title: Influenza as a model system for studying the cross-species transfer and evolution of the SARS coronavirus.

text: Severe acute respiratory syndrome coronavirus (SARS-CoV) moved into humans from a reservoir species and subsequently caused an epidemic in its new host. We know little about the processes that allowed the cross-species transfer of this previously unknown virus. I discuss what we have learned about the movement of viruses into humans from studies of influenza A, both how it crossed from birds to humans and how it subsequently evolved within the human population. Starting with a brief review of severe acute respiratory syndrome to highlight the kinds of problems we face in learning about this viral disease, I then turn to influenza A, focusing on three topics. First, I present a reanalysis of data used to test the hypothesis that swine served as a "mixing vessel" or intermediate host in the transmission of avian influenza to humans during the 1918 "Spanish flu" pandemic. Second, I review studies of archived viruses from the three recent influenza pandemics. Third, I discuss current limitations in using molecular data to study the evolution of infectious disease. Although influenza A and SARS-CoV differ in many ways, our knowledge of influenza A may provide important clues about what limits or favours cross-species transfers and subsequent epidemics of newly emerging pathogens.


title: Coronavirus IBV: relationships among recent European isolates studied by limited proteolysis of the virion glycopolypeptides.

text: After the year 1978, strains of avian infectious bronchitis virus (IBV) were isolated in the Netherlands and the UK which were assigned by neutralisation tests to four new serotypes (arbitrarily designated A, B, C and D in this communication) distinct from the long recognised US serotypes. We have labelled, with 35S-methionine, the structural polypeptides of 12 of the European isolates during growth in de-embryonated eggs. The S2, spike-anchoring, polypeptide of isolates of all four serotypes had a molecular weight of 87 000 (87K) whereas the US IBV-M41 strain had S2 of 84K. When the isolates were grouped according to the molecular weight of the other spike glycopolypeptide, S1, and the membrane (M) glycopolypeptide four groups emerged which corresponded to the serotypes based on neutralisation tests. Serotypes A (isolates D207, 1, 5 and 6) and B (isolates 7, 8 and D3896) had S1 of 91.5K while serotypes C (isolates 9 and D3128) and D (isolates D212 and D1466) had S1 of 90K. The M of serotypes A and D had a molecular weight of 30K while that of serotypes B and C was 27K. Chymotrypsin and protease V8 were used for limited proteolysis, the hydrolysis of S1 giving the greatest discrimination between serotypes. After proteolysis of S1 the UK isolates of serotype A gave identical profiles which were very similar to the Dutch serotype A isolate D207. Isolates of serotype B gave S1 profiles very similar to those of serotype A, as did D274 which is serologically related to both groups A and B. Serotypes C and D were distinguishable from each other and from serotypes A and B on the basis of the S1 peptide profiles. These results strengthen the view that the UK and Dutch isolates are closely related and that serotypes A and B are more closely related to each other than to either group C or D serotypes.


Hepatitis C virus (HCV) genotypes and disease progression in HIV/HCV-coinfected patients.


title: Robotic Transhiatal Esophagectomy in a Community Hospital: Evolution of Technique.

text: Esophageal cancer is an uncommon but highly lethal disease. Surgical resection is the gold standard of treatment for early-stage disease. Traditional surgical approach entailed significant convalescence, hospital stay, and morbidity and mortality. Transhiatal esophagectomy (THE) involves blind dissection of the esophagus with minimal mediastinal lymphadenectomy. Integration of robotic surgery is an alternate platform for minimally invasive approach while maintaining safety and following oncologic principles. We review our technique for minimally invasive THE using robotic technology, demonstrating the safety and efficacy of robotic technology surgery. We present a retrospective review of a single surgeon's data of patients treated with robotic-assisted THE, with a chart review to evaluate pathology, adequacy of surgical resection, nodal harvest, and perioperative course. Robotic THE (rTHE) shows promise as a valid option for esophageal resection, including premalignant and advanced stages of cancer. Adequate transhiatal mediastinal nodal resection can be performed with the robot.


title: Colour-coded duplex sonography of the testes of dogs.

