title: Epidemic and bioterrorism preparation among emergency medical services systems.

text: INTRODUCTION The purpose of this research was to determine the preparedness of emergency medical services (EMS) agencies in one US state to cope with a massive epidemic event. METHODS Data were collected primarily through telephone interviews with EMS officials throughout the State. To provide a comparison, nine out-of-state emergency services agencies were invited to participate. RESULTS Emergency medical services agencies from nine of the 23 counties (39%) provided responses to some or all of the questions in the telephone survey. Seven of the nine out-of-state agencies provided responses to the survey. Most of the EMS agencies do not have broad, formal plans for response to large-scale bio-terrorist or pandemic events. CONCLUSIONS The findings indicate that EMS agencies in this state fundamentally are unprepared for a large-scale bioterrorism or pandemic event. The few existing plans rely heavily on mutual aid from agencies that may be incapable of providing such aid. Therefore, EMS agencies must be prepared to manage a response to these incidents without assistance from any agencies outside of their local community. In order to accomplish this, they must begin planning and develop close working relationships with public health, healthcare, and elected officials within their local communities.


Rotavirus and coronavirus associated diarrhoea in domestic animals.


title: Host factors in the replication of positive-strand RNA viruses.

text: Viruses are obligate, intracellular parasites that depend on host cells for successful propagation. Upon infection of host cells, positivestrand RNA viruses exploit and hijack cellular machinery and reprogram these cells into viral "factories" through various protein-protein, protein- RNA, and protein-lipid interactions. The molecular interplay between host factors and invading viruses is a continuous process throughout the entire viral life cycle and determines virus host range and viral pathogenesis, as well as driving viral evolution. Studies of host factors have contributed insights into their normal cellular functions and helped identify attractive targets for antiviral drug development. With the development of high throughput screening, functional genomics, and proteomics technologies, host factors participating in viral life cycles have been identified rapidly in recent years. In this review, we summarize the recent advances in virus-host cell interactions in positive-strand RNA virus infections and focus on host factors that facilitate viral replication.


title: [Infections with the MERS coronavirus--for the present no threat to Europe].

text: In Saudi Arabia, a novel coronavirus named Middle East respiratory syndrome coronavirus (MERS-CoV) was isolated in 2012 from patients with severe respiratory symptoms. Up to now, more than 1600 MERS cases have been registered mainly in the Arabian Peninsula. MERS is usually accompanied with fever, cough, and shortness of breath. In many cases, pneumonia is observed. However, clinical features of MERS range from mild disease to acute respiratory distress syndrome and multiorgan failure resulting in death, especially in individuals with underlying comorbidities. To date, about one in three people died as a result of MERS. In Europe, MERS cases have only been registered in isolated travelers entering from the Middle East.


title: Genome of infectious bronchitis virus.

text: Techniques are described for the growth and rapid purification of the avian coronavirus infectious bronchitis virus (IBV). Purified IBV has a sedimentation coefficient of 320S and a buoyant density of 1.22 g/ml in sucrose-deuterium oxide equilibrium gradients. IBV RNA extracted by proteinase K in the presence of sodium dodecyl sulfate and further purified by phenol extraction and gradient centrifugation is single stranded and has a sedimentation coefficient of 64S, as determined by isokinetic gradient centrifugation. Analysis on sucrose gradients under both aqueous and denaturing conditions together with agarose gel electrophoresis in the presence of the chaotropic agent methylmercuric hydroxide gave a value of 8 X 10(6) for the moleclar weight of IBV RNA. This value was confirmed by RNase T1 fingerprinting, which also indicated that IBV RNA is haploid. No evidence was found of subunit structure in IBV RNA. From these results together with the recently reported observation that IBV RNA is infectious and contains a tract of polyadenylic acid (Lomniczi, J. Gen. Virol., in press), we conclude that the genome of the coronaviruses is a single continuous chain of about 23,000 mononucleotides that is of messenger polarity.


title: Fusogenic properties of uncleaved spike protein of murine coronavirus JHMV.

text: We have tested the fusogenic properties of cleaved and uncleaved spike (S) protein of murine coronavirus (MCV) JHMV variant cl-2 by expressing the S protein by recombinant vaccinia viruses (RVVs). The amino acid sequence of the putative cleavage site of cl-2 S protein, Arg-Arg-Ala-Arg-Arg, was replaced by Arg-Thr-Ala-Leu-Glu by in vitro mutagenesis of cl-2 S gene. The RVVs having cl-2 S gene [RVV t(+)] or mutated cl-2 S gene [RVV t(-)] were tested for their ability to induce fusion as well as cleavability in DBT cells. After inoculation with RVV t(+) onto DBT cells, the fusion formation was first observed at 8 h postinoculation (p.i.) and spread throughout the whole culture by 24 h. In cells infected with RVV t(-), fusion appeared by 2 h and most of cells were fused by 30 h p.i. The S protein and its cleavage products were detected in DBT cells expressing wild type S protein. However, no cleavage products of the S protein were detected in RVV t(-) infected cells producing mutated S protein, even though fusion was clearly visible. These results suggest that the cleavage event of JHMV-S protein of MCV is not a prerequisite for fusion formation, but that it enhances fusion.


Statement of ESMINT and ESNR regarding recent trials evaluating the endovascular treatment at the acute stage of ischemic stroke.


docid: xsvmijac

title: Recombinant herpesviruses as tools for the study of herpesvirus biology.

text: This article is a brief summary of efforts to generate mutant herpesviruses for investigating and assigning gene functions of herpesviruses in replication and pathogenesis. While a full review of all herpesviruses is beyond the scope of this review, we focused our attention on the prototype of the herpesvirus subfamily - herpes simplex virus and murine gammaherpesvirus that serves as an excellent animal model to study human gammaherpesvirus pathogenesis. Furthermore, our present knowledge of essential, non-essential, and common genes of herpesviruses as well as of accessory genes that are currently being studied with the help of the bacterial artificial chromosome (BAC) system will also be discussed. This system facilitates the analysis of herpesviral genes with potential for use in gene therapy or as anti-cancer therapeutics.


docid: kbwobrqu

title: Outbreak of E. coli O157:H7 associated with bathing at a public beach in the Montreal-Centre region.


title: Clinical analysis of 45 patients with severe acute respiratory syndrome.

text: OBJECTIVE To explore the clinical and radiological features of severe acute respiratory syndrome (SARS). METHODS Analysis of the clinical presentation, chest radiographs, course of disease and features of severe pneumonia in 45 SARS patients diagnosed at our hospital in Beijing between April 5 - 20, 2003. Also included is a summation of the clinical features of SARS. RESULTS (1) SARS appears to have high infectivity; (2) the most common symptom is fever; (3) the count of leukocyte is normal or decreased; (4) most patients (35/45, 77.8%) had experienced a 24-hour fever prior to the abnormal chest X-ray changes which showed progression of pulmonary infiltrates within 48 hours in 71.1% (32/45) of the patients and, (5) the percentage of patients who developed severe pneumonia (24.4%) is higher than those who developed typical pneumonia. CONCLUSION SARS is a disease with high infectivity and has its own clinical and radiological features. Early recognition, prompt isolation, and appropriate therapy are the key to combat this infection.


title: Exploratory rigid laparoscopy in an African elephant (Loxodonta africana).

text: In March 2009, a 25-yr-old captive female African elephant (Loxodonta africana) underwent an exploratory laparoscopy after several weeks of diarrhea, submandibular and ventral edema, and swelling on medial and lateral aspects of all feet. Although there have been recent advances in laparoscopic vasectomies in free-ranging African elephants in South Africa utilizing specially designed rigid laparoscopes and insufflation devices, this was the first attempt at using these same techniques for an exploratory purpose. The elephant was sedated in a static restraint chute and remained standing for the duration of the procedure. Laparoscopy provided visibility of the dorsal abdomen, enabled collection of reproductive tract biopsies and peritoneal fluid samples, and allowed for instillation of antibiotics and crystalloid fluids directly into the abdominal cavity. Abdominal exploration, collection of tissue samples, and local therapy is possible via standing laparoscopy in megavertebrates.


title: Flexibility as a Strategy in Nucleoside Antiviral Drug Design.

text: As far back as Melville Wolfrom's acyclic sugar synthesis in the 1960's, synthesis of flexible nucleoside analogues have been an area of interest. This concept, however, went against years of enzyme-substrate binding theory. Hence, acyclic methodology in antiviral drug design did not take off until the discovery and subsequent FDA approval of such analogues as Acyclovir and Tenofovir. More recently, the observation that flexible nucleosides could overcome drug resistance spawned a renewed interest in the field of nucleoside drug design. The next generation of flexible nucleosides shifted the focus from the sugar moiety to the nucleobase. With analogues such as Seley-Radtke "fleximers", and Herdewijn's C5 substituted 2'-deoxyuridines, the area of base flexibility has seen great expansion. More recently, the marriage of these methodologies with acyclic sugars has resulted in a series of acyclic flex-base nucleosides with a wide range of antiviral properties, including some of the first to exhibit anti-coronavirus activity. Various flexible nucleosides and their corresponding nucleobases will be compared in this review.


title: Notification of animal and human diseases: the global legal basis.

text: The successful control of a disease, and a possible epidemic, depends on rapid access to complete information on the disease situation. To ensure a timely response, diseases must be immediately notified in a transparent manner. The rapid exchange of information about animal diseases, including zoonoses, was the key objective in the establishment of the World Organisation for Animal Health (OIE) in 1924. For diseases concerning humans, a set of new rules dealing with the quick reporting of infectious diseases--the International Health Regulations--was adopted by Members of the World Health Organization (OMS) in 2005. The article explains these two systems of notification, which make information accessible to the public and allow decision-makers to better manage the risks related to the diseases concerned.


title: Intubation of SARS patients: infection and perspectives of healthcare workers.

