title: Efficient and quick inactivation of SARS coronavirus and other microbes exposed to the surfaces of some metal catalysts.

text: OBJECTIVE To study the two metal catalysts Ag/Al2O3 and Cu/Al2O3 that interdict the transmission pathway for SARS and other respiratory infectious diseases. METHODS Two metal catalysts Ag/Al2O3 and Cu/Al2O3 were pressed into wafers. One hundred microL 10(6) TCID50/mL SARS-CoV, 100 microL 10(6) PFU/mL recombinant baculovirus expressing hamster's prion protein (haPrP) protein and roughly 10(6) E. coli were slowly dropped onto the surfaces of the catalyst wafers and exposed for 5 and 20 min, respectively. After eluted from the surfaces of wafers, the infectivity of viruses and propagation of bacteria were measured. The expression of PrP protein was determined by Western blot. The morphological changes of bacteria were observed by electronic microscopy. RESULTS After exposure to the catalysts surfaces for 5 and 20 min, the infectivity of SARS-CoV in Vero cells and baculovirus in Sf9 cells dropped down to a very low and undetectable level, and no colony was detected using bacteria culture method. The expression of haPrP protein reduced to 21.8% in the preparation of Sf9 cells infected with recombinant baculovirus exposed for 5 min and was undetectable exposed for 20 min. Bacterial membranes seemed to be cracked and the cytoplasm seemed to be effluent from cell bodies. CONCLUSION Exposures to the surfaces of Ag/Al2O3 and Cu/Al2O3 destroy the replication and propagation abilities of SARS-CoV, baculovirus and E. coli. Inactivation ability of metal catalysts needs to interact with air, utilizing oxygen molecules in air. Efficiently killing viruses and bacteria on the surfaces of the two metal catalysts has a promising potential for air-disinfection in hospitals, communities, and households.


title: Rapid creation of a temporary isolation ward for patients with severe acute respiratory syndrome in Taiwan.

text: OBJECTIVES To rapidly establish a temporary isolation ward to handle an unexpected sudden outbreak of severe acute respiratory syndrome (SARS) and to evaluate the implementation of exposure control measures by healthcare workers (HCWs) for SARS patients. DESIGN Rapid creation of 60 relatively negative pressure isolation rooms for 196 suspected SARS patients transferred from 19 hospitals and daily temperature recordings of 180 volunteer HCWs from 6 medical centers. SETTING A military hospital. RESULTS Of the 196 patients, 34 (17.3%) met the World Health Organization criteria for probable SARS with positive results of serologic testing for SARS-associated coronavirus (SARS-CoV), reverse transcriptase polymerase chain reaction (RT-PCR) from nasopharyngeal or throat swabs for SARS-CoV, or both. Seventy-four patients had suspected SARS based on unprotected exposure to SARS patients; three of them had positive results on RT-PCR but negative serologic results. The remaining 88 patients did not meet the criteria for a probable or suspected SARS diagnosis. Of the 34 patients with probable SARS, 13 were transferred to medical centers to receive mechanical ventilation due to rapid deterioration of chest x-ray results, and three patients died of SARS despite intensive therapy in medical centers. During the study period, one nurse developed probable SARS due to violation of infection control measures, but there was no evidence of cross-transmission to other HCWs. CONCLUSIONS Despite the use of full personal protection equipment, the facility failed to totally prevent exposures of HCWs to SARS but minimized the risk of nosocomial transmission. Better training and improvements in infection control infrastructure may limit the impact of SARS.


title: Swine hybrid aneurysm model for endovascular surgery training.

text: The aim of this study was to develop a technically simple swine aneurysm-training model by inserting a silicone aneurysm circuit in the cervical vessels. A silicone aneurysm circuit was created by designing multiple aneurysms in size and configuration on a silicone vessel. Five swine underwent surgical implantation of this circuit in the cervical vessels: one end in the common carotid artery and the other in the external jugular vein. Using this model, an aneurysm coiling procedure was simulated under fluoroscopic guidance, roadmapping and digital subtraction angiography. Creating an aneurysm model for training purposes by this method was technically simple and enabled the formation of a wide variety of aneurysms in a single procedure. The quality of the model was uniform and the model was reproducible. Coiling training using this model resembled a realistic clinical situation. The swine hybrid aneurysm-training model was advantageous from the standpoint of technical simplicity in the creation and variety of aneurysms it provided. The swine hybrid aneurysm model may be an additional option for aneurysm coiling training.


title: Avian influenza H5N1 in viverrids: implications for wildlife health and conservation.

text: The Asian countries chronically infected with avian influenza A H5N1 are 'global hotspots' for biodiversity conservation in terms of species diversity, endemism and levels of threat. Since 2003, avian influenza A H5N1 viruses have naturally infected and killed a range of wild bird species, four felid species and a mustelid. Here, we report fatal disseminated H5N1 infection in a globally threatened viverrid, the Owston's civet, in Vietnam, highlighting the risk that avian influenza H5N1 poses to mammalian and avian biodiversity across its expanding geographic range.


title: Cell entry by enveloped viruses: redox considerations for HIV and SARS-coronavirus.

text: For enveloped viruses, genome entry into the target cell involves two major steps: virion binding to the cell-surface receptor and fusion of the virion and cell membranes. Virus-cell membrane fusion is mediated by the virus envelope complex, and its fusogenicity is the result of an active virus-cell interaction process that induces conformation changes within the envelope. For some viruses, such as influenza, exposure to an acidic milieu within the cell during the early steps of infection triggers the necessary structural changes. However, for other pathogens which are not exposed to such environmental stress, activation of fusogenicity can result from precise thiol/disulfide rearrangements mediated by either an endogenous redox autocatalytic isomerase or a cell-associated oxidoreductase. Study of the activation of HIV envelope fusogenicity has revealed new knowledge about how redox changes within a viral envelope trigger fusion. We discuss these findings and their implication for anti-HIV therapy. In addition, to compare and contrast the situation outlined for HIV with an enveloped virus that can fuse with the cell plasma membrane independent of the redox status of its envelope protein, we review parallel data obtained on SARS coronavirus entry.


Monitoring and management of acidosis in calf diarrhoea.


title: Two-year clinical follow-up of the Multicenter Randomized Clinical Trial of Endovascular Treatment for Acute Ischemic Stroke in The Netherlands (MR CLEAN): design and statistical analysis plan of the extended follow-up study.

text: BACKGROUND MR CLEAN was the first randomized trial to demonstrate the short-term clinical effectiveness of endovascular treatment in patients with acute ischemic stroke caused by large vessel occlusion in the anterior circulation. Several other trials confirmed that endovascular treatment improves clinical outcome at three months. However, limited data are available on long-term clinical outcome. We aimed to estimate the effect of endovascular treatment on functional outcome at two-year follow-up in patients with acute ischemic stroke. Secondly, we aimed to assess the effect of endovascular treatment on major vascular events and mortality during two years of follow-up. METHODS MR CLEAN is a multicenter clinical trial with randomized treatment allocation, open-label treatment, and blinded endpoint evaluation. Patients included were 18 years or older with acute ischemic stroke caused by a proven anterior proximal artery occlusion who could be treated within six hours after stroke onset. The intervention contrast was endovascular treatment and usual care versus no endovascular treatment and usual care. The current study extended the follow-up duration from three months to two years. The primary outcome is the score on the modified Rankin scale at two years. Secondary outcomes include all-cause mortality and the occurrence of major vascular events within two years of follow-up. DISCUSSION The results of our study provide information on the long-term clinical effectiveness of endovascular treatment, which may have implications for individual treatment decisions and estimates of cost-effectiveness. TRIAL REGISTRATION NTR1804 . Registered on 7 May 2009; ISRCTN10888758 . Registered on 24 July 2012 (main MR CLEAN trial); NTR5073 . Registered on 26 February 2015 (extended follow-up study).


title: Influenza and corona-virus antibodies in the newborns and their mothers in Mali (1980).

text: In February 1980 blood samples from 91 women and their newborn babies (cord blood) from Mali have been analysed for influenza and corona OC 43 HI antibodies. The titers of influenza HI antibodies were higher in blood samples collected from the mothers at delivery than in their newborns. Considering the GMT values and percentage of persons with protective titers (greater than or equal to 1:40) we may assume that influenza A (H3N2) viruses had been highly active in Mali. Influenza A (H1N1) and B activity may be considered as minor. The prevalence of OC 43 coronaviruses HI antibodies was higher in the cord blood of the newborn babies (95%) than in the blood samples of their mothers (75%).


title: Building resiliency: ensuring business continuity is on the health care agenda.

text: In light of recent disasters and terrorist attacks, private and public organisations alike are becoming increasingly concerned with their ability to continue operating in spite of unforeseen events. This paper describes a project conducted at the Royal Victorian Eye and Ear Hospital to develop a Business Continuity Management (BCM) Framework, and outlines the learning experience. It provides a Framework and describes the key issues to be considered when initiating BCM in a health organisation, concluding that a project management approach can be used to establish a framework for BCM.


title: Animal models for perinatal transmission of HIV-1.

text: Despite progress in the use of antiretroviral drugs, mother-to-child transmission of HIV still remains a serious medical problem in resource-poor areas. There is a need to find the best method for drug delivery to reduce transmission, while keeping the risk of selection for drug-resistant viral variants low. Even when infection is prevented during pregnancy, the risk of acquiring infection by breast feeding remains significant and in some settings, is unavoidable. The ability of antiretroviral drugs or vaccines to limit transmission by breast milk is unknown. HIV vaccines are still in an early phase of development and have not yet been tested in newborns, in part due to concerns about potential of low immunogenicity due to transplacental transfer of maternal antibodies. Alternative strategies have been proposed to limit transmission using passive prophylaxis by human monoclonal antibody, but to insure product safety, trials have been slowed. Due to such concerns, animal models may provide an alternative for testing efficacy in human newborns. In this review, advances made using such models will be compared for mother-to-child transmission of lentivirus with that of HIV-1. In addition, some perspectives on integrating the data obtained from these models as a groundwork for future clinical work will be presented.


title: Kawasaki disease: a rare pediatric pathology in Mexico. Twenty cases report from the Hospital Infantil del Estado de Sonora.

text: BACKGROUND Kawasaki disease (KD) is an etiological illness that is relatively unknown and scarcely identified in Mexico; it affects children mainly aged 1-4 years, evolves with fever, vasculitis in diverse organs, and in the heart the disease mainly affects the coronary arteries. OBJECTIVES Our aim was to inform the clinical findings and evolution of 20 patients diagnosed with KD. MATERIALS AND METHODS We reviewed the patient clinical files retrospectively and descriptively to obtain information with regard to age, sex, clinical signs, laboratory and consultory results, echocardiography findings, complications, evolution during hospitalization, followup, and out-patient ambulatory consultations. RESULTS Eighteen patients were male, two were female, six developed coronary damage, two aortic mitral-valve insufficiency, one pericardial shedding, and one, myocarditis. All patients received gamma globulin treatment with aspirin, and 16 were controlled during 6-8 months after the acute medical profile. CONCLUSIONS The opportune clinical diagnostic it is fundamental to establish an early treatment with gammmaglobuline to avoid injuries in the arterial coronary level. This injury may cause eventualy ischemia or myocardial infarct


Reviews on animal diseases recently published in other journals.


