title: The indirect fluorescent antibody technique as a method for detecting antibodies in aborted fetuses.

text: In this investigation the indirect fluorescent antibody technique was used to titrate antibodies in bovine sera to parainfluenza 3, infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. These results were compared to those determined on the same samples by hemagglutination inhibition for parainfluenza 3 virus and serum neutralization for bovine virus diarrhea and infectious bovine rhinotracheitis virus. The results of the serological methods agreed closely. The indirect fluorescent antibody technique is a rapid and sensitive method for detecting antibodies and the procedure lends itself to use in diagnostic laboratories. In addition to the above viruses the presence or absence of antibodies to bovine coronavirus and bovine adenovirus 3 were determined by the indirect fluorescent antibody technique in thoracic fluids from 100 aborted fetuses and 50 nonaborted fetuses. Results on these samples were not compared to hemagglutination inhibition or serum neutralization as the condition of fluid samples from aborted fetuses renders interpretation of such tests unreliable. Antibodies to one or more viruses were detected in 30 of the 100 aborted fetuses and in seven of the 50 nonaborted fetuses. Antibodies to more than one agent were detected in eleven of the 100 aborted and in one of the 50 nonaborted fetuses. Reasons for this occurrence and application of the test in determination of causes of abortion are discussed.


title: Xenotransplantation: an overview of microbiological risks and potentials for risk management.

text: Xenotransplantation is the use of animal organs, tissues or cells for transplantion into humans to treat a variety of medical conditions. If proven efficacious, the technique could be used as one means of alleviating the disparity between the growing demand for transplantable organs, tissues and cells, and the availability of human-origin transplants world-wide. Just as the practicality and efficacy of the technology need to be investigated, so too does the potential for associated infectious disease risk. While much remains to be learned about the microbiological risk associated with xenotransplantation, the elements to be incorporated into xenotransplantation risk management schemes can be considered, using what is currently known about the infectious agents potentially relevant to the xenotransplantation setting.


title: Certolizumab pegol for the treatment of Crohn's disease.

text: BACKGROUND Certolizumab pegol is a pegylated humanized Fab' fragment that binds tumor necrosis factor alpha. METHODS In a randomized, double-blind, placebo-controlled trial, we evaluated the efficacy of certolizumab pegol in 662 adults with moderate-to-severe Crohn's disease. Patients were stratified according to baseline levels of C-reactive protein (CRP) and were randomly assigned to receive either 400 mg of certolizumab pegol or placebo subcutaneously at weeks 0, 2, and 4 and then every 4 weeks. Primary end points were the induction of a response at week 6 and a response at both weeks 6 and 26. RESULTS Among patients with a baseline CRP level of at least 10 mg per liter, 37% of patients in the certolizumab group had a response at week 6, as compared with 26% in the placebo group (P=0.04). At both weeks 6 and 26, the corresponding values were 22% and 12%, respectively (P=0.05). In the overall population, response rates at week 6 were 35% in the certolizumab group and 27% in the placebo group (P=0.02); at both weeks 6 and 26, the response rates were 23% and 16%, respectively (P=0.02). At weeks 6 and 26, the rates of remission in the two groups did not differ significantly (P=0.17). Serious adverse events were reported in 10% of patients in the certolizumab group and 7% of those in the placebo group; serious infections were reported in 2% and less than 1%, respectively. In the certolizumab group, antibodies to the drug developed in 8% of patients, and antinuclear antibodies developed in 2%. CONCLUSIONS In patients with moderate-to-severe Crohn's disease, induction and maintenance therapy with certolizumab pegol was associated with a modest improvement in response rates, as compared with placebo, but with no significant improvement in remission rates. (ClinicalTrials.gov number, NCT00152490 [ClinicalTrials.gov].).


On the risk of introduction of an emerging infectious disease.


title: Contribution of the intercalated adenosine at the helical junction to the stability of the gag-pro frameshifting pseudoknot from mouse mammary tumor virus.

text: The mouse mammary tumor virus (MMTV) gag-pro frameshifting pseudoknot is an H-type RNA pseudoknot that contains an unpaired adenosine (A14) at the junction of the two helical stems required for efficient frameshifting activity. The thermodynamics of folding of the MMTV vpk pseudoknot have been compared with a structurally homologous mutant RNA containing a G x U to G-C substitution at the helical junction (U13C RNA), and an A14 deletion mutation in that context (U13CdeltaA14 RNA). Dual wavelength optical melting and differential scanning calorimetry reveal that the unpaired adenosine contributes 0.7 (+/-0.2) kcal mol(-1) at low salt and 1.4 (+/-0.2) kcal mol(-1) to the stability (deltaG(0)37) at 1 M NaCl. This stability increment derives from a favorable enthalpy contribution to the stability deltadeltaH = 6.6 (+/-2.1) kcal mol(-1) with deltadeltaG(0)37 comparable to that predicted for the stacking of a dangling 3' unpaired adenosine on a G-C or G x U base pair. Group 1A monovalent ions, NH4+, Mg2+, and Co(NH3)6(3+) ions stabilize the A14 and deltaA14 pseudoknots to largely identical extents, revealing that the observed differences in stability in these molecules do not derive from a differential or specific accumulation of ions in the A14 versus deltaA14 pseudoknots. Knowledge of this free energy contribution may facilitate the prediction of RNA pseudoknot formation from primary nucleotide sequence (Gultyaev et al., 1999, RNA 5:609-617).


Emerging norms for the control of emerging epidemics.


title: Asthma and Respiratory Foundation NZ Adolescent and Adult Asthma Guidelines 2020: a quick reference guide.

text: The purpose of the 2020 Asthma and Respiratory Foundation NZ Adolescent and Adult Asthma Guidelines is to provide simple, practical and evidence-based recommendations for the diagnosis, assessment and management of asthma in adolescents and adults (aged 12 and over) in a quick reference format. The intended users are health professionals responsible for delivering asthma care in the community and hospital settings, and those responsible for the training of such health professionals. The main changes in the 2020 update are: 1) combining the recommendations for both adolescents and adults in a single document, 2) the recommendation to avoid SABA-only treatment in the long-term management of asthma, 3) the use of budesonide/formoterol reliever, with or without maintenance budesonide/formoterol, is preferred to SABA reliever, with or without maintenance ICS or ICS/LABA, across the spectrum of asthma severity, 4) introduction of the terminology 'anti-inflammatory reliever (AIR)' therapy to describe the use of budesonide/formoterol as a reliever medication, with or without maintenance budesonide/ formoterol therapy. This approach encompasses and extends the 'Single combination ICS/LABA inhaler Maintenance And Reliever Therapy' (SMART) approach recommended in the previous guideline, 5) the inclusion of two stepwise management algorithms, 6) a clinical allergy section, 7) the role of LAMA therapy in severe asthma, 8) the role of omalizumab in severe allergic asthma and mepolizumab in severe eosinophilic asthma, 9) an appendix detailing educational materials.


title: [Seroepidemiological study of coronavirus infection in children and adults in St. Petersburg].

text: As the result of prolonged (17 years) observations of patients with acute respiratory infections hospitalized in basic departments of clinics of the Research Institute of Influenza, coronavirus infection was found to be the cause of respiratory diseases, on the average, in 12% of cases (in some years in 6.8% to 28.6% of cases). The analysis of extensive morbidity rates among different age groups of the population showed that children were affected by coronavirus infection 5-7 times more often than adults. Three year cycles of this infection were established. The periods of coronaviruses activation were accompanied by their detection in patient material by electron-microscopy, a sharp increase of immune response of patients as well as in the number of nosocomial infections and the proportion of the monoinfection of the coronavirus nature. Coronaviruses played the leading role among other viruses in the etiology of hospital respiratory infections. Mucosal antibodies to coronaviruses in the secretions of the nasal cavity proved to be more important than serum antibodies not only in protection from infection, but also in the pattern of clinical manifestations of the disease.


Background paper. Aspects of coronavirus evolution.


title: The relationship between job stress, burnout and clinical depression.

text: The definition and phenomenological features of 'burnout' and its eventual relationship with depression and other clinical conditions are reviewed. Work is an indispensable way to make a decent and meaningful way of living, but can also be a source of stress for a variety of reasons. Feelings of inadequate control over one's work, frustrated hopes and expectations and the feeling of losing of life's meaning, seem to be independent causes of burnout, a term that describes a condition of professional exhaustion. It is not synonymous with 'job stress', 'fatigue', 'alienation' or 'depression'. Burnout is more common than generally believed and may affect every aspect of the individual's functioning, have a deleterious effect on interpersonal and family relationships and lead to a negative attitude towards life in general. Empirical research suggests that burnout and depression are separate entities, although they may share several 'qualitative' characteristics, especially in the more severe forms of burnout, and in vulnerable individuals, low levels of satisfaction derived from their everyday work. These final issues need further clarification and should be the focus of future clinical research.


title: [Microbiological development studies in a clinically healthy, closed primate colony (Macaca mulatta, Macaca arctoides) after the establishment of a Salmonella infection].

text: A case of a Salmonella infantis septicemia in a primate (Macaca mulatta) probably induced by shipment stress gave rise to microbiological examination of fecal samples from all animals of a since 1976 closed colony of Macaca mulatta and Macaca arctoides. During 3 investigations carried out in 3 week intervals salmonella-positive animals were separated immediately from the stock. At the first examination 3 of 52, and at the second 2 of 49 samples were found to be salmonella-positive. At the third series all of the remaining 47 animals proved to be salmonella-negative. All isolates were identified as S. infantis. Feeding of contaminated, dried prawns was suspected as the most probable source of infection. A simultaneous examination of 96 of the stool specimens revealed Y. enterocolitica in 4 samples. Additionally, electron-microscopy of pooled fecal samples (3 to 4 animals each) led to demonstration of corona-, adenovirus and small round virus-like particles in one pool each. Moreover, coronavirus-like particles could be detected in 7 of 13 (1st examination series), 5 of 15 (2nd series) and 1 of 13 pools (3rd series).


