, which outlines the criteria that determine under what circumstances a fixed exchange rate would succeed. This model underlines the discussion of advantages and disadvantages presented in the first part of this report. Controversy arises among economists on two points. First, it arises on the political question of how important the political benefits of fixed exchange rates should be, which cannot be addressed by the model. Second, it arises from the fact that the empirical parameters of the optimum currency area model are not well established, with economists disagreeing about how much integration is actually needed for a fixed exchange rate to succeed. This appendix attempts to offer some empirical evidence on the latter question. It approximates a country's interdependence with its largest trading partner based on two key criteria from the optimum currency area model: How closely linked the two countries are through trade, measured as exports to the trading partner as a percentage of total exports in 2005. The degree of correlation between the two countries' business cycles, measured as correlation of economic growth from 1997 to 2006. The results are presented for selected developed countries and areas in Table A-1. Using any specific cutoff point to define two economies as interdependent vs. independent for either measure would be arbitrary, but some countries do not achieve even the bare minimum of interdependence. Negative growth correlation means that, overall, the business cycle in the largest trading partner was typically moving in the opposite direction of the country for any given year in the sample. Typically, this would put pressure on their exchange rates to move in opposite directions as well. Similarly, it would be difficult to argue that the largest trading partner was closely tied to the country economic well-being if it did not receive a large share of the country's exports. By these measures, of the countries in Table A-1, Australia and New Zealand seem poorly suited for a fixed exchange rate on both measures. Both countries are relatively physically isolated and not overly reliant on any particular trading partner. A case could be made for a fixed exchange rate for the other countries in the table; a strong case could be made for Canada and Switzerland. But a closer look at Canada suggests that a successful floating exchange rate may not be incompatible even with a country as closely interdependent with its neighbor as Canada is with the United States. Despite its interdependence, Canada has maintained robust growth and low inflation with a floating exchange rate. Because commodities are a larger percentage of its output than that of the United States, its economy responds to changes in commodity prices differently than the United States does. As a result, its currency has appreciated as commodity prices have risen.

Summary: Congress is generally interested in promoting a stable and prosperous world economy. Stable currency exchange rate regimes are a key component to stable economic growth. This report explains the difference between fixed exchange rates, floating exchange rates, and currency boards/unions, and outlines the advantages and disadvantages of each. Floating exchange rate regimes are market determined; values fluctuate with market conditions. In fixed exchange rate regimes, the central bank is dedicated to using monetary policy to maintain the exchange rate at a predetermined price. In theory, under such an arrangement, a central bank would be unable to use monetary policy to promote any other goal; in practice, there is limited leeway to pursue other goals without disrupting the exchange rate. Currency boards and currency unions, or "hard pegs," are extreme examples of a fixed exchange rate regime where the central bank is truly stripped of all its capabilities other than converting any amount of domestic currency to a foreign currency at a predetermined price. The main economic advantages of floating exchange rates are that they leave the monetary and fiscal authorities free to pursue internal goals-such as full employment, stable growth, and price stability-and exchange rate adjustment often works as an automatic stabilizer to promote those goals. The main economic advantage of fixed exchange rates is that they promote international trade and investment, which can be an important source of growth in the long run, particularly for developing countries. The merits of floating compared to fixed exchange rates for any given country depends on how interdependent that country is with its neighbors. If a country's economy is highly reliant on its neighbors for trade and investment and experiences economic shocks similar to its neighbors', there is little benefit to monetary and fiscal independence, and the country is better off with a fixed exchange rate. If a country experiences unique economic shocks and is economically independent of its neighbors, a floating exchange rate can be a valuable way to promote macroeconomic stability. A political advantage of a currency board or currency union in a country with a profligate past is that it "ties the hands" of the monetary and fiscal authorities, making it harder to finance budget deficits by printing money. Recent experience with economic crisis in Mexico, East Asia, Russia, Brazil, and Turkey suggests that fixed exchange rates can be prone to currency crises that can spill over into wider economic crises. This is a factor not considered in the earlier exchange rate literature, in part because international capital mobility plays a greater role today than it did in the past. These experiences suggest that unless a country has substantial economic interdependence with a neighbor to which it can fix its exchange rate, floating exchange rates may be a better way to promote macroeconomic stability, provided the country is willing to use its monetary and fiscal policy in a disciplined fashion. The collapse of Argentina's currency board in 2002 suggests that such arrangements do not get around the problems with fixed exchange rates, as their proponents claimed. This report does not track legislation and will be updated as events warrant.

gov_report

en

Summarize: CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of U.S. patent application Ser. No. 14/249,032, filed Apr. 9, 2014; which is a continuation of U.S. patent application Ser. No. 13/910,873, filed Jun. 5, 2013, now U.S. Pat. No. 8,728,066; which is a divisional of U.S. patent application Ser. No. 13/277,913, filed Oct. 20, 2011, now U.S. Pat. No. 8,512,326; which claims the benefit of U.S. Provisional Application No. 61/501,106, filed on Jun. 24, 2011 and of U.S. Provisional Application No. 61/531,985, filed on Sep. 7, 2011, the full disclosures of which are incorporated herein by reference. FIELD OF THE INVENTION [0002] The present invention relates systems and methods for the cutting and extraction of uterine fibroid tissue, polyps and other abnormal uterine tissue. BACKGROUND OF THE INVENTION [0003] Uterine fibroids are non-cancerous tumors that develop in the wall of uterus. Such fibroids occur in a large percentage of the female population, with some studies indicating that up to 40 percent of all women have fibroids. Uterine fibroids can grow over time to be several centimeters in diameter and symptoms can include menorrhagia, reproductive dysfunction, pelvic pressure and pain. [0004] One current treatment of fibroids is hysteroscopic resection or myomectomy which involves transcervical access to the uterus with a hysteroscope together with insertion of a cutting instrument through a working channel in the hysteroscope. The cutting instrument may be a mechanical tissue cutter or an electro surgical resection device such as a cutting loop. Mechanical cutting devices are disclosed in U.S. Pat. Nos. 7,226,459; 6,032,673 and 5,730,752 and U.S. Published Patent Appl. 2009/0270898. An electrosurgical cutting device is disclosed in U.S. Pat. No. 5,906,615. [0005] While hysteroscopic resection can be effective in removing uterine fibroids, many commercially available instrument are too large in diameter and thus require anesthesia in an operating room environment. Conventional resectoscopes require cervical dilation to about 9 mm. What is needed is a system that can effectively cut and remove fibroid tissue through a small diameter hysteroscope. SUMMARY OF THE INVENTION [0006] The present invention provides methods for resecting and removing target tissue from a patient&#39;s body, such as fibroids from a uterus. The tissue is cut, captured in a probe, catheter, or other tissue-removal device, and expelled from the capture device by vaporizing a fluid, typically a liquid, adjacent to the captured tissue in order to propel the tissue from the device, typically through an extraction or other lumen present in a body or shaft of the device. Exemplary embodiments of the tissue removal device comprise a reciprocating blade, tubular cutter, or the like, where the blade may be advanced past a cutting window on the device in order to sever a tissue strip and capture the strip within an interior volume or receptacle on the device. The liquid or other expandable fluid is also present in the device, and energy is applied to the fluid in order to cause rapid expansion, e.g. vaporization, in order to propel the severed tissue strip through the extraction lumen. In this way, the dimensions of the extraction lumen can be reduced, particularly in the distal regions of the device where size is of critical importance. [0007] In a first method, according to the present invention, tissue is extracted from an interior of the patient&#39;s body by capturing a tissue volume in a distal portion of an interior passageway of an elongated probe. A fluid located distal to the captured tissue volume is expanded, which proximally propels the tissue volume from the device. The fluid typically comprises a liquid, and the expansion typically comprises a liquid-to-vapor phase transition. In other cases, the fluid might be a gas where the expansion results from very rapid heating. In preferred embodiments, the phase transition is achieved by applying electrical energy in an amount sufficient to vaporize the liquid, typically applying RF current between first and second polarity electrodes, where at least one of the electrodes is disposed on a distal side of the captured tissue volume. [0008] The liquid or other fluid may be provided to a working end of the probe in various ways. Often, the liquid or other fluid is provided from a fluid-filled space in the patient&#39;s body, for example from a distension fluid filled in the cavity to be treated, such as the uterus. Alternatively, the liquid or other fluid may be provided from a remote source through a passageway in the probe. The liquid volume to be vaporized is typically in the range from 0.004 mL to 0.080 mL. [0009] The tissue may be captured in a variety of ways. For example, the tissue may be resected with a blade number or alternatively with an RF electrode. In either case, the resected tissue may then be captured or sequestered within an interior passageway within the blade itself and/or within another portion of the probe. In addition to the propulsion force caused by the vaporizing fluid, the present invention might also rely on applying a negative pressure to a proximal end of the anterior passageway to assist in drawing the tissue in a proximal direction from the extraction lumen. [0010] In a further method according to the present invention, tissue is removed from the interior of a patient&#39;s body by engaging a tubular cutter against the targeted tissue. An RF electrode arrangement on the cutter is energized to electrosurgically cut the tissue, and the same or a different RF electrode is used to vaporize a liquid to apply a positive fluid pressure to a distal surface of the cut tissue. Usually, the same RF electrode arrangement is used to both electrosurgically cut the tissue and to vaporize the liquid. In such instances, the cutter carrying the RF electrode is usually first advanced to electrosurgically cut the tissue and thereafter advanced into the liquid to vaporize the liquid. The liquid is usually present in a chamber or other space having an active electrode at a distal end thereof, and the RF electrode arrangement on the cutter comprises a return electrode. In this way, with the smaller active electrode on the distal side of the tissue, the energy which vaporizes the liquid will be concentrated in the chamber on the distal side of the tissue, thus causing rapid vaporization of the liquid and propulsion of the tissue through the extraction lumen. [0011] In a third method according to the present invention, tissue is cut and extracted from the interior of a patient&#39;s body by reciprocating a cutting member within a tubular cutter body to sever a tissue strip. The severed tissue strip is captured in an extraction lumen of the tubular cutter body, and a phase transition is caused in a fluid distal to the tissue strip to thereby apply a proximally directed expelling or propulsion force to the tissue strip. The phase transition may be caused by applying energy from any one of a variety of energy sources, including an ultrasound transducer, a high-intensity focused ultrasound (HIFU) energy source, a laser energy source, a light or optical energy source, a microwave energy source, a resistive heat source, or the like. Typically, the cutter will carry the energy source, and the energy source is also used to effect cutting of the tissue. In this way the cutter can also carry the energy source into the fluid after the tissue has been cut, and the cutting and vaporization steps can be performed sequentially as the cutter first moves through the tissue and then into the liquid or other fluid to be vaporized. [0012] In a still further method according to the present invention, tissue is cut and extracted by first cutting the tissue with a reciprocating cutting member over an extending stroke and a retracting stroke within a sleeve. The extending stroke cuts and captures tissue which has been drawn through a tissue-receiving window in the sleeve. Vaporization of a liquid distal to the captured tissue is caused by the cutting member while the cutting member is in a transition range between extension and retraction. The tissue is typically captured in the tissue extraction lumen formed at least partially in the cutter member. The cutter member typically carries a cutting electrode, and a second electrode is typically disposed at a distal end of the sleeve. Thus, RF current may be delivered to the cutting electrode and the second electrode in order to both effect cutting of the tissue over the extending stroke of the cutter and to also effect vaporization of the fluid while the cutter is in the transition range. BRIEF DESCRIPTION OF DRAWINGS [0013] FIG. 1 is a plan view of an assembly including a hysteroscope and a tissue-cutting device corresponding to the invention that is inserted through a working channel of the hysteroscope. [0014] FIG. 2 is a schematic perspective view of a fluid management system used for distending the uterus and for assisting in electro surgical tissue cutting and extraction. [0015] FIG. 3 is a cross-sectional view of the shaft of the hysteroscope of FIG. 1 showing various channels therein. [0016] FIG. 4 is a schematic side view of the working end of the electrosurgical tissue-cutting device of FIG. 1 showing an outer sleeve and a reciprocating inner sleeve and an electrode arrangement. [0017] FIG. 5 is a schematic perspective view of the working end of the inner sleeve of FIG. 4 showing its electrode edge. [0018] FIG. 6A is a schematic cut-away view of a portion of outer sleeve, inner RF cutting sleeve and a tissue-receiving window of the outer sleeve. [0019] FIG. 6B is a schematic view of a distal end portion another embodiment of inner RF cutting sleeve. [0020] FIG. 7A is a cross sectional view of the inner RF cutting sleeve of FIG. 6B taken along line 7 A- 7 A of FIG. 6B. [0021] FIG. 7B is another cross sectional view of the inner RF cutting sleeve of FIG. 6B taken along line 7 B- 7 B of FIG. 6B. [0022] FIG. 8 is a schematic view of a distal end portion of another embodiment of inner RF cutting sleeve. [0023] FIG. 9A is a cross sectional view of the RF cutting sleeve of FIG. 8 taken along line 9 A- 9 A of FIG. 8. [0024] FIG. 9B is a cross sectional view of the RF cutting sleeve of FIG. 8 taken along line 9 B- 9 B of FIG. 8. [0025] FIG. 10A is a perspective view of the working end of the tissue-cutting device of FIG. 1 with the reciprocating RF cutting sleeve in a non-extended position. [0026] FIG. 10B is a perspective view of the tissue-cutting device of FIG. 1 with the reciprocating RF cutting sleeve in a partially extended position. [0027] FIG. 10C is a perspective view of the tissue-cutting device of FIG. 1 with the reciprocating RF cutting sleeve in a fully extended position across the tissue-receiving window. [0028] FIG. 11A is a sectional view of the working end of the tissue-cutting device of FIG. 10A with the reciprocating RF cutting sleeve in a non-extended position. [0029] FIG. 1 IB is a sectional view of the working end of FIG. 10B with the reciprocating RF cutting sleeve in a partially extended position. [0030] FIG. 11C is a sectional view of the working end of FIG. 10C with the reciprocating RF cutting sleeve in a fully extended position. [0031] FIG. 12A is an enlarged sectional view of the working end of tissue-cutting device of FIG. 1 IB with the reciprocating RF cutting sleeve in a partially extended position showing the RF field in a first RF mode and plasma cutting of tissue. [0032] FIG. 12B is an enlarged sectional view of the working end of FIG. 11C with the reciprocating RF cutting sleeve almost fully extended and showing the RF fields switching to a second RF mode from a first RF mode shown in FIG. 12A. [0033] FIG. 12C is an enlarged sectional view of the working end of FIG. 11C with the reciprocating RF cutting sleeve again almost fully extended and showing the explosive vaporization of a captured liquid volume to expel cut tissue in the proximal direction. [0034] FIG. 13 is an enlarged perspective view of a portion of the working end of FIG. 12C showing an interior chamber and a fluted projecting element. [0035] FIG. 14 is a sectional view of the working end of FIG. 12C showing an interior chamber and a variation of a projecting element. [0036] FIG. 15 is a sectional view of the working end of FIG. 12C showing an interior chamber and a variation of a projecting element configured to explosively vaporize the captured liquid volume. [0037] FIG. 16A is a perspective view of an alternative working end with a rotational cutter in a window open position. [0038] FIG. 16B is a perspective view of the working end of FIG. 16A with the rotating cutting element in a second position. [0039] FIG. 16C is a view of the working end of FIGS. 16A-16B with the rotating cutting element in a third position. [0040] FIG. 17 is an exploded view of the outer sleeve of the working end of FIGS. 16A-16C showing the mating components comprising a ceramic body and a metal tube. [0041] FIG. 18 is a view of the inner sleeve of the working end of FIGS. 16A-16C de-mated from the outer sleeve. [0042] FIG. 19 is an exploded view of the inner sleeve of FIG. 18 showing the mating components comprising a ceramic body and a metal tube. [0043] FIG. 20A is a cross sectional view of the working end of FIGS. 16A-16C with the rotating inner sleeve in a first position cutting tissue in a first RF mode. [0044] FIG. 20B is a cross sectional view of the working end of FIG. 20A with the rotating inner sleeve in a second window-closed position with a second RF mode vaporizing saline captured in the interior extraction channel. [0045] FIG. 21 is a longitudinal sectional view corresponding to the view of FIG. 20B with the rotating inner sleeve in a window-closed position and with the second RF mode vaporizing saline captured in the interior extraction channel to expel tissue proximally. [0046] FIG. 22 is a view of an alternative embodiment of a metal tube component of an inner sleeve. [0047] FIG. 23 is a view of an alternative embodiment of a metal tube component of an inner sleeve. [0048] FIG. 24 is a perspective view of an alternative probe that is configured to stop the inner rotating sleeve in a particular position. DETAILED DESCRIPTION OF THE INVENTION [0049] FIG. 1 illustrates an assembly that comprises an endoscope 50 used for hysteroscopy together with a tissue-extraction device 100 extending through a working channel 102 of the endoscope. The endoscope or hysteroscope 50 has a handle 104 coupled to an elongated shaft 105 having a diameter of 5 mm to 7 mm. The working channel 102 therein may be round, D-shaped or any other suitable shape. The endoscope shaft 105 is further configured with an optics channel 106 and one or more fluid inflow/outflow channels 108 a, 108 b ( FIG. 3 ) that communicate with valve-connectors 110 a, 110 b configured for coupling to a fluid inflow source 120 thereto, or optionally a negative pressure source 125 ( FIGS. 1-2 ). The fluid inflow source 120 is a component of a fluid management system 126 as is known in the art ( FIG. 2 ) which comprises a fluid container 128 and pump mechanism 130 which pumps fluid through the hysteroscope 50 into the uterine cavity. As can be seen in FIG. 2, the fluid management system 126 further includes the negative pressure source 125 (which can comprise an operating room wall suction source) coupled to the tissue-cutting device 100. The handle 104 of the endoscope includes the angled extension portion 132 with optics to which a videoscopic camera 135 can be operatively coupled. A light source 136 also is coupled to light coupling 138 on the handle of the hysteroscope 50. The working channel 102 of the hysteroscope is configured for insertion and manipulation of the tissue-cutting and extracting device 100, for example to treat and remove fibroid tissue. In one embodiment, the hysteroscope shaft 105 has an axial length of 21 cm, and can comprise a 0° scope, or 15° to 30° scope. [0050] Still referring to FIG. 1, the tissue-cutting device 100 has a highly elongated shaft assembly 140 configured to extend through the working channel 102 in the hysteroscope. A handle 142 of the tissue-cutting device 100 is adapted for manipulating the electro surgical working end 145 of the device. In use, the handle 142 can be manipulated both rotationally and axially, for example, to orient the working end 145 to cut targeted fibroid tissue. The tissue-cutting device 100 has subsystems coupled to its handle 142 to enable electrosurgical cutting of targeted tissue. A radiofrequency generator or RF source 150 and controller 155 are coupled to at least one RF electrode carried by the working end 145 as will be described in detail below. In one embodiment shown in FIG. 1, an electrical cable 156 and negative pressure source 125 are operatively coupled to a connector 158 in handle 142. The electrical cable couples the RF source 150 to the electrosurgical working end 145. The negative pressure source 125 communicates with a tissue-extraction channel 160 in the shaft assembly 140 of the tissue extraction device 100 ( FIG. 4 ). [0051] FIG. 1 further illustrates a seal housing 162 that carries a flexible seal 164 carried by the hysteroscope handle 104 for sealing the shaft 140 of the tissue-cutting device 100 in the working channel 102 to prevent distending fluid from escaping from a uterine cavity. [0052] In one embodiment as shown in FIG. 1, the handle 142 of tissue-cutting device 100 includes a motor drive 165 for reciprocating or otherwise moving a cutting component of the electrosurgical working end 145 as will be described below. The handle 142 optionally includes one or more actuator buttons 166 for actuating the device. In another embodiment, a footswitch can be used to operate the device. In one embodiment, the system includes a switch or control mechanism to provide a plurality of reciprocation speeds, for example 1 Hz, 2 Hz, 3 Hz, 4 Hz and up to 8 Hz. Further, the system can include a mechanism for moving and locking the reciprocating cutting sleeve in a non-extended position and in an extended position. Further, the system can include a mechanism for actuating a single reciprocating stroke. [0053] Referring to FIGS. 1 and 4, an electrosurgical tissue-cutting device has an elongate shaft assembly 140 extending about longitudinal axis 168 comprising an exterior or first outer sleeve 170 with passageway or lumen 172 therein that accommodates a second or inner sleeve [0000] 175 that can reciprocate (and optionally rotate or oscillate) in lumen 172 to cut tissue as is known in that art of such tubular cutters. In one embodiment, the tissue-receiving window 176 in the outer sleeve 170 has an axial length ranging between 10 mm and 30 mm and extends in a radial angle about outer sleeve 170 from about 45° to 210° relative to axis 168 of the sleeve. The outer and inner sleeves 170 and 175 can comprise a thin-wall stainless steel material and function as opposing polarity electrodes as will be described in detail below. FIGS. 6A-8 illustrate insulative layers carried by the outer and inner sleeves 170 and 175 to limit, control and/or prevent unwanted electrical current flows between certain portions of the sleeve. In one embodiment, a stainless steel outer sleeve 170 has an O.D. of 0.143″ with an I.D. of 0.133″ and with an inner insulative layer (described below) the sleeve has a nominal I.D. of 0.125″. In this embodiment, the stainless steel inner sleeve 175 has an O.D. of 0.120″ with an I.D. of 0.112″. The inner sleeve 175 with an outer insulative layer has a nominal O.D. of about 0.123″ to 0.124″ to reciprocate in lumen 172. In other embodiments, outer and or inner sleeves can be fabricated of metal, plastic, ceramic of a combination thereof. The cross-section of the sleeves can be round, oval or any other suitable shape. [0054] As can be seen in FIG. 4, the distal end 177 of inner sleeve 175 comprises a first polarity electrode with distal cutting electrode edge 180 about which plasma can be generated. The electrode edge 180 also can be described as an active electrode during tissue cutting since the electrode edge 180 then has a substantially smaller surface area than the opposing polarity or return electrode. In one embodiment in FIG. 4, the exposed surfaces of outer sleeve 170 comprises the second polarity electrode 185, which thus can be described as the return electrode since during use such an electrode surface has a substantially larger surface area compared to the functionally exposed surface area of the active electrode edge 180. [0055] In one aspect of the invention, the inner sleeve or cutting sleeve 175 has an interior tissue extraction lumen 160 with first and second interior diameters that are adapted to electrosurgically cut tissue volumes rapidly—and thereafter consistently extract the cut tissue strips through the highly elongated lumen 160 without clogging. Now referring to FIGS. 5 and 6A, it can be seen that the inner sleeve 175 has a first diameter portion 190 A that extends from the handle 142 ( FIG. 1 ) to a distal region 192 of the sleeve 175 wherein the tissue extraction lumen transitions to a smaller second diameter lumen 190 B with a reduced diameter indicated at B which is defined by the electrode sleeve element 195 that provides cutting electrode edge 180. The axial length C of the reduced cross-section lumen 190 B can range from about 2 mm to 20 mm. In one embodiment, the first diameter A is 0.112″ and the second reduced diameter B is 0.100″. As shown in FIG. 5, the inner sleeve 175 can be an electrically conductive stainless steel and the reduced diameter electrode portion also can comprise a stainless steel electrode sleeve element 195 that is welded in place by weld 196 ( FIG. 6A ). In another alternative embodiment, the electrode and reduced diameter electrode sleeve element 195 comprises a tungsten tube that can be press fit into the distal end 198 of inner sleeve 175. FIGS. 5 and 6A further illustrates the interfacing insulation layers 202 and 204 carried by the first and second sleeves 170, 175, respectively. In FIG. 6A, the outer sleeve 170 is lined with a thin-wall insulative material 200, such as PFA, or another material described below. Similarly, the inner sleeve 175 has an exterior insulative layer 202. These coating materials can be lubricious as well as electrically insulative to reduce friction during reciprocation of the inner sleeve 175. [0056] The insulative layers 200 and 202 described above can comprise a lubricious, hydrophobic or hydrophilic polymeric material. For example, the material can comprise a bio-compatible material such as PFA, TEFLON®, polytetrafluroethylene (PTFE), FEP (Fluorinated ethylenepropylene), polyethylene, polyamide, ECTFE (Ethylenechlorotrifluoro-ethylene), ETFE, PVDF, polyvinyl chloride or silicone. [0057] Now turning to FIG. 6B, another variation of inner sleeve 175 is illustrated in a schematic view together with a tissue volume being resected with the plasma electrode edge 180. In this embodiment, as in other embodiments in this disclosure, the RF source operates at selected operational parameters to create a plasma around the electrode edge 180 of electrode sleeve 195 as is known in the art. Thus, the plasma generated at electrode edge 180 can cut and ablate a path P in the tissue 220, and is suited for cutting fibroid tissue and other abnormal uterine tissue. In FIG. 6B, the distal portion of the cutting sleeve 175 includes a ceramic collar 222 which is adjacent the distal edge 180 of the electrode sleeve 195. The ceramic 222 collar functions to confine plasma formation about the distal electrode edge 180 and functions further to prevent plasma from contacting and damaging the polymer insulative layer 202 on the cutting sleeve 175 during operation. In one aspect of the invention, the path P cut in the tissue 220 with the plasma at electrode edge 180 provides a path P having an ablated width indicated at W, wherein such path width W is substantially wide due to tissue vaporization. This removal and vaporization of tissue in path P is substantially different than the effect of cutting similar tissue with a sharp blade edge, as in various prior art devices. A sharp blade edge can divide tissue (without cauterization) but applies mechanical force to the tissue and may prevent a large cross section slug of tissue from being cut. In contrast, the plasma at the electrode edge 180 can vaporize a path P in tissue without applying any substantial force on the tissue to thus cut larger cross sections of slugs or strips of tissue. Further, the plasma cutting effect reduces the cross section of tissue strip 225 received in the reduced cross-section region 190 B of tissue-extraction lumen 160. FIG. 6B depicts a tissue strip 225 entering the reduced cross-section region 190 B, wherein the tissue strip 225 has a smaller cross-section than the lumen due to the vaporization of tissue. Further, the cross section of tissue 225 as it enters the larger cross-section lumen 190 A results in even greater free space 196 around the tissue strip 225. Thus, the resection of tissue with the plasma electrode edge 180, together with the lumen transition from the smaller cross-section ( 190 B) to the larger cross-section ( 190 A) of the tissue-extraction lumen 160 can significantly reduce or eliminate the potential for successive resected tissue strips 225 to clog the lumen. Prior art resection devices with such small diameter tissue-extraction lumen typically have problems with tissue clogging. [0058] In another aspect of the invention, the negative pressure source 225 coupled to the proximal end of tissue-extraction lumen 160 (see FIGS. 1 and 4 ) also assists in aspirating and moving tissue strips 225 in the proximal direction to a collection reservoir (not shown) outside the handle 142 of the device. [0059] FIGS. 7A-7B illustrate the change in lumen diameter of cutting sleeve 175 of FIG. 6B. FIG. 8 illustrates the distal end of a variation of cutting sleeve 175 ′ which is configured with an electrode cutting element 195 ′ that is partially tubular in contrast to the previously described tubular electrode element 195 ( FIGS. 5 and 6A ). FIGS. 9A-9B again illustrate the change in cross-section of the tissue-extraction lumen between reduced cross-section region 190 B′ and the increased cross-section region 190 A′ of the cutting sleeve 175 ′ of FIG. 8. Thus, the functionality remains the same whether the cutting electrode element 195 ′ is tubular or partly tubular. In FIG. 8A, the ceramic collar 222 ′ is shown, in one variation, as extending only partially around sleeve 175 ′ to cooperate with the radial angle of cutting electrode element 195 ′. Further, the variation of FIG. 8 illustrates that the ceramic collar 222 ′ has a larger outside diameter than insulative layer 202. Thus, friction may be reduced since the short axial length of the ceramic collar 222 ′ interfaces and slides against the interfacing insulative layer 200 about the inner surface of lumen 172 of outer sleeve 170. [0060] In general, one aspect of the invention comprises a tissue cutting and extracting device ( FIGS. 10A-11C ) that includes first and second concentric sleeves having an axis and wherein the second (inner) sleeve 175 has an axially-extending tissue-extraction lumen therein, and wherein the second sleeve 175 is moveable between axially non-extended and extended positions relative to a tissue-receiving window 176 in first sleeve 170 to resect tissue, and wherein the tissue extraction lumen 160 has first and second cross-sections. The second sleeve 175 has a distal end configured as a plasma electrode edge 180 to resect tissue disposed in tissue-receiving window 176 of the first sleeve 170. Further, the distal end of the second sleeve, and more particularly, the electrode edge 180 is configured for plasma ablation of a substantially wide path in the tissue. In general, the tissue-extraction device is configured with a tissue extraction lumen 160 having a distal end portion with a reduced cross-section that is smaller than a cross-section of medial and proximal portions of the lumen 160. [0061] In one aspect of the invention, referring to FIGS. 7A-7B and 9A-9B, the tissue-extraction lumen 160 has a reduced cross-sectional area in lumen region 190 B proximate the plasma cutting tip or electrode edge 180 wherein said reduced cross section is less than 95%, 90%, 85% or 80% of the cross sectional area of medial and proximal portions 190 A of the tissue-extraction lumen, and wherein the axial length of the tissue-extraction lumen is at least 10 cm, 20 cm, 30 cm or 40 cm. In one embodiment of tissue-cutting device 100 for hysteroscopic fibroid cutting and extraction ( FIG. 1 ), the shaft assembly 140 of the tissue-cutting device is 35 cm in length. [0062] FIGS. 10A-10C illustrate the working end 145 of the tissue-cutting device 100 with the reciprocating cutting sleeve or inner sleeve 175 in three different axial positions relative to the tissue receiving window 176 in outer sleeve 170. In FIG. 10 A, the cutting sleeve 175 is shown in a retracted or non-extended position in which the sleeve 175 is at it proximal limit of motion and is prepared to advance distally to an extended position to thereby electrosurgically cut tissue positioned in and/or suctioned into window 176. FIG. 10B shows the cutting sleeve 175 moved and advanced distally to a partially advanced or medial position relative to tissue cutting window 176. FIG. 10C illustrates the cutting sleeve 175 fully advanced and extended to the distal limit of its motion wherein the plasma cutting electrode 180 has extended past the distal end 226 of tissue-receiving window 176 at which moment the resected tissue strip 225 in excised from tissue volume 220 and captured in reduced cross-sectional lumen region 190 B. [0063] Now referring to FIGS. 10A-10C, FIGS. 11A-11C and FIGS. 12A-12C, another aspect of the invention comprises “tissue displacement” mechanisms provided by multiple elements and processes to “displace” and move tissue strips 225 ( FIG. 12A ) in the proximal direction in lumen 160 of cutting sleeve 175 to thus ensure that tissue does not clog the lumen of the inner sleeve 175. As can be seen in FIG. 10A and the enlarged views of FIGS. 11A-11C, one tissue displacement mechanism comprises a projecting element 230 that extends proximally from distal tip 232 which is fixedly attached to outer sleeve 170. The projecting element 230 extends proximally along central axis 168 in a distal chamber 240 defined by outer sleeve 170 and distal tip 232. In one embodiment depicted in FIG. 11 A, the shaft-like projecting element 230, in a first functional aspect, comprises a mechanical pusher that functions to push a captured tissue strip 225 proximally from the small cross-section lumen 190 B of cutting sleeve 175 ( FIG. 12A ) as the cutting sleeve 175 moves to its fully advanced or extended position. [0064] In a second functional aspect, the chamber 240 in the distal end of sleeve 170 is configured to capture a volume of saline distending fluid 244 ( FIG. 12A ) from the working space, and wherein the existing RF electrodes of the working end 145 are further configured to explosively vaporize the captured fluid 244 to generate proximally-directed forces on tissue strips 225 resected and disposed in lumen 160 of the cutting sleeve 175 ( FIGS. 12B and 12C ). Both of these functional elements and processes (tissue displacement mechanisms) can apply a substantial mechanical force on the captured tissue strips 225 by means of the explosive vaporization of liquid in chamber 240 and can function to move tissue strips 225 in the proximal direction in the tissue-extraction lumen 160. It has been found that using the combination of multiple functional elements and processes can virtually eliminate the potential for tissue clogging the tissue extraction lumen 160. [0065] More particularly, FIGS. 12A-12C illustrate the functional aspects of the tissue displacement mechanisms and the subsequent explosive vaporization of fluid captured in chamber 240. In FIG. 12A, the reciprocating cutting sleeve 175 is shown in a medial position advancing distally wherein plasma at the cutting electrode edge 180 is cutting a tissue strip 225 that is disposed within lumen 160 of the cutting sleeve 175. In FIG. 12A-12C, it can be seen that the system operates in first and second electrosurgical modes corresponding to the reciprocation and axial range of motion of cutting sleeve 175 relative to the tissue-receiving window 176. As used herein, the term “electrosurgical mode” refers to which electrode of the two opposing polarity electrodes functions as an “active electrode” and which electrode functions as a “return electrode”. The terms “active electrode” and “return electrode” are used in accordance with convention in the art—wherein an active electrode has a smaller surface area than the return electrode which thus focuses RF energy density about such an active electrode. In the working end 145 of FIGS. 10A-11C, the cutting electrode element 195 and its cutting electrode edge 180 must comprise the active electrode to focus energy about the electrode to generate the plasma for tissue cutting. Such a high-intensity, energetic plasma at the electrode edge 180 is needed throughout stroke X indicated in FIG. 12A-12B to cut tissue. The first mode occurs over an axial length of travel of inner cutting sleeve 175 as it crosses the tissue-receiving window 176, at which time the entire exterior surface of outer sleeve 170 comprises the return electrode indicated at 185. The electrical fields EF of the first RF mode are indicated generally in FIG. 12A. [0066] FIG. 12 B illustrates the moment in time at which the distal advancement or extension of inner cutting sleeve 175 entirely crosses the tissue-receiving window 176 ( FIG. 12A ). At this time, the electrode sleeve 195 and its electrode edge 180 are confined within the mostly insulated-wall chamber 240 defined by the outer sleeve 170 and distal tip 232. At this moment, the system is configured to switch to the second RF mode in which the electric fields EF switch from those described previously in the first RF mode. As can be seen in FIG. 12B, in this second mode, the limited interior surface area 250 ( FIG. 12C ) of distal tip 232 that interfaces chamber 240 functions as an active electrode and the distal end portion of cutting sleeve 175 exposed to chamber 240 acts as a return electrode. In this mode, very high energy densities occur about surface 250 and such a contained electric field EF can explosively and instantly vaporize the fluid 244 captured in chamber 240. The expansion of water vapor can be dramatic and can thus apply tremendous mechanical forces and fluid pressure on the tissue strip 225 to move the tissue strip in the proximal direction in the tissue extraction lumen 160. FIG. 12C illustrates such explosive or expansive vaporization of the distention fluid 244 captured in chamber 240 and further shows the tissue strip 225 being expelled in the proximal direction the lumen 160 of inner cutting sleeve 175. [0067] FIG. 14 shows the relative surface areas of the active and return electrodes at the extended range of motion of the cutting sleeve 175, again illustrating that the surface area of the non-insulated distal end surface 250 is small compared to surface 255 of electrode sleeve which comprises the return electrode. [0068] Still referring to FIGS. 12A-12C, it has been found that a single power setting on the RF source 150 and controller 155 can be configured both (i) to create plasma at the electrode cutting edge 180 of electrode sleeve 195 to cut tissue in the first mode, and (ii) to explosively vaporize the captured distention fluid 244 in the second mode. Further, it has been found that the system can function with RF mode-switching automatically at suitable reciprocation rates ranging from 0.5 cycles per second to 8 or 10 cycles per second. In bench testing, it has been found that the tissue-cutting device described above can cut and extract tissue at the rate of from 4 grams/min to 8 grams/min without any potential for tissue strips 225 clogging the tissue-extraction lumen 160. In these embodiments, the negative pressure source 125 also is coupled to the tissue-extraction lumen 160 to assist in applying forces for tissue extraction. [0069] Of particular interest, the fluid-capture chamber 240 defined by sleeve 170 and distal tip 232 can be designed to have a selected volume, exposed electrode surface area, length and geometry to optimize the application of expelling forces to resected tissue strips 225. In one embodiment, the diameter of the chamber is 3.175 mm and the length is 5.0 mm which taking into account the projecting element 230, provided a captured fluid volume of approximately 0.040 mL. In other variations, the captured fluid volume can range from 0.004 mL to 0.080 mL. [0070] In one example, a chamber 240 with a captured liquid volume of 0.040 mL together with 100% conversion efficiency in and instantaneous vaporization would require 103 Joules to heat the liquid from room temperature to water vapor. In operation, since a Joule is a W*s, and the system reciprocate at 3 Hz, the power required would be on the order of 311 W for full, instantaneous conversion to water vapor. A corresponding theoretical expansion of 1700× would occur in the phase transition, which would results in up to 25,000 psi instantaneously (14.7 psi×1700), although due to losses in efficiency and non-instantaneous expansion, the actual pressures would be much less. In any event, the pressures are substantial and can apply significant expelling forces to the captured tissue strips 225. [0071] Referring to FIG. 12A, the interior chamber 240 can have an axial length from about 0.5 mm to 10 mm to capture a liquid volume ranging from about 0.004 mL 0.01 mL. It can be understood in FIG. 12A, that the interior wall of chamber 240 has an insulator layer 200 which thus limits the electrode surface area 250 exposed to chamber 240. In one embodiment, the distal tip 232 is stainless steel and is welded to outer sleeve 170. The post element 248 is welded to tip 232 or machined as a feature thereof. The projecting element 230 in this embodiment is a non-conductive ceramic. [0072] FIG. 13 shows the cross-section of the ceramic projecting element 230 which may be fluted, and which in one embodiment has three flute elements 260 and three corresponding axial grooves 262 in its surface. Any number of flutes, channels or the like is possible, for example from two to about 20. The fluted design increases the available cross-sectional area at the proximal end of the projecting element 230 to push the tissue strip 225, while at the same time the three grooves 262 permit the proximally-directed jetting of water vapor to impact the tissue exposed to the grooves 262. In one embodiment, the axial length D ( FIG. 12A ) of the projecting element 230 is configured to push tissue entirely out of the reduced cross-sectional region 190 B of the electrode sleeve element 195. In another embodiment, the volume of the chamber 240 is configured to capture liquid that when explosively vaporized provides a gas (water vapor) volume sufficient to expand into and occupy at least the volume defined by a 10% of the total length of extraction channel 160 in the device, usually at least 20% of the extraction channel 160, often at least 40% of the extraction channel 160, sometimes at least 60% of the extraction channel 160, other times at least 80% of the extraction channel 160, and sometimes at least 100% of the extraction channel 160. [0073] As can be understood from FIGS. 12A to 12C, the distending fluid 244 in the working space replenishes the captured fluid in chamber 240 as the cutting sleeve 175 moves in the proximal direction or towards its non-extended position. Thus, when the cutting sleeve 175 again moves in the distal direction to cut tissue, the interior chamber 240 is filled with fluid 244 which is then again contained and is then available for explosive vaporization as described above when the cutting sleeve 175 closes the tissue-receiving window 176. In another embodiment, a one-way valve can be provided in the distal tip 232 to draw fluid directly into interior chamber 240 without the need for fluid to migrate through window 176. [0074] FIG. 15 illustrates another variation in which the active electrode surface area 250 ′ in the second mode comprises a projecting element 230 with conductive regions and non-conductive regions 260 which can have the effect of distributing the focused RF energy delivery over a plurality of discrete regions each in contact with the captured fluid 244. This configuration can more efficiently vaporize the captured fluid volume in chamber 240. In one embodiment, the conductive regions 250 ′ can comprise metal discs or washers on post 248. In other variation (not shown) the conductive regions 250 ′ can comprise holes, ports or pores in a ceramic material 260 fixed over an electrically conductive post 248. [0075] In another embodiment, the RF source 150 and controller 155 can be programmed to modulate energy delivery parameters during stroke X and stroke Y in FIGS. 12A-12C to provide the optimal energy (i) for plasma cutting with electrode edge 180, and (ii) for explosively vaporizing the captured fluid in chamber 240. [0076] FIGS. 16A-16C illustrate another embodiment RF cutting probe 700 with working end 702 comprising a tubular cutter adapted for electrosurgical cutting and extracting targeted tissue from the interior of a patient&#39;s body. However, in this embodiment, the inner cutting sleeve is configured to rotate instead of reciprocate as in the previously-described embodiments. [0077] Referring to FIG. 16A, the outer sleeve 705 comprises a metal tubular member 708 that extends from a handle (not shown) to a working end 702 that again carries a distal dielectric body 710 defining a window 712 therein. The inner second sleeve or cutting sleeve 715 comprises a metal tubular member 718 that carries a distal dielectric body 720 with a windowed side 724 that is adapted to cooperate with window 712 in the outer sleeve 705. [0078] FIGS. 16B-16C show the working end 702 of probe 700 with the rotating cutting sleeve 715 and RF electrode edge 725 in two different rotational positions with respect to outer sleeve 705 and window 712. In FIG. 16B, the inner sleeve 715 is rotated approximately 90° relative to the outer sleeve 705. In FIG. 16C, the inner sleeve 715 is rotated 180° to a position relative to inner sleeve 715 to effectively close the window 712 defined by the outer sleeve 705. It can easily be understood how rotation of electrode edge 725 thus can cut tissue during rotation and capture the tissue in the window-closed position within the tissue-receiving lumen 730 of the probe. [0079] In this embodiment of FIGS. 16A-16C, the RF electrode edge 725 of the inner sleeve 715 comprises a first polarity electrode. The exterior surface 732 of the outer sleeve 705 comprises a second polarity electrode as described in previous embodiments. As can be understood from FIGS. 16A-16C, it is critical that the first and second polarity electrode surfaces ( 725 and 732 ) are spaced apart by a predetermined dimension throughout the rotation of inner sleeve 715 relative to outer sleeve 705. In one aspect the invention, the distal ends of the inner and outer sleeves comprise ceramic bodies 710 and 720 with an interface 740 therebetween. In other words, the ceramic bodies 710 and 720 rotate about interface 740 and the bodies provide exact electrode spacing ES between the first and second polarity electrodes 725 and 732. [0080] Now referring to FIG. 17, it can be seen how the outer sleeve 705 comprises as an assembly between the tubular metal sleeve 708 and the dielectric body 710, which in this variation can be a ceramic such as zirconium. In FIG. 17, it can be seen that the ceramic body 710 has a thin wall 742 which can range in thickness from about 0.003″ and 0.010″ wherein the ceramic extends 360° around window 712. Ceramic body 710 can thus be slidably inserted into and bonded to bore 728 in metal sleeve 708. [0081] Now turning to FIG. 18, the distal end of inner sleeve 715 is shown de-mated from the outer sleeve assembly 705 (see FIG. 16A ). The tubular metal sleeve 718 of FIG. 18 is fabricated to allow insertion of the ceramic body 720 which supports the electrode edge 725 and provides a rotational bearing surface about the interface 740 (see FIG. 16A ). FIG. 19 shows an exploded view of the inner sleeve assembly of FIG. 18. In FIG. 19, it can be seen that ceramic body 720 has a hemispherical cross-sectional shape and includes an elongated slots 744 for receiving and supporting an electrode edge 725. FIG. 19 further shows metal sleeve 718 without ceramic body 720 wherein the electrode edge 725 is cut from a rounded end sleeve 718. It can be understood that the slot 744 can receive ceramic body 720 and thus the electrode edge 725 extends in a loop and under rotation will have a leading edge 745 and a trailing edge 745 ′ depending on the direction of rotation. As used herein, the term ‘leading edge’ refers to the electrode edge 725 extending around the distal end of the sleeve 715 to its centerline on its rotational axis. [0082] In one aspect of the invention, the tissue cutting probe 700 comprises an outer sleeve 705 and an inner sleeve 715 that is rotatable to provide window-open and window-closed positions and wherein the distal ends of the first and second sleeves 705, 715 include ceramic bodies 710, 720 that provide surfaces on either side of a rotational interface 740. Further, the first and second sleeves provide ceramic bodies 710, 720 that contact one another on either side of the rotational interface 740 and thus provide a predetermined electrode spacing ES ( FIG. 16A ). In one variation, the wall thickness of the ceramic body 710 is from 0.003″ to 0.004″. Likewise, the wall thickness of ceramic body 720 can be from 0.003″ to 0.004″. Thus, the radial dimension between the first and second polarity electrodes at a minimum in this variation is 0.006″. In another variation in which the inner sleeve 715 carries an outer polymeric dielectric layer which can be 0.001″ in thickness to thus provide an electrode spacing dimension ES of 0.004″. In other variations having a larger diameter, the dimension between the first and second polarity electrodes can range up to 0.030″. In general, the scope of the invention includes providing a rotational tubular cutter with bi-polar electrodes spaced apart between 0.004″ inches and 0.030″ inches wherein the cutting sleeve 715 rotates about an interface 740 having dielectric materials on either side thereof. [0083] In the embodiment shown in FIGS. 16A-16C, the length of the window can range from about 5 mm to 30 mm. The diameter of the probe working end can range from about 3 mm to 6 mm or more. The rotational speed of the inner sleeve can range from 100 rpm to 5,000 rpm. In one embodiment, a rotation ranging from about 200 rpm to 500 rpm cut tissue efficiently and allowed for effective tissue extraction as described below. [0084] In another aspect of the invention, referring to FIGS. 17, 20A and 20B, it can be seen that an opening 748 is provided in ceramic body 710 which provides exposure through the ceramic body 710 to metal sleeve 708 which comprises the first polarity electrode when assembled. Thus, the metal sleeve provides an interior electrode surface 750 that is exposed to interior chamber 730. It can be understood that in this variation, the working end 702 can function in two RF modes as described in the previous reciprocating probe embodiments (see FIGS. 12A-12C ). In the first RF mode, the exterior surface 732 of outer sleeve 705 functions as a first polarity electrode in the interval when the inner sleeve 715 and its second polarity electrode edge 725 rotates from the window-open position of FIG. 16A toward the window-closed position of FIG. 16B. FIG. 20A depicts this interval of rotation, wherein it can be seen that the first RF mode operates for approximately 180° of rotation of the inner cutting sleeve 715. In this position depicted in FIG. 20A, the leading edge 745 and trailing edge 745 ′ of electrode edge 725 are exposed to the open window 712 and electric fields EF extend to the first polarity electrode surface 732 about the exterior of the probe and plasma is formed at leading edge 745 to cut tissue. [0085] The second RF mode is shown in FIG. 20B, wherein the inner sleeve 715 rotates to the window-closed position and the probe switches instantly to such a second RF mode since the electrode edge 725 is exposed only to the tissue-receiving lumen 730. It can be understood that the second RF mode operates only when the window 712 is closed as in FIGS. 16C and 20B which causes the instant explosive vaporization of captured saline in the lumen 730. In FIG. 20B, it can be seen that the electrode edge 725 is exposed only to the interior of lumen 730 and electric fields EF extend between the leading and trailing electrode edges ( 745 and 745 ′) to the exposed electrode surface 750 to thus cause the explosive vaporization of captured saline. The vaporization occurs instantly within limited degrees of rotation of the inner sleeve, e.g., 5° to 20° of rotation, upon closing the window 712 to thereby expel the resected tissue in the proximal direction as described previously. It has been found that saline captured in the interior channel 730 can be distal to the resected tissue or adjacent to the resected tissue in the lumen and the fluid expansion in the liquid-to-vapor transition will instantly expel the resected tissue outwardly or proximally in lumen 730. [0086] FIG. 21 is a longitudinal sectional view of the working end 702 corresponding to FIG. 20B wherein the electrical fields EF are confined within the interior lumen 730 to thus cause the explosive vaporization of captured saline. Thus, the second RF mode and the vaporization of captured saline 754 as depicted in FIG. 20B will expel the resected tissue 755 proximally within the tissue extraction channel 730 that extends proximally through the probe to a collection reservoir as described in previous embodiments. In general, a method of the invention includes capturing a tissue volume in a closed distal portion of an interior passageway of an elongate probe and causing a phase transition in a fluid proximate to the captured tissue volume to expand the fluid to apply a proximally directed expelling force to the tissue volume. The time interval for providing a closed window to capture the tissue and for causing the explosive vaporization can range from about 0.01 second to 2 seconds. A negative pressure source also can be coupled to the proximal end of the extraction lumen as described previously. [0087] Now turning to FIG. 22, another variation of inner sleeve 715 ′ is shown. In this embodiment, the leading edge 745 and the trailing edge 745 ′ of electrode edge 725 are provided with different electrical characteristics. In one variation, the leading edge 745 is a highly conductive material suited for plasma ignition as described previously. In this same variation shown in FIG. 22, the trailing edge 745 ′ comprises a different material which is less suited for plasma formation, or entirely not suited for plasma formation. In one example, the trailing edge 745 ′ comprises a resistive material (e.g., a resistive surface coating) wherein RF current ignites plasma about the leading edge 745 but only resistively heats the trailing 745 ′ edge to thus provide enhanced coagulation functionality. Thus, the leading edge 745 cuts and the trailing edge 745 ′ is adapted to coagulate the just-cut tissue. In another variation, the trailing edge 745 ′ can be configured with a capacitive coating which again can be used for enhancing tissue coagulation. In yet another embodiment, the trailing edge 745 ′ can comprise a positive temperature coefficient of resistance (PTCR) material for coagulation functionality and further for preventing tissue sticking. In another variation, the trailing edge 745 ′ can have a dielectric coating that prevents heating altogether so that the leading edge 745 cut tissues and the trailing edge 745 ′ has no electrosurgical functionality. [0088] FIG. 23 illustrates another embodiment of inner sleeve component 718 ′ in which the electrode edge 725 has a leading edge 745 with edge features for causing a variable plasma effect. In this embodiment, the projecting edges 760 of the leading edge 745 electrode will create higher energy density plasma than the scalloped or recessed portions 762 which can result in more efficient tissue cutting. In another embodiment, the electrode surface area of the leading edge 745 and trailing edge 745 ′ can differ, again for optimizing the leading edge 745 for plasma cutting and the trailing edge 745 ′ for coagulation. In another embodiment, the trailing edge 745 ′ can be configured for volumetric removal of tissue by plasma abrasion of the just-cut surface since it wiped across the tissue surface. It has been found that a substantial amount of tissue (by weight) can be removed by this method wherein the tissue is disintegrated and vaporized. In general, the leading edge 745 and trailing edge 745 ′ can be dissimilar with each edge optimized for a different effect on tissue. [0089] FIG. 24 illustrates another aspect of the invention that can be adapted for selective cutting or coagulating of targeted tissue. In this variation, a rotation control mechanism is provided to which can move the inner sleeve 715 to provide the leading edge 745 in an exposed position and further lock the leading edge 745 in such an exposed position. In this locked (non-rotating) position, the physician can activate the RF source and controller to ignite plasma along the exposed leading edge 745 and thereafter the physician can use the working end as a plasma knife to cut tissue. In another variation, the physician can activate the RF source and controller to provide different RF parameters configured to coagulate tissue rather than to cut tissue. In one embodiment, a hand switch or foot switch can upon actuation move and lock the inner sleeve in the position shown in FIG. 24 and thereafter actuate the RF source to deliver energy to tissue. [0090] It should be appreciated that while an RF source is suitable for causing explosive vaporization of the captured fluid volume, any other energy source can be used and falls within the scope of the invention, such as an ultrasound transducer, HIFU, a laser or light energy source, a microwave or a resistive heat source. [0091] In another embodiment, the probe can be configured with a lumen in communication with a remote liquid source to deliver fluid to the interior chamber 240. [0092] Although particular embodiments of the present invention have been described above in detail, it will be understood that this description is merely for purposes of illustration and the above description of the invention is not exhaustive. Specific features of the invention are shown in some drawings and not in others, and this is for convenience only and any feature may be combined with another in accordance with the invention. A number of variations and alternatives will be apparent to one having ordinary skills in the art. Such alternatives and variations are intended to be included within the scope of the claims. Particular features that are presented in dependent claims can be combined and fall within the scope of the invention. The invention also encompasses embodiments as if dependent claims were alternatively written in a multiple dependent claim format with reference to other independent claims.

Summary: Tissue may be cut and extracted from an interior location in a patient&#39;s body using a probe or tool which both effects cutting and causes vaporization of a liquid or other fluid to propel the cut tissue through an extraction lumen of the cutting device. The cutting may be achieved using an electrosurgical electrode assembly, including a first electrode on a cutting member and a second electrode within a cutting probe or tool. Thus, over a first cutting portion, radio frequency current may help cut the tissue and over a second or over transition region, the RF current may initiate vaporization of the fluid or other liquid to propel the tissue from the cutting device.

big_patent

en

Summarize: BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The invention relates to a process for producing porous polymer masterbatch and fiber thereof that have anti-bacterial and odor eliminating functions, and in particular, to a process for producing anti-bacterial and odor eliminating polymer masterbatch and fiber thereof containing organic or inorganic materials. [0003] 2. Description of the Prior Art [0004] As the demand for a modern hygienic life increases, anti-bacterial products have gradually received more welcome from consumers. This trend extends to the textile industry and has resulted into considerable progress in the production of anti-bacterial fibers and clothing and other daily use articles. [0005] Anti-bacterial agent used in the anti-bacterial fiber can be divided generally into two types, namely, an organic anti-bacterial agent, and an inorganic anti-bacterial agent. One of the organic anti-bacterial agents is quaternary ammonium salt. Unfortunately, quaternary ammonium salt has poor heat resistance and can not be used in the process for making plastics or fiber spinning products. [0006] On the other hand, an inorganic anti-bacterial agent is a carrier (for example zeolite) containing metal ions (for example Ag +, Zn 2+, Cu 2+ ), or certain types of nano-scale metal particles (for example nano-scale silver particles), and both are considered as effective particles in the following description. [0007] Silver has a well-accepted anti-bacterial effect. In general, an antibiotic can kill approximately six different types of bacteria, while silver can kill about 600 types of bacteria. In addition, silver is a non-toxic substance, and therefore silver is used extensively and has a long history. Furthermore, through nanotechnology techniques, silver particles become more active and their anti-bacterial function is enhanced, thereby promoting the quality of the home environment and personal hygiene. Aqueous solutions containing silver ions released from both nano-scale silver granules and nanometer silver has remarkable anti-bacterial effect. Under the circumstance of multiple dilutions, nanometer silver still has an inhibition efficiency of 99.99% against Escherichia coli, Staphylococcus aureus, Sarmonella, Pseudomonas aeruginosa and the like. The principal cause of this resides in the biological action that silver has itself. Active silver ions can attract the sulfhydryl group on the enzymatic protein in the bacteria and causes these groups to quickly bind with each other, thereby rendering the enzymes containing sulfhydryl groups to lose activity and hence kill the bacteria. [0008] A traditional process for using silver ion to produce fiber comprises immersing fiber in an organic anti-bacterial agent so as to adhere a carrier or nanometer silver particle on the surface of the fiber. In such traditional processes, effective particles in the anti-bacterial inorganic solvent may be easily washed off, and at the same time, may easily induce an allergic response in the user. Another process for producing anti-bacterial fiber comprises mixing an inorganic anti-bacterial agent and polyester, and then drawing the mixture thus-obtained into fibers containing effective particles. In such a process for making anti-bacterial fiber, most of the anti-bacterial materials are embedded within the fiber, and hence the anti-bacterial and odor eliminating functions are unable to be exhibited. Furthermore, part of the anti-bacterial material exposed on the outside of the fiber might lose its anti-bacterial and odor eliminating functions after washing or dying and finishing due to binding with chlorine, sulfur and the like. [0009] In view of the foregoing, conventional techniques mentioned above still have many disadvantages, poorly designed and needs to be improvement. [0010] The inventor had learned of the various disadvantages and shortcomings derived from such conventional techniques described above, and had thought to improve and innovate, and finally, after studying intensively for many years, has developed a process for producing porous polymer masterbatch and fiber thereof that have anti-bacterial and odor eliminating functions according to the invention. SUMMARY OF THE INVENTION [0011] One object of the invention is to provide a process for producing porous polymer masterbatch and fiber thereof that have anti-bacterial and odor eliminating functions, for the purpose of reducing lost particles in the anti-bacterial inorganic solvent, which tend to be washed off easily, and hence tend to lower the anti-bacterial effect as well as lead to potential unknown effects on the ecological equilibrium of the environment. [0012] Another object of the invention is to provide a process for producing porous polymer masterbatch and fiber thereof that have anti-bacterial and odor eliminating functions, characterized in that the inventive process can improve the previous conventional process for making anti-bacterial fiber; where in the conventional process, an inorganic anti-bacterial agent is mixed in a polymer and then the resulted mixture is drawn to form fiber containing fine nanometer particles, and in such conventional process for making anti-bacterial fiber, the nanometer particles are difficult to be dispersed homogeneously in the fiber, and further, most of the nanometer particles are embedded within the fiber, so that its anti-bacterial effect can not function effectively. [0013] Still another object of the invention is to provide a process for producing porous polymer masterbatch and fiber thereof that have anti-bacterial and odor eliminating functions, characterized in that wide and diverse materials can be used, and has a wide spectrum of anti-bacterial and anti-fungal effects. [0014] The process for producing porous polymer masterbatch and fiber thereof that have anti-bacterial and odor eliminating functions comprises: step 1, grinding kieselguhr or active carbon into micro-particles; step 2, immersing said micro-particles obtained in step 1 in an organic Chinese herbal medicine and inorganic anti-bacterial minerals and polyvinyl alcohol with constant stirring; step 3, air-drying micro-particles thus-obtained in step 2, and dry-grinding further the dried micro-particles into finer particles suitable for drawing and to be dispersed homogeneously in a solution; step 4, carrying out a esterification reaction for binding monomer to form an anti-bacterial polyester masterbatch; and/or step 5, producing the polyester masterbatch formed in step 4 into anti-bacterial polyester fiber by cold grain spin-drawing technique. BRIEF DESCRIPTION OF THE DRAWINGS [0015] These features and advantages of the present invention will be fully understood and appreciated from the following detailed description of the accompanying drawings, wherein: [0016] FIG. 1 shows the flow chart for carrying out the process according to the invention; [0017] FIG. 2 is a schematic view of porous micro-particles according to the invention; [0018] FIG. 3 is a schematic view of finer micro-particles obtained after dry-grinding according to the invention; and [0019] FIG. 4 is a schematic view of anti-bacterial polyester fibers according to the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT [0020] For understanding further the objects, characteristics and effects of the invention, the following non-limiting examples will be illustrated in conjunction with the accompanied drawings below. [0021] Referring to FIG. 1, the process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions provided according to the invention comprises following steps: [0022] step 1 : grinding kieselguhr or active carbon into micro-particles of 1˜1000 micron ( 111 ); Referring to FIG. 2, a schematic view of kieselguhr or active carbon micro-particles. In the step 1, kieselguhr or active carbon is ground at first into micro-particles 1. In the case of active carbon, the principal component of active carbon is carbon, combined with minor amount of hydrogen, oxygen, nitrogen, sulfur and the like, and is a black porous material 2 with complex surface. It has a six ring structure formed from carbon and its shape may range from a cylindrical coarse granule to a fine powder particle, and hence it has two type of morphology of granule and powder. The granule has generally a diameter of 1˜6 mm, and its length is 0.7˜4 times of its diameter. Alternatively, it may be present as a granule of irregular shape with 6˜120 particle mesh. The active carbon is odorless, and tasteless, and is insoluble in water and organic solvent. Active carbon has a packing density of about 0.3˜0.6 g/ml, a volume of the very large micro-pore of about 0.6˜0.8 ml/g, and a specific surface of about 500˜1,500 m 2 /g, and therefore, exhibits a very strong adhesive force to an organic macromolecular material; In addition, kieselguhr can be used in the process of the invention. Kieselguhr is mined from fossil lake beds. Such kieselguhr is formed from the deposition of large amounts of dead micro-diatoms on the ancient lake or sea bed. Diatom is classified into two morphologies of salt water diatom and fresh water diatom. Accordingly, the thus-mined kieselguhr is classified also into two types; salt water kieselguhr and fresh water kieselguhr. After mining, it can be washed, processed and ground into micro-powder of various specific sizes; [0025] step 2 : impregnating micro-particles obtained in step 1 in an organic Chinese herbal medicine and a water-soluble cementing agent, adding inorganic anti-bacterial minerals of 1˜100 nm in size, and then stirring continuously for more than 24 hours, thereby enabling the organic Chinese herbal medicine and inorganic anti-bacterial minerals to be impregnated, bound and adhered sufficiently in the pores of the micro-particle ( 112 ); Referring to Table 1, each of the inorganic anti-bacterial minerals mentioned above has its own special effect. Inorganic anti-bacterial minerals used in the process of the invention may be selected from the group consisting of realgar, calamine, melanterite, talc, alum, sulfur, borax, nanometer silver (Ag), nanometer zinc (Zn), nanometer copper (Cu), titanium dioxide (TiO 2 ) and any combination thereof; [0000] TABLE 1 The pharmacological effect, function and chemical component of inorganic anti-bacterial minerals. Item Name Pharmacological effect 1. Realgar Detoxification, removing moisture, killing insects, anti-bacterial 2. Calamine Absorbing moisture, stopping itch, controlling sores, antiseptic, anti-bacterial 3. Melanterite Treating eczema, killing insects, anti-bacterial 4. Talc Treating eczema, wet sore, scabies, anti-bacterial 5. Alum Detoxification, killing insects, drying moisture, stopping itch, wide spectrum anti-bacterial 6. Sulfur Killing insects, stopping itch, scabies, eczema, killing insects, anti-bacterial 7. Borax Detumescence, antiseptic, anti-bacterial, treating scabies and itch 8. Nanometer Anti-bacterial, anti-fungal silver (Ag) 9. Nanometer Anti-bacterial, anti-fungal zinc (Zn) 10. Nanometer Anti-bacterial, anti-fungal copper (Cu) 11. Titanium Anti-bacterial, anti-fungal dioxide (TiO 2 ) The water-soluble cementing agent 3 may be selected from polyvinyl alcohol (PVA) or the like. PVA is an extensively used water-soluble macromolecular polymer, with a property between those of plastic and rubber. Since PVA possesses a strong bonding property, flexibility, smoothness of the surface texture, oil resistance, solvent resistance, protective gel property, gas insulating property, wear resistance and after special treatment, water resistance, it is used very often in raw materials for fiber; The binding property of PVA is utilized to adhere the extract essence fluid of herbal plants on the surface of the outer and inner holes of micro-particles so as to increase the amount and surface area of the organic Chinese herbal medicine and inorganic anti-bacterial minerals thus-adhered. The herbal plants useful in the process according to the invention may be selected from the group consisting of Radix et RhizomanNotopterygii, Black false hellebore, Hibiscus syriacus skin, Cinnamomum cassia Presl, camphor, Cnidium monnieri (L) Cuss, Hydnocarpus anthelmintica Pier., rosin and any combination thereof. Each of the herbal plants mentioned above has its own specific effect as shown in Table 2; [0000] TABLE 2 Pharmacological effect, functions and chemical components of Chinese herbs Pharmacological Item Name effect Function Chemical components 1. Radix et Rhizoma Anti-bacterial, notopterygii anti-fungal 2. Black false hellebore Anti-bacterial, anti-fungal, killing insects 3. Hibiscus syriacus Killing insects, skin stopping itch, anti-bacterial 4. Cinnamomum cassia Anti-bacterial, Sedation, Cinnamic aldehyde, Presl anti-bacterial Analgesia, Cinnamic acid, anti-tinea, Allaying fever Cinnamyl acetate, anti-fungal Phenylpropyl acetate 5. Camphor Removing moisture, killing insects, anti-bacterial, anti-fungal 6. Cnidium monnieri Anti-fungal, Removing L-Piuene, (L) Cuss anti-Gram negative rheumatism, L-Camphene, bacteria, anti-mold drying Bornyl isovalerate, Anti-Ringworm moisture, Isoborneol, Edultim, fungus killing insects, Cnidimine, stopping itch Xanthotoxin 7. Hydnocarpus Anti-bacterial, Detoxification, Chaulmoogric Acid, anthelmintica Pier. treating tinea killing insects, Hydnocarpic Acid, manus and tinea removing tinea Gorlic Acid pedis cruris 8. Rosin Anti-bacterial, anti-mold, treating scabies, wet itch The organic Chinese herbal medicine and inorganic anti-bacterial minerals mentioned in step 2 may be used in combination with one another to achieve the desired pharmacological effect and function; [0030] step 3 : air drying the micro-particle impregnated in step 2, and dry grinding them further into finer micro-particle of 1˜50 micron in size suitable for drawing, and dispersing them homogeneously in a solution ( 113 ); Referring to FIG. 2, the surface and the outside and inside of the holes in the thus-impregnated micro-particles had been adhered with extract essence fluid of the herbal plants. Thereafter, the impregnated micro-particles thus air dried were dry ground further into finer micro-particles. The surface 6 and the outside and inside of holes 4 in the thus-obtained finer micro-particles 7 had been adhered with the organic Chinese herbal medicine and inorganic anti-bacterial minerals 5. The air-dried finer micro-particle 7 had a size of 1˜50 micron and became a finer micro-particle suitable for drawing as well as could be dispersed homogeneously in a solution. The solution mentioned in the step 3 of the inventive process may be ethylene glycol solution. Ethylene glycol is a catalyst used in the condensation polymerization of polyester, and has the following advantages: 1. It has greater solubility and better dispersability in ethylene glycol solution; 2. It has good activity, and can enhance productivity of the apparatus; 3. This catalyst itself will not introduce new contaminants, can increase the intrinsic mass and improve post-processing spinning ability; 4. It can improve hues and heat stability of the slice. [0033] step 4 : binding the finer micro-particle dispersed homogeneously in the solution to a monomer through esterification reaction to form anti-bacterial polyester masterbatch, wherein such finer micro-particle in each of the polyester masterbatch comprises about 10-25% of the total weight of each masterbatch ( 114 ); [0034] step 5 : Using low temperature batch processing chip spinning technique to produce the anti-bacterial polyester masterbatch to produce anti-bacterial polyester fiber 8, wherein the ratio of the anti-bacterial polyester masterbatch added is about 5-10% ( 115 ). [0035] In summary, the process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions provided by the invention comprises following steps: [0036] step 1 : grinding kieselguhr or active carbon into micro-particles of 1˜1000 micron in size ( 111 ); [0037] step 2 : impregnating micro-particles obtained in step 1 with organic Chinese herbal medicine and water-soluble cementing agent and inorganic anti-bacterial minerals of 1˜100 micron in size, and stirring continuously for more than 24 hours, thereby enabling sufficient penetration, binding and adsorption of the organic Chinese herbal medicine and inorganic anti-bacterial minerals in pores of micro-particles ( 112 ); [0038] step 3 : air drying micro-particles impregnated in step 2, and then dry grinding said dried micro-particles into finer micro-particle of 1˜50 micron in size suitable for drawing and dispersing homogeneously in a solution ( 113 ); [0039] step 4 : binding said finer micro-particles dispersed homogeneously in a solution with monomer through esterification reaction to form said anti-bacterial polyester masterbatch, wherein such finer micro-particle in each of the polyester masterbatch comprises about 10-25% of the total weight of each masterbatch ( 114 ); [0040] step 5 : producing anti-bacterial polyester fiber from said anti-bacterial polyester masterbatch by low temperature batch processing chip spinning technique, wherein the ratio of the anti-bacterial polyester masterbatch added is about 5-10% ( 115 ). [0041] Accordingly, the process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions provided by the invention has following advantages over other conventional techniques: [0042] 1. The inventive process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions can reduce the lost amount of particles originally present in the anti-bacterial inorganic solvent, wherein said particles is susceptible to be washed off, and thus, may lower its anti-bacterial effect as well as may cause an unexpected influence on the ecological equilibrium of the environment. [0043] 2. The inventive process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions can enable these odor eliminating anti-bacterial materials to penetrate and adhere into tremendous micro-pores of these porous particles, and further, since they can bind in these micro-pores by means of water-soluble cementing agent, thereby these materials tend not to lose and hence can achieve the purpose of increasing its anti-bacterial and odor eliminating functions. [0044] 3. The inventive process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions can be extended to make various related products having anti-bacterial and odor eliminating functions, such as shoes, bags, socks, clothes. [0045] 4. The inventive process for producing porous polymer masterbatch and fiber thereof having anti-bacterial and odor eliminating functions can adopt wide and diverse materials, and hence can retain wide spectrum anti-bacterial and anti-fungal effects, without loses its anti-bacterial function during dying and finishing process due to bind with chlorine and sulfur like, for example, using nanometer silver alone. [0046] Many changes and modifications in the above-described embodiment of the invention can, of course, be carried out without departing from the scope thereof. Accordingly, to promote the progress in science and useful arts, the invention is disclosed and is intended to be limited only by the scope of the appended claims.

Summary: A process for producing porous polymer masterbatch having anti-bacterial and odor eliminating functions, wherein said fiber contains materials such as porous natural mineral kieselguhr or active carbon, can absorb and eliminate odor such as stink of sweat and the like discharged from the human body, has functions of sterilization, anti-bacterial, anti-mold, and the like. Pores of the natural mineral kieselguhr or active carbon contain organic Chinese herbal medicine and inorganic anti-bacterial minerals, wherein all of such organic Chinese herbal medicine and inorganic anti-bacterial minerals have functions of anti-bacterial, anti-fungal and the like, can eliminate effectively odor or reduce substantially stink, and can be applied extensively on various fabrics, clothes and ornaments or other goods.

big_patent

en

Write a title and summarize: SECTION 1. SMALL BUSINESS AND AGRICULTURAL PRODUCER ENERGY EMERGENCY DISASTER LOAN PROGRAM. (a) Small Business Producer Energy Emergency Disaster Loan Program.--Section 7(b) of the Small Business Act (15 U.S.C. 636(b)) is amended by adding at the end the following new paragraph: ``(4) Energy emergency disaster loans.-- ``(A) Authority.--The Administrator may make or guarantee a loan to a small business concern that the Administrator determines has suffered or is likely to suffer substantial economic injury as a result of a significant increase in the price of heating oil, natural gas, gasoline, transportation fuel, propane, or kerosene. ``(B) Interest rates.--Any loan or guarantee extended pursuant to this paragraph shall be made at the same interest rate as an economic injury loan made or guaranteed under paragraph (2). ``(C) Limitation.-- ``(i) In general.--No loan may be made or guaranteed under this paragraph if the total amount outstanding and committed to the borrower under this subsection would exceed $1,500,000. ``(ii) Exception.--The Administrator may waive the limitation under clause (i) for a borrower if the Administrator determines that the borrower constitutes a major source of employment in its surrounding area. ``(D) Declarations of disaster.--No assistance shall be available under this paragraph unless-- ``(i) the President or the Administrator has made a declaration of a disaster area by reason of a significant increase in the price of heating oil, natural gas, gasoline, transportation fuel, propane, or kerosene; or ``(ii) the Governor of a State in which a significant increase in the price of heating oil, natural gas, gasoline, transportation fuel, propane, or kerosene has occurred certifies to the Administrator that small business concerns have suffered economic injury as a result of such increase and are in need of financial assistance that is not otherwise available on reasonable terms in that State. ``(E) Conversion to renewable or alternative energy sources.--Notwithstanding any other provision of law, a small business concern receiving a loan under this paragraph may use the loan to convert from the use of heating oil, natural gas, gasoline, propane, or kerosene to a renewable or alternative energy source, including agricultural and municipal solid waste, geothermal energy, cogeneration, solar energy, wind energy, or fuel cells. ``(F) Definitions.--In this paragraph, the following definitions apply: ``(i) The term `base price index' means the moving average of the closing unit price on the New York Mercantile Exchange for heating oil, natural gas, gasoline, transportation fuel, or propane for the 10 days, in each of the most recent 2 preceding years, which correspond to the trading days described in clause (ii). ``(ii) The term `current price index' means the moving average of the closing unit price on the New York Mercantile Exchange, for the 10 most recent trading days, for contracts to purchase heating oil, natural gas, gasoline, transportation fuel, or propane during the subsequent calendar month, commonly known as the `front month'. ``(iii) The term `significant increase' means-- ``(I) with respect to the price of heating oil, natural gas, gasoline, transportation fuel, or propane, an increase of the current price index over the base price index by not less than 40 percent; and ``(II) with respect to the price of kerosene, any increase which the Administrator, in consultation with the Secretary of Energy, determines to be significant.''. (b) Effective Date.--The amendment made by this Act shall apply with respect to economic injury suffered on or after the date of the enactment of this Act.