Enhancing influenza surveillance in the Eastern Mediterranean Region: Eastern Mediterranean Acute Respiratory Infection Surveillance (EMARIS) network - third inter-country meeting.


title: Identification of a new membrane-associated polypeptide specified by the coronavirus infectious bronchitis virus.

text: Nucleotide sequences from the third open reading frame of mRNA D (D3) of infectious bronchitis virus (IBV) were expressed in bacteria as part of a fusion protein with beta-galactosidase. Antiserum raised in rabbits against this fusion protein immunoprecipitated from IBV-infected chick kidney or Vero cells a polypeptide of 12.4K, the size expected for a D3-encoded product. The D3 polypeptide is apparently non-glycosylated, and appears to be associated with the membrane fraction of infected cells, as judged by cell fractionation and immunofluorescence.


Leprosy control in China: trends in detection of new cases, 1987-2008.


Mental health impact of severe acute respiratory syndrome: a prospective study.


Caudal Duplication: Management of a Rare Congenital Condition.


title: Host-pathogen interactions: a proteomic view.

text: Host-pathogen interactions reflect the balance of host defenses and pathogen virulence mechanisms. Advances in proteomic technologies now afford opportunities to compare protein content between complex biologic systems ranging from cells to animals and clinical samples. Thus, it is now possible to characterize host-pathogen interactions from a global proteomic view. Most reports to date focus on cataloging protein content of pathogens and identifying virulence-associated proteins or proteomic alterations in host response. A more in-depth understanding of host-pathogen interactions has the potential to improve our mechanistic understanding of pathogenicity and virulence, thereby defining novel therapeutic and vaccine targets. In addition, proteomic characterization of the host response can provide pathogen-specific host biomarkers for rapid pathogen detection and characterization, as well as for early and specific detection of infectious diseases. A review of host-pathogen interactions focusing on proteomic analyses of both pathogen and host will be presented. Relevant genomic studies and host model systems will be also be discussed.


title: Effect of monopolar radiofrequency energy on pacemaker function.

text: BACKGROUND This study aimed to quantify the clinical parameters of mono- and bipolar instruments that inhibit pacemaker function. The specific aims were to quantify pacer inhibition resulting from the monopolar instrument by altering the generator power setting, the generator mode, the distance between the active electrode and the pacemaker, and the location of the dispersive electrode. METHODS A transvenous ventricular lead pacemaker overdrive paced the native heart rate of an anesthetized pig. The primary outcome variable was pacer inhibition quantified as the number of beats dropped by the pacemaker during 5 s of monopolar active electrode activation. RESULTS Lowering the generator power setting from 60 to 30 W decreased the number of dropped paced events (2.3 ± 1.2 vs 1.6 ± 0.8 beats; p = 0.045). At 30 W of power, use of the cut mode decreased the number of dropped paced beats compared with the coagulation mode (0.6 ± 0.5 vs 1.6 ± 0.8; p = 0.015). At 30 W coagulation, firing the active electrode at different distances from the pacemaker generator (3.75, 7.5, 15, and 30 cm) did not change the number of dropped paced beats (p = 0.314, analysis of variance [ANOVA]). The dispersive electrode was placed in four locations (right/left gluteus, right/left shoulder). More paced beats were dropped when the current vector traveled through the pacemaker/leads than when it did not (1.5 ± 1.0 vs 0.2 ± 0.4; p < 0.001). CONCLUSIONS Clinical parameters that reduce the inhibition of a pacemaker by monopolar instruments include lowering the generator power setting, using cut (vs coagulation) mode, and locating the dispersive electrode so the current vector does not traverse the pacemaker generator or leads.


title: [Influenza surveillance in five consecutive seasons during post pandemic period: results from National Influenza Center, Turkey].

text: Influenza surveillance provides data about the characteristics of influenza activity, types, sub-types and antigenic properties of the influenza viruses in circulation in a region. Surveillance also provides for the preparation against potential influenza pandemics with the identification of the genetic properties of viruses and the mutant strains that could pose a threat. In this study, data in the scope of national influenza surveillance carried out by National Influenza Center, Turkey for five consecutive influenza seasons between 2010-2015, following the A(H1N1)pdm09 virus pandemic, have been presented and evaluated. A total of 15.149 respiratory samples, including 8.894 sentinel and 6.255 non-sentinel specimens, during 2010-2015 influenza seasons, within the periods between September and May, were evaluated in our center. All samples were tested using real-time reverse transcriptase PCR (rRT-PCR) for the presence of influenza virus types and subtypes. Within the sentinel influenza surveillance, the samples that were detected negative for influenza viruses, have also been tested for the other respiratory viruses (respiratory syncytial virus, rhinoviruses, paramyxoviruses, coronaviruses) using the same technique. Further analysis, including virus isolation by cell culture inoculation and antigenic characterization by hemagglutination inhibiton test were performed for the samples found positive for influenza A and B viruses. Selected representative virus isolates have been sent to WHO reference laboratory for the sequence analysis. In the study, influenza virus positivity rates detected for all of the samples (sentinel+non-sentinel) were as follows; 34% (779/2316) in 2010-11 season; 25% (388/1554) in 2011-12; 20% (696/3541) in 2012-13; 23% (615/2678) in 2013-14; and 26% (1332/5060) in 2014-15. When all the samples were considered for influenza A and B viruses, the positivity rates for the seasons of 2010-11; 2011-12; 2012-13; 2013-14; 2014-15 were determined as follows; 49.9% and 50.1%, 71.6% and 28.4%; 98.3% and 1.7%; 73.6% and 26.4%; 48.1% and 51.9%, respectively. The frequency of respiratory viruses detected only in sentinel samples other than influenza, were found as follows; 10% (148/1435) in 2010-11; 18% (175/963) in 2011-12; 23% (415/1768) in 2012-13; 22% (468/2108) in 2013-14; and 21% (546/2620) in 2014-15 seasons. When the distribution of influenza virus subtypes were considered, the detection rates of A(H1N1)pdm09 and A(H3N2) viruses in all of the samples were 55% and 45%; 0% and 100%; 95% and 5%; 2% and 98%; and 79% and 21% in the seasons of 2010-11; 2011-12; 2012-13; 2013-14; and 2014-15, respectively. For B/Victoria and B/Yamagata lineages, those rates in the same order of seasons were found as 15% and 85%; 98% and 2%; 0% and 100%; 16% and 84%; and 2% and 98%, respectively. When the data were evaluated, for 2010-2011 and 2014-2015 seasons, cocirculation of influenza A and B were observed within the same periods and similar proportions, but the peak activity occurred 7-8 weeks later for 2014-2015 season. For both seasons, A(H1N1)pdm09 viruses were predominant for non-sentinel, while A(H3N2) viruses were predominant for sentinel detections. During 2011-2012 and 2013-2014 seasons, influenza activity presented similar profile; predominance of A(H3N2) and B viruses observed while A(H1N1)pdm09 virus detections remained low. In contrast, for the 2012-2013 season, A(H1N1)pdm09 viruses were predominant and the detection of A(H3N2) and B viruses remained low. For the seasons in which A(H3N2) was predominant, the peak activity seen earlier than the other seasons. For all seasons, A(H1N1)pdm09 viruses in circulation were antigenically compatible with the vaccine virus, while A(H3N2) viruses were compatible for three seasons (2010-11, 2012-13, 2013-14) and incompatible in two seasons (2011-12, 2014-15). Influenza B viruses were determined as antigenically compatible with the vaccine viruses in all except 2010-2011 season. Predominance of B/Victoria lineage were observed during 2011-2012 season while the rest of the majority were B/Yamagata. The positivity rates of other respiratory viruses which were also analyzed in the scope of sentinel influenza surveillance were similar to influenza positivity for all seasons except 2010-2011. This fact has emphasized that, those viruses were also responsible for influenza-like illness especially during the early and late phases of the season. In conclusion, monitoring of the antigenic and genetic characteristics of influenza viruses by surveillance studies is essential for the early detection of potential pandemic variants as well as to ensure similarities among the circulating strains and the corresponding vaccine strains.


title: Daily Situation Report on Coronavirus disease (COVID-19) in Iran; March 15, 2020.

text: After detection of the first confirmed cases of COVID-19 in Iran, the National Committee on COVID-19 Epidemiology in Ministry of Health and Medical Education was established. This Committee is official source of gathering, analyzing, and reporting the COVID-19 data in Iran. The data of all sources in the country including, medical care monitoring center (MCMC), Hospitals' Information Systems (HIS), Laboratory portal, the data of the center for communicable disease control (MOH), as well as the data from community health centers are integrated and used in this regards. This factsheet contain daily situation report on coronavirus disease (covid-19) in Iran; March 15, 2020.


title: The pathogenesis of ventilator-associated pneumonia: its relevance to developing effective strategies for prevention.

text: Ventilator-associated pneumonia (VAP) is the most common nosocomial infection in the intensive care unit and is associated with major morbidity and attributable mortality. Strategies to prevent VAP are likely to be successful only if based upon a sound understanding of pathogenesis and epidemiology. The major route for acquiring endemic VAP is oropharyngeal colonization by the endogenous flora or by pathogens acquired exogenously from the intensive care unit environment, especially the hands or apparel of health-care workers, contaminated respiratory equipment, hospital water, or air. The stomach represents a potential site of secondary colonization and reservoir of nosocomial Gram-negative bacilli. Endotracheal-tube biofilm formation may play a contributory role in sustaining tracheal colonization and also have an important role in late-onset VAP caused by resistant organisms. Aspiration of microbe-laden oropharyngeal, gastric, or tracheal secretions around the cuffed endotracheal tube into the normally sterile lower respiratory tract results in most cases of endemic VAP. In contrast, epidemic VAP is most often caused by contamination of respiratory therapy equipment, bronchoscopes, medical aerosols, water (eg, Legionella) or air (eg, Aspergillus or the severe acute respiratory syndrome virus). Strategies to eradicate oropharyngeal and/or intestinal microbial colonization, such as with chlorhexidine oral care, prophylactic aerosolization of antimicrobials, selective aerodigestive mucosal antimicrobial decontamination, or the use of sucralfate rather than H(2) antagonists for stress ulcer prophylaxis, and measures to prevent aspiration, such as semirecumbent positioning or continuous subglottic suctioning, have all been shown to reduce the risk of VAP. Measures to prevent epidemic VAP include rigorous disinfection of respiratory equipment and bronchoscopes, and infection-control measures to prevent contamination of medical aerosols. Hospital water should be Legionella-free, and high-risk patients, especially those with prolonged granulocytopenia or organ transplants, should be cared for in hospital units with high-efficiency-particulate-arrestor (HEPA) filtered air. Routine surveillance of VAP, to track endemic VAPs and facilitate early detection of outbreaks, is mandatory.