Hong Kong hampered in fight against SARS by lack of cooperation from mainland.


title: Spontaneous Reye's-like syndrome in BALB/cByJ mice.

text: In five spontaneous outbreaks, sixty-four BALB/cByJ mice developed Reye's-like syndrome 4 to 33 days after introduction into mouse rooms known to harbor a variety of indigenous murine viruses. The clinical course lasted 24 hours and consisted of progressively deteriorating consciousness and hyperventilation, usually leading to death. Mice killed and necropsied while stuporous or comatose had serum ammonia levels of 2524 +/- 179 versus 66 +/- 8 micrograms/dl for control BALB/cByJ mice. Necropsy findings included swollen, diffusely pale yellow livers with panlobular microvesicular fatty change, pale renal cortices with epithelial fat droplets of the proximal convoluted tubules, and Alzheimer type II astrocytosis of the neocortex, corpus striatum, hippocampus, and thalamus. In three of the five outbreaks involving 66% of affected animals, mice had active coronaviral enteritis. Electron microscopic changes in hepatocytes, neocortical astrocytes, and neurons were similar to or identical with those described for the acute phase of Reye's syndrome in man. The epizootiology of these outbreaks and the apparent synchrony between Reye's syndrome-like illness and the stage of intestinal coronavirus infections in some of these outbreaks suggest that one or more naturally occurring murine viruses were of etiologic significance.


title: Techniques and cost effectiveness of enteroatmospheric fistula isolation .

text: UNLABELLED  Background. The following describes successful isolation of enteroatmospheric fistulae within a negative pressure wound therapy system (V.A.C.®, KCI, San Antonio, TX). METHODS An impermeable tubular structure was placed around the fistula with a dressing applied to the surrounding wound base, dressed with an impermeable drape and negative pressure, and then an ostomy appliance was placed over the isolated fistula stoma. Cost analysis compared traditional dressings to the NPWT isolation method. RESULTS All patients underwent fistula isolation with complete diversion of enteric contents. Typical dressing changes occurred 3 times per week. Four patients were discharged from the hospital prior to surgical repair. CONCLUSION Successful isolation of enteroatmospheric fistulae using a NPWT system has the potential to lower healthcare system costs by allowing for earlier hospital discharge.


title: Daily Emergency Department Surveillance System --- Bergen County, New Jersey.

text: The purpose of the Daily Emergency Department Surveillance System (DEDSS) is to provide consistent, timely, and robust data that can be used to guide public health activities in Bergen County, New Jersey. DEDSS collects data on all emergency department visits in four hospitals in Bergen County and analyzes them for aberrant patterns of disease or single instances of certain diseases or syndromes. The system monitors for clusters of patients with syndromes consistent with the prodrome of a terrorism-related illness (e.g., anthrax or smallpox) or naturally occurring disease (e.g., pandemic influenza or food and waterborne outbreaks). The health department can use these data to track and characterize the temporal and geographic spread of a known outbreak or demonstrate the absence of cases during the same period (e.g., severe acute respiratory syndrome [SARS] or anthrax). DEDSS was designed to be flexible and readily adaptable as local, state, or federal surveillance needs evolve.


title: Transmission and prevention of occupational infections in orthopaedic surgeons.

text: Microorganisms are transmitted in hospitals mainly by contact, droplet, and airborne routes. Orthopaedic surgeons have a substantial occupational risk of contracting a blood-borne infection because of frequent handling of sharp instruments and objects during operative procedures. Aerosolization means the formation of aerosols and droplets when blood or other body fluids are mechanically disturbed. Smaller particles (<5 microm) will remain suspended in air. Pathogens that can survive in these small airborne particles may cause infection if they are inhaled. Aerosol-generating procedures in patients with tuberculosis or severe acute respiratory syndrome (SARS) may facilitate airborne transmission. The Hospital Infection Control Practices Advisory Committee and the Centers for Disease Control and Prevention have established guidelines for isolation precautions in hospitals.


title: Risk-stratified seroprevalence of SARS coronavirus in children residing in a district with point-source outbreak compared to a low-risk area.

text: 1. SARS coronavirus has low transmissibility at the community level. 2. Subclinical SARS coronavirus infection is rare in children.


title: Point mutations in the S protein connect the sialic acid binding activity with the enteropathogenicity of transmissible gastroenteritis coronavirus.

text: Enteropathogenic transmissible gastroenteritis virus (TGEV), a porcine coronavirus, is able to agglutinate erythrocytes because of sialic acid binding activity. Competitive inhibitors that may mask the sialic acid binding activity can be inactivated by sialidase treatment of virions. Here, we show that TGEV virions with efficient hemagglutinating activity were also obtained when cells were treated with sialidase prior to infection. This method was used to analyze TGEV mutants for hemagglutinating activity. Recently, mutants with strongly reduced enteropathogenicity that have point mutations or a deletion of four amino acids within residues 145 to 155 of the S protein have been described. Here, we show that in addition to their reduced pathogenicity, these mutants also have lost hemagglutinating activity. These results connect sialic acid binding activity with the enteropathogenicity of TGEV.


title: Molecular Epidemiology of Human Metapneumovirus in Riyadh Province, Saudi Arabia.

text: Human metapneumovirus (HMPV) is an important cause of respiratory tract illness in children. Two HMPV subgroups, A and B, and four genotypes, A1, A2, B1 and B2, have been identified. Concurrent circulation of the different genotypes in yearly epidemics has been recorded globally, but not in Saudi Arabia. The current report was designed to study HMPV epidemiology in Saudi children and to analyze the genetic diversity and circulation patterns. Nasopharyngeal aspirates (n = 174) were collected from hospitalized children in Riyadh (2008-2009). The screening of samples using real-time RT-PCR identified 19 HMPV strains. The majority of the strains belonged to subgroup B, while all strains of subgroup A were members of genotype A2. In 2008, only subgroup B was recognized, whereas in 2009 both subgroups were identified to be cocirculating at similar rates. The full-length attachment (G) gene and a partial sequence of the fusion (F) gene of positive samples were sequenced. The G gene showed a high degree of genetic diversity and exhibited a variable number of positively selected sites in different lineages. In contrast, the F gene demonstrated an extensive genetic stability with a higher tendency toward purifying selection. This is the first report on HMPV genotype circulation in Saudi Arabia; however, the exact circulation kinetics requires further retrospective and prospective study.


title: Infection control for the otolaryngologist in the era of severe acute respiratory syndrome.

text: OBJECTIVES Severe acute respiratory syndrome (SARS) has affected more than 8400 persons in 28 countries, with more than 800 deaths. The current SARS outbreak, especially in North American health care centers, has motivated a re-evaluation of infection control practices in the hospital and clinic environment. These considerations are particularly important to otolaryngology, in which examination and diagnostic procedures often bring the otolaryngologist in close--if not direct--contact with the patient's upper airway, mucosa, and secretions. The otolaryngologist is at increased risk of contracting a respiratory pathogen. METHOD A joint effort by the Department of Otolaryngology at Queen's University and the Infection Control Services of the Hotel Dieu Hospital, Kingston, Ontario, was carried out to develop specific infection control guidelines for the otolaryngologists using strategies from the Centers for Disease Control and Prevention in the United States and the Laboratory Center for Disease Control, Health Canada. RESULTS A set of specific recommendations was developed for the otolaryngologists to augment current infection control, including diligent use of personal protective equipment with every patient encounter. Moreover, this equipment should be removed according to specific protocol, to avoid contamination of self, others, and surroundings. Finally, a number of practice modifications are being adopted as prudent precautionary measures. CONCLUSION It is essential to adhere to these recommendations in order to protect the health and safety of clinicians, colleagues, and patients.


title: Public Health Laboratory System Improvement Program: development and implementation.

text: Instruments designed to measure the performance of public health systems at state and local levels were supported by the Centers for Disease Control and Prevention (CDC) and implemented in 2002. This article describes the process and outcomes of a system and tool designed to measure performance of State Public Health Laboratory Systems, accomplished by the Association of Public Health Laboratories (APHL) in partnership with CDC. We describe the process used to develop the instrument and its subsequent pilot testing and field testing in 11 states. Throughout the field testing and early implementation phases, both CDC and APHL recognized that the core rationale for measuring system performance would be to provide the basis for subsequent system improvement. APHL implemented the Laboratory System Improvement Program (L-SIP) in 2007 and conducted an evaluation of the field testing of the instrument and related materials that same year. We conclude with a summary of future implications for L-SIP, the program's recognition as an international standard for laboratory systems, and the critical importance of its continuation.


title: Seasonal influenza and 2009 H1N1 influenza vaccination coverage among pregnant women--10 states, 2009-10 influenza season.

text: Because pregnant women are at increased risk for severe disease associated with influenza infection, the American College of Obstetricians and Gynecologists and the Advisory Committee on Immunization Practices have recommended seasonal influenza vaccination for women while pregnant, regardless of trimester. In 2009, a novel strain of influenza A (H1N1) virus was identified, and pregnant women also were found to be at greater risk for influenza-related complications from this new virus. As a result, during the 2009--10 influenza season, two separate influenza vaccines were recommended to pregnant women: inactivated trivalent 2009--10 seasonal vaccine and influenza A (H1N1) 2009 monovalent vaccine. To estimate influenza vaccination coverage among pregnant women during the 2009--10 influenza season, CDC analyzed data from 10 states from the Pregnancy Risk Assessment Monitoring System (PRAMS). This report summarizes the results of that analysis, which determined that vaccination coverage for pregnant women among the 10 states combined was 50.7% for seasonal influenza and 46.6% for 2009 H1N1. In addition, women to whom vaccination was offered or recommended by their health-care provider were significantly more likely to report being vaccinated against seasonal influenza (relative risk [RR] = 3.3) and 2009 H1N1 (RR = 10.1). These results indicate substantially higher influenza vaccination coverage among pregnant women than has been reported for previous influenza seasons and support previous findings that receipt of influenza vaccination can be influenced greatly by health-care providers offering or recommending influenza vaccination.


title: Anti-SARS coronavirus 3C-like protease effects of Rheum palmatum L. extracts.

text: The present study aims to clarify the inhibitive effect of the compounds from Rheum palmatum L. on the SARS-3CL protease. The SARS-CoV 3CL gene was amplified from RNA of the SARS virus by PCR. The SARS-CoV 3CL protease was purified from a colon bacillus recombinant. Drugs and 3CL protease were incubated together. The inhibition rate and IC(50) were calculated based on absorbance. Components from the Rheum palmatum L. had a high level of anti-SARS-CoV 3CL protease activity. The IC(50) was 13.76 +/- 0.03 microg/mL and the inhibition rate was up to 96%. In conclusion, extracts from Rheum palmatum L. have a high level of inhibitory activity against 3CL protease, suggesting that extracts from Rheum palmatum L. may represent a potential therapeutic for SARS.


