bluuebunny/crossref_metadata_2025 huggingface dataset:
Historical Deposition Influence in Residential Location Decisions: A Distance-Based GEV Model for Spatial Correlation
Authors: Chen Cynthia , Chen Jason , Timmermans Harry
Published: September 2009

Most of the existing literature on residential location choice is cross-sectional and assumes no history effect in people's residential location preferences. However, human beings are constantly adjusting themselves to the local environment and thus it seems reasonable to assume that preferences are formed over time. We hypothesize a historical deposition effect, which states that people's location preferences are likely to be a function of the attributes of where they lived before. Furthermore, this historical deposition influence can interact with lifecycle, such that it can become less important or insignificant during a life stage, like parenthood. Our study tests the existence of the historical deposition effect and analyzes its interaction with lifecycle. The study results support the historical deposition influence. Furthermore, we show that the historical deposition effect can be subdued by parenthood. The study also makes a methodological contribution by developing a distance-based GEV (generalized extreme value) model to account for spatial correlation. Spatial correlation is rarely treated in existing studies. The few existing studies typically assume a constant spatial correlation coefficient for adjacent zones. The distance-based GEV model relaxes the constant coefficient assumption and allows a distance-based correlation between nonadjacent zones. The results confirm this distance-based spatial correlation.
Where is Creativity in the City? Integrating Qualitative and GIS Methods
Authors: Brennan-Horley Chris , Gibson Chris
Published: September 2009

This paper discusses a new blend of methods developed to answer the question of where creativity is in the city. Experimentation with new methods was required because of empirical shortcomings with existing creative city research techniques; but also to respond to increasingly important questions of where nascent economic activities occur outside the formal sector, and governmental spheres of planning and economic development policy. In response we discuss here how qualitative methods can be used to address such concerns, based on experiences from an empirical project charged with the task of documenting creative activity in Darwin—a small city in Australia's tropical north. Diverse creative practitioners were interviewed about their interactions with the city—and hard-copy maps were used as anchoring devices around spatially orientated interview questions. Results from this interview-mapping process were accumulated and analysed in a geographical information system (GIS). Digital maps produced by this method revealed patterns of concentration and imagined ‘epicentres’ of creativity in Darwin, and showed how types of sites and spaces of the city are imagined as ‘creative’ in different ways. Qualitative mapping of creativity enabled the teasing out of contradictory and divergent stories of the location of creativity in the urban landscape. The opportunities which such methods present for researchers interested in how economic activities are ‘lived’ by workers, situated in social networks, and reproduced in everyday, material, spaces of the city are described.
An evergreen neotropical savanna tree (Gochnatia polymorpha, Asteraceae) produces different dry- and wet-season leaf types
Authors: Rossatto Davi R. , Kolb Rosana M.
Published: September 2009

Gochnatia polymorpha (Less.) Cabrera is a widespread tree species found in different physiognomies of neotropical savanna (cerrado) formations of south-eastern Brazil. The present study describes some leaf anatomical characteristics of this species as a function of the time of leaf flush, during dry or wet seasons. This species presents anatomical plasticity in the cuticle, palisade parenchyma and abaxial epidermis as well as in stomatal size and stomatal and trichome density, which are leaf structures linked with water-status control. Leaf structure changed to suit the particular environmental conditions during dry and wet seasons. The production of different wet- and dry-season leaf types in G. polymorpha could be a response to drought and an adaptation to environmental constraints in the cerrado.
Failure of the Sterile Air-Flow Component of a Protected Environment Detected by Demonstration of Chaetomium Species Colonization of Four Consecutive Immunosuppressed Occupants
Authors: Woods Gail L. , Davis J. Calvin , Vaughan William P.
Published: September 2009

Four bone marrow transplant recipients consecutively occupying the same room on our Oncology-Hematology Special Care Unit (OHSCU) became colonized with Chaetomium species between January and April, 1987. These patients, aged 27 to 43 years, were immunocompromised as a result of intensive chemotherapy, and were consequently at increased risk for development of invasive fungal infection. At the time of Chaetomium colonization, all patients were febrile, two had transient new infiltrates on chest x-ray, and three were receiving amphotericin B therapy. Subsequent environmental cultures revealed Chaetomium contamination of the OHSCU air-handling system, including the HEPA (high-efficiency particulate air) filters in seven of the nine rooms comprising the unit. Because fungal colonization of HEPA filters used to create a “protective environment” for immunocompromised patients can occur and can serve as a source for patient infections, guidelines concerning proper surveillance of these HEPA filters should be established. We suggest that before a new patient enters a “protected” room, the clean side of the HEPA filter should be cultured. If fungi are recovered from that culture, we would recommend changing the filter.
Enterococcal Infections: An Increasing Problem in Hospitalized Patients
Authors: Terpenning Margaret S. , Zervos Marcus J. , Schaberg Dennis R. , Kauffman Carol A.
Published: September 2009

We studied 157 episodes of infection or colonization with enterococci in 122 patients over a six-month period. One hundred twelve episodes (71.3%) occurred in patients over age 60 years. The most common sites for isolation of enterococci were the urinary tract, and bone and soft tissue. Nosocomial acquisition of enterococci occurred in 74.7% of all infections, and an additional 21% of episodes occurred in patients who had been transferred from another hospital or were regularly seen in the clinic. The overall mortality was 19.6%; 71.4% of those with bacteremia died. Enterococci appear to be significant pathogens, especially in older men in veterans' acute care hospitals and nursing home care units.
Diagnosis-Related Groups: Past and Future
Authors: McMahon Laurence F.
Published: September 2009