text: PURPOSE The outbreak of severe acute respiratory syndrome (SARS) in 2003 presented major challenges to the safety of anesthesiologists and other healthcare workers (HCWs). This study determined the incidence of SARS transmission to HCWs who intubated patients and analyzed the concerns of HCWs regarding personal and patient safety. METHODS Healthcare workers who performed tracheal intubation in 10 Toronto hospitals were identified using the Ontario Public Health database. A questionnaire was used to collect information from the HCWs. To determine if the patterns of personal protection or concerns changed over time, data were analyzed according to whether the intubation occurred during SARS 1 (February 23 to April 21) or SARS 2 (April 22 to July 1). RESULTS Thirty-three HCWs who performed 39 intubations on 35 SARS patients were interviewed. Three of 23 HCWs (13%) acquired SARS during SARS 1 whereas none (0/10) acquired SARS during SARS 2. Personal protection increased from SARS 1 to SARS 2 and HCWs' concerns changed over time. During SARS 1, concerns focused on the need for personal protective equipment whereas during SARS 2, concerns focused on the need for strict training and patient care protocols. HCWs perceived that their experiences were ineffectively integrated into risk management protocols. CONCLUSIONS Protection guidelines failed to completely prevent the transmission of SARS to HCWs. Nine percent of the interviewed HCWs who intubated patients contracted SARS. A Risk Analysis Framework is presented to facilitate the rapid integration of HCWs' experiences into practice guidelines.


docid: gaepyggp

title: An investigation into the health status and diseases of wild dogs (Lycaon pictus) in the Kruger National Park.

text: Many factors have been cited as possible reasons for the decline in the number of wild dogs (Lycaon pictus), but few studies have provided supportive data. Between 1990 and 1993, the dynamics of 10 wild dog packs in the southern district of the Kruger National Park in South Africa were monitored. Casual observations of the causes of disease and mortality in the entire population were also recorded. During the same period, 46 wild dogs were immobilised, weighed, and subjected to physical examination as well as the collection of blood and faecal specimens. External parasites were collected and blood smears were prepared. Serum specimens were subjected to routine blood chemistry analyses, hormone and serological assays and formalinised blood specimens and faeces were subjected to screening for endoparasites. The study population varied from 75 in 1990 to 123 in 1993 with a survival rate of 29.9% for pups, 64.3% for yearlings and 69% for adults. Eighty per cent or more of the population were under 4 years of age. The cause of death in all wild dogs in the Kruger National Park could be established only in a small number of cases. Lions were responsible for the death of 20/62 wild dogs and disease caused the death of 6/62 wild dogs. All immobilised dogs were in a good physical condition, but 85% of dogs had one or more skin lesions. Potential life-threatening lesions (bitewounds inflicted by other dogs and lesions inflicted by a snare) occurred in 4 dogs. One male dog had only one testicle in the scortum. Twenty-six (93%) blood smears were positive for gametocytes of Hepatozoon sp., presumably H. canis, and in 2 dogs trophozoites of Babesia canis were seen. Eighty-six per cent of the specimens were positive for Dipetalonema reconditum. All dogs were infested with ticks and Haemaphysalis leachi, Amblyomma hebraeum, A. marmoreum, Boophilus decoloratus, Rhipicephalus simus, R. evertsi, R. appendiculatus and R. zambesiensis were identified. Ctenocephalides sp. and Echidnophaga larina were also identified. Taenia sp., Toxascaris canis and Ancylostoma caninum were present in faecal specimens. Antibody titres to adenovirus (26/31), B. canis (6/15), canine para-influenza virus (21/31), coronavirus (20/31), Coxiella burnetti (8/29), reovirus Type 3 (9/31), Rickettsia conori/africae (27/29), rotavirus (16/31) and Toxoplasma gondii (16/16) were found. The average serum urea concentration was higher (16.4 mmol/e) than that described for captive wild dogs, but other biochemical parameters were generally in agreement with values reported for captive wild dogs.(ABSTRACT TRUNCATED AT 400 WORDS)


title: COVID-19, cytokines and immunosuppression: what can we learn from severe acute respiratory syndrome?

text: A severe outbreak of coronavirus disease 2019 (COVID-19) emerged in China in December 2019, and spread so rapidly that more than 200,000 cases have so far been reported worldwide; on January 30, 2020, the WHO declared it the sixth public health emergency of international concern. The two previously reported coronavirus epidemics (severe acute respiratory syndrome [SARS] and Middle East respiratory syndrome [MERS]) share similar pathogenetic, epidemiological and clinical features as COVID-19. As little is currently known about SARS-CoV-2, it is likely that lessons learned from these major epidemics can be applied to the new pandemic, including the use of novel immunosuppressive drugs.


The 2014 Hajj and Umrah - current recommendations.


title: Identification of human aminopeptidase O, a novel metalloprotease with structural similarity to aminopeptidase B and leukotriene A4 hydrolase.

text: We have cloned and characterized a human brain cDNA encoding a new metalloprotease that has been called aminopeptidase O (AP-O). AP-O exhibits a series of structural features characteristic of aminopeptidases, including a conserved catalytic domain with a zinc-binding site (HEXXHX18E) that allows its classification in the M1 family of metallopeptidases or gluzincins. The structural complexity of AP-O is further increased by the presence of an additional C-terminal domain 170 residues long, which is predicted to have an ARM repeat fold originally identified in the Drosophila segment polarity gene product Armadillo. This ARM repeat domain is also present in aminopeptidase B, aminopeptidase B-like, and leukotriene A4 hydrolase and defines a novel subfamily of aminopeptidases that we have called ARM aminopeptidases. Northern blot analysis revealed that AP-O is mainly expressed in the pancreas, placenta, liver, testis, and heart. Human AP-O was produced in Escherichia coli, and the purified recombinant protein hydrolyzed synthetic substrates used for assaying aminopeptidase activity. This activity was abolished by general inhibitors of metalloproteases and specific inhibitors of aminopeptidases. Recombinant AP-O also cleaved angiotensin III to generate angiotensin IV, a bioactive peptide of the renin-angiotensin pathway with multiple actions on diverse tissues, including brain, testis, and heart. On the basis of these results we suggest that AP-O could play a role in the proteolytic processing of bioactive peptides in those tissues where it is expressed.


title: New York City's healthcare transportation during a disaster: a preparedness framework for a wicked problem.

text: During a disaster, victims with varied morbidities are located at incident sites, while healthcare facilities with varied healthcare resources are distributed elsewhere. Transportation serves an essential equilibrating role: it helps balance the patients' need for care with the supply of care. Studying the special case of New York City, this article sets out the healthcare transportation components as: (1) incident morbidity; (2) transportation assets; and (3) healthcare capacity. The relationship between these three components raises an assignment problem: the management of healthcare transportation within a dynamic and partly unpredictable incident-transportation-healthcare nexus, under urban disruption. While the routine dispatch problem can be tackled through better geographic allocation software and technical algorithms, the disaster assignment problem must be confronted through real-time, mutual adjustment between institutions. This article outlines institutional alternatives for managing the assignment problem and calls for further research on the merits of alternative institutional models.


title: Measures to Predict The Individual Variability of Corticospinal Responses Following Transcranial Direct Current Stimulation.

text: Individual responses to transcranial direct current stimulation (tDCS) are varied and therefore potentially limit its application. There is evidence that this variability is related to the contributions of Indirect waves (I-waves) recruited in the cortex. The latency of motor-evoked potentials (MEPs) can be measured through transcranial magnetic stimulation (TMS), allowing an individual's responsiveness to tDCS to be determined. However, this single-pulse method requires several different orientations of the TMS coil, potentially affecting its reliability. Instead, we propose a paired-pulse TMS paradigm targeting I-waves as an alternative method. This method uses one orientation that reduces inter- and intra-trial variability. It was hypothesized that the paired-pulse method would correlate more highly to tDCS responses than the single-pulse method. In a randomized, double blinded, cross-over design, 30 healthy participants completed two sessions, receiving 20 min of either anodal (2 mA) or sham tDCS. TMS was used to quantify Short interval intracortical facilitation (SICF) at Inter stimulus intervals (ISIs) of 1.5, 3.5 and 4.5 ms. Latency was determined in the posterior-anterior (PA), anterior-posterior (AP) and latero-medial (LM) coil orientations. The relationship between latency, SICF measures and the change in suprathreshold MEP amplitude size following tDCS were determined with Pearson's correlations. TMS measures, SICI and SICF were also used to determine responses to Anodal-tDCS (a-tDCS). Neither of the latency differences nor the SICF measures correlated to the change in MEP amplitude from pre-post tDCS (all P > 0.05). Overall, there was no significant response to tDCS in this cohort. This study highlights the need for testing the effects of various tDCS protocols on the different I-waves. Further research into SICF and whether it is a viable measure of I-wave facilitation is warranted.


title: A cross-sectional study on reference ranges of normal oral temperatures among students in Singapore.

text: INTRODUCTION During the 2003 Severe Acute Respiratory Syndrome (SARS) outbreak, all schools in Singapore implemented twice-daily temperature monitoring for students to curtail the spread of the disease. Students were not allowed to attend school if their temperature readings were >37.8 degrees C for students < or =12 years old, or > or =37.5 degrees C for students >12 years old. These values had been arbitrarily determined with professional inputs. The aim of this study is to determine the reference ranges of normal oral temperatures of students in Singapore and recommend the cut-off values for febrile patients. This may be used in another similar outbreak of an infectious disease with fever. MATERIALS AND METHODS Four co-ed primary schools and 4 co-ed secondary schools were selected for this study. Four thousand and two hundred primary 1 to secondary 3 students responded (96.8%) and participated in this cross-sectional study. The mean ages of the students in the lowest (primary 1) and highest educational levels (secondary 3) were 7.4 years old and 15.3 years old, respectively. Twelve oral temperature readings per student (i.e. measurements taken 4 times a day in 3 consecutive days) were collected. Forty-six thousand seven hundred and eighty-three (92.8%) out of 50,400 temperature readings were used for the analysis as missing data were excluded. A quantile regression model was applied to estimate reference ranges of normal oral temperatures for students with adjustment for potential confounding factors. RESULTS The age-specific reference ranges of normal oral temperature from this study for students < or =12 years old and >12 years old were 35.7 degrees C to 37.7 degrees C and 35.6 degrees C to 37.4 degrees C, respectively. Temperatures of 37.8 degrees C and 37.5 degrees C are therefore recommended as the oral temperature cut-offs for those < or =12 years old and >12 years old, respectively. CONCLUSION This study has provided empirical data on normal oral temperature cut-offs which could be used during temperature screening in schools.


title: Severe respiratory illness caused by a novel coronavirus, in a patient transferred to the United Kingdom from the Middle East, September 2012.

text: Coronaviruses have the potential to cause severe transmissible human disease, as demonstrated by the severe acute respiratory syndrome (SARS) outbreak of 2003. We describe here the clinical and virological features of a novel coronavirus infection causing severe respiratory illness in a patient transferred to London, United Kingdom, from the Gulf region of the Middle East.


title: Programmed translational frameshifting is likely required for expressions of genes encoding putative nuclear protein kinases of the ciliate Euplotes octocarinatus.

text: Three macronuclear genes encoding putative nuclear protein kinases of the ciliate Euplotes octocarinatus syngen 1 were isolated and sequenced. All three deduced gene products share significant properties with a group of recently identified nuclear serine/threonine protein kinases named Ndr. The three predicted proteins contain the twelve conserved catalytic subdomains of protein kinases and 22 near universally-conserved amino acids residues that are characteristic of serine/threonine protein kinases. In addition, there is an approximately 30 amino acid-peptide insertion between subdomains VII and VIII that contains a potential nuclear localization signal. Sequence analysis suggests that expression of the Eondr2 gene requires a + 1 programmed translational frameshift for its translation. Comparison of the deduced EoNdr2 with other known Ndr protein kinases implies that a + 1 ribosomal frameshift occurs at the motif AAATAA.