Title: To amend the Small Business Act to establish an energy emergency disaster loan program Summary: Amends the Small Business Act to authorize the Administrator of the Small Business Administration (SBA) to make or guarantee a loan to a small business that has suffered, or is likely to suffer, substantial economic injury as a result of a significant increase in the price of heating oil, natural gas, gasoline, transportation fuel, propane, or kerosene. Prohibits such a loan or guarantee if the total amount outstanding and committed to the borrower would exceed $1.5 million (with an exception). Prohibits any such assistance unless there has been a declaration of a disaster in the area or the governor of the state involved has certified that small businesses have suffered economic injury as a result of the price increases. Allows a small business to use assistance funds to convert to a renewable or alternative energy source.

billsum

en

Write a title and summarize: Cryptococcus neoformans is a heterothallic fungal pathogen of humans and animals. Although the fungus grows primarily as a yeast, hyphae are produced during the sexual phase and during a process called monokaryotic fruiting, which is also believed to involve sexual reproduction, but between cells of the same mating type. Here we report a novel monokaryotic fruiting mechanism that is dependent on the cell cycle and occurs in haploid cells in the absence of sexual reproduction. Cells grown at 37°C were found to rapidly produce hyphae (∼4 hrs) and at high frequency (∼40% of the population) after inoculation onto hyphae-inducing agar. Microscopic examination of the 37°C seed culture revealed a mixture of normal-sized and enlarged cells. Micromanipulation of single cells demonstrated that only enlarged cells were able to produce hyphae and genetic analysis confirmed that hyphae did not arise from α-α mating or endoduplication. Cell cycle analysis revealed that cells grown at 37°C had an increased population of cells in G2 arrest, with the proportion correlated with the frequency of monokaryotic fruiting. Cell sorting experiments demonstrated that enlarged cells were only found in the G2-arrested population and only this population contained cells able to produce hyphae. Treatment of cells at low temperature with the G2 cell cycle arrest agent, nocodazole, induced hyphal growth, confirming the role of the cell cycle in this process. Taken together, these results reveal a mating-independent mechanism for monokaryotic fruiting, which is dependent on the cell cycle for induction of hyphal competency. Cryptococcus neoformans is a basidiomycetous fungal pathogen of humans and animals that typically causes opportunistic infections in patients with cellular immune defects [1]. Infection initiates in the lungs and frequently disseminates to the brain where it manifests as a fatal meningoencephalitis if untreated. AIDS patients are at increased risk for infection, though infection rates have decreased significantly with better AIDS management [2]. However, in spite of the reduction in AIDS-related cases, cryptococcosis remains a frequent life-threatening opportunistic mycosis for these patients in underdeveloped countries, and is a recently emergent disease in the United States Pacific Northwest [3] for as yet, unexplained reasons. Naturally occurring strains of C. neoformans are heterothallic with two mating types, MATa and MATα, with both mating types being pathogenic, although most clinical isolates are MATα [4]. Although the taxonomy has been changing, four serotypes have been described (A, B, C, D) with serotypes A and D often referred to as C. neoformans variety grubii and variety neoformans respectively, and serotypes B and C being collectively referred to as C. neoformans variety gattii, or more recently, C. gattii [5]. For all serotypes, throughout the course of infection and under normal culture conditions, the fungus grows as an encapsulated yeast. Under appropriate in vitro conditions, however, the fungus can produce two kinds of hyphae; dikaryotic hyphae during MATa×MATα sexual reproduction and monokaryotic hyphae (from individual MATa or MATα strains) during monokaryotic fruiting [6]. Basidiospores can be produced from both hyphal types. Environmental factors required for sexual reproduction and monokaryotic fruiting are similar and include culture under low temperature (25°C), low moisture, and nutrient limitation [6]. Many genes, including homologs of the Saccharomyces cerevisiae pheromone response pathway, are required to produce both types of hyphae [7], [8], [9], [10], [11]. There are, however, distinct differences between the two hyphal types. Structurally, dikaryotic hyphae have fused clamp connections and a pair of nuclei (one MATa and one MATα) per hyphal compartment while monokaryotic hyphae have unfused clamp connections and a single nucleus per hyphal compartment. Sexual reproduction in C. neoformans has been characterized in detail and largely follows the pheromone response paradigm that has been developed from decades of S. cerevisiae research [12]. Less clear is the mechanism by which monokaryotic fruiting occurs. Recent studies have concluded that monokaryotic fruiting in C. neoformans variety neoformans can result from α-α mating [13] and may be an important part of the natural life cycle of this fungus [14], with possible implications for human disease [15]. We recently reported that high temperature seed culture conditions could induce very robust monokaryotic fruiting in C. neoformans variety neoformans [11] and assumed these growth conditions enhanced α-α cell fusion. However, this assumption proved to be false. Instead, we found that cells arrested in the G2 stage of the cell cycle were competent to undergo monokaryotic fruiting at high frequency, in the absence of α-α cell fusion. Importantly, this mechanism proceeds through enlarged cells, a morphological phenotype that has been increasingly observed in vivo and is hypothesized to serve as a strategy for avoiding host defenses [16], [17], [18]. These results demonstrate that C. neoformans has evolved a number of different mechanisms for modifying cellular morphology to suit its specific environment, with some of these mechanisms contributing to the success of this fungus as a pathogen. Previous studies of monokaryotic fruiting typically were performed by patching cells from a seed culture onto filament agar and then screening for a hyphal fringe around the periphery [6], [11], [19]. Our recent observation of the role of temperature in this process [11] led us to test whether or not high temperature increases the intensity of hyphae production after inoculation onto filament agar, or increases the number of cells that produce hyphae. A suspension of cells from a 24 h, 37°C seed culture was spread onto filament agar to observe monokaryotic fruiting in individual cells, which would reveal whether or not all cells from the seed culture were capable of undergoing monokaryotic fruiting. Figure 1A demonstrates that only part of the population grown at 37°C was able to undergo monokaryotic fruiting and that hyphae began to appear as early as 4 hours after plating onto filament agar (Fig. 1B). This phenomenon was not a strain artifact nor was it restricted to a single serotype as all four serotypes were found to be able to produce hyphae under the above inducing conditions (Fig. 1C–F). These results suggest that only a specific type of cell is capable of undergoing monokaryotic fruiting, and that this capability requires seed culture conditions that enable these cells to become competent for hyphal production. Lin et al., have demonstrated that same-sex mating is one way in which monokaryotic fruiting can occur [13]. However, because the previous experiment showed that individual cells were still capable of monokaryotic fruiting in spite of being well separated from potential mating partners on spread plates, we hypothesized that either same-sex mating occurs during the seed culture growth period, or that another mechanism, which does not involve same-sex mating, could also lead to monokaryotic fruiting. To investigate these two possibilities, we first screened for evidence of α-α mating during monokaryotic fruiting using the α-α cell fusion assay to test different combinations of complementing MATα auxotrophs that would be predicted to yield prototrophic colonies on unsupplemented MIN agar. No fusants were observed from MATα×MATα crosses plated onto MIN agar after growth as a seed culture on YPD at 37°C for 24 h. However, assisted α-α matings showed that each of these strains was capable of undergoing α-α fusion, ruling out an α-α cell fusion defect (Fig. 2A). The most likely explanation for these results was that because α-α fusion is a rare event [13], our conditions, although not detecting an α-α fusion, were still not excluding this possible mechanism. To exclude a cryptic fusion event as an explanation for our observations, we utilized a cpk1Δ mutant to further test whether or not α-α fusion was required for temperature-induced monokaryotic fruiting. Cpk1p, the MAP kinase in the C. neoformans pheromone response pathway, is required for α-a fusion during sexual reproduction as well as α-α fusion during monokaryotic fruiting [9], [13]. Therefore, if α-α fusion was the mechanism of monokaryotic fruiting in our system, the cpk1Δ mutant should not produce hyphae after plating onto filament agar because it could not fuse with the complementing strain. Our results showed that neither assisted nor unassisted α-α mating reactions (WSA-2126×WSA-591×WSA-65 or WSA-2126×WSA-591) with the cpk1Δ mutant (WSA-2126) showed evidence of α-α cell fusion (Fig. 2B). However, when WSA-2126 was tested for monokaryotic fruiting ability after high temperature seed culture, this strain fruited normally (Fig. 2B). These results confirmed that monokaryotic fruiting could occur independently of α-α fusion. The production of enlarged cells in C. neoformans has been reported to occur when cells are exposed to opposite mating type cells in standard mating type mixes [11], and in confrontation assays, which are performed by streaking cells of opposite mating type in close proximity to each other [20]. Clinical studies have also found this cell type in vivo [16], [18], [21], [22]. Because only certain cells produced hyphae during the quantitative monokaryotic fruiting assay, we decided to determine if there were developmental differences among cells after high temperature seed culture. Microscopic observation of wet mounts prepared from cells scraped off of filament agar after growing as a seed culture at 37°C showed that filaments always originated from enlarged cells (Fig. 3A). When cells from 30°C and 37°C seed cultures were screened for enlarged cells, we only observed enlarged cells from the 37°C seed culture, although this incubation temperature produced both enlarged and smaller, normal-sized cells (Fig. 3B and 3C). To confirm that the enlarged cells were responsible for the production of hyphae during monokaryotic fruiting, large and small cells from a 37°C seed culture were micromanipulated onto filament agar and then monitored for hyphae production. Analysis of hyphae production from each single cell revealed that small cells only grew into yeast cells while the large cells grew as both yeast and hyphae (Fig. 3D). These results demonstrated that only a subset of the population grown at high temperature, which can be distinguished by the larger cell size, becomes competent to produce hyphae. The need for enlarged cells prior to the initiation of monokaryotic fruiting suggested that the mechanism for production of this phenotype possibly involved changes in cell ploidy since yeast ploidy has been noted to be associated with cell size [23]. In C. neoformans, monokaryotic fruiting has been shown to result in ploidy changes of yeast cells produced specifically from the hyphal filaments [13]. These observations suggested to us that monokaryotic fruiting may occur through ploidy changes, which are manifested as enlarged cells that arise from an endoduplication event, as has been previously suggested [24]. Cell cycle analysis of seed cultures grown at different temperatures (25°C, 30°C, 35°C, 37°C and 40°C) all revealed only 1n and 2n DNA content peaks, with no evidence of a 4n peak (Fig. 4A). This result showed that there was no ploidy change after high temperature (35°C, 37°C, 40°C) seed culture growth, demonstrating that the enlarged cells, which were responsible for monokaryotic fruiting, were haploid. Additionally, we found that as seed culture temperature increased from 25°C to 40°C, the percentage of cells in G1 decreased, the percentage of cells in G2 increased, and the percentage of cells in S phase was similar until dropping almost to 0 at 37°C (Fig. 4B). DAPI staining confirmed the relationship between cell size and G2 arrest as the staining patterns of large and small cells differed (Fig. 4C). The smaller cells displayed a compact nuclear staining pattern while the larger cells displayed a larger, diffuse staining pattern, which has been observed in other G2-arrested fungi [25], [26]. These results demonstrated that the effect of increasing temperature on monokaryotic fruiting involved the cell cycle, specifically G2 arrest. In spite of the FACS results showing that hyphal-competent cells were haploid, we could not rule out a change in ploidy just before, or during growth in the hyphal phase. Unfortunately ploidy determination by FACS analysis of hyphae is physically restricted by the filamentous characteristics of the cells. However, hyphal ploidy can be determined using a blastospore assay [13]. The results of this assay revealed that 76 out of 78 blastospores (97%) were haploid, demonstrating that the hyphae produced during monokaryotic fruiting were haploid, which again excluded α-α mating or endoduplication as monokaryotic fruiting mechanisms in our system (Fig. 5A). As a further control, the two diploid blastospores were sub cultured repeatedly for an additional two weeks and then retested by FACS, which revealed that they remained diploid, ruling out the possibility that haploid blastospores could be segregation products of diploid blastospores. While a late endoduplication event could explain the two diploid spores, this possibility is unlikely since the 78 spores were picked from 78 independent hyphae. However based on the recovery of two diploid blastospores in our assay, and the rarity of basidiospore production during monokaryotic fruiting, we hypothesized that there could be two hyphal types produced during monokaryotic fruiting, which could be distinguished by ploidy. One type could be vegetative in nature and not undergo basidiosporogenesis (haploid) and a second type could be generated that ultimately produced basidiospores (diploid). To address these two possibilities, 37°C seed culture cells were spread onto filament agar and the resultant colonies screened for basidiospore chains. The blastospore assay was performed on blastospores recovered from hyphae with and without basidiospores. Cell cycle analysis again showed that all of the blastospores were haploid, regardless of whether or not the hypha produced basidiospores (Fig. 5B), which was consistent with our previous observations that excluded α-α mating or endoduplication as the mechanisms of monokaryotic fruiting. Together, these results demonstrate that endoduplication is not required for monokaryotic fruiting as these hyphae are produced from haploid cells and remain haploid, in spite of being able to occasionally generate diploid yeast cells. To determine whether only G2-arrested cells undergo monokaryotic fruiting, we sorted G1 and G2 phase, 37°C seed culture cells according to DNA content. Microscopic observation of sorted G1 and G2 cells revealed that G2 phase cells were much larger than G1 phase cells (Fig. 6). We next sorted live 37°C seed culture cells according to cell size and performed cell cycle analysis on the two populations. The results indicated that the smaller cells had a single DNA content peak at the 1n position while the enlarged cells had a single DNA content peak at the 2n position, which lead us to conclude that the small cells were G1 phase cells and the large cells were cells in G2 arrest (Fig. 7). The two populations were then assayed for monokaryotic fruiting ability, which revealed that monokaryotic fruiting was a property only of the G2 fraction (Fig. 7). As a final confirmation that monokaryotic fruiting requires G2 arrest, cells were treated with nocodazole, a G2/M arrest agent that inhibits and disassembles microtubules [27]. Cells were grown as seed cultures at 30°C (the non-permissive seed culture temperature), then assayed for monokaryotic fruiting ability. The experiment showed that cells treated with nocodazole became competent to produce hyphae on filament agar in a dose-dependent manner even though they were grown as a seed culture under conditions that did not normally lead to monokaryotic fruiting, whereas untreated cells only grew as yeasts (Fig. 8). Taken together, these results demonstrate that G2-arrested cells can serve as a starting point for cells that undergo monokaryotic fruiting. In this study we have identified a novel mechanism in C. neoformans that leads to the production of hyphae, with or without basidiospores, by haploid cells (monokaryotic fruiting). This mechanism appears to be dependent on the cell cycle and initiates from cells in G2 arrest. It occurs in the absence of α-α mating and/or endoduplication, thereby demonstrating that monokaryotic fruiting can occur asexually. Previous studies have shown that sexual reproduction can occur between cells of the same mating type, resulting in monokaryotic fruiting [13], and that this phenomenon occurs in nature [24]. Under the specific conditions of this study, notably a 37°C seed culture temperature, we saw no evidence of an α-α cell fusion event, nor did we find evidence of endoduplication within the hyphae even though we screened hyphae that had produced basidiospores. During C. neoformans basidiosporogenesis, sexual reproduction results in meiosis in the basidium followed by successive mitotic divisions that yield the nuclei, which ultimately are inserted into spores as they form on the basidial surface [20]. Lin et al. observed that when fruiting was derived from an α-α fusant, sporogenesis was robust with spore chains that were long and phenotypically similar to α-a mating during sexual reproduction [13]. This process was found to be impaired in dmc1 mutants, which are meiotic mutants that still produce spores, but at a much lower frequency than sexually produced spores, and with truncated spore chains that sometimes occur as dyads (two rather than four chains) [13]. The phenotype of basidiospores produced in the dmc1 strains was strikingly similar to what we observed in this study and what was previously reported [6]. These observations may suggest that basidiosporogenesis can occur mitotically without meiosis, although we cannot exclude a duplication event in the basidium immediately followed by meiosis and sporogenesis. We did, however, test a dmc1 mutant and found that it was able to undergo monokaryotic fruiting under our conditions (data not shown). Because our study was done in the same strain background as the study by Lin et al., we reviewed the conditions of both experiments and found some differences that may explain the contrasting differences in ploidy. Our study used a high temperature seed culture condition, which results in rapid hyphae production upon filament agar plating. The seed culture conditions in Lin' s study were not clear, however, their plating medium was V8 agar, which is normally used for mating C. neoformans, and their incubation period was for a period of weeks, whereas we screened at 24 hrs and observed hyphae in as little as four hours, although cells also produced hyphae on V8 agar under our conditions. Both filament agar and V8 agar have high agar contents; however, V8 agar is an undefined medium with V8 juice as the basal ingredient. Filament agar, on the other hand, is a low-glucose, defined medium with Yeast Nitrogen Base without amino acids and without ammonium sulfate as the source of vitamins and cofactors. Although both media are starvation media, they are substantially different in composition, which may be one explanation for the differences in hyphal types that we observed. The seed culture conditions, or more precisely, the cell cycle stage may be another explanation. Our initial investigation of the relationship between the cell cycle and monokaryotic fruiting focused on detecting what we presumed would be a transition to diploidy in yeast cells at some point during seed culture growth, which we believed would coincide with the appearance of enlarged cells in the seed culture and the association of this cell type with the hyphal progenitor. The high frequency of fruiting and enlarged cells in the seed culture suggested that detection of the diploidization event would be unambiguous. However, the data showed that instead of an α-α fusion or endoduplication event, which would result in a diploid cell, the actual mechanism that resulted in hyphae production was G2 arrest. The reason for the requirement of G2 arrest to induce hyphae is not clear, and while G2 arrest is required for monokaryotic fruiting, not all arrested cells produced hyphae. We suspect that the subpopulation of non-hyphal, G2-arrested cells consisted of cells that escaped G2, and then proceeded to bud rather than differentiate into a hypha. These two outcomes resemble the decision point that a pheromone-exposed, G2-arrested Ustilago maydis cell faces with regard to which of the two developmental paths it will follow (conjugation tube formation or budding) [26]. How the generation of occasional diploid blastospores occurs is also not clear. Given that C. neoformans hyphae produce typical basidiomycete-like clamp connections, the incomplete fusion of these structures in monokaryotic hyphae combined with an aberrant segregation event during the budding of blastospores off of the hyphal compartment may yield the diploid cells that we observed at low frequency. Previous studies of the C. neoformans cell cycle have identified a number of stressors that cause G2 arrest, including oxygen depletion [28], stationary growth phase [29], and temperature [30]. Under our experimental conditions, hyphal competency occurs prior to transfer to hyphal inducing conditions (starvation on filament agar) and not during growth on filament agar. Therefore, stationary growth phase is not a factor nor is oxygen depletion since cultures were grown on the agar surface, and only for 24 hrs. Consequently, growth temperature seems to be responsible for inducing competency. Under our conditions, the 37°C incubation temperature differs from the original incubation temperature (30°C) for monokaryotic fruiting [6], which suggests that temperature is an important variable. In fact, while we did not see the temperature effect on all strains of C. neoformans, we were able to induce hyphae in all four serotypes. Interestingly, the reports of enlarged cells in vivo [16], [18], [21], [22] reflect growth at elevated temperature in the mammalian body. Other pathways have also been shown to influence C. neoformans cell size in vitro, including cAMP, RAS, and PKA [16], [31], [32], suggesting the possibility of conserved stimuli that may regulate these pathways. Presently, the STE12α signal transduction pathway seems to be the major or sole regulator of monokaryotic fruiting as this gene is required for monokaryotic fruiting regardless of inducing conditions. Fruiting still occurred normally in cpkΔ1 (pheromone response pathway), and cacΔ1 (cAMP pathway) mutants, ruling out these pathways as regulators of temperature-induced monokaryotic fruiting. Other pathways, such as the calcineurin signal transduction pathway cannot be ruled out, but are more complicated to test since some mutants in this pathway do not grow at high temperature [33]. A key characteristic of enlarged cells in vivo appears to be polyploidy, which has been hypothesized to arise when the M phase of the cell cycle is skipped [16], [18]. This enlarged cell phenotype appears to be a potential virulence factor as they are poorly phagocytized, if at all [18]. Perhaps increasing cell size evolved as a physical defense mechanism against predatory grazers, which in turn, protects cells from being phagocytized in vivo via the same mechanisms. It appears that the enlarged cell phenotype can be produced by multiple mechanisms: cell cycle arrest, a-α or α-α cell fusion, and endoduplication, each of which may have a different purpose. The developmental options available after cell cycle arrest may have been selected for in C. neoformans to enhance survival in its specific environmental niche while inadvertently creating an important human fungal pathogen. What remains to be determined is how high temperature generation of the large cell and monokaryotic fruiting phenotypes was incorporated into the evolution of C. neoformans. With the exception of C. neoformans, virtually all members of this genus grow poorly or not at all at mammalian ambient temperature. In contrast, all of the major human fungal pathogens grow at 37°C and virtually all of them have a hyphal phenotype. Perhaps the association of many basidiomycetes with rotting wood or decaying vegetation in general led to high temperature exposure and subsequent genetic selection during self-heating, compost-like conditions caused by microbial metabolism of organic matter. Once nutrients were consumed, hyphal extension towards additional nutrients and/or sporulation in the absence of nutrients could have completed the evolution of C. neoformans into a pathogen via development of a mechanism of infectious particle (basidiospores) dispersion combined with the ability to grow at elevated temperatures. Selection for the molecular linkage of pathways controlling cell cycle, nutrient sensing, and ultimately, differentiation, could have been the outcome of this lifestyle and allowed the fungus to coordinately regulate these pathways, thus enabling it to effectively exist as a saprophyte or pathogen. YPD agar, MIN agar, V8 agar, and filament agar were prepared as described previously [6], [7], [11] with or without amino acids or nucleic acid supplements as required. JEC-21 is a wild type, MATα isolate that was used in the initial characterization of monokaryotic fruiting in C. neoformans [34]. WSA-79 is a serotype D clinical isolate from Maryland, WSA-522 is a serotype A clinical isolate from Thailand, WSA-533 is a serotype B environmental isolate from Australia, and WSA-2507 is a serotype C clinical isolate from Maryland. Additional strains, all of which were derived from the original JEC-21 - JEC-20 congenic pair [34], consisted of the following genotypes: WSA-1 (MATα lys2), WSA-70 (MATα ade2 lys2), WSA-591 (MATα ade2), WSA-1226 (MATα ura5), WSA-2126 (MATα ura5 cpk1Δ: : ura5), WSA-3002 (MATα/MATα fusant from WSA-1226×WSA-70×WSA-68), WSA-3019 (MATα haploid blastospore), WSA-3070 (MATα/MATα diploid blastospore recovered from JEC-21 hypha without basidiospore chains), WSA-3098 (MATα haploid blastospore recovered from JEC-21 hypha with basidiospore chains), WSA-3112 (MATα haploid blastospore recovered from hypha without basidiospore chains), WSA-3145 (MATα ura5 cpk1Δ: : ura5 CPK1: : NEOr). MATa strains included WSA-65 (MATa ura5 lys1 ade2) and WSA-68 (MATa ura5 ade2 lys2). Nocodazole (Sigma-Aldrich, St. Louis, MO) stock was prepared in DMSO at 1. 5 mM and then used to prepare different dilutions in YPD broth (YPD-0. 075 µm nocodazole, YPD-0. 15 µm nocodazole, YPD-0. 30 µm nocodazole, YPD-0. 60 µm nocodazole, YPD-1. 2 µm nocodazole). JEC-21 cells were added to 1. 5 ml nocodazole-YPD broth in 15 ml snap cap tubes (BD Biosciences, Franklin Lakes, NJ), at a final concentration of 1×106 cells/ml. Tubes were shaken at 200× RPM at 30°C for 24 hrs. Five µl of overnight culture were then dropped onto filament agar and incubated at 25°C for 5 days. The quantitative assay was performed by growing cells as above, and then plating cells onto filament agar as described in the quantitative monokaryotic fruiting assay. Pictures were taken with an Olympus SZX12 stereo microscope (Olympus, Center Valley, PA) at 2× magnification. To perform α-α cell fusions, 1×106 cells of complementary, auxotrophic, MATα strains were mixed, cultured on YPD agar at 37°C for 24 hrs, and then transferred onto filament agar plates. The plates were incubated at 25°C for 24 hrs. The mixture was scraped from the plate, suspended in 1. 0 ml sterile H2O, and then 200 µl of cells from this suspension were spread onto MIN agar plates, which were then incubated at 30°C for 4 days to screen for fusants. Assisted mating reactions, in which two MATα strains were induced to fuse by including a MATa helper strain, were performed according to previously described methods [19]. Plates were photographed at 0. 5× magnification. Cells were harvested, fixed, and stained with propidium iodide (Sigma-Aldrich, St. Louis, MO), and then sorted as described by Sia et al. [35]. Cell cycle analysis was performed using a BD FACSCalibur flow cytometer (Becton Dickinson Biosciences, Sparks, MD). CellQuest Pro software was used for cell collection, and data analysis was performed using ModFit and FlowJo. G1, S, and G2 phases were identified using the Dean-Jett-Fox mathematical model. G2-arrested cells were identified as described [25], [26], [36], [37], [38]. This population of cells typically shows a large cell phenotype, an increased proportion of cells with 2C DNA content when compared to controls, and a DAPI staining pattern that shows larger nuclei vs. smaller condensed nuclei of G2 phase cells (see below). Cell sortings were performed on a BD FACSAria III (Beckton Dickinson Biosciences) cell sorter and analyzed using BD FACSDiva 6. 1 software. Aliquots of sorted live cells were also used to perform the quantitative monokaryotic fruiting assay. All FACS analysis was performed at the Flow Cytometry Core Laboratory at The University of Texas Health Science Center at San Antonio. Cells were stained with DAPI according to the method described by Fuchs et al. [39]. Briefly, 1×107 cells were harvested from 24 h YPD agar plates, which were grown at either 30°C or 37°C, washed twice with phosphate buffer (0. 1 M KH2PO4,1. 25 mM EGTA, 1. 25 mM MgCl2 at pH 6. 9), then resuspended in 400 µl fixative (5% paraformaldehyde in wash buffer) followed by incubation at room temperature for 90 minutes. The cells were then washed three times in wash buffer and incubated with Lysing Enzymes (Trichoderma harzianum, Sigma-Aldrich, 1 mg/ml in sterile distilled water) for 20 min at 37°C. The cells were then washed once with sterile water, gently resuspended in 400 µl 0. 3% Triton X-100 (Sigma-Aldrich), and permeabilized by incubation at room temperature for 15 minutes. The suspension was then pelleted and washed three times with PBS. Cells were stained in DAPI (Sigma-Aldrich) (0. 1,0. 2, or 0. 5 µg/ml) for 15 minutes, washed twice with PBS, and then resuspended in 200 µl prior to visualization. Images were captured on a Zeiss AxioImager Z1 microscope (Carl Zeiss Microscopy, LLC, Thornwood, NY) equipped with an AxioCam MR3_2 CCD camera, using the filters for DAPI (365 nm excitation, 395 nm beam splitter, 420–470 nm emission filter) and differential interference contrast (DIC, Nomarski contrast). Image analysis and adjustments were performed using Axiovision software (Zeiss, Version 4. 8). Images were adjusted only for frame alignment (to overlay DAPI over DIC), brightness, and contrast (adjustment of +0. 01 contrast units over default), and all images received the same treatment.

Title: The Production of Monokaryotic Hyphae by Cryptococcus neoformans Can Be Induced by High Temperature Arrest of the Cell Cycle and Is Independent of Same-Sex Mating Summary: Fungi typically grow vegetatively as either yeast or hyphae. Many of the major human fungal pathogens can generate both morphologies and are referred to as the dimorphic fungi. Cryptococcus neoformans is a yeast-like fungus that has not been traditionally thought to be dimorphic since hyphae production typically occurs during the mating process between cells of opposite mating types. However, C. neoformans also can generate the hyphal state from haploid cells (called monokaryotic or haploid fruiting) in the absence of the opposite mating type. Recent studies have shown that the mechanism behind this process also involves mating, however, the mating reaction occurs between cells of the same mating type. Here we describe a unique mechanism responsible for monokaryotic fruiting that is independent of mating and does not proceed through a diploid intermediate. Instead, the key requirement for hyphal induction appears to be cell cycle arrest. Importantly, arrested cells display an enlarged cell phenotype, which has been observed in vivo in recent reports and has been hypothesized to be a novel protection strategy against host defenses. C. neoformans appears to have an extensive morphological repertoire, which likely contributes to its success as both a pathogen and a saprophyte.

lay_plos

en

Summarize: A teen has been indicted on manslaughter and assault charges following a May 2014 crash that left five teens dead and two adults injured. Cory Gloe, 18, of Farmingdale, was indicted on five counts of manslaughter, two counts of assault, reckless endangerment, five counts of criminally negligent homicide, leaving the scene of an incident and reckless driving. Police say a car crossed the center lane on a New York road and crashed head-on into a larger vehicle, killing four teenagers and leaving three other people with serious injuries. Scroll down for video... Facing justice: Cory Gloe of Farmingdale, Long Island, was allegedly involved in a street race that led to a crash that killed five and injured two has been indicted on manslaughter and assault charges. Charges: Gloe was charged with with assault, reckless driving and reckless endangerment. Thirteen of the charges in the 17-count indictment are felony offenses and could face 15 years behind bars. Sole survivor: It is alleged that just after midnight on May 10, 2014, Gloe, then 17 and possessing a full driver’s license was participating in organized street races on the streets behind as shopping mall. Investigators had been looking into whether drag racing was involved in the accident. The arraignment was attended by many friends and relatives of those killed, all of whom silently left the courtroom on Friday without responding to questions. Nassau County Court Judge Terence Murphy suspended Gloe's license and set bail at $50,000 cash or bond. He is due back in court on February 11, and if convicted of the top charge against him, he faces a maximum of five to 15 years in prison. Tragic scene: The incident happened just after midnight on Saturday, May 10 last year, on Conklin Street in Farmingdale. Tragedy: Cody Talanian (left) and his friend Tristan Reichle (right) were both killed after a car crash last May. Too soon: Carly Lonnberg (left) and Jesse Romero (right) also died shortly after the crash. Heartbreaking: Noah Francis (pictured) was also killed in the accident. According to detectives, a 2001 Nissan driven by a 17-year-old Tristan Reichle was traveling westbound when the vehicle crossed into the eastbound lanes and struck a GMC Suburban operated by a 53-year-old man. There were five people inside, two of whom were ejected. Four victims died immediately after the accident, while the fifth died at the hospital the next day. Prosecutors allege that Gloe stopped at a red light in the lane next to Reichle, challenging him to a race several times. When the traffic signal turned green, the two cars crossed Route 110 and were racing each other westbound on Conklin Street when Reichle lost control. Reichle was killed in the crash along with 18-year-old Jesse Romero, 14-year-old Carly Lonborg and 15-year-old Noah Francis. Cody Talanian, 17, was pronounced dead at 10 p.m. the following evening at Nassau University Medical Center. High schoolers: All five teens who were riding in this 2001 Nissan were from Farmingdale High School. Disbelief: The families of the teens awoke to the awful news on Mother's Day 2014. All five victims attended Farmingdale High School, though one was a former student. 'Today's indictment is the result of an event that brought heartbreak to these families, the innocent victims who were injured, and the Farmingdale community since last May,' Acting District Attorney Madeline Singas said. 'Unfortunately nothing will alleviate the pain or bring back the lives lost in this senseless crash. We owe it to our kids and everyone on our roads to remember that speed and racing kill. As parents, grandparents and teachers we cannot emphasize enough to our kids that when you are old enough to drive a car, you are also old enough to be held responsible for your decisions behind the wheel.' Gloe's attorney Stephen LaMagna insists his client will be acquitted, calling the incident 'unbelievably tragic,' and noted Gloe's vehicle was not involved in the actual crash. 'My client did not cause this accident in anyway and in any form,' LaMagna said. 'My client, his family, have been heartbroken as well as the rest of the community over what happened 9 months ago. Unfortunately many times in tragedies such as these, invariably the district attorney's office has a need to blame somebody, anybody whether or not that person was truly responsible for this tragedy.'

Summary: Five teenagers were killed in a car crash on Long Island when their Nissan crossed the center lane and smashed into another vehicle. Jesse Romero, Carly Lonborg, Tristan Reichle, Cody Talanian, and Noah Francis were all killed. Two people in the other vehicle were seriously injured. Cory Gloe faces up to 15 years in prison if convicted of the top charge against him. The judge suspended his license following his arraignment and he was freed on $50,000 bail.

cnn_dailymail

en

Summarize: Heavy snow has turned New England into a frozen winter wonderland, but the icicles and ice dams taking over local buildings are also a dangerous threat. Portland, Maine resident Adam Sousa had his car wrecked this weekend by falling ice that shattered the windows, dented the body and caved in the bumper. Luckily the 28-year-old wasn't in the car at the time, since a 3-foot-long ice chunk smashed through the windshield and landed right between the driver and passenger seats. He says the ice chunk was so heavy, both he and a tow truck driver struggled to move it together. Scroll down for video. Shattered glass: A resident in Portland, Maine had his car wrecked on Sunday when it was struck by falling ice in the Old Port part of town. Look up: The huge chunks of 3-foot-long ice, some weighing more than 100 pounds, hit 28-year-old Adam Sousa's car, seen parked above. Out of luck: Sousa's liability insurance doesn't cover the cost to fix his 2003 Chevrolet Cavalier. He is now working with the building's owner Joseph Soley to see if that man's insurance will help fix the car. 'I thought maybe an icicle would fall off, not a whole car-size piece of ice,' Mr Sousa told WMTW of his expectations parking his 2003 Chevy Cavalier outside the building on Sunday. Mr Sousa dropped his car off in the morning, took the bus to Freeport and then returned in the afternoon to find the startling damage. Unfortunately, his liability insurance doesn't cover the cost to fix his car and the city can't help either. 'What they told me when I spoke to the police is that they can't really do much, because it's an act of nature. But from what I saw, the size of those ice chunks, they're a 100- 150-pound ice chunks,' Mr Sousa said. However, he has contacted the building's owner, Joseph Soley, who is looking into whether his insurance might cover the cost of repairs. Mr Sousa told the Press-Herald that he talked to Mr Soley on Tuesday afternoon and that their conversation was friendly. Wrecked: Falling ice chunks shattered several windows on Mr Sousa's car and banged up much of the body. Rule breaking: It's a violation of Portland city ordinance to leave dangerous icicles and ice dams on buildings. 'He said he would contact his insurance and he seemed very nice about it,' Mr Sousa said. 'He did ask why I didn’t park somewhere else.' On Monday, the city investigated and determined that Mr Soley had violated a city ordinance against dangerous ice and snow buildup. Building owners are required to clean up such threatening ice, and if not, they can be fined up to $250. City officials say Mr Soley will not face any sort of criminal charges. 'When that ice dam is noticed, or obviously presents a hazard, they're required to mitigate that hazard.' Tim Nangle of Portland Fire and EMS said. Time of need: Friends have been helping drive Mr Sousa to and from his job as a bartender at a local restaurant and a GoFundMe campaign has already raised over $3,000 for Mr Sousa's expenses. However, Mr Nangle added that sometimes there are logistical challenges to keeping building owners from cleaning the dangerous ice in winter - such as the demand for workers to scrape the ice across the city. Still, Mr Nangle says he's never seen damage to a car as bad as what happened to Mr Sousa. 'It really was a significant chunk of ice that went through the windshield,' Mr Nangle told WCSH. 'I was surprised and actually glad that there was no one in the car.' In the meantime, friends have been driving Mr Sousa to and from his job as a bartender at DiMillo's Restaurant & Lounge, and he has encountered even more support through a crowd-funding campaign that has already raised over $3,000 dollars. The U.S. Bartenders Guild has also offered to host a fundraiser for Mr Sousa. 'The amount of help that everybody has been putting forth is just unbelievable,' Mr Sousa said

Summary: Adam Sousa, 28, parked his car in downtown Portland, Maine on Sunday and returned to find it wrecked by falling ice chunks. Some of the chunks that shattered the car's windows and dented the body were as long as three feet and weighted over 100 pounds. The city says building owner Joseph Soley violated an ordinance to keep his building clear of ice, which carries a $250 fine.

cnn_dailymail

en

Summarize: A Django Unchained actress who allegedly had sex with her boyfriend in a car has blamed society and the shooting of teenager Michael Brown for why she accused police of 'racially profiling' her. Daniele Watts claimed she was harassed by Los Angeles Police Department officers after they arrested her as she straddled her boyfriend Brian James Lucas in a silver Mercedes in Studio City. Watts, who is black, and her white partner were having sex in the vehicle near the CBS TV studios at the time, according to witnesses. However, the actress has maintained they were only kissing. Now, she has blamed the killing of 18-year-old Brown in the St Louis suburb of Ferguson - as well as her own history of allegedly racial profiling - for why she 'exploded' at police. Scroll down for video. Couple: Daniele Watts claimed she was harassed by officers of the Los Angeles Police Department, who arrested her as she embraced her boyfriend Brian James Lucas (both pictured) in a car in Studio City. 'Lewd conduct': Watts, who is black, and her white partner were having sex in the vehicle near the CBS TV studios at the time, according to witnesses. However, the actress has maintained they were only kissing. Speaking at a panel discussion at USC's School of Law on Monday, Watts said: 'If we're going to condemn me, then we also have to look at the entire society that I am a product of.' Referring to Brown's death at the hands of officer Darren Wilson on August 9, she added: 'At a certain point, enough is enough and somebody has to say something.' Watts also told listeners that witnessing her black father being profiled by a white policeman when she was 16 had influenced her to pull the race card on officers on September 11, according to the Los Angeles Times. The actress was detained by LAPD Sargaent Jim Parker after he responded to complaints that a couple was engaging in a 'lewd sex act' and she refused to provide identification when asked. Arrest: Now, Watts (pictured crying during her arrest) has blamed the killing of Michael Brown in the St Louis suburb of Ferguson - as well as her own history of allegedly racial profiling - for why she 'exploded' at police. Shooting: Referring to Brown's (left) death at the hands of officer Darren Wilson (right) on August 9, she told listeners at USC's School of Law: 'At a certain point, enough is enough and somebody has to say something' Although Watts was released with no charges at the time, a subsequent investigation'revealed witnesses who were willing to provide evidence of a criminal act,' said a department spokesman. Two weeks ago, the star and Lucas were charged with misdemeanor lewd conduct by the Los Angeles city attorney's office. The couple have said they are 'appalled' by the charges. As the case hit the headlines in September, Watts was lambasted for 'crying wolf' by numerous civil rights activists, including Los Angeles Urban Policy Roundtable President, Earl Ofari Hutchinson. Hutchinson, who initially supported Watts's racial profiling allegations but later changed his tune, told reporters: 'We take racial profiling very seriously. It's not a play thing. It's not trivial.' In an audio recording, Watts could be confronting police in a heated exchange during which she used the N-word, and claimed legal expertise from her experience playing a cop on TV. 'You're not the one in handcuffs, you're not the one who's spent your life being called a n*****, and growing up in the South and now I get the cops called on me,' she told officers through tears. 'I'm an actress at this studio! I'm in a major sitcom and I'm still being put in handcuffs because I'm making out with my boyfriend.' The release of the audio file came shortly after still images showed Watts - who played Coco in the 2012 film Django Unchained - straddling Lucas in a parked car in public. Allegations: Watts is perhaps best known for her small role in Django Unchained, playing a slave called Coco (left). She posted the picture on the right to Facebook, claiming she was injured in the September 11 arrest. Sgt Parker, a 25-year veteran of the LAPD, later said he was offended by claims that Watts was harassed because she is a black woman with a white boyfriend. He said he feared that if he and other officers had not recorded their run-in with Watts, her allegations would have ruined their careers. Watts was joined at the panel discussion by USC professors Camille Gear Rich, Michelle Gordon, Shana Redmond and Diana Williams. She reportedly hoped the event would lead to broader societal conversation about race. Brown's shooting in Missouri in August sparked violent demonstrations in the city of Ferguson, with many residents claiming the teenager was gunned down by Mr Wilson because of his race. A grand jury is considering the evidence from the incident. They are expected to reach a conclusion this month. Watts and Lucas are due to appear in court on November 13. If convicted, they could each face a maximum sentence of six months in jail and a $1,000 fine

Summary: Police stopped Daniele Watts and boyfriend after reports of lewd conduct. She accused officers of racially profiling her, saying they were just kissing. Now, she has blamed society and Michael Brown shooting for allegations. Said: 'If we're going to condemn me, we have to look at the entire society' Referring to Brown's death, added: 'At a certain point, enough is enough' Made comments at a panel discussion at USC's School of Law on Monday. Watts 'exploded' at LAPD officers after she was detained for not giving ID. Two weeks ago, she and partner charged with misdemeanor lewd conduct. If convicted, they could each face up to six months in jail and $1,000 fine.

cnn_dailymail

en

Write a title and summarize: The potential use of clinically approved beta-lactams for Buruli ulcer (BU) treatment was investigated with representative classes analyzed in vitro for activity against Mycobacterium ulcerans. Beta-lactams tested were effective alone and displayed a strong synergistic profile in combination with antibiotics currently used to treat BU, i. e. rifampicin and clarithromycin; this activity was further potentiated in the presence of the beta-lactamase inhibitor clavulanate. In addition, quadruple combinations of rifampicin, clarithromycin, clavulanate and beta-lactams resulted in multiplicative reductions in their minimal inhibitory concentration (MIC) values. The MIC of amoxicillin against a panel of clinical isolates decreased more than 200-fold within this quadruple combination. Amoxicillin/clavulanate formulations are readily available with clinical pedigree, low toxicity, and orally and pediatric available; thus, supporting its potential inclusion as a new anti-BU drug in current combination therapies. Buruli ulcer (BU) is a chronic debilitating mycobacterial disease of the skin and soft tissue. Although mortality is low, permanent disfigurement and disability are high, mainly affecting children and young adults. BU is found primarily in tropical regions of Africa, South America and the Western Pacific; however, it is also becoming a public health concern in some regions of Australia [1]. Before 2004, when the World Health Organization (WHO) published provisional guidance for the management of BU disease [2], antibiotics were viewed as relatively ineffective and surgery remained the mainstay of treatment for BU [3–5]. In the late 1990s and early 2000s, however, in vitro studies demonstrated anti-BU activity of some antibiotics used for the treatment of tuberculosis (TB) and other non-tuberculous mycobacteria [6–8], and in vivo studies the potential for combining two drugs to provide improved treatment outcomes [9–13]. Soon after, clinical evidence showed the effectiveness of a combination of rifampicin plus streptomycin when it was administered for at least 4 weeks [14], and that routine implementation of such a therapy was possible in the field [15,16]; however, the use of the injectable streptomycin is often associated with adverse events and it is restricted in the treatment of pregnant women and young infants. In addition, the lack of an efficacious oral treatment remained one of the main obstacles to decentralizing care at local level in rural areas. These limitations motivated the scientific community to evaluate alternative oral treatments and clinical studies demonstrated that fluoroquinolones [17] or clarithromycin [18–20] could also be used in combination with rifampicin and were associated with fewer side effects compared to the injectable streptomycin. Thus, on March 24th, 2017, WHO recommended full oral treatment of 8 weeks daily combination therapy of rifampicin-clarithromycin [21]. While recommended regimens (rifampicin plus streptomycin or clarithromycin) allow cure of small lesions (<5 cm in diameter) without surgery [15,18], controversy remains regarding the best surgery approach for large lesions (>10 cm) [22,23]. Intermittent drug administration using rifapentine, a rifampicin analog with longer half-life, instead of daily rifampicin, has been also proposed as a strategy to facilitate treatment supervision in the field [24]. However, M. ulcerans strains resistant to rifampicin have been isolated after experimental chemotherapy in mice [25] and a recent report described the emergence of M. ulcerans strains resistant to rifampicin and streptomycin in the clinic [26]; further experiments would be, however, needed to identify the genetic basis of such resistance patterns and confirm the emergence of resistance in M. ulcerans clinical isolates. Nevertheless, these reports should be a warning sign since no alternatives for rifampicin are currently available. WHO currently recommends only four drugs for the treatment of BU: rifampicin, streptomycin, clarithromycin and moxifloxacin [2]. It would be thus desirable to increase the number of drugs available to treat BU and to develop a new therapy that would reduce both duration of treatment and time to healing after therapy completion for all type of lesions and suitable for children and pregnant women. Drug discovery and development for neglected diseases is especially delayed due to lack of interest from the main scientific and industrial communities. To speed up the process in the BU field, we applied knowledge gathered in TB R&D drug repurposing programs [27–30] where we (and others [31]) showed that beta-lactams strongly increased the bactericidal and sterilizing properties of rifampicin [28]. Rifampin is the cornerstone drug for TB (and BU) therapy with a direct relation between dose increase and therapy efficacy [32] due to its bactericidal and sterilizing activity in a dose-dependent manner [33]. However, the current WHO recommended 10 mg/kg (600 mg daily) is not its optimal clinical dosage [34] and some recent studies suggest that it could be safely increased to 35 mg/kg daily for TB therapy with a bacteriological effect on time to culture conversion [32,35,36]. More recently, dose-ranging high-dose rifampicin studies using a murine model of M. ulcerans disease showed that shorter BU treatments might be also feasible [37], suggesting that synergistic partners could serve to improve rifampicin efficacy without compromising tolerability and toxicity. Beta-lactams are one of the largest groups of antibiotics available today with an exceptional record of clinical safety in humans [38]. Used for decades, they had been traditionally considered ineffective for the treatment of mycobacterial infections (mainly TB) due to the presence of a beta-lactamase (BlaC) and the hydrophobic nature of the mycobacterial cell envelope [39]. However, after a seminal publication describing the in vitro activity of meropenem plus clavulanate against multi-drug (MDR) and extensively drug resistant (XDR) strains of M. tuberculosis [40] and its anecdotal use in salvage therapies for XDR patients [41], the first study convincingly demonstrating the clinical efficacy of beta-lactams was recently published [29]. These studies provided evidence of their anti-mycobacterial clinical potential, opening a new avenue to identify new drugs and optimize current BU therapy. In this study, we are translating knowledge and concepts of drug repurposing and synergy generated in TB R&D programs to assess the potential inclusion of beta-lactams for BU therapy. We propose the combination of amoxicillin/clavulanate as a new anti-BU treatment in combination with current oral BU therapy, rifampicin and clarithromycin, with the potential of treatment shortening and readily implementation in the field. M. ulcerans strain NCTC 10417 (ATCC Number: 19423; Lot Number: 63210551) was used for initial screening assays. Further validation studies were performed with clinical isolates from different geographical origins: ITM 063846, Benin; ITM 070290, China; ITM 083720 and ITM C05143, Mexico; ITM 941327, ITM C05142 and ITM M000932, Australia; ITM C05150, DR Congo; ITM C08756, Japan, purchased from the Belgian Co-ordinated Collection of Micro-organisms (BCCM). M. ulcerans cells were initially grown at 30°C to an optical density at 600 nm (OD600) of 0. 5–1. 0 in tissue culture flasks containing 7H9 broth supplemented with 0. 2% glycerol, 10% OADC and 0. 05% (vol/vol) Tyloxapol. Aliquots of 500 μL were then stored at -80°C and the number of colony forming units (CFU) in the freeze stock enumerated. Every experiment was performed starting from a new frozen stock to avoid excessive passage of the original strain. Cells were also routinely passaged on Middlebrook 7H10 agar plates (Difco) supplemented with 10% (vol/vol) OADC to ensure purity of the isolate. Rifampicin (R3501-1G; Lot Number: SLBH7862V) and meropenem (M2574; Lot Number: 055M4705V) were purchased from Sigma. GlaxoSmithKline provided clarithromycin, streptomycin, clavulanate and all other beta-lactams used in this study. Minimal Inhibitory Concentrations (MIC) were determined in 7H9 broth supplemented with 0. 2% glycerol, 10% OADC and without Tyloxapol using triplicate two-fold serial dilutions of compounds in polystyrene 384- or 96-well plates. MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide] was used as the bacterial growth indicator [30,42]. For cell density calculations, a culture having an OD600 of 0. 125 was found to contain approximately 107 cfu/mL. Cultures were sampled (50 μL in 384-well plates or 200 μL in 96-well plates) at a final cell density of 106 cfu/mL and incubated at 30°C in the presence of the drug (or drug combinations) for 6 days before addition of 12. 5 μL (384-well plates) or 30 μL (96-well plates) of a MTT / Tween 80 (5 mg/mL / 20%) solution mix. After further overnight incubation at 37°C, OD580 was measured. The lowest concentration of drug that inhibited 90% of the MTT color conversion (IC90) was used to define MIC values. Checkerboard assays and calculations of the Fractional Inhibitory Concentration Index (FICI) were used to define the degree of pairwise drug interactions, as previously described [28] (for a visual representation and deeper understanding of the checkerboard assay refer to the supplementary information of Ramón-García et al. [30]). Up to quadruple combinations of rifampicin, clarithromycin, beta-lactams and clavulanate were also tested. For this, checkerboard plates were prepared with rifampicin in the y-axes, the beta-lactam in the x-axes, and clarithromycin added in the z-axes as fixed sub-MIC (1/2,1/4, and 1/8xMIC values) concentrations for every checkerboard plate; typically the 1/8xMIC plate was used for synergy calculations of the quad combos. When assayed, clavulanate was added at a fixed sub-MIC concentration of 5 μg/mL. Increase efficacy of compounds (synergistic MIC, MICsyn) when in combination (fold-MIC reduction) was always reported versus the activity of drugs alone. The Most Optimal Combinatorial Concentration (MOCC) was defined as the lowest possible concentration of every compound that, when assayed together, prevented bacterial growth, i. e. in an isobologram representation this would be the closest point to the axes intersection. Clinically approved beta-lactams representing different sub-families, i. e. meropenem (carbapenems), cephradine and cefdinir (cephems), faropenem (penems) and amoxicillin (penicillins) were assayed in vitro in a checkerboard format to assess their synergistic interactions with rifampicin in the absence and presence of clavulanate, a beta-lactamase inhibitor, against the M. ulcerans ATCC strain. A pattern of strong synergistic interaction was observed between rifampicin and all beta-lactams tested (Fig 1); however, no interaction was observed when the same assay was conducted using combinations of rifampicin and the currently WHO recommended anti-BU drugs, i. e. streptomycin, clarithromycin and moxifloxacin (Fig 1 and S1 Fig). Dose-response curves indicated that the activity of rifampicin (reflected in MIC reduction) was increased on average 16-32-fold (up to 128-fold in some cases) and, vice versa, the activity of the beta-lactams was strongly enhanced by rifampicin. In the case of amoxicillin, its activity was further increased 512-fold in combination with clavulanate (S2 Fig and S1 Table). Beta-lactams not only had synergistic interactions with rifampicin but also with clarithromycin, the second drug recommended as first-line anti-BU therapy. These results prompted us to test the inhibitory effect of double clarithromycin-beta lactam, and triple rifampicin-clarithromycin-beta-lactam combinations (Fig 2). Our results indicated that, when in double or triple combinations, much lower sub-inhibitory concentrations were equally potent at inhibiting M. ulcerans growth than the additive effects of the compounds alone. MIC values were also lower than in other pairwise combinations. For example, the MIC of amoxicillin was greater than 32 μg/mL; however, its synergistic MIC (MICsyn) was reduced to 1 μg/mL in the presence of rifampicin, to 0. 25 μg/mL when clavulanate was also added, or to 0. 062 μg/mL when both clavulanate and clarithromycin were included together with rifampicin, i. e., an MIC reduction of ca. 500-fold for amoxicillin when in the quadruple combination. Similar results were obtained for combinations of meropenem or faropenem and rifampicin, with MIC reductions as high as 80-fold when tested within triple combinations. In these assays, clarithromycin was added at a fixed concentration of 1/8 its MIC value alone, being its presence critical to achieve the multiplicative effect observe in the quadruple combinations. Our initial discoveries described above were performed using M. ulcerans ATCC 19423, a reference strain that does not produce mycolactone, a polyketide-derived macrolide secreted by M. ulcerans and responsible for the tissue damage pathology in Buruli ulcer [43]. In order to validate the pattern of synergistic interactions between rifampicin, clarithromycin and beta-lactams and to gauge the potential of introducing a beta-lactam in the treatment of BU, we expanded our analysis to a collection of M. ulcerans clinical isolates from different geographical locations and focused our synergy interaction studies on the amoxicillin /clavulanate combination. When in the quadruple combinations, the activities of all three drugs (clavulanate was added at a fixed 5 μg/mL concentration, more than 20-fold less its MIC) were strongly enhanced; depending on the strain tested, these interactions could range from ca. 5 to 600-fold (rifampicin), ca. 4 to 2,000-fold (amoxicillin) and ca. 20 to 80-fold (clarithromycin) (Table 1). Every possible pair-wise and triple combination was also evaluated showing strong synergism between amoxicillin and both rifampicin and clarithromycin, but not between rifampicin and clarithromycin, similar to previously described for the ATCC strain. Clavulanate enhanced the activity of amoxicillin (as expected) but had no effect over clarithromycin and only minor enhancements (in some cases) over rifampicin (S3 Table). Amoxicillin is inactivated by beta-lactamase enzymes, limiting its clinical use. In fact, dose-response studies demonstrated that amoxicillin alone was not active (MIC > 16 μg/mL); however, a strong shift in the dose-response curve was observed by adding clavulanate and this shift was further enhanced when clarithromycin and rifampicin were also present in the combination at sub-MIC concentrations (Fig 3), thus confirming our MIC/synergy data and the potential of amoxicillin/clavulanate as a new anti-BU therapy, both alone and in combo with rifampicin, clarithromycin or both. We have in vitro characterized the antimicrobial interactions of rifampicin with anti-BU drugs and beta-lactams and found that, while there was no synergy between rifampicin and current anti-BU drugs, it had strong synergistic interactions with beta-lactams. What is more, beta-lactams also displayed synergism with clarithromycin, the other first line drug in BU therapy. Our studies confirmed previous data showing lack of in vitro interaction between rifampicin and clarithromycin [44], thus reinforcing synergy data observed with beta-lactams. Similar observations of no interaction between rifampicin and clarithromycin were also recently described in murine models of M. ulcerans infection [37] Working with M. ulcerans is challenging due to its slow generation time (ca. 48 hours), even slower than M. tuberculosis. Current methodologies to perform susceptibility testing against clinical isolates are often time consuming and cumbersome (use of agar proportion methods), thus, requiring several months to generate results [6,8, 26]. Improvements have been introduced using luminescent reporter strains [45]; however, this technology is limited to specific engineered strains and cannot be widespread applied to clinical isolates. Here, we were able to perform synergy studies, obtaining results in just seven days, in a medium-throughput manner against a panel of clinical isolates by adapting a methodology previously described for TB research and already validated to determine MIC values in slow growing mycobacteria [28]. Similar redox-based assays (using alamar blue) have been previously employed in antimicrobial discovery programs targeting M. ulcerans [46,47]. This methodology could provide a cost and time effective assay to implement in the clinical practice in Buruli ulcer drug resistance surveillance campaigns. Over the last decade, BU therapy has undergone a tremendous improvement with the introduction of chemotherapy; however, there is only a limited number of drugs recommended by WHO for BU therapy, namely rifampicin, streptomycin, clarithromycin and moxifloxacin [2]. Current WHO recommended therapy is a fully oral eight weeks daily regimen with a combination of rifampicin and clarithromycin [20]. Although effective and mostly well tolerated, combination treatment of rifampicin plus streptomycin or clarithromycin is associated with undesirable side effects that might include mild (anorexia, nausea, abdominal pains and altered taste) or severe (deafness, skin rashes, jaundice, shock, purpura or acute renal failure) symptoms [2]. In a scenario where administration of any of these drugs needs to be interrupted, therapeutic options would remain limited to the use of moxifloxacin, an antibiotic contraindicated during pregnancy and within the pediatric population. A similar situation would occur in the eventual development of resistance to any of these drugs, especially rifampicin. Since they are administered in pairwise combinations, this would effectively imply monotherapy, further promoting the emergence of resistance. This is similar to another mycobacterial disease, leprosy, for which this threat was largely ignored and just recently WHO issued guidelines including procedures for the detection of drug resistance [48]. Thus, an alternative drug regimen would be required to treat resistant M. ulcerans strains [26], even though the emergence of resistance in M. ulcerans might follow different dynamics compared to M. tuberculosis and M. leprae (environment vs. host reservoirs, respectively) and clinical resistance has not been conclusively demonstrated to date. BU is a neglected disease mainly affecting rural areas in under-resourced countries where medicine access and logistics might prove difficult, and hospitalization and loss of income for patients and families might compromise patient’s adherence to the 8-weeks antibiotic course. A shortened, highly effective, all-oral regimen is urgently needed to improve care for this neglected tropical disease; this would reduce indirect costs and barriers to therapy. The history of TB chemotherapy teaches us that combination therapy is critical for optimal cure outcome and treatment shortening [49]. Translation of this knowledge into BU therapy suggests that more drugs need to be added to the current rifampicin-clarithromycin combination in order to improve and shorten the duration of treatment. Most beta-lactams tested in this study were active against M. ulcerans and enhanced the anti-BU activity of rifampicin to different degrees. Cephradine is a first-generation cephalosporins developed in the 1960s, also recently described to be active in vitro against M. tuberculosis [28]; however, cephradine was long ago discontinued and access to other first-generation cephalosporins, such as cefadroxil or cephalexin, is limited in many countries. Cefdinir is a third-generation cephalosporin active against pneumonia, skin and soft tissue infections, although with low oral absorption [50]. It is currently used in the clinic, widely distributed and access to it could be readily available; however, its synergistic profile with rifampicin was weaker compared to other beta-lactams (S1 Table). Although meropenem was active against pulmonary TB in a recent clinical study [29], it needs to be administered intravenously, not a practical approach in under-resourced countries where oral drugs are required. Faropenem, an orally administered beta-lactam, did not show activity in the same clinical trial due to the low drug exposure in plasma after oral administration [29]. Finally, amoxicillin/clavulanate showed good activity and very strong synergistic interaction with rifampicin. The combination of amoxicillin plus clavulanate is a broad-spectrum antibacterial available for clinical use in a wide range of indications and is now used primarily in the treatment of community-acquired respiratory tract infections [51]. It was first launched in the UK in 1981; by the end of 2002, it was clinically available in various formulations in over 150 countries around the world. In addition to high efficacy, it has a well-known safety and tolerance profile, including for pregnancy and pediatric used, based on over 819 million patient courses worldwide, with the main contraindication being allergy to penicillin derivatives. Disruption of the gut microbiota is the main side effect of long-term use of amoxicillin/clavulanate, mainly caused by the presence of clavulanic acid in the formulation [52]. For TB treatment, amoxicillin/clavulanate is included in Group 5 (anti-TB drugs with limited data on efficacy and long-term safety in the treatment of drug-resistant TB) of the WHO 2011 TB drugs classification and in Group D3 (add-on agents, not core MDR-TB regimen components) of the WHO 2016 MDR-TB drugs classification [53]. In 1983, Cynamon et al. reported the in vitro bactericidal activity of amoxicillin/clavulanate against 15 isolates of M. tuberculosis, at concentrations of amoxicillin lower than 4 μg/mL [54], and some years later, Nadler et al. case reported the effective treatment of MDR-TB patients with the addition of amoxicillin/clavulanate to the second-line therapy [55]. Two contradictory follow up clinical studies, 2-days Early Bactericidal Activity (EBA), suggested that its activity was comparable to that reported for anti-TB agents, other than isoniazid [56], but also questioned its role in the treatment of tuberculosis [57]. The dosing interval of the amoxicillin/clavulanate therapy might explain these differences; while in the first EBA it was divided into three daily doses, it was given as a single high dose in the second one. More recently, a 14-days EBA study demonstrated activity of a combination of meropenem plus amoxicillin/clavulanate [29]; it remains to be determined whether this activity was due to any of the components alone or the combination therapy as a whole [58]. In fact, in vitro studies have demonstrated synergistic interactions among amoxicillin and meropenem (and other beta-lactams), rifampicin and ethambutol against M. tuberculosis [28,58–60]. In our in vitro assays with M. ulcerans clinical isolates, we found that amoxicillin had no activity (typically MIC values > 16 μg/mL) but that its MIC could be reduced to 1 μg/mL in the presence of clavulanate (S3 Table), similar to previously reported to the closely related M. marinum species and other non-tuberculosis mycobacteria [61]. It also displayed strong synergistic interactions with rifampicin and clarithromycin and, in quadruple combinations, its activity was enhanced up to 2,000-fold in some cases, with average MIC ranges between 0. 031 to 0. 25 μg/mL (Table 1). For infections caused by other bacterial pathogens, susceptibility breakpoints of amoxicillin/clavulanate are established at ≤ 2 μg/mL (or ≤ 4 μg/mL for high-dose formulations) and mean peak plasma concentrations of amoxicillin range from 7. 2 to 17 μg/mL, depending on the formulation [52], well above the synergistic MIC values reported in this work. The bacteriological efficacy of penicillins is dependent on the time its free plasma concentration remains above the MIC (time over the MIC value, fT>MIC). For other bacterial infections, it has been estimated that a fT>MIC of ca. 30–40% of the dosing interval is required for bactericidal activity [62]. In the case of M. ulcerans, this target therapy could be achieved using standard amoxicillin/clavulanate formulations of 500/125 mg (4: 1) or 875/125 mg (7: 1) administered three times a day, or the high-dose extended release formulation of 2000/125 (16: 1) that would allow administration twice a day [52], an important consideration for treatment compliance in under-resourced settings. Thus, according to our in vitro data, amoxicillin/clavulanate could have an important role in the treatment of BU alone and, more importantly, in combination with current first-line anti-BU therapy since no pharmacological drug-drug interactions are described among amoxicillin/clavulanate and rifampicin or clarithromycin [51]. But, what could be the benefit of adding amoxicillin/clavulanate to the current anti-BU therapy? Besides being able to treat secondary infections associated with BU lesions, it has been proposed that the median time to healing is related to the bacterial load in the lesions at the beginning of therapy and the presence of persister bacteria [63]; in fact, healing of up to two thirds of patients occurs within 25 weeks from the start of treatment but for some patients this can take up to a year. One of the reasons for this slow healing could be due to a high initial bacterial load. In fact, active infection late into the recommended 8-week course of antibiotic therapy could be found in slowly healing lesions [22,64]. Extensive histopathologic studies also demonstrated that M. ulcerans is essentially confined in extracellular areas of necrosis in skin [6]. Under this circumstances, amoxicillin/clavulanate would be extremely effective at targeting extracellular bacteria with rapid bactericidal activity, thus reducing initial bacterial burden, local levels of the immune-suppressive mycolactone toxin, and allowing local recovery of the host immune response to clear remaining bacteria. What is more, in vitro studies have demonstrated the sterilizing activity of synergistic combinations of beta-lactams and rifampicin [28], which could target those remaining persistent populations, thus shortening treatment and healing times. Rapid bacterial killing would also imply a reduction in the risk of development of resistance; even in the scenario of infections caused by bacteria resistant to rifampicin, this could still be re-introduced for BU therapy if it was administered with amoxicillin/clavulanate, as previously demonstrated in M. tuberculosis [28]. Finally, because of their synergistic interactions with clarithromycin, it could replace rifampicin in the treatment of HIV patients under anti-retroviral therapy. Our study comes as well with some limitations. First, although representative of different geographical origins, we only tested one ATCC strain and 9 clinical isolates. Further in vitro studies with a larger set of M. ulcerans clinical isolates would be needed to assess the full clinical potential and coverage of a triple combination including rifampicin, clarithromycin and amoxicillin/clavulanate. Second, our results were generated using synergy assays based on MIC determinations. This implies two limitations: (i) although an established approach in antimicrobial synergy assays, the activities of drugs alone and in combination were determined at a single time point after seven days of drug exposure and, (ii) synergy calculations inherently rely on sub-MIC concentrations, instead of actual serum levels achieved by drugs in clinical therapy. In order to thoroughly assess the effect of an eventual rifampicin, clarithromycin and amoxicillin/clavulanate combination at therapeutic concentrations, time kill assays would be required. However, even these assays would be challenging; since serum levels would be much higher than MIC values, the individual activity of drugs would mask any synergy signal using bactericidal activity as endpoint readout. Under these circumstances, and the much longer generation time of M. ulcerans compared to M. tuberculosis, assessing the sterilization capacity of such combinations would require extensive (months) incubation times of M. ulcerans cultures. Nevertheless, time kill assays are static pharmacokinetic (PK) / pharmacodynamics (PD) models where drugs are only added at the beginning of the assays and do not reproduce clinical therapy. The hollow fiber system, a dynamic PK/PD model by which posology and length of treatment can be mimicked in vitro, might provide data with higher prediction potential of treatment outcomes. This technology has proven a useful tool in the TB field, recently endorsed by the European Medicines Agency [65]; however, to date no laboratory has reported work on M. ulcerans using the hollow fiber system. Reasons for this might include the difficulties of working with a BSL3 pathogen that forms colonies in 2–4 months. Finally, in the field of BU (and TB), it is common practice to perform preclinical evaluation of drugs (or drug combinations) identified by in vitro studies using murine models of M. ulcerans infection. Although promising, experience from TB research has revealed inconsistencies between murine model data and clinical predictability [66,67]. In addition, mice are a sub-optimal in vivo model to evaluate the activity of beta-lactams since their pharmacokinetics and efficacy in mice do not predict those found in humans [68]; this is in part due to the fact that mice express an enzyme that degrades beta lactams (renal dehydropeptidase I, DPH-I) at levels that are several orders of magnitude higher than in humans [69,70], thus effectively reducing the time beta-lactams are over the MIC value. As such, murine models might not be the most appropriate development strategy for the use of beta-lactams in BU therapy. Because amoxicillin/clavulanate is a well-known antimicrobial with a clear track record of safety over decades of use, we believe that direct evaluation in clinical trials would be the fastest route to improve treatment of BU patients. In summary, using a repurposing approach and in vitro technology already developed in TB R&D programs, we have identified amoxicillin/clavulanate as a new potential anti-BU drug to be used alone or in combination therapy with rifampicin and clarithromycin, current first-line anti-BU drugs, with the potential to reduce length of therapy and time to healing. Based on the strong synergistic interactions among amoxicillin with rifampicin and clarithromycin, amoxicillin alone might be added to the full course of a shorter therapy. However, because the main role of amoxicillin/clavulanate in the anti-BU therapy would be to reduce the initial bacterial load found in the lesions, we propose the use of high-dose extended release formulations during the first two weeks of therapy.

Title: Triple oral beta-lactam containing therapy for Buruli ulcer treatment shortening Summary: Buruli ulcer (BU) is a chronic debilitating disease of the skin and soft tissue, mainly affecting children and young adults in tropical regions. Before 2004, the only treatment option was surgery; a major breakthrough was the discovery that BU could be cured in most cases with a standard treatment that involved 8 weeks of combination therapy with rifampicin and streptomycin. However, the use of streptomycin is often associated with severe side effects such as ototoxicity, or nephrotoxicity. More recently, a clinical trial demonstrated equipotency of replacing the injectable streptomycin by the clarithromycin, which is orally available and associated with fewer side effects. BU treatment is now moving toward a full orally available treatment of clarithromycin-rifampicin. Although effective and mostly well tolerated, this new treatment is still associated with side effects and only moxifloxacin is additionally recommended by WHO for BU therapy. New drugs are thus needed to increase the number of available treatments, reduce side effects, and improve efficacy with treatments shorter than 8 weeks. In this work, we describe for the first time the potential inclusion of beta-lactams in BU therapy. More specifically, we propose the use of amoxicillin/clavulanate since it is oral, suitable for the treatment of children, and readily available with a long track record of clinical pedigree. Its inclusion in a triple oral therapy complementing current combinatorial rifampicin-clarithromycin treatment has the potential to counteract resistance development and to reduce length of treatment and time to cure.

lay_plos

en

Summarize: La Roma è l'unica italiana arrivata nelle semifinali delle competizioni europee. I giallorossi si sono qualificati per le semifinale dell'Europa League 2020-2021 dopo aver eliminato l'Ajax in un doppio confronto complicato ma che ha visto i ragazzi di Paulo Fonseca avere la meglio grazie alla vittoria in trasferta alla Johan Cruijff Arena di otto giorni fa. Ai microfoni della BoboTV, Daniele Adani ha commentato così la partita di ritorno dei quarti di finale della squadra capitolina: "Tra le 8 di Champions e le 8 di Europa League non c'era un allenatore italiano, abbiamo parlato tante volte della Roma di Paulo Fonseca. La Roma di mister Fonseca contro l'Ajax ha fatto le sue due peggiori partite in assoluto, non è mai stata la vera Roma di Paulo Fonseca e i giallorossi sono quindi andati in semifinale non per meriti loro, ma a causa di alcune situazioni di gioco". In merito alla discussione sul valore delle due rose in campo ieri; l'ex difensore di Inter, Empoli, Brescia e Fiorentina ha affermato: "Chi ha detto che l'Ajax è superiore alla Roma? Quante cazzate, la Roma è più forte dell'Ajax, non scherziamo. Non si può preparare una partita rinunciando all'attacco, mai. Puoi controllare il possesso avversario senza la palla ma mai senza l'idea di attaccare, sia perché non è nell'idea di calcio di Fonseca e non è nell'idea del calcio".