title: Regionalization in local public health systems: variation in rationale, implementation, and impact on public health preparedness.

text: Comparative case studies found that regionalization originated from a crisis or perceived need for a coordinated response, a need to build local public health capacity, or an effort to use federal preparedness funds more efficiently. Regions vary in terms of their congruence with regional structures for partner agencies, such as emergency management agencies, as well as hospital and health services markets and organizational structure. Some focus on building formal organizational relationships to coordinate and sometimes standardize preparedness and response activities or build regional capacity, while others focus on building informal professional networks. Whatever the approach, strong leadership and trust are required for effective planning, emergency response, and sustainability. This article suggests that regionalization improves emergency preparedness by allowing for more efficient use of resources and better coordination and demonstrated progress in terms of planning and coordination; regional capacity-building, training, and exercises; and development of professional networks.


title: Central role of hypothalamic digoxin in conscious perception, neuroimmunoendocrine integration, and coordination of cellular function: relation to hemispheric dominance.

text: Alteration in the isoprenoid metabolites--digoxin, ubiquinone, and dolichol--have been reported in neuronal degeneration (Parkinson's disease), oncogenesis (central nervous system glioma), functional neuropsychiatric disorders (schizophrenia and epilepsy), and immune-mediated disorders (multiple sclerosis). The coexistence of these disorders has been documented in literature and a central dysfunction related to digoxin and the isoprenoid pathway may underlie all these disorders. A family with a high prevalence of Parkinson's disease, schizophrenia, neoplasms, syndrome X, rheumatoid arthritis, and epilepsy has been described. The psychological behavioral patterns of the family were: creativity and high IQ, hypersexual behavior, reduced appetite and eating behavior, insomnia and reduced sleep patterns, increased tendency for spirituality, increased tendency for addiction, less bonding and affectionate behavior, and left handedness/right hemispheric dominance. Digoxin, an endogenous Na(+)-K+ ATPase inhibitor secreted by the hypothalamus, was found to be elevated and red blood cell (RBC) membrane Na(+)-K+ ATPase activity was found to be reduced in all the disorders and in the indexed family studied. Hypothalamic digoxin can modulate conscious perception and its dysfunction may lead to schizophrenia. Digoxin can also preferentially upregulate tryptophan transport over tyrosine, resulting in increased levels of depolarizng tryptophan catabolites, serotonin, quinolinic acid, strychnine, and nicotine, and decreased levels of hyperpolarizing tyrosine catabolites, dopamine, noradrenaline, and morphine, contributing to membrane Na(+)-K+ ATPase inhibition in all the above disorders and the indexed family. Digoxin-induced membrane Na(+)-K+ ATPase inhibition can result in increased intracellular Ca2+ and reduced Mg2+ levels, leading on to glutamate excitotoxicity, oncogene activation, and immune activation. Digoxin-induced altered Ca2+/Mg2+ ratios, reduced ubiquinone, and increased dolichol can affect glycoconjugate metabolism, membrane formation and structure, and mitochondrial function, leading to the diverse disorders described above, including those in the indexed family. The isoprenoid pathway and neurotransmitter patterns were compared in right-handed/LH dominant and left-handed/RH dominant individuals. The left-handed/RH dominant individuals compared to right-handed/LH dominant individuals had elevated hydroxymethylglutarylcoenzyme A reductase activity, with increased serum digoxin and dolichol levels. The serum ubiquinone, serum Mg2+ and RBC Na(+)-K+ ATPase activity were reduced in left-handed/RH dominant individuals. The left-handed/RH dominant individuals compared to right-handed/LH dominant individuals had elevated levels of serum tryptophan, quinolinic acid, serotonin, nicotine, and strychnine. The levels of tyrosine, dopamine, noradrenaline, and morphine were low in left-handed/RH dominant compared to right-handed/LH dominant individuals. The hyperdigoxinemic state indicates right hemispheric dominance. Hypothalamic digoxin can thus function as the master conductor of the neuroimmunoendocrine orchestra and coordinate the functions of various cellular organelles.


title: Bronchiolitis obliterans organizing pneumonia in a patient with non-Hodgkin's lymphoma following R-CHOP and pegylated filgrastim.

text: Bronchiolitis obliterans organizing pneumonia (BOOP) presents with fever, dyspnoea, pleuritic chest pain and hypoxia. The diagnosis can be made from radiological appearances on chest radiograph and CT scan correlated with histological findings following biopsy. We present a 52-year-old gentleman undergoing treatment for high grade non-Hodgkin's lymphoma who developed respiratory symptoms during chemotherapy. BOOP was diagnosed and he responded well to oral prednisolone. The cause of BOOP is often not certain. However, in this case we suspect pegylated filgrastim or rituximab as possible agents.


title: Clinical and Epidemiological Features of SARS-CoV-2 Patients in SARI Ward of a Tertiary Care Centre in New Delhi.

text: Importance Rapid spread of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in Wuhan, China, prompted heightened surveillance in India. Since the first laboratory confirmed case of SARS-CoV-2 was reported from Kerala on January 30, 2020 novel coronavirus infected pneumonia (NCIP) has been presenting to the hospital emergencies as severe acute respiratory illness (SARI). We aim to find out the rate of SARS-CoV-2 positivity in SARI cases and further clarify the epidemiological and clinical characteristics of NCIP in New Delhi, India. Aims and Objectives To find out the rate of SARS-CoV-2 positivity in SARI cases presenting to the hospital emergency and describe the epidemiological and clinical characteristics of NCIP. Design, Setting and Participants Retrospective, single-center case series of the 82 consecutive hospitalized patients with SARI and subsequent confirmed NCIP cases at Dr Ram Manohar Lohia Hospital, New Delhi between 10th April 2020 and 30th April 2020. Main Outcomes and Measures Epidemiological, demographic, clinical, laboratory, radiological, and treatment data were collected and analyzed. The primary composite end-point was admission to an intensive care unit (ICU), the use of mechanical ventilation or death. Patients were categorized as severe pneumonia and non-severe pneumonia at time of admission and outcome data was compared. Results Of the 82 SARI cases, 32(39%) patients were confirmed to be SARS-CoV-2 positive. The median age of NCIP cases was 54.5 years (IQR, 46.25 - 60) and 19(59.3%) of them were males. 24(75%) cases were categorized as severe pneumonia on admission. 22(68.8%) patients had 1 or more co-morbidities. Diabetes mellitus 16(50%), hypertension 11(34.4%) and chronic obstructive airway disease 5(15.6%) were the most common co-existing illnesses. Compared with the patients who did not meet the primary outcome, patients who met the primary outcome were more likely to be having at least 1 underlying comorbidity (p-0.03), diabetes (p-0.003) and hypertension (p-0.03). Common symptoms included dyspnea 29(90.6%) followed by cough 27(84.4%), fever 22(68%), bodyache and myalgias 14(43.75%). Median time from symptom onset to hospital admission was 3 days. The most common pattern on chest X-ray was bilateral patchy nodular or interstitial infiltration seen in 30(93.8%) patients. Leucopenia was present in 10(31.2%) of the patients, with majority of patients presenting with lymphocytopenia, 24(75%) [lymphocyte count (1106 cells/ dL), interquartile range {IQR}, (970-1487)]. Thrombocytopenia was seen in 14(43.8%) patients, pancytopenia in 10(31.2%) patients and anemia was seen in 14(43.8%) patients. Hypoalbuminemia was present in 22(68.8%) cases. Raised CK-MB was seen in 7(21.9%) patients. The primary composite end-point occurred in 12(37.5%) patients, including 9(28.13%) patients who required mechanical ventilation and subsequently expired. 3(9.3%) of these patients who recovered, were subsequently shifted to COVID-19 ward from the ICU. The patients who met the primary outcome were older in age (56.5 years vs 50 years), had significantly higher SOFA scores (6 vs 3.5), were in shock (41.7% vs 5%), in higher respiratory distress (66.7% vs 10%), had lower mean arterial oxygen saturation (85% vs 89.5%), had higher CK-MB values (66 vs 26)U/L [6(54.5%) vs 2(9.5%)], had hypoalbuminemia (100% vs 50%) and acute kidney injury 8(72.7%) vs 5(23.8%) on admission. Of the 50 non-COVID-19 SARI patients in our study cohort, 13 (26%) patients met the primary composite outcome. Of them 9 (18%) patients expired and remaining 4 patients have subsequently recovered. As on 17th May 2020, 23 patients were still hospitalized, recovering in COVID-19 ward. Conclusion and Relevance In this single-center case series from New Delhi, out of 82 patients of SARI, 32 patients were confirmed NCIP, with a COVID-19 positivity of 39%. 75% of NCIP presented in severe pneumonia and 37.5% required ICU care. The case fatality rate was 28%.


title: [The properties of simian coronavirus].

text: Properties of 20 coronavirus (CV) isolates obtained from spontaneously infected Papio hamadryas and rhesus monkeys were investigated. Two of them were selected as simian CV prototype strains-CVRM 281 (rhesus monkey) and CVP 250 (Papio hamadryas), and can be offered as candidates to the Coronaviridae family. They are closely related to each other, but differ in some biological properties and polypeptides. These strains belong to the 2nd antigen group of mammalian CV with the prototype strain HCV OC 43 but differ from the latter. The strain CVRM 281 induces experimental CV infection which can be used as a model for investigations of some obscure aspects of human infection. The properties of these viruses suggest their usefulness for diagnostic purposes.