Feline infectious uveitis.


title: [Conditions for SARS-CoV cultivation and inactivation].

text: OBJECTIVE To study the method for cultivation and inactivation of SARS-CoV. METHODS In order to choose the sensitive cell strain and the best infection dose of the virus, Vero, Vero-E6 and 2BS cell lines were infected with SARS-CoV. The cultivation temperature was selected among 25 degrees C, 33 degrees C and 37 degrees C. The best inactivating time and effect were observed with beta-propiolactone whose concentration ranged from 1:2000 to 1:20,000 at room temperature. RESULTS Vero and Vero-E6 cell lines were sensitive to SARS-CoV. The cytopathic changes of the cells were 75% at 37 degrees C in 5 percent CO2 incubator after infection. SARS-CoV was inactivated completely in beta-propiolactone (1:4000). The toxicity of beta-propiolactone was hydrolyzed completely when the inactivated virus was cultured for 16 hours at 2 degrees C, 8 degrees C and in water bath for 2 hours at 37 degrees C. CONCLUSION The titer of SARS-CoV was the highest when it was cultured in Vero or Vero-E6 cells for 72 hours at 37 degrees C in 5 percent CO2 incubator. SARS-CoV was inactivated completely in beta-propiolactone when its concentration was 1:4000 and the interaction time was 1 hour.


title: Transmissible gastroenteritis coronavirus packaging signal is located at the 5' end of the virus genome.

text: To locate the transmissible gastroenteritis coronavirus (TGEV) packaging signal, the incorporation of TGEV subgenomic mRNAs (sgmRNAs) into virions was first addressed. TGEV virions were purified by three different techniques, including an immunopurification using an M protein-specific monoclonal antibody. Detection of sgmRNAs in virions by specific reverse transcription-PCRs (RT-PCRs) was related to the purity of virus preparations. Interestingly, virus mRNAs were detected in partially purified virus but not in virus immunopurified using stringent conditions. Analyses by quantitative RT-PCR confirmed that virus mRNAs were not present in highly purified preparations. Lack of sgmRNA encapsidation was probably due to the absence of a packaging signal (Psi) within these mRNAs. This information plus that from the encapsidation of a collection of TGEV-derived minigenomes suggested that Psi is located at the 5' end of the genome. To confirm that this was the case, a set of minigenomes was expressed that included an expression cassette for an mRNA including the beta-glucuronidase gene (GUS) plus variable sequence fragments from the 5' end of the virus genome potentially including Psi. Insertion of the first 649 nucleotides (nt) of the TGEV genome led to the specific encapsidation of the mRNA, indicating that a Psi was located within this region which was absent from all of the other virus mRNAs. The presence of this packaging signal was further confirmed by showing the expression and rescue of the mRNA including the first 649 nt of the TGEV genome under control of the cytomegalovirus promoter in TGEV-infected cells. This mRNA was successfully amplified and encapsidated, indicating that the first 649 nt of TGEV genome also contained the 5' cis-acting replication signals. The encapsidation efficiency of this mRNA was about 30-fold higher than the genome encapsidation efficiency, as estimated by quantitative RT-PCR. In contrast, viral mRNAs presented significantly lower encapsidation efficiencies (about 100-fold) than those of the virus genome, strongly suggesting that TGEV mRNAs in fact lacked an alternative TGEV Psi.


title: Working memory impairment in pilots exposed to acute hypobaric hypoxia.

text: INTRODUCTION During an acute hypoxia exposure, impairment of memory is one of the most frequently reported symptoms, either during hypoxia awareness training of aircrews or after an in-flight hypoxic incident. However, the effects of acute hypoxia on memory have been little studied in laboratory-controlled conditions. Moreover, none of these studies were performed in hypobaric conditions. The main aim of our study was to investigate the effects of acute hypobaric hypoxia on working memory (WM). This study also aimed to find links between physiological measurements and cognitive performance during acute hypoxia exposure. METHODS During hypoxia awareness training, 28 subjects (experimental group) were exposed to a simulated altitude level of 10,000 m (31,000 ft) in a hypobaric chamber, while 29 subjects (control group) stayed at sea level. WM was assessed in both groups with the Paced Auditory Serial Addition Test (PASAT). Peripheral oxygen saturation (SpO2) and heart rate were recorded. RESULTS WM was strongly impaired in the hypoxic group. One major finding is that hypoxia highly increased the mean error frequency rate. WM performance decreased linearly with hypoxemia, but SpO2 was weakly predictive of PASAT performance and vice versa. DISCUSSION WM is impaired by acute hypobaric hypoxia. Given the importance of WM in aircraft piloting and its sensitivity to hypoxia, the PASAT, in association with SpO2 and EEG recordings, could improve both hypoxia training and our understanding of the effects of hypoxia on memory.


title: Comparative evaluation of real-time PCR assays for detection of the human metapneumovirus.

text: The human metapneumovirus (hMPV) is a new member of the Paramyxoviridae family associated with acute respiratory tract infections in humans. The objective of this study was to compare the sensitivity of real-time RT-PCR assays performed in a LightCycler instrument and designed to amplify the viral nucleoprotein (N), matrix (M), fusion (F), phosphoprotein (P), and polymerase (L) genes. In a first evaluation of 20 viral cultures with characteristics compatible with hMPV cytopathic effect, the PCR positivity rates were 100, 90, 75, 60, and 55% using primers for the N, L, M, P, and F genes. In a second evaluation of 10 nasopharyngeal aspirates from children with bronchiolitis and found to be positive for the hMPV N gene, the PCR positivity rates for the L, M, P, and F genes were 90, 60, 30, and 80%, respectively. The analytic sensitivity of the real-time RT-PCR assay for the hMPV N gene was 100 copies using a transcribed viral plasmid. In conclusion, real-time PCR assays aimed at amplifying the N and L genes which are coding for two internal viral proteins appear particularly suitable for hMPV diagnostic.


title: Attitudes to living and working in pandemic conditions among emergency prehospital medical care personnel.

text: INTRODUCTION Little is known about the risk perceptions and attitudes of healthcare personnel, especially of emergency prehospital medical care personnel, regarding the possibility of an outbreak or epidemic event. PROBLEM This study was designed to investigate pre-event knowledge and attitudes of a national sample of the emergency prehospital medical care providers in relation to a potential human influenza pandemic, and to determine predictors of these attitudes. METHODS Surveys were distributed to a random, cross-sectional sample of 20% of the Australian emergency prehospital medical care workforce (n = 2,929), stratified by the nine services operating in Australia, as well as by gender and location. The surveys included: (1) demographic information; (2) knowledge of influenza; and (3) attitudes and perceptions related to working during influenza pandemic conditions. Multiple logistic regression models were constructed to identify predictors of pandemic-related risk perceptions. RESULTS Among the 725 Australian emergency prehospital medical care personnel who responded, 89% were very anxious about working during pandemic conditions, and 85% perceived a high personal risk associated with working in such conditions. In general, respondents demonstrated poor knowledge in relation to avian influenza, influenza generally, and infection transmission methods. Less than 5% of respondents perceived that they had adequate education/training about avian influenza. Logistic regression analyses indicate that, in managing the attitudes and risk perceptions of emergency prehospital medical care staff, particular attention should be directed toward the paid, male workforce (as opposed to volunteers), and on personnel whose relationship partners do not work in the health industry. CONCLUSIONS These results highlight the potentially crucial role of education and training in pandemic preparedness. Organizations that provide emergency prehospital medical care must address this apparent lack of knowledge regarding infection transmission, and procedures for protection and decontamination. Careful management of the perceptions of emergency prehospital medical care personnel during a pandemic is likely to be critical in achieving an effective response to a widespread outbreak of infectious disease.


Labs form a new front against deadly pathogens.


title: Characterization of the stunting syndrome agent: relatedness to known viruses.

text: An enteric disease of young turkeys, referred to as stunting syndrome (SS), causes reduced growth and impaired feed efficiency. A recently isolated virus, stunting syndrome agent, (SSA) has been found to be the etiologic agent of SS. The objective of the present study was to determine relatedness of the SSA with other viral agents. Serologic (viral neutralization and enzyme-linked immunosorbent assay [ELISA]) assays and a reverse transcriptase-polymerase chain reaction (RT-PCR) were used. The antisera against turkey enteric coronavirus (bluecomb agent), bovine coronavirus (BCV), bovine Breda-1 virus, bovine Breda-2 virus, avian infectious bronchitis virus (IBV), avian influenza virus, Newcastle disease virus (NDV), and transmissible gastroenteritis virus (TGEV) of swine were evaluated by dot-immunobinding avidin-biotin-enhanced ELISA and did not react with SSA. The homologous (anti-SSA) antiserum was positive by ELISA. Similarly, anti-SSA antiserum did not react when NDV, IBV, BCV, or TGEV was used as antigen but did react with the homologous (SSA) virus. The virus neutralization assay was performed by inoculating 24-to-25-day-old turkey embryos via the amniotic route and by assessing the embryo infectivity on the basis of gross intestinal lesions and intestinal maltase activity at 72 hr postinoculation. None of the aforementioned antisera neutralized SSA infectivity in embryos except for the homologous anti-SSA antiserum. A RT-PCR was performed with known primers specific for NDV, IBV, BCV, and TGEV. The known primers failed to amplify SSA genome but amplified their respective viral genomes. We concluded that the SSA was distinct from the viral agents that were evaluated.


title: Trends in childhood cancer mortality--United States, 1990-2004.

text: Cancer is the fourth most common cause of death (after unintentional injury, homicide, and suicide) among persons aged 1-19 years in the United States. Because recent childhood cancer mortality has not been well characterized in terms of temporal, demographic, and geographic trends, CDC analyzed cancer death rates among children (defined as aged 0-14 years) and adolescents (defined as aged 15-19 years) for the period 1990-2004 by sex, age group, race, ethnicity, U.S. Census region, and primary cancer site/leading diagnosis, using the most recent data available from the National Vital Statistics System (NVSS). This report describes the results of that analysis, which indicated that, overall, age-adjusted childhood cancer death rates decreased significantly during 1990-2004 among both sexes, both age groups, all races (except American Indians/Alaska Natives [AI/ANs]), Hispanics, non-Hispanics, and all U.S. Census regions. However, decreases in death rates varied among U.S. Census regions and between Hispanics and non-Hispanics. Eliminating racial/ethnic health disparities is one of the overarching goals of Healthy People 2010. Further research is needed to understand geographic and ethnic disparities in childhood cancer death rates. Moreover, cancer prevention and intervention measures should be designed to reach populations that are underserved and at high risk.


title: Neutralization of enteric coronaviruses with Escherichia coli cells expressing single-chain Fv-autotransporter fusions.

text: We report here that fusions of single-chain antibodies (scFvs) to the autotransporter beta domain of the IgA protease of Neisseria gonorrhoeae are instrumental in locating virus-neutralizing activity on the cell surface of Escherichia coli. E. coli cells displaying scFvs against the transmissible gastroenteritis coronavirus on their surface blocked in vivo the access of the infectious agent to cultured epithelial cells. This result raises prospects for antiviral strategies aimed at hindering the entry into target cells by bacteria that naturally colonize the same intestinal niches.