Introduction of the diagnosis-related group (DRG)-based Medicare Prospective Payment System is one of a series of major innovations that has occurred in the payment and delivery of health care over the past ten years. Changes such as the increased prevalence of health maintenance organizations, preferred provider organizations, third-party utilization review programs, and the peer review organizations for Medicare patients have all altered the way health care is financed and delivered. The DRG-based Medicare Prospective Payment System is the most visible of these changes, given its breadth of application and its radical departure from the previous retrospective reimbursement for hospital care. The Medicare Prospective Payment System has been in effect since October, 1983. As we approach the fifth anniversary of this program, it is a good time to review its history and to make some judgments as to its future.
Pseudomonas aeruginosa Infections Associated with Use of Povidone-Iodine in Patients Receiving Continuous Ambulatory Peritoneal Dialysis
Authors: Goetz Angella , Muder Robert R.
Published: September 2009

Fifteen episodes of infection due to Pseudomonas aeruginosa, including peritonitis and catheter site infections, occurred in nine patients receiving continuous ambulatory peritoneal dialysis over a 27-month period. Eight episodes were associated with catheter loss. Occurrence of P aeruginosa infection was significantly associated with use of povidone-iodine solution to cleanse the catheter site. There was no association with use of povidone-iodine solution to disinfect tubing connections, use of other skin care products or exposure to other environmental sources of P aeruginosa. Cultures of available povidone-iodine products were negative. Local irritation and alteration in skin flora caused by antiseptic solution or low-level contamination of povidone-iodine solution are potential mechanisms of infection.
Epidemic Bloodstream Infections Associated with Pressure Transducers: A Persistent Problem
Authors: Beck-Sague Consuelo M. , Jarvis William R.
Published: September 2009

Twenty-four outbreaks of nosocomial bloodstream infection (BSI) were investigated by the Centers for Disease Control from Jan 1, 1977 to Dec 31, 1987. Intravascular pressure monitoring devices (transducers) were the most commonly identified source of bacterial and fungal BSI outbreaks and were implicated as the source of infection in eight (33%) outbreaks. These included outbreaks caused by Candida parapsilosis (2), Serratia marcescens (2), Klebsiella oxytoca (1), Pseudomonas cepacia (1), Acinetobacter calcoaceticus (1), and one polymicrobial bacteremia outbreak due to Acinetobacter, Pseudomonas, Citrobacter, and Enterobacter species. In all eight outbreaks, reusable transducers improperly disinfected or fitted with domes that had been improperly sterilized served as reservoirs for the organism. Compared with nosocomial BSI outbreaks not related to transducers, those in which transducers were implicated as a reservoir involved a larger mean number of patients (24 v 9; P = 0.007), and were significantly more likely to involve intensive care unit patients (23/24 v 3/9; P = 0.025) and to have a longer mean duration (11 v 3 months; P= 0.007). These findings show that the characteristics of transducer- and non-transducer-related BSI outbreaks differ, and that centers using intravascular pressure monitoring devices must be aware of and implement recommended infection control strategies for care and maintenance of these devices.
Anaerobic Bacteremia in Patients with Acute Leukemia
Authors: Brown Elizabeth A. , Talbot George H. , Provencher Mary , Cassileth Peter
Published: September 2009

We reviewed 402 hospital admissions of patients with acute leukemia to define the frequency and characteristics of anaerobic bacteremia in this patient population. Six (5.2%) of the 116 septicemia episodes documented in these patients were caused by anaerobes (Bacteroides species, 3; Fusobacterium species, 2; and Clostridium tertium, 1); two of these episodes were polymicrobial. Five patients had had prior bacteremia. All six patients were receiving broad-spectrum antibiotics, including an anti-pseu-domonal penicillin, at the time of the episode. In each instance, the absolute granulocyte count was 0/mm3. Five patients had clinically apparent sources of infection, including perirectal abscess, gastrointestinal bleeding, or Clostridium difficile- associated diarrhea. Anaerobic bacteremia is an infrequent occurrence in granulocytopenic patients with acute leukemia, but may occur when there is obvious disruption of normal gastrointestinal anatomic barriers.
Occupational Exposure to Influenza-Introduction of an Index Case to a Hospital
Authors: Berlinberg Craig D. , Weingarten Scott R. , Bolton Linda B. , Waterman Steven H.
Published: September 2009

The epidemiology of influenza in the hospital is frequently confounded by failure to separate community-acquired from nosocomial transmission. An 83-year-old woman was hospitalized one day after returning from Asia with complications resulting from acute influenza A (H3N2) infection; she was the first culture-confirmed case in the region during the 1987-1988 influenza season, and her illness antedated other influenza cases in the area by at least four weeks. The patient shed virus at least four days after admission and transmitted influenza to her primary physician; both had received trivalent influenza vaccine four weeks earlier. Surveillance data from the 28 health care providers (HCPs) in contact with the index case (mean age: 34.5 years; median time of contact: four hours, none receiving vaccine) revealed no evidence of transmission as detected by paired type-specific complement-fixation antibodies and throat culture (20 subjects) or acute serologies and culture (7 subjects). No febrile respiratory illnesses were detected among other patients on the same ward, although three were reported among HCPs. Thus, neither secondary spread of influenza from infected patient to hospital HCPs nor nosocomial transmission apparently took place, although transmission did occur to the primary physician.
Evaluation of New In Vitro Diagnostic Test Procedures in Clinical Microbiology
Authors: Washington John A.
Published: September 2009

In vitro diagnostic products (IVDs) for infectious diseases are becoming available commercially at an ever-increasing rate. Aggressive marketing of such products often creates pressure on the laboratory to use IVDs, sometimes even before independent evaluations can be published in peer-reviewed scientific journals.

