title: Detection of Australian gill-associated virus (GAV) and lymphoid organ virus (LOV) of Penaeus monodon by RT-nested PCR.

text: A highly sensitive test based on reverse transcription followed by nested polymerase chain reaction (RT-nPCR) was developed to detect the Australian yellow-head-like viruses, gill-associated virus (GAV) and lymphoid organ virus (LOV) of Penaeus monodon. The RT-nPCR detected viral RNA in as little as 10 fg lymphoid organ total RNA isolated from GAV-infected P. monodon. Amplification of serial dilutions of a GAV cDNA clone showed that the nested PCR was sufficiently sensitive to detect a single genome equivalent using a DNA template. The specificity and sensitivity of the RT-nPCR was also demonstrated using experimentally infected P. (Marsupenaeus) japonicus, where GAV sequences could be amplified from lymphoid organ and haemocyte RNA as early as 6 h post infection (p.i.), and from gills by 24 h p.i. In contrast, transmission electron microscopy (TEM) identified nucleocapsids and virions in lymphoid organ cells and haemocytes from Days 3 and 6 p.i., respectively, while there was no evidence of infection in gill cells at any time. The practical application of the RT-nPCR was demonstrated by screening healthy wild-caught P. monodon broodstock. The high prevalence (>98%) of broodstock that were positive by RT-nPCR suggests that LOV is endemic in northern Queensland. In addition, results with lymphoid organ, gill and haemocyte RNA suggest that small gill biopsies may be best suited to the non-sacrificial testing of valuable broodstock. The speed and sensitivity of the RT-nPCR make it a useful adjunct to TEM for diagnosing LOV/GAV infection of P. monodon, with the additional benefit that screening of gill biopsies may facilitate selection of LOV-free broodstock.


title: Outbreak of psittacosis in a group of women exposed to Chlamydia psittaci-infected chickens.

text: Eight cases of psittacosis due to Chlamydia psittaci were identified in May 2013 among 15 individuals involved in chicken gutting activities on a mixed poultry farm in France. All cases were women between 42 and 67 years-old. Cases were diagnosed by serology and PCR of respiratory samples. Appropriate treatment was immediately administered to the eight hospitalised individuals after exposure to birds had been discovered. In the chicken flocks, mainly C. gallinacea was detected, a new member of the family Chlamydiaceae, whereas the ducks were found to harbour predominantly C. psittaci, the classical agent of psittacosis. In addition, C. psittaci was found in the same flock as the chickens that the patients had slaughtered. Both human and C. psittaci-positive avian samples carried the same ompA genotype E/B of C. psittaci, which is widespread among French duck flocks. Repeated grassland rotations between duck and chicken flocks on the farm may explain the presence of C. psittaci in the chickens. Inspection by the veterinary service led to temporary closure of the farm. All birds had to be euthanised on site as no slaughterhouses accepted processing them. Farm buildings and grasslands were cleaned and/or disinfected before the introduction of new poultry birds.


title: Toward Integrated DoD Biosurveillance: Assessment and Opportunities.

text: In the context of the 2012 National Strategy for Biosurveillance, the Office of Management and Budget (OMB) asked the Department of Defense (DoD) to review its biosurveillance programs, prioritize missions and desired outcomes, evaluate how DoD programs contribute to these, and assess the appropriateness and stability of the department's funding system for biosurveillance. DoD sought external analytic support through the RAND Arroyo Center. In response to the questions posed by OMB request, this study finds the following: (1) Current DoD biosurveillance supports three strategic missions. Based mostly on existing statute, the highest-priority mission is force health protection, followed by biological weapons defense and global health security. (2) Guidance issued by the White House on June 27, 2013, specified priorities for planning fiscal year 2015 budgets; it includes an explicit global health security priority, which strengthens the case for this as a key DoD biosurveillance strategic mission. (3) DoD biosurveillance also supports four desired outcomes: early warning and early detection, situational awareness, better decision making at all levels, and forecast of impacts. (4) Programs and measures that address priority missions-force health protection in particular-and desired outcomes should be prioritized over those that do not do so. (5) More near-real-time analysis and better internal and external integration could enhance the performance and value of the biosurveillance enterprise. (6) Improvements are needed in key enablers, including explicit doctrine/policy, efficient organization and governance, and increased staffing and improved facilities for the Armed Forces Health Surveillance Center (AFHSC). (7) AFHSC has requested additional funding to fully implement its current responsibilities under the 2012 Memorandum of Understanding between the Assistant Secretaries of Defense for Health Affairs and for Nuclear, Chemical, and Biological Defense Programs. Additional responsibilities for coordinating the entire DoD biosurveillance enterprise would need concomitant resourcing. (8) There is not a single, unified funding system for the DoD biosurveillance enterprise; the multiple current funding systems would likely benefit from an organizing mechanism with the authority to manage and control funds to meet enterprise goals. Interim guidance issued by the Deputy Secretary of Defense on June 13, 2013, is significant because it is the first policy to explicitly address biosurveillance; it adopts the definition from the National Strategy for Biosurveillance, calls for development of a DoD Directive for biosurveillance, and specifies tasks for DoD's implementation of the Strategy.


title: Antibody titers against bovine coronavirus and shedding of the virus via the respiratory tract in feedlot cattle.

text: OBJECTIVE To describe patterns of seroconversion to bovine coronavirus (BCV) and shedding of BCV from the respiratory tract in feedlot cattle. ANIMALS 1,074 calves in feedlots in Ohio, Texas, and Nebraska. PROCEDURE Nasal swab specimens were obtained at time of arrival (day 0) and at various times during the initial 28 days after arrival at feedlots. Specimens were tested for BCV, using an antigen-capture ELISA. Serum samples were obtained at time of arrival and again 28 days after arrival; sera were analyzed for antibodies to BCV, using an antibody-detection ELISA. RESULTS Samples from 12 groups of cattle entering 7 feedlots during a 3-year period revealed that 78 of 1,074 (7.3%) cattle were shedding BCV (range, 0 to 35.9% within specific groups). At time of arrival, 508 of 814 (62.4%) cattle had low (< 50) or undetectable BCV antibody titers. Seroconversion to BCV during the initial 28 days after arrival was detected in 473 of 814 (58%) cattle tested (range, 20.3 to 84.1 % within specific groups). In cattle shedding BCV from the nasal passages, 49 of 68 (72.1 %) seroconverted, and 472 of 746 (63.3%) cattle that were not shedding the virus seroconverted. CONCLUSIONS AND CLINICAL RELEVANCE Bovine coronavirus can be detected in populations of feedlot cattle in the form of viral shedding as well as seroconversion to the virus. Although only a few cattle were shedding the virus at the time of arrival at a feedlot, most of the cattle seroconverted to BCV by 28 days after arrival.


title: A Randomized Controlled Trial of Anesthesia Guided by Bispectral Index Versus Standard Care: Effects on Cognition.

text: Postoperative cognitive dysfunction, a subtle deterioration of cognitive function after exposure to anesthetics, is reported in 10% to 50% of surgical cases. Delivery of excessive inhalation anesthetics based on minimum alveolar concentration produces greater deep hypnotic times, which may contribute to postoperative cognitive dysfunction. This study tested the impact on cognitive function of balanced anesthetic using electroencephalographic (EEG) guidance vs usual anesthesia. We studied 88 surgical patients: 45 randomly assigned to balanced anesthetic technique with EEG guidance and 43 to standard treatment. Cognitive function was evaluated with the Cambridge Neuropsychological Test Automated Battery-Mild Cognitive Impairment at 3 intervals (preoperatively, 3-5 days postoperatively, and 3-5 months postoperatively). Additionally, 37 age- and sex-matched individuals not undergoing surgery or anesthesia were evaluated at the same intervals. Better outcomes were seen in the intervention group compared with usual care in the short-term/visual memory cognitive domain (P = .02) at 3 to 5 days, but not at 3 to 5 months. Delivery of anesthesia using EEG monitoring systems can reduce cumulative deep hypnotic time without negatively affecting patient physiologic stress, surgical conditions, or cognitive function. Our findings provide data to support optimal anesthetic approaches to improve cognitive function after anesthesia with volatile anesthetics.


title: Laparoscopic Gastric Plication - One Year of Bariatric Surgery in the Emergency County Hospital of Baia Mare.

text: UNLABELLED The main advantages of the laparoscopic gastric plication, in comparison to other bariatric surgery techniques, are the lowest costs and the fewest postopertive complications. This study shows our first year experience with this procedure in the Emergency County Hospital of Baia Mare. METHODS The first 14 laparoscopic gastric pulications performed in 2014 were studied retrospectively: there were 13 women and one man, with age ranging from 25 to 60 years old and a body mass index (BMI) from 35,7 to 59,8. Postoperative, the patients' evolution was evaluated 9 days, 1 month, 3 months, 6 months, 1 year after the surgery and before writing this paper, taking into account excess weight loss percentage and comorbidities' progress. RESULTS The average weight loss was 21,42 kg and the weight loss percentage was 36,52%. There were no postoperative complications in need of surgical reintervention. CONCLUSIONS The results were very good, the costs were low and the postoperative complications had a low rate. Therefore, there is a recommendation on performing this surgery in every hospital equipped for laparoscopic surgery.


title: [Preventive and therapeutic effects of recombinant IFN-alpha2b nasal spray on SARS-CoV infection in Macaca mulata].

text: OBJECTIVE To study the preventive and therapeutic effects of recombinant IFN-alpha2b for nasal spray on SARS-CoV infection in Macaca mulata (rhesus monkey). METHODS Ten rhesus monkeys were divided into two groups, 5 in interferon group, and 5 in control group. Before and after SARS-CoV attack, the virus was detected in samples such as pharyngeal swab in all the two groups by Real-time PCR (RT-PCR) and virus isolation was performed. RESULTS After virus attack, the level of SARS-CoV-specific IgG and neutralizing antibody were induced by SARS-CoV in the interferon group was weaker than in control group. Hematology items showed no apparent changes after virus attack in treated group. Through pathological examination, the morphology of the lung tissues of two Macaques in the treated group was normal, while the other three displayed the interstitial pneumonia with the thickened septum and infiltration with mononuclear cells. Among which, one monkey showed part of thickened septum fused with each other. These lesions in the interferon treated animals were similar to those seen in the animals in control group, but with smaller scope of pathological changes. No significant abnormity was detected in other organs. CONCLUSION Recombinant IFN-alpha2b could effectively interdict or weaken SARS-CoV injury in monkeys.