Summary: La Roma di Paulo Fonseca si è qualificata per le semifinali dell'Europa League 2020-2021 e affronterà il Manchester United in un doppio confronto di fuoco. Per Daniele Adani, ex calciatore e attuale commentatore di Sky, i giallorossi sono passati non giocando bene: "La Roma di mister Fonseca contro l'Ajax ha fatto le sue due peggiori partite in assoluto, i giallorossi sono quindi andati in semifinale non per meriti loro, ma a causa di alcune situazioni di gioco".

fanpage

it

Write a title and summarize: Chronic Chagas disease cardiomyopathy (CCC) is an inflammatory dilated cardiomyopathy with a worse prognosis than other cardiomyopathies. CCC occurs in 30 % of individuals infected with Trypanosoma cruzi, endemic in Latin America. Heart failure is associated with impaired energy metabolism, which may be correlated to contractile dysfunction. We thus analyzed the myocardial gene and protein expression, as well as activity, of key mitochondrial enzymes related to ATP production, in myocardial samples of end-stage CCC, idiopathic dilated (IDC) and ischemic (IC) cardiomyopathies. Myocardium homogenates from CCC (N = 5), IC (N = 5) and IDC (N = 5) patients, as well as from heart donors (N = 5) were analyzed for protein and mRNA expression of mitochondrial creatine kinase (CKMit) and muscular creatine kinase (CKM) and ATP synthase subunits aplha and beta by immunoblotting and by real-time RT-PCR. Total myocardial CK activity was also assessed. Protein levels of CKM and CK activity were reduced in all three cardiomyopathy groups. However, total CK activity, as well as ATP synthase alpha chain protein levels, were significantly lower in CCC samples than IC and IDC samples. CCC myocardium displayed selective reduction of protein levels and activity of enzymes crucial for maintaining cytoplasmic ATP levels. The selective impairment of the CK system may be associated to the loss of inotropic reserve observed in CCC. Reduction of ATP synthase alpha levels is consistent with a decrease in myocardial ATP generation through oxidative phosphorylation. Together, these results suggest that the energetic deficit is more intense in the myocardium of CCC patients than in the other tested dilated cardiomyopathies. Chagas disease is a significant cause of morbidity and mortality in Central and South America, affecting about 13 million people [1]. The disease is caused by infection with the intracellular protozoan parasite Trypanosoma cruzi. About 30 % of infected patients develop chronic Chagas disease cardiomyopathy (CCC), an inflammatory cardiomyopathy that occurs decades after the initial infection. One-third of CCC patients further progress to a particularly aggressive, life-threatening dilated cardiomyopathy; CCC is a major indication of heart failure in Latin America [2], [3]. Clinical progression, length of survival and overall prognosis are significantly worse in CCC patients when compared to patients with dilated cardiomyopathy of non-inflammatory etiology, like idiopathic dilated or ischemic cardiomyopathies (IDC or IC, respectively) [4]–[7]. Due to migration from endemic countries, an estimated 300,000 people with Chagas disease are living in the USA, where a significant number of cases of CCC are expected per year [8]. The pathogenesis of CCC is unclear, and multiple mechanisms have been proposed (Reviewed in [9]). The most characteristic histopathological lesions in cardiac patients with CCC are consistent with inflammation and a myocardial remodeling process: T cell/macrophage-rich myocarditis, hypertrophy, and fibrosis with cardiomyocyte damage [10], [11]. The local cytokine production profile shows a T1-type response, with interferon-gamma-induced chemokines [12]–[15]. As the currently licensed anti-T. cruzi drugs may not be effective in preventing the progression of heart lesions of CCC [16], treatment is only supportive. In patients with refractory heart failure, the only available treatment is heart transplantation [17]. The absence of alternative treatment for CCC is a consequence of limited knowledge about the pathogenesis. Energy metabolism imbalances have been reported in dilated cardiomyopathies and heart failure [18]. Since the heart consumes more energy than any other organ, impairments in energy production could lead to a mechanical failure of the heart, and disturbances in electrical conduction [18]. Mitochondrial oxidative phosphorylation is essential for the production of energy for cardiac function. This system comprises the oxidative phosphorylation complex, which includes the electron transport chain (complexes I to IV) and the F1FO ATP synthase (complex V). In aerobic tissues, most ATP is synthesized via the mitochondrial F1FO ATP synthase complex. Studies have shown that certain components of oxidative phosphorylation may be impaired in heart failure [18]. Patients with IDC or IC show a reduced myocardial activity of complex III when compared to controls [19]. In IDC patients, a decreased activity of myocardial cytochrome c oxidase (complex IV) was observed [20]. With the progression of dilated cardiomyopathy, higher levels of spatial and functional heterogeneity within mitochondrial populations are observed, indicative of mitochondrial damage [21]. It has been reported that mitochondrial damage leads to loss of mitochondrial function, impairing energy production and cell physiology, and to the enhancement of pathologic function, producing oxidative-, calcium-, apoptosis-mediated myocyte injury [22]. Creatine kinases (CK) are also key enzymes of energy metabolism, which connect mitochondrial ATP-producing and cytosolic ATP-consuming process, and are thus of central importance to the cellular energy homeostasis [23]. This system acts as an energy buffer, in which mitochondrial creatine kinase (CKMit) catalyzes the transfer of high energy phosphate bond from ATP to creatine to form phosphocreatine and ADP. Phosphocreatine, a molecule smaller than ATP, diffuses rapidly from the mitochondria to the myofibrils, where myofibrillar creatine kinase (MM, MB, BB dimers, formed by CKM, the muscular isoform, and CKB, the brain isoform) catalyzes ATP production from phosphocreatine, generating free creatine, which diffuses back into mitochondria [18], [23]. Impaired ATP transfer and utilization may limit contractile function by means of a decrease in the average cytoplasmic ATP concentration [18]. Most of the components of the CK system are down-regulated in heart failure, with levels of creatine, phosphocreatine, CKMit and CKM all significantly reduced in animal models and in humans [24], [25]. CK deficiency in isolated hearts may cause a decline of over 70 % in ATP delivery to myofibrils, leading to a blunted contractile reserve [26]. Proteomic profiling of myocardium from CCC patients revealed that 27 % of identified proteins belong to energy metabolism pathways [27]. Using gene expression profiling, our group found differential expression of a significant number of genes involved in oxidative phosphorylation and lipid catabolism in myocardial samples from CCC patients, but not in samples from patients with dilated cardiomyopathy, when compared to samples from subjects without cardiomyopathy [15]. Genetic profiling studies showed that hearts of T. cruzi-infected mice have shown a decreased expression of oxidative phosphorylation enzymes [28]. Likewise, biochemical and histochemical analysis revealed a reduced activity of the respiratory chain complexes in hearts of T. cruzi-infected mice [29]. Proteomic analysis of myocardial samples from acutely T. cruzi-infected Syrian hamsters showed an up-regulation of the energy metabolism proteins glutamate oxaloacetate transaminase 1 and pyruvate dehydrogenase β, that may be associated with a high ATP demand after T. cruzi infection [30]. Inflammatory cytokines, which are present in the CCC myocardium and induce local signaling [12]–[15], have been reported to alter the energy metabolism. IFN-gamma was shown to inhibit the mitochondrial oxidative metabolism [31] and increase the rate of cardiac ATP depletion in cardiomyocytes [32]. Additionally, studies with cultured human skeletal muscle cells demonstrated that IFN-gamma treatment could inhibit CK activity [33]. Thus, the evidence is consistent with the hypothesis that the myocardium of patients with CCC could present an impaired energy metabolism. In order to test this hypothesis, we compared the protein and mRNA expression of CKM, CKMit, and the alpha and beta subunits of the catalytic F1 domain of ATP synthase complex (ATPα and ATPβ, respectively) in myocardial samples from CCC, with that of dilated cardiomyopathies of other etiologies, and healthy hearts from organ donors. We also measured total creatine kinase enzymatic activity in the same sample groups. Myocardial samples were obtained from left ventricular-free wall heart tissue from end-stage heart failure patients at the moment of heart transplantation. Samples from 5 CCC (at least 2 positive results in 3 independent anti-T. cruzi serology tests –ELISA immunoassay, indirect immunofluorescence assay and indirect hemagglutination test), 5 IDC (dilated cardiomyopathy in the absence of ischemic disease, negative serology for Chagas disease) and 5 coronary angiography-proven IC patients were collected (Table 1). Left ventricular free wall samples were also obtained from healthy hearts of organ donors, which were not used for transplantation for technical reasons. The protocol was approved by the Institutional Review Board of the University of São Paulo School of Medicine and written informed consent was obtained from the patients. Samples were cleared from pericardium and fat, quickly frozen in liquid nitrogen and stored at −70°C. Protein homogenates were obtained using lysing solution (1∶10 w/v) containing 7 mol/L urea, 10 mmol/L Tris, 5 mmol/L magnesium acetate and 4 % CHAPS, pH 8. 0, by mechanical homogenization (PowerGen, Fisher Scientific). For experiments measuring the creatine kinase enzyme activity, 20 mg of tissue was lysed in solution (1∶20 w/v) containing 0. 32 mol/L sucrose, 10 mmol/L HEPES and 1 mmol/L EDTA, pH 7. 4, by mechanical homogenization. The homogenate was then sonicated for three cycles of 10 s each to 10 Watts (60 Dismembrator Sonic, Fisher Scientific), centrifuged at 12,000 g for 30 min. Supernatants were collected and stored at −70°C. Protein quantification was performed with the Bradford method (BioRad). The samples from myocardial tissue (left ventricular free wall) were fixed in buffered formalin solution (pH 7. 2), embedded in paraffin, and cut into 5 µm sections. Sections were stained with hematoxylin-eosin (H&E) and picrosirius red. Extracts of myocardial samples containing 30 µg of protein were heated for 5 min at 95°C, and subjected to one-dimensional electrophoresis (SDS-PAGE) using 12. 5 % polyacrylamide gel and the vertical electrophoresis system Ruby SE600 (GE Healthcare). After electrophoresis, proteins were transferred from gel to a nitrocellulose membrane using the TE Semi-Dry Transfer Unit (GE Healthcare). The nitrocellulose membranes were incubated with monoclonal antibodies to proteins involved in energy metabolism: anti- F1FO ATP synthase alpha (ATPα) and anti- F1FO ATP synthase beta (ATPβ) (Molecular Probes), anti-mitochondrial creatine kinase (CKMit) and anti- muscle creatine kinase (CKM) (Santa Cruz) and polyclonal anti- glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (R & D Systems). Each membrane was subjected to incubation with compatible secondary antibodies conjugated with peroxidase, developed using ECL Plus Western Blotting Detection Reagents (GE Healthcare) and detection using X-ray equipment. Analysis of densitometry was performed using the program ImageQuant TL (GE Healthcare). Total RNA from left ventricle samples was isolated using the RNeasy Fibrous tissue kit (Quiagen). Contaminating DNA was removed by treatment with RNase-free DNase I. cDNA was obtained from 5 µg total RNA using Super-script II™ reverse transcriptase (Invitrogen). mRNA expression was analyzed by real-time quantitative reverse transcriptase (RT) -PCR with SYBR Green I PCR Master Mix (Applied Biosystems) and 250 nM of sense and anti-sense primers using the ABI Prism 7500 Real Time PCR System (Applied Biosystems). The following primers were designed using Primer Express software version 3. 0 (Applied Biosystems): GAPDH (M33197): (F) 5′-TGGTCTCCTCTGACTTCAACA-3′, (R) 5′-AGCCAAATTCGTTGTCATACC-3′; ATPα (NM_001001937): (F) 5′-TCTTCAAAAGACTGGGACTGCTGA-3′, (R) 5′-AAGACACGCCCAGTTTCTTCAAG-3′; ATPβ (NM_001686): (F) 5′-GCCCAGCATTTGGGTGAGA-3′, (R) 5′-GATTGGTGCACCAGAATCCAGT-3′; CKM (NM_001824): (F) 5′-GCTCTCTGTGGAAGCTCTCAACA-3′, (R) 5′-GATGAGCTGCTGCTGCTCCT-3′; CKMit (NM_001825): (F) 5′-TGACGAGGAGTCCTATGAGGTGTT-3′, (R) 5′-AGATCCGTTGTGTGCTTCATCAC-3′. After every PCR, an amplicon melting point curve was obtained. This yielded a single peak with the expected temperature provided by Primer Express software, confirming the specificity of the PCR. GAPDH mRNA expression was used for normalization. The amount of mRNA in the left ventricles samples was calculated using the 2-ΔCt method [34]. The enzymatic activity measurements of CK in the myocardial samples were performed using the CK-NAC kit (Doles). Basically, this method is a kinetic system where CK catalyzes the transphosphorylation reaction of ADP to ATP. A series of coupled enzymatic reactions produce NADH in a concentration directly proportional to the enzymatic activity of CK in the sample. The analyses were performed using a UV/Vis U-2001 spectrophotometer (Hitachi), monitoring the increase in absorbance of NADH per minute at the wavelength of 340 nm at 37°C, using a thermostatic bath (MultiTemp III, GE Healthcare). The measurement of enzymatic activity is given in international units (U); one unit of CK is the amount of enzyme that oxidizes 1 µmol/L of NADH per minute. Values were normalized by the amount of protein present in the sample. All statistical analyses were performed with GraphPad Prism 4. 0 software (GraphPad Software). Descriptive statistics are given as average and standard deviation. The non-parametric Newman-Keuls test was used for comparison between the groups. P-values <0. 05 were considered as statistically significant. While myocardial sections from all 3 cardiomyopathy groups displayed cardiomyocyte hypertrophy and fibrosis upon histopatholological analysis, myocarditis associated with a predominant lymphocytic infiltration was only observed among CCC heart lesions (Figure 1, Table 1). No significant differences were found in age, ejection fraction (EF) or left ventricular diastolic diameter (LVDD) among the three cardiomyopathy groups. Figures 2 A and B show the differential protein expression of the ATP-synthase, subunits alpha (ATPα) and beta (ATPβ), respectively. Representative immunoblots are depicted in Figure S1. The ATPα was 18 % less expressed in CCC myocardium when compared to myocardial samples from individuals without cardiomyopathies (p<0. 01). In contrast, IC myocardium showed an increase of 25 % (p<0. 001) in ATPα when compared to control samples, while in IDC there was no significant reduction of ATPα levels (5 %, p = ns) in comparison to the control group. In the comparison between cardiomyopathy groups, ATPα levels in CCC were 34 % lower than in IC myocardium (p<0. 01); and 13 % lower than those found in IDC myocardium (p<0. 05). There was no significant decrease of ATPβ in CCC when compared to control samples (9 %, p = ns). However, we observed increased expression of ATPβ in IC and IDC myocardium when compared to the control group [32 % (p<0. 001) and 10 % (p<0. 05), respectively]. In the comparison between cardiomyopathy groups, we found that CCC myocardium samples express significantly less ATPβ than IC or IDC myocardium [31 % (p<0. 001) and 17 % (p<0. 01), respectively]. We have also detected significant protein expression differences among enzymes of the creatine kinase system. The expression of CKM (Figure 2C) was decreased in samples from patients with CCC (33 %, p<0. 01), IDC (23 %, p<0. 05) and IC (24 %, p<0. 05) when compared to the control group. Of note, in the comparison between cardiomyopathy groups, the average CKM expression was most decreased among CCC patients (13 % and 12 % reduction when compared to IDC and IC, respectively), although the difference was not statistically significant. The protein expression of CKMit (Figure 2D) was decreased in samples from patients with CCC and IDC, when compared to the control group (16 % and 4 %, respectively), but the differences failed to achieve statistical significance. In contrast, samples from patients with IC showed an increased expression of CKMit when compared to the control group (13 %; p = ns). In the comparison between cardiomyopathy groups, the protein levels of CKMit were decreased in samples from patients with CCC when compared to samples from patients with IC and IDC [26 % (p<0. 01) and 13 % (p = ns), respectively]. We also analyzed the mRNA expression of the enzymes tested above, ATPα, ATPβ, CKM and CKMit. Figure 3 shows the values of relative quantification of mRNA expression of these enzymes. We found that mRNA expression of CKM was decreased in samples from patients with CCC and IDC when compared to samples from subjects without cardiomyopathy [78 % (p<0. 05) and 69 % (p<0. 05) ]. Also, the mRNA expression of CKMit was reduced in CCC samples (75 %, p<0. 05) when compared to samples from subjects without cardiomyopathy. The average expression of mRNAs for all 4 enzymes was reduced in the 3 cardiomyopathies when compared to control samples, and samples from CCC patients showed the lowest expression levels. However, due to high interindividual variation of expression within each group, most of the comparisons were not statistically significant. Interestingly, mRNA levels of ATPα and ATPβ were not increased in samples from patients with IC in comparison to samples from control group, as seen in the analysis of protein expression. In the comparison between cardiomyopathy groups, none of the enzymes analyzed showed significant differences in mRNA expression for ATPα and ATPβ. In order to evaluate whether the differential protein expression of enzymes of the creatine kinase system observed above had an impact on myocardial enzyme activity, we compared total creatine kinase activity among groups (Figure 4). The creatine kinase enzyme activity was reduced in samples from patients with CCC (59 %, p<0. 01), IDC (35 %, p<0. 01) and IC (31 %, p<0. 01) when compared to samples from the control group (Figure 4). Of interest, creatine kinase enzyme activity was significantly reduced in myocardial samples from patients with CCC, when compared to IDC and IC patients [37 % (p<0. 05) and 41 % (p<0. 05), respectively]. In this paper, we observed a reduced expression of CKM, a key enzyme in myocardial energetic metabolism, in several dilated cardiomyopathies. Most importantly, we found that CCC myocardium shows significantly reduced levels of protein expression of ATP synthase alpha subunit and total creatine kinase enzyme activity, when compared to IDC or IC. We observed that the myocardial creatine kinase system shows impaired function in patients with all forms of cardiomyopathy. The reduced myocardial protein expression of CKM, observed in all cardiomyopathy groups, was reflected in the reduced total creatine kinase activity. This has been previously described for IDC and IC [35]. The reduced protein expression of CKM was probably due to transcriptional regulation at least in the cases of CCC and IDC, since CKM mRNA expression was also significantly reduced in samples from patients of such groups, when compared to samples from individuals without cardiomyopathy. Animals genetically deficient in CKM develop myocardial hypertrophy and left ventricular dilation [36], as well as higher susceptibility to mitochondrial damage and cardiac disturbances in calcium homeostasis after ischemia and reperfusion [37]. Myocardial ATP flux through the CK system was shown to be reduced by 50 % in patients with heart failure [38]. Since the CK reaction is the prime source of the myocardial energy reserve, the deficit in ATP flux through CK may contribute to the pathogenesis of heart failure [39]. The finding that average CKM and CKMit levels from CCC samples were the lowest among all groups, and that total CK activity in CCC samples was significantly lower than that of the other cardiomyopathy groups, indicates that CCC patients may show a stronger functional impairment in the CK system than other etiologies of dilated cardiomyopathy. It is likely that the significantly reduced myocardial expression of CKMit, observed in comparison to IC, - and to a lesser degree also in comparison to control group - may have contributed to the reduction in the total CK activity observed in CCC samples. Regarding the discrepancy between the significantly reduced CKMit mRNA levels and the less prominent reduction of CKMit protein levels observed in CCC, it could be due to an increased stability of this protein. The loss of CK activity in isolated hearts has been reported to cause a decline in ATP delivery to myofibrils, leading to a blunted contractile reserve [26]. Reduced energy reserve via creatine kinase, as indicated by reduced phosphocreatine/ATP ratios, limits cardiac performance during metabolic stress conditions [40]. Significantly, CCC patients have been reported to display an impaired myocardial contractile response to dobutamine [41]. It is thus possible that this reduced contractile reserve is a consequence of the significant derangement in CK activity reported here in CCC myocardium. A correlation has been reported between decreased total CK activity and LV dysfunction [42]. However, samples from CCC patients studied here showed lower total CK activity than those of IDC or IC patients, despite the fact that LV dysfunction status was similar in CCC, IC and IDC patients. This may indicate that there are disease-specific factors that induce a stronger reduction in CK activity in CCC, when compared to non-inflammatory cardiomyopathies. While the creatine kinase system may buffer transient changes in ATP levels, the rate of oxidative ATP synthesis must be closely matched to the rate of consumption. Most myocardial ATP is generated through the mitochondrial oxidative phosphorylation (complex I-V). In our study, we also found changes in the protein expression of ATPα and ATPβ (belonging to the F1 subunit of ATP synthase complex - complex V). The finding that ATPα was only reduced in CCC myocardium, but not in IC and IDC, indicates that these patients could be at a greater impairment in cardiac ATP supply, as suggested by studies in animal models of heart failure [43]. However, in our study, patients with IC showed elevated protein levels of ATPα and ATPβ, and patients with IDC showed elevated levels of ATPβ when compared to the myocardium of subjects without cardiomyopathy. Significantly, a study showed an increase in mRNA of oxidative phosphorylation components in chronic ischemia due to severe atherosclerosis [44]. In vivo measurements in normal hearts subjected to adrenergic stress have shown that ATP production by oxidative phosphorylation increases with the demand, while ATP production by the CK system remains unchanged [39]. Authors thus suggest that the ratio of ATP production by the CK system to oxidative phosphorylation decreases upon demand; in addition, the ratio may be even lower in resting hearts from heart failure patients, due to a decreased CK flux [39]. Since a reduced CK flux may be one of the most prominent metabolic abnormalities in heart failure [39], the findings of selectively reduced CK activity - and perhaps also oxidative phosphorylation activity - may suggest that energy production in CCC myocardium can be especially restricted in situations of increased demand. Inflammation associated to the significant lymphocytic infiltrate may play an important role in multiple steps of CCC pathogenesis. Inflammatory cytokines such as IFN-gamma and TNF-alpha, abundantly produced in the inflammatory milieu of CCC heart tissue, are known to induce gene expression changes in cardiomyocytes [12], [45], [46], and may directly influence energy metabolism. It has been shown that in vitro treatment with IFN-gamma inhibited the oxidative metabolism [31], and increased the rate of ATP depletion in cardiomyocytes [32]. Additionally, studies with cultured human skeletal muscle cells demonstrated that IFN-gamma treatment could inhibit the CK activity [33]. In summary, we reported that CK activity and ATPα levels are significantly reduced in CCC myocardium when compared to IDC and IC samples. If confirmed by studies with a higher number of samples, one could hypothesize that these findings could contribute to the contractile dysfunction, loss of inotropic reserve and worse outcome of CCC when compared to cardiomyopathies of non-inflammatory etiology. In vivo analysis of CK flux rate and ATP synthesis though oxidative phosphorylation may allow further validation of the present findings.

Title: Selective Decrease of Components of the Creatine Kinase System and ATP Synthase Complex in Chronic Chagas Disease Cardiomyopathy Summary: Chronic Chagas disease cardiomyopathy (CCC) affects millions in endemic areas and is presenting in growing numbers in the USA and European countries due to migration currents. Clinical progression, length of survival and overall prognosis are significantly worse in CCC patients when compared to patients with dilated cardiomyopathy of non-inflammatory etiology. Impairment of energy metabolism seems to play a role in heart failure due to cardiomyopathies. Herein, we have analyzed energy metabolism enzymes in myocardium samples of CCC patients comparing to other non-inflammatory cardiomyopathies. We found that myocardial tissue from CCC patients displays a significant reduction of both myocardial protein levels of ATP synthase alpha and creatine kinase enzyme activity, in comparison to control heart samples, as well as idiopathic dilated cardiomyopathy and ischemic cardiomyopathy. Our results suggest that CCC myocardium displays a selective energetic deficit, which may play a role in the reduced heart function observed in such patients.

lay_plos

en

Summarize: Chris Ashton was deemed surplus to requirements by England during the autumn but the Saracens winger has not given up on his international ambitions. He started just one Test in 2014 but put his hand up for a recall with a double in the 33-10 win over Munster on Saturday, taking him to 19 tries in 22 European games. Asked about his hopes of a Six Nations call-up, Ashton (right) said: ‘I don’t expect anything. 'I’ve got to fight to get back in. Hopefully Stuart (Lancaster) will recognise my hunger to play for England is as strong as ever. That’ll never change.’ Chris Ashton - seen celebrating a try for Saracens on Saturday - says he will fight for an England recall. London Irish hooker David Paice will face a disciplinary panel after his red card in the 43-41 win over Grenoble in the Challenge Cup. Paice was adjudged to have entered a ruck with his head, but the Exiles held on for their first victory since Brian Smith left. Tom Coventry, Smith’s summer replacement, wants Clark Laidlaw, cousin of Greig, to join his coaching team. London Irish hooker David Paice (middle) looks bemused at the decision to send him off against Grenoble. The great escape is still on for James Haskell’s Wasps after the England hopeful delivered a tackling masterclass in the 23-3 victory at Harlequins on Saturday night. Despite losing their first two Pool 2 games, Wasps could secure a home quarter-final if they beat Leinster at home this Saturday. Wasps are hoping to rekindle their glory days of the last decade and Haskell, who made 28 of his side’s 199 tackles at The Stoop, wants to emulate the achievements of his predecessor, Joe Worsley, whose career was cut short by a neck injury. James Haskell produced a tackling masterclass for Wasps against Harlequins on Saturday. ‘One of my heroes in rugby is Joe Worsley,’ said Haskell. ‘He would regularly put in 30 tackles a game. He taught me to tackle, so if I can get to that level I am pleased. ‘When I re-signed and came back from New Zealand (in 2012) we nearly got relegated. My knees were gone, I think Dai (Young) thought he had signed a donkey. There was lots of change. If we can keep the squad together and keep signing players, then I think we have more to give.’ A deal for All Blacks winger Frank Halai is close to completion, while Sportsmail understands that Scotland international Tim Visser is also on the club’s radar

Summary: Chris Ashton has scored 19 tries in 22 European games. Despite that the Saracens winger doesn't believe he has much chance of getting a call-up to the England Six Nations squad. The 27-year-old was deemed to requirements by England in the summer. Elsewhere, London Irish's David Paice will face a disciplinary panel after his red card against Grenoble. Wasps could yet secure a home quarter-final if they beat Leinster at home on Saturday, despite losing their first two Pool 2 games.

cnn_dailymail

en

Summarize: By. Daily Mail Reporter. PUBLISHED:. 11:59 EST, 14 February 2014. |. UPDATED:. 14:48 EST, 14 February 2014. Whinging mass murderer Anders Breivik has threatened to go on hunger strike unless prison authorities cave in to a list of demands for better conditions - including giving him a PlayStation 3. The right-wing extremist - who killed 77 people in a bomb and gun attack on July 22, 2011 - insisted he needed a better console and video games to alleviate his 'torture'-like living conditions. It emerged today that he issued a typed list of 12 demands to authorities at the high-security unit in Skien in southeast Norway where he serving out a 21-year sentence. Discomfort: Anders Behring Breivik, pictured clutching the assault rifle used during a rampage which killed 77 people in 2011, has complained that prison authorities are treating him 'worse than an animal' The demands - outlined in a letter sent to the the French national newsagency AFP today - included better conditions for his daily walk and the right to communicate more freely with the outside world, which he argues are in line with European rights legislation. He also demanded that his PlayStation 2 games console be upgraded to a Playstation 3 'with access to more adult games that I get to choose myself'. Held apart from other prisoners since 2011 for security reasons, Breivik argued in the letter sent in November, last year, that he has the right to a wider'selection of activities' than other inmates to compensate for his strict isolation. List of demands: Breivik, pictured walking among the bodies of the youngsters he killed in cold blood on Utoeya island on July 22, 2011, is demanding better conditions in the prison where he is serving a 21-year jail term. Breivik also wants his standard weekly allowance of 300 kroner ($49, 36 euros) to be doubled, particularly to cover his postal charges from written correspondence. Other demands include an end to daily physical searches, and access to a PC rather than to a 'worthless typewriter with technology dating back to 1873'. In the letter dated January 29 he said that since there has not been any real improvement in his prison conditions, a hunger strike would be 'one of the only' options at his disposal. Slaughter: A wounded woman is brought ashore opposite Utaoya island after being rescued during Breivik's killing rampage in July 2011. Mourning: Thousands of floral tributes were laid at the Oslo Cathedral, Norway, following Breivik's killing spree. 'The hunger strike won't end until the Minister of Justice (Anders) Anundsen and the head of the KDI (the Norwegian Correctional Services) stop treating me worse than an animal,' he said, adding that he would'soon' make public the starting date of his protest action. On July 22, 2011, Breivik killed eight people in a bomb attack outside a government building in Oslo and later killed a further 69, most of them teenagers, when he opened fire at a Labour Youth camp on the island of Utoeya

Summary: Breivik is serving 21 years in prison for murdering 77 people in 2011. Killer complains the prison is treating him 'worse than an animal' He sent a list of 12 demands to improve conditions to jail authorities. Details of requirements sent to officials in November emerged today. 35-year-old wants access to more adult games for his computer console. Breivik wants his weekly allowance of 300 kroner ($49, €36) to be doubled.

cnn_dailymail

en

Summarize: By. Tim Shipman. and Ian Birrell. Vladmir Putin put the world on course for a new Cold War yesterday, annexing Crimea and telling the West to get out of Russia’s back yard. As Russia was accused of war crimes after the death of a Ukrainian soldier, Putin defiantly told a joint session of the Russian parliament that he would not accept Western influence ‘next to our home or on our historic territories’. And in a stark warning that Ukraine risks dismemberment if it seeks to join the EU or Nato, he said he would not tolerate Western countries ‘behaving as the master of the house outside our fence’. Scroll down for video. Russian President Vladimir Putin signs a treaty for Crimea to join Russia. Russian President Vladimir Putin (second right), Crimea's Prime Minister Sergei Aksyonov (front left), Crimean parliamentary speaker Vladimir Konstantinov (back left) and Sevastopol Mayor Alexei Chaliy shake hands after a signing ceremony at the Kremlin in Moscow. An emotional Putin signed a document. reclaiming Crimea and declared: ‘In the hearts and minds of people,. Crimea has always been and remains an inseparable part of Russia.’ The. speech sent shockwaves through the international community and appeared. to have upended two decades of diplomacy since the fall of the Soviet. Union designed to imbed Russia as a member of the international. community. It came as a. Ukrainian officer and a member of a local Crimean self-defence brigade. were shot dead when pro-Russian militia stormed a military outpost in. the Crimean capital of Simferopol. Adoring public: Putin was last night riding the crest of an adulatory wave after righting what many Russians see as an historical wrong and reintegrating Crimea and the Black Sea fleet headquarters of Sevastopol. Nationalistic fervour: Dozens of Russian flags flutter in the breeze below a typically overcast Moscow sky. Ukrainian. Prime Minister Arseniy Yatsenyuk said the storming of the military. facility showed that the dispute ‘has gone from the political stage to. the military by the fault of the Russians’. He. added: ‘Today, Russian soldiers began shooting at Ukrainian servicemen. and this is a war crime without any statute of limitations.’ David. Cameron yesterday condemned the decision to annex Crimea ‘on the basis. of a sham referendum held at the barrel of a Russian gun’. On the crest of a wave: Russian President Vladimir Putin stands next to Crimean Premier Sergey Aksionov as he addresses a rally in Red Square, Moscow, celebrating Crimea's decision to join with Russia. Cult of personality: Russians hold flags adorned with Putin's face and a slogan reading 'We are together!' Mr Aksionov, the new Crimean premier, raises his fist as he shares the stage with Speaker of the Supreme Council of Crimea Vladimir Konstantinov, left, and head of Sevastopol city administration Alexei Chaliy, right. He. said: ‘The steps taken by President Putin today to attempt to annex. Crimea to Russia are in flagrant breach of international law and send a. chilling message across the continent of Europe. ‘President Putin should be in no doubt that Russia will face more serious consequences.’ The. Prime Minister will call on fellow EU leaders to bring in a new round. of tougher economic sanctions, plus an arms embargo, when they meet at a. Brussels summit tomorrow. Branding. the crisis ‘the most serious test of European security in the 21st. century’, Foreign Secretary William Hague made clear that two decades of. reaching out to Russia were at an end and that relations were likely to. dive. Foreign Secretary. William Hague said Britain had suspended all military co-operation with. Moscow. Suggesting Russia should be kicked out of the G8 group of. nations, he said: ‘We should be ready to contemplate a new state of. relations between Russia and the West in the coming years that is. different from the last 20 years.’ Last. night, Russian politicians and media were telling Putin to grab back. oil and gas-rich Kazakhstan and authoritarian Belarus as well as more. slices of a battered Ukraine. Senior politician Sergei Mironov hailed ‘the great day when the gathering of Russian lands began’. Adolf Hitler made a spookily similar speech to Putin when he seized the Sudetenland, making similar references to common themes. The uncompromising autocrat got to his feet and made a bombastic speech as he seized a part of a neighbouring state dominated by his own countrymen. It came in the midst of a referendum condemned by other powers as a sham, with voters intimidated by the autocrat’s looming tanks. The move was greeted with delirious pleasure by his own citizens but seen as a portent of international strife by other Western countries. The despot in question is not Vladimir Putin but Adolf Hitler. The occasion was not the annexation of Crimea but the Nazi seizure of the Sudetenland, the German-speaking part of Czechoslovakia. Putin, who spoke to the Russian parliament yesterday, appears to have stolen many of the themes of his speech from the Fuhrer’s address to the Reichstag as he seized the Sudetenland in October 1938. Putin said Crimea had always been Russian: ‘In people’s hearts and minds, Crimea has always been an integral part of Russia.’ Hitler said of the Sudetenland: ‘The Sudeten German population was and is a German.’ Putin insisted he would fight for Russians everywhere. Hitler’s imagery was also emotive. ‘More than 1,000,000 people of German blood had in the years 1919-1920 to leave their homeland.’ Putin criticised Western nations, and Hitler complained that his efforts to reach out to the West were rebuffed.Putin said Russia would retaliate against Western sanctions. Hitler said: ‘As a National Socialist, I am accustomed to strike back at an attacker.’ Putin claimed he would not invade other parts of Eastern Ukraine if Russia was allowed to exercise influence: ‘Don’t believe those who try to frighten you with Russia and who scream that other (Ukrainian) regions will follow after Crimea. We do not want a partition of Ukraine.’ Hitler also claimed that his ambitions would stop at the Sudetenland. ‘I have declared that the frontier between France and Germany is a final one. Germany has no interests in the West.’ Five months later, he invaded the rest of Czechoslovakia; 14 months after that, he invaded France

Summary: Putin signs bill to annexe Crimea and pull territory back into Russia. Believes the 'hearts and minds' of Crimea want to be in Russia. Warns the West not to interfere in his annexing of Crimea. The annexing has been largely condemned by the international community. Hours after speech, one Ukrainian officer was shot dead in Crimea. Unmarked men, thought to be Russian, stormed a Ukrainian compound. Fired shots, arrested and detained many, leading to Ukrainian death. Ukraine has referred to this incidence as a 'war crime' Ukraine ordered its troops to fight back in self-defence from now on. Interfax news agency reported a pro-Russian militia also died in gunfire.

cnn_dailymail

en

Summarize: BACKGROUND OF THE INVENTION 1. Field of the Invention The invention pertains to buoyant fishing bobbers molded of synthetic plastic material and wherein a fishing line latch or retainer is homogeneously hinged to the basic bobber structure. 2. Description of the Related Art While two-piece fishing bobbers are known wherein the basic body of the bobber is formed of two interconnectable portions, as shown in U.S. Pat. Nos. 2,803,083 and 3,141,256, such prior art devices normally produced a separable body chamber for the purpose of storage of the line, hooks and sinkers. Also, while it is known to incorporate homogeneous integral line attachment components with fishing line accessories, such as sinkers, as shown in U.S. Pat. Nos. 4,964,236 and 5,241,776, the utilization of integral fish line holding structure with a bobber is not common. Prior art fishing bobbers molded of synthetic plastic material are of such complexity as to be relatively expensive, and bobbers utilizing pivoting fish line retainers do not incorporate the low cost advantage of pivotally hinging the fish line latch to the bobber structure by a homogeneous hinge portion or element wherein the latch and the associated bobber structure are formed as an integral one-piece unit. OBJECTS OF THE INVENTION It is an object of the invention to provide a low cost moldable synthetic plastic fishing bobber having a flotation or buoyancy chamber formed of two portions which may be readily separated to permit ballast to be located within the chamber to adjust buoyancy sensitivity. Another object of the invention is to provide a low cost moldable fishing bobber having a lower stem upon which a fishing line latch is homogeneously molded pivotal about a homogeneous hinge between open and closed positions to selectively retain or release the fishing line. Yet another object of the invention is to provide a fishing bobber molded of a synthetic plastic material which is of very low cost, effective and efficient in operation, and capable of effectively adjustably gripping a fishing line, and releasing the same, requiring no special skills on the part of the fisherman. SUMMARY OF THE INVENTION The fishing bobber of the invention is molded of a synthetic thermoplastic material whereby the bobber may be economically manufactured without requiring secondary operations. The bobber consists of an upper part which is releasably connected to a lower part at a fluid tight connection. The connectable portions, together, define a hollow body or cavity which produces the bobber buoyancy. By temporarily separating the upper and lower parts, ballast, such as water or other matter, may be placed within the cavity to add weight to the bobber causing it to float low in the water and increasing the sensitivity of bobber to fish feeding on the fishing line depending from the bobber. The lower part of the bobber includes a homogeneous elongated stem having a lower end and a latch is pivotally defined on the stem by a homogeneous hinge portion formed of the material of the stem and latch. The upper end of the latch associates with a lock formed on the stem whereby the latch may be held parallel to the length of the stem with a fishing line pinched between the latch and the stem permitting the line to be adjustably affixed to the bobber. The flexible thermoplastic material of which the bobber is formed permits the stem to be laterally deformed or bent which will release the latch from the stem lock permitting the latch free end to pivot away from the stem to free the line. The construction of a fishing bobber in accord with the invention permits the bobber to be constructed at a low cost, is dependable in operation, and may be used by fisherman of average skill to effectively attach and release a fishing line to a bobber. BRIEF DESCRIPTION OF THE DRAWINGS The aforementioned objects and advantages of the invention will be appreciated from the following description and accompanying drawings wherein: FIG. 1 is an elevational view of a fishing bobber in accord with the invention, the upper and lower parts thereof being connected, and the latch being shown in the closed position, FIG. 2 is an elevational view of the invention illustrating the upper and lower parts separated, and the latch hinged to the open position, FIG. 3 is a detail enlarged elevational view of the lower end of the bobber stem illustrating the fishing line latch in the closed position retaining a fishing line to the bobber, FIG. 4 is an enlarged detail elevational view of the lower end of the stem upon being laterally deformed to release the latch lock, FIG. 5 is an elevational enlarged detail view of the lower end of the stem illustrating the latch in the open position, FIG. 6 is an enlarged detail elevational sectional view taken through the connecting joint of the bobber upper and lower parts, and FIG. 7 is an enlarged plan sectional view of the stem taken along Section 7--7 of FIG. 3. DESCRIPTION OF THE PREFERRED EMBODIMENT A typical general configuration of a fishing bobber utilizing the inventive concepts is shown in FIG. 1 wherein the bobber is generally indicated by reference numeral 10. The bobber 10 includes an upper body part 12 and a lower body part 14 which is releasably interconnected in a watertight manner to the body part 12 as later explained. The upper part 12 includes a bell 16 having an elongated flag 18 extending therefrom, and the lower edge 20 of the bell 16 is of a configuration as will be appreciated from FIG. 6. The lower body part 14 is of a dish configuration at 22 and includes a downwardly extending elongated stem 24. The upper edge 26 of the dish 22 is shown in FIG. 6 and is complementary in configuration to the edge 20. Preferably, the bobber parts are molded of a synthetic thermoplastic material of an attractive low cost characteristic having flexing ability, such as a polypropylene. The material may be colored, or clear, as desired. The flag 18 is integral and homogeneous with the bell 16, while the stem 24 is integral and homogeneous with the dish 22. With modern injection molding equipment, the parts 12 and 14 may be simultaneously formed and automatically ejected from the molding die at the end of formation. The stem 24 includes a free cantilevered lower end 28 which may be of a convex configuration as shown in full lines in the drawing. A latch 30 homogeneously formed of the material of the stem 24 includes a hinge portion 32 homogeneously defined on the stem lower end 28 and the hinge 32 is formed of the material of the stem and is integral both with the stem lower end and the lower region of the latch 30. The homogeneous hinge 32 permits the latch 30 to be pivoted between the closed position shown in FIG. 3 and the open position shown in FIG. 5. A bulbous projection 34 is defined on the latch 30 adjacent the hinge 32 and this projection is complementary to a recess 36 defined in the lower region of the stem 24 for receiving the fishing line as later described. As will be appreciated from the drawings, the stem 24 includes a reduced semi-circular portion 38 adjacent the latch 30, and the stem also includes a lip 40 which extends downwardly toward the stem lower end 28. A tongue 42 is defined on the upper free end of the latch 30, and the upper end of the latch also includes an inclined ramp 44 which, when the latch 30 is pivoted from the position of FIG. 5 to the position of FIG. 3, engages the lip 40 slightly bending stem portion 38 and depressing the latch 30 downwardly so that the tongue 42 will be received under the stem lip 40 and in this manner, the lip 40 will retain the latch 30 in the closed position shown in FIG. 3. The fish line 46 may be inserted between the latch 30 and the stem portion 38 when the latch is in the open position shown in FIG. 5. The fish line 46 is preferably located within the recess 36 so that when the latch 30 is pivoted to the closed position of FIG. 3, the projection 34 will tightly frictionally engage the fish line 46 so that the parts are related as shown in FIG. 3. As the projection 34 is located adjacent the hinge 32, and as the projection 34 and recess 36 are complementary in configuration, the fish line 46 will be tightly squeezed between the projection and recess establishing a firm frictional connection between the fishing bobber and fishing line. The interconnection between the upper body 12 and the lower body part 14 is best illustrated in FIG. 6. As shown, the edge 20 of the part 12 includes an annular concave recess 48, and the upper edge 26 of the dish 22 includes a rounded convex head 50 complementary in configuration to the recess 48. The flexibility of the material of the parts 12 and 14 permits the head 50 to be popped out of the recess 48 to separate the parts 12 and 14, and in this manner, when the parts are separated, ballast in the form of water, lead pellets, or the like, may be placed within the cavity defined by the parts 12 and 14 to add weight to the bobber and vary the buoyancy and sensitivity of the bobber during fishing. In use, assuming the latch 30 to be in its latched position shown in FIG. 3, the user need only slightly deform the stem 24 as shown in FIG. 4 to permit the latch tongue 42 to ride under the stem lip 40 permitting the latch 30 to pivot to its open position as shown in FIG. 5. At this time, the fishing line 46 may be easily removed from the recess 36 and the position of the bobber on the line may be adjusted or the fish line removed. When the latch 30 is pivoted to the closed position shown in FIG. 3, the ramp 44 initially engages the lip 40 which causes a slight deformation of the stem 24 and the fully latched position is shown in FIG. 3. The natural resiliency of the material of the stem 24 permits the stem to be readily deformed during the opening and closing operations of the latch, and by forming the bobber parts of polypropylene or similar material, the hinge 32 is capable of thousands of repetitions without breakage. If it is desired to reduce the force necessary to pivot the hinge 30 between open and closed positions, the wall thickness of the stem adjacent the hinge 32 may be reduced by terminating the end of the stem along a plane such as shown at 52 in FIG. 3 in dotted lines. In such a situation, the amount of material in the hinge 32 is minimized permitting the hinging of the latch to be easily accomplished. It is appreciated that various modifications to the inventive concepts may be apparent to those skilled in the art without departing from the spirit and scope of the invention.

Summary: A fishing bobber molded of a synthetic plastic material wherein a hollow flotation chamber includes separable upper and lower snap together parts whereby ballast may be located within the chamber to adjust flotation sensitivity, and an elongated downwardly extending stem includes a latch having a hinge homogeneously formed of the stem material wherein the latch is pivotally positionable between a closed position for gripping the fishing line, and an open position for releasing the line. Manual lateral deformation of the stem unlocks the latch from its closed position.

big_patent

en

Summarize: Background Recovery auditing has been used in various industries, including health care, to identify and collect overpayments for about 40 years. Private insurance companies, managed care plans, and employee group health plans contract with recovery auditors to review payments made. Typically, recovery auditing contractors are paid a contingency fee based on a percentage of the overpayments collected. Fees vary depending on such factors as the types of overpayment involved and the degree of difficulty associated with identifying and collecting them. Use of Contractors in the Operation of the Medicare Program Contractors play an essential role in the Medicare program. Since the program’s inception in 1965, Medicare claims administration contractors, then known as fiscal intermediaries and carriers, have conducted its claims administration activities. In addition, CMS also uses other contractors to conduct Medicare functions, such as to investigate instances of potential fraud and develop cases for referral to law enforcement and to answer beneficiary inquiries through the 1-800-Medicare help line. At present, CMS is in the midst of the largest transition of its claims administration contracts since the program was established. The MMA required CMS to use competitive procedures to select new entities called Medicare Administrative Contractors (MACs) to conduct claims administration activities that had been conducted by fiscal intermediaries and carriers. Through February 2005, CMS contracted with approximately 51 fiscal intermediaries and carriers that processed and paid claims, conducted automated pre-payment and limited post-payment review of claims, handled the first level of provider appeals of denied claims, enrolled providers in Medicare, and audited providers’ cost reports. To address improper billing, these Medicare claims administration contractors also performed trend analysis of provider billing patterns, developed strategies to address improper billing through systems edits or provider education and claims review, helped implement CMS-issued national coverage determinations (NCD), and developed local coverage determinations (LCD). By the end of the transition from fiscal intermediaries and carriers to MACs, CMS will have transferred all of these tasks to 15 MACs that will handle Part A claims and Part B claims with the exception of durable medical equipment (DME) claims, which will be processed by four specialized DME MACs. As of September 2009, CMS made an initial award decision on all the MAC contracts and has implemented 13. Because the transition is not completed, the current Medicare contracting environment includes fiscal intermediaries, carriers, and MACs, any one of which we refer to as Medicare claims administration contractors for this report. Claims Review in Medicare Medicare claims administration contractors review Medicare claims both before and after payment using similar automated and complex processes. CMS’s use of recovery auditing in the RAC demonstration project augmented existing Medicare claims administration contractor pre- and post-payment claims review efforts. While Medicare claims administration contractors have the authority to review claims they initially paid, this is only one of the many functions they perform. Further, because the Medicare claims administration contractors receive more than 1.2 billion claims per year (the equivalent of 4.5 million claims per work day), it is impractical, according to CMS, for these contractors to manually review more than a small fraction of claims—either before or after payment. Recovery audit contractors, in contrast, focus exclusively on post-payment claims review. Medicare claims administration contractors and the RACs generally use the same processes to review claims: Automated reviews use systems edits to check claims for evidence of improper coding or other mistakes. Medicare claims administration contractors may use automated reviews before payment to deny claims, or to flag claims that require additional non-automated review before payment. RACs use automated reviews after payment to analyze paid claims and identify those that were or could have been paid improperly. Complex reviews rely on licensed medical professionals to manually examine a claim and any related documentation, including paper files, to determine whether the service was covered and was reasonable and necessary. Complex reviews conducted by a Medicare claims administration contractor or a RAC involve an examination of the medical records associated with a service, which the provider submits for review. RAC Responsibilities in the Demonstration Project CMS implemented the RAC demonstration project to test whether recovery auditing would effectively identify additional improper payments that could be recouped. In March 2005, CMS selected three RAC contractors to conduct claims reviews in the three states with the highest per-capita Medicare utilization rates—California, Florida, and New York. In July 2007, CMS expanded the demonstration project to three additional states—Arizona, Massachusetts, and South Carolina. The demonstration project ended in March 2008. CMS initially provided the RACs with 4 years of claims data in their jurisdictions, followed by an additional 3 months of claims each quarter for the rest of the demonstration project. CMS gave the demonstration RACs a total of 1.2 billion claims that they could review. To prevent the RACs from auditing those claims that previously underwent complex review by a Medicare claims administration contractor or other contractor, CMS established a data warehouse that contained information on which claims were unavailable for RAC review. During the demonstration project, the RACs were required to use automated and complex review processes using the same Medicare policies and regulations as CMS’s Medicare claims administration contractors to identify improper payments. The RACs used their own software to analyze paid claims and identify those that were or could have been paid improperly. For example, claims indicating duplicate payments could be identified by automated analysis alone. In other cases, the RACs identified claims likely to contain errors and conducted complex reviews. (See fig. 1 for a depiction of the claims review process.) In these cases, the RACs requested that providers submit the associated medical records for review. If the RAC found an improper payment, it notified the provider and the Medicare claims administration contractor responsible for recouping the overpayments or repaying an underpayments. Providers could appeal RAC determinations through the established Medicare appeals process, which included a first-level review conducted by the Medicare claims administration contractors. Two years into the demonstration project, CMS initiated a series of vulnerability calls, conference calls between the RACs and the Medicare claims administration contractors. These calls enabled the RACs to provide information about the vulnerabilities they identified that resulted in improper payments and to highlight situations where corrective action might be needed. Although a CMS official told us it was not required, the Medicare claims administration contractors could consider RAC-identified vulnerabilities when developing their strategies to reduce improper payments. If a Medicare claims administration contractor determined that a RAC-identified vulnerability was widespread in its region, it could choose to take several corrective actions. A Medicare claims administration contractor could: (1) conduct provider outreach and education, (2) develop or revise local coverage determinations to clarify what services were reasonable and necessary in that jurisdiction, and (3) initiate additional service-specific prepayment edits in its local claims processing system. In addition, CMS could initiate a nationwide corrective action, such as implementing a national system edit, reissue instructions for coding a claim, or develop a national coverage determination. CMS also could provide outreach and education on critical issues to providers directly through its Special Open Door Forums teleconferences, and presentations at national meetings. In its June 2008 evaluation report, CMS stated that the demonstration project corrected $1.02 billion in improper payments from the three claim RACs—$980.0 million in overpayments and $37.8 million in underpayments—as of March 27, 2008, and returned $693.6 million to the Medicare Trust Funds. Eighty-five percent of the overpayments collected were for services detailed on inpatient hospital claims. Common types of improper payments were for claims determined to be: coded incorrectly, lacking sufficient documentation, or medically unnecessary. However, the RACs collected the majority of these improper payments in the last quarter of the demonstration project, and many provider appeals had not been decided or even filed by the end of the demonstration project. The final outcome of the appeals process, which can take more than two years, could decrease the savings attributed to the demonstration project. CMS’s report also discussed several changes the agency made prior to the start of the RAC national program. (See app. I.) RAC Responsibilities in the National Program In 2008, following the mandate to create a national program, CMS made initial awards of contingency-fee contracts to four RACs, each with responsibility for reviewing claims in one of four geographic regions. CMS launched the RAC national program in two stages with outreach activities beginning in 24 states on March 1, 2009, and the remaining states starting in August 2009 or later. (See fig. 2.) RAC claim reviews in the national program involve the same processes of automated and complex review of claims as during the demonstration project, and the Medicare claims administration contractors are responsible for recoupments, claims adjustments, and provider outreach and education. The four regional RACs also are required to conduct outreach to providers about the purpose of the RAC program, assist CMS with the development of an improper payment prevention plan, and support the agency regarding any overpayments appealed by providers. The RACs are expected to conduct outreach to providers in each state in coordination with CMS and include the appropriate Medicare claims administration contractor in each state in its region. In addition, RACs are required to compare the claims proposed for review with the claims in the data warehouse to ensure that a Medicare claims administration contractor or other contractor had not previously audited the claims or that RAC activities would not interfere with potential fraud investigations. From March 2009 through June 2009, the RACs’ activities included accessing claims data from CMS and convening meetings with the providers in the states in their regions to explain the RAC program. In June 2009, CMS announced a gradual implementation of claims review activities. CMS permitted RACs to begin automated reviews as of June 2009. RACs will be permitted to conduct complex reviews to assess medical necessity of DME claims in fiscal year 2010 and complex review of other claims for medical necessity in calendar year 2010. (See fig. 3 for a timeline for the RAC program.) CMS Did Not Establish an Adequate Process to Address RAC-Identified Vulnerabilities That Led to Improper Payments; Corrective Actions Were Limited CMS did not establish an adequate process during the demonstration project or in planning for the national program to ensure prompt resolution of the RAC-identified improper payment vulnerabilities. Although the agency’s goal was for the RACs to provide information to CMS and Medicare claims administration contractors that could help prevent future improper payments, CMS did not implement corrective actions for 60 percent of the most significant vulnerabilities identified during the RAC demonstration project. CMS Did Not Establish an Adequate Process to Address RAC-Identified Vulnerabilities to Reduce Improper Payments While CMS stated in its fiscal year 2006 status report on the RAC demonstration project that the agency intended to draft a corrective action plan to prevent future improper payments based on the findings identified by the RACs, it did not do so. CMS developed the IPPP—a list of the most significant vulnerabilities that led to improper payments and corrective actions taken to address them—but this document did not include the essential elements of a corrective action plan. The IPPP listed the 58 most significant RAC-identified vulnerabilities—generally those that resulted in overpayment collections of $1 million or more—and whether any corrective actions were taken to address them. Improper payments for medically unnecessary services and duplicate claims are examples of types of RAC-identified vulnerabilities listed in the IPPP. For each vulnerability, the IPPP listed the provider type, improper payment amount, status, and comments. If any action were taken by CMS or its Medicare claims administration contractors, it would be noted in the IPPP. For the RAC national program, CMS has yet to assign responsibility to personnel for implementing corrective actions to address RAC-identified vulnerabilities or to develop steps to assess the effectiveness of actions taken. Based on criteria outlined in our Standards for Internal Control in the Federal Government and criteria that CMS developed for a corrective action process, we found the following limitations in CMS’s resolution process: CMS lacked a process to evaluate RAC findings promptly. CMS did not begin to evaluate the most significant vulnerabilities that resulted in improper payments until almost 2 years after the program began. Agency officials told us they did not anticipate that the RACs would identify such a high volume of improper payments and did not have systems in place to collect data at the beginning of the demonstration project. CMS’s fiscal year 2006 status report on the RAC demonstration project stated that CMS would draft a proposed RAC Corrective Action Plan to prevent future improper payments by January 2007. However, CMS did not create the IPPP—the spreadsheet to track significant vulnerabilities identified during the demonstration project—until November 2008, 8 months after the demonstration project ended. CMS lacked a process to determine appropriate responses to RAC findings. CMS did not assign responsibility for taking corrective action on the vulnerabilities listed in the IPPP to either the agency itself, its Medicare claims administration contractors, or a combination of both. According to CMS officials, the agency only takes corrective action for vulnerabilities with national implications, and leaves it up to the Medicare claims administration contractors to decide whether to take action for vulnerabilities with local implications. However, the IPPP did not specify what type of action was required on the part of CMS or the Medicare claims administration contractors. For example, for inpatient services that did not meet the stated inpatient care criteria, the IPPP neither specified what type of corrective action would be needed to prevent future improper payments nor whether CMS or its Medicare claims administration contractors were responsible for taking action. Accordingly, neither Medicare claims administration contractors nor CMS have taken corrective action to address payment errors related to this inpatient service vulnerability. Similarly, we reviewed the instructions CMS provided to the Medicare claims administration contractors during the demonstration project and found that CMS did not provide specific guidance to the Medicare claims administration contractors for incorporating RAC findings into local corrective action plans. Instead, CMS allowed its Medicare claims administration contractors to independently determine when to take action and what actions, if any, were needed to address RAC findings. The lack of documented assigned responsibilities—as prescribed in our internal control standards— impeded CMS’s efforts to promptly resolve the vulnerabilities identified by the RACs during the demonstration project. CMS lacked a process to implement corrective actions promptly. The IPPP, which was not created until 8 months after the end of the demonstration project, lacked a time frame based on established criteria for when CMS or its Medicare claims administration contractors should take action. CMS officials told us that although they conducted some informal follow-up, neither the agency nor its Medicare claims administration contractors have implemented any corrective actions to address RAC findings since the fall of 2008. CMS officials noted that the agency does not plan to take any further action until the appeals from the demonstration project are finalized. Because CMS has not developed a time frame for taking action based on established criteria and is currently unable to track all pending first-level appeals of RAC determinations, it is uncertain when or if the agency would take any further action on the remaining vulnerabilities. Although educating providers promptly on how to correct billing errors reduces the risk of improper payments, provider associations also told us they and their members had not received training on the majority of the vulnerabilities identified by the RACs during the demonstration project. For example, one national provider association said that it was not aware of any educational efforts related to the RAC program findings on vulnerabilities either during or after the demonstration project. Another noted that in addition to provider education, systems edits should be used when possible to prevent the initial improper payments. CMS continues to lack an adequate process for implementing corrective actions during the RAC national program. Although CMS has made public statements that preventing future improper payments is the RAC program’s mission, the agency has yet to assign responsibility to personnel for implementing corrective actions to address RAC-identified vulnerabilities or to develop steps to assess the effectiveness of corrective actions taken. While CMS’s Office of Financial Management (OFM) established a corrective action team for the RAC national program that will compile, review, and categorize RAC-identified vulnerabilities and discuss corrective action recommendations, the team does not have the organizational authority to implement the corrective actions necessary to reduce future improper payments. Rather, the team can only forward the issues and their recommendations to other leadership groups comprised of senior officials from different components within CMS that have the authority to take corrective actions. For example, if the decision is made to address a vulnerability by developing a NCD, the responsibility to prioritize the development of NCDs and expertise to develop them is not within OFM, but rather within the Office of Clinical Standards and Quality. The different components can choose whether to address the identified vulnerabilities that could lead to improper payments. Further, CMS’s corrective action process does not include steps to assess the effectiveness of any actions taken to reduce improper payments on RAC-identified vulnerabilities. Strong internal controls include ongoing monitoring of corrective actions, evaluating their effectiveness, and modifying them as necessary. CMS officials in OFM said their corrective action team would monitor actions taken by other agency components. However, the corrective action process does not include any steps to either assess the effectiveness of the corrective actions taken or adjust them as necessary based on the results of the assessment. Until CMS designates key personnel with accountability for ensuring corrective actions are implemented and establishes a process to ensure these actions are effective, the agency remains at risk for making improper payments on vulnerabilities previously identified by RACs. CMS’s Corrective Actions Did Not Address Most of the RAC-Identified Vulnerabilities That Led to Improper Payments The lack of accountability and adequate processes for ensuring corrective actions are taken have resulted in most of the RAC-identified vulnerabilities that led to improper payments going unaddressed. CMS implemented corrective actions for 23 of the 58 vulnerabilities (40 percent) listed in the IPPP. (See fig. 4.) This left 35 of the 58 vulnerabilities identified during the demonstration project (60 percent) unaddressed, representing millions of dollars in potential overpayments. CMS stated in its June 2008 demonstration evaluation report that overpayments were identified for 18 specific medical services totaling $378 million. Our analysis of the status of the vulnerabilities related to these overpayments in the IPPP indicates that corrective actions had not been implemented by CMS or the Medicare claims administration contractors for vulnerabilities representing $231 million (61 percent) of the $378 million in overpayments for these services. More than 90 percent of the $231 million in vulnerabilities that were not addressed were for inpatient hospital claims alone. The corrective actions taken to address 23 of the 58 vulnerabilities (40 percent) included: 7 system edits (12 percent), 6 provider education activities (10 percent), and 10 clarifications of guidance and issuance of new regulations (17 percent). Six of the 23 corrective actions taken included local actions implemented by the Medicare claims administration contractors and other contractors, but according to the IPPP, CMS also implemented national corrective actions for the same vulnerabilities. CMS did not implement corrective actions for 35 of the 58 vulnerabilities (60 percent) listed in the IPPP. Of these 35 vulnerabilities, CMS did not list a reason on the IPPP for 28 of them (48 percent). CMS officials told us that they were unable to develop specific corrective actions on the other seven (12 percent) because they either lacked adequate information to address the problem or decided it was not cost-effective to do so. CMS officials told us the agency was unable to develop corrective actions for 7 vulnerabilities because the agency did not provide sufficient guidance to the RACs on how to categorize these vulnerabilities. As a result, the RACs combined several billing codes into single categories, which presented a challenge for identifying corrective actions, according to CMS officials. For example, RACs denied millions of dollars in inpatient hospital claims not meeting the requirements for inpatient admission. However, CMS officials told us they were unable to develop corrective actions on this and six other vulnerabilities because they either lacked adequate information on specific services involved or decided it was not cost effective to address each specific billing code. Further, the agency reported that it did not have sufficient time to analyze the information on one of these types of vulnerabilities prior to the end of the demonstration project. CMS noted several actions it took to improve the quality of its information on improper payment vulnerabilities that might be identified through the national RAC program. According to CMS officials, the agency has enhanced the data warehouse to provide additional information by establishing 20 to 30 different types of categories for use in the national program. In addition, CMS officials said they will not rely on each RAC to report its findings; instead, the agency will use the information from the data warehouse for data analysis and reports. CMS officials told us they had no plans to take further action on RAC- identified improper payment vulnerabilities that have appeals outstanding from the demonstration project until the results from these appeals are known. According to the agency, information from these appeals may help the agency determine what corrective actions are appropriate. CMS and Medicare claims administration contractors reported that the following factors also hindered their progress in implementing corrective actions: Competing priorities in implementing system edits—According to CMS officials, national systems edits to address RAC findings competed with other computer system changes, such as Medicare fee schedule updates. National edits require collaboration among various CMS components and senior executives to determine the viability of each edit and its priority level and can take up to 7 months to be implemented. The decision to implement system edits at the local level is usually up to the local Medicare claims administration contractor. A Medicare claims administration contractor can decide not to implement a local edit if it does not consider that particular vulnerability a priority in its strategy to reduce improper payments or if it anticipates that the edit would result in a high level of appeals. CMS officials also told us that the availability of resources, including staff hours, played a role in prioritizing the implementation of national and local edits. Due to the limited resources available and the agency’s competing priorities, RAC-related system edits from the three state demonstration project were not a high priority according to CMS. Significant workload increase in processing claim readjustments and appeals—CMS officials and one of the Medicare claims administration contractors’ staff we interviewed told us that the increase in workload from claim adjustments and appeals from RAC findings during the demonstration project strained the Medicare claims administration contractors’ capacity to institute corrective actions. Medicare claims administration contractors made adjustments for claims in which the RACs had identified either overpayments or underpayments. However, during the demonstration project, the Medicare claims administration contractors processed hundreds of thousands of RAC claim adjustments—some manually—which created significant additional workload. In addition, both of the Medicare claims administration contractors that we interviewed that worked with the RACs during the demonstration project reported significant increases in appeals workload due to RAC activities, especially Part A appeals. One Medicare claims administration contractor stated that in fiscal year 2008, 99 percent of its Part A appeal workload arose from RAC claims, while another claims administration contractor reported having twice as many Part A appeals as it did prior to the demonstration project. Transition of Medicare claims administration functions to MACs— The transfer of claims administration responsibilities to MACs further contributed to CMS’s inability to implement corrective actions. CMS consolidated numerous fiscal intermediary and carrier jurisdictions into the new MAC jurisdictions. The MACs are responsible for consolidating the different coverage policies and systems edits they inherited from the previous contractors into one consistent set of edits and coverage policies for the new jurisdictions. As a result, CMS told us that some Medicare claims administration contractors did not act upon RAC-identified vulnerabilities that led to improper payments during the demonstration project. Further, CMS officials said that in part they did not implement corrective actions due to the lack of continuity when some of the Medicare claims administration contractors were not awarded MAC contracts, which prevented the agency from continuing discussions with contractor staff familiar with the RAC program. Our prior work has shown that CMS has allowed known vulnerabilities that contribute to or result in improper payments to remain unresolved for years. In fact, the RACs focused on some specific types of claims because both we and the HHS Office of the Inspector General identified them in the past. Moreover, CMS officials and one of the RACs noted that many of these vulnerabilities were known to CMS before the demonstration project due to medical record reviews and the agency’s error reports. In its 2006- 2009 Strategic Action Plan, CMS reported that it planned to effectively oversee its providers and aggressively deliver provider education and outreach and that this oversight would include ways to prevent overpayments and improper payments. In addition, CMS reported that it was also expanding the use of electronic data to more efficiently detect improper payments and program vulnerabilities. However, we have reported recently that continuing weaknesses in CMS’s process still exist, and therefore Medicare continues to be at risk for improper payments. CMS Is Taking Action to Resolve RAC and Medicare Claims Administration Contractor Coordination Issues CMS used lessons learned from the RAC demonstration project to take actions to resolve RAC and Medicare claims administration contractor coordination issues for the RAC national program. Specifically, the agency continued activities that worked well during the demonstration project, initiated a number of new actions, and is taking steps to address coordination challenges. According to CMS officials, the success of the RAC program depends on collaboration between the RACs and the Medicare claims administration contractors because of the interdependence of their responsibilities. Once the RACs identify errors, Medicare claims administration contractors are responsible for re-processing the claims to repay underpayments or recoup overpayments, conducting the first level review for RAC-related appeals, and informing and training providers about lessons learned through the RAC reviews, according to CMS officials. (See fig. 5 which illustrates this interdependence of RACs and MACs.) CMS is taking multiple steps to resolve RAC and Medicare claims administration contractor coordination issues in the national program based on lessons learned during the demonstration project, such as continuing the RAC and Medicare claims administration contractors vulnerability calls, enhancing the existing data warehouse, automating the claims-adjustment process, and developing a system for electronic documentation sharing when RAC determinations are appealed. CMS is continuing regular RAC and Medicare claims administration contractor vulnerability calls. The vulnerability calls, which began 2 years after the start of the demonstration project, were considered valuable according to agency officials. CMS officials said that they plan to hold weekly calls during the national program, to share RAC-identified vulnerabilities that may result in improper payments with Medicare claims administration contractors. According to CMS, these calls can inform Medicare claims administration contractors about ways to reduce payment errors, for example, by implementing appropriate local system edits or educating providers. CMS noted that conducting these calls during the demonstration project provided information about how best to implement corrective actions that would prevent future improper payments. For example, upon learning about some RAC-identified inpatient hospital errors, CMS consulted coding experts about how to resolve these errors and whether it was necessary to conduct an educational session on the issue. According to a CMS official, the vulnerability calls are expected to serve as the main mechanism of communication between the RACs and the Medicare claims administration contractors about vulnerabilities and are expected to provide a means to share RAC findings with various other components of CMS. CMS is enhancing the data warehouse. For the national program, CMS is redesigning, enhancing, and maintaining the data warehouse created during the demonstration project to house data on RAC activity and prevent RACs from auditing claims under investigation or previously reviewed by other contractors. RACs and one of the Medicare claims administration contractors reported issues with the data warehouse during the demonstration project, including difficulty uploading data in the correct format, slow processing time, and a lack of information on collection activities. According to CMS, it has already made significant changes to the data warehouse. For example, it enhanced the system to accommodate increased user demand, added capability to generate reports for CMS to track RAC activity, and improved processes for data uploads and downloads. CMS also plans to incorporate appeals data into the data warehouse. CMS is automating the claims-adjustment process. According to CMS, the agency is automating the claims-adjustment process to address Medicare claims administration contractors’ workload issues. During the demonstration project, the Medicare claims administration contractors’ workload related to claims adjustment increased significantly, due to the high volume of claims RACs identified that required adjustment and the time-consuming process necessary for the contractors to adjust them. CMS officials stated that the amount of time and effort required of the Medicare claims administration contractors to re-process RAC-related claims was the most significant coordination problem. The agency automated the Part A claims adjustment process and is working to automate the process for adjusting Part B claims by April 2010. CMS officials stated that the changes eliminate the need for costly and time- consuming manual intervention by the Medicare claims administration contractors, ensure that overpayment recovery or underpayment reimbursement occurs promptly, and ultimately minimize the burden on the Medicare claims administration contractors. However, one Medicare claims administration contractor informed us that the Part A claims adjustment process failed to adjust its claims. CMS is developing an electronic documentation sharing system. According to CMS officials, the agency addressed an administrative burden by developing the e-RAC initiative, an electronic system that RACs, CMS, and Medicare claims administration contractors will use to share medical records. CMS officials stated that during the demonstration project, RACs transferred paper copies of medical records to Medicare claims administration contractors for appeals deliberations. According to Medicare claims administration contractors, the volume of appeals made it difficult to manage all of the paper medical records. A CMS official told us the agency expects the first phase of the e-RAC initiative to be operational in March 2010, which would allow the RACs to store imaged files of medical records and make them accessible to CMS and certain contractors that review, but do not process, claims. CMS expects this system to enable the agency to create basic reports and improve oversight of RAC activities. CMS’s goal is to expand the e-RAC initiative to one or more Medicare claims administration contractors by the end of calendar year 2010. CMS established a “black-out period” for claims review. To ensure that the RAC national program does not interfere with the ongoing transition of fiscal intermediaries and carriers to MACs, CMS reported establishing a black out period of three months before and after each transition when the new MACs will focus on other claims processing activities and not work with the RACs in their jurisdictions. Claims processed during this period will be available for RAC review after the black-out period has ended. According to CMS officials, the agency instituted the black-out period, in part, to limit the number of claims adjusted during a time of significant change. CMS is planning to add performance metrics on coordination with RACs into the MAC award fee program. CMS officials indicated that the agency is planning to add performance metrics to provide incentives for coordination between the RACs and MACs into the MAC award fee program. The award fee program is designed to provide incentives for exceptional performance by the MACs. According to CMS officials, these performance metrics will likely include activities such as participating in conference calls; effectively coordinating, implementing, and providing appropriate edit recommendations; and communicating claims determination decisions and inquiries. CMS officials stated that they will add metrics on coordination with the RACs to the award fee program once all of the MACs are in place. CMS Has Taken Steps to Improve Oversight of RAC Accuracy and Service to Providers CMS took a number of steps to improve oversight of the accuracy of RACs’ claims review determinations and the quality of RAC service to providers in the national program. Specifically, CMS added processes to review the accuracy of RAC determinations and established Web site requirements to address provider concerns about service. CMS also established a number of performance metrics to monitor RAC accuracy and service to providers. CMS Established Processes to Review the Accuracy of RAC Determinations and Required Additional RAC Medical Expertise to Enhance Program Accuracy For the national program, CMS created processes to more closely review the accuracy of RAC determinations to address provider concerns raised during the demonstration project. Providers raised concerns that CMS did not sufficiently oversee the RACs during the demonstration project to ensure the vulnerabilities pursued by RACs were valid and that RACs made accurate improper payment determinations. According to provider associations, this led to numerous appeals of inaccurate RAC determinations that were expensive and burdensome for providers. For the national program, CMS will continue a process the agency established during the end of the demonstration project to help ensure that RACs pursue valid vulnerabilities. Prior to pursuing a wide-scale review of any vulnerability, the RAC must submit it to CMS for the agency’s approval. As part of the submission process, the RAC must provide a description of the vulnerability; a reference to the rule, regulation, or policy the RAC intends to evaluate claims against; and a small sample of claims (up to 10) that the RAC already reviewed and the findings for those claims. For example, CMS approved one RAC’s request to identify overpayments associated with providers billing for more than one blood transfusion in a hospital outpatient setting for a Medicare beneficiary in a day—which Medicare policy does not allow. According to CMS officials, the level of review that each proposed vulnerability will receive will depend on its complexity. CMS officials in OFM have authority to allow the RACs to pursue clear-cut vulnerabilities that can lead to improper payments, such as duplicate payments for the same service. For more complex vulnerabilities, including all medical necessity determinations, the agency established a New Issue Review Board, comprised of officials from four CMS components, which will decide whether the RAC can go forward with its proposed review. The board is responsible for ensuring that each RAC’s claims reviews conform to Medicare’s coverage or payment policies and that the language the RAC proposes to use in its determination letters is appropriate and clear. CMS also contracted with a validation contractor (VC) with experience in claims review to independently examine how the RAC plans to select claims for each vulnerability and to determine whether the RAC plans to use the correct review strategy—(automated or complex)—in reviewing claims. In addition, the VC also is expected to reexamine the small sample of claims submitted by RACs with each proposed vulnerability to assess the accuracy of these RAC determinations. In addition to the oversight process for proposed vulnerabilities, CMS also established a process for ongoing oversight of RAC accuracy of the improper payments identified. Each month CMS’s VC is expected to independently examine 100 randomly selected claims that had been reviewed by each RAC. For each claim in the sample, the VC is expected to report whether it agrees or disagrees with the RAC’s determination and evaluate whether the language used by the RAC to communicate the determination to the provider was clear and accurate. CMS officials told us that the agency plans to publish an annual accuracy score for each RAC in the agency’s annual report on the RAC program and will take the scores into consideration when determining whether to renew each RAC’s contract. CMS officials also told us that they may prohibit a RAC with a low score on a particular issue from reviewing additional claims on that issue. This process could help address provider concerns that CMS might not become aware of inaccurate RAC determinations unless providers filed significant numbers of appeals. In addition to these oversight processes, CMS added requirements regarding the medical expertise of RAC staff to help address accuracy concerns. Providers stated that RACs did not have the necessary medical expertise to make their determinations during the demonstration project, because they were not required to have a physician medical director on staff or coding experts conducting the claims reviews. To address this concern, for the national program, CMS required each RAC to have at least one physician on staff as a medical director to provide clinical expertise and judgment to understand Medicare policy, provide guidance in questionable claims review situations, recommend when corrective actions are needed to address the RAC-identified vulnerabilities that result in improper payments, and brief and direct personnel on the correct application of policy during claims review. CMS also required RACs to hire registered nurses or therapists to conduct medical necessity determinations and coding experts to conduct other types of reviews. Providers also reported that CMS’s decision to allow the demonstration RACs to retain contingency fees for determinations overturned at the second through the fifth level of appeal led RACs to make questionable determinations to increase their fees. CMS chose this methodology, in part, to encourage companies to participate in the demonstration project. To address provider concerns about the incentives in the payment method, CMS will require RACs to refund contingency fees received on any determination overturned at any level of the appeals process. CMS Created Web Site Requirements for RACs Designed to Improve Service to Providers In addition to the changes CMS made to improve oversight of RAC accuracy, CMS also created a number of requirements for RAC Web sites to address provider concerns about the RACs’ service. Provider associations reported that during the demonstration project their members could not easily track the status of claims throughout the RAC adjudication process, including the status of medical record request submissions and appeals. CMS also reported in its evaluation report on the RAC demonstration project that providers wanting to track the status of their medical record submissions often had to make frequent phone calls to RAC call centers and read a list of case numbers. CMS required each RAC by January 1, 2010, to develop a tool on its Web site that will allow providers to track the status of a claim. This tool should include information on whether a medical record request is outstanding, whether the RAC received the requested medical records, whether the RAC’s review is underway or complete, and whether the case is closed. As of January 4, 2010, according to a CMS official, providers could track the status of their requested claims on two of the four RAC Web sites. According to a CMS official, the remaining RACs will need to have their tools in place prior to issuing requests for medical records. Although providers expressed concern about the difficulty tracking the status of their appeals during the demonstration project, CMS has not required the RAC Web sites to include information on the status of appeals resulting from RAC determinations. According to CMS officials, the agency does not have a standard system to track first-level appeals, and it would be difficult for RACs to collect the information from a number of separate Medicare claims administration contractors. CMS officials overseeing the RAC program told us they are working with their counterparts in the Medicare appeals division within CMS to move up the date by which the Medicare claims administration contractors will begin using the CMS system that already tracks appeals at the second and third level. These same officials told us they anticipate RACs will eventually incorporate appeals information into their Web sites, though the inclusion of appeals information is not a requirement in the RAC contract. Providers also expressed concern that they did not know what vulnerabilities RACs were pursuing during the demonstration project. In addition to the new issue review process, CMS has required the RACs to post a description of each vulnerability that they audit on their Web sites. The postings include a description of the vulnerability, the states where the RAC identified the problem, and references to additional information about the vulnerability. According to CMS officials, providers will need to check the Web site of the RAC in their region to stay informed of emerging vulnerabilities under RAC review for improper payments. To address provider concerns about medical record requests getting lost during the demonstration project because a RAC did not send the request to the correct department or individual at a hospital or practice, CMS is requiring each RAC to develop a tool for its Web sites that will allow providers to customize their address and point-of-contact information. CMS also encouraged the RACs to solicit the assistance of provider associations to help collect the information. CMS Developed Performance Metrics to Monitor RAC Accuracy and Provider Service CMS developed performance metrics to oversee RAC accuracy, service to providers, and other aspects of performance. The performance metrics include measurements of the RACs’ compliance with medical record request limits and the accuracy of RAC determinations, as evaluated by the VC, as well as measures of staff performance at each RAC’s customer service phone number that is expected to respond to inquiries from providers. (See table 1.) CMS’s RAC project officers will be responsible for monitoring each RAC’s performance and following up with the RAC if its performance does not meet the required level in the national program. For instance, to monitor whether call center staff answer questions fully and accurately, project officers or their designees will randomly monitor calls to the RAC call center and investigate provider complaints. If a project officer determines that call center staff are not answering questions fully and completely all the time, the project officer will require the RAC to respond in writing to the finding and may require a corrective action plan. CMS’s statement of work also includes a provision that CMS may stop recovery work in a particular region if evidence leads CMS to believe the RAC’s plan to provide service to providers is inappropriate or ineffective. In such a case, CMS would not allow the RAC to resume recovery work until the RAC satisfied CMS it made all required improvements to its provider service in the area. Conclusions The ultimate success of the government-wide effort to reduce improper payments hinges on each federal agency’s diligence and commitment to identify, estimate, determine the causes of, take corrective actions on, and measure progress in reducing improper payments. To this end, CMS must establish effective accountability measures, and incentives, to ensure the RAC program meets the agency’s stated objectives. Although the RAC demonstration project led to the successful recoupment and refunding of past improper payments, CMS did not focus sufficient attention on addressing the root causes of the vulnerabilities that caused them. Neither the IPPP developed during the demonstration project nor the current plan for the national program provide for sufficient monitoring and control activities to ensure that corrective actions are taken to help meet the overall goal of reducing improper payments in the Medicare program. Because the RAC national program team does not have the organizational authority within the agency to implement the corrective actions needed to address the vulnerabilities that lead to improper payments, CMS must develop criteria by which it prioritizes the activities of its various components and contractors to develop adequate measures to reduce future improper payments. The identification and prevention of future Medicare FFS improper payments due to vulnerabilities identified by the national RAC program require direction from a sufficiently high level within CMS to initiate action from the various parts of the agency and its contractors. In addition, assessing the effectiveness of the corrective actions taken is an important step for reducing future improper payments. Recommendations for Executive Action To help reduce future improper payments, we recommend that the Administrator of CMS develop and implement a process that includes policies and procedures to ensure that the agency promptly: evaluates findings of RAC audits, decides on the appropriate response and a time frame for taking action based on established criteria, and acts to correct the vulnerabilities identified. As part of this process, we recommend that the Administrator of CMS designate key personnel with appropriate authority to be responsible for ensuring that corrective actions are implemented and that the actions taken were effective. Agency and Other External Comments We provided a draft of this report to the HHS for comment. We also provided statements of facts from our draft report to the two Medicare claims administration contractors and seven provider associations we interviewed and requested their comments. We received written comments from HHS on behalf of CMS. These comments are reprinted in Appendix II. We also received oral or written comments from two Medicare claims administration contractors and five of the seven provider associations on statements of facts related to information they provided, including some technical comments that we incorporated as appropriate. CMS Comments CMS commented that the national RAC program is an important step in meeting its commitment to lower the Medicare payment error rate. The agency indicated that our review imparted vital recommendations that will greatly enhance CMS’s oversight of the RAC national program and CMS concurred with each of our recommendations. With regard to the recommendation that CMS promptly evaluate the findings of RAC audits, CMS concurred and discussed specific elements included in the national program that are designed to report vulnerabilities from RAC audits and potential corrective actions. CMS concurred with our recommendation that the agency implement a process to decide on the appropriate response to address each RAC-identified vulnerability, but indicated that more research might be needed to determine the appropriate response or corrective action for some vulnerabilities. CMS also concurred that the agency should act promptly to correct the vulnerabilities, but indicated that it did not consider a vulnerability to be validated until the majority of claims for that issue completed the Medicare appeals process. Since the appeals process can take more than 2 years, the approach CMS suggested in its comments did not align with the intent of our recommendation. After conferring with CMS officials to clarify the agency’s intent on acting promptly on vulnerabilities identified during the RAC national program, CMS acknowledged that it intended to review vulnerabilities on a case-by- case basis and judge how quickly to act on each. Agency officials told us they were considering assigning vulnerabilities to risk categories from high to low that would help to determine whether the agency should take prompt action or whether it should wait for claims to complete the appeals process. These officials told us that waiting for the results of appeals would keep the agency from expending the resources on corrective actions that would need to be reversed if the appeals process overruled RAC determinations. We agree that taking a risk-based approach meets the intent of the recommendation. To clarify this intent, we modified our recommendation to make the prompt prioritizing and timing of corrective actions, based on established criteria, more explicit. Finally, CMS concurred with our recommendation that the agency designate key personnel to oversee that corrective actions are implemented and effective and stated that the Administrator of CMS is the official responsible for assuring that vulnerabilities that cut across all agency components are addressed. Other External Comments We clarified information in the report based on comments from two Medicare claims administration contractors. In addition, the five associations that provided comments to us did not offer substantive changes to the statement of facts that they reviewed. Three associations affirmed that the draft report addressed issues they had raised about the RAC demonstration project and national program. These three associations also discussed in greater detail concerns that they continue to have with the RAC program, such as the many appeals still in process from the RAC demonstration project. The other two provider associations raised no substantive issues with the report. We are sending copies of this report to the Administrator of CMS and other interested parties. In addition, the report will be available at no charge on GAO’s Web site at http://www.gao.gov. Please contact us on (202) 512-7114 or (202) 512-9095 if you or your staff have any questions about this report. Contact points for our Office of Congressional Relations and Office of Public Affairs can be found on the last page of this report. Other major contributors to this report are listed in Appendix III. Appendix I: Selected Changes Made to the Medicare National Recovery Audit Contractors (RAC) Program As a result of the RAC demonstration project, the Centers for Medicare & Medicaid Services (CMS) included the following features in the RAC national program: RACs are to have a physician medical director. RACs are to be staffed with registered nurses or therapists to make coverage and medical necessity determinations and certified coders to make coding determinations. RACs are to make credentials of reviewers available to providers upon request. Providers will be able to discuss claim denials with the RAC medical director upon request. The minimum claim amount that the RACs will review was raised to $10 minimum per claim (instead of $10 minimum for aggregated claims). CMS will use a validation contractor to independently examine the criteria each RAC plans to use to make its determinations and the accuracy of RAC determinations. RACs must return the related contingency fee if a claim is overturned on appeal. RACs must use standardized letters to notify providers of overpayments The look-back period (from claim payment date to date of medical record request) is reduced from 4 years to 3 years. The RACs are allowed to review claims paid in the current fiscal year. CMS is putting limits on the number of medical record requests in a 45 day period. The time frame for paying hospital medical record photocopying vouchers is to be set at 45 days from receipt of medical record. CMS is not including Medicare Secondary Payer claims audits in the National Program. RACs are to have quality assurance/ internal control audits. RACs are to list the reason for review on “request for records” letters and overpayment letters. The status of specific claims are to be posted on RAC Web page. RAC contingency fees are to be made publicly available. Appendix II: Comments from the Department of Health & Human Services Appendix III: GAO Contacts and Staff Acknowledgments GAO Contacts Acknowledgments In addition to the contacts named above, Sheila K. Avruch, Assistant Director; Carla Lewis, Assistant Director; Lori Achman; Jennie Apter; Anne Hopewell; Nina M. Rostro; and Jennifer Saunders made key contributions to this report.