IVDs potentially offer simplicity and speed, particularly in detecting microorganisms that have fastidious growth requirements or that are simply not cultivable with currently available methodology. In some instances, IVDs may obviate the need for culture of more common microorganisms and may thereby allow testing to be done in physicians' offices and perhaps ultimately in the home.
A No-Rinse Alcohol Antiseptic and a No-Touch Dispenser for Hand Decontamination
Authors: Cronin Wendy A. , Gröschel Dieter H.M
Published: September 2009

Handwashing has longbeen recognized as the main way to prevent the spread of infectious agents. For routine handwashing in hospitals and clinics, antimicrobial agents are probably not needed. However, in certain high-risk situations it makes sense to use an antimicrobial agent to remove transient and reduce resident microorganisms.

Alcohol has been described as the “gold standard” for surgical skin preparations it will kill gram-positive and gram-negative bacteria, Myt obacterium tuberculosis, fungi, and many viruses. Alcohol is colorless, has its own cleaning action, and evaporates quickly from the skin.
Characterization of the herpes simplex virus (HSV)-1 tegument protein VP1-2 during infection with the HSV temperature-sensitive mutant tsB7
Authors: Abaitua F. , Souto R. N. , Browne H. , Daikoku T. , O'Hare P.
Published: September 2009

VP1-2, encoded by the UL36 gene of herpes simplex virus (HSV), is a large structural protein, conserved across the family Herpesviridae, that is assembled into the tegument and is essential for virus replication. Current evidence indicates that VP1-2 is a central component in the tegumentation and envelopment processes and that it also possesses important roles in capsid transport and entry. However, any detailed mechanistic understanding of VP1-2 function(s) remains limited. This study characterized the replication of HSV-1 tsB7, a temperature-sensitive mutant restricted at the non-permissive temperature due to a defect in VP1-2 function. A tsB7 virus expressing green fluorescent protein-fused VP16 protein was used to track the accumulation and location of a major tegument protein. After infection at the permissive temperature and shift to the non-permissive temperature, the production of infectious virus ceased. VP1-2 accumulated in altered cytosolic clusters, together with VP16 and other virion proteins. Furthermore, correlating with the results of immunofluorescence, electron microscopy demonstrated abnormal cytosolic capsid clustering and a block in envelopment. As VP1-2 encompasses a ubiquitin-specific protease domain, the occurrence of ubiquitin-conjugated proteins during tsB7 infection was also examined at the non-permissive temperature. A striking overaccumulation was observed of ubiquitin-specific conjugates in cytoplasmic clusters, overlapping and adjacent to the VP1-2 clusters. These results are discussed in relation to the possible functions of VP1-2 in the assembly pathway and the nature of the defect in tsB7.
Novel begomovirus species of recombinant nature in sweet potato (Ipomoea batatas) and Ipomoea indica: taxonomic and phylogenetic implications
Authors: Lozano Gloria , Trenado Helena P. , Valverde Rodrigo A. , Navas-Castillo Jesús
Published: September 2009

Viral diseases occur wherever sweet potato (Ipomoea batatas) is cultivated and because this crop is vegetatively propagated, accumulation and perpetuation of viruses can become a major constraint for production. Up to 90 % reductions in yield have been reported in association with viral infections. About 20 officially accepted or tentative virus species have been found in sweet potato and other Ipomoea species. They include three species of begomoviruses (genus Begomovirus, family Geminiviridae) whose genomes have been fully sequenced. In this investigation, we conducted a search for begomoviruses infecting sweet potato and Ipomoea indica in Spain and characterized the complete genome of 15 isolates. In addition to sweet potato leaf curl virus (SPLCV) and Ipomoea yellowing vein virus, we identified three new begomovirus species and a novel strain of SPLCV. Our analysis also demonstrated that extensive recombination events have shaped the populations of Ipomoea-infecting begomoviruses in Spain. The increased complexity of the unique Ipomoea-infecting begomovirus group, highlighted by our results, open new horizons to understand the phylogeny and evolution of the family Geminiviridae.
Sorting signals in the measles virus wild-type glycoproteins differently influence virus spread in polarized epithelia and lymphocytes
Authors: Runkler Nicole , Dietzel Erik , Carsillo Mary , Niewiesk Stefan , Maisner Andrea
Published: September 2009