title: Management of ST-Elevation Myocardial Infarction in the COVID-19 Era: The Role of Thrombosis and Anticoagulation Strategy.

text: Cardiac manifestations of COVID-19 include myocarditis, demand ischemia, myocardial infarction and arrhythmias with prothrombotic state being a major underlying pathogenetic mechanism. In this report we present a case of a 57-year-old, otherwise healthy, woman who presented with chest pain and nausea and was found to have an inferior wall ST-elevation myocardial infarction (STEMI) in the setting of an active COVID-19 infection. Angiography revealed tortuous coronary arteries with a 100% right coronary artery occlusion with high thrombus burden and normal left coronary system. In light of the available literature regarding the pro-thrombotic effects of this novel corona virus, we continued full dose anticoagulation with Enoxaparin after the cardiac catheterization and transitioned to rivaroxaban and we also continued the patient on dual antiplatelet therapy prior to discharge.


title: Evaluation of a canine small intestinal submucosal xenograft and polypropylene mesh as bioscaffolds in an abdominal full-thickness resection model of growing rats.

text: We evaluated the biological scaffold properties of canine small intestinal submucosa (SIS) compared to a those of polypropylene mesh in growing rats with full-thickness abdominal defects. SIS is used to repair musculoskeletal tissue while promoting cell migration and supporting tissue regeneration. Polypropylene mesh is a non-resorbable synthetic material that can endure mechanical tension. Canine SIS was obtained from donor German shepherds, and its porous collagen fiber structure was identified using scanning electron microscopy (SEM). A 2.50-cm(2) section of canine SIS (SIS group) or mesh (mesh group) was implanted in Sprague-Dawley rats. At 1, 2, 4, 12, and 24 weeks after surgery, the implants were histopathologically examined and tensile load was tested. One month after surgery, CD68+ macrophage numbers in the SIS group were increased, but the number of CD8+ T cells in this group declined more rapidly than that in rats treated with the mesh. In the SIS group, few adhesions and well-developed autologous abdominal muscle infiltration into the SIS collagen fibers were observed. No significant differences in the tensile load test results were found between the SIS and mesh groups at 24 weeks. Canine SIS may therefore be a suitable replacement for artificial biological scaffolds in small animals.


title: Human catalase, its polymorphisms, regulation and changes of its activity in different diseases.

text: Catalase (CAT) is a well-studied enzyme that plays an important role in protecting cells against the toxic effects of hydrogen peroxide. In human, it has been implicated in different physiological and pathological conditions. This review summarizes the information available on the function and role of CAT polymorphisms in pathogenesis of various pathophysiological states as well as on the regulation of CAT gene expression. Numerous studies have described the CAT polymorphisms and their link with various diseases. Changes in the CAT levels were reported in many different diseases and polymorphisms in the CAT gene were shown to be associated with different pathophysiological states, e.g. hypertension, diabetes mellitus, insulin resistance, dyslipidaemia, asthma, bone metabolism or vitiligo. Regulation of the CAT gene expression plays an important role in the levels of CAT. The catalase gene expression is regulated by various mechanisms involving e.g. peroxisome proliferator-activated receptor γ (PPARγ), tumour necrosis factor α (TNF-α), p53 protein and hypermethylation of CpG islands in the catalase promoter. Transcription of the CAT gene is mainly influenced by the -262 C/T and -844 A/G polymorphisms. A common polymorphism -262 C/T in the promoter region has been found to be associated with altered CAT activities. Apart from genetic factors, the activities of CAT may be affected by age, seasonal variations, physical activity, or a number of chemical compounds. Future investigations are necessary to elucidate the role of CAT in pathogenesis of oxidative stress-related diseases.


title: Heparan sulfate is a selective attachment factor for the avian coronavirus infectious bronchitis virus Beaudette.

text: The avian coronavirus infectious bronchitis virus (IBV) strain Beaudette is an embryo-adapted virus that has extended species tropism in cell culture. In order to understand the acquired tropism of the Beaudette strain, we compared the S protein sequences of several IBV strains. The Beaudette strain was found to contain a putative heparan sulfate (HS)-binding site, indicating that the Beaudette virus may use HS as a selective receptor. To ascertain the requirements of cell-surface HS for Beaudette infectivity, we assayed for infectivity in the presence of soluble heparin as a competitor and determined infectivity in mutant cell lines with no HS or glycosaminoglycan expression. Our results indicate that HS plays a role as an attachment factor for IBV, working in concert with other factors like sialic acid to mediate virus binding to cells, and may explain in part the extended tropism of IBV Beaudette.


title: The cost of seasonal respiratory illnesses in Australian children: the dominance of patient and family costs and implications for vaccine use.

text: Respiratory viral infections are one of the next group of diseases likely to be targeted for prevention in childhood by the use of vaccines. To begin collecting necessary epidemiology and cost information about the illnesses caused by these viruses, we conducted a prospective cohort study in 118 Melbourne children between 12 and 71 months of age during winter and spring 2001. We were interested in calculating an average cost per episode of community-managed acute respiratory disease, in identifying the key cost drivers of such illness, and to identify the proportion of costs borne by the patient and family. There were 202 community-managed influenza-like illnesses identified between July and December 2001, generating 89 general practitioner visits, and 42 antibiotic prescriptions. The average cost of community-managed episodes (without hospitalisation) was dollar 241 (95% CI dollar 191 to dollar 291), with the key cost drivers being carer time away from usual activities caring for the ill child (70% of costs), use of non-prescription medications (5.4%), and general practice visits (5.0%). The patient and family met 87 per cent of total costs. The lowest average cost occurred in households from the highest income bracket. Acute respiratory illness managed in the community is common, with the responsibility for meeting the cost of episodes predominantly borne by the patient and family in the form of lost productivity. These findings have implications for preventive strategies in children, such as the individual use of, or implementation of public programs using, currently available vaccines against influenza and vaccines under development against other viral respiratory pathogens.


title: The molecular basis of antibody-mediated neutralization of West Nile virus.

text: The study of the interaction between the West Nile virus envelope protein and monoclonal antibodies has provided insight into the molecular mechanisms of neutralization. Structural studies have identified an epitope on the lateral ridge of domain III of the West Nile virus E protein that is recognized by antibodies with the strongest neutralizing activity in vitro and in vivo. Antibodies that bind to this epitope are particularly inhibitory because they block infection at a post-attachment step and at concentrations that result in a low occupancy of the available sites on the virion.


title: A case control study of potential enteric pathogens for calves raised in cow-calf herds.

text: A matched case control study was performed to describe the epidemiological features of potential enteric pathogens for calves reared in 53 cow-calf herds located in western Switzerland. A total of 106 diarrhoeic calves and 126 healthy control calves were collected, all calves were less than 4 months old. Faecal samples were analysed for presence of infectious agents related to calf diarrhoea including enterotoxigenic E. coli, Verotoxin producing E. coli (VTEC), Campylobacter sp., Yersinia sp., Salmonella sp., rotavirus, coronavirus, helminths and coccidian protozoa. Multivariate logistic models were used to analyse the relationship between presence of infection and onset of diarrhoea. The study provided evidence of significant associations between diarrhoea and infection with rotavirus, Campylobacter coli and the presence of Verotoxin in faecal samples. With the exception of Cryptosporidium parvum intestinal parasites including Strongylidae and Eimeria sp. were found to be less prevalent in cases than in controls. Control calves were significantly more frequently infected with Strongyloides papillosus than case animals.


title: A near-fatal infection with oseltamivir-resistant seasonal influenza A in a previously healthy child: Case report.

text: A case of near-fatal oseltamivir-resistant seasonal influenza A infection in a previously healthy four-year-old boy is reported. This case highlights three important points for physicians: oseltamivir-resistant influenza A (H1N1) has recently emerged in North America; contrary to previously held beliefs, such strains are capable of causing severe disease in healthy children; and given this change in epidemiology, clinicians caring for children with severe seasonal influenza A infection should consider empiric dual therapy with oseltamivir and amantadine.


title: Essential function in vivo for Dicer-2 in host defense against RNA viruses in drosophila.

text: The fruit fly Drosophila melanogaster is a model system for studying innate immunity, including antiviral host defense. Infection with drosophila C virus triggers a transcriptional response that is dependent in part on the Jak kinase Hopscotch. Here we show that successful infection and killing of drosophila with the insect nodavirus flock house virus was strictly dependent on expression of the viral protein B2, a potent inhibitor of processing of double-stranded RNA mediated by the essential RNA interference factor Dicer. Conversely, flies with a loss-of-function mutation in the gene encoding Dicer-2 (Dcr-2) showed enhanced susceptibility to infection by flock house virus, drosophila C virus and Sindbis virus, members of three different families of RNA viruses. These data demonstrate the importance of RNA interference for controlling virus replication in vivo and establish Dcr-2 as a host susceptibility locus for virus infections.


title: Zika Virus: A Basic Overview of an Emerging Arboviral Infection in the Western Hemisphere.

text: Since February 2015, Zika virus has spread throughout the Western Hemisphere, starting in Brazil. As of March 2016, autochthonous transmission has been reported in at least 31 countries or territories. For countries in the Americas, the spread of Zika virus, a previously unfamiliar disease, follows similar emerging infection introductions of West Nile virus and Chikungunya virus and their spread throughout the American continents and the Caribbean nations. The Pan American Health Organization and the World Health Organization have issued alerts and a Public Health Emergency of International Concern announcement related to the recent cluster of microcephaly cases and other neurological disorders in Brazil that are temporally associated with Zika virus, which highlights the possible adverse impact of viral infection. This article provides an overview of the Zika virus infection and presents the historical background of the virus, a description of the pathogen, the epidemiology and clinical spectrum of Zika virus infection, diagnosis and treatment approaches, and prevention and control measures. Understanding what is known about the virus and its clinical presentation will assist in prevention, detection, and response measures to reduce and control the spread of the virus throughout the Western Hemisphere. (Disaster Med Public Health Preparedness. 2016;page 1 of 6).