Cloning and sequencing the nucleocapsid and E1 genes of coronavirus.


title: Air conditioning systems as non-infectious health hazards inducing acute respiratory symptoms.

text: Chronic and acute exposure to toxic aerosols belongs to frequent causes of airway diseases. However, asthma attacks due to long-distance inhalative exposure to organic solvents, transmitted via an air condition system, have not been reported so far. The present case illustrates the possibility of air conditioning systems as non-infectious health hazards in occupational medicine. So far, only infectious diseases such as legionella pneumophila pneumonia have commonly been associated to air-conditioning exposures but physicians should be alert to the potential of transmission of toxic volatile substances via air conditioning systems. In view of the events of the 11th of September 2001 with a growing danger of large building terrorism which may even use air conditioning systems to transmit toxins, facility management security staff should be alerted to possible non-infectious toxic health hazards arising from air-conditioning systems.


title: Production of high-affinity human monoclonal antibody fab fragments to the 19-kilodalton C-terminal merozoite surface protein 1 of Plasmodium falciparum.

text: A combinatorial immunoglobulin gene library was constructed from peripheral blood lymphocytes of eight patients infected with Plasmodium falciparum and was screened for the production of human monoclonal antibody Fab fragments to the C-terminal 19-kDa fragment of P. falciparum merozoite surface protein 1 (MSP-1(19)). Three Fab clones recognized recombinant MSP-1(19) under nonreducing conditions. Indirect immunofluorescence microscopy demonstrated that three Fab clones stained the surfaces of late trophozoites/schizonts and merozoites of the FCR3 and 3D7 strains, suggesting the Fabs' reactivities to a conserved epitope. Sequence analysis of the heavy-chain genes revealed that the closest germ line V segments were VH1-8 and VH7-81, with 91% to 98% homology. The closest germ line D segment was D3-10, and the closest germ line J segment was JH4 or JH5, with 90% to 97% homology. In the light-chain genes, the closest germ line V segment was A27 for the Jkappa2, Jkappa4, and Jkappa5 segments. The dissociation constants of these Fab fragments for recombinant MSP-1(19) ranged from 1.09 x 10(-9) to 2.66 x 10(-9) M. The binding of the three Fab fragments to MSP-1(19) was competitively inhibited by the anti-MSP-1(19) mouse monoclonal antibody 12.8, which inhibits erythrocyte invasion by merozoites. However, the human Fab fragment with the highest affinity did not inhibit in vitro growth of P. falciparum. This is the first report of gene analysis and bacterial expression of human monoclonal antibodies to P. falciparum MSP-1(19). The combinatorial immunoglobulin gene library derived from malaria patients provides a potential tool for producing high-affinity human antibodies specific for P. falciparum.


title: Global Collaboration in Times of COVID-19: Cuba's Emergency Medical Contingent.

text: The days are long and arduous, with end-less patients to attend, often in a foreign language, always on foreign shores. Far from family and the familiar. Sleep is fi tful at best for health profession-als serving in emergency situations-when sickness obeys no clock and patients' pain haunts even the quiet moments. The crisis scenario varies: post-earthquake, hurricane or tsunami; amid a cholera or Ebola epidemic. The countries vary: Haiti, Pakistan, Guatemala, Mozambique, Sierra Leone. What does not vary is the answer to the calls for help and Cuban professionals' commitment to care for the most vulner-able. These aren't armchair musings or a political pat on the back: they are my own conclusions after living for weeks in close-quarter tents with Cuban doctors, nurses and biomedical engineers in post-earth-quake Pakistan and Haiti, and witnessing their work.


Delayed care-seeking for non-COVID illnesses in Rhode Island.


title: Expression and pharmacological characterization of a stimulatory subtype of adenosine receptor in fetal chick ventricular myocytes.

text: Ventricular and atrial myocytes cultured from chick embryos 14 days in ovo were used as model systems to study cardiac adenosine receptors. In membranes of ventricular cultures, blocking of the A1-adenosine receptor pathway by the A1-selective antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) or by pertussis toxin treatment of the myocyte resulted in a significant adenosine agonist-mediated stimulation of the adenylate cyclase activity. The maximal increases in adenylate cyclase activity caused by the equipotent or the A2-adenosine receptor-selective agonists (from 52.1 +/- 3% to 63 +/- 10% [mean +/- SEM]) were significantly greater than those caused by the A1-selective agonists (from 11 +/- 5% to 34.6 +/- 7%) (p less than 0.01, by t test, n = 4-8). However, in membranes of atrial myocytes, when A1-subtype had been blocked, the various adenosine agonists had no effect on the adenylate cyclase activity. Whether the stimulatory adenylate cyclase-coupled adenosine receptor is also capable of stimulating contractility in the intact ventricular myocyte was next investigated. In ventricular but not in atrial cells, the various adenosine agonists caused an increase in the contractile amplitude in the presence of DPCPX or in myocytes preexposed to pertussis toxin. The increase in contraction amplitude caused by each agonist was expressed as percent of maximum (maximum is the increase in contractility caused by 2.4 mM calcium). In the pertussis toxin-treated myocyte, the maximal increases caused by the equipotent or A2-agonists (NECA, MECA, CV-1808, and CGS21680, from 49.6 +/- 3% to 52.5 +/- 6%, n = 8-12) were significantly greater than those elicited by the A1-agonists (2-CADO, S-PIA, R-PIA, and DCCA, from 12 +/- 4% to 37 +/- 3%, n = 8) (p less than 0.05, by t test). These data demonstrated that a stimulatory adenosine receptor, likely the A2-adenosine receptor, was present on the ventricular but not the atrial myocytes and was linked directly to a stimulation of the cardiac contractility. The functional effects mediated by the A1-subtype became manifested in the presence of isoproterenol, as evidence by an inhibition of the isoproterenol-stimulated increases in adenylate cyclase activity and in cardiac contractility by adenosine agonists. Thus, both subtypes of adenosine receptors, each mediating opposing responses, were present on the ventricular myocytes, whereas only the A1-subtype was found in the atria. The presence of a stimulatory functional A2-adenosine receptor may help explain the absence of a direct negative inotropic response to adenosine in the ventricle.


title: A prospective comparison of the epidemiological and clinical characteristics of pandemic (H1N1) 2009 influenza A virus and seasonal influenza A viruses in Guangzhou, South China in 2009.

text: Comparisons of the clinical characteristics of contemporaneous pandemic (H1N1) 2009 influenza A virus (A(H1N1)pdm09)- and seasonal influenza viruses-infected patients are important for both clinical management and epidemiological studies. A prospective multicenter observational study was conducted using a preestablished sentinel surveillance system in Guangzhou, China during 2009. In this study, the clinical presentations of patients with either acute respiratory infection or community-acquired pneumonia were recorded, and nasopharyngeal swab samples were collected for detection of respiratory virus strains using cell cultures or real-time reverse transcription/real-time polymerase chain reaction. Comparisons of the clinical features between A(H1N1)pdm09- and seasonal influenza viruses-infected patients were conducted accordingly. Of the 1,498 patients examined, 265 tested positive for A(H1N1)pdm09, 286 were positive for seasonal influenza A viruses, and 137 for influenza B viruses. The predominant virus was influenza B before the emergence of A(H1N1)pdm09 (epidemiological week [EW] 1-EW 21); then, predominantly non-A(H1N1)pdm09 influenza A and, later, A(H1N1)pdm09, which peaked in EW 46. Compared with the common seasonal influenza-infected patients, A(H1N1)pdm09-infected patients were younger, and had a higher proportion of these patients reported prior contact with infected individuals (P < 0.001, by χ(2) test). However, few significant differences were observed in clinical symptoms and severity among any of the infections caused by the different influenza A strains. Our hospital-based network served as a useful source of information during A(H1N1)pdm09 monitoring. Viral distribution in Guangzhou was characterized by a sharp rise in A(H1N1)pdm09-infected patients in September 2009. Similar to seasonal influenza A-infected cases, A(H1N1)pdm09 cases had a very small proportion of severe cases.


title: Blessings in disguise: public health emergency preparedness for World Youth Day 2008.

text: Mass gatherings, such as World Youth Day (WYD) 2008 in Sydney, provide an opportunity for public health workers to gain emergency management experience. Communicable disease (mainly influenza) among pilgrims was the major public health issue during WYD. Public health workers also identified environmental health issues, resulting in hazard reduction measures being made to reduce falls. Other public health issues highlighted include training, communication, surge capacity, and health education. Lessons learned from planning WYD mass accommodation could be applied to emergency accommodation in the case of evacuation.


title: [Development and comparison of real-time and conventional RT-PCR assay for detection of human coronavirus NL63 and HKU1].

text: We designed specific primers and fluorescence-labeled probes to develop real-time and conventional RT-PCR assays for detection of human coronavirus NL63 or HKU1. Subsequently, experiments were undertaken to assess diagnostic criteria such as specificity, sensitivity and reproducibility. The detection limit of the real-time RT-PCR assays was 10 RNA copies per reaction mixture. No cross-reactivity was observed between RNA samples derived from designed HCoV and other HCoV or human metapneumovirus. A total of 158 nasopharyngeal swab specimens collected from adult patients with acute respiratory tract infection in Beijing were screened for the presence of human coronavirus NL63 and HKU1 by using real-time RT-PCR and conventional RT-PCR method. The fluorescence quantitative RT-PCR method detected six specimens positive for human coronavirus NL63, five specimens positive for human coronavirus HKU1; and conventional RT-PCR method detected three HCoV-NL63 positive and three HCoV-HKU1 positive, respectively. The convention RT-PCR products of positive samples were obtained and sequence analysis confirmed the reliability of the above methods. In summary, the real-time RT-PCR assay for HCoV- NL63 or HKU1 was more sensitive than conventional RT-PCR and with less time (less than 4 hours) for completion. It may be suitable for molecular epidemiological surveillance and clinical diagnosis for human coronavirus NL63 and HKU1.


title: Analysis of ACE2 in polarized epithelial cells: surface expression and function as receptor for severe acute respiratory syndrome-associated coronavirus.

text: The primary target of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is epithelial cells in the respiratory and intestinal tract. The cellular receptor for SARS-CoV, angiotensin-converting enzyme 2 (ACE2), has been shown to be localized on the apical plasma membrane of polarized respiratory epithelial cells and to mediate infection from the apical side of these cells. Here, these results were confirmed and extended by including a colon carcinoma cell line (Caco-2), a lung carcinoma cell line (Calu-3) and Vero E6 cells in our analysis. All three cell types expressed human ACE2 on the apical membrane domain and were infected via this route, as determined with vesicular stomatitis virus pseudotypes containing the S protein of SARS-CoV. In a histological analysis of the respiratory tract, ACE2 was detected in the trachea, main bronchus and alveoli, and occasionally also in the small bronchi. These data will help us to understand the pathogenesis of SARS-CoV infection.


title: Genetic resistance to mouse hepatitis virus correlates with absence of virus-binding activity on target tissues.