The spread of virus infection within an organism is partially dictated by the receptor usage of the virus and can be influenced by sorting signals present in the viral glycoproteins expressed in infected cells. In previous studies, we have shown that the haemagglutinin (H) and fusion protein (F) of the measles virus (MV) vaccine strain MVEdm harbour tyrosine-dependent sorting signals which influence virus spread in both lymphocytes and epithelial cells to a similar degree. In contrast with the vaccine strain, MV wild-type virus does not use CD46 but CD150/SLAM and a not clearly identified molecule on epithelial cells as receptors. To determine differences in viral spread between vaccine and wild-type virus, we generated recombinant MV expressing glycoproteins of both the wild-type strain WTFb and the corresponding tyrosine mutants. In contrast with observations based on vaccine virus glycoproteins, mutations in wild-type virus H and F differently influenced cell-to-cell fusion and replication in polarized epithelia and lymphocytes. For wild-type H, our data suggest a key role of the cytoplasmic tyrosine signal for virus dissemination in vivo. It seems to be important for efficient virus spread between lymphocytes, while the tyrosine signal in the F protein gains importance in epithelial cells as both signals have to be intact to allow efficient spread of infection within epithelia.
A novel plant virus with unique properties infecting Japanese holly fern
Authors: Valverde Rodrigo A. , Sabanadzovic Sead
Published: September 2009

A novel RNA virus with a bipartite genome has been found associated with an emerging disease affecting Japanese holly fern (Cyrtomium falcatum). Diseased Japanese holly fern plants showed a variety of foliar symptoms and reduction in size. The virus was transmitted by grafting, as well as through spores from an infected plant. Partially purified preparations of the virus from infected ferns contained quasi-spherical particles that ranged from 30 to 40 nm in diameter. Double-stranded RNA (dsRNA) analyses from diseased plants yielded two major molecules of approximately 6.2 and 3.0 kbp in size, together with three other dsRNAs ascertained to be the replicative forms of subgenomic RNAs. The organization of RNA1 of this novel virus resembles that of raspberry bushy dwarf virus (genus Idaeovirus), whereas the genomic RNA2 showed a distinct organization and evolutionary origin. Results of this study indicate that the virus detected in diseased ferns is an undescribed phytovirus, for which the name Japanese holly fern mottle virus (JHFMoV) is proposed. Furthermore, we postulate that JHFMoV has enough distinguishing features to represent the type species of a novel genus of plant viruses. Taking into account the original host of the virus, we propose the name Pteridovirus for this taxon.
Molecular characterization of the plant virus genus Ourmiavirus and evidence of inter-kingdom reassortment of viral genome segments as its possible route of origin
Authors: Rastgou M. , Habibi M. K. , Izadpanah K. , Masenga V. , Milne R. G. , Wolf Y. I. , Koonin E. V. , Turina M.
Published: September 2009

Ourmia melon virus (OuMV), Epirus cherry virus (EpCV) and Cassava virus C (CsVC) are three species placed in the genus Ourmiavirus. We cloned and sequenced their RNA genomes. The sizes of the three genomic RNAs of OuMV, the type member of the genus, were 2814, 1064 and 974 nt and each had one open reading frame. RNA1 potentially encoded a 97.5 kDa protein carrying the GDD motif typical of RNA-dependent RNA polymerases (RdRps). The putative RdRps of ourmiaviruses are distantly related to known viral RdRps, with the closest similarity and phylogenetic affinity observed with fungal viruses of the genus Narnaviridae. RNA2 encoded a 31.6 kDa protein which, expressed in bacteria as a His-tag fusion protein and in plants through agroinfiltration, reacted specifically with antibodies made against tubular structures found in the cytoplasm. The ORF2 product is significantly similar to movement proteins of the genus Tombusviridae, and phylogenetic analysis supported this evolutionary relationship. The product of OuMV ORF3 is a 23.8 kDa protein. This protein was also expressed in bacteria and plants, and reacted specifically with antisera against the OuMV coat protein. The sequence of the ORF3 protein showed limited but significant similarity to capsid proteins of several plant and animal viruses, although phylogenetic analysis failed to reveal its most likely origin. Taken together, these results indicate that ourmiaviruses comprise a unique group of plant viruses that might have evolved by reassortment of genomic segments of RNA viruses infecting hosts belonging to different eukaryotic kingdoms, in particular, fungi and plants.
High-throughput sequence analysis of variants of human cytomegalovirus strains Towne and AD169
Authors: Bradley Amanda J. , Lurain Nell S. , Ghazal Peter , Trivedi Urmi , Cunningham Charles , Baluchova Katarina , Gatherer Derek , Wilkinson Gavin W. G. , Dargan Derrick J. , Davison Andrew J.
Published: September 2009

The genomes of commonly used variants of human cytomegalovirus (HCMV) strains Towne and AD169 each contain a substantial mutation in which a region (UL/b′) at the right end of the long unique region has been replaced by an inverted duplication of a region from the left end of the genome. Using high-throughput technology, we have sequenced HCMV strain Towne (ATCC VR-977) and confirmed the presence of two variants, one exhibiting the replacement in UL/b′ and the other intact in this region. Both variants are mutated in genes RL13, UL1, UL40, UL130, US1 and US9. We have also sequenced a novel AD169 variant (varUC) that is intact in UL/b′ except for a small deletion that affects genes UL144, UL142, UL141 and UL140. Like other AD169 variants, varUC is mutated in genes RL5A, RL13, UL36 and UL131A. A subpopulation of varUC contains an additional deletion affecting genes IRS1, US1 and US2.
Assembly of monomeric human cytomegalovirus pUL104 into portal structures
Authors: Holzenburg Andreas , Dittmer Alexandra , Bogner Elke
Published: September 2009