SARS. China dumps CDC head, probes lab.


title: Antibody-dependent enhancement of feline infectious peritonitis virus infection in feline alveolar macrophages and human monocyte cell line U937 by serum of cats experimentally or naturally infected with feline coronavirus.

text: Infection of the type II feline infectious peritonitis virus (FIPV) strain 79-1146 to primary feline alveolar macrophages and human monocyte cell line U937 was enhanced by the sera of cats experimentally infected with the 79-1146 strain, but not those of cats infected with KU-2 or UCD-1 strain of type I FIPV. The experiments using sera of cats with feline infectious peritonitis (FIP) and of cats naturally infected with feline coronavirus (FCoV) revealed that infection of the FIPV 79-1146 strain to the U937 cells was enhanced only by the sera of cats infected with type II FIPV or feline enteric coronavirus. The samples positive for antibody-dependent enhancement (ADE) activity had high neutralizing antibody titers against the FIPV 79-1146 strain and the samples negative for ADE activity had low neutralizing antibody titers. These findings support the previous results where a monoclonal antibody with neutralizing activity had high ADE activity, suggesting that there was a close relationship between the neutralization and enhancement sites. And then it is also suggested that ADE of infection is likely to be induced by re-infection with the same serotype of virus in type II FIPV infection. Furthermore, U937 cells are considered useful and can be substituted for the feline macrophages for determining ADE of FIPV-infection.


title: Physiology and behavior of dogs during air transport.

text: Twenty-four beagles were used to measure physiological and behavioral reactions to air transport. Each of 3 groups of 4 sedated (with 0.5 mg/kg body weight of acepromazine maleate) and 4 non-sedated (control) dogs was flown on a separate flight between Montreal, Quebec, and Toronto, Ontario, after being transported by road from Quebec City to Montreal. Saliva and blood samples were taken before ground and air transport and after air transport. The heart rate was monitored during the whole experiment except during ground transport, and behavior was monitored by video during air transport. Sedation did not affect any of the variables measured. The mean plasma cortisol concentration was significantly higher (P < 0.05) after ground transport than at baseline (225.3 vs 134.5 nmol/L); the mean salivary cortisol concentration was significantly higher (P < 0.05) after both ground and air transport than at baseline (16.2 and 14.8, respectively, vs 12.6 nmol/L). The mean neutrophil count was significantly higher (P < 0.05) after both ground and air transport than at baseline (80.6 and 81.4, respectively, vs 69.5 per 100 white blood cells), whereas the mean lymphocyte count was significantly lower (P < 0.05) (13.2 and 13.7, respectively, vs 22.4 per 100 white blood cells). Loading and unloading procedures caused the largest increase in heart rate. On average, the dogs spent more than 50% of the time lying down, and they remained inactive for approximately 75% of the time, except during take-off. These results suggest that transportation is stressful for dogs and that sedation with acepromazine, at the dosage and timing used, does not affect the physiological and behavioral stress responses of dogs to air transport.


title: Tropical sprue: revisited.

text: OBJECTIVE To review the experience of patients presenting with clinical manifestations of tropical sprue and assess their diagnosis and management, response to treatment and follow up. METHODS This single center retrospective descriptive study was done at Shifa International Hospital, Islamabad from January 1994 to January 2003. All patients who presented with diarrhea, anorexia, weight loss and anemia and who proved to have partial villous atrophy on intestinal biopsy and had responded to treatment with antibiotic and folic acid,were included in this review. RESULTS A total of 42 patients were encountered during these years. There were 31 (74.0%) males and 11 (26%) females. The age ranged from 17-66 years. All patients presented with diarrhea, weight loss, anorexia and had megaloblastic anemia. In all patients, a distal duodenal biopsy showed partial villous atrophy. All were treated with tetracycline 1 g per day and folic acid 5 mg per day and responded to treatment within 4 weeks. Total treatment lasted 3 months and resulted in complete resolution of symptoms and gain of weight. The follow up lasted for a mean of 5 years and no relapses were noted. CONCLUSION Tropical sprue presents with diarrhea, anorexia, weight loss, and megaloblastic anemia. The partial villous atrophy has been a constant finding. The response to treatment to tetracycline and folic acid has been uniformly successful.


title: E. coli multiresistant meningitis after transrectal prostate biopsy.

text: Escherichia coli meningitis is a frequent pathology in children younger than 3 years old, but is an uncommon disease in adults. E. coli infection is the main cause of intrahospital bacteremia as a consequence of the employment of different medical procedures. Our patient, male, 69 years old, presented with fever, progressive difficulty in breathing, and shivers 24 h after transrectal prostate biopsy, with an absence of any other symptoms. He received prophylactic treatment with ciprofloxacin and later empirical treatment with ampicillin and tobramicin. After that, the patient presented with fever, headache, behavioral changes, somnolence, disorientation, a fluctuating level of conscience, cutaneous widespread pallor, and acute urinary retention. On physical exploration, we observed generalized hypoventilation, Glasgow 10, stiffness of the neck, inconclusive Kernig; the remaining neurological exploration was normal. Systematic of blood: leukocytes = 8,510/mm3 (94.5% polymorphonuclear), platelet = 87,000/mm3, pH = 7.51, pCO2 = 28.8 mmHg, pO2 = 61 mmHg, O2 saturation = 93.8%, and remaining values were normal. Chest X- ray, cranial CT scan, urine cultures were normal. Blood culture: E. coli. CSF: glucose <0.4 g/l, total proteins = 3.05 g/l, PMN = 7 cells. Microscopic examination of the CSF: Gram-negative bacilli; CSF's culture: abundant E. coli. The case of acute meningitis by multiresistant E. coli after transrectal prostate biopsy presented demonstrates that antibiotic prevention with ciprofloxacin is not absolutely risk free. Besides the use of antibiotic prevention for multiresistant microorganisms, the urologist and other physicians involved in the procedure must not forget that the rate of major complications of transrectal prostate biopsy is 1%, especially when it is performed in patients who will not benefit from that biopsy.


title: [Analysis on diagnosis, treatment and influential factors in fever patients during epidemic of severe acute respiratory syndrome].

text: OBJECTIVE To analyze the etiology, therapy and influential factors in fever patients during the epidemic of severe acute respiratory syndrome (SARS), so as to define early and differential diagnosis for SARS from other disease with fever. METHODS The epidemiological and the clinical data of 4060 patients with fever admitted to the emergency department of the Third Hospital affiliated to Peking University from March 5th to May 10th in 2003 were analyzed, and their influential factors were evaluated. RESULTS Of 4060 patients with fever 275 cases were admitted to the isolated ward, 145 cases were diagnosed as SARS (3.75 percent), 29 belonged to suspected cases, and in 101 cases SARS was ruled out. Ninety-three cases were non-SARS-pneumonia, 2 were epidemic encephalitis, 8 were epidemic cerebrospinal meningitis, and 3 cases were epidemic hemorrhagic fever. CONCLUSION During epidemic of SARS, the etiology of fever is influenza or other virus infection in 90 percent of patients. Non-SARS-pneumonia and infectious diseases with fever accounted for certain proportion of patients with fever. Our diagnostic level should be improved. SARS could not be ruled out only by a high white blood cells count. Chest X-ray examination have to be repeated, because abnormal chest X-ray do not always appear at the same time with fever.


title: Neutralization determinants of laboratory strains and field isolates of equine arteritis virus: identification of four neutralization sites in the amino-terminal ectodomain of the G(L) envelope glycoprotein.

text: The N-terminal hydrophilic ectodomain of the G(L) envelope glycoprotein of equine arteritis virus (EAV) contains neutralization determinants of the virus. We developed a panel of 17 neutralizing murine monoclonal antibodies (MAbs) to further characterize the neutralization determinants of EAV. Included were 6 MAbs previously raised against a laboratory strain (EAVUCD) of the original Bucyrus strain of EAV, as well as 11 additional MAbs that were raised against a neutralization-resistant variant [escape mutant (EM)] virus (EM6D10) that was derived from EAVUCD. All MAbs raised against EAVUCD and 4 of the MAbs raised against EM6D10 (2B3, 5F8, 8D4, and 10B4) reacted with the corresponding G(L) envelope glycoprotein in a Western immunoblotting assay, whereas the remaining 7 MAbs raised against EM6D10 did not react with any viral protein in the immunoblotting assay but competitively inhibited the binding of MAbs 2B3, 5F8, 8D4, and 10B4, indicating that they also recognize epitopes on the G(L) protein. A panel of 18 EM viruses raised to the MAb panel, 19 field isolates of EAV from North America and Europe, the modified-live virus vaccine (ARVAC), and 3 other laboratory strains of EAV were characterized by microneutralization assay with the panel of neutralizing MAbs and polyclonal rabbit and horse antisera. Comparative analysis of the nucleotide sequences of ORF5 and the deduced amino acid sequences of the G(L) protein of individual EM viruses and field isolates of EAV identified four distinct neutralization sites. These sites include amino acids 49 (site A), 61 (site B), 67 through 90 (site C), and 99 through 106 (site D). With the notable exception of site A, the sites were all located in the V1 variable region (amino acids 61-121) within the second half of the N-terminal hydrophilic ectodomain of the G(L) protein. Site D includes several overlapping linear epitopes which appear to interact with amino acids in the other three sites to form conformationally dependent epitopes. Amino acid substitutions within any of these four sites can alter the neutralization phenotype of individual strains of EAV.


title: Inhibition of severe acute respiratory syndrome virus replication by small interfering RNAs in mammalian cells.

text: Severe acute respiratory syndrome (SARS) is an acute respiratory infectious disease that spread worldwide in early 2003. The cause was determined as a novel coronavirus (CoV), SARS-associated CoV (SARS-CoV), with a single-stranded, plus-sense RNA. To date, no effective specific treatment has been identified. To exploit the possibility of using RNA interference as a therapeutic approach to fight the disease, plasmid-mediated small interfering RNAs (siRNAs) were generated to target the SARS-CoV genome. The expression of siRNAs from two plasmids, which specifically target the viral RNA polymerase, effectively blocked the cytopathic effects of SARS-CoV on Vero cells. These two plasmids also inhibited viral replication as shown by titer assays and by an examination of viral RNA and protein levels. Thus, our results demonstrated the feasibility of developing siRNAs as effective anti-SARS drugs.


title: SARS in Singapore--predictors of disease severity.