text: Colour-coded duplex sonography was used to examine the testes of 42 dogs whose testes were normal on clinical examination. With colour Doppler, the blood flow in the supratesticular region could be displayed clearly in 70 per cent of the cases, independent of the dog's age, bodyweight, pulse rate and the volume of its testes. The marginal artery could be displayed clearly in 56 per cent of the cases, and the clarity was correlated with bodyweight (P<0.001), testicular volume (P<0.001), and pulse rate (P<0.001). The intratesticular blood flow could not be discerned in 42 per cent of the cases. With pulsed Doppler, the mean resistive index (RI) in the supratesticular region was 0.57, and the mean pulsatility index (PI) was 1.00. In the marginal artery, the mean RI was 0.49, and mean PI was 0.78. In the intratesticular arteries, the mean RI was 0.47, and the mean PI was 0.75. At all three sites both indices were independent of age, bodyweight, pulse rate and testicular volume.


title: Isolation and characterization of a coronavirus from pigeons with pancreatitis.

text: OBJECTIVE To identify and partially characterize a coronaviruslike virus isolated from naturally infected pigeons. ANIMALS 50 specific pathogen-free (SPF) embryonated chicken eggs, 30 White Leghorn SPF chickens, and 12 clinically normal pigeons. PROCEDURES Pancreatic tissue specimens from sick pigeons were inoculated into SPF embryonated chicken eggs for viral isolation and investigation of morphologic and hemagglutinating properties of the isolate, called PSH050513. Furthermore, virulence studies in SPF chickens and experimental pigeons were performed. The spike (S) glycoprotein gene of PSH050513 was further sequenced and analyzed. RESULTS PSH050513 was isolated and identified from the experimentally infected pigeons by a routine method, which was in accordance with Koch's postulates. The complete S protein (1,167 amino acids) was compared with published S protein sequences of other avian and mammalian coronaviruses. A high degree of sequence identity (79.3% to 99.6%) was observed between the S protein sequence of PSH050513 and published sequences of avian infectious bronchitis virus (IBV); only limited identity (< 37.8%) was observed with turkey coronavirus and mammalian coronaviruses. Furthermore, when the virus was inoculated into SPF chickens, pancreatitis developed. CONCLUSIONS AND CLINICAL RELEVANCE PSH050513 has been tentatively identified as a novel member of group 3 coronaviruses that have close genetic relationships with IBV strains.


title: Effect of Beetroot Juice on Moderate-Intensity Exercise at a Constant Rating of Perceived Exertion.

text: Dietary nitrate supplementation has been shown to reduce oxygen consumption at a fixed work rate. We questioned whether a similar effect would be observed during variable work rate exercise at a specific rating of perceived exertion (RPE), as is commonly prescribed for aerobic training sessions. Using a double-blind, placebo controlled, crossover design, ten females (25 ± 3 years; VO2peak 37.1 ± 5.3 ml/kg/min) performed two 20-min cycle ergometer trials at a constant RPE of 13 (somewhat hard) 2.5 hours following ingestion of 140 ml of concentrated beetroot juice (12.9 mmol nitrate), or nitrate-depleted placebo. Performance was measured in terms of total VO2 (L) consumed and total mechanical work (kJ) accomplished across each trial. Following each experimental trial, subjects rode at 75W for an additional 5 min to determine the effect of beetroot juice on fixed work rate exercise. Coefficients of variation in total VO2 (L) and work performed (kJ) during the RPE 13 clamp trials were 8.2 and 9.5%, respectively. Consumption of beetroot juice did not affect total VO2 or work performed during RPE 13 exercise, but lowered resting systolic blood pressure by ~5 mmHg (P=0.041) and oxygen consumption at 75W by ~4% (P=0.048), relative to placebo. Since the effect of beetroot juice on oxygen consumption is small and may be masked by daily variability during self-regulated exercise, it is unlikely to have a notable effect on daily training.