Summary: The Centers for Medicare & Medicaid Services (CMS) conducted a mandated 3-year project from March 2005 through March 2008 to demonstrate the use of recovery audit contractors (RAC) in identifying Medicare improper payments and recouping overpayments. CMS implemented a mandated national RAC program, which began in March 2009. GAO was asked to examine specific issues that arose during the demonstration project and CMS's efforts to address them in the national RAC program. This report examines the extent to which CMS (1) developed a process and took corrective actions to address vulnerabilities identified by the RACs that led to improper payments, (2) resolved coordination issues between the RACs and the Medicare claims administration contractors, and (3) established methods to oversee RAC claim review accuracy and provider service during the national program. GAO reviewed CMS documents and interviewed officials from CMS and contractors and provider groups affected by the demonstration project. CMS did not establish an adequate process in the 3-year demonstration project or in planning for the national program to address RAC-identified vulnerabilities that led to improper payments, such as paying duplicate claims for the same service. CMS stated that one purpose of the demonstration project was to obtain information to help prevent improper payments. However, CMS has not yet implemented corrective actions for 60 percent of the most significant RAC-identified vulnerabilities that led to improper payments, a situation that left 35 of 58 unaddressed. These were vulnerabilities for which RACs identified over $1 million in improper payments for medical services or $500,000 for durable medical equipment. CMS developed a spreadsheet, which listed the most significant improper payment vulnerabilities that were identified by the RACs during the demonstration project. However, the agency did not develop a plan to take corrective action or implement sufficient monitoring, oversight, and control activities to ensure these significant vulnerabilities were addressed. Thus, CMS did not address significant vulnerabilities representing $231 million in overpayments identified by the RACs during the demonstration project. For the RAC national program, CMS developed a process to compile identified vulnerabilities and recommend actions to prevent improper payments. However, this corrective action process lacks certain essential procedures and staff with the authority to ensure that these vulnerabilities are resolved promptly and adequately to prevent further improper payments. Based on lessons learned during the demonstration project, CMS took multiple steps in the national program to resolve coordination issues between the RACs and Medicare claims administration contractors. During the demonstration project, CMS learned that having regular communication with the claims administration contractors on improper payment vulnerabilities that the RACs were identifying was important. CMS also learned that the data warehouse used to store claims information for the RACs needed more capacity and utility, that manual claims adjustment by claims administration contractors to recoup improper payments was burdensome, and that sharing paper copies of medical records between RACs and claims administration contractors when claims denials were appealed was difficult to manage. As a result, CMS took steps to resolve these coordination issues in the national program, such as enhancing the existing data warehouse and automating the claims-adjustment process. CMS took steps to improve oversight of the accuracy of RACs' claims reviews and the quality of their service to providers for the national program. CMS added processes to review the accuracy of RAC determinations, including independent reviews by another CMS contractor. CMS also established requirements to address provider concerns about service, such as having the RACs establish Web sites that will allow providers to track the status of a claim being reviewed. In addition, CMS established performance metrics that the agency will use to monitor RAC accuracy and service to providers.

gov_report

en

Summarize: The family of American photojournalist Luke Somers held captive in Yemen broke their silence Thursday, releasing a video to plead for mercy after US officials confirmed they had tried and failed to rescue him. The brief YouTube clip featuring Somers’ brother, Jordan, and his mother, Paula Somers, came out just hours after al Qaeda's Yemeni affiliate released their own video, threatening to kill the British-born journalist in three days' time unless the US government meets several demands. Somers, 33, was captured on September 17, 2013, in Yemen's capital of Sana'a. Scroll down for videos. Al Qaeda in the Arabian Peninsula said it will murder British-born hostage Luke Somers, pictured, within three days unless the United States agrees to a range of demands. Mr Somers was kidnapped in Yemen. Begging for mercy: Luke's brother, Jordan (left), and his mother, Paula Somers (right), released a video Thursday asking his al Qaeda captors to release him. 'Up to this point, we have no explanation as to why Luke was targeted as a victim, and we currently don't know why he's being held,' Jordan Somers says into the camera, directly addressing his brother's captors. 'Luke has spent the last two years making Yemen his home. He's a good person and he's only been trying to do good things for the Yemeni population. 'He goes out of his way to care for, and respects, the common person and he has made many lasting friends in Yemen.' Jordan Somers then goes on to claim that their family were unaware of the failed special operations raid on a cave in Yemen intended to liberate Luke. 'Luke is only a photojournalist and he is not responsible for any actions the US government has taken,' he says. 'Please understand that we had no prior knowledge of the rescue attempt for Luke, and we mean no harm to anyone.' The hostage's mother, Paula Somers, then speaks, thanking the al Qaeda group for taking a good care of Luke and keeping him healthy. 'Please, show mercy and give us an opportunity to see our Luke again; he is all that we have,' she says into the camera. 'Luke, if you are able to hear or see us, please know that we're doing everything possible to help you. Our hearts are with you, we miss you and we love you, and all we want to do is see you again and have you safely in our arms, all together again,' the mother concludes. Pentagon spokesman Rear Admiral John Kirby said in a statement Thursday that eight days ago, special forces raided a cave complex in Yemen to free a number of hostages. Some of the captives were rescued, but others, among them Somers, were not present at the targeted location. Nasser bin Ali al Ansi, senior official in Al Qaeda in the Arabian Peninsula, pictured, spoke for two minutes and thirty seconds during the video where he threatened to kill Mr Somers within three days. White House spokesman Bernadette Meehan said the mission was coordinated with the Yemeni government and carried out by a group including both American special operations forces and Yemeni military. 'The overriding concern for Mr. Somers' safety and the safety of the U.S. forces who undertake these missions made it imperative that we not disclose information related to Mr. Somers' captivity and the attempted rescue,' Meehan said, adding that the White House's 'thoughts remain with the Somers family. Meehan said the mission was being disclosed now because of the video released Thursday. The video was released this morning by the Al Malahem Media Center, Al Qaeda in the Arabian Peninsula's propaganda arm, and discovered by the SITE Intelligence Group which monitors terrorist activity online. Mr Somers moved from London to Sana'a, Yemen in 2011 to become a teacher, but soon started taking pictures of public demonstrations and established himself as a photojournalist working for the Yemen Times. In this Monday, February 11, 2013 photo, Luke Somers poses for a picture during a parade marking the second anniversary of the revolution in Sanaa, Yemen. The broadcast features Nasser bin Ali al Ansi, who is a senior official in Al Qaeda in the Arabian Peninsula (AQAP). Ansi warns President Obama that unless he accepts the terror group's demands, 'the American hostage held by us will meet his inevitable fate'. During the video, Ansi does not cover his face and is seen reading from a script in Arabic which accuses America of committing crimes against Muslims. Ansi remains vague about what specific demands need to be met by the U.S. government to ensure Somers' release, but says the Americans are 'aware' of the list. Unlike previous videos produced by ISIS and the Al Hayat Media Center, Mr Somers is not wearing an orange jump suit nor is he threatened with any form of weapon on screen. During the video, Mr Somers addresses the camera, saying: 'My name is Luke Somers. I'm 33 years old. I was born in England, but I carry American citizenship and have lived in America for most of my life. 'It's now been well over a year since I've been kidnapped in Sana'a. Basically, I'm looking for any help that can get me out of this situation. I'm certain that my life is in danger. 'So as I sit here now, I ask if anything can be done, please let it be done. Thank you very much.' 'It's now been well over a year since I've been kidnapped in Sana'a. Basically, I'm looking for any help that can get me out of this situation. I'm certain that my life is in danger,' Mr Somers says in the video. American commandos tried to rescue Mr Somers in a raid on an AQAP camp late last month, but he had been moved by the time they arrived. The video was released eight days after American commandos carried out a raid on an AQAP camp, in an attempt to rescue Mr Somers. The raid took place on a cave complex in the remote Hadhramaut province where locals give terrorist cells safe haven. But by the time members of SEAL Team Six, the elite unit that killed Osama Bin Laden in 2011, raided the AQAP camp on November 26, he had already been moved. However, the special forces recovered a group of eight hostages including six Yemenis, a Saudi Arabian and an Ethiopian. One of the hostages rescued in the raid allegedly reported that Mr Somers may have been moved out with another group of prisoners two days before the raid took place. Mr Somers was believed to be a part of a group which included citizens from Britain, South Africa and Turkey. The New York Times reports that A Yemeni electricity company worker named Rasheed al-Habishi was also expected to be in Mr Somers' camp. On Thursday, his son told the Times that his father's dead body had been found in a town near where the raid took place. The Pentagon initially asked members of the media not to report on the raid, for fear it would jeopardize Mr Somers' life, but the story leaked anyway. It is understood that Mr Somers moved to Sana'a from London in 2010 to work as a teacher. However, he soon started taken photographs of anti-government protests in the city and established himself as a photojournalist working for the Yemen Times. Mr Somers was kidnapped in the Yemeni capital Sana'a in September 2013 by an Al Qaeda affiliate. Mr Somers also worked as a translator and as a freelance photographer, with his pictures appearing on websites like the BBC. The National Yemen newspaper published an article about Mr Somers this week, saying he was 'known as the most active photojournalist at Change Square' - a protest site at the center of Yemen's 2011 rebellion against former President Ali Abdullah Saleh. However, not much else has been said about Mr Somers, especially about his life before moving to Yemen. The U.S. government tries not to publicize the kidnapping of citizens, in an attempt to shield them from hostage situations like this. The three-minute video also features Ansi speaking about American activity in Afghanistan, Somalia and Iraq as well as recent air strikes in Syria. It follows similar videos by ISIS, which has already killed two British hostages and three American hostages in videos released on social media. Rival terror group ISIS has threatened to murder British journalist John Cantlie, pictured. ISIS has posted a series of videos online showing the separate murders of U.S. journalists James Foley and Steven Sotloff, U.S. aid worker Peter Kassig and two British aid workers, David Haines and Alan Henning. Footage claiming to show Mr Henning's murder appeared on the internet just days after the UK joined US-led air strikes against the terrorists in Iraq. And official figures suggest around 500 Britons have travelled to Syria and Iraq to fight for ISIS, while others have joined up with Kurdish groups to fight against the militants. The government last month announced a raft of new anti-extremist measures, including ensuring insurance companies can no longer foot the bill for terrorist ransoms, suspected foreign fighters will be blocked from returning to the UK and powers will be reintroduced to relocate terror suspects across the country

Summary: Luke Somers, 33, was kidnapped in the Yemeni capital in September 2013. Al Qaeda in the Arabian Peninsula has issued a three-day deadline. Terrorists said they will kill Mr Somers if the US does not accept demands. Mr Somers was born in the UK but is now an American citizen. He said: 'I'm looking for any help that can get me out of this situation' SEAL Team Six raided a remote Yemeni terror camp on November 25. The commandos rescued some Yemeni and Saudi hostages. But Mr Somers had been moved out of the camp two days earlier.

cnn_dailymail

en

Write a title and summarize: CD8 T cell responses have three phases: expansion, contraction, and memory. Dynamic alterations in proliferation and apoptotic rates control CD8 T cell numbers at each phase, which in turn dictate the magnitude of CD8 T cell memory. Identification of signaling pathways that control CD8 T cell memory is incomplete. The PI3K/Akt signaling pathway controls cell growth in many cell types by modulating the activity of FOXO transcription factors. But the role of FOXOs in regulating CD8 T cell memory remains unknown. We show that phosphorylation of Akt, FOXO and mTOR in CD8 T cells occurs in a dynamic fashion in vivo during an acute viral infection. To elucidate the potentially dynamic role for FOXO3 in regulating homeostasis of activated CD8 T cells in lymphoid and non-lymphoid organs, we infected global and T cell-specific FOXO3-deficient mice with Lymphocytic Choriomeningitis Virus (LCMV). We found that FOXO3 deficiency induced a marked increase in the expansion of effector CD8 T cells, preferentially in the spleen, by T cell-intrinsic mechanisms. Mechanistically, the enhanced accumulation of proliferating CD8 T cells in FOXO3-deficient mice was not attributed to an augmented rate of cell division, but instead was linked to a reduction in cellular apoptosis. These data suggested that FOXO3 might inhibit accumulation of growth factor-deprived proliferating CD8 T cells by reducing their viability. By virtue of greater accumulation of memory precursor effector cells during expansion, the numbers of memory CD8 T cells were strikingly increased in the spleens of both global and T cell-specific FOXO3-deficient mice. The augmented CD8 T cell memory was durable, and FOXO3 deficiency did not perturb any of the qualitative attributes of memory T cells. In summary, we have identified FOXO3 as a critical regulator of CD8 T cell memory, and therapeutic modulation of FOXO3 might enhance vaccine-induced protective immunity against intracellular pathogens. The ability of the immune system to respond rapidly and vigorously to antigen re-exposure is termed immunological memory, which is one of the tenets of adaptive immunity [1], [2]. Induction of memory B and T cells is the basis of immunological memory induced by infections or vaccinations [2]–[4]. As compared to naïve T cells, memory T cells are hyper-reactive to antigenic stimulation and swiftly proliferate and/or differentiate into effector cells to confer protective immunity expeditiously [5]–[8]. The ability of memory T cells to confer protective immunity depends upon the number and quality of memory T cells [5], [9]–[13]. Understanding the mechanisms that regulate the quantity and quality of T cell memory is fundamentally important for the development of effective vaccines. During a CD8 T cell response, engagement of the TCR, along with appropriate co-stimulatory and inflammatory signals, activate naïve T cells to proliferate and differentiate into effector cells [1], [4], [8], [13], [14]. In the case of LCMV infection, the peak of T cell expansion is reached at 8–10 days after infection, and the majority of the newly generated effector cells present at the peak of the response are short-lived and fated for deletion [15]–[17]. But, a small subset of the effectors, termed memory precursor effector cells (MPECs), possesses the potential to survive and differentiate into long-lived memory cells [16], [17]. The number of memory CD8 T cells generated depends largely upon the magnitude of the expansion of MPECs during the T cell response. Substantial progress has been made in deciphering the extracellular signals and transcription factors that regulate the differentiation of MPECs [1], but the signaling pathways that govern the number of MPECs, their differentiation into memory CD8 T cells, and the maintenance of CD8 T cell memory are not fully understood. The FOXO family of transcription factors plays a crucial role in governing cellular proliferation, apoptosis, energy metabolism, and stress resistance in response to dynamic alterations in stress and abundance of nutrients and growth factors in many cell types [18]–[26]. In mammals, the FOXO family consists of at least four members: FOXO1, FOXO3, FOXO4, and FOXO6 [22], [27], [28]. The activity of FOXOs is regulated by post-translational modifications, most notably, phosphorylation [25], [26], [29]–[31]. In resting, quiescent cells, FOXOs exist in a hypo-phosphorylated state and localize to the nucleus to control the transcription of their target genes such as p27Kip1, p21Cip1, p300, BIM, Fas ligand that are involved in regulating cellular proliferation or apoptosis [25], [32]–[43]. However, in response to stimulation with growth factors or cytokines that stimulate the PI3K/AKT signaling pathway, activated Akt phosphorylates FOXO leading to its exclusion from the nucleus and degradation by proteolysis in the cytoplasm [18], [25], [29], [44], [45]. As a result, the transcription of FOXO target genes is diminished, which in turn facilitates cell cycle entry and/or survival. T cells express FOXO1 and FOXO3, and there has been a recent surge in interest to elucidate the importance of FOXOs in regulating T cell biology [46]–[53]. Phosphorylation-mediated inactivation of FOXOs and downregulation of p27Kip1 appear to be obligatory steps for T cells to enter the cell cycle in response to TCR engagement [24], [41], [53]. FOXO1 has been shown to control several aspects of T cells including the expression of adhesion molecules like L-selectin and CCR7, cytokine receptors like the IL-7 receptor, development of regulatory T cells, and protection against autoimmunity [51], [54]. Unlike FOXO1, the role of FOXO3 in T cell homeostasis is less well understood. It was first reported that global deletion of FOXO3 results in lymphadenopathy and spontaneous activation of T cells, but other independent studies have failed to confirm these results, possibly due to differences in the genetic background of mutant mice [19], [40], [49]. Nonetheless, elegant studies from the Hedrick group have showed that FOXO3 inhibits the primary expansion of CD8 T cells in the spleen by regulating IL-6 production by dendritic cells [49]. However, the T cell intrinsic role of FOXO3 in regulating various phases of the polyclonal multi-epitope-specific CD8 T cell response to an acute viral infection in lymphoid and non-lymphoid organs remains to be determined. In this study, using global and conditional T cell-specific FOXO3 knockout mice, we have systematically examined the role of FOXO3 in regulating the: (1) expansion and function of polyclonal antigen-specific CD8 T cells in lymphoid and non-lymphoid organs; (2) antigen-driven in vivo proliferation of virus-specific CD8 T cells; (3) differentiation of CD8 T cells into short-lived effector cells (SLECs) and MPECs; (4) contraction of antigen-specific CD8 T cells in lymphoid and non-lymphoid organs; (5) the numbers and function of memory CD8 T cells; (6) proliferative renewal of memory CD8 T cells; (7) secondary CD8 T cell responses and protective immunity. These studies show that FOXO3 regulates the clonal expansion of polyclonal CD8 T cells in a tissue-specific fashion by T cell intrinsic mechanisms. The enhanced expansion of CD8 T cells was clearly not due to an increased proliferation rate, but was instead associated with reduced cellular apoptosis. Furthermore, we show that FOXO3 deficiency markedly enhances the size of the memory CD8 T cell compartment without affecting the phenotype or quality of memory CD8 T cells. These findings have implications in the design of effective vaccines that engender potent and effective protective cellular immunity against intracellular pathogens and tumors. FOXO3 has emerged as a key regulator in a number of physiological outcomes, including metabolism, ageing, and vascular reactivity [21]–[23], [27], [46], [55]. More recently, in the immune system, there is increasing evidence that FOXO3 is a critical regulator of T cell homeostasis [46], [47], [52], [53], [55]. While a number of signaling pathways and post translational modifications may be involved in controlling the activity of FOXO3, phosphorylation mediated through a PI3K-Akt centric signaling module primarily regulates FOXO3 function in T cells [29], [45], [56], [57]. In order to fully understand the role of the Akt/FOXO3 axis in the control of CD8 T cell homeostasis, we developed a phospho-specific, flow cytometric method to quantify in vivo phosphorylation levels of key proteins implicated in FOXO3 activity: the upstream kinase Akt, FOXO3 itself, as well as a potential downstream substrate of Akt, the kinase, mammalian target of Rapamycin (mTOR) (Figure 1A). Figure 1A illustrates the kinetics of phosphorylation of Akt (Thr308), FOXO1/3 (T24/T32), and mTOR (S2448) in antigen-specific CD8 T cells during an acute LCMV infection. The phosphorylation of Akt (Thr308) and mTOR (S2448) was highest at day 5 post-infection (PI), and subsided to steady-state levels by days 10 and 8 PI respectively. Interestingly, the phosphorylation dynamics for FOXO1/O3 were different from that of Akt and mTOR; the phosphorylation levels for FOXO1/O3 dropped between days 5 and 8 PI, but gradually increased back to steady-state levels by days 10–15 PI. Note that the phosphorylation kinetics of NP396-specific CD8 T cells after day 8 PI was slightly delayed, as compared to those in GP33-specific CD8 T cells. In summary, these findings demonstrate that the in vivo phosphorylation levels of Akt, FOXO1/O3, and mTOR are highly dynamic, and of note, the phosphorylation kinetics of Akt correlated with mTOR phosphorylation but not with phosphorylation of FOXO1/O3 during a T cell response to LCMV. However, the specific role of FOXO3 in regulating different phases of the CD8 T cell response has not been carefully examined. Infection of mice with LCMV elicits a potent, multiple epitope-specific, CD8 T cell response wherein virus-specific CD8 T cells are distributed to both lymphoid and non-lymphoid organs [58], [59]. To examine the role of FOXO3 in regulating CD8 T cell responses to an acute viral infection, groups of wild type (+/+) and FOXO3-deficient (FOXO3−/−) mice were infected with LCMV. At day 8 PI, we quantified CD8 T cells that are specific to three immuno-dominant LCMV epitopes in lymphoid (spleen and lymph nodes) and non-lymphoid (liver) tissues. Figure 1B shows that the numbers of LCMV-specific CD8 T cells in spleens of FOXO3−/− mice were significantly (P<0. 05) higher than in +/+ mice. Surprisingly, the numbers of virus-specific CD8 T cells in the lymph nodes and liver of FOXO3−/− mice were comparable to those in +/+ mice (Figure 1B). These data suggested that FOXO3 downregulates the accumulation of CD8 T cells in a tissue-specific fashion during an acute LCMV infection. Next, we assessed whether FOXO3 deficiency affected the cell surface phenotype of LCMV-specific effector CD8 T cells at day 8 PI. The expression levels of CD44, CD62L, CD27, and CD122 on FOXO3−/− LCMV-specific CD8 T cells were comparable to those on +/+ CD8 T cells (Figure 2A). The population of LCMV-specific CD8 T cells in the spleen is comprised of at least two subsets of effector cells based on the cell surface expression of IL-7 receptor α (CD127) and KLRG-1: the SLECs (KLRG-1HI/ CD127LO) a majority of which are destined for apoptosis, and the MPECs (CD127HI/KLRG-1LO), which are poised to differentiate into memory CD8 T cells [16], [17], [60]. Figure 2B shows that FOXO3 deficiency significantly enhanced the absolute numbers of both SLECs and MPECs in the spleen at day 8 PI. Of note is the marked increase in the total number of MPECs in FOXO3−/− mice. In summary, these data suggested that FOXO3 deficiency increased the clonal expansion of CD8 T cells without disrupting the differentiation of SLECs and MPECs. To detect possible differences in the functionality of antigen-specific CD8 T cells from +/+ and FOXO3−/− mice, we assessed their ability to produce the cytokines IFNγ, TNFα, and IL-2 in response to antigenic stimulation directly ex vivo (Figure 2C). The MFIs of IFNγ staining for FOXO3−/− and +/+ effector CD8 T cells were similar (Figure 2C). Additionally, the percentages of epitope-specific, CD8 T cells that produced two cytokines (IFNγ and TNFα) or three cytokines (IFNγ, TNFα, and IL-2) in FOXO3−/− mice were similar to those in +/+ mice (Figure 2C). The MFIs for TNFα and IL-2 were also comparable between +/+ and FOXO3−/− CD8 T cells (data not shown). As a surrogate marker of the lytic function of effector CD8 T cells, we compared granzyme B expression between +/+ and FOXO3−/− LCMV-specific effector CD8 T cells directly ex vivo. The levels of granzyme B in FOXO3−/− CD8 T cells were similar to those in +/+ CD8 T cells (Figure 2D). Taken together, data in Figures 1 and 2 suggested that FOXO3 deficiency increased the expansion of virus-specific CD8 T cells without affecting their phenotype or function. Consistent with normal CD8 T cell effector function, LCMV control in FOXO3−/− mice was similar to that in +/+ mice (not shown). To rigorously address whether the greater number of LCMV-specific CD8 T cells in the spleen of FOXO3−/− mice was due to increased proliferation, we utilized two different approaches, Ki67 staining and BrdU incorporation. First, we quantified proliferation of LCMV-specific CD8 T cells by staining for the nuclear antigen Ki67 directly ex vivo (Figure 3A) during the peak clonal expansion phase (days 6 and 8 PI) of the CD8 T cell response. For the second approach, we measured BrdU incorporation by LCMV-specific CD8 T cells in vivo from days 6–8 PI (Figure 3B). As shown in Figures 3A and 3B, the percentages of of Ki67+ve LCMV-specific CD8 T cells and BrdU+ve LCMV epitope-specific CD8 T cells in spleens, liver, and lymph nodes of FOXO3−/− mice were similar to those in +/+ mice. Measurement of Ki67 expression at day 5 PI also showed that FOXO3 deficiency did not alter the proliferation of LCMV-specific CD8 T cells (Figure S1) Taken together, these data indicated that enhanced proliferation was not sufficient to explain the increase in antigen-specific CD8 T cells in FOXO3−/− mice during LCMV infection. CD8 T cell homeostasis is not simply determined by alterations in proliferation, it is also regulated by alterations in cell death. During the clonal expansion phase of the CD8 T cell response, there is concomitant proliferation and apoptosis [61], and therefore the magnitude of clonal expansion is dependent upon the relative rates of proliferation and apoptosis. To assess cellular apoptosis, we determined the percentages of Annexin VHI LCMV-specific CD8 T cells in spleens of +/+ and FOXO3−/− mice at day 6 PI, directly ex vivo. Data in Figure 4A shows that the percentages of Annexin VHI LCMV-specific CD8 T cells in spleens of FOXO3−/− mice were significantly lower than in spleens of +/+ mice. These data suggested that FOXO3 controls the accumulation of effector CD8 T cells by promoting cellular apoptosis during an acute LCMV infection. During antigen-driven proliferation, the competing pro-apoptotic effects of TGF-β and the anti-apoptotic effects of IL-15 regulate the apoptotic rate of CD8 T cells [61]. Because apoptosis of LCMV-specific CD8 T cells was dampened by FOXO3 deficiency (Figure 4A), and IL-15 has been reported to induce phosphorylation of FOXO3 [62], we hypothesized that FOXO3 might downregulate clonal expansion by reducing the viability of CD8 T cells that are deprived of IL-15. The balance in the levels of the anti-apoptotic molecule Bcl-2 and the pro-apoptotic molecule BIM controls the susceptibility of a T cell to apoptotic stimuli [36], [38], [39], [63]–[65]. Since BIM is a target gene for FOXO3, we tested whether deficiency of FOXO3 might lead to lower expression of BIM in proliferating CD8 T cells (day 6 PI), by comparing the levels of BIM in +/+ and FOXO3−/− LCMV-specific CD8 T cells after culture with or without IL-15. As shown in Figure 4B and Figure S2, BIM expression in +/+ CD8 T cells was higher than in FOXO3−/− CD8 T cells, when cultured in media without IL-15. However, IL-15 reduced BIM expression in +/+ CD8 T cells to levels seen in FOXO3−/− CD8 T cells; IL-15 did not affect BIM expression in FOXO3−/− CD8 T cells. These data suggested that FOXO3 might control BIM expression in IL-15-deprived CD8 T cells. When analyzing for BIM in direct relation to Bcl-2, we observed that after 6 days of infection, LCMV-specific CD8 T cells from +/+ mice exhibit an increased BIM to Bcl-2 ratio, as compared to FOXO3−/− CD8 T cells (Figure 4B). Thus, we propose that FOXO3 might downregulate the accumulation of proliferating CD8 T cells by inducing BIM expression. Next, we examined whether FOXO3 deficiency regulated the contraction of CD8 T cells in lymphoid and non-lymphoid tissues during an acute LCMV infection. Virus-specific CD8 T cells were quantified in spleen, liver, and lymph nodes at days 8,11,15, and 30 PI (Figure 5A). Overall, the slopes of the contraction curves for LCMV-specific CD8 T cells in FOXO3−/− mice were comparable to those of +/+ mice. Thus, the contraction of LCMV-specific CD8 T cells was minimally affected by FOXO3 deficiency. Next, we assessed whether FOXO3 regulated proliferation of LCMV-specific CD8 T cells during the contraction phase. In vivo BrdU incorporation studies showed that the percentages of LCMV-specific CD8 T cells that incorporated BrdU between days 8–11 or 12–15 PI in FOXO3−/− mice were similar to those in +/+ mice (Figure 5B). The number of memory CD8 T cells is a function of the magnitude of expansion and contraction during the T cell response [5]. Here, we determined whether increased clonal expansion of MPECs in FOXO3−/− mice (Figure 2B) translated to inflation of LCMV-specific memory CD8 T cells in lymphoid and non-lymphoid organs. We observed that FOXO3−/− mice exhibit a substantial increase in the numbers of NP396- (P<0. 001), GP33- (P<0. 01), and GP276- (P<0. 04) specific CD8 T cells in spleen at 180 days PI (Figure 6A). Interestingly, there was no detectable increase in the numbers of LCMV-specific memory CD8 T cells in either the liver or lymph nodes at day 180 PI (Figure 6A). It should be noted that the magnitude of increase in the number of memory CD8 T cells in FOXO3−/− mice reflected the increased accumulation of MPECs during the primary response (Figure 2B). High numbers of memory CD8 T cells were maintained in FOXO3−/− mice stably until at least day 300 PI (data not shown). These data strongly imply that FOXO3 plays an important role in downregulating the magnitude of CD8 T cell memory in the spleen following an acute viral infection. Phenotypic analysis of LCMV-specific memory CD8 T cells in FOXO3−/− and +/+ mice suggested that FOXO3 deficiency did not affect the expression of molecules that control T cell trafficking (CD44) or cytokine receptors (CD122 and CD127) (Figure 6B). In addition, assessment of CD62L levels on antigen specific CD8 T cells indicated that both central (CD62Lhigh) and effector (CD62Llow) memory frequencies were unaffected by FOXO3 deficiency (Figure 6C). Furthermore, functional analysis of antigen-triggered cytokine production did not reveal alterations in cytokine producing ability of LCMV-specific memory CD8 T cells from FOXO3−/− mice when compared to their +/+ counterparts (Figure 6D). In summary, data presented in Figure 6 showed that FOXO3 deficiency increased the quantity of CD8 T cell memory without affecting the quality. It is well established that memory CD8 T cells are maintained for extended periods of time by proliferative renewal, driven by homeostatic cytokines IL-7 and IL-15 [11], [56], [66]–[68]. Although it is known that IL-7 and IL-15 signaling triggers phosphorylation of FOXO3 [56], [62], the effect of FOXO3 deficiency on proliferative renewal of memory CD8 T cells has not been examined. Using three approaches, we compared the cytokine-driven proliferative renewal of memory CD8 T cells in +/+ and FOXO3−/− mice. First, in vivo BrdU incorporation studies showed that the percentages of BrdU+ve LCMV-specific memory CD8 T cells in FOXO3−/− mice were comparable to those in +/+ mice (Figure 7A). Likewise, the percentages of Ki67+ve LCMV-specific CD8 T cells were unaffected by FOXO3 deficiency (Figure 7A). To further examine the effect of FOXO3 deficiency on the proliferative renewal of memory CD8 T cells, CD8 T cells from the spleens of LCMV-immune +/+ and FOXO3−/− mice were labeled with CFSE and adoptively transferred into congenic uninfected mice. Thirty days after cell transfer, flow cytometric analysis of CFSE staining revealed that donor LCMV-specific CD8 T cells from +/+ and FOXO3−/− mice proliferated equally in the recipient mice (Figure 7B). Taken together, data in Figure 7 provided strong evidence that FOXO3 deficiency did not alter the homeostatic turnover of LCMV-specific memory CD8 T cells. To observe the effect of FOXO3 deletion on recall responses of memory CD8 T cells and protective immunity, LCMV-immune +/+ and FOXO3−/− mice were challenged with LCMV clone 13, a strain of LCMV that establishes a chronic infection in naïve immunocompetent mice. At day 5 after challenge, the numbers of NP396- (P<0. 01), GP33- (P<0. 01) and GP276- (P<0. 03) specific CD8 T cells in FOXO3−/− mice (Figure 8A) were significantly higher than in +/+ mice. The increased number of LCMV-specific CD8 T cells in spleens of LCMV clone 13-Challenged FOXO3−/− mice correlated with the increased numbers of memory CD8 T cells (Figure 6A). As in a primary infection (Figure 1B), we did not see a statistically significant increase in the numbers of LCMV-specific CD8 T cells in either the liver or lymph nodes (Figure 8A). Staining for Ki67 illustrated that FOXO3 deficiency did not affect the proliferation of LCMV-specific CD8 T cells during a secondary response (Figure 8B). Secondary effector CD8 T cells in spleens of FOXO3−/− mice produced comparable levels of IFNγ, TNFα, and IL-2 (Figure 8C). LCMV titers in tissues were comparable in +/+ and FOXO3−/− mice, which indicated that protective immunity was not compromised in the absence of FOXO3 (Figure 8D). To address whether FOXO3 has a T cell intrinsic role in regulating polyclonal CD8 T cell responses to LCMV, we used a cre-loxP knockout strategy to generate the FOXO3L mice that lacked FOXO3 only in T cells [69], [70]. T cell-specific loss of FOXO3 in FOXO3L mice was confirmed by western blot and flow cytometry (Figure S3). FOXO3L and littermate +/+ mice were infected with LCMV and virus-specific CD8 T cell responses were analyzed at day 8 PI. In the spleen, FOXO3L mice exhibited a statistically significant increase in the numbers of LCMV-specific CD8 T cells (NP396 P<0. 02; GP33 P<0. 01; GP276 P<0. 04) over their +/+ littermate controls (Figure 9A). It should be noted that the observed increase in the expansion of CD8 T cells in global FOXO3−/− mice (Figure 1B) was fully recapitulated in FOXO3L mice (Figure 9A). To assess whether greater accumulation of effector CD8 T cells in spleens of FOXO3L mice was driven by increased proliferation, we measured Ki67 expression and BrdU incorporation during the clonal expansion phase of the CD8 T cell response to LCMV. At day 6 PI, the percentages of both Ki67+ve and BrdU+ve LCMV-specific CD8 T cells in FOXO3L mice were comparable to those in +/+ mice, which suggested that enhanced accumulation of effector CD8 T cells in FOXO3L mice are not linked to an altered proliferation rate (Figure 9B). Likewise, percentages of Ki67+ve CD8 T cells at day 5 PI were comparable in +/+ and FOXO3L mice (Figure S4). However, the percentages of Annexin VHI LCMV-specific CD8 T cells in spleens of FOXO3L mice were significantly lower than in +/+ mice (Figure 9B). These data suggested that FOXO3 controls the accumulation of CD8 T cells during a primary response by regulating apoptosis of proliferating cells. Conditional deficiency of FOXO3 in T cells did not affect the cell surface phenotype (data not shown) or antigen-triggered cytokine production by LCMV-specific effector CD8 T cells at day 8 PI (Figure 9C). Since both CD8 and CD4 T cells lack FOXO3 activity in FOXO3L mice, it could be argued that increased clonal expansion of CD8 T cells might result from enhanced CD4 T cell help in FOXO3L−/− mice. To address this issue, we depleted CD4 T cells in +/+ and FOXO3L mice and quantified CD8 T cell responses to LCMV in the absence of CD4 T cells. At day 8 PI, >95% of the CD4 T cells were depleted in spleen of both +/+ and FOXO3L mice (data not shown). The number of LCMV-specific CD8 T cells in spleen of CD4-depleted FOXO3L mice was substantially higher than in CD4 T cell-depleted +/+ mice (Figure 9D). Thus, in the apparent absence of CD4 T cells, FOXO3 deficiency in CD8 T cells was sufficient to increase the accumulation of virus-specific CD8 T cells during an acute LCMV infection. To determine whether deletion of FOXO3, exclusively from the T cell compartment, would affect CD8 T cell memory generation, FOXO3L and +/+ mice were infected with LCMV and virus-specific memory CD8 T cells were quantified in lymphoid and non-lymphoid tissues at 180 days PI. We observed a significant increase in the number of memory CD8 T cells that are specific to the three immuno-dominant LCMV epitopes (NP396 P<0. 02; GP33 P<0. 01; GP276 P<0. 05) in spleens of FOXO3L mice over the +/+ mice (Figure 10A). These data indicated that FOXO3 regulates the magnitude of CD8 T cell memory by T cell intrinsic mechanisms. Next, we examined whether conditional deficiency of FOXO3 in T cells affected the phenotypic and functional attributes of memory CD8 T cells. The expressions of CD44, CD62L, CCR7 and LFA-1 on +/+ and FOXO3L memory CD8 T cells from spleen, liver and lymph nodes were similar (Figure 10B). Additionally, the relative proportions of effector (CD62LLo) and central (CD62LHI) memory CD8 T cells were unaffected by FOXO3 deficiency (Figure 10B). In response to antigenic stimulation, virus-specific memory CD8 T cells in LCMV-immune FOXO3L−/− mice produced IFNγ, TNFα, and IL-2 at levels comparable to those in +/+ mice (Figure 10C). As was the case in our studies of global FOXO3-deficient mice (Figure 7), memory CD8 T cells in FOXO3L mice exhibited no difference in homeostatic turnover, as evidenced by BrdU incorporation from day 120 PI mice, pulsed for 8 days and by parallel Ki67 staining (Figure 10D). Taken together, data presented in Figure 10 strongly suggested that FOXO3 regulates the quantity, with no apparent loss in quality, of CD8 T cell memory by T cell intrinsic mechanisms. The FOXO transcription factors are important regulators of cell cycle progression, apoptosis, and energy metabolism [21], [23], [26], [28], [56]. In the T cell compartment, FOXOs have been implicated in regulating homing of T cells, cytokine receptor expression, and development of regulatory T cells [48], [51], [53], [54]. While it has been reported that FOXO3 may control CD8 T cell expansion, albeit through non T cell intrinsic mechanisms, a role for FOXO3 in memory T cell survival has also been posited [49]. What has not been thoroughly addressed, however, is the T cell intrinsic role of FOXO3 in governing different facets of the physiological polyclonal T cell response to foreign antigens, including the in vivo generation and maintenance of CD8 T cell memory. In the present study, we have systematically examined the T cell-intrinsic role of FOXO3 in controlling the expansion, contraction, and memory phases of the polyclonal CD8 T cell response to an acute viral infection. These studies have provided strong evidence supporting a T cell intrinsic role for FOXO3 in limiting the magnitude of expansion and the number of memory CD8 T cells in a tissue-specific fashion during a physiological response to an acute LCMV infection. These findings have advanced our mechanistic understanding of CD8 T cell homeostasis, and are expected to have implications in the development of effective vaccines. FOXOs are known to maintain cellular quiescence by mechanisms including the induction of cell cycle inhibitors like p27KIP1 and p21Cip1 [30], [34], [37], [38]. Downregulation of FOXO activity is believed to be an obligatory step for cell cycle entry in response to mitogenic stimuli [30]. The observed phosphorylation of FOXO1/O3 in LCMV-specific CD8 T cells was readily detectable at day 5 PI but exhibited a sharp decline by day 8 PI. The drop in the phosphorylation in FOXO1/O3 between days 5 and 8 PI coincides with declining viral load and decreased antigenic stimulation. However, we observed a rebound in the phosphorylation of FOXO1/O3 between days 8 and 11 PI. What controls the dynamics of FOXO1/O3 phosphorylation during a CD8 T cell response? In addition to TCR signaling, FOXO1/O3 phosphorylation is regulated by signaling via cytokine receptors such as IL-2, IL-7, and IL-15 [38], [56], [66]. It is possible that FOXO1/O3 is phosphorylated by different extracellular signals at different phases of the T cell response. For example, during the phase of antigen-driven proliferation, IL-7R expression is known to be very low [68], and TCR signaling along with IL-2/IL-15 might drive the phosphorylation of FOXO1/O3. However, after antigen clearance, IL-7 signaling might drive phosphorylation on the surviving IL-7 receptor-expressing MPECs, and eventually their resultant memory cells. FOXO3 has been reported to suppress expansion of CD8 T cells indirectly by inhibiting IL-6 production by dendritic cells [49]. In this report, however, the T cell intrinsic role of FOXO3 was not assessed in polyclonal CD8 T cells, and monoclonal TCR transgenic CD8 T cells may not always mimic the responses of polyclonal CD8 T cells. In the present study, global FOXO3-deficient mice exhibit increased expansion of polyclonal CD8 T cells specific to multiple epitopes during an acute LCMV infection. Furthermore, infection of T cell-specific conditional FOXO3-deficient mice with LCMV, fully recapitulated the enhanced CD8 T cell expansion seen in global FOXO3−/− mice; even in the absence of CD4 T cells. Therefore, our data implies that FOXO3 suppresses CD8 T cell expansion in vivo by T cell intrinsic and extrinsic mechanisms. This inference is also supported by the reported T cell intrinsic regulation of regulatory T cell development by FOXO1 and FOXO3 [71], [72]. One of the most interesting findings presented in this manuscript is that the effect of FOXO3 deficiency on CD8 T cell expansion and memory is observed in the spleen, but not in the liver or lymph nodes. The enhanced accumulation of effector CD8 T cells preferentially in spleens of FOXO3−/− mice could not be explained by tissue-specific differences in BIM expression in CD8 T cells directly ex vivo; regardless of the tissue (spleen, liver, or lymph nodes), BIM levels in FOXO3−/− CD8 T cells were slightly lower than in +/+ CD8 T cells (Figure S5). Additionally, the selective increase in the number of memory CD8 T cells in spleens of FOXO3−/− mice could not be linked to alterations in the expression of molecules such as CD62L, LFA-1, CCR7, and CD44 that regulate T cell trafficking (Figure 10B). Tissue-specific effects were observed in both global and T cell-specific conditional FOXO3 knockout mice, which suggests that the local immunological milieu influences the effects of FOXO3 in T cells. It is possible that FOXO3-deficient T cells are hyper-responsive to cellular and environmental cues unique to the spleen during or after cessation of antigen-driven proliferation. Tissue-specific alterations in CD8 T cell homeostasis are not unique to FOXO3 deficiency because lymph node-specific effects on CD8 T cell numbers have been reported in mice deficient for Fas and BIM [64]. Future experiments will address the mechanisms underlying the tissue-specific effects of FOXO3 in regulating CD8 T cell homeostasis. FOXO3 is known to regulate both proliferation and apoptosis by controlling the transcription of genes like p27Kip1, p130, BIM, and Fas ligand [26], [34], [35], [63], [64]. Therefore, the observed increase in the accumulation of LCMV-specific CD8 T cells in FOXO3−/− mice could be attributed to altered proliferation and/or apoptosis. Analysis of in vivo proliferation by multiple strategies indicated that FOXO3 deficiency did not alter proliferation rates of LCMV-specific CD8 T cells in vivo. Clonal expansion of CD8 T cells is associated with concomitant proliferation and apoptosis, therefore, cellular accumulation is the result of the proliferation rate exceeding the apoptotic rate. The competing effects of TGF-β and IL-15 are known to dictate the apoptotic rate of proliferating CD8 T cells, but the signaling mechanisms involved are not well defined [61]. We theorized that IL-7/IL-15 deprivation during antigen-driven proliferation might diminish FOXO3 phosphorylation, and augment the expression of BIM. We find that at day 6 PI, apoptosis of LCMV-specific CD8 T cells was significantly reduced in spleens of FOXO3−/− mice. Additionally, IL-15 deprivation was indeed associated with higher BIM levels in +/+ CD8 T cells than in FOXO3−/− CD8 T cells, which suggested that proliferating FOXO3−/− CD8 T cells might be less susceptible to cytokine withdrawal-induced apoptosis during the expansion phase of the CD8 T cell response. During the early contraction phase (day 8–11 PI), a substantial number of LCMV-specific CD8 T cells are still in cycle (Figure 5A), but during this interval the apoptotic rate presumably exceeds the proliferation rate resulting in a net loss of CD8 T cells. Interestingly, FOXO3 deficiency minimally altered the contraction of LCMV-specific CD8 T cells. These data suggest that mechanisms controlling apoptosis of CD8 T cells during expansion and contraction are likely distinct. Remarkably, the numbers of memory CD8 T cells in the spleen of both FOXO3−/− and FOXO3L mice were substantially higher than in +/+ mice. The number of memory CD8 T cells is dictated by the magnitude of expansion (clonal burst size) and contraction of effector CD8 T cells [5]. The magnitude of increase in the number of memory CD8 T cells in FOXO3−/− or FOXO3L mice reflects enhanced expansion of MPECs (Figure 2B) during the primary CD8 T cell response. Importantly, enhancement in the number of memory CD8 T cells induced by FOXO3 deficiency was not associated with detectable alterations in phenotype or effector function. Memory CD8 T cells in LCMV-immune FOXO3−/− mice exhibit strong recall responses and provide effective immunity against a persistent LCMV infection. Thus, FOXO3 deficiency increased the quantity of CD8 T cell memory without affecting their phenotype or effector functions. Memory CD8 T cells are maintained by IL-7 and IL-15-driven proliferative renewal and phosphorylation of FOXO3 is an integral component of the signaling circuitry triggered by IL-7/IL-15 signaling [11], [54], [66], [67]. Additionally, we have previously shown that deficiency of the cell cycle inhibitor p27Kip1, a target gene for FOXO3, enhances the homeostatic turnover of memory CD8 T cells [43]. Surprisingly, despite the suggested importance of FOXO3 in regulating the homeostasis of memory T cells, FOXO3 deficiency exerted minimal effects on the proliferative renewal of antigen-specific memory CD8 T cells in vivo [48]. Studies of human memory T cells have ascribed a negative regulatory role for FOXO3 in the persistence of memory CD4 T cells and FOXO3 deficiency would be expected to increase the number of effector memory cells [48], [73]. However, FOXO3 deficiency did not affect the relative proportions of central and effector memory CD8 T cells. It is plausible that FOXO3 might regulate the persistence of central/effector memory CD4 T cells, and not CD8 T cells. Alternatively, FOXO3 function may be redundant in maintaining fully differentiated memory CD4 and CD8 T cells. In conclusion, this manuscript documents that FOXO3 plays a critical role in controlling the clonal burst size and the magnitude of CD8 T cell memory by T cell intrinsic mechanisms. Furthermore, the enhanced number of memory CD8 T cells induced by FOXO3 deficiency, is maintained for extended periods without compromising its quality. These findings have important implications in vaccine development, and suggest that modulation of FOXO3 activity during the expansion phase might be a fruitful strategy to bolster vaccine-induced CD8 T cell memory and protective immunity. The generation and characterization of the global FOXO3-deficient (FOXO3−/−) mice on the C57BL/6 (B6) background have been described previously [40]. The control wild type B6 (+/+) mice were either littermates or purchased from the National Cancer Institute (Bethesda, MD). Derivation of mice carrying the floxed FOXO3 alleles has been described elsewhere [69], [70]. Mice carrying the floxed FOXO3 alleles were bred with the CD4-Cre mice at UW-Madison to generate the T cell-specific FOXO3−/− (FOXO3L) mice. Littermate +/+ mice were used as controls with the FOXO3L mice. Mice used in experiments were between the ages of 6–8 weeks and all experiments were performed in accordance with the protocols approved by the University of Wisconsin School of Veterinary Medicine Institutional Animal Care and Use Committee (IACUC). The animal committee mandates that institutions and individuals using animals for research, teaching, and/or testing much acknowledge and accept both legal and ethical responsibility for the animals under their care, as specified in the Animal Welfare Act (AWA) and associated Animal Welfare Regulations (AWRs) and Public Health Service (PHS) Policy. Mice were infected with 2×105 PFU of lymphocytic choriomeningitis virus (LCMV) Armstrong strain by intraperitoneal (IP) injection. Mice that have recovered from an infection with LCMV Armstrong were challenged with LCMV-Clone 13 (2×106 PFU by intravenous injection). Tissue viral titers were quantified by plaque assay with Vero cell monolayers [74]. Single cell suspensions of splenocytes were stained with antibodies for surface markers including CD8, CD44, CD122, CD127, CD62L, CCR7, LFA-1 and KLRG-1 (BD Biosciences, Franklin Lakes NJ, eBIOSCEINCE, San Diego CA or Southern Biotech, Birmingham AL) in conjunction with MHC I tetramers (Db) specific for the class I-restricted LCMV epitopes, NP396, GP33, and GP276 as previously described [15]. Cells were fixed in 2% paraformaldehyde (PFA) and acquired in a FACSCalibur or LSR II flow cytometer (BD Biosciences, Franklin Lakes NJ). To quantify intracellular cytokine production, splenocytes were incubated for 5 hours at 37°C with LCMV epitope viral peptides in the presence of Brefeldin A. After stimulation, cells were first incubated with antibodies for surface markers. Next, cells were permeabilized and stained for intracellular cytokines (IFNγ, IL-2 and TNFα) using the Cytofix/Cytoperm kit (BD Biosciences, Franklin Lakes NJ). The percentages of cytokine-producing cells were quantified by flow cytometry. Splenocytes were stained for cell surface markers as above. After cell surface staining, cells were fixed and permeabilized using Phosflow lysis and Phosflow PermWash I reagents (BD Biosciences, Franklin Lakes NJ) according to the manufacturer' s recommendations. Next, cells were blocked for 30 minutes on ice in blocking buffer (10% normal goat serum in 2%BSA/PBS) and subsequently stained with either phospho-specific antibodies (Cell Signaling Technology, Danvers MA; P-Akt [T308], P-FOXO1/3 [T24, T32], P-mTOR [S2448]) or non-phospho state-specific antibodies (Akt, mTOR, FOXO3, BIM). As negative controls for staining, antibodies were pre-incubated/blocked with their specific antigenic peptide for 1 hr at room temperature before adding on to the cells. Following incubation with antibody or peptide blocked antibody, cells were washed twice and incubated with secondary antibody (Goat anti-Rabbit ALEXA488; Sigma-Aldrich, St. Louis MO) for 40 minutes. Cells were washed and fixed with 2% PFA. The levels of phospho-specific staining were quantified by flow cytometry. Specific levels of staining (Corrected Mean Fluorescence Intensity [MFI]) were calculated using the formula: difference of observed MFI for the phospho-specific protein and peptide-blocked control, divided by the peptide-blocked control. To assess in vivo proliferation of antigen specific cells, mice were administered an IP injection of 1. 5 mg of 5-Bromo-2′-deoxyuridine ([BrdU] MP Biomedicals, Solon OH) followed by exposure to 0. 8 mg/ml of BrdU in drinking water for the rest of the pulse period. Splenocytes were stained for surface markers and determination of BrdU positive cells was performed using a BrdU staining kit (BD Biosciences). Splenocytes were stained for surface markers and MHC I tetramers as described above. After surface staining, cells were fixed and permeabilized using FACS Lysing Solution and FACS Permeabilization Solution 2 reagents (BD Biosciences, Franklin Lakes NJ) and subsequently incubated with antibodies against Ki67, Bcl-2 or Granzyme B (BD Biosciences, Franklin Lakes NJ) for 45 minutes at room temperature. Virus-specific CD8 T cells staining positive for Ki67, Bcl-2 or Granzyme B were visualized using a FACSCalibur flow cytometer. Data is expressed as either a percentage of antigen-specific CD8 T cells positive for the respective protein or MFI for the indicated protein. After 6 days PI, splenocytes from +/+ and FOXO3−/− or FOXO3L mice were isolated and stained with anti-CD8 and MHC-I tetramers as described above except no red blood cell lysis was performed. Annexin V staining (BD Biosciences, Franklin Lakes NJ) was then carried out according to the manufacturer' s protocol, with the exception that all staining was performed on ice. The percentage of Annexin V high cells amongst antigen-specific CD8 T cells was determined by flow cytometry and expressed accordingly. Mice were depleted of CD4 T cells through IP administration of 100 µg of the monoclonal antibody GK1. 5 (eBioscience, San Diego CA) at days 0 and 4 relative to LCMV infection. T cells and non-T cells were purified from spleens of +/+ and FOXO3L mice using the anti-CD90. 2 based MACS cell separation system (Miltenyi Biotec, Auburn CA). Purity of cells was >93%. Cells were subsequently lysed in buffer (50 mM HEPES, 100 Mm NaCl, 10 mM EDTA, 10 Mm NaF, 4 Mm Na (PO4) 2,1% Triton X-100,5 µg/ml Aprotinin, 1 Mm Phenylmethylsulfonylflouride), sonicated, and total protein levels in each lysate were determined by the Bicinchoninic Acid protein assay. (Sigma-Aldrich, St. Louis, MO). 20 µg samples were loaded and resolved on a 10% SDS-PAGE. Total levels of FOXO3 protein in each sample were detected using a Rabbit primary antibody specific for FOXO3 (Cell Signaling Technology, Danvers MA) followed by a Donkey anti Rabbit F (ab) 2 fragment HRP-conjugated secondary antibody (Thermo Fisher, Rockford IL) Bands were visualized using chemiluminescence reagents (Thermo Fisher, Rockford IL) and presented by use of an HP Deskscan system (Hewlett-Packard, Palo Alto CA). Blots initially probed for FOXO3 were subsequently stripped and re-probed with β-Actin (Sigma-Aldrich, St. Louis MO) to serve as a loading control.

Title: FOXO3 Regulates CD8 T Cell Memory by T Cell-Intrinsic Mechanisms Summary: CD8 T cells are vital for controlling infections with viruses, intracellular bacteria and protozoa. Induction of T and B cell memory is the basis of vaccinations and cellular immunity to intracellular pathogens depends upon the number and quality of memory CD8 T cells. Understanding the mechanisms that control various facets of CD8 T cell memory is of fundamental importance for development of effective vaccines. In this study, we have identified the transcription factor FOXO3 as a crucial regulator of the magnitude of CD8 T cell memory. During a T cell response, FOXO3 limits the number of memory CD8 T cells by inhibiting the accumulation of memory precursor effector cells that give raise to long-lived CD8 T cells. Loss of FOXO3 activity in T cells led to a durable increase in the number of memory CD8 T cells, and the functional quality of FOXO3-deficient memory CD8 T cells was unaffected by FOXO3 deficiency. Thus, our studies suggest that targeting FOXO3 activity may be a fruitful strategy to augment vaccine-induced CD8 T cell memory and protective immunity.

lay_plos

en

Summarize: WESTMINSTER, Colo. — Vincent Nett, the student who set himself on fire in the cafeteria of Standley Lake High School last month, died of his injuries Sunday evening, the Westminster Police Department confirmed. Nett, a 16-year-old sophomore, used a one-gallon can of Coleman fuel to burn himself. Officials do not believe he intended to injure anyone else in the incident. “We do believe that this was a suicide attempt we don’t have any indication at this point that there was a threat against Standley Lake High School or any other school in Westminster,” Westminster Police spokeswoman Cheryl Spottke said. “I saw this explosion happening in the lunch room,” student Leif Samson said. ”Everyone started running out screaming, ‘Fire!’ I just grabbed my backpack and ran upstairs, and I looked out the windows overlooking the cafeteria. That’s when I found out it was a guy on fire.” Two employees used a fire extinguisher to put out the fire quickly, but not before Nett suffered severe burns. One of the first emergency responders estimated he had third degree burns over 80 percent of his body. “This is just such a tragedy,” Jefferson County Schools spokesperson Lynn Setzer said. “Our hearts and our prayers go out to the family and to the school community and to the students who were here (that) morning.” Investigators later searched Nett’s home, where they learned about a suicide note posted on Facebook that claims he had been planning his own public death for years. Local News Police cars block the entrance to Standley Lake High School in Westminster after a fire in the cafeteria on Jan. 27, 2014. (Kathryn Osler, The Denver Post) A 16-year-old Westminster boy died Sunday afternoon almost two weeks after he set himself on fire in the cafeteria at Standley Lake High School, the Westminster Police Department said. Vincent Nett was burned over 80 percent of his body, and about 40 percent of the burns were third-degree, a Westminster Fire Department spokeswoman said after the Jan. 27 incident. He died just before 5 p.m. Sunday, Westminster police said. Witnesses said Nett entered the cafeteria at 7:12 a.m. carrying a 1-gallon Coleman fuel jug. When he set himself on fire, there were more than 60 students present. None of the other students were injured, but a school worker was cut breaking glass to retrieve a fire extinguisher. Nett posted a suicide note on his Facebook page. "This is not someone's fault, I had this planned for years so shut your face if you think that this was because of something recent or because of someone," he wrote. "There was nothing that anyone could have done." Counselors will be available to students at the school Monday, the police department said. Joey Bunch: 303-954-1174, jbunch@denverpost.com or twitter.com/joeybunch

Summary: The 16-year-old who set himself on fire in his Colorado high school's cafeteria late last month died yesterday, the Denver Post reports. Vincent Nett suffered burns over 80% of his body in the Jan. 27 incident, with 40% of those burns being third-degree. "This is not someone's fault, I had this planned for years so shut your face if you think that this was because of something recent or because of someone," Vincent had written in a suicide note before walking into the Standley Lake High School cafeteria with a 1-gallon Coleman fuel jug shortly after 7am. Police don't think he intended to harm anyone else, KDVR reports.

multi_news

en

Write a title and summarize: SECTION 1. SHORT TITLE. This Act may be cited as the ``Defend and Save Social Security Act''. SEC. 2. ADJUSTMENT TO NORMAL AND EARLY RETIREMENT AGE. (a) In General.--Section 216(l) of the Social Security Act (42 U.S.C. 416(l)) is amended-- (1) in paragraph (1)-- (A) in subparagraph (C), by striking ``2017'' and inserting ``2016''; and (B) by striking subparagraphs (D) and (E) and inserting the following new subparagraphs: ``(D) with respect to an individual who-- ``(i) attains 62 years of age after December 31, 2015, and before January 1, 2024, such individual's early retirement age (as determined under paragraph (2)(A)) plus 48 months; or ``(ii) receives a benefit described in paragraph (2)(B) and attains 60 years of age after December 31, 2015, and before January 1, 2024, 66 years of age plus the number of months in the age increase factor (as determined under paragraph (4)(A)(i)); ``(E) with respect to an individual who-- ``(i) attains 62 years of age after December 31, 2023, and before January 1, 2027, 68 years of age plus the number of months in the age increase factor (as determined under paragraph (4)(B)(ii)); or ``(ii) receives a benefit described in paragraph (2)(B) and attains 60 years of age after December 31, 2023, and before January 1, 2027, 68 years of age plus the number of months in the age increase factor (as determined under paragraph (4)(B)(i)); and ``(F) with respect to an individual who-- ``(i) attains 62 years of age after December 31, 2026, 69 years of age; or ``(ii) receives a benefit described in paragraph (2)(B) and attains 60 years of age after December 31, 2026, 69 years of age.''; (2) by amending paragraph (2) to read as follows: ``(2) The term `early retirement age' means-- ``(A) in the case of an old-age, wife's, or husband's insurance benefit-- ``(i) 62 years of age with respect to an individual who attains such age before January 1, 2016; ``(ii) with respect to an individual who attains 62 years of age after December 31, 2015, and before January 1, 2023, 62 years of age plus the number of months in the age increase factor (as determined under paragraph (4)(A)(ii)) for the calendar year in which such individual attains 62 years of age; and ``(iii) with respect to an individual who attains age 62 after December 31, 2022, 64 years of age; or ``(B) in the case of a widow's or widower's insurance benefit, 60 years of age.''; (3) by striking paragraph (3) and inserting the following: ``(3) With respect to an individual who attains early retirement age in the 5-year period consisting of the calendar years 2000 through 2004, the age increase factor shall be equal to two-twelfths of the number of months in the period beginning with January 2000 and ending with December of the year in which the individual attains early retirement age.''; and (4) by adding at the end the following new paragraph: ``(4) The age increase factor shall be equal to three- twelfths of the number of months in the period-- ``(A) beginning with January 2016 and ending with December of the year in which-- ``(i) for purposes of paragraphs (1)(D)(ii), the individual attains 60 years of age; or ``(ii) for purposes of paragraph (2)(A)(ii), the individual attains 62 years of age; and ``(B) beginning with January 2024 and ending with December of the year in which-- ``(i) for purposes of (1)(E)(ii), the individual attains 60 years of age; or ``(ii) for purposes of (1)(E)(i), the individual attains 62 years of age.''. (b) Conforming Increase in Number of Elapsed Years for Purposes of Determining Primary Insurance Amount.--Section 215(b)(2)(B)(iii) of such Act (42 U.S.C. 415(b)(2)(B)(iii)) is amended by striking ``age 62'' and inserting ``early retirement age (or, in the case of an individual who receives a benefit described in section 216(l)(2)(B), 62 years of age)''. SEC. 3. COST-OF-LIVING ADJUSTMENT. Section 215(i) of the Social Security Act (42 U.S.C. 415(i)) is amended-- (1) in paragraph (1)(D), by inserting ``subject to paragraph (6),'' before ``the term''; and (2) by adding at the end the following new paragraph: ``(6)(A) Subject to subparagraph (B), with respect to a base quarter or cost-of-living computation quarter in any calendar year after 2010, the term `CPI increase percentage' means the percentage determined under paragraph (1)(D) for the quarter reduced (but not below zero) by 1 percentage point. ``(B) The reduction under subparagraph (A) shall apply only for purposes of determining the amount of benefits under this title and not for purposes of determining the amount of, or any increases in, benefits under other provisions of law which operate by reference to increases in benefits under this title.''.