Does screening keep Ebola out of USA?


Current understanding of vaccination site-associated sarcomas in the cat.


title: Ecological origins of novel human pathogens.

text: A systematic literature survey suggests that there are 1399 species of human pathogen. Of these, 87 were first reported in humans in the years since 1980. The new species are disproportionately viruses, have a global distribution, and are mostly associated with animal reservoirs. Their emergence is often driven by ecological changes, especially with how human populations interact with animal reservoirs. Here, we review the process of pathogen emergence over both ecological and evolutionary time scales by reference to the "pathogen pyramid." We also consider the public health implications of the continuing emergence of new pathogens, focusing on the importance of international surveillance.


title: Multiple receptor-dependent steps determine the species specificity of HCV-229E infection.

text: Human coronavirus (HCV)-229E causes disease only in humans and grows in human cells and in cells of other species that express recombinant human aminopeptidase N (hAPN), the receptor for HCV-229E. We compared the species specificity of HCV-229E infection with the species specificity of virus binding using immunofluorescence, assay of virus yields, fluorescence activated cell sorting and a monoclonal antibody directed against hAPN that blocks infection. We found that HCV-229E binds to intestinal brush border membranes (BBM) and to membranes of cell lines from cats, dogs, pigs, and humans, however the virus only infects two of these species. HCV-229E will not bind to BBM or to membranes from cell lines derived from hamster or mice. Animal coronaviruses related to HCV-229E, including FIPV, CCV, and TGEV bind to cell membranes from cats, dogs, cows, pigs and humans (but not mice), while each virus infects cells from only a subset of these species. Infectious genomic HCV-229E RNA, can infect cells of all of these species. These data suggest that the species-specificity of infection for this serogroup of coronaviruses is determined at the levels of virus binding and penetration. Since binding of viral spike glycoprotein to cellular receptors is not the only limiting factor, we suggest that one or more steps associated with virus penetration may determine the species specificity of infection with the HCV-229E serogroup of coronaviruses.


title: Astrovirus induces diarrhea in the absence of inflammation and cell death.

text: Astroviruses are a leading cause of infantile viral gastroenteritis worldwide. Very little is known about the mechanisms of astrovirus-induced diarrhea. One reason for this is the lack of a small-animal model. Recently, we isolated a novel strain of astrovirus (TAstV-2) from turkeys with the emerging infectious disease poult enteritis mortality syndrome. In the present studies, we demonstrate that TAstV-2 causes growth depression, decreased thymus size, and enteric infection in infected turkeys. Infectious TAstV-2 can be recovered from multiple tissues, including the blood, suggesting that there is a viremic stage during infection. In spite of the severe diarrhea, histopathologic changes in the intestine were mild and there was a surprising lack of inflammation. This may be due to the increased activation of the potent immunosuppressive cytokine transforming growth factor beta during astrovirus infection. These studies suggest that the turkey will be a useful small-animal model with which to study astrovirus pathogenesis and immunity.


docid: zlcndqhm

title: Organizational aspects of intensive care unit resource allocation in a primary respiratory intensive care center specialized for the treatment of SARS-COV-2 patients.


title: Broad cellular immunity with robust memory responses to simian immunodeficiency virus following serial vaccination with adenovirus 5- and 35-based vectors.

text: Adenovirus serotype 35 (Ad35) is a promising vaccine platform for human immunodeficiency virus (HIV) infection and emerging infectious diseases as it is uncommon in humans worldwide and is distinct from Ad5, the major vaccine serotype for which many individuals have pre-existing immunity. The immunogenicity of a first-generation, replication-competent Ad35-based vaccine was tested in the simian immunodeficiency virus (SIV) rhesus macaque model by evaluating its capacity to boost immunity generated by Ad5-based vectors. A series of four immunizations with replication-defective Ad5 vectors expressing SIVmac239 gag induced high-frequency responses mediated by both CD8(+) and CD4(+) T cells directed against several epitopes. Ad5-specific neutralizing antibody responses that did not neutralize Ad35 were rapidly induced but waned over time. Subsequent immunization with Ad5-based vectors was minimally effective, whereas immunization with Ad35-based vectors generated a strong increase in the frequency of Gag-specific T cells with specificities that were unchanged. While this boosting response was relatively transient, challenge with the distinct pathogenic isolate SIV/DeltaB670 generated robust and selective recall responses to Gag with similar specificities as induced by vaccination that were elevated for 25 weeks relative to controls. Vaccination had measurable albeit minor effects on virus load. Unexpectedly, regional hypervariability within the Gag sequence of SIV/DeltaB670 was associated with mutation of the conserved CD8(+) T-cell epitope CM9 without concurrent flanking mutations and in the absence of immune pressure. These findings support the further development of Ad35 as a vaccine vector, and promote vaccine regimens that utilize serial administration of heterologous adenoviruses.


docid: jgdqtxnz

title: [The expression and activity detection of a variant N protein of SARS-CoV].

text: AIM To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV and GST in E.coli. METHODS The N region gene of SARS-CoV was cloned by RT-PCR. The expression vector was constructed by DNA recombination. The recombinant plasmid was transformed into E.coli BL21(DE3). The expression of the fusion protein was detected by Western blot. RESULTS (1) As compared with the sequences in GenBank, 20 bp were deleted in DNA sequence of the cloned N protein. (2) The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive. CONCLUSION The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, which lays the foundation for further study of SARS-CoV N protein.


title: An office-based approach to influenza: clinical diagnosis and laboratory testing.

text: Vaccination is the primary measure for preventing morbidity and mortality from influenza. During the influenza season, family physicians must distinguish influenza from the common cold and other flu-like illnesses. Signs and symptoms of influenza include abrupt onset of fever, severe myalgias, anorexia, sore throat, headache, cough, and malaise. Clinical diagnosis can be difficult or nonspecific when patients have other symptoms (e.g., stuffy nose, sneezing, cough, sore throat) that can be caused by various respiratory viruses or bacterial pathogens. Family physicians can improve diagnostic accuracy by being aware of the epidemiology of influenza. During outbreaks of influenza, commercially available rapid assays can be used to identify type A and B viruses. On average, rapid in-office tests are more than 70 percent sensitive and 90 percent specific for viral antigens. The assays vary in complexity, specificity, sensitivity, time to obtain results, specimen analyzed, and cost. The results of rapid viral tests can guide treatment decisions.


title: Nonassociative explanations of behavioral changes induced by central cholinergic drugs.

text: Studies concerning the effects of cholinergic stimulants and blockers on habituation, alternation, classical and instrumental conditioning, differentiation, and discrimination are summarized and discussed. This analysis suggests that many results originally ascribed to associative changes can be more economically accounted for by nonassociative factors inherent in the experiments. Furthermore, the data show that neither one of the more economical nonassuciative explanations at either the sensory, the drive, or the response level can by itself account for the complex interactions observed. The studies dealing with drug effects in lesioned animals, and those using intracerebral treatments, point out several sites of action of muscarinic agonists and antagonists in the septum, the hippocampus, the amygdala, the striatum, the hypothalamus, midline thalamic nuclei, and the reticular formation. Therefore, the provisional conclusion must be that central muscarinic systems subserve several functions, ranging from the modulation of consummatory responses to the control of motor-act differentiation. Both the drug and the lesion studies show not only that the fronto-limbic system has a considerable role in the latter process, but also that the localization of motor-act differentiation under a given drive depends both on the category of the cue, and on the category of the response. Some comparisons between the effects of muscarinic agonists and blockers and those of other drugs indicate that the alteration of sensory-motor relations caused by the former can be separated from (i) the motor hyperactivity caused by amphetamine, which is relatively less dependent on sensory factors; (ii) the selective (response enhancements caused by the indole hallucinogen LSD25, which appear to be amenable to a disinhibition of drive; (iii) the attenuation of punishment suppression by sedative and antianxiety agents (benzodiazepines and subhypnotic doses of barbiturates); and (iv) the hyperarousal or hyperreactivity syndrome induced by compounds which deplete central 5-hydroixytryphamine stores, such as panachlorophenylalanine.


title: The possible use of combined electrical impedance and ultrasound velocity measurements for the non-invasive measurement of temperature during mild hyperthermia.

text: This paper explores the possibility of using combined measurements of electrical impedance and changes in ultrasound time of flight for determining deep body temperature during mild hyperthermia. Simultaneous electrical impedance spectra (1 kHz-1024 kHz) and ultrasound time-of-flight measurements were made on layered sheep liver and fat tissue samples as the temperature was increased from 30-50 °C. The change in propagation velocity for 100% fat and 100% liver samples was found to vary linearly with temperature and the temperature coefficient of the time-of-flight was shown to vary linearly with the % fat in the sample (0.009% °C-1%-1). Tetrapolar impedance measurements normalized to 8 kHz were shown to have a small sensitivity to temperature for both liver (0.001% °C-1 ≤ 45 °C) and fat (0.002% °C-1 ≤ 512 kHz) and the best linear correlation between the normalized impedance and the % fat in the sample was found at 256 kHz (gradient 0.026%-1, r2 = 0.65). A bootstrap analysis on 15 layered tissue samples evaluated using the normalized impedance at 256 kHz to determine the % fat in the sample and the temperature coefficient of the time of flight to determine the temperature. The results showed differences (including some large differences) between the predicted and measured temperatures and an error evaluation identified the possible origins of these.


title: Rapid and sensitive detection of severe acute respiratory syndrome coronavirus by rolling circle amplification.