text: The molecular mechanism of genetic resistance of inbred mouse strains to mouse hepatitis virus, a murine coronavirus, was studied by comparing virus binding to plasma membranes of intestinal epithelium or liver from susceptible BALB/c and resistant SJL/J mice with a new solid-phase assay for virus-binding activity. Virus bound to isolated membranes from susceptible mice, but not to membranes from resistant mice. F1 progeny of SJL/J X BALB/c mice had an intermediate level of virus-binding activity on their enterocyte and hepatocyte membranes. This correlated well with previous studies showing that susceptibility to mouse hepatitis virus strain A59 is controlled by a single autosomal dominant gene (M. S. Smith, R. E. Click, and P. G. W. Plagemann, J. Immunol. 133:428-432). Because virus binding was not prevented by treating membranes with sodium dodecyl sulfate, the virus-binding molecule could be identified by a virus overlay protein blot assay. Virus bound to a single broad band of Mr 100,000 to 110,000 in membranes from hepatocytes or enterocytes of susceptible BALB/c and semisusceptible C3H mice, but no virus-binding band was detected in comparable preparations of resistant SJL/J mouse membranes. Therefore, SJL/J mice may be resistant to mouse hepatitis virus A59 infection because they lack a specific virus receptor which is present on the plasma membranes of target cells from genetically susceptible BALB/c and semisusceptible C3H mice.


title: Stat-3 is required for pulmonary homeostasis during hyperoxia.

text: Acute lung injury syndromes remain common causes of morbidity and mortality in adults and children. Cellular and physiologic mechanisms maintaining pulmonary homeostasis during lung injury remain poorly understood. In the present study, the Stat-3 gene was selectively deleted in respiratory epithelial cells by conditional expression of Cre-recombinase under control of the surfactant protein C gene promoter. Cell-selective deletion of Stat-3 in respiratory epithelial cells did not alter prenatal lung morphogenesis or postnatal lung function. However, exposure of adult Stat-3-deleted mice to 95% oxygen caused a more rapidly progressive lung injury associated with alveolar capillary leak and acute respiratory distress. Epithelial cell injury and inflammatory responses were increased in the Stat-3-deleted mice. Surfactant proteins and lipids were decreased or absent in alveolar lavage material. Intratracheal treatment with exogenous surfactant protein B improved survival and lung histology in Stat-3-deleted mice during hyperoxia. Expression of Stat-3 in respiratory epithelial cells is not required for lung formation, but plays a critical role in maintenance of surfactant homeostasis and lung function during oxygen injury.


title: [Preparation and characterization of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein].

text: OBJECTIVE To obtain monoclonal antibodies (McAbs) against severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV) nucleocapsid (N) protein to develop diagnostic test for SARS and study the pathogenesis of the disease. METHODS BALB/c mice were immunized with purified N protein of SARS-CoV. Hybridoma cell lines secreting monoclonal antibodies against SARS-associated coronavirus nucleocapsid were established after cell fusion with mouse splenic cells and SP2/0 cells. The specificity of the McAbs obtained was examined by Western blot and indirect fluorescence assay. Epitopes reacted with the McAbs were preliminarily located through Western blot by expressing truncated N proteins. RESULTS After cell fusion and three rounds of cell cloning, six hybridoma cell lines secreting monoclonal antibodies specifically against SARS-CoV nucleocapsid were obtained. Western blot and indirect fluorescence assay showed that the McAbs reacted specifically with nucleocapsid protein and SARS-CoV. Among the six McAbs, three recognize the epitopes located in the N-terminus of the protein, whereas the others reacted with those located in the C-terminus. CONCLUSION The anti-SARS-CoV nucleocapsid McAbs were developed and these McAbs may be useful in the development of diagnosis assays and basic research of SARS.


title: Serosurvey of small carnivores in the Bolivian Chaco.

text: Five species of Bolivian carnivores, including nine Geoffroy's cats (Oncifelis geoffroyi), ten ocelots (Leopardus pardalis), one jaguarundi (Herpailurus yaguarondi), nine pampas foxes (Pseudalopex gymnocercus), and five crab-eating foxes (Cerdocyon thous) were sampled between March 2001 and April 2005 and tested for antibodies to common pathogens of domestic carnivores. Carnivores were trapped in three areas: a village, the region between human settlements and a protected area, and within Kaa-Iya National Park, Bolivia. Antibodies to canine distemper virus were detected in ocelots and pampas foxes. Antibodies to canine parvovirus were detected in pampas foxes and crab-eating foxes. Geoffroy's cats and all of the ocelots tested positive for antibodies to feline calicivirus (FCV), while fewer than half of Geoffroy's cats and no ocelots had antibodies to feline panleukopenia (FPV). These results confirm that these species of Bolivian carnivores are not naïve to common pathogens of domestic carnivores, and seropositive animals were found in villages as well as in the national park.


title: Detection and characterization of bovine coronaviruses in fecal specimens of adult cattle with diarrhea during the warmer seasons.

text: Bovine coronavirus (BCoV) is an etiological agent associated with winter dysentery (WD), prevalent in adult cattle during the winter. Although we previously detected, isolated, and characterized BCoV strains from adult cattle with WD (WD-BCoV strains) during the winter in South Korea, the precise epidemiology, as well as the causative agent of diarrhea in adult cattle in the warmer seasons, has not been examined. We examined 184 diarrheic fecal specimens collected from 75 herds of adult cattle from seven provinces during the spring (warm), autumn (warm), and summer (hot) seasons. Bovine coronavirus-positive reactions were detected for 107 (58.2%) diarrheic fecal samples (in 47/75 herds). Of these 107 positive samples, 90 fecal samples from 33 herds tested positive for BCoV alone and 17 fecal samples from 14 herds also tested positive for other pathogens. Biological comparisons between the 9 BCoV strains isolated in this study and the 10 previously isolated WD-BCoV strains revealed that there was no receptor-destroying enzyme (RDE) activity against mouse erythrocytes in the 9 BCoV strains but the 10 WD-BCoV strains had high RDE activity. Phylogenetic analysis of the spike (S) and hemagglutinin/esterase (HE) proteins revealed that all the Korean BCoVs clustered together regardless of season and were distinct from the other known BCoVs, suggesting a distinct evolutionary pathway for the Korean BCoVs. These and previous results revealed a high prevalence and widespread geographical distribution of BCoV, suggesting that this virus is endemic in adult cattle with diarrhea in all seasons in South Korea.


SARS: understanding the coronavirus: apoptosis may explain lymphopenia of SARS.


Committee to Advise on Tropical Medicine and Travel (CATMAT). Tuberculosis screening and the international traveller.


title: First isolation of cytopathogenic bovine torovirus in cell culture from a calf with diarrhea.

text: A cytopathogenic virus (designated the Aichi/2004 strain) was isolated in a human rectal adenocarcinoma cell line (HRT-18) from the ileum contents of a calf with diarrhea. Oval and elongated particles, approximately 100 to 170 nm in diameter, with club-shaped projections were seen in the infected culture supernatant, and torovirus-like (tubular and torus nucleocapsid) structures were seen in the infected cells by electron microscopy. An antiserum against bovine torovirus (BToV) reacted with the infected cells by immunofluorescence and neutralized the isolate. However, antisera against bovine coronavirus (BCV) failed to react with the infected cells by immunofluorescence or did not neutralize the isolate. Further, the isolate was positive for BToV by reverse transcription-PCR (RT-PCR) targeting fragments of the nucleocapsid (N), membrane (M), and spike (S) genes. Comparison of the nucleotide sequences of the PCR products with those of the published N, M, and S genes (476 to 497, 672, and 687 to 690 nucleotides, respectively) of toroviruses showed high sequence identities (up to 99.4%, 98.7%, and 94.9% for the N, M, and S genes, respectively) between the isolate and BToVs. In contrast, the isolate was negative for BCV by RT-PCR. In a serological survey of serum samples from 355 calves at 33 farms, 92% of calves were positive for neutralizing antibodies to the isolate. These results indicate that the isolate in this study was BToV and that BToV infection might be common in cattle in Japan. To our knowledge, this is the first isolation of BToV in tissue culture.


title: Coronavirus polyprotein processing.

text: MHV gene 1 contains two ORFs in different reading frames. Translation proceeds through ORF 1a into ORF 1b via a translational frame-shift. ORF 1a potentially encodes three protease activities, two papain-like activities and one poliovirus 3C-like activity. Of the three predicted activities, only the more amino terminal papain-like domain has been demonstrated to have protease activity. ORF 1a polypeptides have been detected in infected cells by the use of antibodies. The order of polypeptides encoded from the 5' end of the ORF is p28, p65, p290. p290 is processed into p240 and p50. Processing of ORF1a polypeptides differs during cell free translation of genome RNA and in infected cells, suggesting that different proteases may be active under different conditions. Two RNA negative mutants of MHV-A59 express greatly reduced amounts of p28 and p65 at the non-permissive temperature. These mutants may have defects in one or more viral protease activities. ORF 1b, highly conserved between MHV and IBV, potentially contains polymerase, helicase and zinc finger domains. None of these activities have yet been demonstrated. ORF 1b polypeptides have yet been detected in infected cells.


London calling?


MHV-A59 pathogenesis in mice.


Data-free modeling of HIV transmission in Sub-Saharan Africa.


title: [Antibody response of patients with severe acute respiratory syndrome (SARS) to nucleocapsid antigen of SARS-associated coronavirus].

text: OBJECTIVE To assess serum antibody responses of patients with severe acute respiratory syndrome (SARS) to nucleocapsid (N) antigen of SARS-associated coronavirus. METHODS The serum levels of IgM and IgG antibodies to N antigen were measured in 200 healthy blood donors and 13 SARS patients at different time points of acute and convalescent phases using indirect enzyme-linked immunosorbent assay (ELISA) with N fusion proteins of SARS-associated coronaviruses. RESULTS The IgM positive critical value of 0.233 and IgG of 0.239 were selected as the threshold value for positive results that equals the product of 2.1 and the mean IgM and IgG levels of 200 healthy blood donors. In 13 patients with SARS, the antibody responses to N antigen were not detectable in the first week after the onset of symptoms. The IgM and IgG seroprotection rates were 83.3% and 66.7% respectively in the second week, both reaching 100% at the third week. IgM seroprotection rates was 61.5% in the second month, and 38.5% at third month. The IgG peaked one month after the onset and remained at high levels in the following 2 months. CONCLUSION The antibody responses suggest that N protein of SARS is immunodominant and plays an important role in viral pathogenesis. This ELISA-based test for detecting anti- N antigen may be of significant value for SARS diagnosis.


title: Comparison of a monoclonal antibody capture ELISA (MACELISA) to indirect ELISA and virus neutralization test for the serodiagnosis of transmissible gastroenteritis virus.