Plaque assay of bovine coronavirus in BEK-1 cells.


title: Feline viruses in wildcats from Scotland.

text: Few data are available on the prevalence of feline viruses in European wildcats (Felis silvestris). Previous surveys have indicated that wildcats may be infected with the common viruses of domestic cats, apart from feline immunodeficiency virus (FIV). In the present study, 50 wildcats trapped throughout Scotland (UK) between August 1992 and January 1997 were tested for evidence of viral infection. All were negative for FIV by several serological or virological methods. By contrast, 10% of the cats were positive for feline leukemia virus (FeLV) antigen and infectious virus was isolated from 13% of a smaller subset. Of the wildcats tested for respiratory viruses, 25% yielded feline calicivirus (FCV) and although no feline herpesvirus was isolated, 16% of the samples had neutralizing antibodies to this virus. Antibodies to feline coronavirus (FCoV) were found in 6% of samples. Feline foamy virus (FFV) was an incidental finding in 33% of samples tested. This study confirms that wildcats in Scotland are commonly infected with the major viruses of the domestic cat, except for FIV.


title: Treatment of the common cold with unrefined echinacea. A randomized, double-blind, placebo-controlled trial.

text: BACKGROUND Echinacea preparations are widely used to treat the common cold. OBJECTIVE To assess the efficacy of dried, encapsulated, whole-plant echinacea as early treatment for the common cold. DESIGN Randomized, double-blind, placebo-controlled community-based trial. SETTING University of Wisconsin-Madison, Madison, Wisconsin. PARTICIPANTS 148 registered students with common colds of recent onset. INTERVENTION An encapsulated mixture of unrefined Echinacea purpurea herb (25%) and root (25%) and E. angustifolia root (50%) taken in 1-g doses six times on the first day of illness and three times on each subsequent day of illness for a maximum of 10 days. MEASUREMENTS Severity and duration of self-reported symptoms of upper respiratory tract infection. RESULTS No statistically significant differences were detected between the echinacea and placebo groups for any of the measured outcomes. Trajectories of severity over time were nearly identical in the two groups. Mean cold duration was 6.01 days in both groups as a whole, 5.75 days in the placebo group, and 6.27 days in the echinacea group (between-group difference, -0.52 day [95% CI, -1.09 to 0.22 days]). After controlling for severity and duration of symptoms before study entry, sex, date of enrollment, and use of nonprotocol medications, researchers found no statistically significant treatment effect (adjusted hazard ratio, 1.24 [CI, 0.86 to 1.78]). Multivariable regression models assessing severity scores over time failed to detect statistically significant differences between the echinacea and placebo groups. CONCLUSION Compared with placebo, unrefined echinacea provided no detectable benefit or harm in these college students who had the common cold.


title: Scanning electron microscopy of abomasium and intestine of gnotoxenic calves infected either with rotavirus, coronarivus or enteropathogenic Escherichia coli or with rotavirus and E. coli.

text: Neonatal calf diarrhoea induced with several agents of infection was studied by scanning electron microscopy. In a gnotoxenic calf infected with E. coli K99+ Ent+, slight lesions of the small intestine were observed and desquamation or puffiness of microvilli occurred. In rotavirus-infected calves, the abomasum was covered with abudant mucous film and appeared to be desquamated. In the small intestine, no desquamation of epithelium was observed. Inoculation of the rotavirus and E. coli induced severe diarrhoea. The whole digestive tract, even the abomasum and colon, was eroded. Coronavirus induced marked lesions in all levels of the intestine. These results demonstrate unequivocally the pathogenic properties of the three infectious agnets, the synergistic effect of E. coli and rotavirus. Furthermore, the importance of the abomasum in neonatal diarrhoea is emphasized.


title: The Gastroenterological Society of Australia.

text: In this review, a summary of the objectives and activities of The Gastroenterological Society of Australia is given. It illustrates the multidisciplinary and multi-functional roles of a society that is confident and active in supporting and reflecting the views of its members in the pursuit of excellence in clinical practice, research and education in the various areas of digestive health.


title: Vaccinia virus entry, exit, and interaction with differentiated human airway epithelia.