text: The severe acute respiratory syndrome (SARS) epidemic of 2003 was responsible for 774 deaths and caused significant economic damage worldwide. Since July 2003, a number of SARS cases have occurred in China, raising the possibility of future epidemics. We describe here a rapid, sensitive, and highly efficient assay for the detection of SARS coronavirus (SARS-CoV) in cultured material and a small number (n = 7) of clinical samples. Using rolling circle amplification (RCA), we were able to achieve sensitive detection levels of SARS-CoV RNA in both solid and liquid phases. The main advantage of RCA is that it can be performed under isothermal conditions with minimal reagents and avoids the generation of false-positive results, a problem that is frequently encountered in PCR-based assays. Furthermore, the RCA technology provides a faster, more sensitive, and economical option to currently available PCR-based methods.


title: Effect of 2 Bedding Materials on Ammonia Levels in Individually Ventilated Cages.

text: This study sought to identify an optimal rodent bedding and cage-change interval to establish standard procedures for the IVC in our rodent vivarium. Disposable cages were prefilled with either corncob or α-cellulose bedding and were used to house 2 adult Sprague-Dawley rats (experimental condition) or contained no animals (control). Rats were observed and intracage ammonia levels measured daily for 21 d. Intracage ammonia accumulation became significant by day 8 in experimental cages containing α-cellulose bedding, whereas experimental cages containing corncob bedding did not reach detectable levels of ammonia until day 14. In all 3 experimental cages containing α-cellulose, ammonia exceeded 100 ppm (our maximum acceptable limit) by day 11. Two experimental corncob cages required changing at days 16 and 17, whereas the remaining cage containing corncob bedding lasted the entire 21 d without reaching the 100-ppm ammonia threshold. These data suggests that corncob bedding provides nearly twice the service life of α-cellulose bedding in the IVC system.


Prevalence of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus infection in Korean pigs.


title: Infection with cytotoxic T-lymphocyte escape mutants results in increased mortality and growth retardation in mice infected with a neurotropic coronavirus.

text: C57BL/6 mice infected with mouse hepatitis virus strain JHM (MHV-JHM) develop a chronic demyelinating encephalomyelitis several weeks after inoculation. Previously, we showed that mutations in the immunodominant CD8 T-cell epitope (S-510-518) could be detected in nearly all samples of RNA and virus isolated from these mice. These mutations abrogated recognition by T cells harvested from the central nervous systems of infected mice in direct ex vivo cytotoxicity assays. These results suggested that cytotoxic T-lymphocyte (CTL) escape mutants contributed to virus amplification and the development of clinical disease in mice infected with wild-type virus. In the present study, the importance of these mutations was further evaluated by infecting naive mice with MHV-JHM variants isolated from infected mice and in which epitope S-510-518 was mutated. Compared to mice infected with wild-type virus, variant virus-infected animals showed higher mortality and morbidity manifested by decreased weight gain and neurological signs. Although a delay in the kinetics of virus clearance has been demonstrated in previous studies of CTL escape mutants, this is the first illustration of significant changes in clinical disease resulting from infection with viruses able to evade the CD8 T-cell immune response.


Proposed new International Health Regulations.


docid: antmwagm

title: Monoclonal antibody analysis of neutralization and antibody-dependent enhancement of feline infectious peritonitis virus.

text: Fifty-four monoclonal antibodies (MAbs) to feline infectious peritonitis virus (FIPV) were characterized according to protein specificity, immunoglobulin subclass, virus neutralization, reactivity with different coronaviruses, and ability to induce antibody-dependent enhancement (ADE) of FIPV infection in vitro. The MAbs were found to be specific for one of three structural proteins of FIPV. A total of 47 MAbs were specific for the 205-kDa spike protein (S), 3 MAbs were specific for the 45-kDa nucleocapsid protein (N), and 4 MAbs were specific for the 26- to 28-kDa membrane protein (M). The S-specific MAbs showed various degrees of cross-reactivity with strains of FIPV, feline enteric coronavirus, canine coronavirus, and porcine transmissible gastroenteritis virus. Nineteen S-specific MAbs neutralized FIPV. A total of 15 of the neutralizing MAbs induced ADE, and all but 1 were of the immunoglobulin G2a subclass. The remaining four neutralizing MAbs that did not induce ADE were of the immunoglobulin G1 subclass. Two S-specific MAbs induced ADE but were nonneutralizing. None of the N- or M-specific MAbs was neutralizing or induced ADE. On the basis of the reactivity patterns of the MAbs with FIPV and related coronaviruses, it was concluded that there is a minimum of five neutralizing sites on S. In most instances, neutralizing MAbs were able to induce ADE, demonstrating a direct relationship between neutralization and enhancement. The difference in immunoglobulin subclass between neutralizing MAbs that induced ADE and those that did not induce ADE suggests that there may be a restriction in the immunoglobulin subclasses capable of mediating ADE.


title: Identification and primary structure of the gene encoding the Berne virus nucleocapsid protein.

text: The nucleotide sequence of the nucleocapsid (N) protein gene of Berne virus (BEV; proposed family Toroviridae) was determined from two independent clones of a cDNA library. From the deduced amino acid sequence a basic protein of 18.3K was predicted. In vitro transcription and translation, followed by immunoprecipitation, were used to identify the gene. The identification was confirmed by metabolic labelling, using the knowledge that cysteine residues are absent from the amino acid sequence of the N protein. Smaller N-related polypeptides encountered in BEV-infected cell lysates were shown to be probable products of aberrant translation, due to initiation on AUG codons further downstream in the N protein gene.


title: Protection of mice from lethal coronavirus MHV-A59 infection by monoclonal affinity-purified spike glycoprotein.

text: Numerous studies have provided indirect evidence that the spike glycoprotein of coronaviruses (E2 or S) bears determinants for pathogenesis and the induction of protective immunity. In order to directly evaluate its immunogenicity, the E2 glycoprotein of the murine hepatitis virus, strain A59, was purified by immunoaffinity chromatography. High titers of neutralizing and fusion inhibiting antibodies were induced in mice vaccinated with purified E2/S in Freund's adjuvant, which were protected from an intracerebral challenge with 10 LD50 of MHV-A59. This study provides a direct demonstration of the importance of the coronavirus spike glycoprotein in the induction of a protective immune response.


title: [Male violence against heterosexual partners: facts and challenges. Review of Mexican literature].

text: Violence against women by partners (VAWP) has been systematically studied in Mexico in the past 15 years. The aim of this paper is to reflect on the contributions on this topic in Mexico. The discussion focuses on the magnitude of this phenomenon, stressing its theoretical, methodological, and technical implications; the relationship between violence and a variety of factors; and the outcomes of VAWP on women themselves. The manuscript features the main contributions from quantitative and qualitative studies.


Inflammation and coagulation in the cardiovascular system: the contribution of influenza.


title: Comparison of the di- and trinucleotide frequencies from the genomes of nine different coronaviruses.

text: As an alternative to protein alignments for the comparison of sequences, the reiterations of mono- di- and trinucleotide frequencies were used for the comparison of coronavirus sequences. The relative abundance of the di- and trinucleotide frequencies within the 3' part from nine coronavirus genomes were determined. The patterns of dinucleotide frequencies and the trinucleotide frequencies showed some common features for all coronaviruses but also differences between the groups formerly defined on the base of antigenic relatedness. The normalised dinucleotide frequencies were further used to calculate the distances between coronavirus sequences. Based on the dinucleotide frequency distances, coronaviruses can be divided into two groups which roughly reflect the taxonomic groups. In this kind of evaluation, however, IBV occupies a position different to the one that it would take based on most protein sequence comparisons. Based on similarities within coding sequences and antigenic properties IBV occupies a place outside of both groups. Based on the dinucleotide frequencies IBV gained a position in between of the TGEV-related and the MHV-clustered coronaviruses.


docid: bvczlpqs

title: Influenza and other actue respiratory diseases in the Czech Socialist Republic, 1969-1974.

text: Since 1969, the incidence of acute respiratory diseases (ARD) in the Czech Socialist Republic of Czechoslovakia has been monitored by a special programme based on reports from 85 district epidemiological centres. In this paper, the incidence of ARD in three age groups, together with the incidence of complications and death rates, are presented for each season during the period 1969-1974. The significance of epidemiological observations and laboratory investigations relating to influenza and other ARD agents, such as parainfluenza viruses, adenoviruses, rhinoviruses, RS virus, coronaviruses, and Mycoplasma pneumoniae, is discussed.


title: Spatial clustering of SARS in Hong Kong.

text: 1. Geographic information system (GIS) can be applied during an acute infectious disease outbreak to reveal new geospatial information in addition to standard field epidemiological analyses. 2. When applied in real time during the onset and evolution of an epidemic, GIS can monitor and enhance understanding of the transmission dynamics of an infectious agent, thereby facilitating the design, implementation and evaluation of potential intervention strategies.


title: Feline infectious peritonitis in a cat presented because of papular skin lesions.

text: A 19-week-old neutered male domestic shorthair cat was examined because of multiple raised pruritic skin lesions along the dorsal head and back. Histopathology of biopsies of the lesions detected nodular pyogranulomatous dermatitis with vasculitis and necrosis, leading to a suspicion of feline infectious peritonitis (FIP). Postmortem examination revealed gross lesions consistent with FIP. Histopathologic lesions and positive immunohistochemical staining for feline coronavirus in multiple tissues, including the skin, confirmed the diagnosis of FIP. The current case was similar to previous cases, except for the initial presentation with cutaneous lesions and no other clinical signs, which had not been reported previously.


title: Virally activated CD8 T cells home to Mycobacterium bovis BCG-induced granulomas but enhance antimycobacterial protection only in immunodeficient mice.

text: The effect of secondary infections on CD4 T-cell-regulated chronic granulomatous inflammation is not well understood. Here, we have investigated the effect of an acute viral infection on the cellular composition and bacterial protection in Mycobacterium bovis strain bacille Calmette-Guérin (BCG)-induced granulomas using an immunocompetent and a partially immunodeficient murine model. Acute lymphocytic choriomeningitis virus (LCMV) coinfection of C57BL/6 mice led to substantial accumulation of gamma interferon (IFN-gamma)-producing LCMV-specific T cells in liver granulomas and increased local IFN-gamma. Despite traffic of activated T cells that resulted in a CD8 T-cell-dominated granuloma, the BCG liver organ load was unaltered from control levels. In OT-1 T-cell-receptor (TCR) transgenic mice, ovalbumin (OVA) immunization or LCMV coinfection of BCG-infected mice induced CD8 T-cell-dominated granulomas containing large numbers of non-BCG-specific activated T cells. The higher baseline BCG organ load in this CD8 TCR transgenic animal allowed us to demonstrate that OVA immunization and LCMV coinfection increased anti-BCG protection. The bacterial load remained substantially higher than in mice with a more complete TCR repertoire. Overall, the present study suggests that peripherally activated CD8 T cells can be recruited to chronic inflammatory sites, but their contribution to protective immunity is limited to conditions of underlying immunodeficiency.