text: An enzyme-linked immunosorbent assay (ELISA) in which the antigen is captured to the plate by monoclonal antibodies (MACELISA) was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). The viral antigen was semipurified from TGEV-infected cells by simple ultracentrifugation. MACELISA results with 258 field sera were compared with those of a standard indirect ELISA and with the virus neutralization test (VNT). Sensitivity, specificity, and kappa values of MACELISA indicated a strong correlation with VNT results, whereas an indirect ELISA was less sensitive and much less specific than VNT. The serologic response of 4 pigs orally inoculated and intraperitoneally boostered with TGEV was compared using the 3 tests. Its sensitivity, specificity, and ability to use unpurified antigen make the MACELISA the advisable first step in TGEV serodiagnosis.


title: Neuromuscular responses of the superficial quadriceps femoris muscles: muscle specific fatigue and inter-individual variability during severe intensity treadmill running.

text: OBJECTIVES This study examined the time course of changes and patterns of responses in electromyographic amplitude (EMG AMP) and EMG mean power frequency (MPF) for the superficial quadriceps muscles during exhaustive treadmill runs within the severe exercise intensity zones (SIZ1 and SIZ2). METHODS The EMG signals for the vastus lateralis (VL), rectus femoris (RF), and vastus medialis (VM) as well as times to exhaustion (Tlim) were recorded in ten runners during two exhaustive treadmill runs (SIZ1 and SIZ2). The composite and individual responses were compared among muscles and between intensities. RESULTS The composite patterns of responses in EMG AMP (linear, quadratic, and cubic increases; r2/R2=0.684-0.848) and EMG MPF (linear, quadratic, and cubic decreases; r2/R2=0.648 - 0.852) for the VL and RF were consistent with neuromuscular fatigue in both zones, but those for the VM were not (quadratic, cubic, and non-significant relationships with responses near baseline). The RF tended to demonstrate greater fatigue (EMG MPF decreased from 80-100% Tlim). There was large inter-individual variability (only 10-60% of responses consistent with composite) in response to fatiguing treadmill running. CONCLUSIONS The current findings support the examination and characterization of neuromuscular fatigue on an intensity, muscle, and subject-by-subject basis.


title: Identification of Japanese encephalitis virus-inducible genes in mouse brain and characterization of GARG39/IFIT2 as a microtubule-associated protein.

text: Several mouse central nervous system genes have been identified that are differentially regulated during Japanese encephalitis virus (JEV) infection, including those which have not been reported to be induced by any other neurotropic virus. Interestingly, approximately 80 % of JEV-inducible genes identified in this study are also induced by Sindbis virus, indicating activation of common host signalling pathways by these two viruses, despite their diverse life cycles. One of these, the glucocorticoid attenuated response gene 39 (GARG39, also known as IFIT2, ISG54 and MuP54) was characterized further. It was demonstrated that GARG39 protein interacts with microtubules in vitro, co-localizes with beta-tubulin in vivo and is enriched in the mitotic spindle of non-neuronal cells undergoing mitosis. While GARG39 was known for a long time as an inflammation-inducible glucocorticoid attenuated protein, its identification as a microtubule-associated protein in this study suggests a possible role for this protein in cell proliferation, virion assembly/transport and microtubule dynamics.


title: Hyperbaric oxygen therapy may be effective to improve hypoxemia in patients with severe COVID-2019 pneumonia: two case reports.

text: Objectives To determine whether hyperbaric oxygen (HBO2) therapy be effective to improve hypoxemia for severe COVID-19 pneumonia patients. Methods Two male patients ages 57 and 64 years old were treated. Each met at least one of the following criteria: shortness of breath; respiratory rate (RR) ≥30 breaths/minute; finger pulse oxygen saturation (SpO2) ≤93% at rest; and oxygen index (P/F ratio: PaO2/FiO2 ≤300 mmHg). Each case excluded any combination with pneumothorax, pulmonary bullae or other absolute contraindications to HBO2. Patients were treated with 1.5 atmospheres absolute HBO2 with an oxygen concentration of more than 95% for 60 minutes per treatment, once a day for one week. Patients' self-reported symptoms, daily mean SpO2 (SO2), arterial blood gas analysis, D-dimer, lymphocyte, cholinesterase (che) and chest CT were conducted and measured. Results For both patients, dyspnea and shortness of breath were immediately alleviated after the first HBO2 treatment and remarkably relieved after seven days of HBO2 therapy. The RR also decreased daily. Neither patient became critically ill. The decreasing trend of SO2 and P/F ratio was immediately reversed and increased day by day. The lymphocyte count and ratio corresponding to immune function gradually recovered. D-dimer corresponding to peripheral circulation disorders and serum cholinesterase, reflecting liver function had improved. Follow-up chest CT showed that the pulmonary inflammation had clearly subsided. Conclusion Our preliminary uncontrolled case reports suggest that HBO2 therapy may promptly improve the progressive hypoxemia of patients with COVID-2019 pneumonia. However, the limited sample size and study design preclude a definitive statement about the potential effectiveness of HBO2 therapy to COVID-2019 pneumonia. It requires evaluation in randomized clinical trials in future.


title: Pharmacological Management and Prevention Of Exacerbations of Chronic Obstructive Pulmonary Disease in Hospitalized Patients.

text: Rapid-acting bronchodilators, systemic corticosteroids, and antibiotics are among the keys to managing exacerbations of chronic obstructive pulmonary disease. Preventing exacerbations should also be a component of therapy for the disease.


SARS in China spreads from Beijing to poorer inland provinces.


title: A global perspective on avian influenza.

text: Global public health security is both a collective aspiration and a mutual responsibility that demands cooperative action at all levels. The expansion of the current H5N1 avian influenza enzootic and its incursion into human health presents a real and significant threat of an influenza pandemic. The world has for the first time an unprecedented opportunity for pandemic preparation. Current global efforts to tackle the H5N1 pandemic threat are centred around the framework of the International Health Regulations (2005) that requires countries to openly share disease intelligence including clinical samples, viruses and epidemiological information. Present international initiatives also seek to establish more equitable allocation and sharing mechanisms for developing countries, of therapeutic resources, public health interventions and other broad-based support in the event of a pandemic. To be sustainable, country preparatory efforts need to be integrated within wider national emergency preparedness frameworks and emphasise the strengthening of basic capacities in disease surveillance, outbreak response and health systems that can respond to a range of public health emergencies. Such capacity building represents permanent investments in health that will have enduring benefits beyond a pandemic. Preparations must also go beyond the health sector; greater promotion of intersectoral cooperation and an adoption of a whole-of-society approach to preparation is recommended. Broad collaboration is vital in addressing the complex challenge posed by influenza to our collective security.


title: Infants tested for hearing loss--United States, 1999-2001.

text: Hearing loss (HL) occurs in one to three of 1,000 live births annually and, when left undetected, can result in developmental delays. To promote communication from birth, Early Hearing Detection and Intervention (EHDI) programs support early identification of infants with HL. Without EHDI programs, the average age of identification for HL is age 1.5-3.0 years, which is past the start of the critical period for optimal language acquisition. In 2001, a total of 48 states/areas with EHDI tracking and surveillance systems (30 funded by CDC) reported the percentage of newborns screened for HL. This report summarizes the results of an analysis of surveillance data for 1999-2001, which indicate that more infants were screened for HL, received diagnostic audiologic evaluations, and were enrolled in early intervention services in 2001 than in 1999 and 2000. Continued development of EHDI surveillance systems should assist states/areas in providing needed services to children with HL.


title: [Epidemiological study on severe acute respiratory syndrome in Guangdong province].

text: OBJECTIVES To understand the epidemiological characteristics of severe acute respiratory syndrome (SARS) outbreaks in some areas of Guangdong province and to provide scientific basis for prevention and control measures against it. METHODS Standardized questionnaire was used on individual cases. Data on the epidemiological characteristics as time, place, persons and aggregation status of SARS cases, development of the epidemics, were analyzed with software EPI 6.0. RESULTS The incidence of SARS in Guangdong province was 1.72/100,000 with case fatality rate as 3.64%. Most cases of SARS occurred between the last ten days of January and the first ten days of February with the peak (61.88% of the patients) occurred in the first ten days of February. As to the distribution of place, Pearl river delta region-economically developed with great number of mobile population-was heavily affected areas (account for 96.66% of the total patients). The majority of patients were young adults and medical staff seemed to be the most affected subgroup (account for 24.9% of the patients in total). Family and hospital aggregation of patients comprised the another two important characteristics of SARS (account for 37.1% of the total patients). CONCLUSION Current knowledge on SARS suggested that it was an air-borne infectious disease with human beings served as the source of infection. The incubation period of the disease was from 1 to 12 days with a median of 4 days. Respiratory secretions and close contact contributed to person-to-person transmission. Most cases were distributed in Pearl river delta region, an area famous for its economic development and heavy flow of mobile population.


title: The -omics era and its impact.

text: OBJECTIVE To review the advances in clinically useful molecular biologic techniques and to identify their applications, as presented at the 12th Annual William Beaumont Hospital DNA Symposium. DATA SOURCES The 7 manuscripts submitted were reviewed and their major findings were compared with literature on the same or related topics. STUDY SELECTION Manuscripts address the use of molecular techniques in the detection of severe acute respiratory syndrome (SARS) and bacterial ribosome mutations, which may lead to ribosome-targeted drug resistance; pharmacogenomics as a clinical laboratory service and example of warfarin dosing using CYP2C9 mutation analysis; definition of the potential of cytosine arabinoside incorporation into DNA to disrupt transcription using an in vitro model of oligonucleotides; use of laser capture microdissection to isolate solid tumor cells free of nontumor cells; and molecular methods used to classify lymphomas. DATA SYNTHESIS Two current issues related to the use of molecular tests in the clinical laboratories are (1) decentralization of molecular-based testing to a variety of nonmolecular laboratories and (2) need for wider acceptance of molecular-based testing through its incorporation in clinical practice guidelines. Molecular methods have had a major impact on infectious disease through the rapid identification of new infectious agents, SARS, and the characterization of drug resistance. Pharmacogenomics identifies the genetic basis for heritable and interindividual variation in response to drugs. The incorporation of the nucleoside analog, cytosine arabinoside, into DNA leads to local perturbation of DNA structure and reduces the ability of transcription factors to bind to their specific DNA binding elements as measured by electrophoretic mobility shift assays. Laser capture microdissection of tumor cells can provide an adequate number of cells for whole genome amplification. Gene expression microassay profiles of various lymphomas have modified classification systems and predict prognosis and response to therapy. CONCLUSIONS The current -omics era will continue to emphasize the use of microarrays and database software for genomic, transcriptomic, and proteomic screening to search for a useful clinical assay. The number of molecular pathologic techniques will expand as additional disease-associated mutations are defined.


title: A serological survey of human coronavirus in pigs of the Tohoku District of Japan.