text: INTRODUCTION Severe acute respiratory syndrome (SARS) affected 8096 individuals in 29 countries, with 774 deaths. In Singapore, there were 238 cases of SARS with 33 deaths. A retrospective analysis was performed to identify predictors of poor outcome in patients with SARS locally. MATERIALS AND METHODS Clinical, laboratory and outcome data of 234 patients admitted to Tan Tock Seng Hospital and Singapore General Hospital were collected and analysed. Only data collected at the time of admission were used in the analysis for predictors of poor outcome. Adverse events were defined as admission to the intensive care unit or death. RESULTS Clinical (temperature, FiO2) and laboratory [leukocyte, lymphocyte, neutrophil, platelet, lactate dehydrogenase (LDH), albumin] trends in groups with and without an adversarial event were presented. Fifty patients experienced an adverse event. On univariate analysis, male gender, advanced age, presence of comorbidities, neutrophilia, lymphopaenia, hyponatraemia, hypoalbuminaemia, transaminitis and elevated LDH or C-reactive protein were found to be significant predictors. On multivariate analysis, predictors of poor outcome were increased age [odds ratio (OR) 1.73 for every 10-year increase; 95% CI, 1.35 to 2.21], neutrophilia (OR 1.06 for every 1 x 10(9)/L increase; 95% CI, 1.02 to 1.11) and high LDH (OR 1.17 for every 100 U/L increase; 95% CI, 1.02 to 1.34). None of the 12 paediatric patients had an adverse event. CONCLUSION Advanced age, neutrophilia and high LDH predict poor outcomes in patients with SARS.


title: Investigating turkey enteric coronavirus circulating in the Southeastern United States and Arkansas during 2012 and 2013.

text: Periodic monitoring of poultry flocks in the United States via molecular diagnostic methods has revealed a number of potential enteric viral pathogens in continuous circulation in turkeys and chickens. Recently turkey integrators in the Southeastern United States and Arkansas experienced an outbreak of moderate to severe enteritis associated with turkey enteric coronavirus (TCoV), and numerous enteric samples collected from turkey flocks in these areas tested positive for TCoV via real-time reverse-transcriptase PCR (RRT-PCR). This report details the subsequent sequence and phylogenetic analysis of the TCoV spike glycoprotein and the comparison of outbreak-associated isolates to sequences in the public database. TCoVs investigated during the present outbreak grouped geographically based upon state of origin, and the RRT-PCR assay was a good indicator of subsequent seroconversion by TCoV-positive turkey flocks.


title: The psychological impact of severe acute respiratory syndrome outbreak on healthcare workers in emergency departments and how they cope.

text: OBJECTIVES The objectives of the present study were to examine the degree and the sources of mental distress and the coping strategies adopted by healthcare workers (HCW) of emergency departments (ED) in Hong Kong during the outbreak of severe acute respiratory syndrome (SARS). METHODS Questionnaires were sent to all doctors, nurses and healthcare assistants (HCA) working in the ED of all public hospitals. The overall degree of mental distress was measured by a single-item 11-point Likert scale. The source of distress was measured by an 18-item questionnaire, which was designed based on the experience of clinical psychologist colleagues providing counselling to staff taking care of SARS patients. The Brief Cope questionnaire was used to study coping strategies adopted by staff. RESULTS A total of 1260 questionnaires were sent out and the response rate was approximately 37%. The mean overall distress level was 6.19 out of a 10-point scale. The mean overall distress levels for doctors, nurses and HCA were 5.91, 6.52 and 5.44, respectively (F(2,420)=6.47, P<0.005). The overall distress level for nurses was significantly higher than for HCA (P<0.005) but not doctors. The overall distress level was highly and significantly correlated with the six sources of distress: vulnerability/loss of control (r=0.68); health of self (r=0.62); spread of virus (r=0.60); health of family and others (r=0.59); changes in work (r=0.46); being isolated (r=0.45). The scores for nurses were significantly higher than for doctors in terms of the six sources of distress (all P values <0.01). HCA were significantly higher than doctors (but not nurses) in worrying about their family's and others' health (P<0.05). In terms of coping strategies, doctors were significantly more likely than nurses and HCA to use planning (P<0.05 and <0.01 respectively); nurses were significantly more likely than doctors to use behavioural disengagement (P<0.01); whereas HCA were significantly more likely than doctors to use self distractions (P<0.05). CONCLUSIONS SARS had caused a significant level of distress among ED staff. The distress level was highest for nurses, followed by doctors and HCA. The three most important variables that could account for the distress level were loss of control/vulnerability, fear for self-health and spread of the virus. Overall, the more frequently adopted coping strategies were acceptance, active coping, and positive framing.


title: Detection of rotaviruses and intestinal lesions in broiler chicks from flocks with runting and stunting syndrome (RSS).

text: The intestinal tract and intestinal contents were collected from 34 stunted, 5-to-14-day-old broiler chicks from eight flocks with runting and stunting syndrome (RSS) in Northern Germany to investigate intestinal lesions and the presence of enteric pathogens with a special focus on rotaviruses (RVs). Seven chicks from a healthy flock were used as controls. Severe villous atrophy was seen in chicks from six flocks with RSS but not in the control flock. Lesions were often "regionally" distributed in the middle-to-distal small intestine. Transmission electron microscopy (TEM), polyacrylamide-gel electrophoresis (PAGE), reverse-transcriptase polymerase chain reaction (RT-PCR), and seminested RT-PCR were used for detection and characterization of RVs. The PAGE allows discrimination of different RV groups, and the RT-PCR was used to verify the presence of group (gp) A RVs. RVs were detected (by all methods) in 32 of 34 chicks from the flocks with RSS. By TEM (negative staining), RV particles were observed in intestinal contents of 28 chicks from the flocks with RSS. PAGE analysis showed four RV groups: gpA, gpD, gpF, and gpG. Group A RVs were detected in four chicks from two flocks with RSS, without intestinal lesions. GpD RVs were detected in 12 chicks of five flocks with RSS, 10 of them with severe villous atrophy. GpF RVs were confirmed in four chicks from three flocks with RSS and in two birds in the control flock. GpG RVs were verified in two chicks from two flocks with RSS, one with, and one without, intestinal lesions. At present, PCR methods are only available for detection of gpA RVs. Using RT-PCR, gpA RVs were identified in samples from 22 chicks including samples of two chicks from the control flock. Statistical analysis revealed a positive correlation between presence of gpD RV and severe villous atrophy in flocks with RSS. The results suggest that gpD RV plays a major role in the pathogenesis of RSS.


title: Seroprevalence of IgG antibodies to SARS-coronavirus in asymptomatic or subclinical population groups.

text: We systematically reviewed the current understanding of human population immunity against SARS-CoV in different groups, settings and geography. Our meta-analysis, which included all identified studies except those on wild animal handlers, yielded an overall seroprevalence of 0.10% [95% confidence interval (CI) 0.02-0.18]. Health-care workers and others who had close contact with SARS patients had a slightly higher degree of seroconversion (0.23%, 95% CI 0.02-0.45) compared to healthy blood donors, others from the general community or non-SARS patients recruited from the health-care setting (0.16%, 95% CI 0-0.37). When analysed by the two broad classes of testing procedures, it is clear that serial confirmatory test protocols resulted in a much lower estimate (0.050%, 95% CI 0-0.15) than single test protocols (0.20%, 95% CI 0.06-0.34). Potential epidemiological and laboratory pitfalls are also discussed as they may give rise to false or inconsistent results in measuring the seroprevalence of IgG antibodies to SARS-CoV.


title: Role of the coronavirus E viroporin protein transmembrane domain in virus assembly.

text: Coronavirus envelope (E) proteins are small (approximately 75- to 110-amino-acid) membrane proteins that have a short hydrophilic amino terminus, a relatively long hydrophobic membrane domain, and a long hydrophilic carboxy-terminal domain. The protein is a minor virion structural component that plays an important, not fully understood role in virus production. It was recently demonstrated that the protein forms ion channels. We investigated the importance of the hydrophobic domain of the mouse hepatitis coronavirus (MHV) A59 E protein. Alanine scanning insertion mutagenesis was used to examine the effect of disruption of the domain on virus production in the context of the virus genome by using a MHV A59 infectious clone. Mutant viruses exhibited smaller plaque phenotypes, and virus production was significantly crippled. Analysis of recovered viruses suggested that the structure of the presumed alpha-helical structure and positioning of polar hydrophilic residues within the predicted transmembrane domain are important for virus production. Generation of viruses with restored wild-type helical pitch resulted in increased virus production, but some exhibited decreased virus release. Viruses with the restored helical pitch were more sensitive to treatment with the ion channel inhibitor hexamethylene amiloride than were the more crippled parental viruses with the single alanine insertions, suggesting that disruption of the transmembrane domain affects the functional activity of the protein. Overall the results indicate that the transmembrane domain plays a crucial role during biogenesis of virions.


title: Replication and packaging of coronavirus infectious bronchitis virus defective RNAs lacking a long open reading frame.

text: The construction of a full-length clone of the avian coronavirus infectious bronchitis virus (IBV) defective RNA (D-RNA), CD-91 (9,080 nucleotides [Z. Penzes et al., Virology 203:286-293]), downstream of the bacteriophage T7 promoter is described. Electroporation of in vitro T7-transcribed CD-91 RNA into IBV helper virus-infected primary chick kidney cells resulted in the production of CD-91 RNA as a replicating D-RNA in subsequent passages. Three CD-91 deletion mutants were constructed--CD-44, CD-58, and CD-61--in which 4,639, 3,236, and 2,953 nucleotides, respectively, were removed from CD-91, resulting in the truncation of the CD-91 long open reading frame (ORF) from 6,465 to 1,311, 1,263, or 2,997 nucleotides in CD-44, CD-58, or CD-61, respectively. Electroporation of in vitro T7-transcribed RNA from the three constructs into IBV helper virus-infected cells resulted in the replication and packaging of CD-58 and CD-61 but not CD-44 RNA. The ORF of CD-61 was further truncated by the insertion of stop codons into the CD-61 sequence by PCR mutagenesis, resulting in constructs CD-61T11 (ORF: nucleotides 996 to 1,058, encoding 20 amino acids), CD-61T22 (ORF: nucleotides 996 to 2,294, encoding 432 amino acids), and CD-61T24 (ORF: nucleotides 996 to 2,450, encoding 484 amino acids), all of which were replicated and packaged to the same levels as observed for either CD-61 or CD-91. Analysis of the D-RNAs showed that the CD-91- or CD-61-specific long ORFs had not been restored. Our data indicate that IBV D-RNAs based on the natural D-RNA, CD-91, do not require a long ORF for efficient replication. In addition, a 1.4-kb sequence, corresponding to IBV sequence at the 5' end of the 1b gene, may be involved in the packaging of IBV D-RNAs or form part of a cis-acting replication element.


title: Pandemic influenza and critical infrastructure dependencies: possible impact on hospitals.