Diarrhoea in foals.


Correction in identification of a coronavirus-like agent isolated from parrots.


title: Strategies to optimize the performance of Robotic-assisted laparoscopic hysterectomy.

text: A hybrid technique of robot-assisted, laparoscopic hysterectomy using the ENSEAL(®) Tissue Sealing Device is described in a retrospective, consecutive, observational case series. Over a 45 month period, 590 robot-assisted total laparoscopic hysterectomies +/- oophorectomy for benign and malignant indications were performed by a single surgeon with a bedside assistant at a tertiary healthcare center. Patient demographics, indications for surgery, comorbidities, primary and secondary surgical procedures, total operative and surgical time, estimated blood loss (EBL), length of stay (LOS), complications, transfusions and subsequent readmissions were analyzed. The overall complication rate was 5.9% with 35 patients experiencing 69 complications. Mean (SD) surgery time, operating room (OR) time, EBL, and LOS for the entire cohort were 75.5 (39.42) minutes, 123.8 (41.15) minutes, 83.1 (71.29) millilitres, and 1.2 (0.93) days, respectively. Mean surgery time in the first year (2009) was 91.6 minutes, which declined significantly each year by 18.0, 19.0, and 24.3 minutes, respectively. EBL and LOS did not vary -significantly across the entire series. Using the cumulative sum method, an optimization curve for surgery time was evaluated, with three distinct optimization phases observed. In summary, the use of an advanced laparoscopic tissue-sealing device by a bedside surgical assistant provided an improved operative efficiency and reliable vessel sealing during robotic hysterectomy.


title: Cell culture propagation of a coronavirus isolated from cows with winter dysentery.

text: Fecal filtrates from cows with winter dysentery were inoculated into gnotobiotic and conventional calves, and a coronavirus was isolated from calf feces. Cytopathic effects were observed on human rectal tumor cells but not bovine cell cultures. The winter dysentery isolates morphologically and antigenically resembled the Mebus strain of bovine coronavirus.


title: [Translational control of the picornavirus phenotype].

text: Picornaviruses are small animal viruses with positive-stranded genomic RNA, which is translated using cap-independent internal translation initiation. The key role in this is played by cis elements of the 5'-untranslated region (5'-UTR) and, in particular, by the internal ribosome entry site (IRES). The function of translational cis elements requires both canonical translation initiation factors (eIFs) and additional IRES trans-acting factors (ITAFs). All known ITAFs are cell RNA-binding proteins which play a variety of functions in noninfected cells. Specific features of translational cis elements substantially affect the phenotype and, in particular, tissue tropism and pathogenic properties of picornaviruses. It is clear that, in some cases, the molecular mechanism of this is a change in interactions between viral cis elements and ITAFs. The properties and tissue distribution of ITAFs may determine the biological properties of other viruses that also use the IRES-dependent translation initiation. Since this mechanism is also involved in translation of several cell mRNAs, ITAF may contribute to the regulation of the most important aspects of the living activity in noninfected cells.


Infection control of pandemic (H1N1) 2009 influenza in hospitals - a logistic challenge.


title: Robotic Pancreaticoduodenectomy: Operative Steps (With Video).

text: Robotic pancreaticoduodenectomy represents actually an emerging procedure for robot-assisted surgery. Considering complexity, extensive dissection, and difficult restoration of the digestive continuity, it remains a challenge for surgeons. The embedded video (Supplemental Digital Content 1, http://links.lww.com/SLE/A146) reports our standardized technique for robotic pancreaticoduodenectomy. In conclusion, robotic pancreaticoduodenectomy is feasible and can be safely performed. Further evaluation with clinical trials is required to validate its real benefits.


Endoscopic Ultrasound-Guided Biliary Drainage for Palliation of Malignant Obstructive Jaundice.


Fibromyalgia after severe acute respiratory syndrome: a case report.


iNOS (NOS2) at a glance.


title: Relationship between preservation of the lateral pillar and collapse of the femoral head in patients with osteonecrosis.

text: It is difficult to predict if and when the femoral head will collapse and whether the collapse can be minimized. This study examined the final outcome of early-stage osteonecrosis of the femoral head (ONFH) using magnetic resonance imaging to verify the relationship between preservation of the lateral pillar and collapse of the femoral head. A midcoronal section of the femoral head was divided into 3 pillars (medial, central, and lateral) on a T1-weighted image. According to the site of necrosis on the lateral pillar, the necrosis was divided into 3 types: I, the necrosis occupies the central and medial pillars, and the lateral pillar is preserved; II, the necrosis partially occupies the lateral pillar; and III, the necrosis totally occupies the lateral pillar. One group of 87 patients (127 hips) with Association for Research on Osseous Circulation (ARCO) stage I ONFH underwent nonoperative treatment and were followed for 3 to 8 years (average, 6.2 years). Another group of 42 patients (72 hips) with ARCO stage I ONFH underwent debridement and impacted bone graft and were followed for 5 to 9 years (average, 7.1 years). In both groups, the more preserved the lateral pillar, the less collapse occurred. The authors concluded that whether ONFH progressed to collapse is determined by preservation of the lateral pillar. The lateral pillar is the keystone for maintaining the sphere of the femoral head and its preservation.


title: Literature review: the common cold.

text: Most colds are caused by rhinovirus infection, perhaps facilitated by chilling or stress. Virus infection begins in the nasopharynx and causes spotty destruction of the nasal ciliated epithelium. Transmission occurs chiefly via droplets of various sizes transported through the air, but some types of virus persist in moist secretions on handled objects and may retain their infectiousness. Living in crowded, poorly-ventilated quarters facilitates transmission. Not many virus particles survive in saliva and it is difficult to infect via the lips or mouth. Kissing does not efficiently spread cold infection. Prophylactic treatment with interferon does not protect against cold infection. Aspirin and acetaminophen reduced serum antibody response and increased nasal symptoms in a controlled Australian study. The combination of intranasal interferon and ipratropium with oral naproxen gave promising results in experimental rhinovirus inoculation. Basically, there has been little or no progress towards effective cold treatment in the past century.


title: Cross-host evolution of severe acute respiratory syndrome coronavirus in palm civet and human.

text: The genomic sequences of severe acute respiratory syndrome coronaviruses from human and palm civet of the 2003/2004 outbreak in the city of Guangzhou, China, were nearly identical. Phylogenetic analysis suggested an independent viral invasion from animal to human in this new episode. Combining all existing data but excluding singletons, we identified 202 single-nucleotide variations. Among them, 17 are polymorphic in palm civets only. The ratio of nonsynonymous/synonymous nucleotide substitution in palm civets collected 1 yr apart from different geographic locations is very high, suggesting a rapid evolving process of viral proteins in civet as well, much like their adaptation in the human host in the early 2002-2003 epidemic. Major genetic variations in some critical genes, particularly the Spike gene, seemed essential for the transition from animal-to-human transmission to human-to-human transmission, which eventually caused the first severe acute respiratory syndrome outbreak of 2002/2003.


title: Virus complement evasion strategies.

text: The immune system has a variety of tools at its disposal to combat virus infections. These can be subdivided roughly into two categories: 'first line defence', consisting of the non-specific, innate immune system, and 'adaptive immune response', acquired over time following virus infection or vaccination. During evolution, viruses have developed numerous, and often very ingenious, strategies to counteract efficient recognition of virions or virus-infected cells by both innate and adaptive immunity. This review will focus on the different strategies that viruses use to avoid recognition by one of the components of the immune system: the complement system. Complement evasion is of particular importance for viruses, since complement activation is a crucial component of innate immunity (alternative and mannan-binding lectin activation pathway) as well as of adaptive immunity (classical, antibody-dependent complement activation).


title: Nucleotide sequence shows that Bean leafroll virus has a Luteovirus-like genome organization.

text: The complete nucleotide sequence of the Bean leafroll virus (BLRV) genomic RNA and the termini of its smallest subgenomic RNAs were determined to better understand its mechanisms of gene expression and replication and its phylogenetic position within the Luteoviridae: The number and placement of open reading frames (ORFs) within the BLRV genome was Luteovirus-like. The nucleotide and predicted amino acid sequences of BLRV were most similar to those of Soybean dwarf virus (SbDV). Phylogenetic analyses employing the neighbour-joining method and sister-scanning analysis indicated that the BLRV nonstructural proteins were closely related to those of Barley yellow dwarf virus-PAV (BYDV-PAV), a luteovirus: The region surrounding the frameshift at the junction between ORFs 1 and 2 also contained sequences very similar to those of BYDV-PAV and a Dianthovirus, Red clover necrotic mosaic virus. Similar analyses showed that the structural proteins were most similar to those of the Polerovirus genus. The 3'-noncoding regions downstream of ORF5 contained sequences similar to translational control elements identified in the BYDV-PAV genome. These data suggest that BLRV, like SbDV, is derived either through selection from a common ancestor with BYDV-PAV or that BLRV is the product of two recombination events between luteovirus-like and polerovirus-like ancestors where the 5' 2900 nt and 3' 700 nt of the BLRV genome are from a Luteovirus and the intervening sequences are derived from a Polerovirus:


title: [Asthma and COPD management during the COVID-19 pandemic].

text: Numerous patients with asthma or COPD are likely to be infected with SARS-CoV-2 virus. Although data is limited, patients with severe and/or uncontrolled asthma and those with COPD appear to be at increased risk of a more severe course of COVID-19 infection. Usual recommendations for management of asthma and COPD remain valid despite the ongoing epidemic. However, lung function testing and nebulisers should be performed with caution during the COVID-19 pandemic due to a potential risk of virus aerosolisation and contagion during the procedure. Particular care must be taken to identify and protect patients who are particularly vulnerable to COVID-19 infection. Asthma and COPD treatments should be pursued and adapted to ensure optimal control of the lung disease throughout the epidemic, thus reducing the risk of severe COVID-19 disease.