In order for human cytomegalovirus (HCMV) to replicate, concatemeric DNA has to be cleaved into unit-length genomes and packaged into preformed capsids. For packaging to take place and DNA to be translocated, a channel is required in the capsid. Viral capsid channels are generally formed by portal proteins. Here, we show by cross-linking, native gel electrophoresis of infected cells and gel permeation chromatography that the HCMV portal candidate protein pUL104 can form dimers and higher order multimers. Electron microscopy of purified monomeric pUL104 after 5 min incubation revealed that the protein had assembled into a multimeric form and that this form closely resembles complete portal assembly. This is the first study to show that pUL104 monomers have the ability to form portal complexes without additional viral proteins.
Functional analysis of the Bunyamwera orthobunyavirus Gc glycoprotein
Authors: Shi Xiaohong , Goli Josthna , Clark Gordon , Brauburger Kristina , Elliott Richard M.
Published: September 2009

The virion glycoproteins Gn and Gc of Bunyamwera orthobunyavirus (family Bunyaviridae) are encoded by the M RNA genome segment and have roles in both viral attachment and membrane fusion. To investigate further the structure and function of the Gc protein in viral replication, we generated 12 mutants that contain truncations from the N terminus. The effects of these deletions were analysed with regard to Golgi targeting, low pH-dependent membrane fusion, infectious virus-like particle (VLP) formation and virus infectivity. Our results show that the N-terminal half (453 residues) of the Gc ectodomain (909 residues in total) is dispensable for Golgi trafficking and cell fusion. However, deletions in this region resulted in a significant reduction in VLP formation. Four mutant viruses that contained N-terminal deletions in their Gc proteins were rescued, and found to be attenuated to different degrees in BHK-21 cells. Taken together, our data indicate that the N-terminal half of the Gc ectodomain is dispensable for replication in cell culture, whereas the C-terminal half is required to mediate cell fusion. A model for the domain structure of the Gc ectodomain is proposed.
Genetic evidence of two stocks of the whitemouth croaker Micropogonias furnieri in the Río de la Plata and oceanic front in Uruguay
Authors: Pereira A. N. , Márquez A. , Marin M. , Marin Y.
Published: September 2009

Analysis of the mitochondrial DNA control region resolved two stocks of whitemouth croaker Micropogonias furnieri in Uruguayan waters, one in the Río de la Plata and the other on the Uruguayan shelf. The whitemouth croaker is the most important coastal fishery resource along the coast and has the greatest commercial importance in the Río de la Plata and its oceanic front. The number of pair‐wise differences (π) and haplotype diversity (h) showed significant differences between the two regions. Frequencies of mtDNA haplotypes did not differ between Río de la Plata from Bahia Blanca M. furnieri. Samples from the oceanic front showed greater genetic variability and a larger effective number of females that were an order of magnitude larger than that in Río de la Plata. Mismatch distributions showed evidence of a recent population expansion in the oceanic region, beginning c. 40 000 b.p. The presence of two stocks of the M. furnieri in the study area should be considered in the management of this species' fishery.
An unusual ZZ/ZW sex chromosome system inCharacidiumfishes (Crenuchidae, Characiformes) with the presence of rDNA sites
Authors: Noleto R. B. , Amorim A. P. , Vicari M. R. , Artoni R. F. , Cestari M. M.
Published: September 2009

Characidiumfishes with a sex chromosome system form a monophyletic group. This work presents data ofCharacidium laneifrom the South Atlantic basin (Brazil), including an unknown type of ZW sex chromosome system for the groups including the presence of rDNA sites on sex chromosomes.
Phase I study of TLR9 agonist PF‐3512676 in combination with carboplatin and paclitaxel in patients with advanced non‐small‐cell lung cancer
Authors: Yamada Kazuhiko , Nakao Masao , Fukuyama Chikara , Nokihara Hiroshi , Yamamoto Noboru , Sekine Ikuo , Kunitoh Hideo , Ohe Yuichiro , Ohki Emiko , Hashimoto Junichi , Tamura Tomohide
Published: September 2009

This phase I, open‐label study investigated the Toll‐like receptor 9 agonist, PF‐3512676, in combination with carboplatin and paclitaxel in Japanese patients with advanced, non‐small‐cell lung cancer (NSCLC). Patients (n = 12) with treatment‐naive stage IIIB or IV NSCLC received single‐agent PF‐3512676 subcutaneously once during the first 7 days (monotherapy phase) in three escalating dose levels (0.1, 0.2, and 0.4 mg/kg) followed by a combination phase during which patients received 0.1 or 0.2 mg/kg PF‐3512676 subcutaneously on days 8 and 15 of each 3‐week cycle of carboplatin (area under the curve, 6 mg × min/mL) and paclitaxel (200 mg/m2). Safety and pharmacokinetics of PF‐3512676 were assessed during monotherapy and combination therapy phases. PF‐3512676 was tolerable as monotherapy or in combination with chemotherapy in patients with NSCLC. Most common treatment‐related, non‐hematologic adverse events (AEs) throughout the study were injection‐site reactions (n = 12, 100%) and flu‐like symptoms (n = 11, 91.7%) that were each grade 1 or 2 in all but one patient. All patients experienced neutropenia and leukopenia (≥grade 3 in 11 [91.7%] and seven [58.3%] patients, respectively). One patient in dose level 2 had a dose‐limiting toxicity: grade 3 rash and grade 3 increase in γ‐glutamyltransferase during combination therapy. Mean PF‐3512676 half‐life ranged from 4.8 to 21.6 h (longer with higher doses). Four (33%) patients had objective responses (one complete response, three partial responses), and seven (58%) patients achieved stable disease. PF‐3512676 as monotherapy and in combination with chemotherapy had an acceptable safety profile in Japanese patients with treatment‐naive NSCLC. (Cancer Sci 2009)
Vitamin E succinate induced apoptosis and enhanced chemosensitivity to paclitaxel in human bladder cancer cells in vitro and in vivo
Authors: Kanai Kunimitsu , Kikuchi Eiji , Mikami Shuji , Suzuki Eriko , Uchida Yasumitsu , Kodaira Kiichiro , Miyajima Akira , Ohigashi Takashi , Nakashima Jun , Oya Mototsugu
Published: September 2009