text: Hospitals will be particularly challenged when pandemic influenza spreads. Within the health sector in general, existing pandemic plans focus on health interventions to control outbreaks. The critical relationship between the health sector and other sectors is not well understood and addressed. Hospitals depend on critical infrastructure external to the organisation itself. Existing plans do not adequately consider the complexity and interdependency of systems upon which hospitals rely. The failure of one such system can trigger a failure of another, causing cascading breakdowns. Health is only one of the many systems that struggle at maximum capacity during "normal" times, as current business models operate with no or minimal "excess" staff and have become irreducible operations. This makes interconnected systems highly vulnerable to acute disruptions, such as a pandemic. Companies use continuity plans and highly regulated business continuity management to overcome process interruptions. This methodology can be applied to hospitals to minimise the impact of a pandemic.


title: Acute and persistent infection of human neural cell lines by human coronavirus OC43.

text: Human coronaviruses (HuCV) are recognized respiratory pathogens. Data accumulated by different laboratories suggest their neurotropic potential. For example, primary cultures of human astrocytes and microglia were shown to be susceptible to an infection by the OC43 strain of HuCV (A. Bonavia, N. Arbour, V. W. Yong, and P. J. Talbot, J. Virol. 71:800-806, 1997). We speculate that the neurotropism of HuCV will lead to persistence within the central nervous system, as was observed for murine coronaviruses. As a first step in the verification of our hypothesis, we have characterized the susceptibility of various human neural cell lines to infection by HuCV-OC43. Viral antigen, infectious virus progeny, and viral RNA were monitored during both acute and persistent infections. The astrocytoma cell lines U-87 MG, U-373 MG, and GL-15, as well as neuroblastoma SK-N-SH, neuroglioma H4, oligodendrocytic MO3.13, and the CHME-5 immortalized fetal microglial cell lines, were all susceptible to an acute infection by HuCV-OC43. Viral antigen and RNA and release of infectious virions were observed during persistent HuCV-OC43 infections ( approximately 130 days of culture) of U-87 MG, U-373 MG, MO3.13, and H4 cell lines. Nucleotide sequences of RNA encoding the putatively hypervariable viral S1 gene fragment obtained after 130 days of culture were compared to that of initial virus input. Point mutations leading to amino acid changes were observed in all persistently infected cell lines. Moreover, an in-frame deletion was also observed in persistently infected H4 cells. Some point mutations were observed in some molecular clones but not all, suggesting evolution of the viral population and the emergence of viral quasispecies during persistent infection of H4, U-87 MG, and MO3.13 cell lines. These results are consistent with the potential persistence of HuCV-OC43 in cells of the human nervous system, accompanied by the production of infectious virions and molecular variation of viral genomic RNA.


title: Conformational states of the severe acute respiratory syndrome coronavirus spike protein ectodomain.

text: The severe acute respiratory syndrome coronavirus enters cells through the activities of a spike protein (S) which has receptor-binding (S1) and membrane fusion (S2) regions. We have characterized four sequential states of a purified recombinant S ectodomain (S-e) comprising S1 and the ectodomain of S2. They are S-e monomers, uncleaved S-e trimers, cleaved S-e trimers, and dissociated S1 monomers and S2 trimer rosettes. Lowered pH induces an irreversible transition from flexible, L-shaped S-e monomers to clove-shaped trimers. Protease cleavage of the trimer occurs at the S1-S2 boundary; an ensuing S1 dissociation leads to a major rearrangement of the trimeric S2 and to formation of rosettes likely to represent clusters of elongated, postfusion trimers of S2 associated through their fusion peptides. The states and transitions of S suggest conformational changes that mediate viral entry into cells.


title: [Risk factors for aerostasis failure in case of lung resection].

text: The results of risk factors analysis of aerostasis failure in 141 patients are presented in the article. All patients were operated in the A.V. Vishnevskogo Institute of Surgery at the period from January 2009 to December 2010. During statistical analysis it was considered surgeries volume, presence of emphysema, respiratory function etc. Also it was defined the dependence of venting duration in patients with residual pleural cavity and long aerostasis failure.


title: COVID-19 IN DIABETES PATIENTS IN UKRAINE: LESSONS FOR DOCTORS AND PATIENTS.

text: World Health Organization recognized coronavirus disease 2019 (COVID-19) as a pandemic on 11 March 2020. The first case of COVID-19 in Ukraine was confirmed on 3 March 2020 in Chernivtsi and the first lethal case was reported on 13 March 2020. As of 22 April 2020 6592 cases of COVID-19 have been reported in Ukraine, among these cases 174 caused death (2.64% lethality), 467 patients (7.08% of cases) have recovered. The feature of COVID-19 pandemics in Ukraine is that the age of patients is lower (50-59 years old, 40-49 years old and 30-39 years old) than in China and other European countries. Another feature of COVID-19 epidemiology in Ukraine is that virus cause disease in women (56.0%) more often than in men (44.0%). But there are more male patients in other countries. Although there are more male deaths (51.7%) than female (48.3%) in Ukraine. Deaths have occurred in 30-49 years old patients (14%), in 50-69 years old patients (48%), in 70 years old and older patients (38%). 85% of all deaths were over 50 years of age. 79% of patients who died from COVID-19 had severe cardiovascular diseases, diabetes mellitus, neoplasms, renal diseases, respiratory disorders and obesity. In total, co-morbidities (one or several) have been cardiovascular diseases (74%), diabetes mellitus (22%), lungs diseases (9%), neoplasms (8%), renal diseases (7%). In this article we analyzed epidemiology and causes of negative influence of diabetes mellitus on COVID-19 progress in world and Ukraine. We also made recommendations on prophylaxis and patients treatment in cases of COVID-19 and diabetes mellitus co-morbidity.


title: Structural genomics of the severe acute respiratory syndrome coronavirus: nuclear magnetic resonance structure of the protein nsP7.

text: Here, we report the three-dimensional structure of severe acute respiratory syndrome coronavirus (SARS-CoV) nsP7, a component of the SARS-CoV replicase polyprotein. The coronavirus replicase carries out regulatory tasks involved in the maintenance, transcription, and replication of the coronavirus genome. nsP7 was found to assume a compact architecture in solution, which is comprised primarily of helical secondary structures. Three helices (alpha2 to alpha4) form a flat up-down-up antiparallel alpha-helix sheet. The N-terminal segment of residues 1 to 22, containing two turns of alpha-helix and one turn of 3(10)-helix, is packed across the surface of alpha2 and alpha3 in the helix sheet, with the alpha-helical region oriented at a 60 degrees angle relative to alpha2 and alpha3. The surface charge distribution is pronouncedly asymmetrical, with the flat surface of the helical sheet showing a large negatively charged region adjacent to a large hydrophobic patch and the opposite side containing a positively charged groove that extends along the helix alpha1. Each of these three areas is thus implicated as a potential site for protein-protein interactions.


title: An observational study exploring amplitude-integrated electroencephalogram and spectral edge frequency during paediatric anaesthesia.

text: Processed electroencephalography is used in adults to guide anaesthesia, but the algorithms used may not apply to infants. Knowledge of infants' electroencephalogram (EEG) responses to anaesthetics is fragmentary. An earlier pilot study suggested amplitude-integrated EEG (aEEG) may be a useful measure of anaesthetic effect. The aim of this study was to determine how aEEG changes between awake and anaesthetised children of varying ages and to compare the response to that seen with Spectral Edge Frequency 90% (SEF90). A prospective observational study of children receiving a general anaesthetic was conducted. Anaesthetic regimen remained at the discretion of the treating anaesthetist. EEG data were collected using the BrainZ ReBrim(TM) monitor using forehead and biparietal montages. SEF90 and aEEG were compared across age groups, EEG montage and between awake and anaesthetised states. A total of 178 children (aged 24 days to 14 years) were recruited. All aEEGs were greater during anaesthesia compared to when awake and this difference varied with age. Only children older than two years showed lower SEF90 while anaesthetised compared to when awake. SEF90 from children younger than six months was higher during anaesthesia compared to when awake. Analysis of parietal and forehead EEG montages revealed age-related differences. These findings suggest that SEF90 and aEEG can discriminate between awake and anaesthetised states in older children. In younger children aEEG changes are less pronounced and SEF90 either cannot discriminate between states or responds paradoxically. The aEEG may be marginally better than other EEG parameters in measuring anaesthetic depth in children.


Infectious diseases. Annual review of significant publications.


title: Influenza among the elderly in the Americas: a consensus statement.

text: Influenza exacts a heavy burden on the elderly, a segment of the population that is estimated to experience rapid growth in the near future. In the past decade most developed and several developing countries have recommended influenza vaccination for those > 65 years of age. The World Health Organization (WHO) set a goal of 75% influenza vaccination coverage among the elderly by 2010, but it was not achieved. In 2011, the Technical Advisory Group at the Pan American Health Organization, Regional Office of WHO for the Americas, reiterated the influenza vaccine recommendation for older adults. Relatively little information has been compiled on the immunological aspect of aging or on reducing its impact, information particularly relevant for clinicians and gerontologist with firsthand experience confronting its effects. To fill this data gap, in 2012 the Americas Health Foundation (Washington, D.C., United States) and the nonprofit, Fighting Infectious Diseases in Emerging Countries (Miami, Florida, United States), convened a panel of Latin American clinicians and gerontologists with expertise in influenza to discuss key issues and develop a consensus statement. The major recommendations were to improve influenza surveillance throughout Latin America so that its impact can be quantified; and to conduct laboratory confirmation of influenza for all patients who have flu-like symptoms and are frail, immunosuppressed, have comorbidities, are respiratory compromised, or have been admitted to a hospital. The panel also noted that: since evidence for antivirals in the elderly is unclear, their use should be handled on a case-by-case basis; despite decreased immunological response, influenza vaccination in older adults is still crucial; indirect immunization strategies should be encouraged; and traditional infection control measures are essential in long-term care facilities.


title: Association of coronavirus infection with neonatal necrotizing enterocolitis.

text: From the clustered occurrence of numerous cases of necrotizing enterocolitis in newborns, it was possible to associate this disease significantly with infection due to coronavirus-like agents. Prematurity or low birth weight did not seem to affect the development of the disease, at least during the present epidemic. However, associated gas-producing bacteria could influence its severity and play a role in the appearance of pneumatosis. In many aspects the human disease is reminiscent of experimental necrotizing enterocolitis obtained by infection of germ-free newborn animals, as reported in the literature.