title: Utility of the over-the-scope-clip system for treating a large esophageal perforation.

text: We report here a case of esophageal perforation made by an endoscope while treating cicatrical stenosis that developed after wide circumferential dissection of superficial esophageal carcinoma. Perforation closure with a conventional endoclip was difficult as the perforation was large and the surrounding tissue was fragile as a result of steroids administration for stenosis prevention. To avoid surgical intervention, we employed the over-the-scope-clip system and successfully closed the perforation. The favorable outcome suggests the utility of the over-the-scope-clip system for closing perforations when conventional methods are ineffective.


title: Structure-function relationships of the viral RNA-dependent RNA polymerase: fidelity, replication speed, and initiation mechanism determined by a residue in the ribose-binding pocket.

text: Studies of the RNA-dependent RNA polymerase (RdRp) from poliovirus (PV), 3Dpol, have shown that Asn-297 permits this enzyme to distinguish ribose from 2'-deoxyribose. All animal RNA viruses have Asn at the structurally homologous position of their polymerases, suggesting a conserved function for this residue. However, all prokaryotic RNA viruses have Glu at this position. In the presence of Mg2+, the apparent affinity of Glu-297 3Dpol for 2'-deoxyribonucleotides was decreased by 6-fold relative to wild type without a substantial difference in the fidelity of 2'-dNMP incorporation. The fidelity of ribonucleotide misincorporation for Glu-297 3Dpol was reduced by 14-fold relative to wild type. A 4- to 11-fold reduction in the rate of ribonucleotide incorporation was observed. Glu-297 PV was unable to grow in HeLa cells due to a replication defect equivalent to that observed for a mutant PV encoding an inactive polymerase. Evaluation of the protein-(VPg)-primed initiation reaction showed that only half of the Glu-297 3Dpol initiation complexes were capable of producing VPg-pUpU product and that the overall yield of uridylylated VPg products was reduced by 20-fold relative to wild-type enzyme, a circumstance attributable to a reduced affinity for UTP. These studies identify the first RdRp derivative with a mutator phenotype and provide a mechanistic basis for the elevated mutation frequency of RNA phage relative to animal RNA viruses observed in culture. Although protein-primed initiation and RNA-primed elongation complexes employ the same polymerase active site, the functional differences reported here imply significant structural differences between these complexes.


title: [Prevalence and seasonal distribution of respiratory viruses in patients with acute respiratory tract infections, 2002-2014].

text: The aim of this study was to investigate the prevalence and seasonal distribution of respiratory viruses in pediatric and adult outpatients and inpatients who were admitted to hospital with the symptoms of upper and lower respiratory tract infections, during a 12-year period. A total of 5102 clinical samples (4372 nasopharyngeal swabs, 316 bronchoalveolar lavages, 219 transtracheal aspirates, 163 nasopharyngeal aspirates, 20 sputum, 10 nasal swabs) examined in our laboratory between January 1st 2002 and July 17th 2014, were evaluated retrospectively. Of the specimens, 1107 (21.7%) were obtained from outpatients and 3995 (78.3%) from hospitalized patients. Of the patients, 2851 (55.9%) were male and 2251 (44.1%) were female, while 1233 (24.2%) were adults and 3869 (75.8%) were children (age range: 1 day - 93 years; median: 3 years). Respiratory samples were investigated for the presence of respiratory syncytial virus (RSV), influenza virus type A and B (INF-A, INF-B), adenovirus (AdV), parainfluenza viruses (PIV types 1-4), human rhinoviruses (HRV), human coronaviruses (HCoV), human metapneumovirus (HMPV) and human bocavirus (HBoV). All specimens were tested by both direct immunofluorescence antibody (DFA) and shell vial cell culture (SVCC) methods. In DFA assay the samples were initially screened by fluorescent-labeled polyclonal antibodies, and the positive ones were typed by using monoclonal antibodies (Light Diagnostics, Merck Millipore, USA). In SVCC, HEp-2, MDCK, A-549 and Vero cell lines were used for the isolation of viruses. In addition to these methods, real-time multiplex PCR methods (RealAccurate®, Respiratory RT PCR, PathoFinder, Netherlands and Seeplex® RV15 ACE Detection, Seegene, South Korea) were used for the detection of respiratory viruses in samples (n= 2104) obtained from 2007 to 2014. Respiratory viruses were detected in a total of 1705 (33.4%) patients, of them 967 (19%) were male and 738 (14.4%) were female. Three hundred and eighteen (18.6%) of the 1705 patients were infected with multiple respiratory viruses. The most frequently observed co-infections were RSV+INF-A (40/318; 12.6%), and RSV+PIV (33/318; 10.4%). The rate of positivity for the respiratory viruses in pediatric and adult groups were 35.4% (1369/3869) and 27.3% (336/1233), respectively (p< 0.000). The most frequently detected virus in pediatric group was RSV (336/1369; 24.5%), followed by influenza viruses (314/1369; 22.9%), PIV (197/1369; 14.4%), HRV (118/1369; 8.6%), AdV (75/1369; 5.5%) and the others (49/1369; 3.6%). On the other hand the most frequently detected virus in adult group was influenza viruses (181/336; 53.8%) followed by AdV (37/336; 11%), RSV (24/336; 7.1%), PIV (24/336; 7.1%), HRV (23/336; 6.8%) and the others (9/336; 2.7%). The rate of multiple virus infections in pediatric and adult groups were 7.2% (280/3869) and 3% (38/1233), respectively. Most of the coinfections (280/318; 88%) were detected in children. Respiratory viruses were detected positive in 40.2% (445/1107) of outpatients, and in 31.5% (1260/3995) of inpatients (p< 0.000). The most frequent viruses detected in pediatric outpatients and inpatients were HRV and RSV, respectively, while influenza viruses were the first in line among both adult outpatients and inpatients. During the study period, a PIV-3 outbreak (n= 96) have emerged between December 2004-April 2005, and an influenza A (H1N1)pdm09 outbreak (n= 207) between November 2009-January 2010. When the seasonal distribution was considered, the isolation rates of 1705 respiratory viruses in winter, spring, summer and autumn were 44.4%, 27%, 8.3% and 20.3%, respectively. RSV was most frequently detected from December to March, influenza viruses from November to March, HRV from December to June, and mixed infections from January to February. In conclusion, the data of our study obtained in about 12-year period indicated that the prevalence of respiratory viruses in acute respiratory infections is 33.4%, and they typically active during the months of winter and early spring in our region.


title: Human coronavirus 229E: receptor binding domain and neutralization by soluble receptor at 37 degrees C.

text: Truncated human coronavirus HCoV-229E spike glycoproteins containing amino acids 407 to 547 bound to purified, soluble virus receptor, human aminopeptidase N (hAPN). Soluble hAPN neutralized the infectivity of HCoV-229E virions at 37 degrees C, but not 4 degrees C. Binding of hAPN may therefore trigger conformational changes in the viral spike protein at 37 degrees C that facilitate virus entry.


title: Diversity of influenza-like illness etiology in Polish Armed Forces in influenza epidemic season.

text: The aim of this study was to conduct an epidemiological and laboratory surveillance of Influenza-Like Illnesses (ILI) in Polish Armed Forces, civilian military personnel and their families in 2011/2012 epidemic season, under the United States Department of Defense-Global Emerging Infections Surveillance and Response System (DoD-GEIS). ILI incidence data were analyzed in relation to age, gender, patient category as well as pathogen patterns. Multiple viral, bacterial and viral-bacterial co-infections were identified. Nose and throat swabs of active duty soldiers in the homeland country and in the NATO peacekeeping forces KFOR (Kosovo Force), as well as members of their families were tested for the presence of viral and bacterial pathogens. From October 2011 to May 2012, 416 specimens from ILI symptoms patients were collected and analyzed for the presence of viral and bacterial pathogens. Among viruses, coronavirus was the most commonly detected. In the case of bacterial infections, the most common pathogen was Staphylococcus aureus.


title: Bladder cancer. Clinical practice guidelines in oncology.

text: Urothelial tumors represent a spectrum of diseases with a range of prognosis. After a diagnosis is established at any point within the urothelial tract, the patient remains at risk for developing a new lesion at a different or the same location and at a similar or more advanced stage. Continued monitoring for recurrence is an essential part of management, because most recurrences are superficial and can be managed endoscopically. Within each category of disease, more refined methods to determine prognosis and guide management, based on molecular staging, are under development with the goal of optimizing the individual patient's likelihood of cure and chance for organ preservation. For patients with more extensive disease, newer treatments typically involve combined-modality approaches, using recently developed surgical procedures, or three-dimensional treatment planning for more precise delivery of radiation therapy. Although these are not appropriate in all cases, they do offer the promise of an improved quality of life and prolonged survival. Finally, within the category of metastatic disease, a number of new agents have been identified that appear to be superior to those currently considered to be standard therapies. Experts believe, therefore, that the treatment of urothelial tumors will evolve rapidly over the next few years, with improved outcomes for patients at all stages of disease.


title: Development and evaluation of reverse transcription-loop-mediated isothermal amplification assay for rapid and real-time detection of Japanese encephalitis virus.

text: The standardization and validation of a one-step, single-tube accelerated quantitative reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay is reported for rapid and real-time detection of Japanese encephalitis virus (JEV). The RT-LAMP assay reported in this study is very simple and rapid; the amplification can be obtained in 30 min under isothermal conditions at 63 degrees C by employing a set of six primers targeting the E gene of JEV. The RT-LAMP assay demonstrated exceptionally higher sensitivity compared to that of RT-PCR, with a detection limit of 0.1 PFU. The specificities of the selected primer sets were established by cross-reactivity studies with other closely related members of the JEV serocomplex as well as by evaluation of healthy human volunteers. The comparative evaluation of the RT-LAMP assay for clinical diagnosis with a limited number of patient cerebrospinal fluid samples revealed 85% concordance with conventional RT-PCR, with a sensitivity and a specificity of 100% and 86%, respectively. The concentration of virus in most of the clinical samples was 10(2) to 10(5) PFU/ml, as determined from the standard curve based on the time of positivity in the samples. In addition, the monitoring of gene amplification can also be visualized with the naked eye by using SYBR green I fluorescent dye. Thus, due to easy operation without a requirement of sophisticated equipment and skilled personnel, the RT-LAMP assay reported here is a valuable tool for the rapid and real-time detection of JEV not only by well-equipped laboratories but also by peripheral diagnostic laboratories with limited financial resources in developing countries.