Title: A bill to amend title II of the Social Security Act to extend the solvency of the Social Security Trust Funds by increasing the normal and early retirement ages under the Social Security program and modifying the cost-of-living adjustments in benefits Summary: Defend and Save Social Security Act - Amends title II (Old Age, Survivors and Disability Insurance) (OASDI) of the Social Security Act to: (1) increase the normal retirement age by specified graduated stages to 67 by 2019 and to 69 after December 31, 2026, and the early retirement age to 63 by 2019 and to 64 after December 31, 2022; (2) revise requirements for computation of the age increase factor; and (3) modify the cost-of-living adjustment (COLA) to 1% below the general COLA.

billsum

en

Summarize: Background In May 1997, we reported on DOD’s actions to improve deployment health surveillance before, during, and after deployments, focusing on Operation Joint Endeavor, which was conducted in the countries of Bosnia-Herzegovina, Croatia, and Hungary. We commented on the provisions of a joint medical surveillance policy draft that called for a comprehensive DOD-wide medical surveillance capability to monitor and assess the effects of deployments on servicemembers’ health. DOD subsequently finalized its joint medical surveillance policy in August 1997. Our 1997 review disclosed problems with the Army’s implementation of the medical surveillance plan for Operation Joint Endeavor in the following areas: Medical assessments. Many Army personnel who should have received post-deployment medical assessments did not receive them and the assessments that were completed were frequently done late. Of the 618 servicemembers in the 12 Army units whose medical records we reviewed, 24 percent did not receive in-theater post-deployment medical assessments, and 21 percent did not receive home station post-deployment medical assessments. Servicemembers who received home station post-deployment medical assessments received them, on average, nearly 100 days after they left theater instead of within 30 days as required by the plan. Further, pre-deployment blood serum samples were not available for 9.3 percent of the 26,621 servicemembers who had deployed to Bosnia as of March 12, 1996. The most recent blood samples for 6.4 percent of the pre-deployment blood samples were more than 5 years old. Medical record keeping. Many of the servicemembers’ medical records that we reviewed were incomplete and missing documentation of in-theater post-deployment medical assessments, medical visits during deployment, and receipt of an investigational new vaccine. More specifically, we found that 91 of the 473 servicemembers (19 percent) with a post-deployment in-theater medical assessment and 9 of the 491 servicemembers (1.8 percent) with a post-deployment home unit medical assessment did not have the assessments documented in their medical records. Furthermore, about 29 percent of the 50 battalion aid station visits we reviewed were not documented in the members’ permanent medical records. Finally, 141 of 588 servicemembers (24 percent) who received an investigational drug vaccine did not have the immunization documented in their medical records. Centralized database. The centralized database for collecting in-theater and home unit post-deployment medical assessments was incomplete for many Army personnel. More specifically, the database omitted 12 percent of the in-theater medical assessments done and 52 percent of the home unit medical assessments done for the 618 servicemembers whose records we reviewed. Deployment information. DOD officials considered the database used for tracking the deployment of Air Force and Navy personnel inaccurate. “(a) SYSTEM REQUIRED—The Secretary of Defense shall establish a system to assess the medical condition of members of the armed forces (including members of the reserve components) who are deployed outside the United States or its territories or possessions as part of a contingency operation (including a humanitarian operation, peacekeeping operation, or similar operation) or combat operation. “(b) ELEMENTS OF SYSTEM—The system described in subsection (a) shall include the use of predeployment medical examinations and postdeployment medical examinations (including an assessment of mental health and the drawing of blood samples) to accurately record the medical condition of members before their deployment and any changes in their medical condition during the course of their deployment. The postdeployment examination shall be conducted when the member is redeployed or otherwise leaves an area in which the system is in operation (or as soon as possible thereafter). “(c) RECORDKEEPING—The results of all medical examinations conducted under the system, records of all health care services (including immunizations) received by members described in subsection (a) in anticipation of their deployment or during the course of their deployment, and records of events occurring in the deployment area that may affect the health of such members shall be retained and maintained in a centralized location to improve future access to the records. “(d) QUALITY ASSURANCE—The Secretary of Defense shall establish a quality assurance program to evaluate the success of the system in ensuring that members described in subsection (a) receive predeployment medical examinations and postdeployment medical examinations and that the recordkeeping requirements with respect to the system are met.” As set forth above, these provisions require the use of pre-deployment and post-deployment medical examinations to accurately record the medical condition of servicemembers before deployment and any changes during their deployment. In a June 30, 2003, correspondence with the General Accounting Office, the Assistant Secretary of Defense for Health Affairs stated that “it would be logistically impossible to conduct a complete physical examination on all personnel immediately prior to deployment and still deploy them in a timely manner.” Therefore, DOD required both pre- and post-deployment health assessments for servicemembers who deploy for 30 or more continuous days to a land-based location outside the United States without a permanent U.S. military treatment facility. Both assessments use a questionnaire designed to help military healthcare providers in identifying health problems and providing needed medical care. The pre-deployment health assessment is generally administered at the home station before deployment, and the post-deployment health assessment is completed either in theater before redeployment to the servicemember’s home unit or shortly upon redeployment. As a component of medical examinations, the statute quoted above also requires that blood samples be drawn before and after a servicemember’s deployment. DOD Instruction 6490.3, August 7, 1997, requires that a pre-deployment blood sample be obtained within 12 months of the servicemember’s deployment. However, it requires the blood samples be drawn upon return from deployment only when directed by the Assistant Secretary of Defense for Health Affairs. According to DOD, the implementation of this requirement was based on its judgment that the Human Immunodeficiency Virus serum sampling taken independent of deployment actions is sufficient to meet both pre- and post-deployment health needs, except that more timely post-deployment sampling may be directed when based on a recognized health threat or exposure. Prior to April 2003, DOD did not require a post-deployment blood sample for servicemembers supporting the OEF and OJG deployments. In April 2003, DOD revised its health surveillance policy for blood samples and post-deployment health assessments. Effective May 22, 2003, the services are required to draw a blood sample from each redeploying servicemember no later than 30 days after arrival at a demobilization site or home station. According to DOD, this requirement for post-deployment blood samples was established in response to an assessment of health threats and national interests associated with current deployments. The department also revised its policy guidance for enhanced post-deployment health assessments to gather more information from deployed servicemembers about events that occurred during a deployment. More specifically, the revised policy requires that a trained health care provider conduct a face-to-face health assessment with each returning servicemember to ascertain (1) the individual’s responses to the health assessment questions on the post-deployment health assessment form; (2) the presence of any mental health or psychosocial issues commonly associated with deployments; (3) any special medications taken during the deployment; and (4) concerns about possible environmental or occupational exposures. The Army and Air Force Did Not Comply with Deployment Health Surveillance Policies for Many Servicemembers The Army and Air Force did not comply with DOD’s force health protection and surveillance requirements for many of the servicemembers in our samples at the selected installations we visited. Specifically, these Army and Air Force servicemembers were missing: pre-deployment and/or post-deployment health assessments; evidence of receiving one or more of the pre-deployment immunizations required for their deployment location; and other pre-deployment requirements related to tuberculosis screening and blood serum sample storage. Also, servicemembers’ permanent medical records were missing required health-related information, and DOD’s centralized database did not include documentation of servicemember health-related information. Neither the installations nor DOD had monitoring and oversight mechanisms in place to help ensure that the force health protection and surveillance requirements were met for all servicemembers. Many Servicemembers Lacked Pre-deployment and Post-deployment Health Assessments We found that servicemembers missing one or both of their pre- and post-deployment assessments ranged from 38 to 98 percent in our samples. For example, at Fort Campbell for the OEF deployment we found that 68 percent of the 222 active duty servicemembers in our sample were missing either one or both of the required pre-deployment and post- deployment health assessments. The results of our statistical samples for the deployments at the installations visited are depicted in figure 1. For those servicemembers in our samples who had completed pre- or post-deployment health assessments, we found that as many as 45 percent of the assessments in our samples were not completed on time in accordance with requirements (see fig. 2). DOD policy requires that servicemembers complete a pre-deployment health assessment form within 30 days of their deployment and a post-deployment health assessment form within 5 days upon redeployment back to their home station. These time frames were established to allow time to identify and resolve any health concerns or problems that may affect the ability of the servicemember to deploy, and to promptly identify and address any health concerns or problems that may have arisen during the servicemember’s deployment. Not all health assessments were reviewed by a health care provider as required, as shown in figure 3. DOD policy requires that pre-deployment and post-deployment health assessments are to be reviewed immediately by a health care provider to identify any medical care needed by the servicemember. The services did not refer some servicemember health assessments that indicated a need for further consultation. According to DOD policy, a medical provider, namely a physician, physician’s assistant, nurse, or independent duty medical technician is required to further review a servicemember’s need for specialty care when the member’s pre-deployment and/or post-deployment health assessment indicates health concerns such as unresolved medical or dental problems or plans to seek mental health counseling or care. This follow-up may take the form of an interview or examination of the servicemember, and forms the basis of a decision as to whether a referral for further specialty care is warranted. In our samples, the number of assessments that indicated a health concern was relatively small, but large percentages of these assessments were not referred for further specialty care. For example, our sample at Travis Air Force Base included five pre-deployment health assessments that indicated a health concern, but four (80 percent) of the health assessments were not referred for further specialty care. Noncompliance with the requirement for pre-deployment health assessments may result in servicemembers with existing health problems or concerns being deployed with unaddressed health problems. Also, failure to complete post-deployment health assessments may risk a delay in obtaining appropriate medical follow-up attention for a health problem or concern that may have arisen during or following the deployment. Immunizations and Other Pre-Deployment Health Requirements Not Met Based on our samples, the services did not fully meet immunization and other pre-deployment requirements. Evidence of pre-deployment immunizations receipt was missing from many servicemembers’ medical records. Servicemembers missing the required immunizations may not have the immunization protection they need to counter theater disease threats. Based on our review of servicemember medical records for the deployments at the four installations we visited, we found that between 14 and 46 percent of the servicemembers were missing at least one of their required immunizations prior to deployment (see fig. 4). Furthermore, as many as 36 percent of the servicemembers were missing two or more of their required immunizations. The U.S. Central Command required the following pre-deployment immunizations for all servicemembers that deployed to Central Asia in support of OEF: hepatitis A (two-shot series); measles, mumps, and rubella; polio; tetanus/diphtheria within the last 10 years; yellow fever within the last 10 years; typhoid within the last 5 years; influenza within the last 12 months; and meningococcal within the last 5 years. For OJG deployments, the U.S. European Command required the same immunizations cited above, with the exception of the yellow fever inoculation that was not required for Kosovo. Figure 5 indicates that 7 to 40 percent of the deploying servicemembers in our review were missing a current tuberculosis screening. A screening is deemed “current” if it occurred 1 to 2 years prior to deployment. Specifically, the U.S. Central Command required servicemembers deploying to Central Asia in support of OEF to be screened for tuberculosis within 12 months of deployment. For OJG deployments, the U.S. European Command required Army and Air Force servicemembers to be screened for tuberculosis with 24 months of deployment. U.S. Central Command and U.S. European Command policies require that deploying servicemembers have a blood serum sample in the serum repository not older than 12 months prior to deployment. While nearly all deploying servicemembers had blood serum samples held in the Armed Services Serum Repository prior to deployment, as many as 29 percent had serum samples that were too old (see table 1). The samples that were too old ranged, on average, from 2 to 15 months out-of-date. Servicemember Medical Records and Centralized Database Were Not Complete Servicemembers’ permanent medical records were not complete, and DOD’s centralized database did not include documentation of servicemember health-related information. Many servicemembers’ permanent medical records at the Army and Air Force installations we visited did not include documentation of completed health assessments and servicemember visits to Army battalion aid stations. Similarly, the centralized deployment record database did not include many of the deployment health assessments and immunization records that we found in the servicemembers’ medical records at the installations we visited. Many Completed Deployment Health Assessments and Medical Interventions Were Not Documented in Servicemembers’ Medical Record DOD policy requires that the original completed pre-deployment and post-deployment health assessment forms be placed in the servicemember’s permanent medical record and that a copy be forwarded to AMSA. Figure 6 shows that completed assessments we found at AMSA and at the U.S. Special Operations Command for servicemembers in our samples were not documented in the servicemember’s permanent medical record, ranging from 8 to 100 percent for pre-deployment health assessments and from 11 to 62 percent for post-deployment health assessments. Army and Air Force policies also require documentation in the servicemember’s permanent medical record of all visits to in-theater medical facilities. Except for the OEF deployment at Fort Drum, officials were unable to locate or access the sign-in logs for servicemember visits to in-theater Army battalion aid stations and to Air Force expeditionary medical support for the OEF and OJG deployments at the installations we visited. Consequently, we limited the scope of our review to two battalion aid stations for the OEF deployment at Fort Drum. We found that 39 percent of servicemember visits to one battalion aid station and 94 percent to the other were not documented in the servicemember’s permanent medical record. Representatives of the two battalion aid stations said that the missing paper forms documenting the servicemember visits may have been lost en route to Fort Drum. Specifically, a physician’s assistant for one of these battalion aid station said the battalion aid station moved three times in theater and each time the paper forms used to document in-theater visits were boxed and moved with the battalion aid station. Consequently, the forms missing from servicemembers’ medical records may have been lost en route to Fort Drum. The lack of complete and accurate medical records documenting all medical care for the individual servicemember complicates the servicemembers’ post-deployment medical care. For example, accurate medical records are essential for the delivery of high-quality medical care and important for epidemiological analysis following deployments. According to DOD health officials, the lack of complete and accurate medical records complicated the diagnosis and treatment of servicemembers who experienced post-deployment health problems that they attributed to their military service in the Persian Gulf in 1990-91. DOD is implementing the Theater Medical Information Program (TMIP) that has the capability to electronically record and store in-theater patient medical encounter data. TMIP is currently undergoing operational testing by the military services and DOD intends to begin fielding TMIP during the first quarter of fiscal year 2004. Centralized Database Missing Health-Related Documentation Based on our samples, DOD’s centralized database did not include documentation of servicemember health-related information. As set forth above, Public Law 105-85, enacted November 1997, requires the Secretary of Defense to retain and maintain health-related records in a centralized location. This includes records for all medical examinations conducted to ascertain the medical condition of servicemembers before deployment and any changes during their deployment, all health care services (including immunizations) received in anticipation of deployment or during the deployment, and events occurring in the deployment area that may affect the health of servicemembers. A February 2002 Joint Staff memorandum requires the services to forward a copy of the completed pre-deployment and post-deployment health assessments to AMSA for centralized retention. Also, the U.S. Special Operations Command (SOCOM) requires deployment health assessments for special forces units to be sent to the Command for centralized retention in the Special Operation Forces Deployment Health Surveillance System. Figure 7 depicts the percentage of pre- and post-deployment health assessments and immunization records we found in the servicemembers’ medical records that were not available in a centralized database at AMSA or SOCOM. Health-related documentation missing from the centralized database ranged from 0 to 63 percent for pre-deployment health assessments, 11 to 75 percent for post-deployment health assessments, and 8 to 93 percent for immunizations. All but one of the servicemembers in our sample at Hurlburt Field were special operations forces. A SOCOM official told us that pre-deployment and post-deployment health assessment forms for servicemembers in special operations force units are not sent to AMSA because the health assessments may include classified information that AMSA is not equipped to receive. Consequently, SOCOM retains the deployment health assessments in its classified Special Operations Forces Deployment Health Surveillance System. Also, a SOCOM medical official told us that the system does not include pre-deployment immunization data. A Deployment Health Support Directorate official told us that the Directorate is examining how to remove the classified information from the deployment health assessments so that SOCOM can forward the assessments to AMSA. For presentation in figure 7, we combined the health assessment and immunization data we found at AMSA and SOCOM for Hurlburt Field. An AMSA official believes that missing documentation in the centralized database could be traced to the services’ use of paper copies of deployment health assessments that installations are required to forward to the centralized database, and the lack of automation to record servicemembers’ pre-deployment immunizations. DOD has ongoing initiatives to electronically automate the deployment health assessment forms and the recording of servicemember immunizations. For example, DOD is implementing a comprehensive electronic medical records system, known as the Composite Health Care System II, which includes pre- and post-deployment health assessment forms and the capability to electronically record immunizations given to servicemembers. DOD has deployed the system at five sites and will be seeking approval in August/September 2003 for worldwide deployment. DOD officials believe that the electronic automation of the deployment health-related information will lessen the burden of installations in forwarding paper copies and the likelihood of information being lost in transit. DOD and Installations Did Not Have Oversight of Force Health Protection and Surveillance Requirements DOD does not have an effective quality assurance program to provide oversight of, and ensure compliance with, the department’s force health protection and surveillance requirements. Moreover, the installations we visited did not have ongoing monitoring or oversight mechanisms to help ensure that force health protection and surveillance requirements were met for all servicemembers. We believe that the lack of such a system was a major cause of the high rate of noncompliance we found at the units we visited. The services are currently developing quality assurance programs designed to ensure that force health protection and surveillance policies are implemented for servicemembers. Although required by Public Law 105-85 to establish a quality assurance program, neither the Assistant Secretary of Defense for Health Affairs nor the offices of the Surgeons General of the Army or Air Force had established oversight mechanisms that would help ensure that force health protection and surveillance requirements were met for all servicemembers. Following our visit to Fort Drum in October 2002, the Army Surgeon General wrote a memorandum in December 2002 to the commanders of the Army Regional Medical Commands that expressed concern related to our sample results at Fort Drum. He emphasized the importance of properly documenting medical care and directed them to accomplish an audit of a statistically significant sample of medical surveillance records of all deployed and redeployed soldiers at installations supported by their regional commands, provide an assessment of compliance, and develop an action plan to improve compliance with the requirements. At three of the four installations we visited, officials told us that new procedures were implemented that they believe will improve compliance with force health protection and surveillance requirements for deployments occurring after those we reviewed. Specifically, following our visit to Fort Drum in October 2002, Fort Drum medical officials designed a pre-deployment and post-deployment checklist patterned after our review that is being used as part of processing before servicemembers are deployed and when they return. The officials told us that this process has improved their compliance with force health protection and surveillance requirements for deployments subsequent to our visit. Also, the hospital commander at Fort Campbell told us that they implemented procedures that now require all units located at Fort Campbell to use the hospital’s medical personnel in their processing of servicemembers prior to deployment. The hospital commander believes that this new requirement will improve compliance with the force health protection and surveillance requirements at Fort Campbell because the medical personnel will now review whether all requirements have been met for the deploying servicemembers. At Hurlburt Field, officials told us that they implemented a new requirement in November 2002 to withhold payment of travel expenses and per diem to re-deploying servicemembers until they complete the post-deployment health assessment. Officials believe that this change will improve servicemembers’ completion of the post-deployment health assessments. While it is noteworthy that these installations have implemented changes that they believe will improve their compliance, the actual measure of improvements over time cannot be known unless the installations perform periodic reviews of servicemembers’ medical records to identify the extent of compliance with deployment health requirements. In March 2003, we briefed the Subcommittee on Total Force, House Committee on Armed Services, about our interim review results at selected military installations. Subsequently, at a March 2003 congressional hearing, the Subcommittee discussed our interim review results with the Assistant Secretary of Defense for Health Affairs and the services’ Surgeons General. Based on our interim results that DOD was not meeting the full requirement of the law and the military services were not effectively carrying out many of DOD’s force health protection and surveillance policies, in May 2003 the House Committee on Armed Services directed the Secretary of Defense to take measures to improve oversight and compliance. Specifically, in its report accompanying the Fiscal Year 2004 National Defense Authorization Act, the Committee directed the Secretary of Defense “… to establish a quality control program to begin assessing implementation of the force health protection and surveillance program, and to provide a strategic implementation plan, including a timeline for full implementation of all policies and programs, to the Senate Committee on Armed Services and the House Committee on Armed Services by March 31, 2004.” In April 2003, the Under Secretary of Defense for Personnel and Readiness issued an enhanced post-deployment health assessment policy that required the services to develop and implement a quality assurance program that encompasses medical record keeping and medical surveillance data. In June 2003, the Office of Assistant Secretary of Defense for Health Affairs’ Deployment Health Support Directorate began reviewing the services’ quality assurance implementation plans and establishing DOD-wide compliance metrics—including parameters for conducting periodic visits—to monitor service implementation. Centralized Deployment Database Still Missing Information Needed for Deployment Health Surveillance The DMDC deployment database still does not include the deployment information we identified in 1997 as needed for effective deployment health surveillance. In 1997, we reported that knowing the identity of servicemembers who were deployed during a given operation and tracking their movements within the theater of operations are major elements of a military medical surveillance system. The Institute of Medicine reported in 2000 that the documentation of the locations of units and individuals during a given deployment is important for epidemiological studies and for the provision of appropriate medical care during and after deployments. This information allows (1) epidemiologists to study the incidence of disease patterns across populations of deployed servicemembers who may have been exposed to diseases and hazards within the theater, and (2) health care professionals to treat their medical problems appropriately. Because of concerns about the accuracy of the DMDC database, we recommended in our 1997 report that the Secretary of Defense direct an investigation of the completeness of the information in the DMDC personnel database and take corrective actions to ensure that the deployment information is accurate for servicemembers who deploy to a theater. DOD’s established policies notwithstanding, the services did not report location-specific deployment information to DMDC prior to April 2003, because, according to a DMDC official, the services did not maintain the data. DOD Instruction 6490.3, issued in August 1997, requires DMDC, under the Department’s Under Secretary for Personnel and Readiness, to maintain a system that collects information on deployed forces, including daily-deployed strength, total and by unit; grid coordinate locations for each unit (company size and larger); and inclusive dates of individual servicemember’s deployment. In addition, the Joint Chief of Staff’s Memorandum MCM-0006-02, dated February 1, 2002, required combatant commands to provide DMDC with their theater-wide rosters of all deployed personnel, their unit assignments, and the unit’s geographic locations while deployed. This memorandum stressed that accurate personnel deployment data is needed to assess the significance of medical diseases and injuries in terms of the rate of occurrence among deployed servicemembers. The Under Secretary of Defense for Personnel and Readiness expressed concern about the services’ failure to report complete personnel deployment data to DMDC in an October 2002 memorandum. To address the services’ lack of reporting to DMDC, the Under Secretary of Defense for Personnel and Readiness established a tri-service working group that outlined a plan of action in March 2003 to address the reporting issues. In July 2003, a DMDC official told us that significant improvements had recently occurred and that all of the services had begun submitting their classified deployment databases—including deployment locations—to DMDC. DMDC is currently reviewing the deployment information submitted by the services to determine its accuracy and completeness. It plans to complete this review during the summer of 2003. With regard to DMDC’s efforts to create a system for tracking the movements of servicemembers within a given theater of operations, DMDC officials told us that little progress has been made. They said that the primary reason for a lack of progress in developing this system is that the source information has generally not been available from the services and this may require the development of new tracking systems at the unit level. In June 2003, a DMDC official told us that it had been recently determined that the Air Force has implemented a theater tracking system that may have applicability to the other services. The tracking system—known as the Deliberate Crisis and Action Planning and Execution Segment (DCAPES)—enables field teams to enter classified information about the whereabouts of deployed Air Force personnel at the longitude/latitude level of detail. DMDC began receiving information from this system in April 2003. The Under Secretary of Defense for Personnel and Readiness is reviewing this system to determine whether it could be used for the same purposes by the other services. Also, DOD is developing the Defense Integrated Military Human Resource System (DIMHRS), which will have the capability to track the movements of all servicemembers and civilians in the theater of operations. As of June 2003, DOD plans to implement this system for the Army by about September 2005 and for the other services by 2007 or early calendar year 2008. Conclusions While DOD and the military services have established force health protection and surveillance policies, at the units we visited we found many instances of noncompliance by the services. Moreover, because DOD and the services do not have an effective quality assurance program in place to help ensure compliance, these problems went undetected and uncorrected. Continued noncompliance with these policies may result in servicemembers with existing health problems or concerns being deployed with unaddressed health problems or without the immunization protection they need to counter theater disease threats. Failure to complete post-deployment health assessments may risk a delay in obtaining appropriate medical follow-up attention for a health problem or concern that may have arisen during or following the deployment. Similarly, incomplete and inaccurate medical records and deployment databases would likely hinder DOD’s ability to investigate the causes of any future health problems that may arise coincident with deployments. Recommendation for Executive Action To improve compliance with DOD’s force health protection and surveillance policies, we recommend that the Secretary of Defense direct the Assistant Secretary of Defense for Health Affairs to establish an effective quality assurance program, as required by section 765 of Public Law 105-85 (10 U.S.C. 1074f), that will ensure that the military services comply with the force health protection and surveillance requirements for all servicemembers. Agency Comments and Our Evaluation The Department of Defense provided written comments on a draft of this report, which are found in appendix II. DOD concurred with the report’s recommendation. The Assistant Secretary of Defense for Health Affairs commented that his office has already established a quality assurance program for pre- and post-deployment health assessments. This program monitors pre- and post-deployment health assessments and blood samples being archived electronically at the Army Medical Surveillance Activity (AMSA) and assures that indicated referrals on the post-deployment health assessments are being conducted by all the services. However, the Assistant Secretary of Defense for Health Affairs’ comments did not discuss how his office is using the monitoring activities to assure the military services’ compliance with force health protection and surveillance policies. According to the Assistant Secretary of Defense for Health Affairs, the services have implemented their quality assurance programs. The Army has developed automated versions of the pre- and post-deployment health assessment forms, and has established a corporate monitoring system that is built upon deployment personnel rosters and monitored weekly by the Army Surgeon General. The Air Force is now receiving monthly deployment health surveillance compliance reports from its medical treatment facilities, and has scheduled a special compliance study through the Air Force Inspection Agency in fiscal year 2004. Navy fleet commanders have implemented their own quality assurance programs, with anticipation of standardization through centralized automated systems. And the Marine Corps has also established unit/command quality assurance procedures. We view these actions as responsive to our recommendation and commend the department for taking quick action to address the compliance issues we found during our audit. However, it remains to be seen how effective these activities will be in ensuring that force health protection and surveillance policies are implemented for all servicemembers. We are sending copies of this report to the Secretary of Defense and the Secretaries of the Army and the Air Force. We will also make copies available to others upon request. In addition, the report is available at no charge on GAO’s Web site at http://www.gao.gov. If you or your staff have any questions regarding this report, please contact me on (757) 552-8100. Key contributors to this report are listed in appendix III. Appendix I: Scope and Methodology To meet our objectives, we interviewed responsible officials and reviewed pertinent documents, reports, and information related to force health protection and deployment health surveillance requirements obtained from officials at the Office of the Assistant Secretary of Defense for Health Affairs; the Office of the Deputy Assistant Secretary of Defense for Force Health Protection and Readiness; the Office of the Assistant Secretary of Defense for Reserve Affairs; the Joint Staff; the Marine Corps Force Health Protection Office; and the Offices of the Surgeons General for the Army and Air Force Headquarters in the Washington, D.C., area. We also performed additional work at the Deployment Health Support Directorate, Falls Church, Virginia; the U.S. Army Center for Health Promotion and Preventive Medicine, Aberdeen, Maryland; the Armed Forces Medical Intelligence Center, Fort Dietrick, Maryland; the Army Medical Surveillance Activity, Walter Reed Army Medical Center, Washington, D.C.; the Navy Environmental Health Center in Portsmouth, Virginia; the Defense Manpower Data Center in Monterey, California; and the U.S. Central Command and the U.S. Special Operations Command at MacDill Air Force Base, Tampa, Florida. To determine whether the military services were meeting DOD’s force health protection and surveillance requirements for servicemembers deploying in support of OEF and OJG, we identified DOD and each service’s overall deployment health surveillance policies. We also obtained the specific force health protection and surveillance requirements applicable to all servicemembers deploying to Central Asia in support of OEF from the U.S. Central Command and these requirements for all servicemembers deploying to Kosovo in support of OJG from the U.S. European Command. We tested the implementation of these requirements at selected Army and Air Force installations. To identify locations within each service where we would test implementation of the policies, the Assistant Secretary of Defense for Health Affairs requested the services to identify, by military installation, the number of active duty servicemembers who met the following criteria: For OEF, those servicemembers who deployed to Central Asia for 30 or more continuous days to areas without permanent U.S. military treatment facilities following September 11, 2001, and redeployed back to their home unit by May 31, 2002. For OJG, those servicemembers who deployed to Kosovo for 30 or more continuous days to areas without permanent U.S. military treatment facilities from January 1, 2001, and redeployed back to their home unit by May 31, 2002. Based on deployment data obtained from the services, we decided to limit our testing of the force health protection and surveillance policy implementation to selected Army and Air Force military installations with the largest numbers of servicemembers meeting our selection criteria (described above). We limited our review of medical records for servicemembers deploying in support of OJG to the two Army locations. We decided not to review Navy installations because there were only small numbers of servicemembers who met our selection criteria. We decided not to review Marine Corps installations because officials at the Marine Corps headquarters had difficulty identifying the number of servicemembers who went ashore 30 or more continuous days consistent with our selection criteria. The largest deployers for OEF and OJG were selected and are listed below: 10th Mountain Division, Fort Drum, N.Y. 101st Airborne Division, Fort Campbell, Ky. Travis Air Force Base, Calif. Hurlburt Field, Fla. 10th Mountain Division, Fort Drum, N.Y. 101st Airborne Division, Fort Campbell, Ky. For our medical records review, we selected statistical samples of servicemembers at the selected installations to be representative of those deploying from those military installations for those specific operations. For various reasons, medical records were not always available for review. We, therefore, sampled without replacement, to choose additional records when we were unable to meet our sampling threshold of cases for review. Specifically, there were five reasons identified for not being able to physically secure the servicemember’s medical record for review: 1. Charged to patient. When a patient visits a clinic (on-post or off-post), the medical record is physically given to the patient. The procedure is that the medical record will be returned by the patient following their clinic visit. 2. Expired term of service. Servicemember separates from the military and their medical record is sent to St. Louis, Missouri, and therefore not available for review. 3. Record is not accounted for by the medical records department. 4. Permanent change of station. Servicemember is still in the military, but has transferred to another base. Medical record transfers with the servicemember. 5. Temporary duty off site. Servicemember has left military installation, but is expected to return. The temporary duty is long enough to warrant that the medical record accompany the servicemember. The sample size for deployments was determined to provide 95 percent confidence with a 5-percent precision. The number of servicemembers in our samples and the applicable universe of servicemembers for the OEF and OJG deployments at the installations visited are shown in table 2. At Fort Campbell, there were only 333 servicemembers identified as having met our criteria based on a redeployment date of May 31, 2002; however, only 8 charts were available for review due to rotation of soldiers to other military locations or departure from the military. It was, therefore, necessary to extend our redeployment date to October 31, 2002. Doing so provided an additional 2,953 servicemembers who met all criteria except for a redeployment by May 31, 2002. At Fort Campbell, there were 92 servicemembers who deployed in support of OJG and met our selection criteria if we extended the redeployment date to October 31, 2002. Because the number of servicemembers for OJG at Fort Campbell was small, we reviewed the medical records for all of servicemembers who were still at Fort Campbell. At each sampled location, we examined servicemember medical records for evidence of the following force health protection and deployment health-related documentation required by DOD’s force health protection and deployment health surveillance policies: Pre- and post-deployment health assessments, Tuberculosis screening test (within 1 year of deployment for OEF and 2 years for OJG) influenza (within 1 year of deployment); measles, mumps, and rubella; meningococcal (within 5 years of deployment); polio; tetanus-diphtheria (within 10 years of deployment); typhoid (within 5 years of deployment); and yellow fever (within 10 years of deployment), not required for OJG. To provide assurances that our review of the selected medical records was accurate, we requested the installations’ medical personnel to reexamine those medical records that were missing required health assessments or immunizations and adjusted our results where documentation was subsequently identified. We also requested that installation medical personnel check all possible sources for missing pre- and post-deployment health assessments and immunizations. These sources included the Army’s Soldier Readiness Check folders and automated immunization sources, including the Army’s Medical Protection System (MEDPROS) and the Air Force’s Comprehensive Immunization Tracking Application (CITA). We checked all known possible sources for the existence of deployment health assessments related to servicemembers in our samples. In those instances where we did not find a deployment health assessment, we concluded that the assessments were not completed. Furthermore, installation officials were unable to logistically access the servicemembers’ individual records of immunizations, commonly referred to as yellow-shot records that may have provided documentation for missing immunizations. Consequently, our analyses of the immunization records was based on our examination of the servicemember’s permanent medical record and immunizations that were in the Army’s MEDPROS and the Air Force’s CITA. In analyzing our review results at each location, we considered documentation from all identified sources (e.g., servicemember’s medical record, soldier readiness check folder, Army Medical Surveillance Activity, and immunization tracking systems) in presenting data on compliance with deployment health surveillance policies. To identify whether required blood serum specimens were in storage at the Armed Services Serum Repository, we requested that the Army Medical Surveillance Activity staff query the Repository to identify whether the servicemembers in our samples had a blood serum sample in the repository and the date of the specimen. To determine whether the Army and Air Force are documenting in-theater medical interventions in servicemembers’ medical records, we requested, at each installation visited for medical records review, the patient sign-in logs for in-theater medical care providers, namely the Army’s battalion aid station and the Air Force’s expeditionary medical support, when they were deployed to central Asia in support of OEF and for the two Army installations we visited that deployed in support of OJG. Officials were unable to locate or access the logs at all of our selected installations except for Fort Drum for the OEF deployment. Consequently, we were able to perform our planned examination for this objective at only Fort Drum for the OEF deployment. From these logs, we selected a random sample of 36 patient visits from one battalion aid station and 18 patient visits from another battalion aid station. We did not attempt to judge the importance of the patient visit in making our selections. For the selected patient visits, we then reviewed the servicemember’s medical record for any documentation—such as the Army’s Standard Form 600—of the servicemember’s visit to the battalion aid station. To determine whether the Army and Air Force’s deployment health-related records are retained and maintained in a centralized location, we requested that officials at the Army Medical Surveillance Activity (AMSA) query the AMSA database for the servicemembers included in our samples at the selected Army and Air Force installations. For servicemembers in our samples, AMSA officials provided us with copies of deployment health assessments and immunization data found in the AMSA database. We analyzed the completeness of the AMSA database by comparing the deployment health assessments and the pre-deployment immunization data we found during our medical records review with those in the AMSA database. Since Air Force special operations force units use the Hurlburt Field, we also requested the U.S. Special Operations Command (SOCOM) to query their Special Operation Forces Deployment Health Surveillance System database for servicemembers in our sample at Hurlburt Field for deployment health assessments and pre-deployment immunization data. We then compared the data identified from the SOCOM and AMSA queries with the data we found during our medical records review. To determine whether DOD has corrected problems related to the accuracy and completeness of databases reflecting which servicemembers deployed to certain locations, we interviewed officials within the Deployment Health Support Directorate and the Defense Manpower Data Center and reviewed documentation related to the completeness of deployment databases and planned improvements in capabilities. Our review was performed from June 2002 through July 2003 in accordance with generally accepted government auditing standards. Appendix II: Comments from the Department of Defense Appendix III: GAO Contact and Staff Acknowledgments GAO Contact Acknowledgments In addition to the individual named above, Steve Fox, Rebecca Beale, Lynn Johnson, William Mathers, Terry Richardson, Kristine Braaten, Grant Mallie, Herbert Dunn, and R.K. Wild made key contributions to this report.

Summary: Following the 1990-91 Persian Gulf War, many servicemembers experienced health problems that they attributed to their military service in the Persian Gulf. However, a lack of servicemember health and deployment data hampered subsequent investigations into the nature and causes of these illnesses. Public Law 105-85, enacted in November 1997, required the Department of Defense (DOD) to establish a system to assess the medical condition of service members before and after deployments. GAO was asked to determine whether (1) the military services met DOD's force health protection and surveillance requirements for servicemembers deploying in support of Operation Enduring Freedom (OEF) in Central Asia and Operation Joint Guardian (OJG) in Kosovo and (2) DOD has corrected problems related to the accuracy and completeness of databases reflecting which servicemembers were deployed to certain locations. The Army and Air Force--the focus of GAO's review--did not comply with DOD's force health protection and surveillance policies for many active duty servicemembers, including the policies that they be assessed before and after deploying overseas, that they receive certain immunizations, and that health-related documentation be maintained in a centralized location. GAO's review of 1,071 servicemembers' medical records from a universe of 8,742 at selected Army and Air Force installations participating in overseas operations disclosed that 38 to 98 percent of servicemembers were missing one or both of their health assessments and 14 to 46 percent were missing at least one of the required immunizations. DOD also did not maintain a complete, centralized database of servicemembers' medical assessments and immunizations. Health-related documentation missing from the centralized database ranged from 0 to 63 percent for pre-deployment assessments, 11 to 75 percent for post-deployment assessments, and 8 to 93 percent for immunizations. There is no effective quality assurance program at the Office of the Assistant Secretary of Defense for Health Affairs or at the Army or Air Force that helps ensure compliance with policies. GAO believes that the lack of such a program was a major cause of the high rate of noncompliance. Continued noncompliance with these policies may result in servicemembers deploying with health problems or delays in obtaining care when they return. Finally, DOD's centralized deployment database is still missing the information needed to track servicemembers' movements in the theater of operations. By July 2003, the department's data center had begun receiving location-specific deployment information from the services and is currently reviewing its accuracy and completeness.

gov_report

en

Summarize: Background VHA, an administration of VA, is primarily a direct service provider of primary care, specialized care, and related medical and social support services to veterans through an integrated health care system. Headed by the Under Secretary for Health, VHA employed approximately 180,000 health care professionals to serve about 4.3 million veterans in fiscal year 2002. VHA’s fiscal year 2002 budget included $21.3 billion in discretionary funds from the VA/HUD (Department of Housing and Urban Development) appropriations act and an additional $142 million from an emergency supplemental enacted in August 2002. VHA developed its six strategic goals to support VA’s GPRA goals. These strategic goals are as follows: put quality first until first in quality; provide easy access to medical knowledge, expertise, and care; enhance, preserve, and restore patient function; exceed patients’ expectations; maximize resource use to benefit veterans; and build healthy communities. VHA’s strategic planning document describes strategies to show how each goal will be met. The administration then develops performance measures to support the strategies identified. VHA is headquartered in Washington, D.C. and has 21 Veteran Integrated Service Networks (networks) located throughout the country. The networks are the basic budgetary and decision-making units of VA’s health care system. They have responsibility for making a wide range of decisions about health care delivery options, including contracting with private providers for health care services and generating revenue by selling excess services. A network director, who reports to the Deputy Under Secretary for Health for Operations and Management, heads each network. This organization is illustrated in figure 1, with offices we talked to regarding VHA’s budget and planning processes shaded. The VHA Office of Quality and Performance develops and recommends performance measures (mentioned above) to the Under Secretary for Health. A Performance Management Work Group, comprised of a variety of VHA staff with different subject matter expertise, provides overall guidance with regard to the measures and helps to prioritize them. VHA’s Office of Policy and Planning prepares VHA’s contribution to VA’s 5-year Strategic Plan, as well as the Network Strategic Planning Guidance, which is used by networks to prepare their strategic planning documents. Among other responsibilities, VHA’s Office of Finance is responsible for policy and operational issues relating to budget formulation and execution, financial management, and financial analyses. As a consequence of VHA’s field structure reorganization, decision making is currently more decentralized. In 1995, the 172 independent VA Medical Centers were reorganized into 22 networks, headed by network directors. Network directors are accountable for a variety of functions, such as contracting, budgeting, and planning for the medical facilities within their purview. Under this reorganization, VHA management anticipated that network directors could manage the distribution of the networks’ resources to maximize the advantages to veterans within their service areas. Furthermore, the administration expected to perform less operational decision making and oversight at the central office level. Along with the decentralization, VHA shifted its service delivery focus from inpatient hospital care to outpatient care; between fiscal years 1995 and 2001, the average number of hospital inpatients declined from 31,137 per day to 13,452 per day. The number of annual outpatient visits increased from 26 million to 41 million during the same period. Scope and Methodology To address the report’s objectives, we interviewed senior officials in VHA and VA to find out how they used performance information in the budget process. We reviewed several network managers’ performance contracts and information on network performance measures to learn about the level of accountability VHA expects from its networks. We reviewed VA guidance on preparing budget requests, budget submissions, and other related documents for information on the budget process and the use of performance information. To learn about the planning process within VHA and assess its integration with the budget process, we read VA strategic plans and other planning documents. We attended congressional hearings and reviewed related documents to learn about VHA’s budget requests, use of performance information, and VA/Department of Defense resource sharing. We did not assess the appropriateness of VHA’s performance measures or budget requests, or the accuracy of VHA’s performance management information. We conducted interviews in Washington, D.C. with senior officials from the VA Office of Budget, VHA Office of Finance, VHA Office of Policy and Planning, VHA Office of Quality and Performance, VHA Management Support Office, and the Liaison Staff Office to learn about VHA’s budget process and performance measures. Because we found the most evidence of a linkage between budget and performance during budget execution at the network level, we focused our work on that process at that level. We selected two networks—Network 2, in Albany, New York and Network 13, in Minneapolis, Minnesota—that VHA officials believed made the best use of performance information in managerial decision making. Networks 13 and 14 were combined and renamed Network 23 in January 2002, leaving 21 operational networks; there is a break in numerical sequence. We chose to focus on Network 13 rather than Network 23 since the structure of Network 23 had not yet been finalized at the time of our review, and we were interested in looking at processes that were already in place. Just as findings at individual federal agencies cannot be generalized across all agencies, the 2 networks selected for review are not representative of the other 19 networks. However, the observations of the network managers we interviewed are useful in understanding the different approaches taken to integrate budget and performance. We reviewed network-specific budget and planning documents, such as strategic plans, annual performance plans, performance reports, and tactical plans for the two networks we visited. We interviewed over 20 network officials, including senior network management, care and patient service line managers, facility managers, information technology managers, quality management officials, and strategic planners to learn about network structures and the use of performance information in decision making at the networks. We also reviewed a number of background documents on administration initiatives and performance budgeting implementation, as well as recent public administration literature and GAO reports for general background and context. We conducted our work between January 2002 and June 2002 in accordance with generally accepted government auditing standards. Budget Formulation and Planning Efforts Are Centrally Managed, While Budget Execution and Planning Are Linked in Networks Although VHA’s budget formulation and planning processes are both centrally managed, they are not closely linked. The agencywide budget request is based primarily on the previous year’s appropriations with some adjustments for workload and new policies. Distribution of funds to networks is largely driven by a system that is heavily based on the veteran population served at each network. Budget and performance are more clearly linked at the networks we visited. See figure 2 for an overview of VHA’s budget and planning processes. VHA’s Budget Formulation and Planning Processes Are Centralized but Not Integrated VHA reported that its budget formulation processes for fiscal years 2002 and 2003 were developed centrally with limited input from the networks and reflected an incremental approach, with some adjustments in fiscal year 2003 for projected workload increases and administrative efficiency assumptions. Prior to the development of the fiscal year 2003 budget, senior budget officials told us that VHA sought selected information from the networks, such as estimates of collections and long-term care expenditures. In preparing the budget that would eventually be included in VA’s submission to OMB, VHA generally used the current appropriations levels as a baseline and added an adjustment for workload, as well as an increase for new programmatic initiatives. One VHA official commented that the process was very “top down.” For fiscal year 2003, the main change to this process was the introduction of some enrollment growth projections from an actuarial model and administrative efficiency adjustments for reducing resource requirements. VHA receives guidance on how to formulate its budget through a budget call memorandum issued by VA in April. This memorandum includes VA strategic goals and objectives and stresses the need to focus on outcome and performance goals and measures. Once VHA’s Office of Finance formulates the administration’s budget, it is sent to VA’s Office of Budget where the VHA request is reviewed and recommendations are made by VA senior staff, culminating in a department budget request for VHA and the other two administrations. This submission is sent to OMB in September. Decisions made by OMB are incorporated into the President’s Budget presented to the Congress. Following congressional action and enactment of the appropriations bill, OMB apportions and VA allots the funding provided in the VHA appropriations, thus beginning the execution phase of VHA’s budget cycle. While the VHA-related information in VA’s annual Performance Plan describes goals, strategies, and performance measures, the relationship to the budget formulation process is unclear. VHA officials told us that they use strategic planning information as source material for departmental reports (e.g., the Accountability Report and VA’s Annual Performance Plan), to review networks’ policies for consistency, and generally to have the information on hand in an organized format. VA’s Annual Performance Plan outlines resource requirements by strategic goal, and each strategic goal is accompanied by performance goals and measures. However, a VHA official told us that planning documents are typically finalized after VHA’s budget is formulated. VHA Resource Allocation Is Largely Formula-Driven About 90 percent of VHA’s medical care appropriation, which is approximately 86 percent of VHA’s total budgetary resources, is allocated to networks through the Veterans Equitable Resource Allocation system (VERA), which uses a formula that calculates resource allocation based on workload. The remainder of the appropriated funds is allocated to networks either through Specific Purpose Funding, which is designated for certain programs such as state home funds or Vietnam veterans’ readjustment counseling. Monies in the no-year Medical Care Collections Fund (MCCF), as well as other small nonappropriated funds, are also available to the networks. Network Planning and Budget Execution Processes Are More Closely Related Decisions regarding resource allocation and planning are closely aligned at both networks we visited. The same officials are involved in strategic planning and budget execution, and network-produced documents show some alignment between planning efforts and resource allocation. The Office of Policy and Planning prepares VHA Network Strategic Planning Guidance, directing networks on how to develop their individual strategic plans. According to the guidance, strategic plans must associate performance measures with each strategic objective. For example, the fiscal years 2003-2007 Guidance for Strategic Objective 1, “Put Quality First Until First in Quality,” identifies the first strategy as, “Systematically measure and communicate the outcomes and quality of care,” and the related performance measure as “Improve performance on the Chronic Disease Care Index II.” Networks must then identify their plans to meet each performance measure. VHA’s Office of Finance issued guidance that required networks to provide financial or operating plans for a range of possibilities. According to officials, networks prepare plans in anticipation of final appropriations actions. Once VHA receives its appropriations and VERA allocations are calculated, networks submit plans to the Office of Finance that lay out the networks’ spending plans for their VERA funds. The two networks we visited, Network 2 and Network 13, are structured somewhat differently with regard to resource allocation authority. At Network 2, service delivery is organizationally divided into Care Lines; Care Line Directors have resource allocation authority across all medical facilities in the network. For example, according to network officials, the Geriatrics and Extended Care manager can make resource allocation decisions concerning nursing home care at all Network 2 facilities. Network 13, on the other hand, is structured around Patient Service Lines (PSLs). PSL Chief Executive Officers (CEOs) share resource allocation authority with the Chief Operating Officers (COO) at each medical center. PSL CEOs make allocation decisions for the facilities that support their PSL at the beginning of the fiscal year, while day-to-day smaller resource decisions during the fiscal year are handled primarily by each medical center COO. CEOs and COOs collaborate on larger budget-related decisions across the PSL. The subject of the resource decision determines which PSL CEO is involved; for example, the PSL CEO for Mental Health is involved with decisions regarding psychiatric care. Annual budgets are developed by the PSL CEOs in conjunction with site COOs and Chief Financial Officers (CFOs). At Network 2, network leadership works with Care Line Directors in developing and prioritizing strategic goals and targets. Network 2’s strategic plan shows a link between VHA strategies and performance measures, and network-specific actions to achieve them. (See fig. 3 for an example of this linkage.) The plan also shows how expected increases in annual funding will be used by program line. Network 13 senior managers told us about annual 2-day tactical planning meetings that were designed to provide an outlet for stakeholders to plan and share information on performance and strategic planning, cost information, performance measures, successful practices, and lessons learned. Participants include PSL CEOs, a PSL COO, PSL managers under COO control, union representatives, and congressional stakeholders. A network official stated the purpose of including managers with resource allocation authority in tactical planning meetings was to strengthen the link between the processes of resource allocation decisions and planning. Performance Information Influences Resource Allocation Decisions in a Variety of Ways at These Networks Integrating performance information into resource allocation decisions is apparent at the network level during budget execution. At the two networks we visited, managers told us that they use an internal data system that compares cost and performance data across facilities as a tool to make resource allocation decisions. Network performance is monitored by VHA, and networks establish their own processes to monitor their performance. Network managers also use various communication methods, both within their networks and across other networks, to share information on performance measures and ways to meet those measures. Managers reported that they were accountable for performance and provided examples where they used performance information to make resource allocation decisions. Cost and Performance Data Used in Managerial Decision Making Network managers told us that they use data from the Decision Support System (DSS) to make resource allocation decisions to their facilities and programs. DSS is an executive information system designed to provide VHA managers and clinicians with data on patterns of patient care and patient health outcomes. It is also used to analyze resource utilization and the cost of providing health care services. For example, a manager in Network 2 said that he uses DSS data for comparisons of facilities, population and market share data, and veterans’ length of stay in inpatient units. Since veterans are staying in inpatient units for fewer days in certain facilities, the manager has been able to reallocate money across facilities because of DSS data. Networks’ Performance Monitored As we noted in “Results-oriented Budget Practices in Federal Agencies,” it is important for agency management to monitor performance. VHA leadership uses several methods to monitor network performance and hold network officials accountable for that performance. At its quarterly meeting, the VHA Executive Leadership Council (ELC), which includes the deputy secretary, managers from all three administrations, network directors, other key staff, interest groups, and the public, monitors the status of performance measures at each network. In addition, the Deputy Secretary of Veterans Affairs began holding monthly meetings in late 2001 with the Under Secretary of VHA and all the network directors. At these meetings, the senior officials provide information on each network’s successes in meeting VHA-established performance measures and share best practices in meeting performance measures. Networks must provide remedial action plans at these meetings for measures that are not being met. For example, one network was deemed deficient in testing patients for Hepatitis C. Its action plan included a review of patients who had not been tested and an electronic clinical reminder to help service providers identify patients who have risk factors but were not tested. To make sure network directors understand the importance VHA places on performance, directors sign an annual performance agreement with the Under Secretary for Health called the Network Performance Plan. The agreement includes expectations regarding VHA-level performance measures and their associated strategic goals. According to VHA guidance, a network director’s appraisal is affected by his or her network’s performance in relation to agency goals. As a result, the director’s compensation may also be affected. For example, in 2001 a network director received a bonus because his network exceeded VHA-established performance goals. Networks Establish Their Own Processes to Monitor Performance The two networks we visited each had its own ELC to review performance measures on a network level and commissioned task teams to work on areas where performance has not met the intended goal. Quality Management Officers (QMOs) also serve as performance monitors. The QMO at Network 2 keeps track of the network’s action plans to improve upon deficient performance measures, and reports on performance-related data at the Transforming Systems Performance & Quality Council (TSPQ), a forum to address issues across care lines and facilities to work collaboratively toward addressing performance measure issues, quality management, information systems, and related operational issues. Membership includes top network management, care line managers, and network office staff. Network 2 uses its Web site in various ways to maintain up-to-date information on performance measures. For example, Network 2 managers told us about the Web-based Pulse Points, which are performance indicators that assist management in monitoring achievement of performance measures. Additional performance-related information is available to network staff on the intranet. Communication Important to Help Managers Meet Performance Measures Sharing information about lessons learned and strategies to achieve performance measures can lead to more informed resource allocation decisions. Between networks, managers have a number of opportunities to learn from each other via regularly scheduled meetings and conference calls. Network managers told us about periodic meetings where they interact with managers from other networks and share lessons learned with regard to cost-saving measures and approaches to performance measures. Within a network, staff may also use a variety of communication tools to share information to improve performance. For example, two Network 2 care line managers jointly established a team to discuss ways that the network could better meet performance goals for length-of-stay rates. This team, which spanned multiple care lines, worked on the issue and communicated its recommendations quarterly. Also, VA sponsors a “Lessons/Innovations” database on the Internet, where network managers can read ideas for performance improvement. Network Managers Reallocate Resources in Response to Performance Measures In both of the networks we visited, managers provided examples where performance information and the responsibility to meet performance targets affected the way in which they allocated resources. The examples incorporated a number of different approaches to improve performance, including investing in telemedicine and technology advancements, resource reallocation, low-tech methods to improve performance, and the use of outside contractors. Performance target: 100 percent of diabetic veterans should receive retinal eye exams to decrease the potential incidence of blindness. Network manager approach: Use telemedicine and special equipment to allow diabetic veterans who receive care at locations that do not have ophthalmologists the ability to have their exam results read by qualified specialists. An initial investment of network resources in advanced telemedicine techniques led to an increased percentage of diabetic veterans receiving a necessary test. To reduce the potential for blindness later in life, all diabetic veterans are supposed to have retinal eye exams to monitor their vision. However, this requires the services of an ophthalmologist who must interpret the exam results. The network did not have resources to maintain an ophthalmologist on staff at each site, so many diabetics were not being tested. A Network 2 manager found that a particular piece of equipment could record test results, then transmit them to an ophthalmologist at another location. Thus, the network invested resources in a number of these machines for Community Based Outpatient Clinics (CBOCs) to use, thereby increasing the network’s capacity for meeting this performance target. Performance target: Annual cost per patient must be below a given threshold. Network manager approach: Moved $100,000 from one facility that was not taking on as many patients as expected to another facility with an increased workload. A manager at Network 13 noted that facilities are expected to keep their average cost per patient down. Regular monitoring revealed that one facility was not taking on as many patients as planned, which led to higher average costs. To reinforce his expectation that this performance target should be met, the manager chose to transfer financial resources from this facility to another facility that required additional staffing to meet other performance targets. According to the manager, the facility that received the funds was able to improve its outcomes in the targeted area. Performance measures: Reducing the number of falls out of bed and the use of restraints. Network manager approach: Buying lower beds (9” off ground). A Network 13 manager noted that veterans were falling out of their beds and thus incurring injuries, and the manager searched for a way to reduce this incidence. He also wanted to reduce the use of restraints, another performance measure. Based on staff recommendations, the manager agreed to invest in beds that were only 9 inches off the ground. This investment prevented more serious injuries from occurring, reduced the need for restraints, and directly improved the network’s performance in these areas. Performance target: Veterans should be able to obtain appointments with mental health professionals within 30 days of request. Network manager approach: Reviewed various staff practices and made recommended improvements. A PSL manager in Network 13 noted that wait times for veterans to obtain appointments with mental health specialists exceeded the performance target. The manager hired an outside consultant who looked at a variety of factors, including (1) how physicians’ time was being spent, (2) physicians’ practices regarding rescheduling appointments, and (3) hiring psychiatrists instead of contracting for them. According to the manager, after the network adopted the consultant’s recommendations, including hiring (instead of contracting for) psychiatrists and hiring administrative staff to prescreen patients, Network 13 met the performance target by eliminating wait time completely. Challenges VHA has undergone a cultural shift over the past 7 years that has helped to integrate budget and performance, but managers face continuing challenges to further integration and in defining areas for improvement. The agency’s budgeting and planning processes are not directly linked, so opportunities are missed to fully use planning information in the budget process. Additionally, VHA does not use the most complete information available when making resource allocation decisions. Planning and Budgeting Linkage Could Improve VHA’s planning and budgeting processes are not fully integrated (see fig. 2 for an overview of these processes). VHA officials acknowledged the offices in charge of these processes did not work closely together in the past, but steps are being taken to improve this linkage. For example, a member of VHA’s Office of Finance now works in the Office of Policy and Planning on the agency’s demand model, which will be used to project costs for fiscal year 2004. According to VHA, this model projects workload for VHA nationwide and was partially used to prepare the fiscal year 2003 budget request. Future workload is projected through the use of a detailed formula that includes enrollment, anticipated utilization, and reliance on VA services. It does not assume an incremental increase over current workload. Performance Information is Available but Not Included in Resource Allocation Decisions VHA does not include the most complete information available when allocating resources. As we noted in VA Health Care: Allocation Changes Would Better Align Resources with Workload (GAO-02-338), VA does not adequately account for important variations in patients’ health care needs across networks nor does it include all veterans who use health care services in its resource allocation decisions. Without complete information, it is difficult for agencies to consider fully the relative priorities of programs and activities and, when funding tradeoffs are necessary, where adjustments can be best made. Producing reliable funding estimates requires an agency to include reasonable assumptions about factors affecting program costs or budgetary resources, assess the accuracy of previous estimates, and if necessary, make appropriate adjustments to its estimating methods. Agency Comments and Our Evaluation In its comments, VA agreed with our observations but asserted that our report does not give the reader an adequate grasp of the depth and breadth of managing such a large health care system. VA also included three enclosures: the first was intended to clarify certain points in the draft report, the second provided information on VA’s actuarial model, and the third outlined VA’s new budget account structure. In the first enclosure, VA suggested three clarifications regarding our report language. 1. VA stated that the fiscal year 2003 budget was based on actuarial projections of workload broken down by specific disease and veteran priority level using prior years’ costs; it also noted that administrative efficiency assumptions were further included for reducing resource requirements. During our interviews, officials told us that the process was generally incremental, but actuarial projections were used only to calculate potential increases in workload for fiscal year 2003; these projections were not used to reassess the base. For the fiscal year 2004 budget and beyond, officials expected to use the actuarial projections to calculate the entire workload, not just the potential increase. We made changes in our report language to reflect this process. 2. VA noted that it does not include all Priority 7 veterans in its VERA calculations because it does not want to provide financial incentives that encourage network managers to provide care to Priority 7 veterans at the expense of higher-priority veterans. As we recommended in our February 2002 report, networks with a disproportionately large number of Priority 7 veterans already have fewer resources under VERA to treat higher-priority veterans on a per-patient basis. To remedy this problem, we recommended that VA align measures of workload with actual workload served, regardless of veteran priority group. Doing so will help provide comparable resources for comparable workload. Thus, we maintain that complete information allows agencies to better consider the relative priorities of programs and activities. 3. VA also noted that the funds it received under the Emergency Supplemental appropriation were not intended for homeland security- related activities. We changed our report language appropriately. VA’s second enclosure was a report that describes the actuarial model it uses to project the demand for health services. This report was prepared for the Senate Appropriations Committee, in response to a congressional mandate identified in S. Rpt. 107-156. During our fieldwork, we were told that there was no documentation available regarding this model. We received the documentation when the draft report was sent for agency comment and therefore did not review the model and its ability to project VA’s workload. The third enclosure focuses on VA’s budget account restructuring for its fiscal year 2004 budget submission. VA notes that this structure will allow it to more readily determine the full cost of each of VA’s programs and make resource decisions based on programs and their results rather than on other factors. We did not review this new structure or its ability to more effectively link resources with results since the outcome will not be available until the administration’s budget proposal is released in early 2003. As agreed with your office, we will distribute copies of this report to the Secretary of Veterans Affairs, appropriate congressional committees, and other interested parties. We will make copies available to others upon request. In addition, the report will be available at no charge on the GAO Web site at http://www.gao.gov. Please contact me on (202) 512-9573 or Denise Fantone, Assistant Director, on (202) 512-4997 if you or your staff have any questions about this report. Major contributors to this report are Kimberly Gianopoulos, Kelli Ann Walther, and James Whitcomb. Comments From the Department of Veterans Affairs GAO’s Mission The General Accounting Office, the investigative arm of Congress, exists to support Congress in meeting its constitutional responsibilities and to help improve the performance and accountability of the federal government for the American people. GAO examines the use of public funds; evaluates federal programs and policies; and provides analyses, recommendations, and other assistance to help Congress make informed oversight, policy, and funding decisions. GAO’s commitment to good government is reflected in its core values of accountability, integrity, and reliability. Obtaining Copies of GAO Reports and Testimony The fastest and easiest way to obtain copies of GAO documents at no cost is through the Internet. GAO’s Web site (www.gao.gov) contains abstracts and full- text files of current reports and testimony and an expanding archive of older products. The Web site features a search engine to help you locate documents using key words and phrases. You can print these documents in their entirety, including charts and other graphics. Each day, GAO issues a list of newly released reports, testimony, and correspondence. GAO posts this list, known as “Today’s Reports,” on its Web site daily. The list contains links to the full-text document files. To have GAO e-mail this list to you every afternoon, go to www.gao.gov and select “Subscribe to GAO Mailing Lists” under “Order GAO Products” heading. Order by Mail or Phone To Report Fraud, Waste, and Abuse in Federal Programs Public Affairs

Summary: Encouraging a clearer and closer link between budgeting and planning is essential to improving federal management and instilling a greater focus on results. Through work at various levels within the organization, this report on the Veterans Health Administration (VHA)--and its two companion studies on the Administration on Children and Families (GAO-03-09) and the Nuclear Regulatory Commission (GAO-03-258)--documents (1) what managers considered successful efforts at creating linkages between planning and performance information to influence resource choices and (2) the challenges managers face in creating these linkages. VHA's budget formulation and planning processes are centrally managed, but are not closely linked. Resource distribution to VHA's health care networks is mostly formulaic, determined primarily by the distribution of the veterans being served. VHA offices involved in budget formulation and strategic planning provide guidance to health care networks in developing their financial and strategic plans. Integrating performance information into resource allocation decisions is apparent at the health care network level during budget execution. Health care network managers told us that they use an internal data system as a tool to decide how to allocate resources to their facilities and programs. They also use various communication methods to share information on performance measures, and are held responsible for meeting those measures. Network managers provided specific examples where performance information influenced their resource allocation decisions. For example, one performance target specifies that all diabetic veterans are expected to receive retinal eye exams. An ophthalmologist must interpret the results of such an exam; however, most outpatient clinics do not have the resources to maintain an ophthalmologist on staff. One network invested in machines that record test results and transmit them to an ophthalmologist at another location, thereby increasing the network's capacity for meeting this performance target. While budget and performance integration has improved, VHA managers still face additional challenges. VHA's budgeting and planning processes are not directly linked, but VHA officials noted that steps are being taken to better integrate them. Also, VHA does not use the most complete information available when making resource allocation decisions to its health care networks, so the link between resources and results could be improved.

gov_report

en

Summarize: By. Jonathan Petre And Andy Gardner For The Mail On Sunday. THE only Hurricane that fought in the Battle of Britain and is still flying today is up for sale for £2.5 million – more than 30 years after it was found as scrap in India. The aircraft, which regularly takes part in flypasts, was restored by a vintage car buff who discovered the wreck by chance while seeking old Rolls-Royces. But retired businessman Peter Vacher is now selling the 1940 Hawker Hurricane Mk 1 through an American dealer, fuelling fears that it could be lost to Britain. Scroll down for video. Sole survivor: The Hurricane R4118 (pictured) is thought to be the last plane from the Battle of Britain still flying. Hurricane R4118 flew 49 combat sorties during the worst days of the fighting and shot down five enemy aircraft. Hurricanes, alongside Spitfires, were at the centre of the country’s heroic defence of the skies in 1940 and had a crucial role until the end of the war, but the vast majority were then scrapped. Mr Vacher, 72, said: ‘This Hurricane is a one-off. There is no other British plane like it from the Battle of Britain that is flying today. It was a labour of love to recover and restore it and I have enjoyed watching it fly for the past ten years. Heroic: Wing Commander Bob Foster, who was responsible for three of the plane’s five ‘kills’ in the Battle of Britain died last month aged 94. ‘However, it is time for somebody else to take it on – and my hope is that it will stay in the UK or in Europe. Obviously, there is a possibility that it could go further afield. But it is such an iconic British plane. The sight and sound of it is the Battle of Britain. The public absolutely love it.’ The former printer spotted the aircraft’s rotting hulk in 1982 on a visit to a university at Benares, north-east India, but did not immediately realise what it was. The plane had been given to engineering students in 1947 after the war in the Far East came to an end. The fuselage was lying in a compound with the propeller, wings and tailplane spread out over the ground. Mr Vacher, of Abingdon, Oxfordshire, said: ‘It looked like a pile of junk, but once I had found out what it was, I became obsessed with restoring it.’ He returned to India in 1996 and, after a long battle with local bureaucrats, bought the fighter for £25,000 in 2001 and shipped it to Britain. It took four years and 32,000 hours of restoration work, including renovating a Rolls-Royce Merlin III engine, before the Hurricane was once again airworthy. It has since flown in public more than 100 times. Wing Commander Bob Foster, who was responsible for three of the plane’s five ‘kills’ in the Battle of Britain, died last month aged 94. A spokesman for the Battle of Britain Memorial Trust said: ‘It would be disappointing if Bob Foster’s Hurricane were to leave the country. It is very special and it would be a shame if it could no longer be seen over the UK.’ The aircraft is being offered for sale through Platinum Fighters. Final Fighter: Online ad for the Hurricane flown by Bob Foster, right, that's still in flying condition

Summary: Legendary plane was found as scrap in India more than 30 years ago. Bought by Peter Vacher, of Abingdon, Oxfordshire, for £25,000 in 2001. Retired businessman and car buff, 72, spent 32,000 hours restoring model. Says it's the only plane from the Battle of Britain that is still flying today. But now he's selling it through an American dealer - meaning it could be lost to Britain forever.