There have been several studies on the antitumor activities of vitamin E succinate (α‐TOS) as complementary and alternative medicine. In the present study, we investigated the cytotoxic effect of α‐TOS and the enhancement of chemosensitivity to paclitaxel by α‐TOS in bladder cancer. KU‐19‐19 and 5637 bladder cancer cell lines were cultured in α‐TOS and/or paclitaxel in vitro. Cell viability, flow cytometric analysis, and nuclear factor‐kappa B (NF‐κB) activity were analyzed. For in vivo therapeutic experiments, pre‐established KU‐19‐19 tumors were treated with α‐TOS and/or paclitaxel. In KU‐19‐19 and 5637 cells, the combination treatment resulted in a significantly higher level of growth inhibition, and apoptosis was significantly induced by the combination treatment. NF‐κB was activated by paclitaxel; however, the activation of NF‐κB was inhibited by α‐TOS. Also, the combination treatment significantly inhibited tumor growth in mice. In the immunostaining of the tumors, apoptosis was induced and proliferation was inhibited by the combination treatment. Combination treatment of α‐TOS and paclitaxel showed promising anticancer effects in terms of inhibiting bladder cancer cell growth and viability in vitro and in vivo. One of the potential mechanisms by which the combination therapy has synergistic cytotoxic effects against bladder cancer may be that α‐TOS inhibits NF‐κB induced by chemotherapeutic agents. (Cancer Sci 2009;)
Effects of Src inhibitors on cell growth and epidermal growth factor receptor and MET signaling in gefitinib‐resistant non‐small cell lung cancer cells with acquired MET amplification
Authors: Yoshida Takeshi , Okamoto Isamu , Okamoto Wataru , Hatashita Erina , Yamada Yuki , Kuwata Kiyoko , Nishio Kazuto , Fukuoka Masahiro , Jänne Pasi A. , Nakagawa Kazuhiko
Published: September 2009

The efficacy of epidermal growth factor receptor (EGFR)–tyrosine kinase inhibitors such as gefitinib and erlotinib in non‐small cell lung cancer (NSCLC) is often limited by the emergence of drug resistance conferred either by a secondary T790M mutation of EGFR or by acquired amplification of the MET gene. We now show that the extent of activation of the tyrosine kinase Src is markedly increased in gefitinib‐resistant NSCLC (HCC827 GR) cells with MET amplification compared with that in the gefitinib‐sensitive parental (HCC827) cells. In contrast, the extent of Src activation did not differ between gefitinib‐resistant NSCLC (PC9/ZD) cells harboring the T790M mutation of EGFR and the corresponding gefitinib‐sensitive parental (PC9) cells. This activation of Src in HCC827 GR cells was largely abolished by the MET‐TKI PHA‐665752 but was only partially inhibited by gefitinib, suggesting that Src activation is more dependent on MET signaling than on EGFR signaling in gefitinib‐resistant NSCLC cells with MET amplification. Src inhibitors blocked Akt and Erk signaling pathways, resulting in both suppression of cell growth and induction of apoptosis, in HCC827 GR cells as effectively as did the combination of gefitinib and PHA‐665752. Furthermore, Src inhibitor dasatinib inhibited tumor growth in HCC827 GR xenografts to a significantly greater extent than did treatment with gefitinib alone. These results provide a rationale for clinical targeting of Src in gefitinib‐resistant NSCLC with MET amplification. (Cancer Sci 2009)
Contemporary and historical impacts of megaherbivores on the population structure of tree euphorbias in South African subtropical thicket
Authors: Cowling Richard M. , Kamineth Abigail , Difford Mark , Campbell Eileen E.
Published: September 2009

African elephant and black rhinoceros – both megaherbivores – impact negatively on the abundance of succulent plants, including tree succulents, in South Africa’s subtropical thicket. We sampled 35 sites in subtropical thicket to assess historical and contemporary impacts of African elephant and black rhinoceros on the population structure of three species of succulent tree euphorbia. Population age structures were highly variable and showed no species‐specific effects. Almost half the sites had growing populations dominated by young individuals. Sites having contemporary impacts had significantly fewer individuals in the 30–75 years age range, but this effect declined with increasing terrain slope. Eighty‐one percent of sites assumed to have been impacted by megaherbivores historically had individuals that predated rhino and elephant extirpation in the region. This and other population age structure data indicate that megaherbivores and tree euphorbias coexisted historically on terrain of low relief. However, in areas not subject to contemporary impacts, euphorbia populations are likely to be much higher now than historically, owing to population relaxation after the extirpation of megaherbivores in the mid 1800s. Given the sensitivity of tree euphorbias to megaherbivore impacts, managers should consider monitoring the population structure of these species in order to identify thresholds of potential concern regarding megaherbivore impacts.
Bi‐directional activation between mesenchymal stem cells and CLL B‐cells: implication for CLL disease progression
Authors: Ding Wei , Nowakowski Grzegorz S. , Knox Traci R. , Boysen Justin C. , Maas Mary L. , Schwager Susan M. , Wu Wenting , Wellik Linda E. , Dietz Allan B. , Ghosh Asish K. , Secreto Charla R. , Medina Kay L. , Shanafelt Tait D. , Zent Clive S. , Call Timothy G. , Kay Neil E.
Published: September 2009