text: Variola virus, the causative agent of smallpox, enters and exits the host via the respiratory route. To better understand the pathogenesis of poxvirus infection and its interaction with respiratory epithelia, we used vaccinia virus and examined its interaction with primary cultures of well-differentiated human airway epithelia. We found that vaccinia virus preferentially infected the epithelia through the basolateral membrane and released viral progeny across the apical membrane. Despite infection and virus production, epithelia retained tight junctions, transepithelial electrical conductance, and a steep transepithelial concentration gradient of virus, indicating integrity of the epithelial barrier. In fact, during the first four days of infection, epithelial height and cell number increased. These morphological changes and maintenance of epithelial integrity required vaccinia virus growth factor, which was released basolaterally, where it activated epidermal growth factor 1 receptors. These data suggest a complex interaction between the virus and differentiated airway epithelia; the virus preferentially enters the cells basolaterally, exits apically, and maintains epithelial integrity by stimulating growth factor receptors.


title: Reovirus binding to cell surface sialic acid potentiates virus-induced apoptosis.

text: Reovirus induces apoptosis in cultured cells and in vivo. Genetic studies indicate that the efficiency with which reovirus strains induce apoptosis is determined by the viral S1 gene, which encodes attachment protein sigma1. However, the biochemical properties of sigma1 that influence apoptosis induction are unknown. To determine whether the capacity of sigma1 to bind cell surface sialic acid determines the magnitude of the apoptotic response, we used isogenic reovirus mutants that differ in the capacity to engage sialic acid. We found that T3SA+, a virus capable of binding sialic acid, induces high levels of apoptosis in both HeLa cells and L cells. In contrast, non-sialic-acid-binding strain T3SA- induces little or no apoptosis in these cell types. Differences in the capacity of T3SA- and T3SA+ to induce apoptosis are not due to differences in viral protein synthesis or production of viral progeny. Removal of cell surface sialic acid with neuraminidase abolishes the capacity of T3SA+ to induce apoptosis. Similarly, incubation of T3SA+ with sialyllactose, a trisaccharide comprised of lactose and sialic acid, blocks apoptosis. These findings demonstrate that reovirus binding to cell surface sialic acid is a critical requirement for the efficient induction of apoptosis and suggest that virus receptor utilization plays an important role in regulating cell death.


title: Clinical characteristics of fatal patients with severe acute respiratory syndrome in a medical center in Taipei.

text: BACKGROUND The clinical spectrum of severe acute respiratory syndrome (SARS) varies widely in the way that asymptomatic carriers are believed to exist in the community. Still there are severe forms of illness in which the patients deteriorate unexpectedly within hours. This study delineates clinical characteristics of such fatal cases of SARS for the purpose of identifying patients with poor outcome. METHOD Patients with the diagnosis of probable or suspected SARS admitted to the Taipei Veterans General Hospital during the period from March 26, 2003 to May 25, 2003 were included. The medical records of fatal cases were retrospectively reviewed. RESULTS During the study period, thirty-six probable cases and 17 suspected cases of SARS were identified. Eight probable but none of the suspected cases died from acute respiratory distress syndrome (ARDS) with multiple organ dysfunction after a median of 6-day hospital stay (range, 1-30 days). All but 2 patients acquired the infection nosocomially. Of the fatal cases, four were males and 4 females, with the median age of 65 years (range, 29-76 yrs). All except 3 had co-morbid conditions, such as hypertension, diabetes mellitus, coronary artery disease or chronic obstructive pulmonary disease. The lactate dehydrogenase (LDH) values were abnormal (> 200 U/L) in all patients. The chest radiograph of these fatal cases consistently showed multifocal infiltration over the unilateral or bilateral lobe of the lung. CONCLUSIONS Patients of SARS who had advanced age, co-morbid conditions, highly elevated LDH and multifocal infiltration over chest radiograph should be closely monitored and actively treated.


Implications of the SARS outbreak for Canadian emergency departments.


title: Viral respiratory infections due to rhinoviruses: current knowledge, new developments.

text: Viral respiratory infections (VRIs) are among the most common reasons for which primary care providers are consulted. VRIs due to rhinoviruses-the most commonly implicated etiologic agent-constitute a syndrome characterized by signs and symptoms of a cold. Rhinoviruses have been implicated in respiratory tract illnesses such as sinusitis and otitis media, as well as lower respiratory complications in high-risk populations. Most patients treat VRI with over-the-counter remedies that have been demonstrated to produce marginal clinical benefits. The development of novel antiviral agents has intensified interest in VRIs. Pleconaril, a capsid-function inhibitor currently under FDA review, has been shown in clinical trials to reduce the duration and severity of rhinovirus VRIs. By targeting the cause of illness, antiviral agents represent an opportunity to reduce the substantial clinical burden of VRI. Furthermore, effective therapies can potentially reduce inappropriate antibiotic use for viral infections.