text: A total of 2496 swine sera from 60 farms in the Tohoku District of Japan was examined for hemagglutination inhibiting (HI) antibodies to human coronavirus (HCV), swine hemagglutinating encephalomyelitis virus (HEV) and bovine coronavirus (BCV). HI antibodies to HCV OC43 strain and HEV 67N strain were highly prevalent with positivity rates of 91.4 and 82.1%, respectively, while the BCV Kakegawa strain was 44.2% positive. Farm D in Miyagi Prefecture showed the highest antibody titers to HCV OC43 strain with geometric mean titers (GMT) of 1:200. These results suggest that pigs might be infected with HCV or an antigenetically related virus as well as HEV.


title: Survival of microorganisms on antimicrobial filters and the removal efficiency of bioaerosols in an environmental chamber.

text: Exposure to bioaerosols causes various adverse health effects including infectious and respiratory diseases, and hypersensitivity. Controlling exposure to bioaerosols is important for disease control and prevention. In this study, we evaluated the efficacies of various functional filters coated with antimicrobial chemicals in deactivating representative microorganisms on filters or as bioaerosols. Tested functional filters were coated with different chemicals that included (i) Ginkgo and sumac, (ii) Ag-apatite and guanidine phosphate, (iii) SiO2, ZnO, and Al2O3, and (iv) zeolite. To evaluate the filters, we used a model ventilation system (1) to evaluate the removal efficiency of bacteria (Escherichia coli and Legionella pneumophila), bacterial spores (Bacillus subtilis spore), and viruses (MS2 bacteriophage) on various functional filters, and (2) to characterize the removal efficiency of these bioaerosols. All experiments were performed at a constant temperature of 25 degrees C and humidity of 50%. Most bacteria (excluding B. subtilis) rapidly decreased on the functional filter. Therefore, we confirmed that functional filters have antimicrobial effects. Additionally, we evaluated the removal efficiency of various bioaerosols by these filters. We used a six-jet collision nebulizer to generate microbial aerosols and introduced it into the environmental chamber. We then measured the removal efficiency of functional filters with and without a medium-efficiency filter. Most bioaerosol concentrations did not significantly decrease by the functional filter only but decreased by a combination of functional and medium-efficiency filter. In conclusion, functional filters could facilitate biological removal of various bioaerosols, but physical removal of these by functional was minimal. Proper use of chemical-coated filter materials could reduce exposure to these agents.


title: Analysis of the role of autophagy in replication of herpes simplex virus in cell culture.

text: The herpes simplex virus type 1 (HSV-1) neurovirulence gene encoding ICP34.5 controls the autophagy pathway. HSV-1 strains lacking ICP34.5 are attenuated in growth and pathogenesis in animal models and in primary cultured cells. While this growth defect has been attributed to the inability of an ICP34.5-null virus to counteract the induction of translational arrest through the PKR antiviral pathway, the role of autophagy in the regulation of HSV-1 replication is unknown. Here we show that HSV-1 infection induces autophagy in primary murine embryonic fibroblasts and that autophagosome formation is increased to a greater extent following infection with an ICP34.5-deficient virus. Elimination of the autophagic pathway did not significantly alter the replication of wild-type HSV-1 or ICP34.5 mutants. The phosphorylation state of eIF2alpha and viral protein accumulation were unchanged in HSV-1-infected cells unable to undergo autophagy. These data show that while ICP34.5 regulates autophagy, it is the prevention of translational arrest by ICP34.5 rather than its control of autophagy that is the pivotal determinant of efficient HSV-1 replication in primary cell culture.


Northern Ireland disease surveillance, April to June 2007.


title: [Emergency department : COVID-19 crisis and organizational aspects].

text: Emergency departments are on the front line in the management of COVID-19 cases, from screening to the initial management of the most severe cases. The clinical presentation of COVID-19 range from non-specific symptoms to adult acute respiratory distress syndrome (ARDS). Diagnosis is based on PCR from a nasopharyngeal swab and emergency treatment rely on oxygen therapy. Patient's orientation (home, hospitalization, admission in intensive care unit) is a central aspect of emergency management. The shift from a strategy of systematic recognition of potential cases to the one of epidemic mitigation required hospital emergency medicine services to implement crisis management measures, to guarantee admission and hospitalization capacity.


Protect your patients, protect your practice: practical risk assessment in the structuring of opioid therapy in chronic pain.


title: Challenges for medical education in Mexico in the time of COVID-19.

text: By April 2, 2020, only 116 days after the description of the first case of SARS-CoV2 virus infection, which causes the COVID-19 disease, 1,014,673 cases and 50,030 deaths have been recorded in 181 countries.1 In the United States, there have been 244,678 cases and 5,911 deaths recorded (in New York State alone, there are 93,053 cases and 2,538 deaths), while in Mexico, 1,378 cases and 37 deaths have been recorded. This number is expected to keep on increasing in both countries. The health system in Mexico, as well as in the rest of the world, will face an enormous problem in the months to come.


title: [A preliminary study of the structure prediction and expression of SARS-CoV spike protein].

text: In this study, the encoding sequences of SARS-CoV spike protein were analyzed by bioinformatics methods, the structural characteristics and functions were forecasted based on available data. It suggests that the fragment of spike glycoprotein (S401-659) may be crucial for viral attachment and may be a major immunodominant epitope. Then the fragment was amplified and subcloned into expression vector pET28a(+) and pPIC9K. These two plasmids pET28a(+)-S and pPIC9K-S were transformed to E.coli strain BL21(DE3)-star and Pichia pastoris, respectively. SDS-PAGE and Western blot analysis showed that the recombinant protein was successfully expressed. The denatured inclusion bodies were purified with Ni-NTA chelate agarose and its purity can reach 90%.


title: Major genetic marker of nidoviruses encodes a replicative endoribonuclease.

text: Coronaviruses are important pathogens that cause acute respiratory diseases in humans. Replication of the approximately 30-kb positive-strand RNA genome of coronaviruses and discontinuous synthesis of an extensive set of subgenome-length RNAs (transcription) are mediated by the replicase-transcriptase, a barely characterized protein complex that comprises several cellular proteins and up to 16 viral subunits. The coronavirus replicase-transcriptase was recently predicted to contain RNA-processing enzymes that are extremely rare or absent in other RNA viruses. Here, we established and characterized the activity of one of these enzymes, replicative nidoviral uridylate-specific endoribonuclease (NendoU). It is considered a major genetic marker that discriminates nidoviruses (Coronaviridae, Arteriviridae, and Roniviridae) from all other RNA virus families. Bacterially expressed forms of NendoU of severe acute respiratory syndrome coronavirus and human coronavirus 229E were revealed to cleave single-stranded and double-stranded RNA in a Mn(2+)-dependent manner. Single-stranded RNA was cleaved less specifically and effectively, suggesting that double-stranded RNA is the biologically relevant NendoU substrate. Double-stranded RNA substrates were cleaved upstream and downstream of uridylates at GUU or GU sequences to produce molecules with 2'-3' cyclic phosphate ends. 2'-O-ribose-methylated RNA substrates proved to be resistant to cleavage by NendoU, indicating a functional link with the 2'-O-ribose methyltransferase located adjacent to NendoU in the coronavirus replicative polyprotein. A mutagenesis study verified potential active-site residues and allowed us to inactivate NendoU in the full-length human coronavirus 229E clone. Substitution of D6408 by Ala was shown to abolish viral RNA synthesis, demonstrating that NendoU has critical functions in viral replication and transcription.


title: Post-SARS psychological morbidity and stigma among general practitioners and traditional Chinese medicine practitioners in Singapore.

text: INTRODUCTION The severe acute respiratory syndrome (SARS) outbreak has been unique in recent history in its rapidity of transmission, its concentration in healthcare settings, and the large number of healthcare workers who have been infected. This study aims to examine the psychological impact of SARS on general practitioners (GPs) and traditional Chinese medicine (TCM) practitioners in Singapore. MATERIALS AND METHODS Two months after the SARS outbreak, all the GPs and TCM practitioners in Singapore were mailed a set of self-reported questionnaires, which included the General Health Questionnaire (GHQ), the Impact of Event Scale-R (IES-R), and a questionnaire to measure the perception of stigma. RESULTS A total of 721 (29%) GPs and 329 (22%) TCM practitioners responded to the survey. Significantly more GPs had worked in SARS affected facilities and had been directly involved in the care of patients with SARS than the TCM practitioners (P <0.001). Those GPs who were directly involved in the care of patients with SARS were significantly more likely to be GHQ case as compared to those not involved in the care of patients with SARS (P = 0.02; OR = 2.9; 95% CI, 1.3-6.3). The mean score of the GHQ somatic, anxiety and social dysfunction subscales were significantly higher in GPs as compared to TCM Practitioners (P <0.001). The GHQ total score as well as the subscales was significantly correlated with the IES-R and stigma subscales (P <0.05). CONCLUSION The fear, uncertainty and stigma caused by SARS are associated with psychological distress among some of the primary healthcare providers in Singapore.


title: Intraepithelial leukocytes contain a unique subpopulation of NK-like cytotoxic cells active in the defense of gut epithelium to enteric murine coronavirus.

text: Initially the intraepithelial leukocytes (IEL) of specific pathogen free (SPF) mice were compared with those of mice held without isolation and were found to differ markedly in total number and distribution of cell surface antigens. The IEL from SPF mice expressed significantly less Thy-1, Lyt-1, and Lyt-2 antigens than their conventional counterparts. The local cell-mediated immune response of mucosal lymphocytes to an enteric murine coronavirus (MHV-Y) was studied in inbred strains of naive SPF mice. A potent in vitro cytotoxic activity was demonstrated by mucosal leukocytes, especially IEL, and spleen cells for MHV-Y-infected syngeneic and allogeneic target cells. The cytotoxicity was not restricted by the major histocompatibility complex. Targets infected with Pichinde virus, an enveloped nonenterotropic virus, were not lysed by these cells. The phenotype of the IEL effector cell was asialo GM1+, Thy-1-, Lyt-1-, Lyt-2-. This cell represents a small subpopulation of the total IEL. After the in vivo administration of anti-asialo GM1 sera, the virus-specific cytotoxic function of the IEL was markedly diminished in in vitro assays, and there was enhanced persistence of virus in gut tissues in vivo. The IEL effector population is defined as a natural killer-like cell that appears to be active in the defense of the gut epithelium to a murine enteric coronavirus.


title: Mouse hepatitis virus infection of the CNS: a model for defense, disease, and repair.

text: Viral infection of the central nervous system (CNS) results in varied outcomes ranging from encephalitis, paralytic poliomyelitis or other serious consequences. One of the principal factors that directs the outcome of infection is the localized innate immune response, which is proceeded by the adaptive immune response against the invading viral pathogen. The role of the immune system is to contain and control the spread of virus within the CNS, and paradoxically, this response may also be pathological. Studies with a neurotropic murine coronavirus, mouse hepatitis virus (MHV) have provided important insights into how the immune system combats neuroinvasive viruses, and have identified molecular and cellular mechanisms contributing to chronic disease in persistently infected mice.


title: A multiplex RT-PCR for the detection of astrovirus, rotavirus, and reovirus in turkeys.

text: This study was undertaken to develop and validate a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) for simultaneous detection of avian rotavirus, turkey astrovirus-2 (TAstV-2), and avian reovirus. Primers targeting the conserved regions of NSP4 gene of avian rotavirus, polymerase gene of TAstV-2, and S4 gene of avian reovirus were used. The position of bands at 630, 802, and 1120 base pairs on agarose gel confirmed the presence of rotavirus, TAstV-2, and reovirus, respectively. This mRT-PCR was found to be specific as no amplification was observed with avian influenza virus, Newcastle disease virus, turkey coronavirus, avian metapneumovirus, and intestinal contents of uninfected turkey poults. Intestinal contents of poults from flocks suspected of exhibiting "poult enteritis syndrome" were pooled and tested. Of the 120 pooled samples tested, 70% were positive for TAstV-2, 45% for avian rotavirus, and 18% for avian reovirus. These three viruses were detected alone or in different combinations. Of the samples tested, 20% were negative for these three viruses, 38% were positive for a single virus (TAstV or rotavirus or reovirus), and 42% were positive for two or three viruses. This single-tube mRT-PCR assay has the potential to serve as a rapid diagnostic method for the simultaneous detection of the three enteric viruses in turkeys.