title: Qigong practice among chronically ill patients during the SARS outbreak.

text: AIMS AND OBJECTIVES This research aimed at exploring the motivations and experiences of chronically ill participants practising qigong during the severe acute respiratory syndrome outbreak in Hong Kong. BACKGROUND Although biomedicine is the mainstream medical system in Hong Kong, many people employ complementary and alternative medicine in dealing with their chronic health problems. Practising qigong is one of the most popular forms of complementary and alternative medicine used among chronically ill patients. Little is known about the experiences of the chronically ill patients practising qigong in the severe acute respiratory syndrome outbreak and even less is known how this practice is related to the social context of Hong Kong during the outbreak. DESIGN AND METHODS Qualitative methods using participant-observation from three qigong classes in Hong Kong and in-depth semi-structured interviews with 30 participants were employed. Content analysis and discourse analysis were used to identify major themes of the data. RESULTS Both the underlying and trigger motivations could motivate these chronically ill participants to practise qigong. Legitimacy of qigong in health maintenance, deterioration of health and unpleasant experiences in biomedical treatment on their chronic illnesses served as the underlying motivations. Stigmatization of and discrimination against the chronically ill and the sense of searching coping strategy during the severe acute respiratory syndrome outbreak further motivated their practice. CONCLUSION To the participants, practising qigong not only could strengthen their health, but was also a coping strategy for them to regain an active control and the sense of security in their health and enabled them to overcome the social stigmatization and discrimination during the outbreak. RELEVANCE TO CLINICAL PRACTICE. Searching for emotional support and a more active role in health conditions could be seen from the chronically ill participants. Understanding and paying more attention to the particular needs of the chronically ill patients can enable health-care professionals to provide better care and support for the chronically ill during an epidemic crisis.


title: Construction of a mouse hepatitis virus recombinant expressing a foreign gene.

text: The genome of the coronavirus mouse hepatitis virus (MHV) contains genes which have been shown to be nonessential for viral replication and which could, in principle, be used as sites for the introduction of foreign sequences. We have inserted heterologous genetic material into gene 4 of MHV in order (i) to test the applicability of targeted RNA recombination for site-directed mutagenesis of the MHV genome upstream of the N gene; (ii) to develop further genetic tools for mutagenesis of structural genes other than N; and (iii) to examine the feasibility of using MHV as an expression vector. A DI-like donor RNA vector containing the MHV S gene and all genes distal to S was constructed. Initially, a derivative of this was used to insert a 19-nucleotide tag into the start of ORF 4a of MHV-A59 using the N gene deletion mutant A1b4 as the recipient virus. Subsequently, the entire gene for the green fluorescent protein (GFP) was inserted in place of gene 4. This heterologous gene was shown to be expressed by recombinant viruses but not at levels sufficient to allow detection of fluorescence of viral plaques. Northern blot analysis of transcripts of GFP recombinants showed the expected displacement of the mobility, relative to those of wild-type, of all subgenomic mRNAs larger than mRNA5. An unexpected result of the Northern analysis was the observation that GFP recombinants also produced an RNA species the same size as that of wild-type mRNA4. RT-PCR analysis of the 5' end of this species revealed that it was actually a collection of mRNAs originating from a cluster of 10 different sites, none of which possessed a canonical intergenic sequence. The finding of these aberrant mRNAs, all of nearly the same size as wild-type mRNA4, suggests that long range structure of the MHV genome can sometimes be the sole determinant of the site of initiation of transcription.


DNA vaccines and apoptosis: to kill or not to kill?


docid: bwwgxeeg

title: Supplementation with 3 compositionally different tocotrienol supplements does not improve cardiovascular disease risk factors in men and women with hypercholesterolemia.

text: BACKGROUND Tocotrienols have been reported to lower LDL-cholesterol and fasting glucose concentrations and to have potent antioxidant effects, but the results are contradictory. OBJECTIVE The objective was to study the relative effect of tocotrienol supplements of different compositions (mixed alpha- plus gamma-, high gamma-, or P25-complex tocotrienol) on blood lipids, fasting blood glucose, and the excretion of 8-iso-prostaglandin F(2alpha), a measure of oxidative stress, in healthy hypercholesterolemic men and women. DESIGN This was a double-blind, randomized, parallel-design study in which subjects (n = 67 men and women) consumed 1 of 3 commercially available tocotrienol supplements or a safflower oil placebo for 28 d. Blood and urine samples were obtained before and after the 28-d supplementation phase for analysis of fasting blood lipids, glucose, tocotrienols and tocopherols, and 8-iso-prostaglandin F(2alpha). RESULTS Overall, serum tocotrienols were increased in subjects who consumed tocotrienols, which showed that the putatively active components were absorbed. No significant differences in mean lipid or glucose concentrations were observed among the 4 treatment groups at the end of the 28-d supplementation phase. However, when the values were expressed as a percentage change from the concentrations during the presupplementation run-in phase, LDL cholesterol increased slightly (7 +/- 2%) but significantly (P < 0.05) in the group consuming the mixed alpha- plus gamma-tocotrienol supplement when compared with LDL cholesterol in the group consuming the P25-complex tocotrienol. Neither mean concentrations nor the percentage change in 8-iso-prostaglandin F(2alpha) differed significantly among treatments. CONCLUSION Supplementation with 200 mg tocotrienols/d from 3 commercially available sources has no beneficial effect on key cardiovascular disease risk factors in highly compliant adults with elevated blood lipid concentrations.


title: Multimeric humanized varicella-zoster virus antibody fragments to gH neutralize virus while monomeric fragments do not.

text: Murine monoclonal antibody 206 (MAb mu206) binds to gH, the varicella-zoster virus (VZV) fusogen, neutralizing the virus in vitro in the absence of complement and inhibiting cell-to-cell spread and egress of VZV in cultured cells. We have humanized this antibody to generate MAb hu206 by complementarity determining region grafting. MAb hu206 retained binding and in vitro neutralizing activity, as well as cross-reactivity with ten different VZV strains. Single-chain antibody fragments (scAb) derived from MAb hu206 were produced in Escherichia coli. These scAb retained the binding properties of the whole antibody. However, monomeric scAb exhibited markedly reduced neutralizing activity compared to the bivalent parental MAb hu206. Shortening the peptide linker joining the V(H) to the V(kappa) domain from 14 to 5 or even 0 residues encouraged multimerization and increased neutralizing efficacy. The fact that Fab fragments enzymatically generated from whole MAb hu206 lost their neutralizing potency lent support to the proposal that valency is important for VZV neutralization at this epitope.


title: Molecular detection and identification of influenza viruses by oligonucleotide microarray hybridization.

text: Microarrays of virus-specific oligonucleotides may provide a method of screening samples for the presence or absence of a large variety of viruses simultaneously. Influenza viruses are ideal for evaluating such microarrays because of their genetic and host diversity, and the availability of an extensive sequence database. A collection of 476 influenza virus-specific oligonucleotides was spotted onto glass slides as probes. Viral RNAs were reverse transcribed and amplified by PCR, and the products were labeled with cyanine dyes. The presence of viruses and their identities were determined by hybridization. The fluorescence intensities of oligonucleotide spots were highly reproducible within each slide and satisfactorily proportional between experiments. However, the intensities of probe spots completely complementary to target sequences varied from background to saturation. The variations did not correlate with base composition, nucleotide sequence, or internal secondary structures. Therefore, thresholds for determining whether hybridization to a spot should be judged as positive were assigned individually. Considering only positive spots from probes predicted to be monospecific for influenza virus species, subtype, host source, or gene segment, this method made correct identifications at the species, hemagglutinin subtype, and gene segment levels. Monospecific neuraminidase (NA) subtype probes were insufficiently diverse to allow confident NA subtype assignment. Incorporating positive spots from polyspecific probes into the identification scheme gave similar results. Overall, the results demonstrate the potential of microarray-based oligonucleotide hybridization for multiple virus detection.


docid: rdywyxqz

title: Hendra and nipah infection: pathology, models and potential therapies.

text: The Paramyxoviridae family comprises of several genera that contain emerging or re-emerging threats for human and animal health with no real specific effective treatment available. Hendra and Nipah virus are members of a newly identified genus of emerging paramyxoviruses, Henipavirus. Since their discovery in the 1990s, henipaviruses outbreaks have been associated with high economic and public health threat potential. When compared to other paramyxoviruses, henipaviruses appear to have unique characteristics. Henipaviruses are zoonotic paramyxoviruses with a broader tropism than most other paramyxoviruses, and can cause severe acute encephalitis with unique features among viral encephalitides. There are currently no approved effective prophylactic or therapeutic treatments for henipavirus infections. Although ribavirin was empirically used and seemed beneficial during the biggest outbreak caused by one of these viruses, the Nipah virus, its efficacy is disputed in light of its lack of efficacy in several animal models of henipavirus infection. Nevertheless, because of its highly pathogenic nature, much effort has been spent in developing anti-henipavirus therapeutics. In this review we describe the unique features of henipavirus infections and the different strategies and animal models that have been developed so far in order to identify and test potential drugs to prevent or treat henipavirus infections. Some of these components have the potential to be broad-spectrum antivirals as they target effectors of viral pathogenecity common to other viruses. We will focus on small molecules or biologics, rather than vaccine strategies, that have been developed as anti-henipaviral therapeutics.


Immunopathogenesis of demyelination induced by MHV-4.


Orogastric administration of crushed darunavir tablets for a critically ill patient.


Saint Louis University School of Medicine and Vaccine Center Mobilize for COVID-19 Pandemic.