title: Report of the European Society of Cardiology Cardiovascular Round Table regulatory workshop update of the evaluation of new agents for the treatment of acute coronary syndrome: Executive summary.

text: Regulatory authorities interpret the results of randomized controlled trials according to published principles. The European Medicines Agency (EMA) is planning a revision of the 2000 and 2003 guidance documents on clinical investigation of new medicinal products for the treatment of acute coronary syndrome (ACS) to achieve consistency with current knowledge in the field. This manuscript summarizes the key output from a collaborative workshop, organized by the Cardiovascular Round Table and the European Affairs Committee of the European Society of Cardiology, involving clinicians, academic researchers, trialists, European and US regulators, and pharmaceutical industry researchers. Specific questions in four key areas were selected as priorities for changes in regulatory guidance: patient selection, endpoints, methodologic issues and issues related to the research for novel agents. Patients with ST-segment elevation myocardial infarction (STEMI) and non-STEMI (NSTEMI) should be studied separately for therapies aimed at the specific pathophysiology of either condition, particularly for treatment of the acute phase, but can be studied together for other treatments, especially long-term therapy. Unstable angina patients should be excluded from acute phase ACS trials. In general, cardiovascular death and reinfarction are recommended for primary efficacy endpoints; other endpoints may be considered if specifically relevant for the therapy under study. New agents or interventions should be tested against a background of evidence-based therapy with expanded follow-up for safety assessment. In conclusion, new guidance documents for randomized controlled trials in ACS should consider changes regarding patient and endpoint selection and definitions, and trial designs. Specific requirements for the evaluation of novel pharmacological therapies need further clarification.


title: Airway inflammation and upper respiratory tract infection in athletes: is there a link?

text: Upper Respiratory Tract Infection (URTI) is regarded as the most common medical condition affecting both highly trained and elite athletes, in particular those participating in endurance events. The causes of these disturbances, also occurring during training, remain unclear. Viruses such as rhinovirus, adenovirus and para-influenza virus are frequently reported as the source of URTI. However, in a few comprehensive laboratory and epidemiological studies which reported at least a 30% incidence of URTI, no identifiable pathogens were either reported or studied. A recent, longitudinal study investigated symptomatology and pathogenic etiology in sedentary controls, recreational and elite athletes. The highest incidence of URTI occurred in elite athletes. However; only 11 out of 37 illness episodes overall had pathogenic origins, and most of the unidentified upper respiratory illnesses were shorter in duration and less severe than infectious ones. This concept of inflammation without infection in athletes is quite new and leads us to consider other explanatory pathophysiological conditions. Increases in airway neutrophils, eosinophils and lymphocytes have been described under resting conditions in endurance sports, swimmers and cross-country skiers. These inflammatory patterns may be due to pollutants or chlorine-related compounds in swimmers. After intense exercise similar airways cellular profiles have been reported, with a high amount of bronchial epithelial cells. This increase in airway inflammatory cells in athletes can result from a hyperventilation-induced increase in airway osmolarity stimulating bronchial epithelial cells to release chemotactic factors. Fortunately, in most cases, these inflammatory cells express rather low level of adhesion molecules, explaining why airway inflammation may appear blunted in athletes despite numerous inflammatory cellular elements. However it can be hypothesized that a transient loss of control of this local inflammation, due to various external physico-chemical strains, might occur. This might account for some of the unidentified upper respiratory illnesses.


title: Mutations in SARS-CoV-2 viral RNA identified in Eastern India: Possible implications for the ongoing outbreak in India and impact on viral structure and host susceptibility.

text: Direct massively parallel sequencing of SARS-CoV-2 genome was undertaken from nasopharyngeal and oropharyngeal swab samples of infected individuals in Eastern India. Seven of the isolates belonged to the A2a clade, while one belonged to the B4 clade. Specific mutations, characteristic of the A2a clade, were also detected, which included the P323L in RNA-dependent RNA polymerase and D614G in the Spike glycoprotein. Further, our data revealed emergence of novel subclones harbouring nonsynonymous mutations, viz. G1124V in Spike (S) protein, R203K, and G204R in the nucleocapsid (N) protein. The N protein mutations reside in the SR-rich region involved in viral capsid formation and the S protein mutation is in the S2 domain, which is involved in triggering viral fusion with the host cell membrane. Interesting correlation was observed between these mutations and travel or contact history of COVID-19 positive cases. Consequent alterations of miRNA binding and structure were also predicted for these mutations. More importantly, the possible implications of mutation D614G (in SD domain) and G1124V (in S2 subunit) on the structural stability of S protein have also been discussed. Results report for the first time a bird's eye view on the accumulation of mutations in SARS-CoV-2 genome in Eastern India.


title: Laboratory safety aspects of SARS at Biosafety Level 2.

text: The severe acute respiratory syndrome (SARS)-associated coronavirus causes severe disease, is transmissible to the community and there is no effective prophylaxis or treatment--perhaps fulfilling the criteria for biohazard group 3 or 4. The recommendation to use Biosafety Level (BSL)3 practices within a BSL2 environment appears to have been a practical decision based on available resources; most diagnostic laboratories operate at BSL2. Safety is achieved with controls in administration, engineering and personal protective equipment/behaviour. At the heart of every safety policy is a risk assessment based on the exact manipulations employed. Excessive administrative and engineering controls are less important than the training and personal attitudes, abilities and understanding of the staff. The SARS outbreak focused our attention on the safety aspects of common mundane tasks, such as decapping blood tubes. Laboratories often claim they follow certain practices but casual observation does not always support these claims. Guidelines differed and created uncertainty. This was stressful for laboratory staff held accountable for their implementation. Attempts to categorise risks and their management into neatly wrapped parcels are attractive, but closer inspection reveals a subjective element that allows doubt to creep in with varying interpretations of the literature. Staff most at risk were those handling respiratory samples. Staff receiving samples via pneumatic tubes had least control over their exposure and were potentially exposed to aerosols from leaking samples. Risk assessment remains a balance between cost and benefit.


title: Disrupting the transmission of influenza a: face masks and ultraviolet light as control measures.

text: In the event of an influenza pandemic, where effective vaccine and antiviral drugs may be lacking, disrupting environmental transmission of the influenza virus will be the only viable strategy to protect the public. We discuss 2 such modalities, respirators (face masks) and ultraviolet (UV) light. Largely overlooked, the potential utility of each is underappreciated. The effectiveness of disposable face masks may be increased by sealing the edges of the mask to the face. Reusable masks should be stockpiled, because the supply of disposable masks will likely prove inadequate. UV light, directed overhead, may be beneficial in hospitals and nursing homes.


title: Neonatal rhinitis.

text: Neonatal rhinitis as a distinct disease entity has not been well-studied. The recognition and treatment of this condition is important since neonates are obligate nasal breathers, and mismanagement of this entity can result in poor feeding or even death from respiratory distress. We undertook a retrospective analysis of 20 patients seen at the Children's Hospital of San Diego over the period 1990-1991. Eighteen patients developed neonatal rhinitis in the months of August to January and only two between February and July. Clinical presentation and an effective management algorithm are discussed. Early recognition on the basis of clinical features followed by a two-step therapeutic trial consisting of conservative therapy and corticosteroid drops are advocated. Based on the above findings we have defined neonatal rhinitis as mucoid rhinorrhea with nasal mucosal edema in the afebrile newborn that results in stertor, poor feeding and respiratory distress which responds promptly to decadron 0.1% drops within a week. We recommend reserving diagnostic procedures for complicated cases that do not respond to the proposed regimen.


title: Outcomes and prognostic factors in 267 patients with severe acute respiratory syndrome in Hong Kong.

text: BACKGROUND Severe acute respiratory syndrome (SARS) has become a global public health emergency. OBJECTIVE To evaluate the characteristics and outcomes of patients with SARS in Hong Kong and to identify predictors of mortality. DESIGN Retrospective cohort study. SETTING Quarantine hospital for patients with SARS in Hong Kong. PATIENTS 267 consecutive patients hospitalized from 26 February to 31 March 2003 for probable or confirmed SARS. MEASUREMENTS Clinical, laboratory, and radiographic measures; 3-month mortality rate. RESULTS According to our case definition, there were 227 cases of confirmed SARS and 40 cases of probable SARS. Common presenting symptoms were fever (99% of patients), chills (74%), malaise (63%), and myalgia (50%). Laboratory findings included lymphopenia (73%), thrombocytopenia (50%), hyponatremia (60%), and elevated levels of lactate dehydrogenase (47%) and C-reactive protein (75%). During hospitalization, incidence of diarrhea (53%), anemia (53%), and acute renal failure (6%) increased. Sixty-nine patients (26%) required intensive care because of respiratory failure. The 3-month mortality rate was 12% (95% CI, 8% to 16%). Factors contributing to mortality were respiratory failure, acute renal failure, and nosocomial sepsis. On multivariate Cox regression, age older than 60 years (relative risk, 5.10 [CI, 2.30 to 11.31]; P < 0.001) and lactate dehydrogenase level greater than 3.8 micro kat/L at presentation (relative risk, 2.20 [CI, 1.03 to 4.71]; P = 0.04) were independent predictors of mortality. CONCLUSION Because of the longer follow-up period in our cohort, the mortality rate in these patients is higher than rates reported in previous studies. Advanced age and high lactate dehydrogenase level at presentation predict mortality. *For members of the Princess Margaret Hospital SARS Study Group, see the Appendix.


title: Using a resequencing microarray as a multiple respiratory pathogen detection assay.

text: Simultaneous testing for detection of infectious pathogens that cause similar symptoms (e.g., acute respiratory infections) is invaluable for patient treatment, outbreak prevention, and efficient use of antibiotic and antiviral agents. In addition, such testing may provide information regarding possible coinfections or induced secondary infections, such as virally induced bacterial infections. Furthermore, in many cases, detection of a pathogen requires more than genus/species-level resolution, since harmful agents (e.g., avian influenza virus) are grouped with other, relatively benign common agents, and for every pathogen, finer resolution is useful to allow tracking of the location and nature of mutations leading to strain variations. In this study, a previously developed resequencing microarray that has been demonstrated to have these capabilities was further developed to provide individual detection sensitivity ranging from 10(1) to 10(3) genomic copies for more than 26 respiratory pathogens while still retaining the ability to detect and differentiate between close genetic neighbors. In addition, the study demonstrated that this system allows unambiguous and reproducible sequence-based strain identification of the mixed pathogens. Successful proof-of-concept experiments using clinical specimens show that this approach is potentially very useful for both diagnostics and epidemic surveillance.