title: [An outbreak of SARS in Dongcheng District, Beijing during March to June 2003].

text: OBJECTIVE To describe epidemiological characteristics of an outbreak of severe acute respiratory syndrome (SRAS) in Dongcheng District, Beijing during March to June 2003. METHODS Data of SARS cases notified from Dongcheng District Center for Disease Control and Prevention (CDC) and supplemented by other channels were collected. All data were input into dataset files by Microsoft Excel-2000 software and analyzed with SPSS version 10.0 software. RESULTS Totally, 572 cases notified were collected during this period in Dongcheng District, Beijing, and 99 of them were excluded from SARS, because of diagnosis of common cold, pneumonia, measles, or rubella, etc. Actually, 473 SARS cases were analyzed. Attack rate of SARS in permanent residents of Dongcheng was 28.3 per 100,000. Forty-one of them died, with a case-fatality ratio of 8.7%. Outbreak of SARS in Dongcheng District started on March 14, with a peak during mid- and late April, and dropped from May 5, 2003. Persons were all susceptible to SARS, with the highest proportion at ages of 20-50 years, which accounted for 68.7% of the total cases. Average age of the patients at their onset was 40.7 years. No gender difference in SARS cases was found. Number of SARS cases in health care workers (HCWs) accounted for 18.0% and that in the retired persons accounted for 15.4% of the total cases. Cases occurred in all 10 sub-districts of Dongcheng. Totally, 230 of the 572 notified cases (40.2%) hospitalized at local hospitals under the jurisdiction of Dongcheng District. Eighteen of 85 cases of SARS occurred in health care workers of local hospitals, accounting for 4.5% of the total number of health care workers working at the wards caring for SARS patients or at fever clinics. There were 34.7% of SARS cases without any contact histories before their onset. Familial aggregation phenomena were observed in 41.8% of the cases and 18.1% of households. And, 7.4% (attack rate) in those exposed to SARS cases suffered the illness during the periods of quarantine. CONCLUSIONS SARS appeared outbreak in Dongcheng District, Beijing during March to June 2003. People were all susceptible to SARS, which mainly threatened the young adults and the middle-aged, as well as health care workers and the retired workers. Main mode of transmission was exposure to SARS patients in a near distance at hospitals or families. Prevention and control of SARS should be focused on early isolation of patients and quarantine for the contacts. Current available measures to prevent and control SARS proved effective.


title: Severe acute respiratory syndrome: radiographic review of 40 probable cases in Toronto, Canada.

text: PURPOSE To review radiographic findings of patients with probable severe acute respiratory syndrome (SARS) who were seen at a University of Toronto (Ontario, Canada) teaching hospital. MATERIALS AND METHODS Findings were reviewed for 40 patients who fulfilled the World Health Organization criteria for probable SARS. A template was designed for the analysis of each serial radiograph to observe patterns and distribution of disease, interval changes, and complications. The majority of radiographs were anteroposterior views. A clinical database of these patients was also collected for clinical-radiologic comparison. RESULTS The mean age of the patients (18 male, 22 female) was 42.7 years. Patients had a normal chest radiograph and focal, multifocal, and/or bilateral consolidation. The pattern of consolidation tended to be peripheral and poorly marginated and involved middle and lower lung zones. The serial sequence fell into two major subgroups, which correlated closely with clinical outcome. Consolidation in one group cleared within a matter of days, while the second group went on to develop rapid and extensive bilateral pneumonia, with a prolonged hospital stay. Subsegmental atelectasis and pleural complications were rarely observed. CONCLUSION SARS pneumonia can manifest as focal peripheral consolidation that clears relatively quickly and does not cause secondary complications or that progresses to bilateral consolidation and a more protracted clinical course.


title: Preclinical evaluation of antimicrobial efficacy and biocompatibility of a novel bacterial barrier dressing.