Using BRCA1 to treat cancer.


docid: bymwqehy

title: A cross-sectional validation study of using NERDS and STONEES to assess bacterial burden.

text: All chronic wounds are colonized by micro-organisms. Although the presence of bacteria is not necessarily harmful, and may be beneficial in some instances, accurate evaluation of wound-related bacterial damage and infection is crucial. A cross-sectional validation study involving 112 patients was conducted to estimate the specificity and sensitivity of clinical assessment variables individually and in combination to determine the presence and quantity of bacteria in the wound. The average age of study participants was 66 years (range 33 to 95 years) and most had leg (44) and foot (68) ulcers of approximately 6 months' duration. Wounds were evaluated using a mnemonic developed to evaluate the presence or absence of clinical signs of critical colonization (NERDS) or infection (STONEES) and results compared to semi-quantitative swab cultures. Wounds with debris, increased exudate, and friable tissue were found to be five times more likely to have scant or light bacterial growth; whereas, wounds with elevated temperature were eight times more likely to have moderate or heavy bacterial growth. When combining any three clinical signs, the sensitivity was 73.3% for scant or light and 90% for moderate and heavy bacterial growth and the specificity was 80.5% and 69.4%, respectively. Considering the importance of this clinical diagnosis, studies to examine the predictive validity of these assessment variables and culture results are warranted.


title: Rat strains differ in susceptibility to Ureaplasma parvum-induced urinary tract infection and struvite stone formation.

text: Individuals with struvite uroliths are susceptible to recurrent urinary tract infections (UTI), sepsis, and renal disease. Unfortunately, little is known about the host-specific factors that predispose to this disease. In order to develop a rodent model that can address this problem, we inoculated female Fischer 344 (F344), Lewis (LEW), Sprague-Dawley (SD), and Wistar (WIS) rats with a host-adapted strain of Ureaplasma parvum. Animals were necropsied at 2 weeks postinoculation; 100% of F344, 42% of SD, 10% of LEW, and 10% of WIS rats remained infected. Severe bladder lesions and struvite calculi were seen in 64% of F344 rats; in other rat strains, bladder lesions were mild or absent. F344 rats with struvite uroliths had the highest urinary levels of proinflammatory cytokines, such as GRO/KC, interleukin-1alpha (IL-1alpha), and IL-1beta. F344 rats without struvite stones at necropsy had milder bladder lesions and significantly lower urinary levels of proinflammatory cytokines but a more prominent inflammatory response than did other rat strains. Based on our results, struvite stone formation is linked to a robust inflammatory response that does not resolve UTI but instead promotes damage to surrounding tissues.


title: mRNA helicase activity of the ribosome.

text: Most mRNAs contain secondary structure, yet their codons must be in single-stranded form to be translated. Until now, no helicase activity has been identified which could account for the ability of ribosomes to translate through downstream mRNA secondary structure. Using an oligonucleotide displacement assay, together with a stepwise in vitro translation system made up of purified components, we show that ribosomes are able to disrupt downstream helices, including a perfect 27 base pair helix of predicted T(m) = 70 degrees . Using helices of different lengths and registers, the helicase active site can be localized to the middle of the downstream tunnel, between the head and shoulder of the 30S subunit. Mutation of residues in proteins S3 and S4 that line the entry to the tunnel impairs helicase activity. We conclude that the ribosome itself is an mRNA helicase and that proteins S3 and S4 may play a role in its processivity.


title: [Detection of the mRNA expression of human angiotensin-converting enzyme 2 as a SARS coronavirus functional receptor in human femoral head].

text: OBJECTIVE To investigate the mRNA expression of severe acute respiratory syndrome-associated coronavirus (SARS-COV) functional receptor, angiotensin-converting enzyme 2 (ACE2), in human femoral head and conjunctiva, and explore the possible entry route of SARS-COV in human femoral head. METHODS ACE2 mRNA in human femoral head was detected by nested RT-PCR with human beta actin gene as the positive control. RESULTS The mRNA of human beta actin gene could be amplified efficiently in all the tissue samples. The mRNA of human ACE2 was expressed efficiently in the normal lung tissue, but not in the cartilage and cancellous bone under the weight-bearing area of the femoral head. CONCLUSION SARS-COV can not infect the femoral head tissue and lead to avascular necrosis of the femoral head directly by the spike glycoprotein, and mechanism of the virus for causing avascular necrosis needs further investigation.


title: Lymphopenia in severe acute respiratory syndrome: a summary on its frequency.

text: Severe acute respiratory syndrome (SARS) is a new human infectious disease caused by coronavirus. The first world outbreak occurs in 2002, starting from China and then thorough many countries. Abnormal hematological variables were common among patients with SARS. Concerning lymphocyte disorder, there were some previous studies reported the low lymphocyte in the patients with SARS. Here, the author performs a further analysis to document the frequency of the lymphopenia in SARS by the way of metanalysis study. Five available reports concerning the prevalence of lymphopenia in SARS among different populations were selected for further analysis in this study. Overall 637 SARS patients were retrospectively analyzed and 492 cases presented lymphopenia. According to this study, the overall prevalence rate of lymphopenia in SARS is 77.2 %. No significant correlation between the population ethnicity and the prevalence rate was detected in this study (P > 0.05). Here, the lymphopenia can be documented as an important characteristic of blood picture in the patients with SARS or it can indicate that "SARS is a viral-induced lymphopenia disease".


title: A panel of optimized primers and positive-control DNAs for PCR detection of common biological contaminants in mouse cell lines and tissue samples.

text: PCR-based testing for infectious agents in mouse cell lines and tissues has recently been developed as an alternative to the traditional MAP test. One drawback to currently available PCR-based assays is the lack of appropriate positive controls for PCR detection of the infectious agents. When negative samples are the norm and positive controls are absent, it is very difficult to feel confident detecting infectious agents. To alleviate this problem, the authors developed a panel of primers and positive-control DNA plasmids that enable rapid testing of biological samples, such as cell lines, tissues, or animal sera, for presence of the infectious agents most damaging to mouse colonies.


docid: rtsllgsr

title: Intracellular complexes of viral spike and cellular receptor accumulate during cytopathic murine coronavirus infections.

text: Murine hepatitis virus (MHV) infections exhibit remarkable variability in cytopathology, ranging from acutely cytolytic to essentially asymptomatic levels. In this report, we assess the role of the MHV receptor (MHVR) in controlling this variable virus-induced cytopathology. We developed human (HeLa) cell lines in which the MHVR was produced in a regulated fashion by placing MHVR cDNA under the control of an inducible promoter. Depending on the extent of induction, MHVR levels ranged from less than approximately 1,500 molecules per cell (designated R(lo)) to approximately 300,000 molecules per cell (designated R(hi)). Throughout this range, the otherwise MHV-resistant HeLa cells were rendered susceptible to infection. However, infection in the R(lo) cells occurred without any overt evidence of cytopathology, while the corresponding R(hi) cells died within 14 h after infection. When the HeLa-MHVR cells were infected with vaccinia virus recombinants encoding MHV spike (S) proteins, the R(hi) cells succumbed within 12 h postinfection; R(lo) cells infected in parallel were intact, as judged by trypan blue exclusion. This acute cytopathology was not due solely to syncytium formation between the cells producing S and MHVR, because fusion-blocking antiviral antibodies did not prevent it. These findings raised the possibility of an intracellular interaction between S and MHVR in the acute cell death. Indeed, we identified intracellular complexes of S and MHVR via coimmunoprecipitation of endoglycosidase H-sensitive forms of the two proteins. We suggest that MHV infections can become acutely cytopathic once these intracellular complexes rise above a critical threshold level.


Hormone receptors in breast cancer.


title: Noninvasive positive pressure ventilation in the intensive care unit: a concise review.

text: OBJECTIVE To critically assess available high-level clinical studies regarding use of noninvasive positive pressure ventilation in varied intensive care unit settings. DATA SOURCE Search of pertinent articles within Ovid MEDLINE from 1975 to 2005, CINAHL from 1982 to 2005, EMBASE from 1988 to 2005, and Web of Science from 1993 to 2005. STUDY SELECTION Randomized, controlled clinical trials and cohort studies and observational studies the authors consider important or novel. DATA EXTRACTION/SYNTHESIS Performed equally by both authors with the use of an Excel data spreadsheet. CONCLUSION There is abundant level I evidence supporting the use of noninvasive positive pressure ventilation in such critical care settings as acute hypercapnic respiratory failure, particularly related to chronic obstructive pulmonary disease, and acute cardiogenic pulmonary edema. We also report on other clinical scenarios in which the data may be somewhat less compelling, but evidence favors a noninvasive positive pressure ventilation trial. Some well designed studies suggest that noninvasive positive pressure ventilation is not an appropriate intervention for patients who have failed endotracheal extubation.


Rechallenging immunological memory.


title: The association of titers to bovine coronavirus with treatment for bovine respiratory disease and weight gain in feedlot calves.

text: The association between bovine respiratory disease (BRD) and antibody titers to bovine coronavirus (BCV) was studied in 604 calves (19 different groups in 4 different feedlots from 2 provinces). Almost all calves had antibody titers on arrival in the Alberta feedlot and 82% of the calves had an antibody titer on arrival at the Ontario feedlots; titers in calves in Alberta were almost twice as high as those in calves in Ontario. The incidence of infection, in the first mo after arrival as judged by seroconversion, ranged from 61% to 100%; titer increases were much greater in calves in Ontario feedlots. Titer variables were not significantly related to BRD, except on a within-group basis (group was a confounding variable for BCV-BRD associations). Given control of group effects, calves with an antibody titer on arrival appeared to be protected against BRD for the first 28 d in the feedlot, and the association was reasonably linear over the range of titers. Each titer unit on arrival decreased the risk of BRD by about 0.8x (odds ratio). Titer change was not strongly related to the risk of BRD and the relationship was not linear over the range of titer changes. Titer change was strongly and negatively correlated with titer on arrival, and titer change was not significantly related to BRD in the presence of arrival titers. Arrival titer retained its relationship with BRD in the presence of titer data for other putative pathogens. Each higher unit of titer to BCV on arrival increased the 28-day weight gain (controlling for group, initial weight and the occurrence of BRD) by slightly more than 1 kg. Titer change was associated with decreased weight gain, when initial titer was not in the model. The lack of a linear or multivariable association between BCV titer change and BRD, and weight gain, may indicate that BCV is not a major pathogen; or, its lack of significance may merely be due to its strong correlation with arrival titer. Given the associations found in this study, particularly the interprovincial differences in arrival titers, more and different approaches to studying the possible effects of BCV on BRD are in order.