title: Reverse genetics of coronaviruses using vaccinia virus vectors.

text: In this article, we describe the reverse genetic system that is based on the use of vaccinia virus cloning vectors. This system represents a generic approach to coronavirus reverse genetics and was first described for the generation of recombinant human coronavirus 229E representing a group I coronavirus. Subsequently, the same approach has been used to generate recombinant avian infectious bronchitis coronavirus and, recently, recombinant mouse hepatitis virus, representing group III and group II coronaviruses, respectively. We describe how vaccinia virus-mediated homologous recombination can be used to introduce specific mutations into the coronavirus genomic cDNA during its propagation in vaccinia virus and how recombinant coronaviruses can be isolated. Finally, we describe how the coronavirus reverse genetic system has now been extended to the generation of coronavirus replicon RNAs.


title: Glycosaminoglycans and infection.

text: Glycosaminoglycans (GAGs) are complex linear polysaccharides expressed in intracellular compartments, at the cell surface, and in the extracellular environment where they interact with various molecules to regulate many cellular processes implicated in health and disease. Subversion of GAGs is a pathogenic strategy shared by a wide variety of microbial pathogens, including viruses, bacteria, parasites, and fungi. Pathogens use GAGs at virtually every major portals of entry to promote their attachment and invasion of host cells, movement from one cell to another, and to protect themselves from immune attack. Pathogens co-opt fundamental activities of GAGs to accomplish these tasks. This ingenious strategy to subvert essential activities of GAGs likely prevented host organisms from deleting or inactivating these mechanisms during their evolution. The goal of this review is to provide a mechanistic overview of our current understanding of how microbes subvert GAGs at major steps of pathogenesis, using select GAG-pathogen interactions as representative examples.


title: Bovine viral diarrhoea virus infections and its control. A review.

text: Infections with bovine viral diarrhoea virus continue to plague the cattle industry worldwide. The wish to control the negative effects of the virus has lead to the development of numerous vaccines, but also of eradication schemes. In this paper, a comprehensive overview on BVDV is given: the virus and its clinical manifestations, its occurrence and economic impact, the different routes of transmission, as well as diagnostic methods and objectives. Furthermore, the two major options for BVDV control--eradication and vaccination--are discussed as well as the risk for reintroduction of BVDV after eradication.


Programmed cell death in MHV-induced demyelination.


title: Reassessment of the subcellular localization of p63.

text: p63 is a type II integral membrane protein that has previously been suggested to be a resident protein of a membrane network interposed between the ER and the Golgi apparatus. In the present study, we have produced a polyclonal antibody against the purified human p63 protein to reassess the subcellular distribution of p63 by confocal immunofluorescence, immunoelectron microscopy, and cell fractionation. Double immunofluorescence of COS cells showed significant colocalization of p63 and a KDEL-containing lumenal ER marker protein, except for differences in the staining of the outer nuclear membrane. Immunoelectron microscopy of native HepG2 cells and of COS cells transfected with p63 revealed that both endogenous and overexpressed p63 are predominantly localized in the rough ER. While p63 was colocalized with protein disulfide isomerase, an ER marker protein, very little overlap of p63 was found with ERGIC-53, an established marker for the ER-Golgi intermediate compartment. When rough and smooth membranes were prepared from rat liver, p63 was found to copurify with ribophorin II, a rough ER protein. Both p63 and ribophorin II were predominantly recovered in rough microsomes and were largely separated from the intermediate compartment marker protein p58. From these results it is concluded that p63 is localized in the rough ER.


Infection in solid-organ transplant recipients.


title: Interactions between coronavirus nucleocapsid protein and viral RNAs: implications for viral transcription.

text: The interaction of the mouse hepatitis virus (MHV) nucleocapsid protein (N) and viral RNA was examined. Monoclonal antibody specific for N protein coimmunoprecipitated MHV genomic RNA as well as all six MHV subgenomic mRNAs found in MHV-infected cells. In contrast, monoclonal antibodies to the MHV E2 or E1 envelope glycoproteins, an anti-I-A monoclonal antibody, and serum samples from lupus patients did not immunoprecipitate the MHV mRNAs. Moreover, the anti-N monoclonal antibody did not coimmunoprecipitate vesicular stomatitis virus RNA or host cell RNA under conditions which immunoprecipitated all MHV RNAs. These data suggest a specific interaction between the N protein and the virus-specific mRNAs. Both the membrane-bound and cytosolic small MHV leader-specific RNAs of greater than 65 nucleotides long were immunoprecipitated only by anti-N monoclonal antibody. These data suggest that an N binding site is present within the leader RNA sequences at a site at least 65 nucleotides from the 5' end of genomic RNA and all six subgenomic mRNAs. The larger leader-containing RNAs originating from mRNA 1 and mRNA 6, as well as the MHV negative-stranded RNA, were also immunoprecipitated by the anti-N monoclonal antibody. These data indicate that the MHV N protein is associated with MHV-specific RNAs and RNA intermediates and may play an important functional role during MHV transcription and replication.


title: Human respiratory coronavirus OC43: genetic stability and neuroinvasion.

text: The complete genome sequences of the human coronavirus OC43 (HCoV-OC43) laboratory strain from the American Type Culture Collection (ATCC), and a HCoV-OC43 clinical isolate, designated Paris, were obtained. Both genomes are 30,713 nucleotides long, excluding the poly(A) tail, and only differ by 6 nucleotides. These six mutations are scattered throughout the genome and give rise to only two amino acid substitutions: one in the spike protein gene (I958F) and the other in the nucleocapsid protein gene (V81A). Furthermore, the two variants were shown to reach the central nervous system (CNS) after intranasal inoculation in BALB/c mice, demonstrating neuroinvasive properties. Even though the ATCC strain could penetrate the CNS more effectively than the Paris 2001 isolate, these results suggest that intrinsic neuroinvasive properties already existed for the HCoV-OC43 ATCC human respiratory isolate from the 1960s before it was propagated in newborn mouse brains. It also demonstrates that the molecular structure of HCoV-OC43 is very stable in the environment (the two variants were isolated ca. 40 years apart) despite virus shedding and chances of persistence in the host. The genomes of the two HCoV-OC43 variants display 71, 53.1, and 51.2% identity with those of mouse hepatitis virus A59, severe acute respiratory syndrome human coronavirus Tor2 strain (SARS-HCoV Tor2), and human coronavirus 229E (HCoV-229E), respectively. HCoV-OC43 also possesses well-conserved motifs with regard to the genome sequence of the SARS-HCoV Tor2, especially in open reading frame 1b. These results suggest that HCoV-OC43 and SARS-HCoV may share several important functional properties and that HCoV-OC43 may be used as a model to study the biology of SARS-HCoV without the need for level three biological facilities.


title: Clinical data analysis of 19 cases of community-acquired adenovirus pneumonia in immunocompetent adults.

text: The aim of this study was to investigate the characteristics of clinical manifestations, laboratory tests and imaging changes of community-acquired adenovirus pneumonia in immunocompetent adults. A retrospective study was performed on 19 adult community-acquired adenovirus pneumonia cases in Yantai, whereby the clinical data were collected and analyzed. Of 19 cases, 14 (73.68%) had fever and 17 (89.47%) had cough symptoms. Moreover, 14 cases (73.68%) had normal white blood cell counts, while 11 cases (57.89%) exhibited a reduction in lymphocyte proportion. Among the 19 cases, 17 cases exhibited lesions in a single lung, while 2 cases involved bilateral lungs. The lesions predominantly exhibited ground glass-like changes. The clinical manifestations of adult community-acquired adenovirus pneumonia patients with normal immune functions were mild, with such presenting symptoms as fever, cough, and sputum; most patients did not exhibit high levels of white blood cells or low lymphocyte counts, and the imaging features (ground glass-like effusion) were indicative of single-lung involvement.


Should paramedics intubate patients with SARS-like symptoms?


Development of a critical care triage protocol for pandemic influenza: integrating ethics, evidence and effectiveness.


title: Diagnosis of murine infections in relation to test methods employed.

text: Comparative investigations of Sendai virus, pneumonia virus of mice (PVM), mouse encephalomyelitis virus (mouse polio), minute virus of mice (MVM), and reovirus type 3 (Reo 3) infected murine colonies revealed a 30% higher incidence of positive sera when enzyme-linked immunosorbent assay (ELISA) was employed instead of hemagglutination inhibition (HI) tests. Equivalent sensitivity as in the ELISA was obtained when the same sera were investigated by indirect immunofluorescence (IF) tests. The virus purification techniques described resulted in highly suitable antigens for all indirect ELISA established. Since IIF requires no purified antigens, this test is recommended as an alternative to ELISA as well as to HI and complement fixation (CF) tests for laboratories lacking the necessary equipment for high speed centrifugation. A high incidence of false positive HI reactions was found particularly in Reo 3 routine serology. An updated survey of seromonitoring showed that European murine colonies appeared to be infected far less with Reo 3 if ELISA or IIF tests were employed. During 1982-1984, only 13% of the mouse colonies screened possessed Reo 3 positive sera whereas no natural Reo 3 infection was found in rat colonies. Mouse hepatitis virus (MHV) and the coronaviruses of rats exhibited the highest incidence in murine colonies. A total of 60% of mouse and 41% of rat colonies were found to be infected by these viruses. In comparison with earlier serological surveys, the relative incidence of other murine infections was similar. Antibodies against Bacillus piliformis (Tyzzer's disease) were detected by the IIF test in 41% of the rat colonies screened.


title: Prophylactic fibrinogen administration during complex congenital cardiac surgery leading to thrombosis of a patient's brachial artery and the cardiopulmonary bypass circuit: a case report.

text: Cardiac surgery represents one of the largest consumers of blood products due to the level of blood loss in these patients. As such, there is considerable interest in hemostatic therapies for this patient population. A hemostatic agent that has recently garnered interest is fibrinogen. The conversion of fibrinogen to fibrin by activated thrombin results in clot formation, leading to hemostasis. Currently, a number of studies are underway to evaluate the efficacy of fibrinogen concentrate in cardiac surgery patients. While potentially beneficial in hemostasis for cardiac patients, the current case report provides evidence of the importance of timing in the administration of this product.