cnn_dailymail

en

Summarize: Actor Jonathan Goldsmith attends the Kentucky Derby in 2011. (Photo: Michael Loccisano, Getty Images) WASHINGTON — TV's "most interesting man in the world," known in Dos Equis beer commercials for achieving the wildly impossible, probably could eliminate landmines by himself — using only his beard. But actor Jonathan Goldsmith, who has played the cultural icon for nearly nine years, is looking for help from others. He's raising funds for landmine removal in Cambodia, where accidents involving buried explosives have spiked this year, through a contest to be his guest for a day in Vermont. He won't be in character, but he'll make it interesting. "We're going to spend a day doing some things — off-road driving and some other little events, one being falconry," said Goldsmith, who turns 76 on Friday. Goldsmith has volunteered for the Mines Advisory Group since visiting the non-profit group's operations in Vietnam last year. MAG has worked in more than 35 countries and shared the Nobel Peace Prize in 1997. The contest will benefit the organization. "It's a terribly important charity," said Goldsmith of Manchester, Vt. "Accidents are up. Funding is down. Kids are getting hurt. I want to do whatever I can to help and bring attention to it." In the popular Dos Equis ads — an Internet meme — Goldsmith's dashing, cigar-smoking character can be found dog-sledding to a black-tie party, saving a firefighter from a burning building or releasing a growling bear from a trap. A narrator claims, "He's trained canaries in the art of falconry" or "He can speak Russian … in French." Goldsmith often ends the ads relaxing in a lounge while surrounded by young women. He delivers his classic sign-off, "Stay thirsty, my friends," in a deep Spanish-accented voice inspired by his late friend, the actor and director Fernando Lamas. Through his advocacy for landmine removal, Goldsmith fills a void left by other celebrity advocates, most notably the late Princess Diana, said Jamie Hathaway, a MAG consultant. "Ironically, when much of the world has lost interest in the movement, comes the'most interesting man in the world' to help us out," said Hathaway, a friend of Goldsmith's who got him involved with MAG. In April, Goldsmith joined Sen. Patrick Leahy, D-Vt., for a reception and photo exhibit honoring the group's 25 years of work. Leahy, a decades-long proponent of landmine removal efforts, said Goldsmith has "invested his time, his talent and the intangible capital of his star power to bring attention to these vital missions." The United States is the biggest donor to mine removal efforts. This week, the Obama administration moved closer to compliance with a global treaty banning landmines, announcing the United States will not use mines outside the Korean Peninsula. The United States is among 34 United Nations members that haven't signed the treaty. Casualties from mines and other explosives totaled 3,628 in 2012, according to the Landmine and Cluster Munition Monitor, an initiative of the International Campaign to Ban Landmines. That's the lowest total since the monitor began recording casualties in 1999. Read or Share this story: http://usat.ly/1sv3g7F Starting in 1996, Alexa Internet has been donating their crawl data to the Internet Archive. Flowing in every day, these data are added to the Wayback Machine after an embargo period. WE BELIEVE IN EMBELLISHING YOUR STORIES. NOT YOUR AGE. Tell us your date of birth Lady Gaga rocked some 80's style while leaving her apartment building in New York City on June 6. Eva Longoria and Felicity Huffman had a little "Desperate Housewives" reunion at the AFI Life Achievement Award: A Tribute To Jane Fonda at the Dolby Theatre in Hollywood, California on June 5. Lindsay Lohan was seen at Chiltern Firehouse in London on June 5. Scott Disick attended the 2014 FIFA World Cup McDonald's launch party at Pillars 38 on June 5 in New York City. Lena Dunham was seen rocking a "No Comment" sweater while out and about in New York City on June 5. Hilary Duff was all smiles as she ran errands on June 5 in Beverly Hills, California. Channing Tatum has some fun at the "22 Jump Street" premiere at AMC Lincoln Square Theater on June 4 in New York City. Nicole Kidman and Keith Urban attended the 2014 CMT Music Awards at the Bridgestone Arena on June 4 in Nashville, Tennessee. Carrie Underwood attended the 2014 CMT Music Awards at the Bridgestone Arena on June 4 in Nashville, Tennessee. Kendall and Kylie Jenner were seen stepping out in New York City on June 4. Bradley Cooper got whipped into shape by military personnel while filming scenes for his upcoming movie "American Sniper" in Los Angeles, California on June 4. Sarah Jessica Parker went for a walk on June 4 in New York City. Emma Roberts and Evan Peters enjoyed a romantic stroll on the beach together while vacationing in Maui, Hawaii on June 3. Kim Kardashian stopped by Craig's in West Hollywood, California to have lunch with her mom Kris Jenner on June 3. Taylor Schilling attended the Glamour Women of the Year Awards at Berkeley Square Gardens on June 3 in London, England. Miranda Lambert stopped by ABC studios for an appearance on "Good Morning America" in New York Cit on June 3. Angelina Jolie attended the "Maleficent" photo call at The Bund on June 3 in Shanghai, China. John Mayer enjoyed dinner out at Madeo Restaurant on June 2 in West Hollywood, California. Blake Lively left her hotel with designer Michael Kors and headed to the 2014 CDFA Awards on June 2 in New York City. Gwen Stefani spotted leaving a skin care clinic while out and about in Beverly Hills, California on June 2. Katie Holmes spotted picking up Jergens Natural Glow Daily Moisturizer in New York City on June 2. Pete Wentz took a test drive of the new Mario Kart 8 for Wii U in Los Angeles, California on June 2. Halle Berry attended the Huading Film Awards on June 1 at Ricardo Montalban Theatre in Los Angeles, California. Kim Kardashian was seen at LAX on June 1 in Los Angeles, California. Shailene Woodley was seen leaving the Bowery Hotel in New York City on June 1. Marion Cotillard spotted out and about with a friend in New York City on June 1. Khloe Kardashian chatted away on her cell phone as she arrived on a red eye flight at JFK airport in New York on May 31. Naomi Watts headed to the gym for a morning workout on May 30 in Brentwood, California Jane Lynch signed copies of her book "Marlene, Marlene, Queen of Mean" at the 2014 Bookexpo America at The Jacob K. Javits Convention Center on May 29 in New York City. Taylor Schilling, Jason Biggs and Laura Prepon attended a photo call to launch season 2 of Netflix exclusive series "Orange Is The New Black" on May 29 in London. Peter Facinelli and Jaimie Alexander shared a kiss after their work out at a gym in West Hollywood, California on May 29. Mariah Carey at the 2014 Fresh Air Fund Honoring Our American Hero at Pier Sixty in New York City on May 29. Farrah Abraham spotted posing for cameras in New York City on May 29. Prince William and Kate Middleton visited Strathearn Community Campus in Crieff, Scotland on May 29. "The Bachelorette" Season 1 couple Trista and Ryan Sutter stopped to pose for the cameras while out and about in New York City on May 29. Angelina Jolie and Brad Pitt at the Los Angeles premiere of "Maleficent" at the El Capitan Theatre in Hollywood, California on May 28. Chris Martin and his band Coldplay perform live at the Casino de Paris on May 28 in Paris. Hilary Duff stopped by a nail salon in Los Angeles, California on May 28. Scott Disick spotted out and about with a friend in New York City on May 28. Emily Blunt attended the premiere of "Edge Of Tomorrow" on May 28 in London. Nina Dobrev wowed at the World Music Awards 2014 at Sporting Monte-Carlo on May 27. Kristen Stewart departed on a flight at LAX airport in Los Angeles, California with her bodyguard on May 27. Amanda Seyfried, Charlize Theron and Seth MacFarlane attended a photo call to promote "A Million Ways To Die In The West" on May 27 in London, England. Jennifer Garner met a friend for lunch at Larchmont Bungalow in Los Angeles, California on May 27. Tom Cruise attended "Edge Of Tomorrow" photocall at Terrazza Civita on May 27 in Rome, Italy. Kaley Cuoco and her husband Ryan Sweeting attended Joel Silver's Annual Memorial Day Party at his home in Malibu, California on May 26. Kris Jenner and baby North West left Florence Airport after Kim Kardashian And Kanye West's wedding on May 25 in Florence, Italy. Emma Watson, in a cap and gown, attending Brown University’s 2014 Graduation Ceremony in Providence, Rhode Island on May 25. Eva Longoria had a busy morning in Los Angeles, California on May 25. Iggy Azalea arrived at The Bank nightclub at the Bellagio on May 24 in Las Vegas, Nevada. Supermodel Hilary Rhoda and her fiance former "DWTS" contestant Sean Avery celebrated Memorial Day with Tequila Don Julio cocktails from the Airstream speakeasy in the Hampton's on May 24. Hilary Duff dined out at Craig's restaurant on May 24 in West Hollywood, California. Kim Kardashian and Kanye West got married on May 24 in Florence, Italy. Uma Thurman attended the "Clouds Of Sils Maria" premiere at the 67th Annual Cannes Film Festival on May 23 in Cannes, France. Kristen Stewart, Juliette Binoche and Chloe Grace Moretz attended the "Clouds Of Sils Maria" premiere during the 67th Annual Cannes Film Festival on May 23. Kanye West, Kim Kardashian and Kris Jenner were spotted leaving an apartment on May 23 in Paris, France. Chris Martin arrived at LAX Airport to catch a flight out of town on May 22 in Los Angeles, California. Lea Michele signed copies of her book "Brunette Ambition" at Barnes & Noble Book Store at the Grove in Los Angeles, California on May 22. Jude Law at the Peace One Day Monaco Gala in support of Peace One Day's Education work in the Democratic Republic of Congo and Great Lakes region in Monaco on May 22. Pharrell Williams visited the ITV studios on May 22 in London, England. Drew Barrymore and Will Kopelman arrived at the Los Angeles premiere of "Blended" at TCL Chinese Theatre on May 21 in Hollywood, California. Jennifer Lopez wowed at the "American Idol" finale at the Nokia Theatre LA Live in Los Angeles, California on May 21. Emma Stone and Andrew Garfield went out for a coffee in New York City on May 21. Kim Kardashian and her family dined out at Costes restaurant on May 21 in Paris, France. Rosie Huntington-Whiteley attended the "The Search" premiere at the 67th Annual Cannes Film Festival on May 21. Heidi Klum and her boyfriend Vito Schnabel arrived in Nice, France for the 67th Annual Cannes Film Festival on May 21. Ali Larter shopped at Marshalls in Los Angeles, California on May 21. Tom Cruise was spotted on a night out at the Chiltern Firehouse in London, UK on May 20. Cheryl Burke at the 'Dancing With The Stars" Season 18 Official Wrap Party at the Sofitel Hotel in Los Angeles, California on May 20. Jessica Alba was all smiles while stopping by her office in Santa Monica, California on May 20. Christina Hendricks and Ryan Gosling hugged it out at the "Lost River" photocall at the 67th Annual Cannes Film Festival on May 20. Naomi Watts started her morning off with a workout at the gym on May 20 in Brentwood, California. Katie Holmes glowedin a floral Desigual dress at an NYC studio on May 20. Kate Hudson wore short shorts while out and about in New York City on May 19. Drew Barrymore and Adam Sandler cozied up at the premiere of "Blended" at the CineStar Cinema in Berlin, Germany on May 19. Kim Kardashian and her fiance Kanye West spent the day shopping at luxury brands Balenciaga, Versace and Colette on May 19 in Paris, France. Jessica Chastain attended the "Foxcatcher" premiere during the 67th Annual Cannes Film Festival on May 19 in Cannes, France. Andi Dorfman, the new Bachelorette, dropped by ABC Studios for an appearance on "Good Morning America" on May 19. Jennifer Lopez in the 2014 Billboard Music Awards Press Room held at the MGM Grand Garden in Las Vegas, Nevada on May 18. Jennifer Lawrence attended Lionsgate's "The Hunger Games: Mockingjay Part 1' party at a private villa on May 17, 2014 in Cannes, France. Matthew McConaughey and his family participated in Drew Brees', The Brees Dream Foundation charity fundraiser mimicking the Amazing Race in New Orleans, Louisiana on May 17. Naomi Watts spotted at the beach wearing Skechers GOwalk 2, while taking a break from the Cannes Film Festival on May 17. Kim Kardashian landed on a flight at LAX Airport on May 15 in Los Angeles, California. Charlize Theron and Sean Penn attended the "A Millions Ways To Die In The West" premiere held at the The Regency Village Theater in Westwood, California on May 15. Jennifer Lopez arrived on the set of "American Idol" in Hollywood, California on May 15. Taylor Swift stopped by a gym for a workout in New York City on May 15. Calvin Klein Collection and euphoria Calvin Klein celebrated Women in Film on May 15 during the 67th Cannes Film Festival with Rooney Mara, Naomi Watts, Lupita Nyong'o and Julianne Moore. Blake Lively attended the "Mr.Turner" premiere at the 67th Annual Cannes Film Festival on May 15. Lady Gaga posed for pictures with fans as she and her boyfriend Taylor Kinney left their apartment building in New York City on May 15. Amal Alamuddin, fiancee of George Clooney, grabbed a taxi to the American Embassy in London, England on May 15. Khloe Kardashian departed on a flight at LAX in Los Angeles, California on May 14. Katie Holmes spotted at the Winter WONKA™-land Pop-Up in downtown NYC on May 15. Kendall Jenner attended the opening ceremony and "Grace of Monaco" premiere at the 67th Annual Cannes Film Festival on May 14. Blake Lively attended the opening ceremony and "Grace of Monaco" premiere at the 67th Annual Cannes Film Festival on May 14. James Franco stopped by NBC Studios for an appearance on "The Today Show" on May 14. Peter Dinklage was enthusiastic at the "X-Men: Days Of Future Past" premiere in Beijing, China on May 13. Cameron Diaz enjoyed a hike with a friend at TreePeople Park in Beverly Hills, California on May 13. Nicole Kidman signed autographs after appearing on Le Grand Journal in Cannes, France on May 13. George Clooney's fiancee Amal Alamuddin flashed her engagement ring in London on May 13. Jennifer Lopez spotted out and about in New York City on May 12. Julia Roberts attended the New York premiere of "The Normal Heart'" at Ziegfeld Theater on May 12. Sir Ian McKellen attended the UK Premiere of "X-Men: Days of Future Past" at Odeon Leicester Square on May 12. Diane Keaton at the launch of the L`Oreal Paris `It`s That Worth It` melanoma awareness campaign in NYC on May 12. Angelina Jolie stepped out of her car on the Upper East Side on May 12 in New York City. Pregnant singer Christina Aguilera and fiance Matthew Rutler were spotted out for coffee on Mother's Day with her son Max in Los Angeles, California on May 11. Jennifer Lawrence wowed at the New York premiere "X-Men: Days Of Future Past" at the Jacob Javits Center in New York City on May 10. Jenna Dewan spotted in a little black dress while leaving her hotel on May 9 in New York City. Chrissy Teigen and Curtis Granderson attended the The New National Campaign "Pledge. Drink. Win," encouraging kids to drink more water. Photo taken at MLB Fan Cave on May 8 in New York City. Jimmy Kimmel took a break from his hosting duties on May 8 to hang out with Penelope Cruz, who brought the Nespresso VertuoLine coffee to the show. "Game of Thrones" star Maisie Williams arrived at the BBC Radio 1 studios in London on May 8. Lucy Hale looked gym-ready in LA on May 8. Jennifer Lopez was snapped backstage at "American Idol" looking red hot on May 7. Mariah Carey arrived to the "Late Show with David Letterman" in NYC on May 7. Supermodel Gisele Bundchen attended the 2014 Rainforest Alliance Gala at Museum of Natural History in NYC on May 7. Jake Gyllenhaal accompanied his reported girlfriend Alyssa Miller while she walked her dog in New York City on May 7. Blake Lively stepped out of a downtown New York City hotel wearing a shiny flowered trench coat on May 7. Benedict Cumberbatch and Dakota Johnson were spotted out and about in New York City on May 6. Jennifer Aniston photographed on the set of "Cake" in Sylmar, California on May 6. Sean Penn and Charlize Theron were spotted holding hands while out and about in New York City on May 6. Elle Fanning and Angelina Jolie attend a photocall for the film "Maleficent" at Hotel Bristol on May 6 in Paris. Jessica Biel checked her messages while wearing an Atlas Bangle and Pendant by Tiffany & Co. while in Copenhagen on May 6. Sarah Jessica Parker attended the "Charles James: Beyond Fashion" Costume Institute Gala at the Metropolitan Museum of Art in New York City on May 5. Emma Stone attended "Charles James: Beyond Fashion" Costume Institute Gala at the Metropolitan Museum of Art in New York City on May 5. Blake Lively and Ryan Reynolds attended "Charles James: Beyond Fashion" Costume Institute Gala at the Metropolitan Museum of Art in New York City on May 5. Taylor Swift was seen stopping by a gym in New York City on May 4. Jessica Simpson and Eric Johnson arrived on a flight at LAX airport in Los Angeles, Calif., on May 4. Lea Michele attended the "Legends of OZ: Dorothy's Return" premiere held at the The Regency Village Theater in Westwood, Calif., on May 3. Ellen Page was spotted leaving the gym in West Hollywood, Calif., after a workout on May 2. Jared Leto attended the 2014 iHeartRadio Music Awards held at The Shrine Auditorium on May 1 in Los Angeles, Calif. Emma Roberts went grocery shopping at Bristol Farms in West Hollywood, Calif., on May 1. Kate Bosworth rocked a fierce red dress out and about NYC on May 1. Jessica Biel was spotted jetting out of LAX solo on April 30. Kim Kardashian and Serena Williams went shopping in Paris on April 30. Sarah Jessica Parker attended the 2014 AOL NewFronts at Duggal Greenhouse in NYC on April 29. Blake Lively filmed a scene for "The Age of Adaline" in Vancouver, Canada on April 28. Brooklyn Decker and Andy Roddick left their shoes at home for TOMS One Day Without Shoes event held to raise awareness about children’s health and education needs. Jonah Hill had some fun with fro-yo in NYC on April 28. Tori Spelling headed out of her LA home on April 28, amid reports that her husband Dean McDermott cheated on her which landed her in the hospital for nearly a week. Rihanna attended Roc Nation Sports' 1-year anniversary luncheon at TAO in NYC on April 28. Michael Douglas and Catherine Zeta-Jones attended the 41st Annual Chaplin Award Gala at Avery Fisher Hall at Lincoln Center, NYC, on April 28. Naomi Watts did some spring shopping at the farmers market in Brentwood, Calif. on April 27. Pregnant Kendra Wilkinson attended her son's soccer game at Woodland Hills, Calif., on April 27. Dianna Agron and Carey Mulligan shared a laugh while strolling through West Hollywood on April 26. Gwen Stefani and her family boarded their private jet in California on April 26. Emma Stone sizzled at "The Amazing Spider-Man 2" premiere at the Ziegfeld Theater in NYC on April 24. Kate Upton, Cameron Diaz and Leslie Mann attended The Cinema Society & Bobbi Brown with InStyle screening of "The Other Woman" at The Paley Center for Media in NYC on April 24. Khloe Kardashian did some shopping with her mother Kris in West Hollywood on April 24. Newlywed Jodie Foster grabbed a smoothie in LA on April 24. Emma Stone stopped by the "Good Morning America" studios in NYC on April 24, with her "The Amazing Spider-Man 2" co-stars Andrew Garfield and Jamie Foxx. Tori Spelling's husband Dean McDermott had a heated exchange with a paparazzo in LA on April 23 Jessica Alba landed in NYC on April 23. Rumer Willis attended the 5th Annual Elle Women in Music Celebration presented by CUSP by Neiman Marcus in Hollywood, Calif. Hosted by Elle Editor-in-Chief Robbie Myers, the April 22 event featured performances by Sarah McLachlan, Angel Haze and Betty Who. Eddie Murphy's daughters Shayne (left) and Bria Murphy attended Elle's 5th Annual Women in Music concert celebration on April 22 in Hollywood. Jason Sudeikis shared a laugh with Rebecca Hall on set of "Tumbledown" in Boston on April 22. Taylor Swift engaged in some retail therapy in NYC on April 22. Drew Barrymore gave birth to her second child, a baby girl named Frankie Barrymore Kopelman, on April 22. Cameron Diaz attended the premiere of "The Other Woman" at the Regency Village Theatre in Westwood, Calif., on April 21 Katy Perry headed to Jimmy Kimmel's studio in Hollywood, Calif., on April 21. Sofia Vergara wowed at "Good Morning America" on April 21. Selena Gomez went on a coffee run in Studio City, Calif. on April 21. Busy Philipps and husband Marc Silverstein had some fun at Coachella on April 20. Gwen Stefani headed to her parents' house on Easter Sunday, April 20, with her husband Gavin Rossdale and three kids, Kingston, Zuma and Apollo. Tia Mowry soaked up some sun in Cabo, Mexico with her husband Cory Hardrict on April 19. Jennifer Lopez and boyfriend Casper Smart had a casual dinner at Madeo in West Hollywood on April 17. Aubrey Plaza, Jason Ritter, Dade Constantine, and Kayla Dobson attended the "About Alex" premiere afterparty during the Tribeca Film Festival at Kutsher's Tribeca in NYC, April 17. Pregnant singer Christina Aguilera and her fiance Matthew Rutler were photographed out and about in New York City on April 17. Katie Holmes spotted out and about in New York City on April 17. "Downton Abbey" actress Jessica Brown Findlay hopped on a train at Manchester Piccadilly Train Station after her interview on "BBC Breakfast" on April 17 in England. Heather Graham visited NBC Studios for an appearance on "The Today Show" on April 17 in New York City. Justin Bieber and his crew, including Lil Za, were spotted out for dinner at The Grove in Los Angeles, Calif., on April 16. Georgia May Jagger, daughter of Mick Jagger, filmed a commercial for Rimmel London in London, England, on April 15. Halle Berry filmed scenes for her TV series "Extant" in Century City, Calif. on April 15. Lauren Conrad did some shopping in West Hollywood on April 15. Andrew Garfield, Emma Stone, Jamie Foxx and Dane DeHaan posed during a photocall for their film "The Amazing Spider-Man 2" prior to the German premiere of the film in Berlin on April 15. Kim Kardashian and her fiance Kanye West walked hand-in-hand as they went shopping in Givenchy, Balmain and Lanvin on April 14 in Paris, France. Channing Tatum and Jenna Dewan-Tatum joked around backstage at the 2014 MTV Movie Awards at Nokia Theatre L.A. Live in Los Angeles, Calif., on April 13. Lupita Nyong'o arrived at the 2014 MTV Movie Awards at Nokia Theatre L.A. Live in Los Angeles, Calif., on April 13. Taylor Swift and a friend were spotted out and about in New York City on April 12. Jessica Biel was all smiles as she headed out for an espresso in New York City on April 12. Leonardo DiCaprio stopped by the McDonald’s® and Stingray Desert Coachella Pool Party at the Bootsy Bellows Estate on April 12 to celebrate the launch of McDonald’s® Bacon Clubhouse Burger™ and the 2014 Corvette Stingray. Julianne Hough looked Coachella cool at The Old Navy Oasis in Indio, Calif.m on April 12. Selena Gomez and Kendall and Kylie Jenner at the first weekend of The Coachella Valley Music and Arts Festival on April 11. Emma Stone and Andrew Garfield visited BBC Radio 1 in London on April 9 to promote their new film, "The Amazing Spider-Man 2." Pregnant Kristin Cavallari was spotted running in West Hollywood on April 9. Danica McKellar left "Dancing with the Stars" practice in Hollywood on April 8. Julianne Moore was pretty in white while out in NYC April 8. Anne Hathaway wrapped herself up in a giant coat while out in NYC on April 8. Lady Gaga struck her usual unique pose outside the Roseland Ballroom on April 7. British socialite Peaches Geldof died of an "unexplained and sudden" cause on April 7. She was 25. "Dancing with the Stars" star Derek Hough took a break from rehearsal on April 7 to snack on the new Ritz crackers. Jamie Lynn Sigler was spotted purchasing a Rembrandt whitening kit at Target in New York on April 7. Faith Hill and Tim McGraw at the 2014 Academy Of Country Music Awards at the MGM Grand Hotel & Casino in Las Vegas, Nevada on April 6. Miranda Lambert and Blake Shelton at the 2014 Academy Of Country Music Awards at the MGM Grand Hotel & Casino in Las Vegas, Nevada on April 6. Carrie Underwood at the 2014 Academy Of Country Music Awards at the MGM Grand Hotel & Casino in Las Vegas, Nevada on April 6. Taylor Swift at the 2014 Academy Of Country Music Awards at the MGM Grand Hotel & Casino in Las Vegas, Nevada on April 6. Cameron Diaz made an appearance on "Wetten, dass..???" TV show on April 5 in Offenburg, Germany. Heidi Klum attended the "America's Got Talent" red carpet event at Madison Square Garden on April 4 in New York City. Nicole Richie attended Lucky FABB: Fashion and Beauty Blog Conference at the SLS Hotel in Beverly Hills on April 4. Shakira and Usher attended NBC's "The Voice" red carpet event at The Sayers Club in Hollywood on April 3. Dianna Agron was spotted out and about West Hollywood on April 3. Kiernan Shipka, Jessica Pare, Elisabeth Moss, Jon Hamm, January Jones and Christina Hendricks attended the "Mad Men" season 7 premiere at ArcLight Cinemas in Hollywood on April 2. Kate Upton, Cameron Diaz and Leslie Mann attended the U.K. premiere of their movie "The Other Woman" on April 2 in London. Lady Gaga arrived at the "Late Show with David Letterman" taping in NYC April 2. Heidi Klum joined Dr. Scholl’s to announce the DreamWalk line of insoles to help women everywhere bring their uncomfortable shoes out of the “shoe closet" in New York on April 2. Jennifer Connelly and Russell Crowe posed together before the screening of their new film "Noah" in Paris on April 1. Comedienne Amy Schumer promoted her "Finger Blaster" sketch from season 2 of the Comedy Central hit "Inside Amy Schumer" in NYC, April 1. Naomi Watts left the gym with a friend after a morning workout on April 1 in Brentwood, Calif. Hulk Hogan attended the WrestleMania 30 press conference at the Hard Rock Cafe New York on April 1. Sebastian Stan and Chris Evans posed with Captain America during The NYSE Opening Bell at New York Stock Exchange on April 1. Cobie Smulders attended The Cinema Society & Gucci Guilty screening of "Captain America: The Winter Soldier" at the TriBeCa Grand Hotel in NYC, March 31. Emma Watson wowed at the UK Premiere of "Noah" at Odeon Leicester Square on March 31 in London. Naomi Watts went for a morning jog with a friend in Brentwood, Calif., on March 31. Johnny Depp attended the "Transcendence" press conference at Grand Hyatt Hotel on March 31 in Beijing, China. Shailene Woodley, Theo James and Kate Winslet attended the European Premiere of "Divergent" at Odeon Leicester Square on March 30 in London. Lea Michele had a Marilyn moment while on the red carpet at the 27th Annual Nickelodeon Kids' Choice Awards in Los Angeles, Calif., on March 29. Taylor Swift spent some quality time with her model pal Lily Aldridge in NYC on March 28. Melanie Griffith was spotted out and about Beverly Hills on March 28. Channing Tatum and Mila Kunis attended CinemaCon at The Colosseum at Caesars Palace, Las Vegas, on March 27. Jennifer Lopez sizzled in red on "American Idol" March 27. Reese Witherspoon was spotted heading to a meeting in Beverly Hills on March 27. Taylor Swift went shopping in NYC on March 27. Kim Kardashian jetted to China from LAX on March 27. Emma Stone and Andrew Garfield attended "The Amazing Spider-Man 2" Singapore fan event at Marina Bay Sands on March 26. Pregnant actress Mila Kunis did some shopping in Beverly Hills, Calif., on March 26. Mila is reportedly expecting her first child with fiance Ashton Kutcher. Anne Hathaway visited the BBC studios in London on March 26. Kelly Rowland posed at the launch event for the new Caress Fresh Collection body washes in NYC on March 26. Crop top-clad Mindy Kaling attended PaleyFest at the Dolby Theatre in Hollywood on March 25. Steven Tyler performed at the National Music Publishers' Association "Celebration of the American Songwriter" event honoring him for his career and advocacy work, on March 25. Emma Watson rocked a tux to "'Late Show with David Letterman" in NYC on March 25. Kim Kardashian stunned in a see-through dress on "Late Night with Seth Meyers" on March 25. Gwyneth Paltrow and husband Chris Martin announced their separation on March 25. Kim Kardashian grabbed lunch in New York on March 25. Newly single Robin Thicke was all smiles in NYC March 25. Meryl Streep filmed scenes as Emmeline Pankhurst in "Suffragette" in London on March 24. Samuel L. Jackson, Scarlett Johansson and Chris Evans attended the "Captain America: The Winter Soldier" premiere at Taikoo Li Sanlitun on March 24 in Beijing, China. Stacy Keibler is pregnant! The newlywed announced that she's expecting her first child with husband Jared Pobre on March 24. Kourtney Kardashian dropped by ABC Studios for an appearance on "Good Morning America" on March 24 in New York City. Anne Hathaway enjoyed a day on the beach with friends in Miami, Fla., on March 23. Engaged couple Johnny Depp and Amber Heard were seen holding hands as they leave their hotel in New York City on March 22. Pregnant actress Olivia Wilde and a friend were spotted out for lunch in New York City on March 22. Mazel tov! Mila Kunis and Ashton Kutcher are expecting their first child together, multiple sources reported Sunday, March 23. "DWTS" star Max Chmerkovskiy slipped a Duracell Powermat backup battery onto his iPhone on March 22. After much speculation, on March 21 is was revealed that Kim Kardashian and Kanye West are Vogue's April cover stars. America Ferrera attended the premiere of "Cesar Chavez" at TCL Chinese Theatre in Hollywood on March 20. Jennifer Connelly prepped for her "Good Morning America" appearance on March 20, where she promoted her latest film "Noah." Sophia Bush met up with friends in LA on March 20. Emma Stone and Andrew Garfield posed for "The Amazing Spider-Man 2: Rise Of Electro" photocall on March 20 in Sydney, Australia. Charlize Theron posed for a bikini photo shoot in Miami on March 19. Emma Roberts was out for lunch in West Hollywood on March 19. Molly Sims added style to Wendy's #NewSaladCollection at an exclusive fashion event to celebrate their new Asian Cashew Chicken and BBQ Ranch Chicken Salads on March 19 in New York City. Nicky Hilton was spotted out and about NYC on March 19. Kate Winslet and Shailene Woodley attended the "Divergent" premiere at the Regency Bruin Theatre in LA on March 18. Emilia Clarke, Richard Plepler, Peter Dinklage and David Benioff attended the "Game of Thrones" Season 4 New York premiere at Avery Fisher Hall, Lincoln Center on March 18. Lea Michele attended the "Glee" 100th episode party held at the Chateau Marmont in LA on March 18. Reese Witherspoon headed to the salon in Beverly Hills on March 18. Kylie Minogue dropped by the BBC Radio 1 studio in London, England on March 18. Jaime King picked up designer spring fashions at T.J.Maxx in LA on March 18. Halle Berry and Goran Visnjic filmed scenes on the set of "Extant" on March 17 in LA. Blake Lively filmed a scene on the set of "The Age Of Adaline" in Vancouver, Canada on March 17. Maria Menounos left "The Wendy Williams Show" after promoting her reality TV show, "Chasing Maria Menounos," on March 17. Chic Eva Mendes was spotted in West Hollywood on March 17. Julianne Hough went for a morning jog with a friend in West Hollywood, Calif., on March 17. Nicole Scherzinger stepped out in London on March 17. The brunette beauty waved happily to cameras and wore a sheer black sweatshirt over a white singlet along with black leather pants and white pumps. Alyssa Milano grabbed a ride on her husband David Bulgliari's Can-Am Spyder on March 17 in LA. Kermit the Frog tried to ease tensions between fashion divas Miss Piggy and Joan Rivers during their recent adventure at QVC on March 16. The Muppets were at QVC to celebrate Disney's Muppets Most Wanted, in theaters nationwide March 21. LeAnn Rimes joined her husband Eddie Cibrian for his boys' baseball game in Calabasas, Calif., on March 16. Wilmer Valderrama took in live performances at The Naked Grape Wines Music Box during SXSW in Austin, Tex., on March 15. Courtney Stodden and her deep tan ate lunch in Beverly Hills on March 14. Kristin Cavallari rocked leather tights in NYC on March 14. Scarlett Johansson attended "Captain America: The Winter Soldier" premiere at the El Capitan Theatre on March 13 in Hollywood, Calif. Kim Kardashian was spotted leaving the Versace Mansion in Miami on March 13. Hilary Duff grabbed breakfast at La Conversation cafe in West Hollywood on March 13. Emma Roberts engaged in some retail therapy in Beverly Hills on March 13. Rita Ora was seen out and about in NYC on March 13. Kanye West performed at SXSW on March 12 in Austin, Texas. Stylish Reese Witherspoon attended a baby class in Brentwood, Calif. on March 12. Birthday girl Liza Minnelli stopped by the NBC studios in NYC for an appearance on "Today," March 12. Scott Disick and Kourtney Kardashian were seen out and about Miami on March 12. Kim Kardashian waved to her fans from the balcony in Miami, March 12. Pregnant Kristin Cavallari left the Sirius XM studios in NYC on March 12. Victoria's Secret model Alessandra Ambrosio showed the brand's 2014 Swim Collection on "Extra" at Universal Studios, LA, on March 11. Selena Gomez shot an Adidas commercial in NYC on March 11. Jimmy Fallon stepped out of his NYC home on March 11. Rumer Willis made a Starbucks run on March 11 in West Hollywood. Jason Sudeikis and pregnant Olivia Wilde enjoyed a romantic stroll in New York City for Olivia's birthday on March 10. Stacy Keibler gave herself a manicure using Sally Hansen Triple Shine nail color on March 10 in LA. Sean Avery took a break from "Dancing with the Stars" practice to read "Way of the SEAL" in LA on March 10. Britney Spears, her boyfriend David Lucado, her ex-husband Kevin Federline and his wife Victoria Prince watched their sons Sean and Jayden play soccer in Calabasas, Calif., on March 9. "True Detective" star Michelle Monaghan picked up food from Earthbar in West Hollywood on March 7. Aaron Paul arrived at the premiere of "Need for Speed" at TCL Chinese Theatre on March 6. Sarah Jessica Parker presented The SJP Collection at LA Nordstrom on March 6. James Franco, Leighton Meester and Chris O'Dowd attended the "Of Mice and Men" press conference at Signature Theatre, NYC, on March 6. Taylor Swift headed to the gym in LA on March 6. Ireland Baldwin and her new purple hair were spotted grocery shopping in Tarzana, Calif. on March 6. "Gossip Girl" star Jessica Szohr went shopping in LA on March 6. Lupita Nyong'o, Elizabeth Olsen, Bella Heathcote and Elle Fanning attended the Miu Miu show at Paris Fashion Week on March 5. Jennifer Hudson stopped by the Sony Music offices in London on March 5. Minka Kelly left her LA gym after a workout on March 5. Mandy Moore was spotted out and about West Hollywood on March 5. Julianne Moore was spotted on the set of "Still Alice" in NYC on March 4. Courteney Cox and boyfriend Johnny McDaid of "Snow Patrol" did some shopping in West Hollywood on March 4. Jared Leto arrived at LAX for a flight, March 4. Engaged couple Ashton Kutcher and Mila Kunis went out for dinner at the Black Market Liquor Bar in Studio City, Calif., on March 3. Jessica Biel caught a flight out of LAX Airport on her 32nd birthday March 3 in Los Angeles, Calif. Jessica is flying to Florida to meet up with her husband Justin Timberlake (who's currently on tour there) to celebrate her birthday with him. Sophia Bush appeared on Nick Lachey`s first episode as Host of VH1`s Big Morning Buzz in NYC on March 3. Jennifer Lawrence attended the 2014 Vanity Fair Oscar Party hosted by Graydon Carter on March 2 in West Hollywood, Calif. Ben Affleck and Jennifer Garner attended the 2014 Vanity Fair Oscar Party hosted by Graydon Carter on March 2 in West Hollywood, Calif. Sofia Vergara attended the 2014 Vanity Fair Oscar Party hosted by Graydon Carter on March 2 in West Hollywood, Calif. Ryan Seacrest posed with John Stamos at Mike De Luca and Dana Brunetti’s Oscar after party in the Hollywood Hills on March 2, where McDonald’s World Famous fries, Egg White Delights, Egg McMuffins, and hash browns were served. "12 Years a Slave" wins Best Picture at the Oscars, March 2. Matthew McConaughey wins Best Actor at the Oscars, March 2. Cate Blanchett wins Best Actress at the Oscars, March 2. Jared Leto wins Best Supporting Actor at the Oscars, March 2. Lupita Nyong'o wins Best Supporting Actress at the Oscars, March 2. Alfonso Cuaron wins Best Director at the Oscars, March 2. Brad Pitt and Angelina Jolie arrived at the 86th Academy Awards on March 2. Leonardo DiCaprio and Christoph Waltz arrived at the 86th Academy Awards on March 2. Meryl Streep arrived at the 86th Academy Awards on March 2. Jennifer Lawrence arrived at the 86th Academy Awards on March 2 and fell, again. Kerry Washington arrived at the 86th Academy Awards on March 2. Anne Hathaway arrived at the 86th Academy Awards on March 2. Pregnant Olivia Wilde and Jason Sudeikis arrived on the red carpet for the 86th Academy Awards on March 2. Anna Kendrick arrived on the red carpet for the 86th Academy Awards on March 2. Portia de Rossi arrived on the red carpet for the 86th Academy Awards on March 2. Kristen Bell arrived on the red carpet for the 86th Academy Awards on March 2. "12 Years a Slave" director Steve McQueen arrived on the red carpet for the 86th Academy Awards on March 2. Camila Alves toasted her glam team with a Bailey's Glamour Shot before hitting the Oscars red carpet on March 2. Sarah Michelle Gellar, Matthew McConaughey and Director Steve McQueen talked about creating a brighter future on the Yellow Carpet presented by Unilever Project Sunlight during the 2014 Film Independent Spirit Awards on March 1. Kanye West attended the Celine show at Paris Fashion Week on March 2. Kendall Jenner went sightseeing at the 'Musee du Louvre' in Paris, France on March 1. Pregnant Kendra Wilkinson wowed at the 5th Annual QVC Red Carpet Style held at the The Four Season Hotel in Beverly Hills, Calif., on Feb. 28. Julianne Hough met a friend for lunch at Joan's on Third in Los Angeles, Calif., on Feb. 28. Leslie Mann and Nikolaj Coster-Waldau filmed scenes for the "The Other Woman" in LA Feb. 27. Hilary Duff left her LA gym on Feb. 27. Jessica Alba attended the Nina Ricci show at Paris Fashion Week Feb. 27. Rihanna attended the Lanvin show at Paris Fashion Week Feb. 27. Dwyane Wade lunched with fiancee Gabrielle Union in Miami on Feb. 26. Amanda Seyfried went grocery shopping at Trader Joe's in LA on Feb. 26. Amy Adams left the gym in West Hollywood on Feb. 26. Emma Roberts and fiance Evan Peters made their way through the airport in Paris, France, on Feb. 26. Olivia Palermo and fiance Johannes Huebl arrived at the H&M; Paris fashion show on Feb. 26. Sofia Vergara and fiance Nick Loeb touched down at LAX on Feb. 26. Executive producer Joel Fields, Keri Russell, and writer Joe Weisberg attended "The Americans" Season 2 premiere after-party at The Plaza Hotel in NYC, Feb. 24. Ashley Greene attended Oakley's "Disruptive by Design" global campaign launch event at RED Studios in LA on Feb. 24. Amy Poehler and Vice President Joe Biden were guests on Seth Meyers' first "Late Night" episode Feb. 24. Michelle Dockery, Lupita Nyong'o, Liam Neeson and Julianne Moore attended the premiere of their film "Non-Stop" at the Regency Village Theatre in LA on Feb. 24. Reese Witherspoon headed out for a cup of coffee in NYC on Feb. 24. Katie Holmes was seen out for lunch with a mystery man in NYC on Feb. 24. Pregnant actress Olivia Wilde left her pilates class at Harmony Studios on Feb. 24 in West Hollywood, Calif. Giuliana Rancic was spotted buying Ban Total Refresh Cooling Body Cloths while on the go in LA, Feb. 24. Taylor Swift and Lorde were spotted out shopping at Free People in Brentwood before finishing up at Rag & Bone in West Hollywood, Calif., on Feb. 23. Raquel Welch stunned at the 16th Costume Designers Guild Awards held at The Beverly Hilton Hotel on Feb. 22. Hilary Duff was all smiles in LA on Feb. 21. Sofia Vergara filmed scenes on the beach in Sydney, Australia on Feb. 21. Taylor Swift was seen leaving the gym in West Hollywood, Calif. after a workout on Feb. 21. Kim Kardashian stopped by Milk Studios in NYC to visit her fiance Kanye West as he recorded on Feb. 20. AnnaSophia Robb attended Ringling Bros. and Barnum & Bailey presents "Legends" at Barclays Center in Brooklyn, NY, on Feb. 20. "American Idol" judges Keith Urban, Jennifer Lopez, Harry Connick, Jr., host Ryan Seacrest and former judge Randy Jackson attended the show's after-party in West Hollywood on Feb. 20. Sofia Vergara and Sarah Hyland filmed a scene for "Modern Family" in Sydney, Australia, on Feb. 20. Kanye West protected himself from the cold in NYC on Feb. 20. Padma Lakshmi stepped out in SoHo, NYC, on Feb. 20. Beyonce performed her hit song "XO" at the 2014 Brit Awards at the 02 Arena in London, England on Feb. 19. Swimsuit model Anne V rocked a black bikini at the beach in Miami Feb. 19. Ashley Greene worked on her fitness at Tracy Anderson Studios in Studio City, Calif. on Feb. 19. Dakota Fanning and boyfriend Jamie Strachan walked through New York City's East Village on Feb. 19. Keri Russell and Matthew Rhys filmed scenes for their series "The Americans" in Queens, NY, on Feb. 19. Hilary Duff hit the gym in West Hollywood, Calif. on Feb. 19. Selena Gomez grabbed coffee with a friend in Studio City, Calif. on Feb. 19. Jesse Tyler Ferguson and Eric Stonestreet filmed scenes for "Modern Family" in Sydney, Australia on Feb. 18. Lady Gaga was spotted heading to "The Tonight Show Starring Jimmy Fallon" on Feb. 18 in NYC. Reese Witherspoon looked lovely at lunch in Brentwood, Calif., Feb. 18. Ellen Page, who came out as gay last week to much support from her Hollywood peers, was seen heading out of LAX on Feb. 18. Debra Messing arrived at the "Today" show studio in NYC on Feb. 18. "The Tonight Show Starring Jimmy Fallon" premiered Feb. 17 with guest Will Smith. Amber Heard made an appearance on "Late Show with David Letterman" in NYC, Feb. 17. Kourtney and Khloe Kardashian touched down at LAX on Feb. 17. Angelina Jolie and Brad Pitt attended the BAFTA Awards held at The Royal Opera House on Feb. 16 in London, England. Lupita Nyong'o attended the BAFTA Awards held at The Royal Opera House on Feb. 16 in London, England. Leonardo DiCaprio attended the BAFTA Awards held at The Royal Opera House on Feb. 16 in London, England. Dakota Johnson went on a run on the set of "Fifty Shades of Grey" in Vancouver, Feb. 14. Taylor Swift headed to dance class in LA on Feb. 14. The 50th Anniversary cover of Sports Illustrated Swimsuit Issue was unveiled Feb. 13 and it features not one but three bikini'd babes: Chrissy Teigen, Lily Aldridge and Nina Agdal. Mel Gibson took his on again-off again girlfriend Nadia Lanfranconi to a romantic pre-Valentine's Day dinner at Wolfgang's Steakhouse in Beverly Hills on Feb. 13. Kendall Jenner walked the runway at the Marc Jacobs fashion show during Mercedes-Benz Fashion Week Fall 2014 at the New York State Armory on Feb. 13. Vogue Editor-in-Chief Anna Wintour, and actresses Lupita Nyong'o and Naomi Watts attended the Calvin Klein Collection fashion show during Mercedes-Benz Fashion Week Fall 2014 at Spring Studios on Feb. 13. Anna Kendrick attended the J. Mendel fashion show during Mercedes-Benz Fashion Week Fall 2014 at The Theatre at Lincoln Center in NYC on Feb. 13. Kristen Stewart was spotted en route to a meeting in Hollywood on Feb. 13. Helen Mirren touched down at Heathrow Airport in London on Feb. 13. Taylor Swift went shopping in Hollywood on Feb. 13. Johnny Depp and his fiancee Amber Heard arrived at the premiere of "3 Days To Kill" at the Arclight Theatre on Feb. 12 in LA. Kanye West attended the "River Of Fundament" world premiere at BAM Harvey Theater on Feb. 12 in NYC. Jennifer Lopez took part in a photo shoot on a yacht in Miami, Florida on Feb. 12. Selena Gomez grabbed slurpees with some friends in LA on Feb. 12. Blake Lively posed backstage at the Michael Kors fashion show during Mercedes-Benz Fashion Week Fall 2014 at Spring Studios on Feb. 12. Malin Akerman stopped to refuel her car in West Hollywood, Calif. on Feb. 12. Anna Faris was all smiles as she left a nail salon in West Hollywood, Calif. on Feb. 12. Chrissy Teigen arrived at the Badgley Mischka show during New York Fashion Week on Feb. 11. Shirley Temple Black, iconic child star and former U.S. ambassador to Ghana and Czechoslovakia, died on Feb. 10 in California. Cause of death was not released. She was 85. Olivia Munn attended the premiere of "Robocop" on Feb. 10 in Hollywood, Calif. Katie Holmes, Rita Ora, Trudie Styler, Deborra-Lee Furness and Hugh Jackman attended the Donna Karan New York 30th Anniversary fashion show during Mercedes-Benz Fashion Week Fall 2014, on Feb. 10. Academy Award nominees, including Sandra Bullock, Amy Adams, Philippe Le Sourd, Alfonso Cuaron, Karen O, and Leonardo DiCaprio, attended the 86th Academy Awards nominee luncheon at The Beverly Hilton Hotel on Feb. 10. Jeremy Piven got ready for a ride on the Can-Am Spyder in Los Angeles, Calif. on Feb. 10. Julianne Hough rocked a new pixie haircut while departing on a flight at LAX airport in Los Angeles, Calif., on Feb. 9. Stellan Skarsgard, Bente Trier, Lars von Trier, Uma Thurman, Christian Slater and Stacy Martin attended the "Nymphomaniac Volume I" premiere during the 64th Berlinale International Film Festival on Feb. 9 in Berlin, Germany. Sophia Bush volunteered with Trio Animal Foundation to support the Barefoot Wine Soles of the Year Program on Feb. 8. Lady in white: January Jones visited the Sunset Tower Hotel in Los Angeles, Calif. on Feb. 7. Ashley Tisdale visited the the Andy LeCompte Salon in West Hollywood, Calif. on Feb. 7. Meryl Streep attended the funeral service for actor Philip Seymour Hoffman at St. Ignatius of Loyola in NYC on Feb. 7. Leonardo DiCaprio and Martin Scorsese attended the Cinema Vanguard Award to Martin Scorsese and Leonardo DiCaprio at the Arlington Theatre on Feb. 6 in Santa Barbara, Calif.

Summary: As Dos Equis beer's "most interesting man in the world," Jonathan Goldsmith's resume includes such feats as walking on fire and training canaries in the art of falconry. But now the real-life Goldsmith is adding a pretty interesting item to his own resume: tracking down and eliminating landmines, USA Today reports. Goldsmith is aiming to raise money for landmine removal in Cambodia after accidents involving them spiked this year. And his fundraising contest (which closes Oct. 8, 2014) sounds pretty interesting: The winner gets to spend a day with Goldsmith in Vermont, and the 76-year-old says it will be spent "doing some things-off-road driving and some other little events, one being falconry." Other prizes include a signed box of cigars, the Burlington Free Press reports. Another interesting tidbit about Goldsmith: He has saved a girl's life.

multi_news

en

Summarize: Richard Lee sued the city of Seattle to release photos of Cobain’s body, believing the musician to have been killed by government officials A Seattle court has ruled that photographs from the scene of Kurt Cobain’s death will remain sealed from the public. In 2014, conspiracy theorist Richard Lee had sued the city over the release of the images, believing them to support his long-held view that Cobain was killed by government officials. Cobain’s death on 5 April 1994 was ruled suicide. Court documents state that the images depict “Cobain’s body as it lay in the family residence after he was shot in the head”. The court of appeals ruled in favour of Cobain’s widow, Courtney Love, and daughter, Frances Bean Cobain, who had sought a ruling “permanently enjoining the City from disclosing, disseminating, releasing, or distributing any death-scene photographs not previously disclosed”, the Blast reports. Lee lost his original suit in 2015 and filed an appeal in 2017. Cobain’s next of kin have previously testified in the case. Love has described his death as “the most traumatic experience of my life. It left me physically distraught, and I continue to suffer emotionally from the loss of my husband to this day.” His daughter testified that she had no memory of her father, but “has had to deal with the trauma of his death her entire life”. In April 2016, Love gave a statement accusing Lee of trying to exploit Cobain’s death, which he claims to have been investigating for more than 23 years. Love claimed that Lee “stalked and harassed me, my family and my friends for many, many years … On one particular occasion, Mr Lee even filmed himself chasing a limousine for several miles that he thought I was a passenger in. Mr Lee’s actions make me fear for my safety.” 'This is the pop culture legacy business': JAM Inc manages artists after death Read more Seattle police have previously made available photographs of the gun that Cobain used to kill himself along with less explicit photographs of the death scene and its aftermath. On the 20th anniversary of Cobain’s death in 2014, detective Mike Ciesynski reviewed the case files and said he found no new information to change the police conclusion that Cobain took his own life. He did find four rolls of undeveloped film from the suicide scene. After releasing two of the images in 2014, police released 34 additional photographs taken at the scene following numerous public disclosure requests. Frances Bean Cobain‘s divorce may be settled but she has lost one prized possession to her ex: her father’s famous guitar. Kurt Cobain’s daughter settled her divorce to ex Isaiah Silva on Monday in court documents obtained by PEOPLE. The 25-year-old married Silva in 2014 and filed for divorce in March 2016. In November, a judge declared the daughter of the Nirvana frontman a single woman despite the fact that the two were still dividing their assets. One point of contention between them: the iconic Martin guitar Cobain played during his MTV Unplugged performance in November 1993. Cobain died in the following year in April of suicide. Silva claimed the model had given him the guitar as a present, while she denied ever giving it to him. In November, The Blast reported the guitar is worth millions of dollars. Kurt Cobain in 1993's MTV Unplugged Despite the artist’s request for the judge to keep the guitar in her family, Silva will get to keep it. In documents filed at the time, Cobain stated Silva should not be entitled to any money from her father’s estate, which is valued at $450 million. The filing asked that all premarital assets including Cobain’s inheritance be awarded to her as separate property. RELATED: Frances Bean Cobain Is Officially Single but Is Fighting Ex for Guitar Belonging to Father Kurt Get push notifications with news, features and more. Frances Bean Cobain and Isaiah Silva Splash The artist has been dating Matthew Cook since last fall, sharing Instagram photos and videos of him during their time together. In April, she shared her first original song on the anniversary of her father’s death in a snippet on Instagram. “I think I saw you when I was small/ I think I found you/ A penny for your good thoughts,” she sings in a video. “I think I found you/ Jesus hangs in your place on the cross/ All these hinges become unscrewed / Heaven knows it was a cage on earth.” Cobain also writes to a user in the comments section that the full song features “a very freaky deeky line that goes ‘stable sable sold her heart/ no one asks her why she hides it in a casket in her house in a box/ find a fiend who reigns supreme in may/ fast enough for blooming buds to lay their eggs.’” Frances Bean Cobain Stefanie Keenan/Getty When asked what the Nirvana frontman would think of her foray into songwriting, Frances offered an emotional and thoughtful response. “I don’t have an answer for that because I don’t want to speak on someone else’s behalf,” she told E! News on the red carpet of the Daily Front Row Fashion Awards in Los Angeles in April. “I would hope that he would be proud of the human being I am even if he didn’t like the art I am putting out. That’s all I would ask of anyone in my life.” Want to keep up with the latest from PEOPLE? Sign up for our daily newsletter to get our best stories of the day delivered straight to your inbox. RELATED VIDEO: Frances Bean Cobain Reveals Her Private Battle with Addiction as She Celebrates 2 Years Sober The multi-talented artist also expanded on where she plans to place her creative efforts. “With regards to music, I don’t want to pigeonhole myself and say I am a musician or a visual artist because I feel like it’s all-encompassing and I feel like every bit of my art is related to the other,” she explained. “So do I want to pursue my music further and see it come to fruition and see something further and see something palpable? Absolutely.” Kurt Cobain and Nirvana performed on 'MTV Unplugged' in November 1993. (Photo: MTV) SEATTLE — Photographs from the scene of Kurt Cobain’s death will not be released to the public, the Washington State Court of Appeals ruled Tuesday. The court ruled the photographs are exempt from Washington state’s Public Records Act and releasing the photos would “violate the Cobain family’s due process rights under the 14th Amendment of the United States Constitution.” Cobain’s widow, Courtney Love, and his daughter who was a toddler at the time of his death, Frances Bean Cobain, filed testimonies to keep the photos from being made public. The ruling comes after Seattle journalist Richard Lee appealed the case’s dismissal. Lee has pursued the release of 55 photos in an attempt to prove Cobain did not die from suicide in 1994 but was murdered. He has been testing the limits of public records laws for years, and in the process has angered Cobain's family. “As both a father and an advocate for victims’ rights, I’m relieved the Court upheld that death-scene images are not appropriate for disclosure,” Seattle City Attorney Pete Holmes said in a statement. “After a family member endures the tragedy of losing a loved one, we have a moral obligation to protect their privacy. No one should worry whether they’ll happen upon photos of a family member’s body as they scroll through their social media feed.” Cobain, the frontman for Seattle grunge band Nirvana, was found dead of a self-inflicted gunshot wound to the head in his Seattle home in 1994. Love and her daughter wrote a letter to the judge in the trial court seeking to block the release of the photos. Frances Bean Cobain wrote that she already faces harassment from fans "obsessed" with her father and fears that could get worse. "I have had to cope with many personal issues because of my father's death," Cobain wrote. "Coping with even the possibility that those photographs could be made public is very difficult. Further sensationalizing it through the release of these pictures would cause us indescribable pain." “As a member of Washington State’s Sunshine Committee, I regularly advocate to open more records for public access, but out of respect to family members, I continue to believe releasing images of a person’s scene of death is out-of-bounds," Holmes said. CLOSE The detective on the case of Kurt Cobain's death back in 1994 recently pulled out the file and found four rolls of undeveloped film from inside musician's house the day he died. VPC Contributing: Ted Land, KING-TV, Seattle, Associated Press. Follow KING-TV on Twitter: @KING5Seattle Read or Share this story: https://usat.ly/2IlUBlT

Summary: A conspiracy theorist who thinks the government killed Kurt Cobain has once again failed to persuade judges that graphic photos of the Nirvana frontman's death should be released. A Seattle court of appeals ruled against Richard Lee this week, saying the photos were exempt from Washington state's Public Records Act and releasing them would violate the Cobain family's due process rights, USA Today reports. Cobain's April 5, 1994 death was ruled a suicide. The singer killed himself with a shotgun, and legal papers state the photos show the body "after he was shot in the head." An earlier lawsuit from Lee seeking the photos was also tossed out, though police released previously unseen photos from the scene in 2014. Release of the photos had been firmly opposed by widow Courtney Love and daughter Frances Bean Cobain, Guardian reports. In a letter to the judge, Frances, who was a toddler when her father killed himself, wrote that releasing the scene-of-death photos would cause "indescribable pain." She also said she feared such a release would worsen the harassment from obsessed fans she already faces. The 25-year-old is the singer's only child. According to court papers filed Monday as part of a divorce settlement, she has agreed to let ex-husband Isaiah Silva keep the iconic guitar Kurt Cobain played in a Nov. 1993 Unplugged appearance People reports. She had been fighting her ex in court to keep the Martin D-18E guitar, which Silva claimed had been given to him as a wedding present.

multi_news

en

Summarize: Democrats on the Senate Judiciary Committee have privately requested to view a Brett Kavanaugh-related document in possession of the panel’s top Democrat, Dianne Feinstein, but the senior California senator has so far refused, according to multiple sources familiar with the situation. The specific content of the document, which is a letter from a California constituent, is unclear, but Feinstein’s refusal to share the letter has created tension on the committee, particularly after Feinstein largely took a back seat to her more junior colleagues last week, as they took over Kavanaugh’s confirmation hearings with protests around access to documents. The letter took a circuitous route to Feinstein, the top-ranking Democrat on the Judiciary Committee. It purportedly describes an incident that was relayed to someone affiliated with Stanford University, who authored the letter* and sent it to Rep. Anna Eshoo, a Democrat who represents the area. Different sources provided different accounts of the contents of the letter, and some of the sources said they themselves had heard different versions, but the one consistent theme was that it describes an incident involving Kavanaugh and a woman while they were in high school. Kept hidden, the letter is beginning to take on a life of its own. Eshoo passed the letter to her fellow Californian, Feinstein. Word began leaking out on the Hill about it, and Feinstein was approached by Democrats on the committee, but she rebuffed them, Democratic sources said. Feinstein’s fellow senators want their own opportunity to gauge whether or not the letter should be made public, rather than leaving it to Feinstein to make that call unilaterally. The sources were not authorized to speak on the record, and said that no senators on the committee, other than Feinstein, have so far been able to view the letter. Sen. Dianne Feinstein, D-Calif., on Thursday threw a cryptic curveball at Brett Kavanaugh, insinuating the Supreme Court nominee could be guilty of a crime even as Democrats on the Senate Judiciary Committee seek to delay his confirmation. The vague accusation comes after the Senate Judiciary Committee already grilled Kavanaugh and other witnesses and prepares to vote on sending his nomination to the full Senate. The White House blasted the ambiguous charge as a last minute gambit. “I have received information from an individual concerning the nomination of Brett Kavanaugh to the Supreme Court,” Feinstein said in her surprise statement. “That individual strongly requested confidentiality, declined to come forward or press the matter further, and I have honored that decision. I have, however, referred the matter to federal investigative authorities.” A spokesperson for Feinstein declined Fox News’ request to elaborate on the lawmaker’s statement, but there has been much speculation that she is referring to a secret letter that has been the subject of intrigue on Capitol Hill over the last few days. A source familiar with the confirmation proceedings told Fox News that Feinstein received the letter back in July, but did not make its existence known publicly until Thursday. According to a report by The Intercept, the letter was relayed to lawmakers by an individual affiliated with Stanford University and concerns an incident involving the 53-year-old Kavanaugh and a woman while they were in high school. According to two officials who spoke anonymously with the New York Times, the incident involved possible sexual misconduct between Kavanaugh and the woman. The letter reportedly was given to Feinstein by Rep. Anna Eshoo, D-Calif., but has not been publicly disclosed by senators who have seen the document. Sen. Dick Durbin, D-Ill., said that the letter in question has been referred to the FBI for investigation. Two sources familiar with the matter tell Fox News that Feinstein has had possession of the letter regarding Kavanaugh since July. Feinstein met privately with Kavanaugh on August 20 and also questioned him repeatedly in open and closed session during the Judiciary Committee hearings on his nomination last week. There is no indication that the matter came up in either the private meeting or the closed committee session. The FBI conducts background checks on all major government appointees, including Supreme Court nominees. “Upon receipt of the information on the night of September 12, we included it as part of Judge Kavanaugh’s background file, as per the standard process,” the FBI said in a statement. Fox News has learned that the White House would have to request that the bureau follow up on the letter for the matter to be investigated further. It was not clear whether the White House had done so as of Thursday evening. The woman referenced in the letter has yet to be identified, but is being represented by Debra Katz, a whistleblower attorney who works with #MeToo survivors, according to The Intercept. Despite the turmoil over the letter, a spokesperson for Senate Judiciary Committee Chairman Chuck Grassley, R-Iowa, said there is no plan to delay Kavanaugh’s confirmation. Grassley set the panel's vote on Kavanaugh for Sept. 20 and Republicans hope to confirm Kavanaugh by the start of the new court session Oct. 1. “Sen. Grassley is aware of Senator Feinstein’s referral,” Grassley’s communications director Taylor Foy said in a statement. “At this time, he has not seen the letter in question, and is respecting the request for confidentiality. There’s no plan to change the committee’s consideration of Judge Kavanaugh’s nomination.” The White House attacked Feinstein's statment as an 11th hour attack on the nominee. “Throughout his confirmation process, Judge Kavanaugh has had 65 meetings with senators—including with Senator Feinstein—sat through over 30 hours of testimony, addressed over 2,000 questions in a public setting and additional questions in a confidential session," White House spokesperson Kerri Kupec said in a statement. "Not until the eve of his confirmation has Sen. Feinstein or anyone raised the specter of new ‘information’ about him." Fox News' Jake Gibson, Mike Emanuel and John Roberts contributed to this report. Senate Judiciary Committee Chairman Chuck Grassley, R-Iowa, left, accompanied by Sen. Dianne Feinstein, D-Calif., the ranking member, center, speaks with Sen. Patrick Leahy, D-Vt., right, during a Senate... (Associated Press) Senate Judiciary Committee Chairman Chuck Grassley, R-Iowa, left, accompanied by Sen. Dianne Feinstein, D-Calif., the ranking member, center, speaks with Sen. Patrick Leahy, D-Vt., right, during a Senate Judiciary Committee markup meeting on Capitol Hill, Thursday, Sept. 13, 2018, in Washington. The... (Associated Press) Senate Judiciary Committee Chairman Chuck Grassley, R-Iowa, left, accompanied by Sen. Dianne Feinstein, D-Calif., the ranking member, center, speaks with Sen. Patrick Leahy, D-Vt., right, during a Senate Judiciary Committee markup meeting on Capitol Hill, Thursday, Sept. 13, 2018, in Washington. The... (Associated Press) Senate Judiciary Committee Chairman Chuck Grassley, R-Iowa, left, accompanied by Sen. Dianne Feinstein, D-Calif., the ranking member, center, speaks with Sen. Patrick Leahy, D-Vt., right, during a Senate... (Associated Press) WASHINGTON (AP) — The Latest on Supreme Court nominee Brett Kavanaugh (all times local): 1:20 p.m. Sen. Dianne Feinstein (FYN'-styn) says she's notified federal investigators about information she received concerning Supreme Court nominee Brett Kavanaugh. The California Democrat says in a statement that she "received information from an individual concerning the nomination." Feinstein isn't saying who that person is or describing the information in any way. She says the person "strongly requested confidentiality, declined to come forward or press the matter further, and I have honored that decision." Feinstein — who's on the Senate Judiciary Committee, which just finished confirmation hearings for Kavanagh — says she has "referred the matter to federal investigative authorities." Another committee Democrat, Illinois Sen. Dick Durbin, says the matter has been referred to the FBI. Republicans are pushing to confirm Kavanaugh to the court by Oct. 1, when the 2018-19 term begins. ___ 11:05 a.m. Republicans have rejected an effort to subpoena documents on Supreme Court nominee Brett Kavanaugh's job as staff secretary in the George W. Bush White House. Democrats on the Judiciary Committee say the documents are needed to vet the judge's record. The panel's top Democrat, Sen. Dianne Feinstein (FYN'-styn) of California, said Thursday senators "should be able to see this record." Feinstein says it makes her wonder "what in Judge Kavanaugh's records are Republicans hiding?" Republicans on the panel rejected the motion on a party line vote. The Republicans declined to pursue Kavanaugh's staff secretary documents as too cumbersome, focusing instead on his White House counsel's work. President Donald Trump's nominee for the court is the first with a lengthy email trail. Democrats say the records they've seen are insufficient. ___ 10:50 a.m. Democrats have tried to adjourn the Senate Judiciary Committee's hearing to consider Supreme Court nominee Brett Kavanaugh, but Republicans have rejected the motion. Sen. Richard Blumenthal of Connecticut protested as soon as the hearing gaveled opened Thursday. He says the nomination will be "tainted" and "stained" by the unusual process for vetting President Donald Trump's nominee. Democrats say Republicans, who have the majority, are rushing the proceedings. Blumenthal says, "We lack the time. We lack the documents." He calls it a "badly broken process." Republican Chairman Chuck Grassley of Iowa says he already agreed to hold over the vote on Kavanaugh for one week. The panel will vote next week on whether to recommend Kavanaugh for confirmation. Republicans hope to confirm him to the court by Oct. 1. ___ 12:40 a.m. Supreme Court nominee Brett Kavanaugh says he would have shaken the hand of a man whose daughter was killed in a Florida high school shooting if he had known who the man was. A photo of Kavanaugh appearing to refuse to shake Fred Guttenberg's outstretched hand last week went viral. Kavanaugh told the Senate Judiciary Committee he assumed the man who approached him was a protester. He says if he had known who Guttenberg was, "I would have shaken his hand, talked to him, and expressed my sympathy. And I would have listened to him." Kavanagh's explanation was part of a 263-page response late Wednesday to some 1,287 written questions from senators. The Senate Judiciary Committee is set to meet Thursday to consider Kavanaugh's confirmation.

Summary: Sen. Dianne Feinstein says she's notified federal investigators about information she received concerning Supreme Court nominee Brett Kavanaugh, the AP reports. The California Democrat says in a statement that she "received information from an individual concerning the nomination." Feinstein isn't saying who that person is or describing the information in any way. She says the person "strongly requested confidentiality, declined to come forward or press the matter further, and I have honored that decision." Feinstein-the top-ranking Democrat on the Senate Judiciary Committee, which just finished confirmation hearings for Kavanagh-says she has "referred the matter to federal investigative authorities." Another committee Democrat, Illinois Sen. Dick Durbin, says the matter has been referred to the FBI. Fox News refers to the whole thing as "cryptic." Confirmed details are scarce, but the Intercept says the information Feinstein is referring to came to her via a letter that was first sent to US Rep. Anna Eshoo from someone affiliated with Stanford University.

multi_news

en

Summarize: FOR IMMEDIATE RELEASE – April 13, 2016 (Bowmanville, ON) – The Ontario Society for the Prevention of Cruelty to Animals has formally filed animal cruelty charges against Michael Hackenberger, the owner of the Bowmanville Zoo. The Ontario SPCA was provided with footage of Mr. Hackenberger and Uno, a male Siberian tiger, during a training session where Hackenberger used a whip to strike the animal repeatedly. The Ontario SPCA immediately started an investigation after reviewing the footage. Today, the Ontario SPCA served Mr. Hackenberger a summons related to five charges of animal cruelty under the Ontario Society for the Prevention of Cruelty to Animals Act. The charges are listed as: one charge of causing an animal to be in distress by striking the animal with a whip handle; one charge of causing an animal to be in distress by repeatedly striking the animal with a whip; one charge of causing an animal to be in distress by striking the animal in the face with a whip; one charge of causing an animal distress by pushing his thumb into the animal’s eye and; one charge being the person who fails to comply with the prescribed standards of care for such an animal and thereby failed to provide the necessary care for its general welfare as required by the Ontario Regulation 60/90 Section 2(3). “The videos of Mr. Hackenberger interacting with Uno, the Siberian tiger, provides a basis on which to lay charges,” says Senior Inspector Jennifer Bluhm, Ontario SPCA. “Animal cruelty is a serious offense. Our investigative unit has spent significant time reviewing the facility and interviewing all involved. Our priority is always the health and welfare of the animals.” The Ontario SPCA will be doing another inspection of the Bowmanville Zoo and following that; all of the animals at the zoo will continue to be closely monitored by the Ontario SPCA. The Ontario SPCA Act gives authority to the Society to investigate reports of animal cruelty and to inspect facilities to ensure the standards of care are being met at facilities were animals are being kept for the purpose of animal exhibition, entertainment, boarding, hire or sales, legislation does not grant the Society jurisdiction over the operations of a zoo. The Ontario SPCA does not have the authority to grant nor to revoke anyone the right to own or care for an animal. -- 30 -- Media contact: Alison Cross Ontario SPCA 905-853-2108 Ontario SPCA and Humane Society: Protecting animals since 1873, Ontario SPCA is Ontario's Animal Welfare organization. A registered charity comprised of over 50 communities. Since 1919, when Ontario's first Animal Welfare legislation was proclaimed, the Ontario SPCA, with the help of its Communities, has been entrusted to maintain and enforce Animal Welfare legislation. The Act provides Ontario SPCA Agents and Inspectors with police powers to do so. Ontario SPCA provides leadership in animal welfare innovations including introducing high-volume spay/neuter services to Ontario and opening the Provincial Education and Animal Centre. OntarioSPCA.ca Adopt • Learn • Volunteer • Donate Charitable Business Number 88969 1044 RR0002 BOWMANVILLE -- Bowmanville Zoo owner Michael Hackenberger has been charged with five counts of animal cruelty four months after a video was made public of him training a Siberian tiger. Related Stories Investigation and protests whipped... The Ontario Society for the Prevention of Cruelty to Animals formally filed animal cruelty charges against Mr. Hackenberger on April 12. The Ontario SPCA began its investigation after viewing footage of Mr. Hackenberger and Uno, a male Siberian tiger, during a training session where Mr. Hackenberger used a whip to strike the animal repeatedly, says the Ontario SPCA. The video was made public last December. PETA’s YouTube channel The five charges of animal cruelty under the Ontario Society for the Prevention of Cruelty to Animals Act are: causing an animal to be in distress by striking the animal with a whip handle; causing an animal distress by repeatedly striking the animal with a whip; causing an animal to be in distress by striking the animal in the face with a whip; causing an animal distress by pushing his thumb into the animal’s eye: and failing to comply with the prescribed standards of care for such an animal. In a video rebuttal posted online by the zoo after the video was made public, Mr. Hackenberger said he could be accused of viciously whipping the ground or the air, but not the tiger. He said he hit the animal three times, once in the paw when it was reaching to claw another trainer. Bowmanville zoo YouTube channel “The videos of Mr. Hackenberger interacting with Uno, the Siberian tiger, provides a basis on which to lay charges,” says Ontario SPCA senior inspector Jennifer Bluhm. “Animal cruelty is a serious offense. Our investigative unit has spent significant time reviewing the facility and interviewing all involved. Our priority is always the health and welfare of the animals.” The Ontario SPCA will be doing another inspection of the Bowmanville Zoo and following that, all of the animals at the zoo will continue to be closely monitored. These crawls are part of an effort to archive pages as they are created and archive the pages that they refer to. That way, as the pages that are referenced are changed or taken from the web, a link to the version that was live when the page was written will be preserved.Then the Internet Archive hopes that references to these archived pages will be put in place of a link that would be otherwise be broken, or a companion link to allow people to see what was originally intended by a page's authors.The goal is to fix all broken links on the web. Crawls of supported "No More 404" sites. These crawls are part of an effort to archive pages as they are created and archive the pages that they refer to. That way, as the pages that are referenced are changed or taken from the web, a link to the version that was live when the page was written will be preserved.Then the Internet Archive hopes that references to these archived pages will be put in place of a link that would be otherwise be broken, or a companion link to allow people to see what was originally intended by a page's authors.The goal is to fix all broken links on the web. Crawls of supported "No More 404" sites. The Canadian criminal charges follow the 'Life of Pi' tiger trainer being allegedly caught on video whipping a Siberian tiger in his care by animal rights group PETA. A trainer who has supplied animals for Hollywood productions and was earlier caught on video allegedly whipping and verbally abusing a Siberian tiger under his care has been slapped with criminal charges in Canada. The Ontario Society for the Prevention of Cruelty to Animals on Wednesday said it formally filed animal cruelty charges against Michael Hackenberger, the owner of the Bowmanville Zoo. The zoo keeper in December 2015 was shown in a video obtained by animal rights group PETA apparently swearing at and whipping the young tiger 19 times. The zoo is located roughly an hour outside of Toronto. In all, Hackenberger faces five charges of animal cruelty, including one charge of causing an animal distress by pushing his thumb into the animal’s eye, and another charge of failing to comply with prescribed standards of care for a tiger. "The videos of Mr. Hackenberger interacting with Uno, the Siberian tiger, provides a basis on which to lay charges,” senior inspector Jennifer Bluhm of the Ontario SPCA said in a statement. Hackenberger has supplied animals for a number of TV and film productions including Seth Rogen's The Interview as well as, most famously, Jonas, a Bengal tiger who featured prominently in Ang Lee's Life of Pi (although the 2012 film also used computer-generated visual effects to portray the tiger). A Hollywood Reporter investigation revealed in 2013 that another tiger on the set of Life of Pi allegedly nearly drowned during filming, according to an email from an American Humane Association monitor. PETA president Ingrid Newkirk welcomed the charges against the Hollywood animal exhibitor. "We are pleased that criminal charges have been brought against Michael Hackenberger for the abuse of a tiger used in his exploitive zoo shows, and we hope that this case will result in an end to his torment of animals made to suffer for human entertainment," she said in a statement. Hackenberger denied claims by PETA that he struck the Siberian tiger with a whip 19 times. The Ontario SPCA began probing the alleged animal abuse by Hackenberger after PETA revealed video of Hackenberger's alleged animal abuse. Play Facebook Twitter Embed Watch: Private Zoo Owner Caught Abusing Tiger 1:37 autoplay autoplay Copy this code to your website or blog A Canadian zoo owner whose Bengal tiger starred in the "Life of Pi" resigned Wednesday after he was charged with five counts of animal cruelty. The allegations first arose last December, after animal rights activists posted an undercover video online purporting to show the owner, Michael Hackenberger, whipping a tiger. In a statement posted late Wednesday to the Bowmanville Zoo’s Facebook page, Hackenberger said that while he was “not guilty of the charges” against him, “the welfare of the zoo and the animals that it serves has always been my principal concern. To this end I am standing down from the position of Director of Bowmanville Zoo until such time as this legal matter is resolved.” The Ontario Society for the Prevention of Cruelty to Animals, which has legal authority in animal welfare cases, filed the charges against Hackenberger after spending “significant time reviewing the facility and interviewing all involved,” the group said in a statement. This frame grab taken from a video shot by a PETA eyewitness purportedly shows Bowmanville Zoological Park owner Michael Hackenberger repeatedly whipping a Siberian tiger named Uno approximately during a training session. PETA Hackenberger was found to have struck the male Siberian tiger, Uno, in the face and elsewhere with a whip, the statement said, adding that he also caused “the animal distress by pushing his thumb into the animal’s eye.” In the roughly one-and-a-half-minute video, which shows footage from a training session and was posted by People for the Ethical Treatment of Animals on Dec. 23, Hackenberger can be seen apparently whipping Uno repeatedly and swearing at him. “The beauty of the paws being on the rock when you hit him, it’s like a vise,” he explains at one point. “It stings more.” He adds: “If we’d been running a videotape the whole time you were here, and you did a 45 second montage of the times I struck this animal, PETA would burn this place to the ground.” In a video response posted to the zoo’s Facebook page the next day, Hackenberger accused PETA of fabricating footage, and he disputed the group’s claim that he was “viciously” whipping the tiger, among other things. “I got him twice,” he said. “But after that, every whip of the whip you see, I do not strike the animal.” He adds that “a tiger will not lay on the ground and allow itself to be struck, as this videotape suggests. They’ll turn around and they’ll try to kill you. That’s not what we’re about.” If convicted, Hackenberger faces a maximum sentence of two years in jail, a $66,000 fine and a possible lifetime ban on owning an animal, Ontario SPCA spokeswoman Alison Cross told NBC News. On its website, the Bowmanville Zoo claims to be the oldest private zoo in North America. In addition to “The Life of Pi,” one of the zoo’s elephants appeared in the Adam Sandler movie “Billy Madison,” and two of its camels were the “The Thirteenth Warrior,” with Antonio Banderas.

Summary: The self-proclaimed oldest zoo in North America has provided animals to films starring names like Adam Sandler and Seth Rogen, but perhaps no Bowmanville Zoo animal has entered the spotlight like its Bengal tiger, which starred in the Oscar-winning Life of Pi. Now the Canadian zoo finds itself thrust in the spotlight for a less spectacular reason: Owner Michael Hackenberger has been charged with five counts of animal cruelty some four months after PETA released a 90-second video that allegedly captured him whipping a male Siberian tiger. The video (warning: language) also shows Hackenberger swearing at the tiger, named Uno. NBC News reports the Ontario Society for the Prevention of Cruelty to Animals said it brought the charges after spending "significant time reviewing the facility and interviewing all involved." The Ontario SPCA's statement outlines the charges, which include "one charge of causing an animal distress by pushing his thumb into the animal's eye." Hackenberger maintains his innocence but resigned Wednesday as director "until such time as this legal matter is resolved," per the zoo's Facebook page. Though the Hollywood Reporter says the video seems to show the tiger being whipped 19 times, a 31-minute video the zoo made in the wake of the PETA video has Hackenberger saying the vicious whipping was of the air and ground, with the whip actually striking the tiger only three times; on one occasion he says the animal was moving to claw another trainer, per local media. The maximum penalty is a 2-year jail terms and $66,000 fine.

multi_news

en

Summarize: This application claims the benefit of U.S. Provisional application Ser. No. 60/040,315, filed Feb. 14, 1997. FIELD OF THE INVENTION The present invention relates to colloidal and solid media for animal habitats, particularly invertebrates, and most particularly ants. The disclosed compositions can be used to create artificial habitats for invertebrates which are clean, clear or any selected color, and have the necessary nutrients for invertebrates to sustain life. BACKGROUND OF THE INVENTION In nature, the soil provides a habitat for a diverse group of animal life. In addition to providing shelter, the soil may serve as a carrier and a repository for moisture as well as other minerals and nutrients required by the organisms living therein. The growth and culture of animals, and particularly invertebrates, in a closed and contained structure is well known. Ants, in particular, have long been the subject of such artificial habitats, or &#34;ant farms,&#34; in which a structure having one or more clear sides is filled with soil. A colony of ants inhabits the soil, and may be observed to some degree through the clear enclosure walls. In order to maintain such an environment, it is frequently necessary to provide water and nutrients for the ant colony. Some nutrients may be provided by allowing plants to grow in the habitat. As a general rule, however, the continuous replenishment of water and nutrients is required to maintain a viable habitat. In addition, such a closed environment often provides a fertile breeding ground for a variety of unwanted micro-organisms. Mold, fungi and bacteria can thrive, and often provide a untimely demise to the ant population. In addition to the foregoing, artificial soil habitats have a general limitation that the container for the soil must be constructed with a relatively narrow depth to maximize the portion of the underground colony formed which can be observed through a side wall. The darkness and general nature of the soil impairs or prevents observation of ant activities more than a couple millimeters away from the side walls. The use of colloidal and solid artificial growth media are known and used for plants, bacteria, and other microbials. Such plant cultures typically consist of water, mineral salts, and other ingredients such as phytohormones, vitamins, one or more carbon sources, such as sucrose or glucose, and one or more growth enhancers. Conventional culture media must be maintained in a sterile environment to prevent contamination by bacteria and other unwanted organisms. The use of such media has generally been considered as unsuitable for use as a habitat for complete organisms such as ants or other invertebrates. Beyond the apparent differences between soil and such media, the requirement for continued sterility, and the ability of such media to support bacterial and fungal growth teach away from the use of of such media for an ant habitat. It is an objective of the present invention to provide a composition which may be used in lieu of soil as a habitat media for invertebrates and, in particular for ants. A further objective of the present invention is to provide a habitat for the continued sustenance of ant life and which may further provide a source of nutrients for the ants. A further objective of the present invention is to provide a composition and method for maintaining an artificial soil environment for ants in which microbial contamination is reduced or prevented throughout the useful life of the media. A further object of the present invention is to provide a soil-less habitat and nutritional media for normally soil-living animals, such as ants, which allows for the increased observation of such animals throughout the media. BRIEF SUMMARY OF THE INVENTION Published PCT application WO 96/38542 of the present inventor and another discloses compositions and methods for the prevention of microbial contamination of plant tissue culture media. In particular, such compositions and methods prevent or inhibit microbial growth in culture media for plant tissue cultures which normally require maintenance of sterile conditions. Specific chemical agents are used in the culture medium at concentrations that reduce or prevent microbial contamination but which allow for substantially normal germination of seeds or substantially normal growth of development of plants, plant organs, plant tissues or plant cells. The present invention is directed to a soil-less animal habitat medium comprising a cell or tissue culture medium incorporating a chemical agent at a concentration that effectively reduces or prevents the growth of bacteria and fungi and that allows for substantially normal survival and sustenance of a complex animal organism, such as an ant or a colony of such organisms. The habitat medium is in the form of a solid, and preferably a gel, which may be clear or colored, and which allows observation of the animals throughout the media for which it provides a habitat. In a particular contemplated embodiment, the habitat media may serve as a habitat for an ant colony, which can thrive in the media for an extended length of time without the necessity for external nutrient replacement. DETAILED DESCRIPTION OF THE INVENTION The present invention provides compositions and methods to maintain a self-sustaining ant habitat or colony by providing a soil-less environment capable of providing both a habitat and moisture and nutrition source for ants over an extended period. A chemical agent is incorporated into a cell or tissue culture medium in a concentration that is effective to reduce or prevent the growth of bacteria and fungi and that allows for substantially normal and sustained survival and nurture of the ants. The chemical agent useful in practicing the present invention preferably comprises a mixture of methylchloroisothazolinone (5-chloro-2-methyl-4-isothiazolin-3-one), methylisothiazolinone (2-methyl-4-isothiazolin-3-one), magnesium chloride and magnesium nitrate. More preferably, the chemical agent comprises a mixture of methylchloroisothiazolinone, methylisothiazolinone, magnesium chloride, magnesium nitrate, and potassium sorbate and sodium benzoate. Applicant has surprisingly discovered that a combination of a solid cell or tissue culture media in combination with these combinations of chemicals, in a particular range of concentrations, are effective as an ant habitat and sustaining ant life without the introduction of other nutrients while reducing or preventing microbial contamination for an extended duration, thus helping to ensure the survival of the ant colony raised within the media. The ants utilize the medium as both a replacement for a soil matrix in which a colony can be maintained, and as a nutrition source, providing both food and water. Thus, no additional foodstuffs needs to be provided. The relative concentrations of the individual components comprising the chemical agent may be varied to produce a mixture that is optimally effective in practicing the present invention for the ant colony. However, a preferred mixture of the components in the chemical agent comprises: methylchloroisothiazolinone in a concentration range of about 2.0 to about 2.6 g/l; methylisothiazolinone in a concentration range of about 0.6 to about 0.8 g/l; magnesium chloride in a concentration range of about 15.0 to 30 g/l. A more preferred mixture of the components in the chemical agent further comprises: potassium sorbate in a concentration range of about 15 to about 25 g/l or sodium benzoate in a concentration range of about 13 to about 27 g/l. In all cases, the components of the chemical agent are mixed to form a stock solution of chemical agent using any liquid in which the components will dissolve, but preferably in water, and most preferably in distilled or deionized water. As used in the present application, &#34;microbial contamination&#34; refers to the growth of any unwanted micro-organisms, e.g., bacteria or fungi, in a cell or tissue culture medium. As used in the present application, &#34;cell or tissue culture medium,&#34; &#34;culture medium,&#34; and &#34;medium&#34; refer to a solid substrate (including gels), having nutrients and other materials used for the growth of micro-organisms and plant and animal tissues in culture, including but not limited to such substrates in which a plant seed will germinate, a plant can be maintained or grown, an isolated plant or animal organ or animal tissue can be maintained, propagated or differentiated, or one or more isolated plant cells, plant cell aggregates or plant cell protoplasts may be maintained, propagated or differentiated and which is to be maintained in a sterile condition, i.e., substantially free of microbial contamination. The terms &#34;cell or tissue culture medium,&#34; &#34;culture medium,&#34; and &#34;medium&#34; are further intended to refer to solid media containing water and an appropriate mixture of mineral salts. The culture medium may further incorporate, in appropriate concentrations, phytohormones including, for example, auxins, cytokinins or gibberellins, vitamins, such as one or more B-vitamins, one or more carbon sources including, for example sucrose or glucose, and one or more undefined growth enhancers, such as coconut milk, as generally known in the art. For example, the mineral salts may be selected from any number of commercially available mixtures (e.g., from Sigma Chemical Co., St. Louis, Mo.). Mineral salt mixtures useful in the practice of the present invention include, for example, Hoagland&#39;s basal salt mixture, Gamborg&#39;s B-5 basal salt mixture, Heller&#39;s basal salt mixture, Murashige and Skoog basal salt mixture, and variations thereof. In addition, various macronutrients, micronutrients and vitamin components known in the art may be included in the culture media. Such a culture media may be, for example, Murashige and Skoog mineral salts and 2% (w/w) sucrose. According to the method of the present invention, the chemical agent is added to the culture medium in a concentration that will reduce or prevent the growth of bacteria or fungi, or both, and that will allow substantially normal growth of an ant or ant colony utilizing the culture media as a habitat and as a nutrition source. As used in the present invention, a chemical agent is effective at reducing or preventing microbial growth in a culture medium if addition of the chemical agent to the plant culture medium at the concentration that allows substantially normal growth or development of the ants, and reduces the amount of bacterial or fungal contamination by at least 80% compared to control media lacking the chemical agent. A preferred range of concentrations of the chemical agent in the culture medium is from about 0.02% to about 0.07% (v/v). A preferred chemical agent is the Kathon product of Rohn &amp; Haas. An ultimate concentration of methylchloroisothiazolinone in a range of 2.3 to 8.01 mg/l of culture media, methylisothiazolinone in a range of 0.6 to 2.1 mg/l, with magnesium salts, preferably Mg(NO 3 ) 2 and MgCl 2 in a range of 46-161 mg/l has been found preferable for culture of harvester ants of a species generally known and used in ant vivaria, such as Pogononyrmex Occidentalis. A gelling agent, for example, PHYTAGEL™ gellan gum (Sigma Chemical Co.), in an appropriate concentration range, for example, from 0.2% to 0.3% (w/v) for gellan gum, may be added to the culture medium. The culture medium must then be sufficiently heated, e.g., by autoclaving, to dissolve the gelling agent. The presence of the chemical agent, however, renders autoclaving for the purpose of sterilization unnecessary. After autoclaving, culture medium comprising chemical agent and the dissolved gelling agent may then be transferred to individual habitat containers of any type appropriate for an ant habitat or vivarium and the medium allowed to cool and solidify. An additional benefit of the chemical agent is that, where autoclaving is not needed to dissolve a gelling agent, heat-labile components of the culture medium such as vitamins and sugars will no longer require filter-sterilization. The culture medium so prepared may be stored for an extended period of time or used immediately. The present invention further provides an ant habitat kit comprising a chemical agent in a concentration that is effective to reduce or prevent microbial contamination for an extended duration and that allows for substantially normal ant life over that duration which kit comprises a culture or habitat container comprising a cell or tissue culture medium comprising the chemical agent, as well as a quantity of ants to be raised in the culture container. Having now generally described the invention, the same will be more readily understood through reference to the following examples which are provided by way of illustration, and are not intended to be limiting of the present invention. While this invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications. This application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features hereinbefore set forth as follows in the scope of the appended claims.