title: Politics and partnerships: challenges and rewards of partnerships in workplace health research in the healthcare sector of British Columbia, Canada.

text: In British Columbia (BC), Canada, a partnership of researchers, healthcare employers, and healthcare unions reduced high injury rates through examining determinants of healthy workplaces and designing, implementing, and evaluating interventions. Over 51 million dollars (Canadian) was saved from the BC healthcare budget over two years, largely attributable to the collaborative effort. Challenges and rewards of the process were determined from interviews and workshops with researchers and community stakeholders, and by obtaining direct input to this report. Challenges included maintaining communication and trust between partners, preserving partnerships during restructuring and labor disputes, and maintaining involvement and support of front-line workers and senior management. As all partners recognized the importance of the research agenda, the stakeholders remained committed to working through the challenges, and have consequently achieved considerable success.


title: [Studies on the aetiology of neonatal diarrhoea in veal calves (author's transl)].

text: A herd of sixty-two veal calves was studied to determine whether there was a relationship between neonatal diarrhoea and infections with rotavirus, bovine coronavirus, Salmonella and K99-positive E. coli. Examination of the animals was negative for salmonellosis. Clinical findings and serological testing did not suggest that K99-positive E. coli was a factor in aetiology. Bovine coronavirus was isolated from the faeces of a small number of calves, only a few of which were affected with diarrhoea. A few faecal samples were found to harbour bovine coronavirus as well as rotavirus. Rotavirus was isolated from the faeces of a large number of calves; excretion of the virus was only associated with diarrhoea in some cases. A definite conclusion could therefore not be drawn from the findings, although they suggested a possible role of rotavirus in the aetiology of neonatal diarrhoea of veal calves.


title: Role of tight junctions in establishing and maintaining cell polarity.

text: The tight junction (TJ) is not randomly located on the cell membrane, but occupies a precise position at the outermost edge of the intercellular space and, therefore, is itself considered a polarized structure. This article reviews the most common experimental approaches for studying this relationship. We then discuss three main topics. (a) The mechanisms of polarization that operate regardless of the presence of TJs: We explore a variety of polarization mechanisms that operate at stages of the cell cycle in which TJs may be already established. (b) TJs and polarity as partners in highly dynamic processes: Polarity and TJs are steady state situations that may be drastically changed by a variety of signaling events. (c) Polarized distribution of membrane molecules that depend on TJs: This refers to molecules (mainly lipids) whose polarized distribution, although not the direct result of TJs, depends on these structures to maintain such distribution.


title: Acquisition of macrophage tropism during the pathogenesis of feline infectious peritonitis is determined by mutations in the feline coronavirus spike protein.

text: In feline coronavirus (FCoV) pathogenesis, the ability to infect macrophages is an essential virulence factor. Whereas the low-virulence feline enteric coronavirus (FECV) isolates primarily replicate in the epithelial cells of the enteric tract, highly virulent feline infectious peritonitis virus (FIPV) isolates have acquired the ability to replicate efficiently in macrophages, which allows rapid dissemination of the virulent virus throughout the body. FIPV 79-1146 and FECV 79-1683 are two genetically closely related representatives of the two pathotypes. Whereas FECV 79-1683 causes at the most a mild enteritis in young kittens, FIPV 79-1146 almost invariably induces a lethal peritonitis. The virulence phenotypes correlate with the abilities of these viruses to infect and replicate in macrophages, a feature of FIPV 79-1146 but not of FECV 79-1683. To identify the genetic determinants of the FIPV 79-1146 macrophage tropism, we exchanged regions of its genome with the corresponding parts of FECV 79-1683, after which the ability of the FIPV/FECV hybrid viruses to infect macrophages was tested. Thus, we established that the FIPV spike protein is the determinant for efficient macrophage infection. Interestingly, this property mapped to the C-terminal domain of the protein, implying that the difference in infection efficiency between the two viruses is not determined at the level of receptor usage, which we confirmed by showing that infection by both viruses was equally blocked by antibodies directed against the feline aminopeptidase N receptor. The implications of these findings are discussed.


Recovery of transplantable organs after cardiac or circulatory death: the end justifying the means.


title: A retrospective study on poult enteritis syndrome in Minnesota.

text: A retrospective study was conducted to determine the occurrence of poult enteritis syndrome (PES) in Minnesota from January 2002 to December 2007. PES is an infectious intestinal disease of young turkeys between 1 day and 7 wk of age and is characterized by diarrhea, depression, and lethargy with pale intestines and/or excessively fluid cecal contents. During the study period, samples from 1736 turkey flocks were submitted to the Minnesota Veterinary Diagnostic Laboratory for disease investigation. Of these, 151 flocks (8.7%) were PES positive. Cases of PES were seen throughout the year with higher prevalence in fall. The PES was statistically associated with age with higher occurrence in poults less than 3 wk of age. Rotavirus, small round virus (SRV), Salmonella, nonhemolytic Escherichia coli, Enterococcus, and Eimeria oocysts were detected alone or in different combinations. Reovirus and adenovirus were found in one flock each. The most commonly identified pathogens were Salmonella (85 flocks) and rotavirus (73 flocks). Of PES-affected flocks, 39 (25.8%), 66 (43.7%), and 37 (24.5%) had one, two, and three or more pathogens, respectively. Rotavirus, SRV, and reovirus occurred mostly in poults of less than 6 wk of age while Salmonella, E. coli, and Eimeria were seen in poults of all age groups. Minimum age for rotavirus detection was in 2-day-old poults. Histopathologically, moderate to severe mixed intestinal villus or lamina propria inflammatory infiltrates, necrosis of distal villus tips in intestinal specimens, and mild to severe lymphocellular depletion in thymus, bursa, and spleen were seen. Antimicrobial sensitivity patterns of bacterial isolates from PES-affected flocks revealed maximum sensitivity to trimethoprim/sulfamethoxazole and ceftiofur and a varying degree of resistance to other antimicrobials.


title: Instrumentation for minimally invasive surgery in pediatric urology.

text: The success of modern surgery is dependent on the availability of good equipment and instruments. This dependence increases along with the degree of sophistication of the surgery performed. Paediatric minimally invasive and endoscopic surgery are sophisticated techniques where imaging is obtained through a video-circuit. Endoscopic surgery has opened the field of virtual reality in surgery, and in minimally invasive surgery the actual operation is done through a limited number of small holes. Robot-assisted urologic surgery is an emerging and safe technology for many urologic paediatric operations, although further documentation, including long-term functional outcome, is deemed necessary before definite conclusions can be drawn regarding the superiority or not of robotic assistance compared to conventional laparoscopic approaches.


title: Laboratory capability for molecular detection and confirmation of novel coronavirus in Europe, November 2012.

text: A rapid survey by the European Centre for Disease Prevention and Control (ECDC) and the World Health Organization (WHO) Regional Office for Europe ascertained the availability of national reference laboratory testing for a recently detected novel coronavirus as of 28 November 2012. Screening by internal quality controlled upE-RT-PCR assay was available in 23/46 of responding countries in the WHO European Region, of which 19/30 in European Union (EU) and European Economic Area (EEA) countries. Confirmation of positive screened samples by either ORF1b - RT-PCR, or other target RT-PCR assays with sequence analysis or whole-genome sequence analysis was available in 22/46 responding countries of which 18/30 in EU/EEA countries.


title: Forced recombination between distinct strains of Maize streak virus.

text: Recombination between divergent virus genomes is believed to be a major mechanism for generation of novel virus genotypes. We have examined the recombination process in geminiviruses by forcing recombination between two distinct isolates of Maize streak virus (MSV), MSV-Kom and MSV-Set. Heterodimeric agroinfectious constructs containing tandemly cloned mixtures of complete or partial MSV-Set and MSV-Kom genomes were used to simulate a circular dimeric form similar to that which would be expected to occur following a single intermolecular crossing-over event between MSV-Set and MSV-Kom replicative form DNAs at the long intergenic region (LIR)-movement protein gene (MP) interface. We isolated, analysed and biologically characterized many of the recombinant MSV genomes that were generated from the constructs in planta. Apart from having the same simulated breakpoint at the LIR-MP interface, all the genomes examined had a second breakpoint that had been generated through either intramolecular homologous recombination or a replicational release mechanism. The pathogenicities of six predominantly MSV-Kom-like recombinants were tested in maize. While all were capable of producing a symptomatic infection in this host, none was more virulent than MSV-Kom and only two were more virulent than MSV-Set. The two most virulent recombinants were leafhopper transmitted to a range of differentially MSV-resistant maize, wheat and barley genotypes and both were found to have unique biological properties.


title: SARS: lessons learned from other coronaviruses.

text: The identification of a new coronavirus as the etiological agent of severe acute respiratory syndrome (SARS) has evoked much new interest in the molecular biology and pathogenesis of coronaviruses. This review summarizes present knowledge on coronavirus molecular biology and pathogenesis with particular emphasis on mouse hepatitis virus (MHV). MHV, a member of coronavirus group 2, is a natural pathogen of the mouse; MHV infection of the mouse is considered one of the best models for the study of demyelinating disease, such as multiple sclerosis, in humans. As a result of the SARS epidemic, coronaviruses can now be considered as emerging pathogens. Future research on SARS needs to be based on all the knowledge that coronavirologists have generated over more than 30 years of research.


title: Performance analysis of a medical decision algorithm to mitigate spread of SARS due to interfacility patient transfers.

text: OBJECTIVE To determine performance of a medical decision algorithm to mitigate spread of severe acute respiratory syndrome (SARS) from interfacility patient transfers during the Toronto SARS outbreak. METHODS Records from the Provincial Transfer Authorization Centre and Toronto Public Health from April 1 to July 31, 2003, were linked using probabilistic methods. Authorization decision (transfer authorized or denied) and SARS status (probable case, suspect case, or patient under investigation for SARS; or non-SARS case) were obtained for linked records. Primary outcome was the number of patients where correct authorization decisions were made based on SARS status at the time of request. Secondary outcome was the number for whom, in retrospect, authorization decision was correct knowing final SARS status. Algorithm sensitivity, specificity, and predictive values were determined. RESULTS There were 14,571 requests for transfer and 2,132 patients investigated for SARS during the study period. The algorithm authorized 14,551 and did not authorize 20 requests. Sensitivity and specificity to make appropriate authorization decisions at the time of request were 100% (95% confidence interval [CI], 77.2%-100%) and 99.95% (95% CI, 99.9-100%), respectively. Positive and negative predictive values were 65% (95% CI, 44.1%-85.9%) and 100% (95% CI, 98.4%-100%), respectively. Sensitivity and specificity, in retrospect, within ten days of the transfer request were 100% (95% CI, 80.6%-100%) and 99.97% (95% CI, 99.9%-100%), respectively. Positive and negative predictive values were 80% (95% CI, 62.5%-97.5%) and 100% (95% CI, 98.4%-100%), respectively. Seven of the 20 patients with nonauthorized requests were not known to have SARS at the time of request. Within ten days, three of seven were under investigation for, a suspect case of, or a probable case of SARS. CONCLUSIONS The medical decision algorithm was highly sensitive and specific in correctly authorizing transfers. Despite its highly sensitive and specific algorithm, it did incorrectly deny authorization to a very small number of patients without SARS.