Planetary health: what is it and what should doctors do?


title: Expression of reporter genes from the defective RNA CD-61 of the coronavirus infectious bronchitis virus.

text: The defective RNA (D-RNA) CD-61, derived from the Beaudette strain of the avian coronavirus infectious bronchitis virus (IBV), was used as an RNA vector for the expression of two reporter genes, luciferase and chloramphenicol acetyltransferase (CAT). D-RNAs expressing the CAT gene were demonstrated to be capable of producing CAT protein in a helper-dependent expression system to about 1.6 microgram per 10(6) cells. The reporter genes were expressed from two different sites within the CD-61 sequence and expression was not affected by interruption of the CD-61-specific ORF. Expression of the reporter genes was under the control of a transcription-associated sequence (TAS) derived from the Beaudette gene 5, normally used for the transcription of IBV subgenomic mRNA 5. The Beaudette gene 5 TAS is composed of two tandem repeats of the IBV canonical consensus sequence involved in the acquisition of a leader sequence during the discontinuous transcription of IBV subgenomic mRNAs. It is demonstrated that only one canonical sequence is required for expression of mRNA 5 or for the expression of an mRNA from a D-RNA and that either sequence can function as an acceptor site for acquisition of the leader sequence.


title: Lyme disease--United States, 2003-2005.

text: Lyme disease is caused by the spirochete Borrelia burgdorferi and is transmitted to humans by the bite of infected blacklegged ticks (Ixodes spp.). Early manifestations of infection include fever, headache, fatigue, and a characteristic skin rash called erythema migrans. Left untreated, late manifestations involving the joints, heart, and nervous system can occur. A Healthy People 2010 objective (14-8) is to reduce the annual incidence of Lyme disease to 9.7 new cases per 100,000 population in 10 reference states where the disease is endemic (Connecticut, Delaware, Maryland, Massachusetts, Minnesota, New Jersey, New York, Pennsylvania, Rhode Island, and Wisconsin). This report summarizes surveillance data for 64,382 Lyme disease cases reported to CDC during 2003-2005, of which 59,770 cases (93%) were reported from the 10 reference states. The average annual rate in these 10 reference states for the 3-year period (29.2 cases per 100,000 population) was approximately three times the Healthy People 2010 target. Persons living in Lyme disease--endemic areas can take steps to reduce their risk for infection, including daily self-examination for ticks, selective use of acaricides and tick repellents, use of landscaping practices that reduce tick populations in yards and play areas, and avoidance of tick-infested areas.


title: Serosurvey of viral infections in free-ranging Namibian cheetahs (Acinonyx jubatus).

text: Cheetahs (Acinonyx jubatus) in captivity have unusually high morbidity and mortality from infectious diseases, a trait that could be an outcome of population homogeneity or the immunomodulating effects of chronic stress. Free-ranging Namibian cheetahs share ancestry with captive cheetahs, but their susceptibility to infectious diseases has not been investigated. The largest remaining population of free-ranging cheetahs resides on Namibian farmlands, where they share habitat with domestic dogs and cats known to carry viruses that affect cheetah health. To assess the extent to which free-ranging cheetahs are exposed to feline and canine viruses, sera from 81 free-ranging cheetahs sampled between 1992 and 1998 were evaluated for antibodies against canine distemper virus (CDV), feline coronavirus (feline infectious peritonitis virus; FCoV/ FIPV), feline herpesvirus 1 (FHV1), feline panleukopenia virus (FPV), feline immunodeficiency virus (FIV), and feline calicivirus (FCV) and for feline leukemia virus (FeLV) antigens. Antibodies against CDV, FCoV/FIPV, FHV1, FPV, and FCV were detected in 24, 29, 12, 48, and 65% of the free-ranging population, respectively, although no evidence of viral disease was present in any animal at the time of sample collection. Neither FIV antibodies nor FeLV antigens were present in any free-ranging cheetah tested. Temporal variation in FCoV/FIPV seroprevalence during the study period suggested that this virus is not endemic in the free-ranging population. Antibodies against CDV were detected in cheetahs of all ages sampled between 1995 and 1998, suggesting the occurrence of an epidemic in Namibia during the time when CDV swept through other parts of sub-Saharan Africa. This evidence in free-ranging Namibian cheetahs of exposure to viruses that cause severe disease in captive cheetahs should direct future guidelines for translocations, including quarantine of seropositive cheetahs and preventing contact between cheetahs and domestic pets.


title: Could SARS-CoV-2/COVID-19 simply fade away?

text: not available.


title: The complete genome sequence of the major component of a mild citrus tristeza virus isolate.

text: The genome of the Spanish mild isolate T385 of citrus tristeza virus (CTV) was completely sequenced and compared with the genomes of the severe isolates T36 (Florida), VT (Israel) and SY568 (California). The genome of T385 was 19,259 nt in length, 37 nt shorter than the genome of T36, and 33 and 10 nt longer than those of VT and SY568, respectively, but their organization was identical. T385 had mean nucleotide identities of 81.3, 89.3 and 94% with T36, VT and SY568, respectively. The 3' UTR had over 97% identity in all isolates, whereas the 5' UTR of T385 had 67% identity with VT, 66.3% with SY568 and only 42.5% with T36. In the coding regions, the nucleotide differences between T385 and VT were evenly distributed along the genome (around 90% identity); this was not observed between T385 and the other isolates. T385 and T36 had nucleotide identities around 90% in the eight 3'-terminal ORFs of the genome, but only 72.3% in ORF 1a, a divergence pattern similar to that reported previously for T36 and VT. T385 and SY568 had nucleotide identities close to 90% in the 5'- and 3'-terminal regions of the genome, whereas the central region had over 99% identity. Our data suggest that the central region in the SY568 genome results from RNA recombination between two CTV genomes, one of which was almost identical to T385.


title: Genetic determinants of Sindbis virus mosquito infection are associated with a highly conserved alphavirus and flavivirus envelope sequence.

text: Wild-type Sindbis virus (SINV) strain MRE16 efficiently infects Aedes aegypti midgut epithelial cells (MEC), but laboratory-derived neurovirulent SINV strain TE/5'2J infects MEC poorly. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of MRE16 and TE/5'2J differ at 60 residue sites. To identify the genetic determinants of MEC infection of MRE16, the TE/5'2J virus genome was altered to contain either domain chimeras or more focused nucleotide substitutions of MRE16. The growth patterns of derived viruses in cell culture were determined, as were the midgut infection rates (MIR) in A. aegypti mosquitoes. The results showed that substitutions of MRE16 E2 aa 95 to 96 and 116 to 119 into the TE/5'2J virus increased MIR both independently and in combination with each other. In addition, a unique PPF/.GDS amino acid motif was located between these two sites that was found to be a highly conserved sequence among alphaviruses and flaviviruses but not other arboviruses.


title: Human rhinovirus infection in children with cystic fibrosis.

text: Nasopharyngeal swabs obtained from 78 pediatric patients with cystic fibrosis (CF), including 47 with acute pulmonary exacerbation and 31 in a stable clinical condition, were evaluated for 17 respiratory viruses. Human rhinovirus (HRV) was the most frequently detected virus in patients with pulmonary exacerbation and in those who were clinically stable (21.3% vs. 12.9%; P = 0.52). HRV-A was the main RV detected in patients with pulmonary exacerbations. However, no prevalence of particular HRV-A subtypes was found. This study highlights that RV is frequently found in the respiratory secretions of patients with CF and the impact of HRV-A appears higher than that of the other HRV types in patients with pulmonary exacerbations.


title: Differences among confirmed and not-confirmed COVID-19 patients at "D.Cotugno" hospital, Naples (Italy): what we learned from first suspected cases?

text: Clinical presentation of COVID-19 is common to other respiratory infections. We compared the characteristics at hospital admission of confirmed and not-confirmed COVID-19 patients, in the early phase of the epidemic. Thirty-seven suspected patients were enrolled, and COVID-19 was confirmed in 17. Confirmed patients are older, have more frequently contact with confirmed cases. Distinctive clinical characteristics among COVID-19 were the grand-glass opacities at CT scan, and a pO2/FiO2 ratio less than 250. In not-confirmed group, Influenza represented the most frequent alternative diagnosis. This study contributes to highlight the characteristics to consider at hospital admission in order to promptly suspect COVID-19.


Review Team.


title: Epidemiological study of enteric coronavirus excretion by an enzyme-linked immunosorbent assay.

text: The enzyme-linked immunosorbent assay with a blocking test was used to determine the prevalence of enteric coronavirus excretion among the population of Bangkok and Khon Kaen province. The results indicated that coronaviruses were present in fecal samples from both children and adults with or without diarrhea. It is interesting that the viruses were more frequently observed in the apparently healthy individuals in Bangkok than in the patients with diarrhea whereas the excretion rates were approximately alike in both groups in Khon Kaen province. Among children aged less than 5 years (with or without diarrhea) the viruses were more frequently observed in stools from young children (1-5 years old) than from the newborn (less than 1-year-old) particularly those living in the Bang Khen slum area and in the rural Khon Kaen community. The rate of coronavirus excretion did not increase with age and some apparently healthy individuals continued to excrete the viruses for several months. The results also suggested that chronic parasitic infection associated with low socioeconomic status might influence the excretion of enteric coronaviruses in the feces.


The resurgence of West Nile virus.


title: [SARS: diagnosis, therapy, and especially prevention].

text: The main purpose of this review is to analyze some aspects of the severe acute respiratory syndrome, SARS, in order to obtain useful data to suggest preventive actions to reduce the spreading of the disease. Many elements have been examined to reach some conclusions and to allow an updated discussion. Surgical masks protect more the patient than the caregiver. Simple or double surgical masks may be useful, as double gloving protects the hands of the surgical personnel against percutaneous transmission of HIV eventually present in contaminated blood. The frequent substitution of the external masks with a new one will improve the filtering activity against droplets produced by cough or sneezes of the patient. The use of respiratory masks may be suggested in hospitals or in restricted ventilated areas where, even if coronavirus variant is considered an environmental contaminant more than a respiratory risk, droplets nuclei may persist in the air and add consistent dangers to the heath-care givers. Considering that large and medium droplets may infect floors and surfaces, in addition to gloves, gowns, masks and eyes protection, the available list of viral and bacterial factors implicated in SARS ethiology suggests a better hand antisepsis using frequently the alcohol based gels (containing an high percentage of emollients substances), if available. A liquid soap with triclosan can also be used, if the health-care workers compliance to hand washing increases, as expected in this explosive situation. On the basis of the results of some experimental data, the environmental disinfection may be effected with ethyl alcohol 70% in water. Disinfection of floors or larger surfaces may be obtained with chlorine compounds solutions, after an accurate pre-cleaning. When corrosion, bleaching or gas production have to be avoided, chlorine compounds may be substituted by phenolic detergent disinfectants.