Summary: A soil-less habitat medium, especially for ants, allows the sustained viability of an ant or ant colony without the provision of additional nutrients. The medium comprises the combination of a culture medium and a chemical agent to reduce or prevent microbial contamination of the culture medium. The medium is preferably in the form of a clear or tinted gel, and allows the ants to be observed throughout the medium. The medium is of particular value for use in an ant vivaria.

big_patent

en

Write a title and summarize: Over two-thirds of the world' s population lives in regions where rabies is endemic, resulting in over 15 million people receiving multi-dose post-exposure prophylaxis (PEP) and over 55,000 deaths per year globally. A major goal in rabies virus (RABV) research is to develop a single-dose PEP that would simplify vaccination protocols, reduce costs associated with RABV prevention, and save lives. Protection against RABV infections requires virus neutralizing antibodies; however, factors influencing the development of protective RABV-specific B cell responses remain to be elucidated. Here we used a mouse model of IL-21 receptor-deficiency (IL-21R−/−) to characterize the role for IL-21 in RABV vaccine-induced immunity. IL-21R−/− mice immunized with a low dose of a live recombinant RABV-based vaccine (rRABV) produced only low levels of primary or secondary anti-RABV antibody response while wild-type mice developed potent anti-RABV antibodies. Furthermore, IL-21R−/− mice immunized with low-dose rRABV were only minimally protected against pathogenic RABV challenge, while all wild-type mice survived challenge, indicating that IL-21R signaling is required for antibody production in response to low-dose RABV-based vaccination. IL-21R−/− mice immunized with a higher dose of vaccine produced suboptimal anti-RABV primary antibody responses, but showed potent secondary antibodies and protection similar to wild-type mice upon challenge with pathogenic RABV, indicating that IL-21 is dispensable for secondary antibody responses to live RABV-based vaccines when a primary response develops. Furthermore, we show that IL-21 is dispensable for the generation of Tfh cells and memory B cells in the draining lymph nodes of immunized mice but is required for the detection of optimal GC B cells or plasma cells in the lymph node or bone marrow, respectively, in a vaccine dose-dependent manner. Collectively, our preliminary data show that IL-21 is critical for the development of optimal vaccine-induced primary but not secondary antibody responses against RABV infections. RABV is a single-stranded negative sense RNA virus of the genus lyssavirus in the Rhabdoviridae family that kills approximately 55,000 people annually. Up to 60% of rabies cases are in children, making rabies the seventh most important infectious disease in terms of years lost [1]. In Africa, a person dies of rabies every 20 minutes [2]. In China, rabies became the leading cause of infectious disease mortality in 2006, which increased by more than 27% from 2005 [3]. In the United States, cases of rabies in wildlife are detected in virtually all states and Puerto Rico (Hawaii is considered rabies-free). Except for cattle and foxes, the incidence of rabies in domesticated or wildlife remained unchanged or significantly increased in the US in 2011 compared to the five-year average for each species [4], exemplifying the difficulty in containing zoonotic viral infections even in industrialized nations. The cost associated with rabies in the US, Africa and Asia is almost $1 billion annually [5], [6] contributing to the financial burden of global health care costs. Furthermore, rabies is a NIAID Category C Priority Pathogen, indicating rabies is an emerging infectious disease with the potential for mass dissemination and harm to people [7]. Together, rabies is considered a neglected global zoonotic infectious disease that disproportionately affects children and, therefore, understanding how B cells develop in response to experimental RABV-based vaccination may help to support efforts to develop a single-dose human rabies vaccine for use in both developing and industrialized countries. A wide array of RABV variants exist, ranging from highly pathogenic strains to attenuated RABV vaccine strains such as the molecular clone SAD B19 [8]. Live attenuated RABV vaccine strains are highly immunogenic and potentially could serve as a single-dose human RABV vaccine to replace currently used multi-dose inactivated RABV-based vaccine regimens. Due to residual pathogenicity of these live virus strains, however, several “second-generation” RABV-based vaccines are under investigation in which entire genes are deleted from the RABV genome [9]–[12], or multiple pathogenic markers are genetically modified [13]. Data from these studies indicate that very safe and effective live RABV-based vaccine vectors can be generated. Despite extensive efforts to attenuate live RABV-based vaccine vectors for safety, little information is available on factors that influence the generation of effective antibodies in response to live RABV-based vaccines. Virus neutralizing antibodies (IgG but not IgM) directed against the RABV glycoprotein (G) are protective against pathogenic RABV strains [14], [15]. In the case of a replication-deficient RABV-based vaccine in which the matrix gene is deleted, VNAs are generated by T cell-independent and –dependent (extrafollicular and germinal center) mechanisms [16], suggesting multiple pathways of B cell activation and differentiation could be exploited to rationally design a single-dose RABV vaccine for use in both pre- and post-exposure settings. With respect to typical vaccine-induced antibody responses, APC-primed T cells most likely display an intermediate Tfh phenotype (i. e., “pre-Tfh cell”) characterized phenotypically as CD4+CXCR5hiPD1lo, which migrate the T and B cell border of secondary lymphoid organs and interact with their cognate antigen-primed B cells [17]. This T∶B cell interaction typically results in the Tfh cells producing optimal amounts of IL-21, and in the B cells differentiating into early short-lived extrafollicular antibody secreting cells or migrating into the follicles and forming GCs. With additional signals provided by Tfh cells in GCs, B cells mature and differentiate into long-lived plasma cells (PCs) secreting high affinity antibodies or into memory B cells. Due to the importance for PCs secreting high affinity antibodies and memory B cells in vaccine-induced immunity, the development of Tfh cells and CG B cells is critical for vaccine-induced protection against future exposures. In the context of RABV-specific vaccination in post-exposure settings, the rapid induction of extrafollicular B cell responses may also be critical to prevent infection of the CNS, especially in cases where treatment is delayed after exposure to a potentially infected animal. As such, understanding factors that generate short- and long-term anti-viral B cell responses will help design more efficacious RABV vaccines for use in humans. Cytokines present at the time of antigen exposure influence T and B cell activation and GC formation and, therefore, also affect the outcome of vaccination. IL-21 [18], [19] is a type 1 cytokine that is a member of the common γ-chain receptor family, which also includes IL-2, IL-4, IL-7, IL-9, and IL-15. It is produced primarily by activated Tfh and Th17 cells and has pleiotropic effects throughout innate and adaptive immunity [reviewed in [20]]. The role for IL-21 in regulating Tfh and B cell functions was originally identified using model antigens [21]–[23]. In addition, the role for IL-21 in immunity and protection against helminth [24], viral [25], [26] and bacterial [22] infections has been studied. IL-21 is a key mediator for the control of persistent viral infections in mouse models of LCMV [27]–[29] and hepatitis B virus [30], or in humans infected with HIV [31]–[33] or HIV in combination with the 2009 H1N1 influenza virus vaccine [34]. However, the complexity and diversity of persistent infections makes it is difficult to pinpoint the effects of IL-21 on anti-viral B cells versus CD8+ T cells, both of which can contribute the control of many chronic infections [35]. Conversely, B cells play a critical role for clearance of most acute viral infections and for the efficacy of vaccines against most vaccine-preventable diseases. Clearance of RABV infections relies strictly on B cell-mediated effector functions, but not CD8+ T cells, for protection, making RABV infection an excellent mouse model to pinpoint the role for IL-21 in vaccine-induced immunity against RABV infections and potentially for other pathogens that rely solely on B cells for protection. In this report, our preliminary data indicate that IL-21 is critical for the development of effective vaccine-induced primary antibody responses against RABV infections by influencing GC B cells or PC generation in a vaccine dose-dependent manner, while also showing IL-21 is dispensable for RABV-specific secondary antibody responses when a primary antibody response develops. All animal work was reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Jefferson Medical College, Thomas Jefferson University. Work was completed in accordance with international standards [Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) ] and in compliance with Public Health Service Policy on Humane Care and Use of Laboratory Animals, The Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (NIH). The construction of the live RABV-based vaccine (rRABV) used in this study was described elsewhere and was previously named SPBN [9], [36], [37]. This vaccine is a molecular clone derived from the attenuated SAD-B19 vaccine strain of RABV. Virus stocks were propagated on baby hamster kidney cells and then concentrated and purified over a 20% sucrose cushion. The challenge virus used was the pathogenic Challenge Virus Strain-N2c (CVS-N2c), which is a mouse-adapted sublclone of CVS-21 RABV [38]. CVS-N2c was initially propagated in neonatal mouse brains and then passaged once in-vitro on a neuroblastoma cell line (NA cells). The titer of CVS-N2c required to kill unvaccinated mice was determined experimentally by inoculating serial ten-fold dilutions into naïve immune-competent mice [39] i. m. and then observing mice daily for clinical neurological symptoms of rabies. The titer required to kill unvaccinated mice within 8 days post-infection, which is typical for CVS-N2c [9], [10], was determined to be 105 focus forming units (ffu) /mouse. Cryopreserved embryos of mice deficient in the IL-21 receptor (B6; 129-IL21r tm1Wjl/Mmucd); #015505-UCD) [39] were obtained from the Mutant Mouse Regional Resource Centers (NIH) and implanted and bred in-house at Thomas Jefferson University in a pathogen-free animal facility. Control C57BL/6 mice were obtained from the Frederick National Laboratory for Cancer Research (NCI). The following antibodies where purchased from BD Biosciences, unless otherwise noted, and used for flow cytometry staining: APC-Cy7-B220 (clone RA3-6B2), PerCP-CY5. 5-CXCR5 (clone 2G8), APC-CD138 (clone 281-2), PE-Cy7-CD95/Fas (clone Jo2), FITC-GL7, eFluor 450-CD4 (clone RM4–5, eBioscience), PE-PD1 (clone J43, eBioscience), Alexa Fluor 700-CD38 (clone 90, eBioscience), rat anti-mouse CD16/32 (FcBlock; Pharmingen). Groups of 6- to 10-week-old female IL-21R−/− or wild-type C57BL/6 mice were inoculated intramuscularly (i. m.) with 103 or 105 focus forming units (ffu) /mouse with rRABV or an equivalent volume of PBS as controls. Five weeks post-immunization, mice were challenged i. m. with 105 ffu/mouse with CVS-N2c and then observed for three weeks for clinical signs of rabies. Mice were euthanized at the onset of neurological symptoms. At various times post-immunization and challenge, blood was collected retro-orbitally. Three-fold serial dilutions of sera were tested by ELISA to determine RABV G-specific IgG antibodies as described [10] and reported as the reciprocal serial dilution. Data represented two independent experiments (N = 9–11 mice per group). To measure virus neutralizing antibody titers, the Rapid Fluorescent Foci Inhibition Test (RFFIT) was completed on pooled sera from two independent experiments as described previously [16], [40], [41]. Groups of 6- to 10-week-old female IL-21R−/− or wild-type mice were inoculated i. m. with 103 or 105 ffu/mouse with rRABV or PBS/naive as controls. Draining lymph nodes and bone marrow cells were collected 7 and/or 14 days post-immunization. Single cell suspensions (106 cells/sample) were incubated with rat anti-mouse CD16/32 (1 ug/106 cells) in fluorescence-activated cell sorter (FACS) buffer (PBS supplemented with 2% fetal bovine serum) for 1 h on ice. Cells were washed twice with FACS buffer and incubated with fluorescently conjugated antibodies (0. 2 ug/106 cells) for 30 min at RT in the dark. Cells were subsequently washed 2 times with FACS buffer and fixed in 2% paraformaldehyde in PBS for 30 minutes. Flow cytometry was completed using FACScan (BD LSRII) and analyzed by FlowJo software. Data represents samples completed in duplicate (N = 3–5 mice) [16]. Kaplan-Meier survival curves were analyzed by the log rank test; *p = <0. 05 indicates significant survivorship between two immunization groups [9], [10]. Statistical difference between two groups of data was compared using an unpaired, two-tailed t test and data is presented at the mean ± SEM; *p<0. 05, **p = 0. 01–0. 001, ***p≤0. 001 [9], [10]. Cytokines present at the time of immunization have the ability to affect the outcome of vaccine-induced B cell responses. Due to the importance for IL-21 in promoting effective T-dependent B cell responses [21], [22], we examined the requirement for IL-21R signaling in the generation of antibodies in a mouse model of RABV immunogenicity and protection using a mouse model of IL-21R-deficiency. These mice, designated here as IL-21R−/− mice, lack the IL-21R extracellular and transmembrane domains but show normal lymphoid development [39]. T and B cells exhibit similar proliferative responses to CD3-speficic antibodies or LPS, respectively, when compared to wild-type controls [39]. As a group, IL-21R−/− mice immunized with a low dose of a live recombinant RABV-based vaccine (rRABV) (103 ffu/mouse) showed only low levels of anti-RABV G antibodies that were not significantly different from PBS-immunized mice at all time points tested post-immunization (Figure 1, left panels, a–e), although at least three IL-21R−/− mice developed anti-RABV antibodies by day 28 post-immunization (Figure 1B). The seroconversion of these three IL-21−/− mice may explain the limited protection observed in the pathogenic challenge experiments described later in this report. Wild-type mice immunized with the same dose of rRABV showed significant levels of anti-RABV G antibodies as early as 7 days post-inoculation compared to rRABV-immunized IL-21R−/− mice, and these antibody responses continued to increase through day 21 post-immunization. VNA titers detected 28 days post-immunization (Figure 1C) are consistent with the antibody titers detected by ELISA (Figure 1B) indicating anti-RV antibody titers detected by ELISA are representative of the ability for vaccine-induced antibodies to neutralize rabies virus. IL-21R−/− mice immunized with a higher dose of rRABV (105 ffu/mouse) showed significantly reduced anti-RABV antibody responses when compared to wild-type mice also immunized with 105 ffu/mouse of rRABV (Figure 1, right panels, f–j). Together, this data indicates that IL-21 is critical for the induction of optimal primary anti-RABV antibody responses, especially when low doses of vaccine are used. We next wanted to evaluate the effect of IL-21 on vaccine-induced antibody recall responses and protection against pathogenic RABV challenge. Five weeks post-immunization with rRABV, mice from Figure 1 were challenged with 105 ffu/mouse of a highly pathogenic mouse-adapted RABV strain (Challenge Virus Strain-N2c; CVS-N2c) [38], which typically kills naïve mice within 8 days post-infection [9], [10]. Consistent with the low antibody titers detected during the primary antibody response, significantly less anti-RABV antibodies were detected three or five days post-challenge in IL21R−/− mice immunized with 103 ffu/mouse of rRABV compared to the antibody recall response detected in wild-type mice (Figure 2, panels a and b, and Figure 2B) and antibody recall titers were not significantly different from PBS-immunized/CVS-N2c-challenged mice (Figure 2, e). Only 40% of IL-21R−/− mice immunized with 103 ffu/mouse of rRABV were protected against pathogenic RABV challenge, while all wild-type mice that were similarly immunized were protected against challenge (Figure 3, left panel). As expected, those mice with higher antibody titers showed protection compared to mice with lower antibody titers (Figure 2B). Conversely, an antibody recall response was induced in IL-21R−/− mice immunized with 105 ffu/mouse of rRABV within three days post-challenge with CVS-N2c at levels equivalent to rRABV-immunized wild-type mice (Figure 2, panels c and d) and all immunized IL-21R−/− mice were protected against pathogenic RABV challenge (Figure 3, right panel). Taken together, IL-21 is required for optimal primary (Figure 1) but not secondary (Figure 2) antibody responses to RABV vaccination. Furthermore, IL-21 is required for protection against pathogenic RABV in a vaccine dose-dependent manner (Figure 3). Next we wanted to determine whether the impaired primary anti-RABV antibody response in IL-21R−/− mice was due to an overall defect in the generation of Tfh and/or GC B cells. Lymph nodes from IL-21R−/− or wild-type mice were collected 7 or 14 days post-immunization with 103 or 105 ffu/mouse of rRABV or PBS alone to determine the influence of IL-21 on Tfh and B cell populations. Representative gating strategies [17], [42] to identify CD4+ T cells from the total live lymph node cultures (Figure 4A) or Tfh (CD4+CXCR5hiPD1hi) cells from the CD4+ T cell populations (Figure 4B) are shown. A significant increase in the number of CD4+ T cells displaying a Tfh phenotype was detected in IL-21R−/− mice 14 days post-immunization with 103 or 105 ffu/mouse rRABV compared to similarly immunized wild-type mice (Figure 4C and Figure 4D, respectively). However, the formation of optimal GC B cells appears to be dependent on the dose of vaccine administered (Figure 5). Representative gating strategies [22], [43], [44] to identify B220+ B cells from the total live lymph node cultures (Figure 5A) or GC B cells (B220+GL7hiCD95/Fashi) from the B220+ B cell population (Figure 5B) are shown. IL-21R−/− mice immunized with a low dose of vaccine failed to induce optimal GC B cell formation compared to wild-type mice 14 days post-immunization, as shown by a significant decrease in the number of GC B cells in IL-21R−/− mice compared to wild-type mice (Figure 5C). However, a significant increase in the number of GC B cells was detected in IL-21R−/− mice immunized with 105 ffu/mouse of rRABV compared to wild-type mice 14 days post-immunization (Figure 5D). The data indicates that the suboptimal primary antibody responses detected in IL-21R−/− mice immunized with 103 ffu/mouse of rRABV appears to be due to the lack of GC B cell formation, while the suboptimal primary antibody response detected in IL-21R−/− mice immunized with 105 ffu/mouse most likely does not result from a defect in Tfh or GC B cell development. Our analysis above shows that IL-21R signaling is dispensable for the formation of the Tfh and GC B cell populations in response to higher doses of rRABV, indicating that other B cell types are more likely responsible for the suboptimal primary antibody titers detected in IL-21R−/− mice immunized with 105 ffu/mouse. Since IL-21 can also influence the balance between the generation of memory B cells and PCs [24], [45]–[47], we investigated the role for IL-21R signaling to regulate memory B cell and PCs populations in response to RABV vaccination. Figure 6A and Figure 6B show representative gating strategies [48]–[50] to identify the memory B220+ B cells (CD38+CD138−) from the lymph node and PC (B220loCD138+) populations from the bone marrow from mice immunized with 103 or 105 ffu/mouse of rRABV or PBS alone. The presence or absence of IL-21R does not appear to influence the development of memory B cells in mice immunized with 103 ffu/mouse of rRABV (Figure 6C). However, the percentage of memory B cells was significantly increased in the lymph node cell cultures from IL-21R−/− mice immunized with 105 ffu/mouse of rRABV as early as 7 days post-immunization compared to immunized wild-type mice (Figure 6C). By day 14 post-immunization, similar memory B cell populations were measured (data not shown), indicating that IL-21 is not required for the formation of memory B cells in response to live RABV-based vaccination. This is consistent with the findings in Figure 1 and Figure 2 indicating that IL-21 is dispensable for secondary antibody responses against RABV infection when a primary antibody response develops. However, the percentage of PCs was reduced in the bone marrow of IL-21R−/− mice immunized with 105 ffu/mouse compared to rRABV- or PBS-immunized wild-type mice 14 days post-immunization (Figure 6D), consistent with the suboptimal primary antibody titers detected in IL-21R−/− mice. Current rabies PEP regimens are based on multiple doses of inactivated RABV-based vaccines administered intramuscularly or intradermally. In cases of severe exposure, rabies immune globulin (RIG) is administered [51]–[53]. The development of a single-dose vaccine would greatly benefit human rabies prevention by reducing the cost of vaccination and saving lives. Understanding immune parameters that influence the magnitude and/or quality of anti-RABV antibody responses may lead to more effective single-dose vaccines [54]. Correlates of protection against rabies infections are defined as virus neutralizing antibodies directed against the single viral transmembrane glycoprotein (G) [15], [52], [54], [55]. CD8+ T cells do not appear to be important for the clearance of RABV infections [56]. Protection against RABV infection typically requires CD4+ T cell help [56]–[60], although we recently showed that this requirement is not absolute and that protection against pathogenic RABV challenge can be afforded in mice devoid of all T cells (TCRβδ−/− mice) vaccinated with a matrix gene-deleted RABV-based vaccine (rRABV-ΔM) [16]. Furthermore, we show that mice immunized with rRABV-ΔM also induce antibodies by T cell-dependent extrafollicular B cell responses before GC-derived B cells are detected. Together, our previous work identified multiple pathways of B cell development that can be exploited to make more efficacious RABV-based vaccines for use in humans. Nonetheless, very little information is available on how effective B cells develop in response to live RABV-based vaccination. IL-21 is a pleotropic cytokine that is produced by NKT cells and CD4+ T cells, most notably Th17 and Tfh cells. IL-21 binds to the IL-21R on a wide variety of cells involved in innate immunity, including DCs, NK cells, NKT cells, and macrophages, as well as on cells involved in adaptive immunity, such as B cells and CD4+ or CD8+ T cells [reviewed in [20]]. Due to its multiple roles in innate and adaptive immunity, IL-21 has the potential to influence the quality and magnitude of vaccine-induced immunity to acute viral infections. Here we used a mouse model of IL-21R-deficiency to evaluate the role for IL-21R signaling in vaccine-induced protection against RABV; i. e., an acute viral infection that relies on B cells for protection that has implications for global public health initiatives. In this report, we showed that IL-21R signaling is critical for the generation of optimal primary anti-RABV antibody responses to vaccination. Primary anti-RABV antibody titers were significantly reduced in immunized IL-21R−/− mice compared to wild-type mice at almost all time points tested post-immunization, suggesting IL-21R signaling plays important roles throughout RABV-specific primary B cell responses. Nonetheless, IL-21R signaling appears to influence immunity in a vaccine dose-dependent manner, which is consistent with findings by others suggesting the influence of IL-21 is dependent on the model studied [22], [61]. Indeed, significantly less IL-21R−/− mice immunized with low-dose vaccination were protected against pathogenic challenge compared to wild-type mice while all IL-21R−/− and wild-type mice immunized with high-doses of vaccine survived challenge similarly. Despite the differences in protection elicited in IL-21R−/− mice immunized with different doses of vaccine, it appears that IL-21 is critical for the generation of optimal RABV-specific primary B cell responses. One potential explanation for the suboptimal primary antibody responses observed in immunized IL-21R−/− mice compared to immunized wild-type mice might be that GC B cells failed to form in IL-21R−/− mice, therefore, the GC B cell compartment was analyzed in mice immunized with different doses of vaccine. GC-derived B cells were reduced in IL-21R−/− mice immunized with a low dose of vaccine compared to similarly immunized wild-type mice, indicating that IL-21 is required for optimal GC B cell formation in response to low-dose RABV-based vaccination. On the other hand, GC-derived B cells expanded in IL-21R−/− mice immunized with a high dose of vaccine compared to similarly immunized wild-type mice, indicating that IL-21 is dispensable for GC B cell formation after high-dose vaccination with rRABV-based vaccines. Furthermore, the data indicates that factors other than IL-21 were responsible for GC B cell formation in IL21R−/− mice immunized with higher doses of vaccine. Multiple signals lead to B cell activation and functions. These signals can come from BCR or Toll-like receptor (TLR) ligation, TNF superfamily receptor engagement (eg., via BAFF and APRIL) or cytokine signaling. Furthermore, B cell activation is contextual, meaning B cells are differentially activated in the presence of different signals at the time of antigen exposure. Due to the repetitive display of rabies antigen on the surface of infectious particles, the potential exists that cross-linking BCRs and/or TLRs on the surface of B cells overcame the requirement for IL-21R signaling in B cell activation when high doses of vaccine are administered. The influences of these and other B cell signaling events in the context of RABV-based vaccine-induced B cell activation were not directly measured in these studies and remain to be elucidated. Nonetheless, based on the results reported here, it appears that IL-21R signaling is important for optimal primary vaccine-induced antibody responses to RABV vaccination especially when low doses of vaccine are administered. As noted above, a rapid antibody response is critical for rabies PEP to neutralize virus before it reaches the CNS. We have recently shown that RABV-based vaccines are able to induce early and rapid T cell-dependent extrafollicular antibody responses before GC B cells are formed [16]. These early pre-GC B cell responses contributed to the protection against pathogenic RV challenge early post-immunization, which is an important attribute for PEP [16]. In the studies described in this report, we detected a significant reduction in antibody titers in IL-21R−/− mice as early as 5 days post-immunization with a high dose of rRABV compared to immunized wild-type mice, suggesting that IL-21 may be influencing the outcome of extrafollicular antibody responses in the context of RABV vaccination, although this was not directly studied in this report. Nonetheless, the frequency of Tfh and GC B cells was similar in IL-21R−/− mice compared to wild-type mice 7 days post-immunization and, therefore, it would appear that the early suboptimal antibody responses in IL-21R−/− mice may be due to impaired extrafollicular PCs directly and not through impaired Tfh or GC B cell formation. This is consistent with the findings that IL-21 can promote Blimp-1 expression and PC development [47], IL-21- or IL-21R-deficiency decreases extrafollicular PCs in a model of NP-KLH immunity [62], and that IL-21 acts on early stages of B cell differentiation before GC or PC B cells are formed [42]. Finally, IL-21 has been reported to be important for Tfh cell maintenance but not formation. Together, existing data suggests that IL-21R−/− signaling influences early events in pre-GC B cell development in the context of RABV vaccination [22]. IL-21 can also influence the balance of B cell differentiation into memory B cells or PCs [45]–[47]. The specific role for IL-21 in memory B cell responses is not completely clear and appears to rely on the type of antigen used and the model studied [61], [62]. In the context of RABV vaccination, IL-21R signaling was not required for the generation of B cells displaying a memory B cell phenotype in IL-21R−/− mice immunized with either vaccine dose. This is consistent with our finding showing that IL-21R signaling is not required for optimal secondary anti-RABV G antibody titers after challenge with pathogenic RABV. However, we detected a decrease in the number of PCs in IL-21R−/− mice immunized with either dose of vaccine compared to wild-type mice. We cannot determine whether the slightly suboptimal PC subset detected in IL-21R−/− mice immunized with low doses of vaccine was indirectly a result of impaired GC B cell development or directly as a result of impaired PC formation itself. However, in mice immunized with a high dose of vaccine where we observed an expansion of GC-derived B cells in IL-21R−/− mice, we also observed a decrease in PCs in the bone marrow compared to wild-type mice, indicating that IL-21 acts directly on the formation of PCs. Together, the data shows that IL-21 influences the balance between memory and PC B cell formation in the context of RABV vaccination. Despite the impaired primary antibody response and PC B cell formation in immunized IL-21R−/− mice, we detected an expansion of CD4+ T cells displaying a pre-Tfh (data not shown), Tfh cell and GC B cell phenotype in IL-21R−/− mice compared to wild-type mice at 14 days post-immunization. The expansion of GC B cells and Tfh cells in the absence of IL-21R signaling was also shown by King I. L. et al in a model of Heligmosomoides polygyrus immunity [24], suggesting that IL-21R signaling may not be necessary for the generation of these cell types in response to a wide range of pathogens or vaccination. Furthermore, the increase in GC B cells and Tfh cells in H. polygyrus-infected or RABV-vaccinated IL-21R−/− mice suggests that IL-21R signaling may play an inhibitory role in the development of T and B cells in the context of some pathogens, which is consistent with the ability for IL-21 to activate or inhibit immune function depending on the antigen and available co-stimulatory signals [20]. The expansion of Tfh and GC B cells in IL-21R−/− mice compared to wild type mice is also consistent with the finding that IL-21 has the ability to mediate apoptosis in primary resting and activated murine B or to promote apoptosis or growth arrest for non-specifically activated B cells [63]. Alternatively, the elevated number of GC Tfh cells could be a result of the lack of PC that developed in the IL21R−/− mice. Pelletier et al described a negative regulatory feedback-loop in which antigen-specific PCs negatively regulate antigen-specific Tfh cell development and function [64]. In this report, they also observed a significant expansion of Tfh cells and GC B cells in the absence of PC development. Together, the role for IL-21 in the homeostatic balance of T and B cell development in the context of infectious diseases appears to be important and remains to be fully elucidated. Additional studies are needed to identify the exact cell type (s) responsible for the affects described in this report. While we speculate that B cell-intrinsic IL-21R signaling is responsible for the induction of optimal anti-RABV antibody responses, we cannot rule out the influence of other cell types that also express IL-21R. IL-21 has the ability to influence the function of macrophages, NK cells and NKT cells by affecting survival/apoptosis, antigen processing, and cytokine secretion [reviewed in [20]]. The function of these cells of the innate immune system may indirectly be affecting the outcome of B or T cell functions in the context of RV vaccination. Nonetheless, IL-21 has the potential to influence a wide range of B cell functions and pathways. Our preliminary data indicates that IL-21 is critical for the formation of optimal vaccine-induced primary antibody responses and demonstrates an important role for IL-21 in the generation of vaccine-induced immunity against RABV infection and perhaps other acute infections that rely on B cell-mediated effector functions for protection.

Title: Investigating the Role for IL-21 in Rabies Virus Vaccine-induced Immunity Summary: Over two-thirds of the world' s population lives in regions where rabies is endemic, resulting in over 15 million people receiving post-exposure treatment. A person, disproportionately a child, dies of rabies every 20 minutes and the cost of rabies prevention exceeds $1 billion US dollars per year. The development of a single-dose human rabies vaccine would greatly reduce the burden of rabies globally by lowering the cost associated with rabies vaccination and saving lives. Understanding how B cells develop to produce protective virus neutralizing antibodies would greatly help to achieve the goal of developing a single-dose vaccine. In this report, we show that IL-21 is critical for the induction of primary vaccine-induced anti-RABV G antibody titers and that the effects of IL-21 are highly dependent on the dose of vaccine administered. In our model of rabies immunogenicity and protection, the lack of IL-21 receptor influenced the detection of B cells in germinal centers in lymph nodes or of plasma cells in bone marrow after immunization with low or high doses of vaccine, respectively. Overall, these preliminary results indicate that IL-21 has the potential to influence B cell development and functions in the context of rabies vaccine-induced immunity and protection.

lay_plos

en

Write a title and summarize: SECTION 1. SHORT TITLE. This Act may be cited as the ``Marine Mineral Resources Research Act of 1996''. SEC. 2. RESEARCH PROGRAM. The Mining and Minerals Policy Act of 1970 (30 U.S.C. 21a) is amended-- (1) by inserting after the first section the following: ``TITLE I--MINING POLICY''; (2) by redesignating section 2 as section 101; and (3) by adding at the end the following: ``TITLE II--MARINE MINERAL RESOURCES RESEARCH PROGRAM ``SEC. 201. DEFINITIONS. ``In this title: ``(1) The term `contract' has the same meaning as `procurement contract' in section 6303 of title 31, United States Code. ``(2) The term `cooperative agreement' has the same meaning as in section 6305 of title 31, United States Code. ``(3) The term `eligible entity' means-- ``(A) a research or educational entity chartered or incorporated under Federal or State law; ``(B) an individual who is a United States citizen; or ``(C) a State or regional agency. ``(4) The term `grant' has the same meaning as `grant agreement' in section 6304 of title 31, United States Code. ``(5) The term `in-kind contribution' means a noncash contribution provided by a non-Federal entity that directly benefits and is related to a specific project or program. An in- kind contribution may include real property, equipment, supplies, other expendable property, goods, and services. ``(6) The term `marine mineral resource' means-- ``(A) sand and aggregates; ``(B) placers; ``(C) phosphates; ``(D) manganese nodules; ``(E) cobalt crusts; ``(F) metal sulfides; and ``(G) other marine resources that are not-- ``(i) oil and gas; ``(ii) fisheries; or ``(iii) marine mammals. ``(7) The term `Secretary' means the Secretary of the Interior. ``SEC. 202. RESEARCH PROGRAM. ``(a) In General.--The Secretary shall establish and carry out a program of research on marine mineral resources. ``(b) Program Goal.--The goal of the program shall be to-- ``(1) promote research, identification, assessment, and exploration of marine mineral resources in an environmentally responsible manner; ``(2) assist in developing domestic technologies required for efficient and environmentally sound development of marine mineral resources; ``(3) coordinate and promote the use of technologies developed with Federal assistance, and the use of available Federal assets, for research, identification, assessment, exploration, and development of marine mineral resources; and ``(4) encourage academia and industry to conduct basic and applied research, on a joint basis, through grants, cooperative agreements, or contracts with the Federal Government. ``(c) Responsibilities of the Secretary.--In carrying out the program, the Secretary shall-- ``(1) promote and coordinate partnerships between industry, government, and academia to research, identify, assess, and explore marine mineral resources in an environmentally sound manner; ``(2) undertake programs to develop the basic information necessary to the long-term national interest in marine mineral resources (including seabed mapping) and to ensure that data and information are accessible and widely disseminated as needed and appropriate; ``(3) identify, and promote cooperation among agency programs that are developing, technologies developed by other Federal programs that may hold promise for facilitating undersea applications related to marine mineral resources, including technologies related to vessels and other platforms, underwater vehicles, survey and mapping systems, remote power sources, data collection and transmission systems, and various seabed research systems; and ``(4) foster communication and coordination between Federal and State agencies, universities, and private entities concerning marine mineral research on seabeds of the continental shelf, ocean basins, and arctic and cold water areas. In carrying out these responsibilities, the Secretary shall ensure the participation of non-Federal users of technologies and data related to marine mineral resources in planning and priority setting. ``SEC. 203. GRANTS, CONTRACTS, AND COOPERATIVE AGREEMENTS. ``(a) Assistance and Coordination.-- ``(1) In general.--The Secretary shall award grants or contracts to, or enter into cooperative agreements with, eligible entities to support research for the development or utilization of-- ``(A) methods, equipment, systems, and components necessary for the identification, assessment, and exploration of marine mineral resources in an environmentally responsible manner; ``(B) methods of detecting, monitoring, and predicting the presence of adverse environmental effects in the marine environment and remediating the environmental effects of marine mineral resource exploration, development, and production; and ``(C) education and training material in marine mineral research and resource management. ``(2) Cost-sharing for contracts or cooperative agreements.-- ``(A) Federal share.--Except as provided in subparagraph (B)(ii), the Federal share of the cost of a contract or cooperative agreement carried out under this subsection shall not be greater than 80 percent of the total cost of the project. ``(B) Non-federal share.--The remaining non-Federal share of the cost of a project carried out under this section may be-- ``(i) in the form of cash or in-kind contributions, or both; and ``(ii) comprised of funds made available under other Federal programs, except that non-Federal funds shall be used to defray at least 10 percent of the total cost of the project. ``(C) Consultation.--Not later than 180 days after the date of enactment of this Act, the Secretary shall establish, after consultation with other Federal agencies, terms and conditions under which Federal funding will be provided under this subsection that are consistent with the Agreement on Subsidies and Countervailing Measures referred to in section 101(d)(12) of the Uruguay Round Agreement Act (19 U.S.C. 3511(d)(12)). ``(b) Competitive Review.-- ``(1) In general.--An entity shall not be eligible to receive a grant or contract, or participate in a cooperative agreement, under subsection (a) unless-- ``(A) the entity submits a proposal to the Secretary at such time, in such manner, and accompanied by such information as the Secretary may reasonably require; and ``(B) the proposal has been evaluated by a competitive review panel under paragraph (3). ``(2) Competitive review panels.-- ``(A) Composition.--A competitive review panel shall be chaired by the Secretary or by the Secretary's designee and shall be composed of members who meet the following criteria: ``(i) Appointment.--The members shall be appointed by the Secretary. ``(ii) Experience.--Not less than 50 percent of the members shall represent or be employed by private marine resource companies that are involved in exploration of the marine environment or development of marine mineral resources. ``(iii) Interest.--None of the members may have an interest in a grant, contract, or cooperative agreement being evaluated by the panel. ``(B) No compensation.--A review panel member who is not otherwise a Federal employee shall receive no compensation for performing duties under this section, except that, while engaged in the performance of duties away from the home or regular place of business of the member, the member may be allowed travel expenses, including per diem in lieu of subsistence, in the same manner as a person employed intermittently in the Government service under section 5703 of title 5, United States Code. ``(3) Evaluation.--A competitive review panel shall base an evaluation of a proposal on criteria developed by the Secretary that shall include-- ``(A) the merits of the proposal; ``(B) the research methodology and costs of the proposal; ``(C) the capability of the entity submitting the proposal and any other participating entity to perform the proposed work and provide in-kind contributions; ``(D) the amount of matching funds provided by the entity submitting the proposal or provided by other Federal, State, or private entities; ``(E) the extent of collaboration with other Federal, State, or private entities; ``(F) in the case of a noncommercial entity, the existence of a cooperative agreement with a commercial entity that provides for collaboration in the proposed research; ``(G) whether the proposal promotes responsible environmental stewardship; and ``(H) such other factors as the Secretary considers appropriate. ``(c) Limitations.-- ``(1) Administrative expenses.--Not more than 10 percent of the amount made available to carry out this section during a fiscal year may be used by the Secretary for expenses associated with administration of the program authorized by this section. ``(2) Construction costs.--None of the funds made available under this section may be used for the construction of a new building or the acquisition, expansion, remodeling, or alteration of an existing building (including site grading and improvement and architect fees). ``(d) Reports.--An eligible entity that receives a grant or contract or enters into a cooperative agreement under this section shall submit an annual progress report and a final technical report to the Secretary that-- ``(1) describes project activities, implications of the project, the significance of the project to marine mineral research, identification, assessment, and exploration, and potential commercial and economic benefits and effects of the project; and ``(2) in the case of an annual progress report, includes a project plan for the subsequent year. ``SEC. 204. MARINE MINERAL RESEARCH CENTERS. ``(a) In general.--No later than 90 days after the date of enactment of this section, the Secretary shall designate 3 centers for marine mineral research and related activities. ``(b) Concentration.--One center shall concentrate primarily on research in the continental shelf regions of the United States, 1 center shall concentrate primarily on research in deep seabed and near- shore environments of islands, and 1 center shall concentrate primarily on research in arctic and cold water regions. ``(c) Criteria.--In designating a center under this section, the Secretary shall give priority to a university that-- ``(1) administers a federally funded center for marine minerals research; ``(2) matriculates students for advanced degrees in marine geological sciences, nonenergy natural resources, and related fields of science and engineering; ``(3) is a United States university with established programs and facilities that primarily focus on marine mineral resources; ``(4) has engaged in collaboration and cooperation with industry, governmental agencies, and other universities in the field of marine mineral resources; ``(5) has demonstrated significant engineering, development, and design experience in two or more of the following areas; ``(A) seabed exploration systems; ``(B) marine mining systems; and ``(C) marine mineral processing systems; and ``(6) has been designated by the Secretary as a State Mining and Mineral Resources Research Institute. ``(d) Center Activities.--A center shall-- ``(1) provide technical assistance to the Secretary concerning marine mineral resources; ``(2) advise the Secretary on pertinent international activities in marine mineral resources development; ``(3) engage in research, training, and education transfer associated with the characterization and utilization of marine mineral resources; and ``(4) promote the efficient identification, assessment, exploration, and management of marine mineral resources in an environmentally sound manner. ``(e) Allocation of Funds.--In distributing funds to the centers designated under subsection (a), the Secretary shall, to the extent practicable, allocate an equal amount to each center. ``(f) Limitations.-- ``(1) Administrative expenses.--Not more than 5 percent of the amount made available to carry out this section during a fiscal year may be used by the Secretary for expenses associated with administration of the program authorized by this section. ``(2) Construction costs.--None of the funds made available under this section may be used for the construction of a new building or the acquisition, expansion, remodeling, or alteration of an existing building (including site grading and improvement and architect fees). ``SEC. 205. AUTHORIZATION OF APPROPRIATIONS. ``There is authorized to be appropriated such sums as are necessary to carry out this title.''. Speaker of the House of Representatives. Vice President of the United States and President of the Senate.

Title: Marine Mineral Resources Research Act of 1996 Summary: Marine Mineral Resources Research Act of 1996 - Amends the Mining and Minerals Policy Act of 1970 to direct the Secretary of the Interior to: (1) establish and implement a marine mineral resources research program; (2) award grants and contracts and enter into cooperative agreements for the attendant research and development undertaken by industry, government, and academic sectors; and (3) establish terms and conditions under which Federal funding will be provided consistent with the Agreement on Subsidies and Countervailing Measures referred to in the Uruguay Round Agreement Act. Prescribes guidelines for competitive review of such Federal grants, contracts, and cooperative agreements. Directs the Secretary to designate three marine mineral research centers, one in the continental shelf regions of the United States, the second in deep seabed and near-shore environments of islands, and the third in arctic and cold water regions. Restricts administrative expenses to not more than five percent during a fiscal year. Authorizes appropriations.

billsum

en

Summarize: By. Peter Rook For The Mail On Sunday. Convicted sex offenders could pick up passengers in licensed minicabs under new Government plans, according to critics who say they pose a ‘real danger’ to the British public. The Government’s Deregulation Bill, which will be debated in the Lords next month, could permit anyone to drive a licensed vehicle without going through a council vetting process. The proposals come as the full extent of taxi drivers’ involvement in the Rotherham child abuse scandal – in which some victims were raped in cabs after being picked up outside schools – continues to emerge. The Government’s Deregulation Bill, which will be debated in the Lords next month, could permit anyone to drive a licensed vehicle without going through a council vetting process. Currently, anyone looking to operate as a private hire vehicle operator must obtain a licence from a council, which checks for previous criminal convictions and can refuse licences if it feels the applicant is not a ‘fit and proper person’. Operators are able to operate only in the area in which they are licensed. Under deregulation, a licensed driver could operate outside his area and enforcement officers in that area would not be able to check if he has been vetted. It will also allow licensed private hire operators to pass work to drivers who may not have been through the vetting process. Critics have hit out at the move. Rachel Griffin, director of personal safety campaign group the Suzy Lamplugh Trust, said: ‘The changes are going to put people across the country in real danger as they will make it much easier for someone with a past of violence and sexual offences to pose as a legitimate driver.’ A Department of Transport spokesman said: ‘The Deregulation Bill will not put taxi passengers at risk and drivers will continue to have their backgrounds routinely checked. Councils will have strong tools to assess drivers’ and operators’ suitability and to carry out enforcement activity. ‘The Disclosure and Barring Service [which replaced the criminal Records Bureau] will allow licensing authorities to discover any new convictions during the lifetime of a driver’s licence.’ Police estimate there are more than 1,000 sexual assaults each year involving unlicensed cab drivers in London alone

Summary: Government's Deregulation Bill will be debated in the Lords next month. Would allow anyone to drive licensed vehicle without vetting process. Critics say new law would put passengers 'across country in real danger' Proposals follow revelation some Rotherham victims were raped in cabs. Last week The Mail on Sunday reported there are more than 1,000 sexual assaults each year involving taxi drivers in London. This police figure relates solely to unlicensed cab drivers. The Mail on Sunday apologises for the error.

cnn_dailymail

en

Write a title and summarize: SECTION 1. SHORT TITLE; TABLE OF CONTENTS. (a) Short Title.--This Act may be cited as the ``Small Business Health Relief Act of 2015''. (b) Table of Contents.--The table of contents for this Act is as follows: Sec. 1. Short title; table of contents. TITLE I--MAKING COVERAGE AFFORDABLE FOR SMALL BUSINESSES Sec. 101. Protecting American jobs and wages. Sec. 102. Increasing flexibility for small businesses. Sec. 103. Increasing choices for Americans. Sec. 104. Protecting patients from higher premiums. Sec. 105. Ensuring affordable coverage. TITLE II--INCREASING CONSUMER CONTROL Sec. 201. Repeal of restriction on over-the-counter medicines. Sec. 202. Repeal of the annual cap. TITLE III--ALLOWING INDIVIDUALS TO KEEP COVERAGE THEY LIKE Sec. 301. Allowing individuals to keep the coverage they have if they like it. TITLE I--MAKING COVERAGE AFFORDABLE FOR SMALL BUSINESSES SEC. 101. PROTECTING AMERICAN JOBS AND WAGES. (a) Repeal of Shared Responsibility Payment for Employers Regarding Health Coverage.-- (1) In general.--Chapter 43 of the Internal Revenue Code of 1986 is amended by striking section 4980H. (2) Conforming amendments.-- (A) The table of sections for chapter 43 of the Internal Revenue Code of 1986 is amended by striking the item relating to section 4980H. (B) Section 1311(d)(4)(I) of the Patient Protection and Affordable Care Act is amended by inserting ``and'' at the end of clause (i) and by striking clause (ii). (C) Section 1332(a)(2)(D) of such Act is amended by striking ``36B, 4980H, and 5000A'' and inserting ``36B and 5000A''. (D) Section 1411(e)(4)(B) of such Act is amended by striking clause (iii). (E) Section 1411(f) of such Act is amended to read as follows: ``(f) Appeals and Redeterminations.--The Secretary, in consultation with the Secretary of the Treasury, the Secretary of Homeland Security, and the Commissioner of Social Security, shall establish procedures by which the Secretary or one of such other Federal officers-- ``(1) hears and makes decisions with respect to appeals of any determination under subsection (e); and ``(2) redetermines eligibility on a periodic basis in appropriate circumstances.''. (F) Section 1411 of such Act is amended by striking subsection (i). (G) Section 1412(a)(2) of such Act is amended to read as follows: ``(2) the Secretary notifies the Exchange and the Secretary of the Treasury of the advance determinations; and''. (H) Section 1513 of such Act is amended by striking subsection (c). (3) Effective date.--The amendments made by this subsection shall apply to months after December 31, 2013. (b) Repeal of Reporting of Employer Health Insurance Coverage.-- (1) In general.--Subpart D of part III of subchapter A of chapter 61 of the Internal Revenue Code of 1986 is amended by striking section 6056. (2) Conforming amendments.-- (A) Section 6724(d)(1)(B) of the Internal Revenue Code of 1986 is amended by inserting ``or'' at the end of clause (xxiii), by striking ``, or'' at the end of clause (xxiv) and inserting a period, and by striking clause (xxv). (B) Section 6724(d)(2) of such Code is amended by inserting ``or'' at the end of subparagraph (FF), by striking ``, or'' at the end of subparagraph (GG) and inserting a period, and by striking subparagraph (HH). (3) Effective date.--The amendments made by this subsection shall apply to periods beginning after December 31, 2013. SEC. 102. INCREASING FLEXIBILITY FOR SMALL BUSINESSES. Section 1302(c)(2) of the Patient Protection and Affordable Care Act (Public Law 111-148) is repealed. SEC. 103. INCREASING CHOICES FOR AMERICANS. (a) Qualified Health Plan Coverage Satisfied by High Deductible Health Plan With Health Savings Account.--Section 1302(e) of the Patient Protection and Affordable Care Act (42 U.S.C. 18022(e)) is amended to read as follows: ``(e) High Deductible Health Plan With Health Savings Account.--A health plan not providing a bronze, silver, gold, or platinum level of coverage shall be treated as meeting the requirements of subsection (d) with respect to any plan year for any enrollee if the plan meets the requirements for a high deductible health plan under section 223(c)(2) of the Internal Revenue Code of 1986 and such enrollee has established a health savings account (as defined in section 223(d)(1) of such Code) in relation to such plan.''. (b) Conforming Amendments.-- (1) Subparagraph (C) of section 1312(d)(3) of the Patient Protection and Affordable Care Act (42 U.S.C. 18032(d)(3)) is amended by striking ``, except'' and all that follows through ``1302(e)(2)''. (2) Subparagraph (A) of section 36B(c)(3) of the Internal Revenue Code of 1986, as added by section 1401(a) of the Patient Protection and Affordable Care Act (Public Law 111-148) is amended by striking ``, except'' and all that follows through ``such Act''. (3) Subparagraph (B) of section 1334(c)(1) of the Patient Protection and Affordable Care Act (42 U.S.C. 18054(c)(1)) is amended by striking ``and catastrophic coverage''. SEC. 104. PROTECTING PATIENTS FROM HIGHER PREMIUMS. Section 9010 of the Patient Protection and Affordable Care Act (Public Law 111-148), as amended by section 10905 of such Act, is repealed. SEC. 105. ENSURING AFFORDABLE COVERAGE. Section 2701(a)(1)(A)(iii) of the Public Health Service Act (42 U.S.C. 300(a)(1)(A)(iii)), as added by section 1201 of the Patient Protection and Affordable Care Act (Public Law 111-148), is amended by striking ``, except'' and all that follows through ``2707(c))''. TITLE II--INCREASING CONSUMER CONTROL SEC. 201. REPEAL OF RESTRICTION ON OVER-THE-COUNTER MEDICINES. (a) HSAs.--Section 223(d)(2)(A) of the Internal Revenue Code of 1986 is amended by striking the last sentence thereof. (b) Archer MSAs.--Section 220(d)(2)(A) of the Internal Revenue Code of 1986 is amended by striking the last sentence thereof. (c) Health Flexible Spending Arrangements and Health Reimbursement Arrangements.--Section 106 of the Internal Revenue Code of 1986 is amended by striking subsection (f). (d) Effective Date.-- (1) Distributions from savings accounts.--The amendments made by subsections (a) and (b) shall apply to amounts paid with respect to taxable years beginning after December 31, 2014. (2) Reimbursements.--The amendment made by subsection (c) shall apply to expenses incurred with respect to taxable years beginning after December 31, 2014. SEC. 202. REPEAL OF THE ANNUAL CAP. (a) In General.--Section 125 of the Internal Revenue Code of 1986 is amended by striking subsection (i) and by redesignating subsections (j) and (k) as subsections (i) and (j), respectively. (b) Effective Date.--The amendments made by this section shall apply to taxable years beginning after December 31, 2014. TITLE III--ALLOWING INDIVIDUALS TO KEEP COVERAGE THEY LIKE SEC. 301. ALLOWING INDIVIDUALS TO KEEP THE COVERAGE THEY HAVE IF THEY LIKE IT. (a) In General.--Section 1251(a)(2) of the Patient Protection and Affordable Care Act (42 U.S.C. 18011) is amended-- (1) by striking ``Except as provided in paragraph (3),'' and inserting the following: ``(A) In general.--Except as provided in paragraphs (3) and (4),''; and (2) by adding at the end the following: ``(B) Protecting employers and consumers with grandfathered coverage.-- ``(i) In general.--A group health plan or health insurance coverage in which an individual is enrolled on or after March 23, 2010, but before any plan year beginning not later than 1 year after the date of the enactment of this subparagraph, and which is deemed to be a grandfathered health plan under this section, shall continue to be considered a grandfathered health plan with respect to such individual regardless of any modification to the cost-sharing levels, employer contribution rates, or covered benefits under such plan or coverage as otherwise permitted under this Act (and the amendments made by this Act). ``(ii) Regulations.--The Secretary shall promulgate regulations to clarify the application of clause (i) to a plan or coverage that continues to be a grandfathered health plan pursuant to such clause.''. (b) Effective Date; Previously Promulgated Regulations Voided.-- (1) Effective date.--The amendments made by this section shall take effect as if included in the enactment of the Patient Protection and Affordable Care Act. (2) Previously promulgated regulations voided.--Any regulations relating to section 1251(a)(2) of such Act promulgated before the date of the enactment of this Act shall have no force or effect.

Title: Small Business Health Relief Act of 2015 Summary: Small Business Health Relief Act of 2015 Repeals provisions of the Internal Revenue Code that: (1) impose fines on large employers (those with 50 or more full-time employees) who fail to offer their full-time employees the opportunity to enroll in minimum essential health insurance coverage, and (2) require large employers to file a report with the Department of the Treasury on health insurance coverage provided to their full-time employees. Repeals provisions of the Patient Protection and Affordable Care Act (PPACA) that: (1) limit the annual deductible on health plans offered in the small group market, (2) deem catastrophic plans to meet essential health benefits coverage requirements for certain individuals, and (3) impose an annual fee on health insurance entities. Deems high deductible health plans to meet essential health benefits coverage requirements if the enrollee has established a health savings account. Amends the Public Health Service Act to repeal the limitation on premium rate variance by age in the individual or small group market. Repeals the prohibitions on payments for over-the-counter medications from health savings accounts, medical savings accounts, and health flexible spending arrangements. Repeals the $2,500 annual limit on employee contributions by salary reduction to a health flexible spending arrangement under a cafeteria plan. Allows a health plan to maintain its status as a grandfathered health plan regardless of any modification to cost-sharing, employer contribution rates, or covered benefits. Makes this allowance effective as if included in PPACA.

billsum

en

Write a title and summarize: Clonally derived bacterial populations exhibit significant genotypic and phenotypic diversity that contribute to fitness in rapidly changing environments. Here, we show that serial passage of Salmonella enterica serovar Typhimurium LT2 (StLT2) in broth, or within a mouse host, results in selection of an evolved population that inhibits the growth of ancestral cells by direct contact. Cells within each evolved population gain the ability to express and deploy a cryptic “orphan” toxin encoded within the rearrangement hotspot (rhs) locus. The Rhs orphan toxin is encoded by a gene fragment located downstream of the “main” rhs gene in the ancestral strain StLT2. The Rhs orphan coding sequence is linked to an immunity gene, which encodes an immunity protein that specifically blocks Rhs orphan toxin activity. Expression of the Rhs orphan immunity protein protects ancestral cells from the evolved lineages, indicating that orphan toxin activity is responsible for the observed growth inhibition. Because the Rhs orphan toxin is encoded by a fragmented reading frame, it lacks translation initiation and protein export signals. We provide evidence that evolved cells undergo recombination between the main rhs gene and the rhs orphan toxin gene fragment, yielding a fusion that enables expression and delivery of the orphan toxin. In this manner, rhs locus rearrangement provides a selective advantage to a subpopulation of cells. These observations suggest that rhs genes play important roles in intra-species competition and bacterial evolution. Bacteria often reside in complex communities such as biofilms in which cells from multiple species touch one another in a three-dimensional network [1]. These environments provide opportunities for cellular interactions, yet the mechanisms underlying contact-dependent competition and cooperation have been largely unexplored until recently. A diverse family of YD-peptide repeat proteins mediates at least two distinct forms of contact-dependent competition in Gram-negative and -positive bacteria [2]. The Rhs (rearrangement hotspot) proteins of Gram-negative enterobacteria [3], [4] are large (∼1,400–1,700 residues) toxic effectors that appear to be exported through the type VI secretion machinery. Related WapA (wall-associated protein A) proteins from Gram-positive bacteria are somewhat larger (∼2,200–3,600 residues) [5] and are likely exported through the general secretory pathway [2]. Rhs and WapA proteins are both characterized by sequence-diverse C-terminal regions (Rhs-CT and WapA-CT) that vary considerably between different strains of the same species. Analysis of several Rhs-CTs and WapA-CTs from Dickeya dadantii 3937 and Bacillus subtilis subspecies revealed that these domains contain the toxin activities responsible for intercellular growth inhibition. All rhs and wapA genes are closely linked to small downstream open reading frames that encode RhsI and WapI immunity proteins, respectively. These immunity proteins are also sequence-diverse and only protect against their cognate Rhs-CT (or WapA-CT) toxins. Thus, Rhs and WapA represent related, yet distinct, delivery platforms for polymorphic toxin domains [2]. Because different strains typically express unique rhs-CT/rhsI (wapA-CT/wapI) alleles, these systems collectively form a complex network of toxin/immunity pairs that are thought to mediate inter-strain competition for environmental resources [2]. The rhs loci of Enterobacteriacae often contain one or more additional rhs-CT/rhsI gene pairs located downstream of the main rhs/rhsI pair. These modules have been termed “orphan” toxin/immunity pairs, because the rhs-CT coding sequences resemble displaced fragments from full-length rhs genes [6]. Orphan rhs-CT genes often contain some coding sequence for portions of the conserved N-terminal regions, but orphan fragments are much smaller than full rhs genes and usually lack translation initiation signals. Therefore, it is unclear whether orphan rhs-CT genes are expressed, raising the question of whether these auxiliary elements are functional. Here, we show that repeated passage of Salmonella enterica serovar Typhimurium LT2 (StLT2) produces “evolved” lineages that deploy the orphan Rhs-CT toxin to inhibit the growth of ancestral cells. We provide evidence that the rhs locus undergoes rearrangement to fuse the rhsmain and rhs-CTorphan genes, thereby providing a mechanism to express and export the Rhs-CTorphan toxin domain. These results indicate that rhs rearrangement provides a selective advantage to a subpopulation of cells, suggesting that rhs plays an important role in clonal selection and bacterial evolution. In an effort to isolate StLT2 strains with increased fitness, we serially passaged cells for ∼1,000 generations in LB medium [7]. Analysis of six independently evolved cultures revealed that each lineage outcompeted ancestral StLT2 cells in co-culture experiments (Figures 1A & S1A). Remarkably, we observed the same competitive advantage in four of eight StLT2 lineages that were obtained by passage through multiple mouse hosts [8] (Figures 1A & S1B). This competitive advantage was not due to faster growth rate, because four of the evolved lineages grew more slowly than the ancestral strain (Figure S2). To further explore this phenotype, we tested whether evolved lineages inhibit ancestral cells in a contact-dependent manner. We co-cultured evolved and ancestral cells using trans-well culture dishes, in which the two populations are separated by membranes of different porosities [9]. The growth of ancestral cells was inhibited when the populations were separated by a cell-permeable 8. 0 µm filter, but not when cell contact was prevented with a 0. 4 µm filter (Figure 1B). These results indicate that evolved cells must be in close proximity to target cells in order to inhibit growth. This phenomenon is reminiscent of Rhs-mediated growth inhibition, which we recently characterized for D. dadantii 3937 [2]. StLT2 contains a single rhs locus, which contains a full-length “main” rhs gene (STM0291) and an “orphan” rhs gene fragment (STM0292) (Figure 2). Both rhs genes are closely linked to small open reading frames representing potential rhsI immunity genes (Figure 2), although the predicted rhsImain immunity gene found downstream of rhsmain is not annotated in the genome sequence NC_003197. To determine if the rhs region is responsible for the observed growth inhibition, we tested whether over-expression of either rhsImain or rhsIorphan immunity genes provided protection against evolved StLT2 lineages. Parental StLT2 cells overexpressing rhsImain were still inhibited by the evolved lineages, but overexpression of the rhsIorphan gene fully protected targets from growth inhibition (Figure 1A). These data strongly suggest that evolved StLT2 cells gained the ability to deliver Rhs-CTorphan toxin into neighboring cells. We next tested each rhs/rhsI gene pair to confirm that they encode functional toxin and immunity proteins. Nucleotides 3608 to 4095 of rhsmain and nucleotides 269 to 741 of rhs-CTorphan were cloned under the control of the arabinose-inducible PBAD promoter. The predicted rhsI immunity genes were cloned using a compatible plasmid under control of the IPTG-inducible Ptrc promoter. These plasmids were then introduced into StLT2 cells to evaluate toxin and immunity functions. Induction of either rhs-CTmain or rhs-CTorphan in StLT2 resulted in rapid growth arrest (Figure 3A). In each instance, growth inhibition was neutralized by expression of the cognate rhsI immunity gene. However, co-expression of non-cognate immunity genes did not alleviate growth arrest (Figure 3A), demonstrating that RhsImain and RhsIorphan immunity proteins are specific for their cognate toxins. We obtained essentially identical results upon expressing the rhs main toxin and immunity genes in E. coli cells (Figure 3B). These results indicate that Rhs-CTorphan is capable of inhibiting bacterial growth and support a model in which evolved StLT2 lineages deploy the orphan toxin to inhibit the ancestral strain. The rhs-CTorphan sequence does not encode a full-length Rhs protein, raising the question of how this toxin is synthesized and exported from evolved cells. The rhsmain and rhs-CTorphan coding regions share 95% sequence identity over 522 base-pairs (Figure 2), raising the possibility that homologous recombination in the evolved lines generates a new full-length rhs gene that encodes the Rhs-CTorphan toxin domain [10]. Bacteria expressing this Rhs chimera would have a growth advantage if rhsIorphan expression is low in ancestral cells. However, the proposed recombination event would also delete the rhsImain gene, rendering the evolved cells sensitive to inhibition by siblings expressing the main Rhs-CT toxin. Therefore, we hypothesized that rhs recombination occurs subsequent to duplication of the locus such that evolved cells retain the rhsImain immunity gene (Figure 2). To test this hypothesis, we analyzed chromosomal DNA from evolved and ancestral lineages by Southern blot. DNA was digested with HincII, which cleaves between the rhsImain and rhs-CTorphan coding sequences, and probed with a labeled DNA fragment that specifically hybridizes to rhsmain (Figure 2). We detected a unique junction fragment representing fusion of rhs-CTorphan to the upstream rhsmain gene in StLT2 lineage 2, which displayed the highest level of growth inhibition of all lineages (Figures 1 & 4A). The wild-type rhs locus was also detected in lineage 2 (Figure 4A), which is consistent with rhs region amplification, but may also indicate distinct populations of recombinant and non-recombinant cells. Orphan rhs recombinants were not detected in the other evolved lineages by Southern blot analysis (Figure 4A). Because the growth inhibition phenotype varied in magnitude between the different evolved strains, it is possible that only a fraction of the evolved StLT2 cells are rhs recombinants. If so, then the proportion of recombined rhs loci in the DNA sample may be below the detection limit of Southern analysis. Therefore we analyzed each evolved lineage with quantitative real-time PCR (qPCR) to measure the relative levels of rhsmain-rhsorphan junction sequences. All five of the evolved lineages contained 10- to 1,000-fold more rhsmain-rhsorphan junction than ancestral StLT2 (Figure 4B), consistent with the ability of these strains to deploy Rhs-CTorphan toxin. Because only a fraction of the passaged cells appeared to display growth inhibitory activity, we asked whether inhibitor-cell clones could be isolated from each population. As a control, we first isolated colonies from an overnight culture of the ancestral strain and tested these clones for growth inhibition activity. None of the ten ancestral clones tested were inhibitory, suggesting that the proposed rhs rearrangements occur at low frequency. By contrast, approximately 30–90% of the clones isolated from the culture-evolved lineages and ∼20% of the clones from mouse-evolved lineage 1 showed inhibition activity against ancestral cells (Figure 5A). However, no inhibitor clones were isolated from mouse-evolved lineage 2 (Figure 5A). Strikingly, the inhibition activity of these clones varied considerably. For example, competitive index values ranged from 10−1 to 10−5 for competitions between ancestral cells and inhibitory clones isolated from evolved lineage 2 (Figure S3). Although their potencies varied, it appears that each inhibitor-cell clone deployed the Rhs-CTorphan toxin because ancestral cells could be protected through over-expression of rhsIorphan, but not rhsImain (Figure 5B). The presence of DNA fragments corresponding to both ancestral and recombinant rhs loci in lineage 2 (Figure 4A) suggests that either the rhs region was duplicated or there are distinct populations of recombinant and non-recombinant cells. In the latter case, single colonies isolated from the inhibitory lineages would contain only the rhs- rhs-CTorphan junction and not the rhs-CTmain sequence. However, PCR analysis of the single colonies with inhibitory activity in Figure 5B showed that each contained both ancestral and recombinant rhs loci. In addition, sequence analysis of the recombinant PCR product verified that recombination occurred between the regions of homology shared by rhsmain and rhs-CTorphan. Together, these data demonstrate that the evolved populations are heterogeneous with respect to Rhs-CTorphan mediated inhibition activity. Furthermore, these results suggest that the inhibition phenotype of a given culture may be due entirely to a minor subpopulation of potent inhibitor cells. Because inhibitor cells represent a subpopulation in the evolved cultures, the other non-recombinant cells in the cohort are presumably resistant to the Rhs-CTorphan toxin. To test this hypothesis, we isolated non-inhibitory clones from each of the evolved cultures and tested them in competition co-cultures against their respective evolved lineages. As predicted, each of the non-inhibitory clones was either fully- or partially-resistant to its cohort lineage (Figure 5C). These cells likely carry uncharacterized resistance mutations that may prevent cell-cell contact, block the delivery of Rhs-CTorphan toxin,, or increase the immunity of these cells to Rhs-CT toxin. To directly detect Rhs-CTorphan expression in the evolved lineages, we examined cells by immunofluorescence microscopy using polyclonal antibodies against the Rhs-CTorphan toxin. Rhs-CTorphan antigen was detected on the surface of some cells within evolved lineages 1,2 and 3 as well as mouse-evolved lineages 1 and 2 (Figures 6A & S4). In contrast, the Rhs-CTorphan signal was undetectable on the surface of both ancestral StLT2 cells and cells carrying a deletion of the rhs-CTorphan (Figures 6A & S4). We then quantified the fraction of cells with Rhs-CTorphan antigen on the cell-surface using flow-cytometry. Evolved lineages showed a 2- to 20-fold increase in the fraction of Rhs-CTorphan-positive cells compared to ancestral StLT2 cells (Figures 6B & S5). Mouse-evolved StLT2 showed very low expression of Rhs-CTorphan antigen on cell surfaces (Figure 6B), consistent with the modest growth inhibition observed for these lineages (Figure 1). Based on Southern blot and RT-qPCR analyses, it seems likely that surface expression of Rhs-CTorphan requires rhs locus rearrangement to generate a chimeric rhs gene. In accord with this conclusion, we also found that over-expression of rhs-CTorphan from a multicopy plasmid does not increase Rhs-CTorphan antigen levels on the cell surface (Figures 6B & S5). Therefore, we sought to detect the predicted Rhs fusion protein using antisera to the Rhs-CTorphan toxin. Western blot analysis revealed an immuno-reactive protein at ∼150 kDa in culture evolved lineages 2 and 3 (Figure 6C). This product corresponds to the expected size of the Rhs fusion protein. Moreover, we were unable to detect the 29 kDa product encoded by rhs-CTorphan in the ancestral and evolved lineages (Figure 6C). Together, these data strongly suggest that the rhs-CTorphan reading frame must recombine with rhsmain to be expressed. The results presented here show that serial passage of StLT2, in either laboratory media or within a natural host, leads to enrichment of cells that express Rhs-CTorphan toxin. Analysis of the rhs locus indicates that evolved cells undergo recombination between rhsmain and rhs-CTorphan, forming a gene fusion that allows the Rhs-CTorphan toxin domain to be deployed. Rearranged rhs genes are detected at low levels within the evolved populations, indicating that only a fraction of cells are recombinant inhibitors. A number of observations argue that this subpopulation of cells is responsible for growth inhibition activity. First, the relative competitive advantage of each evolved lineage is correlated with its level of recombinant rhs junctions and surface expression of Rhs-CTorphan antigen. More importantly, ancestral cells are fully protected when they over-express the rhsIorphan immunity gene. Because Rhs immunity proteins are highly specific for their cognate toxins, this latter result demonstrates that Rhs-CTorphan toxin is indeed deployed by the evolved lineages. This result also indicates that ancestral StLT2 cells do not normally express rhsIorphan immunity genes under laboratory conditions. The number of inhibitor cells within each lineage is not known, but can be estimated to be <2% of the population based on flow cytometry measurements of Rhs-CTorphan antigen on cell surfaces. However, we note that this assay may underestimate the actual number of recombinant inhibitor cells because Rhs effectors are likely exported through type VI secretion systems [2], [11]. Although recent studies indicate that the N-terminal PAAR domain found within many Rhs proteins forms the tip of the type VI injection structure [12], other structural studies show that Rhs-peptide repeats form a chamber capable of encapsulating toxin domains [13]. Therefore, much of the Rhs-CTorphan antigen may be inaccessible to antibody until it is delivered to target cells. In accord with this model, we only detect Rhs-CTorphan where two bacteria make contact with one another and never on the surface of individual cells. Regardless of the absolute number of recombinants or rhs expression levels, our results suggest that a small number of inhibitor cells are capable of inhibiting a large excess of ancestral cells. The same phenomenon has been observed during bacterial contact-dependent growth inhibition (CDI), in which each CDI+ cell is able to inhibit 100–1,000 target cells over a few hours [9]. Presumably, the unstructured environment in shaking broth culture promotes a series of transient cell-cell interactions, thereby enabling toxin delivery to multiple ancestral cells. Chromosomal duplications and amplifications occur frequently in bacteria, typically at rates of about 0. 1% per generation for any given locus [14], [15]. However, there is a cost to maintaining amplified regions, and gene duplications are lost during segregation at frequencies up to 10% per generation [16], [17]. Therefore, positive selection is required to retain multiple gene copies. If the amplified region can be stabilized, then the additional gene copy can diverge towards a new function, thus providing a mechanism for evolution [16], [18]. Rearrangement of rhs loci represents a previously unrecognized mechanism for bacteria to exploit chromosomal amplifications for adaptation. We propose that, subsequent to duplication, homologous recombination occurs between rhsmain and rhs-CTorphan to generate a novel chimeric rhs element. This recombination would necessarily delete one copy of the rhsImain, but the other copy would remain and ensure that recombinant cells retain immunity to the Rhsmain toxin should it be deployed by neighboring non-recombinant siblings. This model also predicts that evolved recombinant cells could undergo homologous recombination to restore the original rhs locus (see Figure 2, reverse of the duplication step). Thus, rhs rearrangement could be exploited transiently under conditions where it confers a selective advantage, but rapidly revert back to the ancestral genotype as environmental circumstances dictate. Analysis of over 150 Salmonella genomes shows that rhs-CT toxin sequences are diverse with at least 57 distinct sequence types (Figure S6A & Table S1). This is a common feature of rhs genes in other bacteria as well and suggests that Rhs mediates inter-strain competition. All Salmonella serovars contain at least one rhs gene, located on pathogenicity islands SPI-6 or SPI-19 [19], [20]. Approximately 50% of these serovars contain at least one predicted rhs orphan sequence, with some strains containing as many as eleven modules. There is generally high conservation of Rhs-CTmain and Rhs-CTorphan. sequences within a given serotype. For example, all sequenced Typhi isolates contain the same Rhs-CTmain and Rhs-CTorphan. sequences, whereas these CT sequence types are only found in one other serotype, thus suggesting that different toxins are linked to serotype and/or the type of infection. However, orphan rhs-CT sequences in one serotype can be present within the main rhs gene of another serotype. For example, the StLT2 Rhs-CTorphan toxin studied in this work is part of the full-length main Rhs in Salmonella enterica serovar Saintpaul SARA23 and some Newport isolates (Figures S6A & S6B). These observations and the association with horizontally transferred elements suggest that rhs genes are exchanged between different serovars and contribute to the evolution of toxin diversity. Given that Rhs toxins are encoded on pathogenicity islands, it seems likely that these systems also play important roles in Salmonella growth and fitness during pathogenesis. Indeed, StLT2 mutants lacking a chromosomal region containing rhs-CTorphan are outcompeted by wild-type cells in mice [21], and StSL1344 mutants lacking rhs are completely attenuated in pig and cattle models of infection [22]. These observations raise the possibility that rhs locus rearrangement occurs commonly during infections. Intriguingly, StLT2 produces distinct intracellular infection foci, each originating from one or only a few clones [23]. Similarly, analysis of mice orally infected with Yersinia pseudotuberculosis indicates that only a few bacterial clones are able to disseminate from the intestines to the spleen and liver [24]. Clonal invasion has also been reported for Yersinia enterocolitica infections [25], but the mechanisms underlying these apparent dissemination bottlenecks are unknown. Most Yersinia species contain rhs loci with associated orphan gene pairs, raising the possibility that clonal expansion through rhs recombination and growth selection may be a general feature of many enterobacterial infections. Rearrangement could function as a stochastic switch that enables some cells to deploy Rhs-CTorphan and thereby “differentiate” into cells that are specialized for tissue invasion or immune modulation. Although Rhs-mediated inhibition clearly occurs between bacteria, it is also possible that Rhs toxins act directly as virulence factors. The C-terminal region of RhsT from Pseudomonas aeruginosa was recently shown to be delivered into mouse host cells [26]. In the process, the Rhs fragment activates the inflammasome and contributes to pathogenicity. Bacterial strains were derived from Salmonella enterica serovar Typhimurium LT2 (StLT2) and are listed in Table S2. Bacteria were grown in LB medium [27] supplemented with 50 mM potassium phosphate (pH 7. 3). Bacteria were incubated at 37°C with shaking at 200 rpm. Where appropriate, media were supplemented with antibiotics at the following concentrations: ampicillin (Amp), 200 mg/L; chloramphenicol (Cam), 17 mg/L; kanamycin (Kan), 80 mg/L; and tetracycline (Tet), 5 mg/L. Six independent lineages of StLT2 were obtained by serial passage for ∼1,000 generations [7]. Each lineage was passaged daily by dilution of 1. 5 µL of overnight culture into 1. 5 mL of fresh LB medium. Each evolved lineage was sampled periodically (100–150 generations) and stored at −80°C. All growth competitions were conducted using ancestral StLT2 marked with the flhC: : cat allele, which confers Cam resistance. Non-inhibitory clones isolated from the evolved cultures were transduced with the flhC: : cat allele prior to testing for resistance. Ancestral and evolved cells were co-cultured in LB medium supplemented with 50 mM potassium phosphate (pH 7. 3) at 37°C with shaking. At time 0 h, ∼106 cfu (1 µL of overnight culture) from evolved and ancestral cultures were suspended in 2 mL of fresh LB (pH ∼7. 3) and plated for viable cell counts before shaking incubation for 24 h at 37°C. After 24 h of co-culture, viable cell counts were determined by plating onto LB agar (to enumerate evolved and ancestral cells) and LB agar supplemented with Cam (to enumerate ancestral cells). The competitive index was calculated as the ratio of ancestral∶evolved cells at time 24 h divided by the cell ratio at 0 h. Ancestral StLT2 flhC: : cat cells were also supplemented with either rhsImain or rhsIorphan on the chromosome under control of the lac promoter and on plasmid pBR322 under the tet promoter. Chromosomal rhsIorphan and plasmid-borne rhsIorphan individually provided partial protection against the evolved lineages (data not shown), but both copies were required for full immunity. Proximity-dependence of growth inhibition was determined as described previously [9]. Cells were grown to OD600 ∼0. 3, then transferred to a trans-well culture plate (BD diagnostics) that separates the two populations with filter containing 0. 4 µm (no-contact) or 8. 0 µm (contact) pores. Trans-well culture plates were seeded at an evolved∶ancestral cell ratio of 1∶1 and incubated at 37°C with shaking for 24 h. Cultures were then plated onto selective media to determine viable cell counts and to calculate competitive indices. All oligonucleotides used in this study are presented in Table S3. The rhsImain and rhsIorphan genes were amplified from ancestral StLT2 chromosomal DNA using oligonucleotides 2337/2338 and 2340/2544 (respectively) and ligated to plasmid pBR322 using EcoRV and SalI restriction sites. The immunity genes were also placed under the lac promoter at the glmS locus using bacteriophage λ Red-mediated recombination [28]. Integration constructs containing rhsI genes flanked by a Kan-resistance cassette and glmS-derived homology regions were constructed by overlapping end-PCR as described previously [29]. The following primer pairs were used to amplify: upstream glmS homology (2666/2676), lac promoter (2677/2678 for rhsImain and 2677/2682 for rhsIorphan), rhsImain (2679/2680) or rhsIorphan (2683/2684), Kan-resistance cassette (2618/2619) and downstream glmS homology (2681/2667). The final PCR product was electroporated into StLT2 cells that express Red recombinase proteins, and transformants were selected on LB supplemented with Kan. Integrated immunity genes were verified by PCR analysis using primers 2666/2667 and subsequent DNA sequencing. The flhC: : cat and STM0292: : kan alleles were generated by PCR using primers 2436/2437 and 2410/2490 to amplify the cat/kan cassettes of plasmids pKD3 and pKD4, respectively. Each PCR product was integrated into the StLT2 chromosome by Red-mediated recombination. To evaluate toxin activity and the specificity of immunity, individual rhs-CT and rhsI sequences were cloned under the control of inducible promoters on compatible plasmids. The rhs-CTmain and rhs-CTorphan coding sequences were amplified with primers Sty-rhs (E1203) -Nco/Sty-rhs-Xho and Sty-rhs (E1203) -Nco/Sty-orph-rhs-Xho (respectively) and ligated to plasmid pCH450 [30] using NcoI and XhoI restriction sites. The rhsImain and rhsIorphan genes were amplified and ligated to a derivative of plasmid pTrc99A using KpnI and XhoI restriction sites. Rhs-CTorphan was expressed and purified as a non-toxic variant fused to His6-tagged thioredoxin. The his6-trxA sequence was amplified from plasmid pSH21P: : trxA [31] using primers pET-Sph and trxA-Bam-TEV-Kpn. The product was digested with SphI/BamHI and ligated to plasmid pET21b to generate plasmid pSH21P: : trxA-TEV. The coding sequences for Rhs-CTorphan residues 112–247 and RhsIorphan were amplified using primers Sty-rhs (D1225) -Kpn/Sty-orph-rhsI-Xho) and the His208Ala mutation made by mega-primer PCR using oligonucleotide Sty-CTo1-H208A. The final product was digested with KpnI/XhoI and ligated to plasmid pSH21P: : trxA-TEV to generate plasmid pCH10068. The resulting construct was used to overproduce His6-TrxA-Rhs-CT (H208A) orphan fusion protein. Chromosomal DNAs were isolated using the Sigma genomic DNA kit and digested with HincII restriction endonuclease. Digested DNAs were resolved by electrophoresis on 0. 7% agarose gels at 34V for 15 h and blotted onto nylon membranes by capillary transfer. A probe to nucleotides 2969–3128 of rhsmain was generated by PCR using oligos 2226/2227 and labeled with [32P]-labeled using the Prime-It Random Primer Labeling Kit (Agilent Technologies). Southern blots were visualized by phosphor imaging. Fragment sizes were calculated using a standard curve based on HindIII digested λ ladder (New England Biolabs, USA) run on the same gel. The proportion of rhs recombination junctions was determined by quantitative real-time PCR (qPCR) using oligonucleotides 2226/2231 using the cycle threshold Ct-value method according to the manufacturer (Bio-Rad). Fluorescence was monitored on-line using the MyIQ iCycler real-time PCR system (Bio-Rad). The rhs-rhsorphan junction DNA levels were calculated relative to bamA DNA (oligos 1981/1990) in each sample and normalized to the level of junction DNA in ancestral cells. His6-TrxA-Rhs-CT (H208A) orphan fusion protein was overproduced in E. coli CH2016 and purified by Ni2+-affinity chromatography as described [32]. The Rhs-CT (H208A) orphan domain was released by TEV protease digestion and used for antiserum production in rabbits (CoCalico Biologicals). Non-specific antibodies were removed by incubation with carbonyl-diimidazole-activated agarose beads linked to soluble protein from E. coli strain CH2016 [33]. Briefly, protein-linked beads were resuspended in 0. 5 mL of antiserum (1∶5 dilution) and mixed by rotation for 1 h at room temperature followed by additional incubation for 3 h at 4°C. This process was repeated at least four times with fresh beads. Evolved lineages were grown to mid-log phase in LB medium supplemented with 50 mM potassium phosphate (pH 7. 3) and cells were collected by centrifugation and frozen at −80°C. Cell pellets were resuspended in NuPage LDS-sample buffer (Invitrogen) at 70°C and treated with benzonase to degrade nucleic acids. Cell lysates were run on 3–7% NuPage Tris-acetate gradient gels (Novex) for the detection of the Rhsmain-Rhs-CTorphan chimera, or on 4–10% Precise Tris-glycine gradient gels (Thermo Scientific) to detect Rhs-CTorphan. Gels were electrotransferred to nitrocellulose membranes and the blots incubated with polyclonal antisera against Rhs-CTorphan (1∶1,000 dilution) and secondary anti-rabbit 800CW antiserum (1∶10,000 dilution). Immunoblots were visualized using an Odyssey CLx Infrared Imaging System (LI-COR). Cells were incubated overnight with 4% formaldehyde in 0. 15 M phosphate buffered saline (PBS, pH = 7. 2). Cells were washed three times with PBS and incubated with polyclonal antibodies to Rhs-CTorphan (1∶50 dilution) for 30 min. Cells were washed with PBS before incubation with secondary anti-rabbit Alexa-Fluor480 antibodies (1∶500 dilution) (Invitrogen) for 30 min on ice. After washing with PBS, cells were applied to poly-D-lysine coated slides, treated with Fluoro-gel II/DAPI (Electron Microscopy Sciences) and visualized by fluorescence microscopy. The fraction of evolved cells expressing Rhs-CTorphan on the surface was determined by flow cytometry. Antibody-labeled cells were analyzed (50,000 events for each sample) with an Accuri C6 flow cytometer with gates set to include bacteria-sized particles. StLT2 Δrhs-CTorphan cells were used to assess non-specific binding of the Rhs-CTorphan antisera. The fraction of cells with surface Rhs-CTorphan antigen was calculated as the ratio of green fluorescent particles in the population after subtracting background fluorescence observed with StLT2 Δrhs-CTorphan cells.