It was hypothesized that contact between chronic lymphocytic leukaemia (CLL) B‐cells and marrow stromal cells impact both cell types. To test this hypothesis, we utilized a long‐term primary culture system from bone biopsies that reliably generates a mesenchymal stem cell (MSC). Co‐culture of MSC with CLL B‐cells protected the latter from both spontaneous apoptosis and drug‐induced apoptosis. The CD38 expression in previously CD38 positive CLL B‐cells was up‐regulated with MSC co‐culture. Upregulation of CD71, CD25, CD69 and CD70 in CLL B‐cells was found in the co‐culture. CD71 upregulation was more significantly associated with high‐risk CLL, implicating CD71 regulation in the microenvironment predicting disease progression. In MSC, rapid ERK and AKT phosphorylation (within 30 min) were detected when CLL B‐cells and MSC were separated by transwell; indicating that activation of MSC was mediated by soluble factors. These findings support a bi‐directional activation between bone marrow stromal cells and CLL B‐cells.
Suppression of the hepcidin‐encoding gene Hamp permits iron overload in mice lacking both hemojuvelin and matriptase‐2/TMPRSS6
Authors: Truksa Jaroslav , Gelbart Terri , Peng Hongfan , Beutler Ernest , Beutler Bruce , Lee Pauline
Published: September 2009

Hepcidin, the master regulator of enteric iron absorption, is controlled by the opposing effects of pathways activated in response to iron excess or iron attenuation. Iron excess is regulated through a pathway involving the cell surface receptor hemojuvelin (HFE2) that stimulates expression of the hepcidin encoding gene (HAMP). Iron attenuation is countered through a pathway involving the hepatocyte‐specific plasma membrane protease matriptase‐2 encoded by TMPRSS6, leading to suppression of HAMP expression. The non‐redundant function of hemojuvelin and matriptase‐2 has been deduced from the phenotype imparted by mutations of HFE2 and TMPRSS6, which cause iron excess and iron deficiency respectively. Hemojuvelin is positioned to be the ideal substrate for matriptase‐2. To examine the relationship between hemojuvelin and matriptase‐2 in vivo, we crossed mice lacking the protease domain of matriptase‐2 with mice lacking hemojuvelin. Mice lacking functional matriptase‐2 and hemojuvelin exhibited low Hamp (Hamp1) expression, high serum and liver iron, and high transferrin saturation. Surprisingly, the double mutant mice also exhibited lower levels of iron in the heart compared to hemojuvelin‐deficient mice, demonstrating a possible cardioprotective effect resulting from the loss of matriptase‐2. This phenotype is consistent with hemojuvelin being a major substrate for matriptase‐2/TMPRSS6 protease activity.
Identification of pyrethroid resistance associated mutations in the para sodium channel of the two‐spotted spider mite Tetranychus urticae (Acari: Tetranychidae)
Authors: Tsagkarakou A. , Van Leeuwen T. , Khajehali J. , Ilias A. , Grispou M. , Williamson M. S. , Tirry L. , Vontas J.
Published: September 2009

We investigated pyrethroid resistance mechanisms in Tetranychus urticae strains from Greece. Combined bioassay, biochemical and synergistic data indicated that although P450 mono‐oxygenase activities were associated with the trait, target site insensitivity was the major resistance component. A 3.3 kb cDNA fragment of the T. urticae para sodium channel gene encompassing segment 4 of domain II to segment 6 of domain IV was obtained by a degenerate PCR strategy. The T. urticae sequence showed highest identity (56%) to the scabies mite, Sarcoptes scabiei, and was phylogenetically classified within the divergent group of Arachnida. Comparison of resistant and susceptible strains identified the point mutation F1538I in segment 6 of domain III, which is known to confer strong resistance to pyrethroids, along with a second mutation (A1215D) in the intracellular linker connecting domains II and III with an unknown role. Three transcripts were identified corresponding to the k and l alternative exons. The mode of inheritance of resistance was confirmed as incompletely recessive, which is consistent with a target site mechanism for pyrethroids.
Identification of a cis‐regulatory element required for 20‐hydroxyecdysone enhancement of antimicrobial peptide gene expression in Drosophila melanogaster
Authors: Zhang Z. , Palli S. R.
Published: September 2009