Diagnosis of enteritis and enterotoxemia due to Clostridium difficile in captive ostriches (Struthio camelus).


title: Characterization of the rubella virus nonstructural protease domain and its cleavage site.

text: The region of the rubella virus nonstructural open reading frame that contains the papain-like cysteine protease domain and its cleavage site was expressed with a Sindbis virus vector. Cys-1151 has previously been shown to be required for the activity of the protease (L. D. Marr, C.-Y. Wang, and T. K Frey, Virology 198:586-592, 1994). Here we show that His-1272 is also necessary for protease activity, consistent with the active site of the enzyme being composed of a catalytic dyad consisting of Cys-1151 and His-1272. By means of radiochemical amino acid sequencing, the site in the polyprotein cleaved by the nonstructural protease was found to follow Gly-1300 in the sequence Gly-1299-Gly-1300-Gly-1301. Mutagenesis studies demonstrated that change of Gly-1300 to alanine or valine abrogated cleavage. In contrast, Gly-1299 and Gly-1301 could be changed to alanine with retention of cleavage, but a change to valine abrogated cleavage. Coexpression of a construct that contains a cleavage site mutation (to serve as a protease) together with a construct that contains a protease mutation (to serve as a substrate) failed to reveal trans cleavage. Coexpression of wild-type constructs with protease-mutant constructs also failed to reveal trans cleavage, even after extended in vitro incubation following lysis. These results indicate that the protease functions only in cis, at least under the conditions tested.


title: Endovascular treatment for acute ischemic stroke patients: implications and interpretation of IMS III, MR RESCUE, and SYNTHESIS EXPANSION trials: A report from the Working Group of International Congress of Interventional Neurology.

text: OBJECTIVE The results of Interventional Management of Stroke (IMS) III, Magnetic Resonance and REcanalization of Stroke Clots Using Embolectomy (MR RESCUE), and SYNTHESIS EXPANSION trials are expected to affect the practice of endovascular treatment for acute ischemic stroke. The purpose of this report is to review the components of the designs and methods of these trials and to describe the influence of those components on the interpretation of trial results. METHODS A critical review of trial design and conduct of IMS III, MR RESCUE, and SYNTHESIS EXPANSION is performed with emphasis on patient selection, shortcomings in procedural aspects, and methodology of data ascertainment and analysis. The influence of each component is estimated based on published literature including multicenter clinical trials reporting on endovascular treatment for acute ischemic stroke and myocardial infarction. RESULTS We critically examined the time interval between symptom onset and treatment and rates of angiographic recanalization to differentiate between "endovascular treatment" and "parameter optimized endovascular treatment" as it relates to the IMS III, MR RESCUE, and SYNTHESIS EXPANSION trials. All the three trials failed to effectively test "parameter optimized endovascular treatment" due to the delay between symptom onset and treatment and less than optimal rates of recanalization. In all the three trials, the magnitude of benefit with endovascular treatment required to reject the null hypothesis was larger than could be expected based on previous studies. The IMS III and SYNTHESIS EXPANSION trials demonstrated that rates of symptomatic intracerebral hemorrhages subsequent to treatment are similar between IV thrombolytics and endovascular treatment in matched acute ischemic stroke patients. The trials also indirectly validated the superiority/equivalence of IV thrombolytics (compared with endovascular treatment) in patients with minor neurological deficits and those without large vessel occlusion on computed tomographic/magnetic resonance angiography. CONCLUSIONS The results do not support a large magnitude benefit of endovascular treatment in subjects randomized in all the three trials. The possibility that benefits of a smaller magnitude exist in certain patient populations cannot be excluded. Large magnitude benefits can be expected with implementation of "parameter optimized endovascular treatment" in patients with ischemic stroke who are candidates for IV thrombolytics.


SARS: make no mistake--there will be a next time.


Molecular aspects of the relationship of transmissible gastroenteritis virus (TGEV) with porcine respiratory coronavirus (PRCV).


title: In vitro reproduction of device deformation leading to thrombotic complications and failure of flow diversion.

text: Flow diverters (FDs) are increasingly used for complex intracranial aneurysms. As these self-expanding devices are deployed across an aneurysm neck, they can undergo deformations. The potential clinical consequences of FD deformations remain unclear. We describe an immediate thrombotic complication attributed to a stereotypical stenotic deformation of an FD extremity that can occur when landing zones are of insufficient length. This case is supplemented with in vitro studies showing the relationship between i) the length of the landing zones and ii) discrepancies between the diameter of the device and recipient vessel, and the severity of FD stenosis. In vitro, a shorter landing zone was associated with a progressive stenotic deformation of the terminal ends of all FDs studied. This deformation was more pronounced when the diameter of the device was oversized compared to the size of the recipient tube. In our clinical case, the presence of this deformation led to an immediate thrombotic complication, requiring deployment of a second stent to correct the observed stenosis. In addition, treatment failure ultimately led to a fatal rupture, a failure that can be explained by residual flows through a more porous transition zone, another characteristic FD deformation which occurs when they are oversized as compared to the parent vessel, but free to expand at the level of the aneurysm. Proper selection of device diameter and length of the landing zone is important, and may decrease the incidence of deformation-related complications.


title: Factors associated with pathogen seroprevalence and infection in Rocky Mountain cougars.

text: Serological and genetic material collected over 15 years (1990-2004) from 207 cougars (Puma concolor) in four populations in the Rocky Mountains were examined for evidence of current or prior exposure to feline immunodeficiency virus (FIV), feline parvovirus (FPV), feline coronavirus (FCoV), feline calicivirus (FCV), canine distemper virus (CDV), feline herpesvirus (FHV), and Yersinia pestis. Serologic data were analyzed for annual variation in seroconversions to assess whether these pathogens are epidemic or endemic in cougars, and to determine whether family membership, age, sex, or location influence risk of exposure. FIV and FPV were clearly endemic in the studied populations, whereas exposure to FCoV, FCV, CDV, and Y. pestis was more sporadic. No evidence was found for FHV. Age was the most consistent predictor of increased exposure risk, often with no other important factors emerging. Evidence for transmission within family groups was limited to FIV and FCoV, whereas some indication for host sex affecting exposure probability was found for FIV and Y. pestis. Overall, cougar populations exhibited few differences in terms of pathogen presence and prevalence, suggesting the presence of similar risk factors throughout the study region.


title: Data mining pathogen genomes using GeneOrder and CoreGenes and CGUG: gene order, synteny and in silico proteomes.

text: Sequence databases are growing exponentially due to 'next generation' DNA analysers and applications of these data. Databases include multiple sequences of previously sequenced organisms, particularly ones of consequence to human health. Applications are limited by tools available to mine them, particularly user-friendly tools that are useful for bench researchers. GeneOrder, CoreGenes and CGUG are web-based 'on-the-fly' tools that examine gene order and synteny, as well as proteomes for comparative genomics and for drug discovery and design targets. CoreGenes (CGUG) now allows analysis of genomes ranging up to 1.9 megabases. Many of these small genome bacteria have impacts on human health.


title: Seeds as a production system for molecular pharming applications: status and prospects.

text: The production of recombinant proteins in seeds is achieved by driving transgene expression using promoters and protein targeting sequences derived from genes encoding abundant seed storage proteins. This approach is advantageous because high yields, stability and containment are conferred by the accumulation of recombinant proteins in specialized storage compartments such as protein bodies and protein storage vacuoles. Seeds are particularly suitable for the production of pharmaceutical proteins in developing country settings because they reduce the costs of production and distribution by avoiding the need for fermenter-based production capacity and a cold chain for storage and distribution, thus increasing access to critical medicines for the poor in rural areas. Seeds are also ideal for the production of oral vaccine antigens, because encapsulation within the seed provides protection that prolongs exposure to the gastric immune system and thus increases the potency of the immune response. In this review we discuss the current state of the art in seed-based molecular pharming and the future potential of production platforms based on seeds.


title: Viral enteritis of calves.

text: Calf diarrhoea is an important cause of economic loss. The aetiology of the disease is considered with particular reference to the role of viruses. Although many microbial organisms have been associated with the disease, there is doubt concerning their true role as causative agents. Two viruses, neonatal calf diarrhoea reovirus-life agent and calf coronavirus, have been discovered recently which appear to play an important role in many cases of diarrhoea and the evidence for considering them to be primary causes of the disease is discussed.


title: Chest radiographic manifestations of severe acute respiratory syndrome in health care workers: the Toronto experience.

text: OBJECTIVE The purpose of this article is to describe the chest radiographic manifestations of severe acute respiratory syndrome (SARS) in previously uninfected health care workers during the early stages of an outbreak in Toronto, Canada. CONCLUSION The study group was composed of 13 patients from a single institution. Three distinct chest radiographic patterns were observed: focal peripheral air-space disease at presentation with gradual resolution (most common pattern, 10/13 patients), normal findings on chest radiography at presentation followed by focal air-space disease (2/13 patients), and normal findings on chest radiography at presentation followed by a "round" pneumonia pattern (1/13 patients). There was no evidence of pleural reaction, lymphadenopathy, or interstitial changes.


Comparison of replicase localization in different types of mouse hepatitis virus (MHV)-infected cells.