title: Needs, gaps and opportunities for infectious disease research in British Columbia: a perspective from population and public health.

text: BACKGROUND A review of infectious disease research activity and capacity was performed in British Columbia and linked to a process for identifying needs, gaps and opportunities from a public health perspective. METHODS The study was organized in three phases: an environmental scan to describe current research activity in BC; a consultation to identify needs, gaps and opportunities with those conducting research (key informants) and the end users of research results (stakeholders); and a prioritization of the research needs emerging from the consultation. RESULTS Analysis and synthesis of the consultation data resulted in the identification of nine research themes, which were prioritized in the following order: efficacy and cost-benefit, disease patterns, emerging infectious disease, immunology and vaccines, disease-specific research, health promotion and communications, safe food and water, knowledge translation research and genomics. Six capacity-building themes were also identified: attraction and retention, education and training, collaboration and networks, funding, dissemination of findings, and public health input, surveillance, informatics and databases. INTERPRETATION The findings were helpful in developing a multi-disciplinary, multi-level infectious disease research agenda linking researchers in universities, hospitals and public health institutions with practitioners and policy-makers in British Columbia's public health system. The approach is both feasible and important to undertake at the national level.


title: Cell-based angiopoietin-1 gene therapy for acute lung injury.

text: RATIONALE The acute respiratory distress syndrome is a significant cause of morbidity and mortality in critically ill patients. Angiopoietin-1 (Ang-1), a ligand for the endothelial Tie2 receptor, is an endothelial survival and vascular stabilization factor that reduces endothelial permeability and inhibits leukocyte-endothelium interactions. OBJECTIVES We hypothesized that Ang-1 counteracts vascular inflammation and pulmonary vascular leak in experimental acute lung injury. METHODS We used cell-based gene therapy in a rat model of ALI. Transgenic mice overexpressing Ang-1 or deficient in the Tie2 receptor were also studied to better elucidate the mechanisms of protection. MEASUREMENTS AND MAIN RESULTS The present report provides data that support a strong protective role for the Ang-1/Tie2 system in two experimental models of LPS-induced acute lung injury. In a rat model, cell-based Ang-1 gene transfer improved morphological, biochemical, and molecular indices of lung injury and inflammation. These findings were confirmed in a gain-of-function conditional, targeted transgenic mouse model, in which Ang-1 reduced endothelial cell activation and the expression of adhesion molecules, associated with a marked improvement in airspace inflammation and intraalveolar septal thickening. Moreover, heterozygous Tie2-deficient mice demonstrated enhanced evidence of lung injury and increased early mortality. CONCLUSIONS These results support a critical role for the Ang-1/Tie2 axis in modulating the pulmonary vascular response to lung injury and suggest that Ang-1 therapy may represent a potential new strategy for the treatment and/or prevention of acute respiratory distress syndrome in critically ill patients.


Success and failure in human virus diseases. I. Aetiology.


title: ADP-ribose-1"-monophosphatase: a conserved coronavirus enzyme that is dispensable for viral replication in tissue culture.

text: Replication of the approximately 30-kb plus-strand RNA genome of coronaviruses and synthesis of an extensive set of subgenome-length RNAs are mediated by the replicase-transcriptase, a membrane-bound protein complex containing several cellular proteins and up to 16 viral nonstructural proteins (nsps) with multiple enzymatic activities, including protease, polymerase, helicase, methyltransferase, and RNase activities. To get further insight into the replicase gene-encoded functions, we characterized the coronavirus X domain, which is part of nsp3 and has been predicted to be an ADP-ribose-1"-monophosphate (Appr-1"-p) processing enzyme. Bacterially expressed forms of human coronavirus 229E (HCoV-229E) and severe acute respiratory syndrome-coronavirus X domains were shown to dephosphorylate Appr-1"-p, a side product of cellular tRNA splicing, to ADP-ribose in a highly specific manner. The enzyme had no detectable activity on several other nucleoside phosphates. Guided by the crystal structure of AF1521, an X domain homolog from Archaeoglobus fulgidus, potential active-site residues of the HCoV-229E X domain were targeted by site-directed mutagenesis. The data suggest that the HCoV-229E replicase polyprotein residues, Asn 1302, Asn 1305, His 1310, Gly 1312, and Gly 1313, are part of the enzyme's active site. Characterization of an Appr-1"-pase-deficient HCoV-229E mutant revealed no significant effects on viral RNA synthesis and virus titer, and no reversion to the wild-type sequence was observed when the mutant virus was passaged in cell culture. The apparent dispensability of the conserved X domain activity in vitro indicates that coronavirus replicase polyproteins have evolved to include nonessential functions. The biological significance of the novel enzymatic activity in vivo remains to be investigated.


title: Intracellular RNAs of the feline infectious peritonitis coronavirus strain 79-1146.

text: In Felis catus whole foetus D cells infected with feline infectious peritonitis virus (FIPV), strain 79-1146, six virus-specific, poly(A)-containing RNA species of about 20, 9.6, 5.2, 3.8, 2.8 and 1.6 kb were found. By translation in vitro the 3.8 and 2.8 kb RNAs were shown to encode the 25K envelope protein and the 45K nucleocapsid protein, respectively. The partial map of the FIPV genome was compared with genomic maps of porcine, murine and avian coronaviruses. Differences in these maps suggest that transcription units have been lost or gained during coronavirus divergence.


title: Kawasaki disease.

text: PURPOSE OF REVIEW Kawasaki disease is still a dilemma for pediatricians, especially its atypical or incomplete presentation. The current diagnostic criteria appear inadequate, and the early diagnosis frequently remains challenging with high risk of coronary damage. This review focuses on recent data concerning Kawasaki disease etiology and pathogenesis, and emphasizes the proposal of including altered laboratory values in revised diagnostic criteria aimed at identifying patients with incomplete disease. RECENT FINDINGS Despite a timely treatment, approximately 15% of children have persistent or recurrent fever, thus requiring additional intravenous immunoglobulin, corticosteroids, immunosuppressant, and eventually biologic drugs. Recent cardiologic tests strongly suggest that endothelial damage may lead to coronary stenosis and myocardial ischemia over time, even in children apparently normal at initial cardiac evaluation. New noninvasive tools are suggested to monitor coronary artery aneurysm outcome. Giant coronary artery aneurysms still represent a severe life-threatening complication. There is as yet no agreement regarding the prevention of thrombosis and stenosis in children with giant aneurysms. SUMMARY Due to earlier recognition, aggressive medical treatment, and surgical procedures, the quality of life in patients with Kawasaki disease is significantly improved. The identification of children at high risk for coronary artery aneurysms is crucial in order to reduce the occurrence of sudden death in adolescence and early adulthood. High parameters of inflammation, anemia, and low sodium and albumin levels, along with persistent unexplained fever, should alert clinicians to suspect the disease even though all clinical symptoms are lacking.


title: The State Laboratory of Hygiene's role in terrorism preparedness and response.

text: In the fall of 2001, the national public health system found itself responding to acts of terrorism. The intentional release of Bacillus anthracis spores on the East Coast tested the capacity of all state public health laboratories to respond. The impact on the public health system extended to the Wisconsin State Laboratory of Hygiene (WSLH). Fortunately, participation in the National Laboratory Response Network helped the WSLH meet the challenge of 24 hour/7 days a week coverage, and subsequent federal funding increases have enabled the WSLH to expand its technical capabilities and provide training and outreach to other Wisconsin laboratories to prepare them for their roles in man-made or naturally-occurring public health emergencies.


title: Pathogenesis of coronavirus-induced infections. Review of pathological and immunological aspects.

text: Coronaviruses and arteriviruses infect multiple species of mammals, including humans, causing diseases that range from encephalitis to enteritis. Several of these viruses infect domestic animals and cause significant morbidity and mortality, leading to major economic losses. In this category are included such pathogens as transmissible gastroenteritis virus, porcine respiratory and reproductive virus and infectious bronchitis virus. The feline coronaviruses (FECV) generally do not cause infections with high morbidity but in a small percentage of cases, the virus mutates to become more virulent. This virus, feline infectious peritonitis virus (FIPV), causes severe disease in young cats. This disease is in large part immunopathological and understanding it is a major goal of coronavirus research.