Title: Selection of Orphan Rhs Toxin Expression in Evolved Salmonella enterica Serovar Typhimurium Summary: Salmonella Typhimurium is a bacterium that causes intestinal diseases in a number of animals including humans. In mice, this pathogen invades tissues, causing symptoms similar to typhoid fever. In an effort to understand the evolution of this pathogen, we grew S. Typhimurium in either liquid broth or in mice for many generations and examined the resulting "evolved" strains to determine if they were different from the original "parent" culture. We found that many of these evolved strains inhibited the growth of the parent after they were mixed together, and that this growth inhibition requires that the evolved and parental cells are in close contact. Genetic analysis showed that this contact-dependent growth inhibition requires Rhs protein, which has a toxic tip. Salmonella is normally resistant to its Rhs toxin because it also produces an immunity protein that blocks toxin activity. However, evolved cells have undergone a DNA rearrangement that allows them to express a different Rhs toxic tip that inhibits growth of the parental cells, which lack immunity to it. This allows the evolved cells to outgrow the original parental cells. Our work indicates that populations of Salmonella are dynamic, with individuals battling with each other for dominance.

lay_plos

en

Summarize: By adding a special protein, researchers say, they can make your ice cream cone drip-free. (Yue Wu/The Washington Post) Scientists claim to have found a solution for dripping ice cream cones: a protein that keeps the treat from melting. The protein, called BslA (Bacterial Surface Layer A), is already found in other foods, like the Japanese breakfast food of fermented soybeans called natto. When the bacteria Bacillus subtilis grows into colonies called biofilms, the colony protects itself by forming a coating of BslA, which forms a kind of "bacterial raincoat." Cait MacPhee of the University of Edinburgh, who's leading the research on the protein's potential uses, realized that the physical properties of this bacterial raincoat might make it perfect for ice cream. The protein, she observed, could coat air bubbles and oil droplets in the same way it coated the bacterial colonies, as well as coating solid surfaces. [The science behind perfect grilling] "Bring those three together and you’ve got ice-cream," MacPhee told The Post in an e-mail, "an oil + water (or sugar syrup) mixture, plus air bubbles, plus ice crystals (the solid). So if we add the protein it can protect all three, and keep the mixture stable." In addition to keeping the ice cream more stable -- which means it doesn't easily melt in the traditional sense, though it does get warm (and therefore less delicious) over time -- the protein also keeps ice crystals from forming on the dessert. That keeps the texture nice and smooth, unlike traditional ice cream that gets hard and frosty in the freezer. [Why nuking frozen veggies might be healthier than poaching fresh greens] MacPhee and her colleagues have already published two papers on the way the protein works -- one focusing on its biology and the other on its physics -- and she says they've already tested the ice cream application in the lab. With a patent pending, however, her team is keeping a tight lid on the data they collected. So we'll have to wait and see just how effective this anti-melting agent really is. If the protein is successful, MacPhee and her colleagues believe it could improve ice cream all up and down the supply chain. It would take less energy to make the product, since it didn't have to be as cold, and less energy to transport it for the same reason. Once in the hands of a consumer, the ice cream would theoretically stay nice and creamy long past when other products would have freezer burn, and of course the experience of eating it on a hot day would be greatly improved -- and much less sticky. Read More: Here’s what it takes to be the perfect poop donor These caffeine-addicted beetles are using bacteria to ruin all of our coffee Scientists have created the tear-less onion of your dreams Twitter can tell which states love jogging and which are eating hot dogs New research might teach us how to make a scarier scream Reduced-fat ice cream that doesn't melt? That's what U.K. scientists are said to be working on. A team of scientists from the University of Edinburgh and the University of Dundee said in a written statement that they have discovered a type of protein which could be used to create ice cream that is more resistant to melting. "The protein binds together the air, fat and water in ice cream, creating a super-smooth consistency," the scientists said in the statement, enabling summer treats to keep frozen for longer in hot weather. “We’re excited by the potential this new ingredient has for improving ice cream, both for consumers and for manufacturers," Professor Cait MacPhee, of the University of Edinburgh’s School of Physics and Astronomy, who led the project, said. Researchers estimate that ice cream made with the naturally occurring protein, known as BslA, could be available within three to five years. In addition, products manufactured with that protein would contain lower levels of saturated fat and fewer calories than those currently on sale. EDINBURGH, Scotland, Aug. 31 (UPI) -- Eating an ice cream cone on a hot afternoon is a race against time. But soon, scoop lovers may be able to savor their favorite treat, despite the heat. Researchers have discovered a protein that makes ice cream more resistant to heat, slowing the melting process. The protein, called BslA (Bacterial Surface Layer A), could also enable food scientists to create products with few saturated fats (and fewer calories). The protein is found naturally in and on colonies of the friendly bacteria Bacillus subtilis. It serves as a raincoat-like film, protecting the bacteria from the elements. BslA is present in foods that naturally contain the bacteria, including a fermented soybean product called natto, which is eaten for breakfast in Japan. Researchers in Scotland, at the University of Edinburgh and Dundee University, have shown that the bacterial biofilm could also be used to protect and bind the three elements that form ice cream -- air, fat and ice. "We're excited by the potential this new ingredient has for improving ice cream, both for consumers and for manufacturers," Cait MacPhee, a material scientist at Edinburgh, said in a press release. Just as the protein protects the bacterial colonies, it binds to the fat droplets and ice bubbles in ice cream, keeping the mixture more stable and less vulnerable to heat. "It has been fun working on the applied use of a protein that was initially identified due to its practical purpose in bacteria," said Nicola Stanley-Wall, a researcher at the University of Dundee. Scientists say the technology could be in the freezer aisles of local grocers in three to five years. Until then, grab extra napkins. One of the hazards of eating ice cream is having it melt all over the place, coating your fingers with sticky goo and forcing you to scramble for a pile of napkins. But scientists in Scotland may have come up with a way to keep that from happening. They have identified a naturally occurring protein that allows ice cream to last longer in the heat. The protein also has been shown to reduce those annoying ice crystals that form when ice cream sits in the freezer a little too long - ensuring a fine, smooth texture like those of luxury ice creams. "We are predicting that you should be able to eat an ice cream cone without the ice cream dribbling down the side," University of Edinburgh's Cait MacPhee, who led the team that worked out how the protein function, told CBS News. The protein, known as BslA, is already used in some Southeast Asian foods including a traditional soybean breakfast dish called Natto in Japan. It works by adhering to fat droplets and air bubbles, making them more stable in a mixture and preventing the ice cream from melting. MacPhee said the protein is formed as part of an effort by microbial communities to protect themselves. "They protect themselves by producing this protein and that protein goes to the outer surface of this community and makes a film that we dubbed a bacterial raincoat - it becomes basically water repellent," she said. "That means if there are any other bugs in the environment that want to attack our friendly bacteria, they can't get through because they bounce off. It's a pretty clever strategy." When she and Nicola Stanley-Wall of the University of Dundee first began studying the microbial communities that produce this protein, MacPhee admitted that making a better ice cream wasn't on their agenda. She was "mainly trying to characterize how well the protein behaved." But they soon realized much of what the protein does in terms of forming this bacterial raincoat or film would lend itself to prolonging the life of ice cream. "This film forms because there is an interface between the colony, which is wet, and the outside environment, which is air, essentially," she said. "So it can form this film between the interface of air and water. That instantly suggests it can stabilize air bubbles, which it does do. It also can stabilize a mixture of oil and water in exactly the same way. It also can coat solid surfaces," she said. "That combination of the three - having solid surfaces, having air bubbles and having oil and water mixtures - is the definition of ice cream." Guided by that concept, the researchers turned their lab into a mini Baskin-Robbins to see if indeed the protein worked. They tried their hand at making vanilla ice cream, replacing the emulsifier with their protein. "It melts more slowly, which was the big finding, but the ice crystals - you know when you have ice cream in your freezer for any extended time and you can get a gritty sensation in your mouth - it slows that down as well because the ice crystals can't grow as quickly," MacPhee said. But what about the big question: How does it taste? MacPhee acknowledged they have not yet tried eating their laboratory's ice cream, though she thinks wouldn't taste all that different. "We haven't actually tasted it yet. But what we are replacing is a small molecule that is there in a small amount," she said. "So it shouldn't have an impact on the taste because there is very little of it in there at all. There won't be any impact on the way it feels in your mouth either because the structure is the same." MacPhee, who is expecting to publish her findings soon, already has filed an application for a patent on the protein and is talking to food companies about its use in ice cream as well as other products ranging from salad dressing to chocolate mousse. The researchers are expecting that ice cream with this protein could be on supermarket shelves in three to five years.

Summary: Soon, you may be able to enjoy an ice cream cone without having to worry about sticky fingers. Scientists from the University of Edinburgh and the University of Dundee have discovered that a naturally occurring protein, known as BslA, can be used to make melt-resistant ice cream, ABC News reports. BslA (or Bacterial Surface Layer A, as UPI notes), which is already used in some Asian dishes, is created by microbial communities as they seek to protect themselves; it "goes to the outer surface of this community and makes a film that we dubbed a bacterial raincoat-it becomes basically water repellent," research leader Cait MacPhee tells CBS News. The protein adheres to air bubbles and droplets of fat, stabilizing them, and in the process protecting the microbial community: "If there are any other bugs in the environment that want to attack our friendly bacteria, they can't get through because they bounce off," MacPhee says. And then there's the ice cream-related side effect. Since the protein can protect and bind ice cream's three main ingredients (air, fat, and ice), "you should be able to eat an ice cream cone without the ice cream dribbling down the side" if it's used to make the frosty treat, says MacPhee, though, as the Washington Post notes, the ice cream does still get warm eventually. The team tested out the theory, replacing the emulsifier in a vanilla ice cream recipe with the protein, and found the resulting dessert did in fact melt more slowly. And, as the scientists note in a press release, there's an added bonus: "The protein binds together the air, fat, and water in ice cream, creating a super-smooth consistency"-which also means there won't be as many ice crystals formed in your ice cream if you leave it in the freezer too long. As for whether it's any good, the team didn't actually eat the creation-but since such a small amount of the protein is used, they don't think there will be any impact on the taste. When can you buy it? Researchers estimate it could be on sale within three to five years.

multi_news

en

Summarize: This application is a divisional of application Ser. No. 07/555,989, filed July 20, 1990 now U.S. Pat. No. 5,056,779. FIELD OF THE INVENTION This invention relates to torso exercise machines, and more particularly to an exercise machine having an upper body engaging member moveable along an eccentric path of travel for receiving force from a body part of a user and a range limiter for varying the starting position of the upper body engaging member and thereby reducing the range of movement along the path of travel. BACKGROUND OF THE INVENTION On most prior art exercise machines, when exercising the torso muscles such as the abdominal and lower back muscles, the user engages an upper body engaging member of the machine and exerts back and forth force thereagainst so that the spine of the user partially rotates around several vertebrae. During these back and forth movements, the axis of rotation of the user moves vertically along the spine. Heretofore, most prior art exercise machines for exercising the abdominal and lower back muscles have been constructed to include a body engaging member moveable in a back and forth semi-circular path of travel which is always the same distance from the rotational axis. For example, in commonly assigned U.S. Pat. Nos. 4,500,089 and 4,387,893, the body engaging member is mounted on the outer end of a user actuated lever. The inner end of the user actuated lever is pivotally connected to the machine frame so that the body engaging member is moved back and forth along a semi-circular path of travel. During the back and forth exercise movements, the lever rotates about a fixed axis of rotation. The maximum exercise efficiency is not obtained because the actual axis of rotation of the body moves vertically along the spine during the back and forth movements while the semi-circular movement of the body engaging member does not compensate for the movement of the rotational axis along the vertebrae of the spine. This prior art type of exercise machine causes rotation around the hips instead of the desired rotational movement along the spine. Additionally, in most prior art exercise machines such as disclosed in the aforementioned prior art patents, no means is provided for limiting the range of the body engaging member during the exercise. At times, it is desirable to vary the starting position of the body engaging member and thereby reduce its range of movement. For example, a back injury may necessitate adjusting the exercise machine so that instead of exercising the back with the full range of movement from a forwardly bent position to a rearwardly bent position, only a limited range of back movement is provided to prevent further injury. SUMMARY OF THE INVENTION Therefore, it is an object of the present invention to provide a torso exercise machine which overcomes the deficiencies of the prior art. Another object of the present invention is to provide a torso exercise machine which includes an upper body engaging member adapted for engaging a portion of the upper body of a seated user and receiving force from the upper body part for exercising an isolated portion of the torso, and wherein the upper body engaging member is moveable along a back and forth path of travel so that an instantaneous (moving) axis of rotation is generated substantially along the spinal column, corresponding to the changing axis of rotation of the isolated and exercised torso portion as the user applies back and forth force against the upper body engaging member. It is still another object of the present invention to provide a range limiter for use in an exercise machine of the aforementioned type for varying the starting position of the body and thereby reducing the range of movement of the body engaging member along the back and forth path of travel. These and other objects and advantages of the present invention are accomplished by a torso exercise machine which includes an upper body engaging member pivotally mounted about a horizontal pivot axis for back and forth movement along an eccentric path of travel. The eccentric path of travel of the upper body engaging member is obtained by a special pivotal mounting, illustrated as a four-bar linkage mechanism. This four-bar pivotal mounting means includes first and second lever arms having the first ends of each lever arm pivotally mounted in spaced relation to each other on the upper body engaging member. The second ends of the lever arms are pivotally mounted in spaced relation to each other on the frame so that the axis of rotation of the upper body engaging member changes as the upper body engaging member is moved back and forth along the eccentric path of travel. Resistance means, in the form of a stack of weight, is supported for vertical movement on the frame to provide resistance and oppose back and forth movement of the upper body engaging member by the user while positioned on the seat. Linkage means operatively connects the upper body engaging member to the resistance means for transmitting back and forth movement of the body engaging member to the resistance means. A variable radius cam is rotatable with the body engaging member and is operatively connected to the resistance means for varying the amount of force required to be exerted by the user on the body engaging member in accordance with the position of the body engaging member along the path of travel. Range limiter means is provided for varying the starting position of the body engaging member and reducing the range of back and forth movement along the path of travel. The range limiter means includes first sprocket means operatively connected to the body engaging member and second sprocket means operatively connected to the resistance means. Pawl means is provided to releasably lock the first sprocket means to the second sprocket means so as to permit the first and second sprocket means to be interconnected in a selected relative rotational orientation to thereby vary the starting position of the body engaging member along the back and forth path of travel. BRIEF DESCRIPTION OF THE DRAWINGS Some of the objects and advantages of the present invention having been stated, others will be more fully understood from the detailed description which follows and by reference to the accompanying drawings in which: FIG. 1 is a perspective view of the torso exercise machine in accordance with a first embodiment of the invention wherein the exercise machine is adapted for exercising abdominal muscles; FIG. 2 is an isometric view of the machine of FIG. 1, having the covers and padded portions removed from the frame and showing component parts of the machine; FIG. 3 is a plan view of the machine of FIG. 2, and having the padded seat included therewith; FIG. 4 is a side elevation view of the machine of FIG. 2, having the padded portions included; FIG. 5 is a rear elevation view of the machine of FIG. 2; FIG. 6 is an enlarged fragmentary elevational view of the range limiter in accordance with the present invention looking in the direction of arrow 6 of FIG. 5; FIG. 7 is a vertical sectional view of the range limiter, taken along line 7--7 of FIG. 6; FIG. 8 is a vertical sectional view of the range limiter, taken along line 8--8 of FIG. 6; FIG. 9 is a vertical sectional view of the range limiter, taken along line 9--9 of FIG. 8; FIG. 10 is a schematic representation showing the back and forth exercise movement of a user with the exercise machine of the first embodiment of FIG. 1, and showing the change of center of rotation generated by the lever arms; FIG. 11 is an isometric view similar to FIG. 2 but showing a second embodiment of the present invention wherein the exercise machine is adapted for exercising the muscles of the lower back; FIG. 12 is an enlarged isometric view of the lever arms and back engaging member, looking in the direction of arrow 12 of FIG. 11; FIG. 13 is a perspective view of the second embodiment of the present invention, similar to the view shown in FIG. 1; FIG. 14 is a plan view of the machine in accordance with the second embodiment, showing the cover in position on the machine; and FIG. 15 is a schematic view showing the back and forth movement of a user during exercise of the lower back. DESCRIPTION OF THE PREFERRED EMBODIMENTS Referring now to the drawings, and more particularly to FIGS. 1 through 10, there is illustrated a first embodiment of the torso exercise machine in accordance with the present invention which is adapted for exercising abdominal muscles. As best shown in FIG. 2, the exercise machine includes an upright frame, broadly indicated at 10. The upright frame 10 includes a continuous outer frame member 12 having a vertical rear leg, a horizontal upper part, and an inclined front leg. A lower horizontal frame member 14 is connected at opposite ends to the lower ends of the front and rear legs. A Vertical frame member 16 is fixed at its lower end to the lower horizontal frame member 14 and at its upper end to a first cross frame member 17. A second cross frame member 18 extends between the Vertical frame member 16 and the rear leg of the frame 10 and includes outwardly extending and spaced apart lateral support arms 20. A horizontal support member 21 connects the ends of the lateral support arms 20 to form a rectangular support structure extending on one side of the frame 10. Two spaced lower horizontal support members 24 extend laterally from the frame member 14 of the frame 10 and are connected to a seat frame, broadly indicated at 25. A vertical support frame member 22 is fixed at its upper end at the juncture of the support arm 20 and the support member 21 and at its lower end to the rear support member 24. The seat frame 25 includes a main L-shaped support member 26 having vertical and horizontal brace members 27 fixed thereto. A diagonal brace member 28 connects the frame 10 with the horizontal seat frame brace member 27 to provide rigidity. The L-shaped support member 26 includes a vertically adjustable seat mount 30 on which a padded seat cushion 31 is attached (FIG. 1). A padded back cushion 32 is attached to the upper end portion of the vertical portion of the L-shaped support member 26. Restraining means in the form of a seat belt 34 is attached to the L-shaped support member 26 to provide restraint to a user seated on the machine. Resistance means is supported for vertical movement on the frame 10 and includes a plurality of weight plates 36 which are supported for sliding movement on spaced guide rods 37 (FIG. 4). The upper ends of the guide rods 37 are fixed on the first cross frame member 17 and their lower ends are fixed on the lower horizontal frame member 14. A vertical selector guide and weight lifting rod 38 extends through the central portion of the weight plates 36. The weight plates 36 are provided with horizontal openings 40 for reception of a selector pin 41 so that varying amounts of weight can be selected by the user to be lifted and lowered when exercising with the machine. A two-piece molded cover 45 is mounted on opposite sides of the frame 10 and includes a slot opening 46 for gaining access to the selector pin 41. Body engaging means, in the form of an upper torso engaging member indicated generally at 50, is adapted for engaging at least the hands and arms of a user and receiving force therefrom for isolating and exercising abdominal muscles. The upper torso engaging member 50 includes a main support member 51 having vertically extending and spaced hand grips 52 extending upwardly and forwardly therefrom. Inclined elbow engaging pads 54 are supported on the forward ends of spaced tubular support braces 55. The upper torso engaging member 50 is mounted on the frame 10 for back and forth pivotal movement about a horizontal pivot axis which is positioned to generally pass through the spine of the user. The upper torso engaging means moves in an eccentric path of travel so that an instantaneous axis of rotation is generated substantially along the spine of the user corresponding to the changing axis of rotation of the abdominal muscles and spine as the user flexes against the upper body engaging member 50. Means is provided for pivotally mounting the upper torso engaging member 50 for movement along the eccentric path of travel and includes first and second lever arms 61 and 62 having first upper ends mounted in spaced relation to each other on a forwardly extending portion of the main support member 57, which forms a third lever arm 65. The third lever arm 65 interconnects the upper ends of the first and second lever arms 61 and 62. A lever sprocket 67 and sprocket hub 68 are rotatably mounted on the frame 10 between the horizontal support brace member 21 and the second cross frame member 18. The lower end of the first lever arm 61 is fixed to the sprocket hub 68 and is pivotable about the central axis of the lever sprocket 67. The lower end of the second lever arm 62 is pivotally mounted on the horizontal support brace member 21 on an axis of rotation extending in rearward spaced, parallel relation with the central axis of the lever sprocket 67 so that the center lines of the first and second lever arms 61 and 62 cross each other (FIGS. 4 and 10). A counterweight 69 is fixed to the sprocket hub 68 and opposite the first lever arm 61. As the user moves the upper torso engaging member 50 back and forth along its path of travel, the lever arms 61 and 62 generate an instantaneous center to provide a different axis of rotation along the abdominal muscles of the user positioned in the seat 31. The changing axis of rotation corresponds to the changing axis of rotation of the abdominal muscles of the user as the user flexes from a somewhat rearwardly bent position (shown in dashed lines in FIG. 10) to a substantially forward, crouched position (shown in dotted lines in FIG. 10). This type of described structure for changing the axis of rotation commonly is referred to as a four-bar linkage. A wide variety of structures which change the axis of rotation can be used with the present invention. The four-bar linkage generates an instantaneous center which is determined by the point at which the center lines of the first and second lever arms 61, 62 cross each other. In FIG. 10, the point at which the center lines cross each other (instantaneous center) is indicated at 62a, when the user is in the rearmost position. The point at which the center lines cross each other (instantaneous center) is indicated at 62b, when the user is in the forwardly crouched position. Thus, the instantaneous center moves downwardly and forwardly as the user bends forwardly and moves upwardly and rearwardly as the user bends rearwardly. A variable radius cam 70 is provided and is operatively connected between the lever sprocket 67 and the weight plates 36 to provide proper variable resistance for varying the amount of force required to be exerted by the user on the upper torso engaging member 50 in accordance with the position of the upper torso engaging member 50 along the eccentric path of travel. A chain 71 interconnects the front edge of the lever sprocket 67 With the variable radius cam 70. As illustrated in greater detail in FIGS. 6 through 8, the variable radius cam 70 is mounted for rotational movement on a bolt forming a central shaft 73. Opposite ends of the bolt shaft 73 are supported in respective outer and inner cross frame members 73a, 73b (FIGS. 7 and 8). Range limiter means, broadly indicated at 75, is also supported on the shaft 73 and is provided for varying the starting position of the torso engaging member 50 and to thereby reduce its range of movement along the eccentric path of travel. The range limiter means 75 includes a first double tooth sprocket 76 mounted for rotation on the shaft 73 and fixed to the variable radius cam 70, as by a bolt 74 (FIG. 8). A second larger sprocket 77 is mounted for rotational movement on the shaft 73 and is operatively connected by a chain 78 to the weight lifting rod 38 extending through the weight plates 36. The chain 78 passes over an idler sprocket 79 (FIG. 11) supported above the cross frame member 17. A pawl so is pivotally mounted to the second larger sprocket 77 and includes a latching tooth 81 dimensioned for receipt into the double row of teeth of the first sprocket 76. A tension spring 82 interconnects the pawl so and second sprocket 77 for biasing the pawl 80 into engagement with the teeth of the first sprocket 76. A hand lever, indicated generally at 83, is pivotally mounted to the frame 10 and includes a bar member 84 moveable with the hand lever for engaging an extension 85 of the pawl 80 for moving the pawl 80 out of engagement with the first sprocket 76 (FIG. 6). A spiral torsion spring 90 is mounted at one end, as indicated at 91 in FIG. 9, on the second sprocket 77 and includes a second end fixed on the first sprocket 76,as indicated at 92, for biasing the second sprocket 77 in a counterclockwise direction and to an initial starting position when the pawl 80 and latching tooth 81 is disengaged from the first sprocket 76. The initial starting position of the torso engaging member 50 can be adjusted by the seated user moving the handle 83 in a clockwise direction to raise the pawl 80 and release the latching tooth 81 from engagement with the teeth of the first double teeth sprocket 76. The user then moves the torso engaging member 50 rearwardly to the desired starting exercise position. The handle 83 is then released to permit the latching tooth 81 to again engage the teeth of the first double teeth sprocket 76. This action provides a reorientation of the rotational position of the first double teeth sprocket 76 relative to the rotational position of the second sprocket 77 to adjust the starting position of the exercise and to accordingly adjust the range of operation of the torso engaging member 50. Referring now to FIGS. 11-15, a second embodiment of the torso exercise machine of the present invention is illustrated which is constructed for exercising muscles of the lower back. Throughout the description of this second embodiment, the same reference numerals, with the prime notation added, will be used for corresponding elements described in the first embodiment of FIGS. 1-10. As illustrated, the seat frame 25&#39; includes a diagonal support bar 101 extending upwardly along a forward portion of the frame 10&#39;. A foot rest support 102 is mounted for vertical adjustment on the support bar 101 and has a foot pad 100 fixed thereto. A padded seat 103 and a padded lower back rest 104 are supported on the L-shaped support member 26&#39; (FIGS. 13 and 14). As illustrated in FIG. 12, the chain 71&#39; interconnecting the lever sprocket 67&#39; and variable radius cam 70&#39; interconnects the rear edge of the lever sprocket 67&#39; so that back engaging means, illustrated generally at 110, operates to lift the weight plates 36&#39; when the back engaging means 110 is moved rearwardly by the user. The back engaging means 110 includes a horizontal rod 111 having a back engaging pad 112 pivotally supported on the outer end portion by a collet 113. The inner end portion of the rod 111 is fixed in the forward end of a third lever arm 114 extending transverse to the horizontal rod 111. The back engaging means 110 also is mounted to the frame 10&#39; for pivotal back and forth movement about a horizontal pivot axis and along an eccentric path of travel. As in the first embodiment, first and second lever arms 61&#39; and 62&#39; support the back engaging means 110 so that an instantaneous axis of rotation is generated substantially along the spine of the user. The upper first ends of the lever arms 61&#39;, 62&#39; are pivotally connected to the third lever arm 114. The lower end of the second lever arm 62&#39; is pivotally connected to the horizontal support brace member 21&#39; on an axis of rotation extending forwardly of and in spaced, parallel relation to the central rotational axis of the sprocket hub 68&#39;. In this lower back exercising machine, the center lines of the first and second lever arms 61&#39;, 62&#39; of the four-bar linkage do not cross within the length of the lever arms 61&#39;, 62&#39;, as they did in the abdominal exercise machine of the first embodiment. Instead, the center lines of the lever arms 61&#39;, 62&#39; cross at a first instantaneous center 62a&#39; (FIG. 15) positioned below the lower pivot points of these lever arms 61&#39;, 62&#39; when the user is in the forward dashed line position. When the user moves the upper portion of the body rearwardly to the dotted lines position, the instantaneous center moves to a second position 62b&#39;, moving along the curved line indicated in FIG. 15 and connecting the first and second positions 62a&#39;, 62b&#39;. Method of Operation In the abdominal exercising machine of the first embodiment illustrated in FIGS. 1-10, the user is positioned in the seat 31 and straps the seat belt 34 across his upper thighs (FIG. 10). The user places his elbows on the elbow engaging pads 54 and grips the vertically extending hand grips 52, as shown in dashed lines in FIG. 10. If a full range of movement is not desired, such as in the case of user having an injury and needing rehabilitation, the range limiter hand lever 83 can be moved to disengage the latching tooth 81 of the pawl 80 from the first double teeth sprocket 76. The user then presses forwardly on the elbow engaging pads $4 and the hand grips 52 to move the upper torso engaging member 50 a predetermined distance along the eccentric path of travel to a desired starting position. The hand lever 83 is then released to engage the latching tooth 81 of the pawl 80 with the double row of teeth on the first sprocket 76 to reorient the rotational position of the variable cam 70 relating to the second sprocket 77. The user then selects the desired resistance by inserting the selector pin 41 into the desired opening 40 to engage the weight plates 36 with the weight lifting rod 38. The user pulls the upper torso engaging member 50 forwardly and moves from a first somewhat rearwardly bent position (as shown in dashed lines in FIG. 10) to a second position where the user is in a somewhat upright forward, crouched position (as shown in dotted lines in FIG. 10). The user repeats the cycle for as many repetitions as necessary for the exercise program. During the exercise movement the axis of movement and the instantaneous center changes as indicated by the double headed arrow in FIG. 10. In the second embodiment illustrated in FIGS. 11-15, the user straps himself in the seat as before. The desired weight is selected and the desired starting position is selected by moving the handle 83&#39; to disengage the pawl 80&#39; from the teeth of the first sprocket 76&#39; to vary the starting position of the back engaging means 110. During the exercise movement, the user presses against the back engaging means 110 to move between the first position with the spine of the user in a forwardly bent position, shown in dashed lines in FIG. 15, and a second position with the spine of the user in a rearwardly bent position, shown in dotted lines in FIG. 15. The user repeats the cycle for as many repetitions as necessary during the exercise program. The present invention offers several benefits over other prior art exercise machines. The structure of the lever arms generates an instantaneous (changing or moving) axis of rotation substantially along the vertebrae of the spine. This instantaneous center corresponds to the changing axis of rotation for the isolated and exercised torso portion, such as the lower back and abdominal muscles, as the user flexes against the upper body engaging means. Thus, exercise efficiency is increased and the possibility of injury during the exercise movement is lessened because there is rotation about the spine, and not the hip joint. Additionally, the range limiter means varies the starting position of the upper torso engaging means and reduces the range of movement along the path of travel. This especially is beneficial for those users which are rehabilitating old injuries where the full range of exercise movement is not desirable. In the drawings and specification, there has been set forth the best modes presently contemplated for the practice of the present invention, and although specific terms are employed, they are used in a descriptive sense only and not for purposes of limitation, the scope of the invention being defined in the claims.

Summary: A torso exercise machine includes a seat connected to a frame for supporting a user thereon. An upper body engaging member receives force from an upper body part for exercising the torso. The upper body engaging member is pivotally mountd to the frame for pivotal movement along an eccentric path of travel while generating an instantaneous axis of rotation substantially along the torso and spine of the seated user. The eccentric path of travel of the upper body engaging member is illustrated as being obtained by pivotally mounting the upper body engaging member to the frame by a four-bar linkage. A resistance weight opposes movement of the user while the user exercises. A range limiter is provided to reduce the normal range of back and forth movement of the upper body engaging member.

big_patent

en

Summarize: BACKGROUND OF THE INVENTION The present invention relates to a self-service terminal (SST). In particular, the invention relates to a public access self-service terminal such as an automated teller machine (ATM) or a non-cash kiosk. It is well known that ATMs are commonly used as a convenient source of cash and other financial transactions. Some users of ATMs desire a quick cash dispense transaction (sometimes referred to as fast cash) without viewing any promotional material (such as advertisements or marketing information) or other services (such as other transactions). Other users, however, are willing to view promotional material and/or services on ATMs depending on certain factors, such as whether they are in a hurry, whether they are interested in the type of product or service that is being promoted, or such like. If promotional material or services are presented to users at inappropriate times, or if an ATM transaction is lengthened because of presenting other services or soliciting some input from the user, then the user may be annoyed by the delay and as a result may be unsatisfied with the ATM transaction. SUMMARY OF THE INVENTION It is among the objects of an embodiment of the present invention to obviate or mitigate one or more of the above disadvantages or other disadvantages associated with prior art self-service terminals. According to a first aspect of the present invention there is provided a self-service terminal having a user interface for interacting with a user, characterized in that the terminal includes sensing means for sensing physiological data associated with a user, analyzing means for analyzing the physiological data to deduce the user&#39;s emotional state, and control means responsive to the analyzing means for adapting the terminal&#39;s interaction with the user in response to the user&#39;s emotional state. The sensing means may be implemented using a contact device, but more preferably, using a non-contact device. Where a contact device is used, the sensing means may comprise a touch area incorporating sensors for determining the user&#39;s skin temperature, pulse rate, blood pressure, skin conductivity, and such like physiological data. A suitable touch area may be implemented by a device developed by IBM (trade mark) and called an “emotion mouse”. Where a non-contact device is used, and the terminal includes speech input, the sensing means may be implemented by a voice monitoring system for detecting changes in a user&#39;s voice. Additionally or alternatively, the sensing means may be implemented by facial recognition to detect changes in the user&#39;s facial appearance during a transaction. The sensing means may be implemented using an iris camera for imaging the user&#39;s iris and for detecting changes within the iris, such as changes to blood vessels, and such like. The sensing means may be implemented by a gesture recognition system. The analyzing means may be implemented by any convenient algorithm for deducing a person&#39;s emotional state from physiological measurements taken from the person. An overview of such algorithms is given in chapter 6 of “Affective Computing” by Rosalind W Picard, MIT Press, 1997, ISBN 0-262-16170-2. The control means may be implemented by a control application executing on the terminal. The control application may present a user with a sequence of screens to guide the user through a transaction. The control application may determine which screens are to be shown to the user in response to the user&#39;s emotional state as deduced by the analyzing algorithm. The term “screen” is used herein to denote the graphics, text, controls (such as menu options), and such like, that are displayed on an SST display; the term “screen” as used herein does not refer to the hardware (for example, the LCD, CRT, or touchscreen) that displays the graphics, text, controls, and such like. Typically, when a transaction is being entered at an SST, a series of screens are presented in succession on the SST display. For example, a first screen may request a user to insert a card, a second screen may invite the user to enter his/her personal identification number (PIN), a third screen may invite the user to select a transaction, and so on. By virtue of this aspect of the invention, the terminal is able to sense physiological data from a user during a transaction, analyze the data, and determine what to present to the user to comply with the user&#39;s emotional state. For example, the terminal may determine what transaction options to present, whether to present advertising or marketing material, if advertising is to be presented then what advertisements to present, for how long the advertisement is to last, at what point in the transaction the advertisement is to be shown, and such like. Thus, the user&#39;s experience at the SST can be improved by personalizing a transaction to the user&#39;s emotional state. For example, if a user feels insecure then the SST may: highlight to the user alternative SST locations at which they user may feel more secure; display a message regarding privacy and trust to reassure the user that the transaction is secure and that the transaction provider is one that the user can trust to keep user information private; give the user an option of more time to select a transaction option. The SST can improve the transaction provider&#39;s marketing and advertising efficiency by targeting advertisements that are known to be more effective for a particular emotional state. For example, if the user is in a happy mood and relaxed then the user might be receptive to advertising and the SST may: present humorous advertisements, and/or have a longer transaction sequence to provide more advertising time. The SST may record a user&#39;s emotional experience so that future transactions conducted by that user are automatically personalized. For example, if the user is not in a relaxed mood then the user might be irritated by advertising and the SST may: adapt the transaction to have no advertising during that transaction, or go to a customized quick transaction flow for that user the next time they use the SST. If the SST detects particular emotional states of users, then the SST may invoke extra security measures to improve security for both the users and the SST provider. For example, if the user was detected as being under a great deal of stress then this may indicate that the user is executing a transaction under duress, or the user may be using a stolen transaction token (such as a magnetic stripe card). These extreme types of emotional states could trigger additional security measures at the SST, such as: more security photographs being taken; and/or security information being requested from the user. In one embodiment the SST is an ATM. According to a second aspect of the present invention there is provided a method of operating a self-service terminal, the method comprising the steps of: sensing physiological data associated with a user of the terminal, analyzing the physiological data to deduce the user&#39;s emotional state, and adapting the terminal&#39;s interaction with the user in response to the user&#39;s emotional state. BRIEF DESCRIPTION OF THE DRAWINGS These and other aspects of the present invention will be apparent from the following specific description, given by way of example, with reference to the accompanying drawings, in which: FIG. 1 is a block diagram of a self-service terminal according to one embodiment of the present invention; FIG. 2 is a block diagram of a part (the controller) of the terminal of FIG. 1 ; FIGS. 3A to 3F illustrate a sequence of screens presented to one user of the terminal of FIG. 1 ; and FIGS. 4A to 4H illustrate a sequence of screens presented to another user of the terminal of FIG. 1. DETAILED DESCRIPTION Reference is now made to FIG. 1, which illustrates an SST 10 in the form of an ATM being operated by a user 12. The ATM 10 includes a user interface 14 for outputting information to a user and for allowing a user to input information. The ATM 10 also includes sensing means 16 in the form of a camera module 18 (that includes facial recognition software), a touch plate module 20 (implemented by an “emotion mouse”), and a microphone module 22 (that includes voice recognition software). The user interface 14 is a molded fascia incorporating: a display module 30, an encrypting keypad module 32, and a plurality of slots aligned with modules located behind the fascia. The slots include a card entry/exit slot (not shown) that aligns with a magnetic card reader/writer (MCRW) module 36, a printer slot (not shown) that aligns with a printer module 38, and a cash dispense slot (not shown) that aligns with a cash dispense module 40. The ATM 10 also includes an internal journal printer module 50 for creating a record of all transactions executed by the ATM 10, an ATM controller module 52 for controlling the operation of the various modules ( 18 to 50 ), and a network connection module 54 for communicating with a remote transaction host (not shown) for authorizing transactions. All of the modules ( 18 to 54 ) within the ATM 12 are interconnected by an internal bus 56 for securely conveying data. The ATM controller module 52 is shown in more detail in FIG. 2. The controller 52 comprises a BIOS 60 stored in non-volatile memory, a microprocessor 62, associated main memory 64, and storage space 66 in the form of a magnetic disk drive. In use, the ATM 12 loads an operating system kernel 70, an ATM application program 72, and a data analyzing program 74 into the main memory 64. The ATM application program 72 is used to control the operation of the ATM 12. In particular, the ATM application program 72 : provides the sequence of screens used in each transaction (referred to as the application flow); monitors the condition of each module within the ATM (state of health monitoring); and interfaces with the analyzing program 74. The analyzing program 74 implements a discriminant function analysis model for analyzing data received from the sensor modules 18 to 22 ; however, any other convenient analyzing program may be used. The analyzing program 74 processes data received from one or more of the sensor modules (camera 18, touch plate 20, or microphone 22 ) to deduce the emotional state of the user 12. The analyzing program 74 selects an emotion category that is the closest match to the user&#39;s emotional state, and outputs a code representing this category to the ATM application program 72. In this embodiment, the categories are: anger, hurriedness, fear, happiness, sadness, and surprise. The ATM application program 72 receives this code and adapts the transaction flow according to the emotional state represented by this code. This is implemented by the ATM application program 72 accessing a stored look-up table (not shown) having an index entry for each code. Each code in the look-up table has a unique transaction flow associated with it. An example of a typical transaction at the ATM 10 will now be described with reference to FIGS. 3A to 3F, which illustrate the sequence of screens presented to the user 12. When the user 12 approaches the ATM 10 he is presented with a welcome screen 80 a ( FIG. 3A ) on display 30 inviting him to insert his card. After inserting his card, the user 12 is presented with a screen 80 b ( FIG. 3B ) inviting him to enter his PIN, and the ATM application program 72 activates the sensors 18 to 22 to capture physiological data about the user 12. The ATM application receives data from the sensors 18 to 22 and conveys this data to the data analyzing program 74. Data analyzing program 74 processes the received data, deduces the user&#39;s emotional state from the data, generates a category code representing the user&#39;s emotional state, and conveys this code to the ATM application program 72. The ATM application program 72 accesses the look-up table (not shown) using the category code received from the data analyzing program 74 to determine what sequence of screens should be presented to the user 12. In this example, the user&#39;s state is hurriedness, so the sequence of screens is that for the shortest possible transaction time. The ATM application program 72 then presents the user 12 with a screen 80 c ( FIG. 3C ) listing transaction options available. After the user 12 has selected the withdraw cash option, the ATM application 72 presents the user with a screen 80 d ( FIG. 3D ) indicating cash amounts available. Once the user has selected a cash amount, the ATM application authorizes the transaction, presents a screen 80 e ( FIG. 3E ) inviting the user to remove his card, then a screen 80 f ( FIG. 3F ) inviting the user to remove the requested cash. An example of a typical transaction at the ATM 10 will now be described with reference to FIGS. 4A to 4H, which illustrate the sequence of screens presented to another user (or the same user as for FIGS. 3A to 3F but in a different emotional state). When the user approaches the ATM 10 he is presented with a welcome screen 82 a ( FIG. 4A ) on display 30 inviting him to insert his card. After inserting his card, the user is presented with a screen 82 b ( FIG. 4B ) inviting him to enter his PIN, and the ATM application program 72 activates the sensors 18 to 22 to capture physiological data about the user. As in the previous example, the ATM application 72 receives data from the sensors 18 to 22 and conveys this data to the data analyzing program 74. Data analyzing program 74 processes the received data, deduces the user&#39;s emotional state from the data, generates a category code representing the user&#39;s emotional state, and conveys this code to the ATM application program 72. The ATM application program 72 accesses its look-up table (not shown) using the category code received from the data analyzing program 74 to determine what sequence of screens should be presented to the user. In this example, the user&#39;s state is happiness, so the sequence of screens includes an advertisement for a holiday, and promotional material for a loan. The ATM application program 72 then presents the user with a screen 82 c ( FIG. 4C ) listing transaction options available. After the user has selected the withdraw cash option, the ATM application 72 presents the user with a screen 82 d ( FIG. 4D ) indicating cash amounts available. Once the user has selected a cash amount, the ATM application 72 authorizes the transaction, and presents the user with a screen 82 e ( FIG. 4E ) incorporating a video 84 (in MPEG format) advertising a holiday, the screen 82 e also includes text 86 informing the user that the requested transaction is being authorized. Once the video (which lasts approximately four seconds) has finished, the ATM application 72 then presents the user with a screen 82 f ( FIG. 4F ) incorporating promotional material 88 for a loan. The ATM application 72 then presents a screen 82 g ( FIG. 4G ) inviting the user to remove his card, and once the card has been removed, a screen 82 h ( FIG. 4H ) inviting the user to remove the requested cash. It will be appreciated that this embodiment has the advantage that a user is presented with a transaction sequence that is most likely to fulfil the user&#39;s expectations by matching a transaction to the user&#39;s emotional state. Various modifications may be made to the above described embodiment within the scope of the invention, for example, in other embodiments, the user may be asked to touch the touch plate 20 at the beginning of the transaction so that the touch plate can collect physiological data from the user&#39;s hand. In other embodiments, multiple algorithms may be used to implement the analyzing program 74, one for each sensor module 18 to 22. In other embodiments, different sensors may be used. In other embodiments, the touch plate sensor may be implemented on the keys of the encrypting keypad so that physiological measurements can be taken while the user is entering his PIN or other transaction details. In other embodiments, the user&#39;s emotional state may be continually monitored during the transaction flow so that the transaction flow may be changed at any point in response to the user&#39;s emotional state; for example, an advertisement may be stopped if a user&#39;s emotional state changes from being happy or relaxed to being unhappy or angry. In other embodiments, different emotional states may be categorized than those described in the above embodiment.

Summary: A self service terminal ( 10 ) and method of operating a self-service terminal ( 10 ) having a user interface ( 14 ) for interacting with a user ( 12 ). The terminal ( 10 ) includes a sensing device ( 18, 20, or 22 ) for sensing physiological data associated with a user, an analyzing device ( 74 ) for analyzing the physiological data to deduce the user&#39;s emotional state, and a controlling device ( 72 ) responsive to the analyzing device ( 74 ) for adapting the terminal&#39;s interaction with the user in response to the user&#39;s emotional state.

big_patent

en

Summarize: BACKGROUND 1. Field of the Invention The present invention relates to a contact lens insertion apparatus and, in particular, to an improved contact lens insertion apparatus that can be operated easily by anyone, even persons whose muscular control and coordination render it difficult or impossible to insert contact lenses. 2. Prior Art A variety of contact lens insertion devices utilizing a light source have been designed. Most of these have included a flash light type housing with a lens holder positioned at the top of the housing. A lens may be maintained in the holder in one of a variety of ways, including fluid surface tension, as described in U.S. Pat. No. 3,743,337 to Crary, or suction, as described in U.S. Pat. No. 3,304,113 to Hutchison. U.S. Pat. No. 3,139,298 to Grabiel teaches a device for inserting contact lenses wherein the lens is retained in a lens holder only by the force of gravity. Thus if the device is tipped to one side or the other the lens will fall off. U.S. Pat. Nos. 3,600,028 to Henning; 3,791,689 to Boone, et al.; 3,934,914 to Carruthers, and the above-identified patent to Hutchison all include means for creating a vacuum to hold a lens in position on a lens holder. Hutchison includes a bulb connected to the holder through pneumatic tubing. The bulb is squeezed and released in order to provide suction or pressure as needed to keep the lens in place or to release the lens. Boone, et al., and Carruthers teach a resiliant pouch surrounding a light path and communicating with a lens holder by way of a hole in the bottom of the lens holder. By squeezing the pouch, a partial vacuum may be created or released for operating the lens holder. Henning teaches another vacuum type arrangement wherein a tube can be pulled upward to create the partial vacuum and pushed downward to release the vacuum. U.S. Pat. No. 4,201,408 to Tressel teaches a similar method of creating a partial vacuum to hold the lens in place. An actuating member may be squeezed to evacuate air from a tube in communication with the lens holder. Tressel also teaches an end piece which can be placed against the cheek of the operator in order to steady the device. All of the patents utilizing a partial vacuum to hold the lens in position suffer from a common ailment. The lens holder includes a passageway leading to the bulb, pouch, etc. When the suction is created foreign matter may be, and frequently is, drawn into the passageway. This is an especially difficult area to clean, thus providing an environment for the growth of bacteria. In addition, when these devices release the lens, a small burst of air from the bulb, pouch, etc. thrusts some of this foreign matter onto the lens and into the eye of the operator. The net effect of the air passage leading to the lens holder renders these devices less than sanitary. U.S. Pat. No. 4,093,291 to Shurgin teaches a lens inserter that uses liquid adhesion to hold the lens to the inserter. Shurgin&#39;s device also contains a pair of tongs for grasping the side of the lens for removal thereof from the eye. While most of these designs use illumination to facilitate the proper positioning of the lens on the eye, various methods are used to accomplish the illumination. In particular, the light element may be contained in the housing, as in the Crary and Hutchison patents, or an external light source may be used whereby the illumination is conducted to the lens through either an optical fiber, as in the Boone, et al. patent, or a light pipe, as in the Carruthers patent. Both of these methods provide an illuminated target facilitating the guidance of the lens into the proper position on the eye. Some of these devices are easier to use than others. Some are in fact quite cumbersome. By way of example, none of the designs provides any means for opening the eyelid while steadying the device for persons who have difficulty holding either their hands or head steady. In addition, the means for providing suction to hold the lens in place frequently requires manipulation of a separate element, such as the squeeze bulb taught by Hutchison. This manipulation may be quite difficult for older people or those having less than full operation of their hands. SUMMARY OF THE INVENTION The present invention solves the problems that now exist by providing an apparatus for inserting contact lenses comprising a lens holder extension tube attached to a housing for providing a light channel; and a lens holder attached to the lens holder extension tube for holding a contact lens to be inserted by forming a partial vacuum between the lens and the lens holder, the lens holder including means at its center communicating with the light channel provided by the lens holder extension tube for allowing light to pass therethrough and thereby assisting the operator in guiding a lens. The invention also includes illuminating means contained within the housing and positioned at the end of the extension tube for providing illumination to the lens holder; a power source for powering the illuminating means; an eyelid opener extending from the housing for opening the lower eyelid of the eye, for steadying the applicator against the cheek bone of the user, and for guiding the lens into the proper position on the eye by pivoting the apparatus above the eyelid opener to bring the lens to the eye; a suction breaker operationally connected to the lens holder for allowing air to enter between the lens and the lens holder in order to break the suction of air therebetween and thereby easily remove the apparatus from the lens once the lens has been positioned on the eye; and a thumb holder attached to the housing in a position approximately 180° displaced from a suction breaker switch for facilitating the gripping of the apparatus by an individual and for facilitating the proper alignment of the individual&#39;s fingers upon the apparatus for easy operation thereof. Accordingly it is an object of the present invention to provide an improved contact lens insertion apparatus that is extremely easy to use. Another object of the present invention is to provide an improved contact lens insertion device that can be used, even in the dark, by people having extremely impaired eyesight. A further aim of the present invention is to provide an improved contact lens insertion device that can be used by people having an unsteady hand and/or head. It is an additional aspect of the present invention to provide an improved contact lens insertion device that contains a simple, yet reliable, suction and suction breaking mechanism for holding the lens in place on the applicator and for releasing the lens after contact has been made with the eye of the operator. A further object of the present invention is to provide an improved contact lens insertion device that maintains the lens in position by a partial vacuum, without containing an air passage leading to the lens holder, thusly avoiding the collection of unsanitary or harmful foreign matter within the device. Yet another aspect of the present invention is to provide an improved contact lens insertion device that contains an eyelid opener greatly facilitating the insertion of the contact lens onto the cornea of the eye. It is a further aim of the present invention to provide an improved contact lens insertion device that includes a thumb holder for facilitating the gripping of the applicator by the operator and insuring that the operator&#39;s hand is properly positioned to facilitate simple one-handed operation of the apparatus. Another object of the present invention is to provide an improved contact lens insertion device that is reliable, simply constructed, and inexpensive to manufacture. The foregoing objects, advantages, features and results of the present invention together with various other objects, advantages, features and results thereof which will be evident to those skilled in the art in light of this disclosure may be achieved with the exemplary embodiments of the invention described in detail hereinafter and illustrated in the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a side sectional view of the applicator showing the interrelationship of the various components. FIG. 2 is a partial plan cut away view showing the positioning of the suction release mechanism. FIG. 3 is a perspective view showing the insertion apparatus in use. DESCRIPTION OF THE PREFERRED EMBODIMENTS While the present invention is susceptible of various modifications and alternative constructions, an embodiment is shown in the drawings and will herein be described in detail. It should be understood however that it is not the intention to limit the invention to the particular forms disclosed; but, on the contrary, the intention is to cover all modifications, equivalences and alternative constructions falling within the spirit and scope of the invention as expressed in the appended claims. Users of contact lenses frequently encounter difficulty inserting the lens into position on their eye. Balancing the lens on a finger tip, the standard method of applying contact lenses, may prove impossible for an individual with an unsteady hand. Furthermore, the user may lack the requisite hand-eye coordination to properly place the lens on his eye with his finger. In addition, even normal eyes cannot focus on an object within one or two inches of the eye itself. Clearly the individual requiring corrective lenses frequently has greater difficulty in focusing on small and close objects than the person with normal vision. And, in special cases, like those involving people who have had the natural lens removed, in what is commonly called a cataract operation, vision is permanently and substantially impaired making it impossible to focus at all. Lens insertion devices usually attempt to solve the problems associated with inserting contact lenses by providing both a member to support the lens and a light to guide its insertion. The light is generally channeled from below the support member through the optical axis of the lens. This light is easily sighted as the lens is moved toward the eye. The channeled light provides a thin ray that can be seen when it is aligned coaxially with the optical axis of the eye. The correct position for an inserted lens is on the cornea of the eye with the optical axis of the lens coincident with the optical axis of the eye. By aligning the light with the optical axis of the eye the lens is automatically placed in the correct position for insertion. The present invention provides an improvement to the prior art contact lens insertion devices described hereinabove. Referring now to FIG. 1, a contact lens insertion apparatus 10 is shown including a housing 12, an extension tube 14 and a lens holder 16. The apparatus also includes an eyelid opener 18 which may be attached either to the housing or to a sleeve 20. The apparatus contains its own light source 22 to channel light through the extension tube 14. A light focusing tube 23 is positioned within the extension tube to narrow the beam of light to a pencil thin ray. The channeled light extends along the axis of the tube through the optical axis of a lens 24 shown in FIG. 3. This line of light greatly facilitates the alignment of the lens with the eye. In particular, it allows the user to be sure that he is inserting the lens to the proper position on his eye. The self contained light source is superior to the light pipes used in some of the prior art devices. The light pipes frequently required the operator to stare into a lamp. Thus, the channeled light was not easily differentiated from the background light. Although the optical fiber relieved the operator from staring at the lamp, it still required the availability of a light in the room. On the other hand, the present invention can be used in rooms with no light, or even in a darkened room. The self-contained light source may be a bulb 22, as shown in FIG. 1, or any other light generating means. The applicator includes a power source for the light. This avoids the inconvenience and difficulty associated with the power cord that otherwise would be necessary. The structure to achieve this is simple, reliable and inexpensive. The housing 12 contains the power source such as a battery 26 positioned in the usually suitable manner. The housing itself may be metal or high-impact plastic or any other suitable material strong enough to house the elements described herein and be handled by an operator in the manner shown in FIG. 3. The housing comprises a handle portion 28 and a neck portion 30. The handle portion contains a removable cap 32 at its end. The cap threadably engages the interior surface of the housing to detachably connect therewith. This detachable connection provides for easy access to replace the battery and light when necessary. A spring 34 provides an electrical connection between the anode of the battery and the light. The spring also provides a snug fit between the cathode of the battery and the removable cap 32. A light switch 36 comprises a button 38 and a resilient metal strip 40. Slight pressure on the button presses the metal strip against the cathode of the battery to complete an electrical connection with the light. The metal strip 40 is in contact with the lead 42 that contacts the negative terminal of the light in a manner to be described below. Release of the button 38 breaks the connection, since the strip 40 is biased to return to the non-contacting position. The metal strip 40 is so flexible that very little pressure is required to engage the strip with the cathode of the battery. Thus, by resting the little finger against the switch 36 the connection is completed. As previously mentioned the extension tube is hollow in order to form a light channel to direct the light to the lens. The lens holder, of course, also must have some provision to allow the thusly channelled light to pass through to the lens. Accordingly, the lens holder contains a hole 44 at its center. This hole is covered with a plastic window 46 in order to isolate the lens holder from the inside of the apparatus. An advantage of the lens holder is that it obviates the finger balancing act that must be performed by those not using any insertion device. Furthermore, some of the prior insertion devices rely upon gravity to hold the lens on top of a vertically positioned member. This type of device is not suitable for persons having difficulty controlling their head. A shaky head may knock the lens from the apparatus or knock the entire apparatus over. Also it may be difficult for such an individual to properly align his eye above the apparatus even though the light makes the individual fully aware of the location of the lens. As explained above, other insertion devices have used suction to maintain the lens in position on the holder, but the suction mechanism is generally quite cumbersome and/or complicated. Another advantage is that the lens does not have to be touched by an operator&#39;s fingers since pressing the holder against the convex surface of a lens securely positions the lens against the holder. Thus the lens may be picked up by the holder and washed or disinfected before insertion onto the eye without the lens ever coming into contact with a finger or hand. The lens holder achieves these and other advantages by providing a partially concave upper surface 48 and a lower surface 50. The upper surface conforms to the shape of a lens to form an air seal when a lens is pressed thereinto. A partial vacuum is created between the lens and the upper surface of the holder since pressing the lens into the holder forces air from between the two elements. This provides a suction that is sufficient to maintain the lens in position when the applicator is raised and turned to any position. The lens holder 16 (FIG. 1) must be constructed of flexible material that is soft enough to accept and hold a lens without damaging it. Rubber is a suitable material for this purpose. The hole 44, which is sealed with a transparent member such as the plastic window 46, allows the light from the light source 22 to pass through the center of the lens. In this way the suction is maintained while light is allowed to pass. In addition, there is no air passage between the lens holder and the interior of the apparatus. Thus the difficulties associated with the prior lens applicators arising from the entry of dirt and unsanitary foreign matter into the device are overcome. Another of the advantages of the present invention is the inclusion of a means for easily breaking the suction between the lens and lens holder after the lens has been placed on the eye. The combination of the suction and suction breaker provides an insertion device that can securely hold yet easily release the lens for positioning onto the eye. The suction breaking means herein described is very easily operated by one finger while the insertion device is gripped by the operator. This represents a marked improvement over previous insertion devices which either had no provision for releasing the lens or incorporated more complicated &#34;plumbing&#34; mechanisms which could not easily be manipulated by one finger of the hand holding the device. These devices also included the air passageway together with the disadvantages attendant therewith which were described above in detail. The suction release mechanism functions to partially pull the lens holder away from the lens in order to allow air to enter therebetween thereby breaking the suction. As seen in FIG. 2, a suction release switch 52 is connected via a string 54 to the lower surface 50 of the lens holder 16. Although a nylon string is preferable, the string properly may comprise any number of materials. For example, thin wire or strong thread may be employed successfully. A spring 56 is positioned between the lens holder and a spring stop 58. The spring should be positioned on one side of the lens holder, and displaced 90° from the suction release switch. This placement insures that the spring will not catch the eyelash of the operator. The spring 56 is positioned between the lens holder and a spring stop 58. The spring 54 runs through the center of the spring to connect with the lens holder. The spring biases the lens holder to return to its normal position and keep the string taut. A tube 60 extends between the spring stop 58 and a drip guard 62. The string is fed through this tube. As shown more clearly in FIG. 1, the string is attached to one end of the suction release switch 52. When the switch 52 is depressed the string is tightened which distends the shape of the lens holder 16 to break the vacuum and compress the spring. Upon release of the switch the spring 56 promptly returns the lens holder to its original position. Actuation of the suction release switch 52 also extinguishes the light 22. FIG. 1 illustrates a resilient metal strip 64 positioned beneath the switch 52 which biases the switch toward its open position and serves as a contact between the lamp and the lead 42. The location designated by the numeral 66 indicates the point of connection between the resilient metal strip and the lead. When the switch is depressed the metal strip is forced away from the lead 42 which opens the connection to turn off the lamp. Thus, depressing the switch simultaneously breaks the vacuum and extinguishes the lamp. The insertion operation thereby is simplified, since the extinguishment of the light serves as a signal to the operator to assure him that the suction between the lens and lens holder has been broken. A pair of screws 68 and 70 securely fix the extension tube 14 to the housing by friction fit. By loosening the screws the extension tube can be pulled slightly out of the housing or, contrariwise, pushed slightly further into the housing. This telescoping provides a means of adjusting the tension of the string. Should the string not be sufficiently taut, the extension tube can be moved outwardly from the housing, thereby tightening the string. Contrariwise, should the string be too tight, the extension tube can be moved slightly into the housing to reduce the tension in the string. The insertion apparatus is designed to be easily manipulated by anyone, including persons with severly impaired eyesight. The size of the apparatus together with the positioning of the various elements thereon facilitates the convenient operation of same with only one hand. The handle portion 28 of the housing 12 contains grooves 72 to facilitate gripping the apparatus. Manipulating the apparatus is further facilitated by a thumb holder 74, which has a concave shape in order to admit part of a thumb. The thumb holder is positioned on the housing opposite the suction release switch by a screw 76. This feature facilitates the grasping of the housing by the user and insures that the fingers of the user can easily operate the light switch 36 and the suction release switch 52. Thus by picking up the applicator and positioning the thumb in the thumb holder the user&#39;s fingers are automatically positioned adjacent to the operating switches of the apparatus, as seen in FIG. 3. A drip guard 62, seen best in FIG. 3, is attached to the extension tube 14. The guard encircles the extension tube 14 and abuts the inner wall of the sleeve 20 in order to prevent liquid from running onto the housing. Thus a lens securely held in place on the lens holder may be immersed in ophthalmic fluid, water, or other liquid for cleaning or disinfecting prior to insertion on the eye. The holder may be immediately rotated and yet the fluid will neither run over or into the housing nor flow onto the fingers of the operator. It is effectively blocked by the drip guard. The sleeve 20 can be slipped over the extension tube 14 to make a friction fit with the neck portion 30 of the housing 12. As mentioned, the drip guard 62 also tightly contacts the inside surface of the sleeve 20. Thus any accumulation of liquid will be contained within the sleeve and will not drip off onto the operator when the apparatus is tilted to invert the lens. Another advantage of the apparatus is that it includes means for steadying the device against the cheekbone of the operator and for holding the lower eyelid open during the insertion process. This means comprises the eyelid opener 18 which can be seen in FIGS. 1 and 2. The eyelid opener may be a U-shaped member swivally mounted to either the extension tube 14, the housing 12 or, as shown, the sleeve 20. The eyelid opener pulls the lower eyelid downwardly and also serves as a pivot upon which the applicator may be rotated to bring the lens closer to the eyeball as shown in FIG. 3. Once in place, the operator can look toward the device to see the light. When he sees the light, the contact lens is in the proper position to be inserted. The swivel mount enables the apparatus to be pushed directly toward the eye to insert the lens once it has been properly aligned with the eye. FIG. 1 illustrates that the eyelid opener can swivel up to 20° to facilitate the insertion operation by pivoting from the position shown in solid line to the position shown in dotted line. In operation a contact lens is pressed into the lens holder 14. This may be done without touching the lens. Pressing the lens into the holder evacuates the air between the lens and the holder and creates a vacuum therein. The vacuum holds the lens in place upon the holder. The user places his thumb within the thumb holder and grasps the housing as shown in phantom in FIG. 3. The natural placement of the fingers, the little finger at the light switch and the large finger at the release switch, allows the light switch 36 to be actuated thereby turning the light on. Since the switch is so sensitive, a slight touch with the little finger turns on the light. The eyelid opener is pressed immediately below the lower eyelid and moved slightly downwardly to open the eye. Using the eyelid opener as a pivot arm, the insertion device is swung up to place the lens close to the eye. The operator then looks toward the light and pushes the device straight towards the eye to place the lens onto his cornea. Once the lens is pressed against the eye the switch 52 is actuated to distend the lens holder 16. The distension allows air to enter between the lens and lens holder breaking the vacuum. At the same time as the release switch 52 breaks the vacuum, it also turns off the light to indicate to the operator that the suction has been broken. Thus the operator can then retract the apparatus.

Summary: A contact lens insertion apparatus incorporating an illuminated target for guiding the lens into the proper position on the cornea of an eye. The lens is held in place on the apparatus by a flexible cup that maintains a vacuum between itself and the lens. A suction breaking assembly enables easy removal of the lens from the apparatus once the lens has been properly positioned on the cornea. Also included is a lower eyelid opener which not only lowers the lid for easy insertion of the lens but also provides a pivot arm enabling the user to swing the applicator up, and easily and correctly position the lens on the eye.

big_patent

en

Summarize: By. Associated Press. and Daily Mail Reporter. PUBLISHED:. 18:49 EST, 12 January 2014. |. UPDATED:. 18:49 EST, 12 January 2014. A Georgia community has rallied around a Vietnam veteran who has been living out of his minivan with his dog for more than a year after being evicted from his home by mortgage giant Freddie Mac. John Chambers, 65, lost his house in Victory Park, Marietta, in January 2013 and has lived in a Walmart parking lot since, with his seven-year-old border collie Scout. Chambers says he was evicted without warning, lost his job after his employer learned he was homeless and has had trouble finding alternative housing through the Veterans Administration. Home: The minivan Chambers and his dog scout have called home for more than a year. Hope: Chambers was disheartened by the turn of events but has been touched by the outpouring of kindness from the community since his situtation was made public. Chambers is a former architectural draftsman who served in Vietnam and helped build facilities for U.S. troops in Iraq as a private contractor. The veteran's home is now owned by the Federal Home Loan Mortgage Corp., also known as Freddie Mac. Chambers sued the corporation, claiming that he was wrongfully evicted without proper notice and was led to believe he was being placed in a loan modification program. Freddie Mac spokesman, Brad German, says the corporation doesn't comment on pending litigation. Local paper the Marietta Daily Journal published a story about Chambers' situation earlier this week as he prepared to spend another night in his van while the region was hit with sub-freezing temperatures and wind chills. Since then, hundreds of people have offered money, temporary shelter and food for his dog, Scout, he said. 'I'm so overwhelmed I really don't know what to say,' Chambers told the newspaper before checking into an extended stay hotel room being paid for by the American Legion Post 29 in Marietta.'Tons of people have come out to the car, dropping off food, cards, asking how they can help, telling me I could stay the night in their house. This is surreal. I don't want to be in the limelight, really. I'm a little uncomfortable by it. But it humbles me. People coming by, giving me money, giving me groceries. People have been coming out of the woodwork all day,' he said. Cold world: College-educated architect and Vietnam War veteran John Chambers, who has been living homeless in his minivan since January of 2013, prepares for another cold night inside his mini-van in a local shopping center parking lot with his dog Scout. When he was evicted, Chambers approached a homeless shelter with Scout, but the facility did not accept dogs. So he chose to stay in his minivan with his beloved dog. The Veterans Administration was less than helpful.“They said they were going to try and find me some housing. I drove to East Point, and when I got there they said they didn’t have any housing,” he said. “I asked ‘Why did you make me drive all the way from Marietta to East Point when you could have told me that over the phone?’ And she said, ‘Oh, I didn’t want to but my supervisor told me to do it.’” Chambers told the Marietta Daily Journal.Chambers told the Marietta Daily Journal that at least 100 people have come to visit him in his minivan since the story was published, bringing cards, cash, dog food and offers of accommodation. Homeless: John Chambers checks on his former home in the Victory Park Community of Marietta, Georgia. His 186 Parkview Drive home was foreclosed on by Freddie Mac which he is currently fighting in court. The service officer for Post 29 of the American Legion, Ken Buechel, said the group's goal is to help Chambers find a new job and get back on his feet. Metro Atlanta real estate agent, Melody Unger, and her attorney have also formed a nonprofit group to try saving Chambers' home.'She's trying to get investors to actually buy my house, which would be my best bet because I'd like to drop this lawsuit like a hot potato,' Chamber said. He's spent more than $22,000 in legal fees trying to save his house, which is worth about $50,000.Unger said she contacted her attorney, Elizabeth Cook, and shared the story with her. The two women have now formed a nonprofit organization, JRC Home Fund Inc., to which people can donate money to help save Chambers’ home. Man's best friend: Chambers was unable to take Scout to a homeless shelter with him so chose to stay with the dog in his minivan. “If we can just get 100 people to donate $500 I feel like we could probably get the house for $50,000,” Unger said.Despite falling on hard times and facing uncertainty, Chambers said the outpouring of support has rekindled his optimism.'I was getting into a bitter state but it's a whole new world out there and I have a new life because of this,' he said. 'Just to have found one person in this world like all of the ones I've run into so far would have been enough to change my outlook. And here I've found more than a hundred.'

Summary: John Chamber, 65, has been living in his car with his dog Scout for more than a year. He says he was evicted from his Marietta, Georgia house by mortgage giant Freddie Mac without warning. The Vietnam vet lost his job after his employer learned he was homeless. After he spent a night in his car in record cold temperatures an attorney contacted a local paper about Chambers. The resulting article brought an outpouring of kindness from strangers who have been visiting his car bringing cash, dog food and offers of accommodation. Chambers says he was beginning to become negative and depressed by has been turned around by the kind people he has met. His attorney has begun a charity fund to help Chambers buy back his house.

cnn_dailymail

en

Summarize: So Yeon Ryu was seven years old when she gave her first violin recital. She was in love with music, but at age 12 she had to make a tough decision. "I started playing golf in elementary school. One day my golf coach took the team to a golf course and I fell in love with it. I loved walking the course and being out in nature," recalls Ryu, now 22. She started to notice several professional golfers, like fellow South Korean Grace Park. "She was fashionable, powerful and beautiful," Ryu says. Her golf was getting so much better that her mother asked what everyone had been thinking: "Do you want to become a violinist or a professional golfer?" The moment of truth. "It was one of the hardest decisions I have ever had to make. I always wanted to become violinist," Ryu acknowledges. By that time, she already had a role model: a girl who played on the elite U.S.-based LPGA Tour, and who, at age 20, became the first South Korean to claim a major title: Se Ri Pak. "She is a great trailblazer for all Korean golfers. She played super great on the LPGA and because of that we could dream about playing on the world stage," Ryu says. "But, this is not only her influence, we must be thankful to Grace Park, and Mi-Hyun Kim and other first generation Korean players on the LPGA." With her mind solely on golf, Ryu informed her family about her decision to put the violin to one side, though her mother was not convinced. "'Really? No violin?' My mom was so disappointed with my decision because everybody said I had a talent for music. I think my mom enjoyed choosing my recital dresses and having me perform on stage." The transition. Waking up early for practice was one of the toughest things in Ryu's new life. Little by little she got used to it and during the transition she discovered that her musical background would be very helpful. As a junior golfer, she had some trouble with her swing but her shots were good enough. "My swing tempo was consistently well and I think I got my good tempo from music," she says. In 2006, at age 16, Ryu represented South Korea in the Asian Games in Qatar and won gold in the individual and team events. Having impressed as an amateur, Ryu turned professional at the end of the following year and joined the KLPGA, the Korean women's tour. She won her first tournament as a KLPGA member, triumphing by four strokes at the Sports Seoul Open in April 2008. Her first winner's check brought her back to her first love -- she bought a violin for her sister, who had decided to pursue music as a career. The 2009 season would be even better for Ryu. Aged just 19, she won five times, earned over $500,000 and finished second in the Player of the Year race. But the real breakthrough came in 2011, at the most prestigious tournament in the female game: the U.S. Women's Open. After a shaky opening round that put her six strokes off the pace, Ryu shot 69 on Friday and Saturday to share the lead going into the final day. The tournament ended in a showdown between Ryu and her rival Hee Kyung Seo -- who had beaten her to the KLPGA player of the year award two years earlier. This time it was Ryu who prevailed. She forced a playoff with a birdie on the last regulation green before, after three extra holes, becoming just the fifth South Korean to win the major. A sixth, Na Yeon Choi, will defend her title at Sebonak Golf Club in Southampton, New York, this week. Ryu emulated her hero Pak -- who 13 years earlier became the first Korean to win it -- and, along with the winner's check of $585,000, it prompted her to make the next big step in her career. "I transferred from the KLPGA to the LPGA and people started to recognize me," she says. "I moved to the U.S. and I am now based in California. That was a pretty big change, relocating to another country." A keen student. Despite moving, Ryu continued studying physical education at Yonsei University, a private institution and the oldest of her country, and graduated in February this year with a bachelor's degree in sports management.. "I could not and would not trade my university life for anything. It was such a great time for me," she says. "Sometimes I couldn't sleep during a tournament because of assignments. I had to wake up early if I wanted to practice before going to class. Physically, it was a hard job but it was worth it." That work ethic goes some way to explaining the success of Korean women golfers and Ryu is now hoping that she can inspire new generations of young players like Pak and Kim did for her. "I'd love to be someone's role model. I want to share my experience and heart," says the golfer, who finished second behind compatriot and close friend Inbee Park at this season's opening major, the Kraft-Nabisco Championship in April,. "I would like to donate to poor people or junior golfers. I really want to support young golfers, not as an instructor but as a manager." The best decision. Ryu's determination at age 12 has led her to become one of the best players in women's golf; she currently sits fifth in the Rolex world rankings, just behind Choi, while Park is No. 1 after also winning this month's LPGA Championship. Last weekend the two friends battled for victory at the NW Arkansas Championship event, with Park beating Ryu in a playoff to claim her fifth win this season. "Inbee and I practice together a lot so when we are standing at the 18th hole, it feels like just a practice round," Ryu told reporters. "I wasn't really nervous. Two players cannot be champion, so she deserved it." Ryu was named 2012's LPGA Rookie of the Year, having won the 11th title of her pro career by a massive seven shots at the Jamie Farr Toldeo Classic. Her love for music is still alive, and Ryu acknowledges that it "is too hard" to even think of trading her accomplishments in golf for a night as concertmaster of the Berlin Philharmonic or Amsterdam's Royal Concertgebouw Orchestra. Her mother, who once was not satisfied with her daughter's decision, is now "so happy that I am a professional golfer." And it was from her mom that she received the advice that has shaped her career: "Enjoy your life."

Summary: South Korean golfer So Yeon Ryu has come a long way since her days as a musician. Ryu became the fifth player from her country to win the U.S. Women's Open in 2011. She will tee off at this week's 68th edition of the tournament in New York state. The 22-year-old was named the LPGA's Rookie of the Year for 2012.

cnn_dailymail

en