Effect of restricted freedom on health in China.


title: Prevalence of Underlying Diseases in Hospitalized Patients with COVID-19: a Systematic Review and Meta-Analysis.

text: Introduction In the beginning of 2020, an unexpected outbreak due to a new corona virus made the headlines all over the world. Exponential growth in the number of those affected makes this virus such a threat. The current meta-analysis aimed to estimate the prevalence of underlying disorders in hospitalized COVID-19 patients. Methods A comprehensive systematic search was performed on PubMed, Scopus, Web of science, and Google scholar, to find articles published until 15 February 2020. All relevant articles that reported clinical characteristics and epidemiological information of hospitalized COVID-19 patients were included in the analysis. Results The data of 76993 patients presented in 10 articles were included in this study. According to the meta-analysis, the pooled prevalence of hypertension, cardiovascular disease, smoking history and diabetes in people infected with SARS-CoV-2 were estimated as 16.37% (95%CI: 10.15%-23.65%), 12.11% (95%CI 4.40%-22.75%), 7.63% (95%CI 3.83%-12.43%) and 7.87% (95%CI 6.57%-9.28%), respectively. Conclusion According to the findings of the present study, hypertension, cardiovascular diseases, diabetes mellitus, smoking, chronic obstructive pulmonary disease (COPD), malignancy, and chronic kidney disease were among the most prevalent underlying diseases among hospitalized COVID-19 patients, respectively.


title: Policy conflicts: gene patents and health care in Canada.

text: Several recent gene patent controversies have energized and refocused the human gene patent debate in Canada. These include the use of the Myriad test for breast cancer by the provinces, patenting of the Severe Acute Respiratory Syndrome virus and a recent Supreme Court decision rejecting the patenting of 'higher life forms'. These cases place the emerging policy conflicts between the innovation and commercialization agenda of the government and the desire to provide equitable access to health care in sharp focus. Another challenge faced by Canada is the powerful influence of the United States in policy decisions. Although these issues have raised awareness about the possibility of reforming the patent system, Parliament has yet to consider any of the suggested reforms of the Canadian patent system and there are no formal proposals pending.


title: Differential regulation of primary and secondary CD8+ T cells in the central nervous system.

text: T cell accumulation and effector function following CNS infection is limited by a paucity of Ag presentation and inhibitory factors characteristic of the CNS environment. Differential susceptibilities of primary and recall CD8+ T cell responses to the inhibitory CNS environment were monitored in naive and CD8+ T cell-immune mice challenged with a neurotropic coronavirus. Accelerated virus clearance and limited spread in immunized mice was associated with a rapid and increased CNS influx of virus-specific secondary CD8+ T cells. CNS-derived secondary CD8+ T cells exhibited increased cytolytic activity and IFN-gamma expression per cell compared with primary CD8+ T cells. However, both Ag-specific primary and secondary CD8+ T cells demonstrated similar contraction rates. Thus, CNS persistence of increased numbers of secondary CD8+ T cells reflected differences in the initial pool size during peak inflammation rather than enhanced survival. Unlike primary CD8+ T cells, persisting secondary CD8+ T cells retained ex vivo cytolytic activity and expressed high levels of IFN-gamma following Ag stimulation. However, both primary and secondary CD8+ T cells exhibited reduced capacity to produce TNF-alpha, differentiating them from effector memory T cells. Activation of primary and secondary CD8+ T cells in the same host using adoptive transfers confirmed similar survival, but enhanced and prolonged effector function of secondary CD8+ T cells in the CNS. These data suggest that an instructional program intrinsic to T cell differentiation, rather than Ag load or factors in the inflamed CNS, prominently regulate CD8+ T cell function.


title: Bariatric surgery in managing diabetes mellitus.

text: PURPOSE OF REVIEW Obesity is a worldwide epidemic, having profound effects on Western populations. Bariatric surgery has long been employed to treat obesity and its related comorbidities. Over time, researchers have amassed significant data to support bariatric surgery in the pursuit of treating diabetes mellitus. This review serves to introduce the most recent findings and their relation to the various bariatric surgical options as bariatric surgery will continue to cement itself in the treatment paradigm of diabetes mellitus. RECENT FINDINGS Numerous studies performed in the past 10 years have demonstrated the improvement or cessation of diabetes with bariatric surgical intervention. In comparing the vertical sleeve gastrectomy and Roux-en-Y gastric bypass, data demonstrate a more beneficial response of diabetes to the Roux-en-Y gastric bypass, and an even further exaggerated response with the biliopancreatic diversion/duodenal switch. The benefit has long been established, but what causes the improvement in diabetes mellitus after bariatric surgery? Recent data suggest a decrease in circulating bile salts as well as changes to inflammatory markers and circulating cytokines. Furthermore, tailoring of existing surgical procedures has led to the development of the SIPS procedure, and its benefit is demonstrated in bypassing a large portion of intestine while eliminating an enteroenterostomy, helping to reduce short gut syndrome and resultant diarrhea. SUMMARY The surgical climate within the bariatric field is changing and will continue to do so in the future. As the understanding of the causes or mechanisms in which bariatric surgery improves metabolic disorders becomes more evident, the process of individualizing care for specific patients will become more prevalent.


title: Pathogenesis of scrapie: agent multiplication in brain at the first and second passage of hamster scrapie in mice.

text: The intracerebral (i.c.) injection of mice with a particular source of hamster passaged scrapie produced disease after an incubation period of 325 +/- 6 days (mean +/- s.e.). The incubation period at the second i.e. passage in mice was reduced to 149 +/- 2 days. Studies were made of the dynamics of agent replication at 1st and 2nd passages in mice. At first passage, there was a 'zero phase' lasting about 175 days, when no infectious agent was detected in brain (or spleen), followed by a period of agent replication which lasted 150 days. At second passage, there was no significant 'zero phase' and agent replication occupied the whole of the incubation period. The occurrence of a 'zero phase' on interspecies passage of scrapie is discussed in relation to other reports of a 'zero phase' in mouse passaged scrapie.


title: Changes in cerebral oxygen saturation and haemoglobin concentration during paediatric cardiac surgery.

text: Although near-infrared spectroscopy (NIRS) enables bedside assessment of cerebral oxygenation, it provides little information on the cause of deoxygenation. The authors aimed to investigate the changes in cerebral oxygenation and haemoglobin concentration and their associations during paediatric cardiac surgery in order to elucidate the physiology underlying cerebral deoxygenation. An observational retrospective study on 399 patients who underwent paediatric cardiac surgery was conducted. With use of NIRS, cerebral oxygen saturation as expressed by tissue oxygen index (TOI) before and after surgery, concentration changes in oxygenated haemoglobin (Δ[HbO2]) and deoxygenated haemoglobin (Δ[HHb]) after surgery were studied as were the associations between these values and clinical variables. TOI decreased after surgery (preoperative versus postoperative value, 66.0% [56.9, 71.3] versus 63.2% [54.3, 69.4], median [25th, 75th percentile], P <0.001) and the decrease was greater in higher category groups in the Risk Adjusted Classification for Congenital Heart Surgery (RACHS-1). [HHb] increased from its baseline (+1.74 μmol/l [-1.57, +5.84], P <0.001) and the increase was greater in higher risk category groups. On the contrary, there was no evidence for a change in [HbO2] (+0.45 μmol/l [-4.76, +5.30], P=0.42). Cerebral oxygen saturation decreased after paediatric cardiac surgery and the decrease was greater in patients of higher risk groups. The increase in [HHb] was considered to play a predominant role in the cerebral deoxygenation noted, in particular in higher RACHS-1 category groups.


Calfhood coronavirus enterocolitis: a clue to the etiology of winter dysentery.


title: Complications of Roux-en-Y gastric bypass and sleeve gastrectomy.

text: Roux-en-Y gastric bypass and sleeve gastrectomy are two of the most common bariatric procedures performed in 2011. Although the complication rates associated with these procedures are low, the consequences of these complications are significant and can be associated with high morbidity and mortality. Timely diagnosis and proper management of these complications are extremely important. The most commonly used radiologic studies in bariatric surgery are the upper GI contrast study and the CT scan, which are used to rule out leak, obstruction, perforation, anastomotic stricture, or pouch dilatation. As with all imaging studies, a negative result should not override strong clinical suspicion of a complication.


title: Detection of porcine teschovirus and enterovirus type II by reverse transcription-polymerase chain reaction.

text: Porcine enteroviruses (PEVs) have been recognized as the causative agents of various clinical manifestations such as fertility disorders, neurological defects, and dermal lesions in pigs. Currently, the diagnosis of PEV infection is carried out by virus isolation, which although useful, is labor- and time-intensive. The present investigation describes the development of a reverse transcription-polymerase chain reaction (RT-PCR) assay for the rapid and sensitive detection of PEVs of cytopathic effect groups I (now known as porcine teschoviruses [PTVs]) and II. The assay described not only detects the PTVs and CPE group II of PEVs but also allows them to be differentiated on the basis of the size of the amplification product, using the same set of oligonucleotide primers. The availability of specific and sensitive molecular diagnostic tools such as the RT-PCR assay described herein should facilitate efficient diagnosis of PTV and CPE group II infections in pigs.


title: A large-volume nebulizer would not be an infectious source for severe acute respiratory syndrome.

text: We attempted to detect the presence of airborne SARS-coronavirus (CoV) in a healthcare setting when a patient with SARS used a humidifier or a large-volume nebulizer (LVN). All of the air samples from the humidifier and LVN were found to have negative SARS-CoV-specific DNA products.


title: Middle East respiratory syndrome coronavirus (MERS-CoV) in dromedary camels, Oman, 2013.

text: A countrywide survey in Oman revealed Middle Eastrespiratory syndrome coronavirus (MERS-CoV) nucleicacid in five of 76 dromedary camels. Camel-derivedMERS-CoV sequences (3,754 nucleotides assembled from partial sequences of the open reading frame (ORF)1a, spike, and ORF4b genes) from Oman and Qatar were slightly different from each other, but closely related to human MERS-CoV sequences from the same geographical areas, suggesting local zoonotic transmission. High viral loads in nasal and conjunctival swabs suggest possible transmission by the respiratory route.


title: CD8+ T cells require perforin to clear West Nile virus from infected neurons.