Dispatches from the front line.


title: Viruses associated with outbreaks of equine respiratory disease in New Zealand.

text: AIM To identify viruses associated with respiratory disease in young horses in New Zealand. METHODS Nasal swabs and blood samples were collected from 45 foals or horses from five separate outbreaks of respiratory disease that occurred in New Zealand in 1996, and from 37 yearlings at the time of the annual yearling sales in January that same year. Virus isolation from nasal swabs and peripheral blood leukocytes (PBL) was undertaken and serum samples were tested for antibodies against equine herpesviruses (EHV-1, EHV-2, EHV-4 and EHV-5), equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3). RESULTS Viruses were isolated from 24/94 (26%) nasal swab samples and from 77/80 (96%) PBL samples collected from both healthy horses and horses showing clinical signs of respiratory disease. All isolates were identified as EHV-2, EHV-4, EHV-5 or untyped EHV. Of the horses and foals tested, 59/82 (72%) were positive for EHV-1 and/or EHV-4 serum neutralising (SN) antibody on at least one sampling occasion, 52/82 (63%) for EHV-1-specific antibody tested by enzyme-linked immunosorbent assay (ELISA), 10/80 (13%) for ERAV SN antibody, 60/80 (75%) for ERBV SN antibody, and 42/80 (53%) for haemagglutination inhibition (HI) antibody to EAdV-1. None of the 64 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. Evidence of infection with all viruses tested was detected in both healthy horses and in horses showing clinical signs of respiratory disease. Recent EHV-2 infection was associated with the development of signs of respiratory disease among yearlings [relative risk (RR)=2.67, 95% CI=1.59-4.47, p=0.017]. CONCLUSIONS Of the equine respiratory viruses detected in horses in New Zealand during this study, EHV-2 was most likely to be associated with respiratory disease. However, factors other than viral infection are probably important in the development of clinical signs of disease.


title: Advantages of minimal incision laparoscopic cholecystectomy.

text: BACKGROUND Modifications to conventional laparoscopic cholecystectomy (CLC) are aimed at decreasing abdominal wall trauma and improving cosmetic outcome. Although single-incision laparoscopic surgery (SILS) provides excellent cosmetic results, the procedure is technically challenging and expensive compared to the conventional laparoscopic approach. OBJECTIVES To describe a novel, hybrid technique combining SILS and conventional laparoscopy using minimal abdominal wall incisions. METHODS Fifty patients diagnosed with symptomatic cholelithiasis were operated using two reusable 5 mm trocars inserted through a single 15 mm umbilical incision and a single 2-3 mm epigastric port. This technique was clubbed "minimal incision laparoscopic cholecystectomy" (MILC). RESULTS MILC was completed in 49 patients (98%). In five patients an additional 3 mm trocar was used and in 2 patients the epigastric trocar was switched to a 5 mmtrocar. The procedure was converted to CLC in one patient. Mean operative time was 29 minutes (range 18-60) and the average postoperative hospital stay was 22 hours (range 6-50). There were no postoperative complications and the cosmetic results were rated excellent by the patients. CONCLUSIONS MILC is an intuitive, easy-to-learn and reproducible technique and requires small changes from CLC. As such, MILC may be an attractive alternative, avoiding the cost and complexity drawbacks associated with SILS.


title: The challenges of "continuing medical education" in a pandemic era.

text: Closure of medical schools or the barring of "live patient" contact during an epidemic or pandemic is potentially disruptive to medical education. During the SARS epidemic, the use of web-based learning, role play, video vignettes and both live and mannequin-based simulated patients minimised disruptions to medical education. This article examines the pedagogical innovations that allow clinical teaching to continue without medical students examining actual patients, and proposes a contingency plan in the event of future outbreaks that may necessitate similar containment measures.


title: Single amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domain.

text: Neutralizing antibodies (NAbs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) spike (S) glycoprotein confer protection to animals experimentally infected with the pathogenic virus. We and others previously demonstrated that a major mechanism for neutralizing SARS-CoV was through blocking the interaction between the S glycoprotein and the cellular receptor angiotensin-converting enzyme 2 (ACE2). In this study, we used in vivo electroporation DNA immunization and a pseudovirus-based assay to functionally evaluate immunogenicity and viral entry. We characterized the neutralization and viral entry determinants within the ACE2-binding domain of the S glycoprotein. The deletion of a positively charged region Sdelta(422-463) abolished the capacity of the S glycoprotein to induce NAbs in mice vaccinated by in vivo DNA electroporation. Moreover, the Sdelta(422-463) pseudovirus was unable to infect HEK293T-ACE2 cells. To determine the specific residues that contribute to related phenotypes, we replaced eight basic amino acids with alanine. We found that a single amino acid substitution (R441A) in the full-length S DNA vaccine failed to induce NAbs and abolished viral entry when pseudoviruses were generated. However, another substitution (R453A) abolished viral entry while retaining the capacity for inducing NAbs. The difference between R441A and R453A suggests that the determinants for immunogenicity and viral entry may not be identical. Our findings provide direct evidence that these basic residues are essential for immunogenicity of the major neutralizing domain and for viral entry. Our data have implications for the rational design of vaccine and antiviral agents as well as for understanding viral tropism.


title: Targeting of active sialyltransferase to the plant Golgi apparatus.

text: Glycosyltransferases in the Golgi apparatus synthesize cell wall polysaccharides and elaborate the complex glycans of glycoproteins. To investigate the targeting of this type of enzyme to plant Golgi compartments, we generated transgenic Arabidopsis plants expressing alpha-2,6-sialyltransferase, a glycosyltransferase of the mammalian trans-Golgi cisternae and the trans-Golgi network. Biochemical analysis as well as immunolight and immunoelectron microscopy of these plants indicate that the protein is targeted specifically to the Golgi apparatus. Moreover, the protein is predominantly localized to the cisternae and membranes of the trans side of the organelle. When supplied with the appropriate substrates, the enzyme has significant alpha-2,6-sialyltransferase activity. These results indicate a conservation of glycosyltransferase targeting mechanisms between plant and mammalian cells and also demonstrate that glycosyltransferases can be subcompartmentalized to specific cisternae of the plant Golgi apparatus.


Colorectal surgery--the Cinderella specialty.


Respiratory tract infection.


title: Global efforts on vaccines for COVID-19: Since, sooner or later, we all will catch the coronavirus.

text: COVID-19 is an emerging infectious disease that has turned into a pandemic. It spreads through droplet transmission of the new coronavirus SARS-CoV-2. It is an RNA virus displaying a spike protein as the major surface protein with significant sequence similarity to SARS-CoV which causes severe acute respiratory syndrome. The receptor binding domain of the spike protein interacts with the human angiotensin converting enzyme 2 and is considered as the antigenic determinant for stimulating an immune response. While multiple candidate vaccines are currently under different stages of development, there are no known therapeutic interventions at the moment. This review describes the key genetic features that are being considered for generating vaccine candidates by employing innovative technologies. It also highlights the global efforts being undertaken to deliver vaccines for COVID-19 through unprecedented international cooperation and future challenges post development.


title: SARS/avian coronaviruses.

text: In the hunt for the aetiology of the SARS outbreak in 2003, a newly developed virus DNA micro-array was successfully used to hybridise PCR products obtained by random amplification of nucleic acids extracted from a cell culture infected with material from a SARS patient. The SARS agent was found to hybridise with micro-array probes from both coronaviruses and astroviruses, but one of the coronavirus probes and the four astrovirus probes contained redundant sequences, spanning a highly conserved motif, named s2m, found at the 3' end of the genomes of almost all astroviruses, one picornavirus, and the poultry coronaviruses. The three other coronavirus probes, that hybridised with the SARS agent, were located in the replicase gene, and it could be concluded that the SARS agent was a novel coronavirus, harbouring s2m. The presence of this motif in different virus families is probably the result of recombinations between unrelated viruses, but its presence in both poultry and SARS coronaviruses could suggest a bird involvement in the history of the SARS coronavirus. A recent screening of wild birds for the presence of coronaviruses, using a pan-coronavirus RT-PCR, led to the identification of novel coronaviruses in the three species studied. Phylogenetic analyses performed on both replicase gene and nucleocapsid protein could not add support to a close relationship between avian and SARS coronaviruses, but all the novel avian coronaviruses were found to harbour s2m. The motif is inserted at a homologous place in avian and SARS coronavirus genomes, but in a somewhat different context for the SARS coronavirus. If the presence of s2m in these viruses is a result of two separate recombination events, this suggests that its particular position in these genomes is the only one that would not be deleterious for coronaviral replication, or that it is the result of a copy-choice recombination between coronaviruses, following an ancestral introduction in the coronavirus family by an unrelated virus. In conclusion, the relative high frequencies of recombination observed both experimentally and in the natural evolution of RNA viruses, indicate that horizontal gene transfer does occur, even between unrelated viruses. This might represent a challenge in the rapid identification of novel pathogens with DNA micro-array techniques.


title: Presentation of severe acute respiratory syndrome (SARS) patients in a screening centre.

text: INTRODUCTION On 22 March 2003, the Ministry of Health, Singapore, designated Tan Tock Seng Hospital as the nationwide severe acute respiratory syndrome (SARS) hospital and its Emergency Department (ED) took over the role as the screening center for SARS on 26 March 2003. We describe the initial clinical characteristics of probable or suspect SARS patients that presented to the ED. METHODS A retrospective study of patients who were admitted through the ED and subsequently diagnosed to have probable SARS and suspect SARS was done. The data of these patients from the ED log were reviewed and analysed. RESULTS From 13 March 2003 to 31 May 2003, 11,461 patients were screened for SARS and 1,386 patients were admitted. Of these, 117 patients were diagnosed to have probable SARS and 146 suspect SARS. Their mean age was 36.7 years (range 1-80). Among these patients, there were 122 men (46.4 percent), and 205 were Singaporeans (77.9 percent). 13 patients had no initial contact history upon presentation to the ED. The mean duration between onset of symptom to presentation to the ED was 3.1 days. Travel history was less common in probable SARS cases than in suspect SARS cases as the epidemic was due mainly to local transmission. Fever was the most common presenting symptom (91.6 percent), and gastrointestinal symptoms were the least (6.9 percent). In the ED, 249 (94.7 percent) patients had chest radiographs and 86 (32.7 percent) had full blood count done. 22.2 percent of probable SARS patients had normal chest radiographs when they first presented to the ED. CONCLUSION The World Health Organisation criteria were important screening tools and admission guides, but should not be strictly followed. It was difficult to differentiate between probable and suspect SARS patients in the ED.


title: Effects of experimental Mycoplasma pulmonis infection on sensory neuropeptides and airway mucosa in the rat.

text: The effect of airway infection on neurogenic inflammation is not known. The present study examines the effect of Mycoplasma pulmonis infection on the sensory neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) in the trigeminal ganglion and in the mucosa of the nose and trachea in rats. We compared germ-free (GF), conventionally raised (CV) and specific pathogen-free (SPF) rats. The concentrations of SP and CGRP in the nasal mucosa were assessed with immunohistochemistry, and their prohormonal transcripts in the trigeminal ganglion were assessed with Northern blot. Mucosa was also processed for light microscopy and electron microscopy. SP-like immunoreactivity was greater in the nasal mucosa of infected animals than in uninfected controls. CGRP-like immunoreactivity was greater in the nasal septum, but not in the nasal turbinate, of infected than uninfected animals. In contrast, no change was evident in the expression levels of the prohormonal transcripts in the trigeminal ganglion. Infected nasal and tracheal mucosa was oedematous and locally infiltrated with inflammatory cells. In the nose of uninfected GF rats, subepithelial lymphoid aggregations were scarce and appeared inactive. We conclude that Mycoplasma pulmonis infection results in increased immunoreactivity of substance P, probably within nerves. There was no clear evidence of increased synthesis of the precursors of substance P and calcitonin gene-related peptide.