The antimicrobial peptide diptericin plays an important role in defence against microorganisms. Drosophila melanogaster diptericin mRNA levels showed an increase during the late final instar larval stage when the ecdysteroid titres increase to initiate metamorphosis. Deletion analysis in Drosophila melanogaster mbn2 (mbn2) cells identified a cis‐regulatory element (AAGAAAGATCCCCTG) necessary for 20‐hydroxyecdysone enhancement of peptidoglycan‐induced expression of diptericin in the 3 kb diptericin promoter. Proteins extracted from mbn2 cells treated with peptidoglycan plus 20‐hydroxyecdysone specifically bound to this element. 20‐hydroxyecdysone also enhanced peptidoglycan‐induced expression of four other antimicrobial peptide (AMP) genes –drosomycin, attacin‐A, metchnikowin and cecropin A1. Moreover, in silico promoter analysis using the meme program showed that an eight‐nucleotide region of the identified cis‐regulatory element is present in the promoters of these four AMP genes. These studies suggest that 20‐hydroxyecdysone regulates the expression of AMP genes through a conserved cis‐regulatory element.
Shotgun proteomics approach to characterizing the embryonic proteome of the silkworm, Bombyx mori, at labrum appearance stage
Authors: Li J‐Y. , Chen X. , Hosseini Moghaddam S. H. , Chen M. , Wei H. , Zhong B‐X.
Published: September 2009

The shotgun approach has gained considerable acknowledgement in recent years as a dominant strategy in proteomics. We observed a dramatic increase of specific protein spots in two‐dimensional electrophoresis (2‐DE) gels of the silkworm (Bombyx mori) embryo at labrum appearance, a characteristic stage during embryonic development of silkworm which is involved with temperature increase by silkworm raiser. We employed shotgun liquid chromatography tandem mass spectrometry (LC‐MS/MS) technology to analyse the proteome of B. mori embryos at this stage. A total of 2168 proteins were identified with an in‐house database. Approximately 47% of them had isoelectric point (pI) values distributed theoretically in the range pI 5–7 and approximately 60% of them had molecular weights of 15–45 kDa. Furthermore, 111 proteins had an pI greater than 10 and were difficult to separate by 2‐DE. Many important functional proteins related to embryonic development, stress response, DNA transcription/translation, cell growth, proliferation and differentiation, organogenesis and reproduction were identified. Among them proteins related to nervous system development were noticeable. All known heat shock proteins (HSPs) were detected in this developmental stage of B. mori embryo. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed energetic metabolism at this stage. These results were expected to provide more information for proteomic monitoring of the insect embryo and better understanding of the spatiotemporal expression of genes during embryonic developmental processes.
Changes in gene expression in the permissive larval host lightbrown apple moth (Epiphyas postvittana, Tortricidae) in response to EppoNPV (Baculoviridae) infection
Authors: Gatehouse H. S. , Poulton J. , Markwick N. P. , Gatehouse L. N. , Ward V. K. , Young V. L. , Luo Z. , Schaffer R. , Christeller J. T.
Published: September 2009

Host cell and virus gene expression were measured five days after per os inoculation of 3rd instar lightbrown apple moth (LBAM) larvae with the Epiphyas postvittana nucleopolyhedrovirus (EppoNPV). Microarray analysis identified 84 insect genes that were up‐regulated and 18 genes that were down‐regulated in virus‐infected larvae compared with uninfected larvae. From the 134 viral open reading frames represented on the microarray, 81 genes showed strong expression. Of the 38 functionally identifiable regulated insect genes, 23 coded for proteins that have roles in one of five processes; regulation of transcription and translation, induction of apoptosis, and maintenance of both juvenility and actin cytoskeletal integrity. Of the 34 functionally identifiable viral genes that were most strongly expressed, 12 had functions associated with these five processes, as did a further seven viral genes which were expressed at slightly lower levels. A survey of the LBAM‐expressed sequence tag library identified further genes involved in these processes. In total, 135 insect genes and 38 viral genes were analysed by quantitative polymerase chain reaction. Twenty‐one insect genes were strongly up‐regulated and 31 genes strongly down‐regulated. All 38 viral genes examined were highly expressed. These data suggest that induction of apoptosis and regulation of juvenility are the major ‘battlegrounds’ between virus and insect, with the majority of changes observed representing viral control of insect gene expression. Transcription and translational effects seem to be exerted largely through modulation of mRNA and protein degradation. Examples of attempts by the insect to repel the infection via changes in gene expression within these same processes were, however, also noted. The data also showed the extent to which viral transcription dominated in the infected insects at five days post inoculation.
Drosomycin, an essential component of antifungal defence in Drosophila
Authors: Zhang Z‐T. , Zhu S‐Y.
Published: September 2009

Drosomycin is an inducible antifungal peptide of 44 residues initially isolated from bacteria‐challenged Drosophila melanogaster. The systemic expression of drosomycin is regulated by the Toll pathway present in fat body, whereas inducible local expression in the respiratory tract is controlled by the Immune Deficiency (IMD) pathway. Drosomycin belongs to the cysteine‐stabilized α‐helical and β‐sheet (CSαβ) superfamily and is composed of an α‐helix and a three‐stranded β‐sheet stabilized by four disulphide bridges. Drosomycin exhibits a narrow antimicrobial spectrum and is only active against some filamentous fungi. However, recent work using recombinant drosomycin expressed in Escherichia coli revealed its antiparasitic and anti‐yeast activities. Two evolutionary epitopes (α‐ and γ‐patch) and the m‐loop have been proposed as putative functional regions of drosomycin for interaction with fungi and parasites, respectively. Similarity in sequence, structure and biological activity suggests that drosomycin and some defensin molecules from plants and fungi could originate from a common ancestor.