text: Injury to neurons after West Nile virus (WNV) infection is believed to occur because of viral and host immune-mediated effects. Previously, we demonstrated that CD8+ T cells are required for the resolution of WNV infection in the central nervous system (CNS). CD8+ T cells can control infection by producing antiviral cytokines (e.g., gamma interferon or tumor necrosis factor alpha) or by triggering death of infected cells through perforin- or Fas ligand-dependent pathways. Here, we directly evaluated the role of perforin in controlling infection of a lineage I New York isolate of WNV in mice. A genetic deficiency of perforin molecules resulted in higher viral burden in the CNS and increased mortality after WNV infection. In the few perforin-deficient mice that survived initial challenge, viral persistence was observed in the CNS for several weeks. CD8+ T cells required perforin to control WNV infection as adoptive transfer of WNV-primed wild-type but not perforin-deficient CD8+ T cells greatly reduced infection in the brain and spinal cord and enhanced survival of CD8-deficient mice. Analogous results were obtained when wild-type or perforin-deficient CD8+ T cells were added to congenic primary cortical neuron cultures. Taken together, our data suggest that despite the risk of immunopathogenesis, CD8+ T cells use a perforin-dependent mechanism to clear WNV from infected neurons.


title: Diseases and pathogens associated with mortality in Ontario beef feedlots.

text: This study determined the prevalence of diseases and pathogens associated with mortality or severe morbidity in 72 Ontario beef feedlots in calves that died or were euthanized within 60 days after arrival. Routine pathologic and microbiologic investigations, as well as immunohistochemical staining for detection of bovine viral diarrhea virus (BVDV) antigen, were performed on 99 calves that died or were euthanized within 60 days after arrival. Major disease conditions identified included fibrinosuppurative bronchopneumonia (49%), caseonecrotic bronchopneumonia or arthritis (or both) caused by Mycoplasma bovis (36%), viral respiratory disease (19%), BVDV-related diseases (21%), Histophilus somni myocarditis (8%), ruminal bloat (2%), and miscellaneous diseases (8%). Viral infections identified were BVDV (35%), bovine respiratory syncytial virus (9%), bovine herpesvirus-1 (6%), parainfluenza-3 virus (3%), and bovine coronavirus (2%). Bacteria isolated from the lungs included M. bovis (82%), Mycoplasma arginini (72%), Ureaplasma diversum (25%), Mannheimia haemolytica (27%), Pasteurella multocida (19%), H. somni (14%), and Arcanobacterium pyogenes (19%). Pneumonia was the most frequent cause of mortality of beef calves during the first 2 months after arrival in feedlots, representing 69% of total deaths. The prevalence of caseonecrotic bronchopneumonia caused by M. bovis was similar to that of fibrinosuppurative bronchopneumonia, and together, these diseases were the most common causes of pneumonia and death. M. bovis pneumonia and polyarthritis has emerged as an important cause of mortality in Ontario beef feedlots.


title: High-resolution computed tomography is useful for early diagnosis of severe acute respiratory syndrome-associated coronavirus pneumonia in patients with normal chest radiographs.

text: OBJECTIVE We investigated the usefulness of high-resolution computed tomography (HRCT) in early detection of severe acute respiratory syndrome (SARS)-associated coronavirus pneumonia and analyzed HRCT findings associated with potentially more severe disease. METHODS All patients with suspected SARS and normal chest radiographs on admission within the study period were subjected to HRCT examination. The clinical, radiologic, and laboratory data of patients who were confirmed to have SARS-associated coronavirus infection by a positive nasopharyngeal aspirate, stool or urine reverse transcription-polymerase chain reaction (RT-PCR) and/or serological testing were prospectively followed up and analyzed. Characteristics of patients admitted to the intensive care unit (ICU) and those that were not were compared. RESULTS Among 47 suspected SARS patients with normal chest radiographs, 27 had SARS-associated coronavirus infection confirmed by RT-PCR and/or positive serological testing. Twenty-five of the 27 (93%) patients had HRCT changes compatible with atypical pneumonia, and all 25 patients developed SARS with progressive clinical deterioration. Ten (40%) had unifocal diseases, and 15 had multifocal diseases (60%). Ten (40%) patients had the diseases confined to 1 single lung in the first HRCT, and both lungs were involved at initial presentation in 15 (60%) patients. Concerning the overall extent of the disease at initial presentation, 3 (12%) patients had disease process in all lobes, and the disease was confined to 1 single lobe in 10 (40%) patients. The disease process was mainly peripheral in location (96%), and the lower lobes were more commonly involved (68% in the left lower lobe and 64% in the right lower lobe). Small parapneumonic effusions occurred in 1 (4%) patient. None of the patients with unifocal lesions or single-lung involvement were admitted to the ICU (P < 0.05) (in both comparisons). Subsequent follow-up of the 2 (7%) patients with normal HRCT on admission showed that they were having nonpneumonic SARS-associated coronavirus infection only and were eventually denotified from having SARS. CONCLUSIONS HRCT is useful for early diagnosis of SARS-associated coronavirus pneumonia in patients with normal chest radiographs. HRCT findings in these patients predict potentially severe disease.


title: An infectious recombinant equine arteritis virus expressing green fluorescent protein from its replicase gene.

text: Thus far, systems developed for heterologous gene expression from the genomes of nidoviruses (arteriviruses and coronaviruses) have relied mainly on the translation of foreign genes from subgenomic mRNAs, whose synthesis is a key feature of the nidovirus life cycle. In general, such expression vectors often suffered from relatively low and unpredictable expression levels, as well as genome instability. In an attempt to circumvent these disadvantages, the possibility to express a foreign gene [encoding enhanced green fluorescent protein (eGFP)] from within the nidovirus replicase gene, which encodes two large polyproteins that are processed proteolytically into the non-structural proteins (nsps) required for viral RNA synthesis, has now been explored. A viable recombinant of the arterivirus Equine arteritis virus, EAV-GFP2, was obtained, which contained the eGFP insert at the site specifying the junction between the two most N-proximal replicase-cleavage products, nsp1 and nsp2. EAV-GFP2 replication could be launched by transfection of cells with either in vitro-generated RNA transcripts or a DNA launch plasmid. EAV-GFP2 displayed growth characteristics similar to those of the wild-type virus and was found to maintain the insert stably for at least eight passages. It is proposed that EAV-GFP2 has potential for arterivirus vector development and as a tool in inhibitor screening. It can also be used for fundamental studies into EAV replication, which was illustrated by the fact that the eGFP signal of EAV-GFP2, which largely originated from an eGFP-nsp2 fusion protein, could be used to monitor the formation of the membrane-bound EAV replication complex in real time.


title: Clinical pathology and assessment of pathogen exposure in southern and Alaskan sea otters.

text: The southern sea otter (Enhydra lutris nereis) population in California (USA) and the Alaskan sea otter (E. lutris kenyoni) population in the Aleutian Islands (USA) chain have recently declined. In order to evaluate disease as a contributing factor to the declines, health assessments of these two sea otter populations were conducted by evaluating hematologic and/or serum biochemical values and exposure to six marine and terrestrial pathogens using blood collected during ongoing studies from 1995 through 2000. Samples from 72 free-ranging Alaskan, 78 free-ranging southern, and (for pathogen exposure only) 41 debilitated southern sea otters in rehabilitation facilities were evaluated and compared to investigate regional differences. Serum chemistry and hematology values did not indicate a specific disease process as a cause for the declines. Statistically significant differences were found between free-ranging adult southern and Alaskan population mean serum levels of creatinine kinase, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, calcium, cholesterol, creatinine, glucose, phosphorous, total bilirubin, blood urea nitrogen, and sodium. These were likely due to varying parasite loads, contaminant exposures, and physiologic or nutrition statuses. No free-ranging sea otters had signs of disease at capture, and prevalences of exposure to calicivirus, Brucella spp., and Leptospira spp. were low. The high prevalence (35%) of antibodies to Toxoplasma gondii in free-ranging southern sea otters, lack of antibodies to this parasite in Alaskan sea otters, and the pathogen's propensity to cause mortality in southern sea otters suggests that this parasite may be important to sea otter population dynamics in California but not in Alaska. The evidence for exposure to pathogens of public health importance (e.g., Leptospira spp., T. gondii) in the southern sea otter population, and the naïveté of both populations to other pathogens (e.g., morbillivirus and Coccidiodes immitis) may have important implications for their management and recovery.


title: Emerging zoonotic epidemics in the interconnected global community.

text: The rate at which epidemics of zoonotic disease in humans have surfaced over the past 25 years has shaken--some would say shattered--the assumption that zoonotic diseases are under control, says Paul Gibbs. In this review he analyses the global factors that have led to the increased emergence of zoonotic diseases, sketches several recent epidemics (and where relevant, their relationship to bioterrorism), discusses the lessons learned, and concludes by outlining an agenda for action.


title: Identification of mouse hepatitis virus papain-like proteinase 2 activity.

text: Mouse hepatitis virus (MHV) is a 31-kb positive-strand RNA virus that is replicated in the cytoplasm of infected cells by a viral RNA-dependent RNA polymerase, termed the replicase. The replicase is encoded in the 5'-most 22 kb of the genomic RNA, which is translated to produce a polyprotein of >800 kDa. The replicase polyprotein is extensively processed by viral and perhaps cellular proteinases to give rise to a functional replicase complex. To date, two of the MHV replicase-encoded proteinases, papain-like proteinase 1 (PLP1) and the poliovirus 3C-like proteinase (3CLpro), have been shown to process the replicase polyprotein. In this report, we describe the cloning, expression, and activity of the third MHV proteinase domain, PLP2. We show that PLP2 cleaves a substrate encoding the first predicted membrane-spanning domain (MP1) of the replicase polyprotein. Cleavage of MP1 and release of a 150-kDa intermediate, p150, are likely to be important for embedding the replicase complex in cellular membranes. Using an antiserum (anti-D11) directed against the C terminus of the MP1 domain, we verified that p150 encompasses the MP1 domain and identified a 44-kDa protein (p44) as a processed product of p150. Pulse-chase experiments showed that p150 is rapidly generated in MHV-infected cells and that p44 is processed from the p150 precursor. Protease inhibitor studies revealed that unlike 3CLpro activity, PLP2 activity is not sensitive to cysteine protease inhibitor E64d. Furthermore, coexpression studies using the PLP2 domain and a substrate encoding the MP1 cleavage site showed that PLP2 acts efficiently in trans. Site-directed mutagenesis studies confirmed the identification of cysteine 1715 as a catalytic residue of PLP2. This study is the first to report enzymatic activity of the PLP2 domain and to demonstrate that three distinct viral proteinase activities process the MHV replicase polyprotein.