text: UNLABELLED  Wounds that become infected can lead to devastating consequences for patients, resulting in substantially increased healthcare costs. Bacterial barrier dressings are a first line of protection against developing wound infections. Most bacterial barrier dressings contain microbicidal chemicals (eg, silver ions, iodine, chlorhexidine) that are released from the dressings, which can be toxic to wound cells. A need exists for cost effective bacterial barrier dressings that absorb wound exudate and do not release toxic materials into the wound or increase the risk for developing bacterial resistance to the microbicidal chemical. The present study reports the development and properties of a novel bacterial barrier dressing that meets these needs. METHODS A high molecular weight (~250 k Daltons) polymer containing a high density of quaternary amines (polydiallyldimethylammonium chloride [polyDADMAC]) was permanently bonded onto cellulose fibers (gauze). Microbicidal and mammalian cell cytotoxicity tests were conducted using standard methods. Development of bacterial resistance to the microbicidal fibers was assessed over 10 passages. RESULTS The polyquat-treated bacterial barrier gauze dressing (BIOGUARD [BBD]) had high microbicidal activity even in the presence of proteinaceous fluid against a wide range of Gram-positive and Gram-negative bacteria. The BBD dressing passed all mammalian cell toxicity tests due to the non-leaching of the bactericidal polymer. Furthermore, the BBD dressing did not demonstrate any ability to induce bacterial resistance in selection vector testing. CONCLUSION This novel dressing featuring a bound microbicide offers another choice for wound caregivers to provide patients with an antimicrobial barrier dressing safe enough for prophylactic use to protect against wound infections.


title: COPD and cognitive impairment: the role of hypoxemia and oxygen therapy.

text: BACKGROUND several studies have shown an association between chronic obstructive pulmonary disease (COPD) and cognitive impairment. These studies have been limited by methodological issues such as diagnostic uncertainty, cross-sectional design, small sample size, or lack of appropriate referent group. This study aimed to elucidate the association between COPD and the risk of cognitive impairment compared to referent subjects without COPD. In patients with established COPD, we evaluated the impact of disease severity and impairment of respiratory physiology on cognitive impairment and the potential mitigating role of oxygen therapy. METHODS we used the Function, Living, Outcomes and Work (FLOW) cohort study of adults with COPD (n = 1202) and referent subjects matched by age, sex, and race (n = 302) to study the potential risk factors for cognitive impairment among subjects with COPD. Cognitive impairment was defined as a Mini-Mental State Exam score of <24 points. Disease severity was using Forced Expiratory Volume in one second (FEV(1)); the validated COPD Severity Score; and the BMI (Body Mass Index), Obstruction, Dyspnea, Exercise Capacity (BODE) Index. Multivariable analysis was used to control for confounding by age, sex, race, educational attainment, and cigarette smoking. RESULTS COPD was associated with a substantive risk of cognitive impairment compared to referent subjects (odds ratio [OR] 2.42; 95% confidence interval [CI] 1.043-6.64). Among COPD patients, none of the COPD severity measures were associated with the risk of cognitive impairment (P > 0.20 in all cases). Low baseline oxygen saturation was related to increased risk of cognitive impairment (OR for oxygen saturation ≤88% (OR 5.45; 95% CI 1.014-29.2; P = 0.048). Conversely, regular use of supplemental oxygen therapy decreased the risk for cognitive impairment (OR 0.14; 95% CI 0.07-0.27; P < 0.0001). CONCLUSION COPD is a major risk factor for cognitive impairment. Among patients with COPD, hypoxemia is a major contributor and regular use of home oxygen is protective. Health care providers should consider screening their COPD patients for cognitive impairment.


title: Serological and molecular evidence of enterovirus infection in patients with end-stage dilated cardiomyopathy.

text: OBJECTIVE To study the relative diagnostic value of enterovirus-specific molecular biological and serological assays in patients with end-stage dilated cardiomyopathy, and to investigate the possible role of other cardiotropic viruses in dilated cardiomyopathy. DESIGN Analysis of recipient myocardial tissue and serum from patients with dilated cardiomyopathy and controls undergoing cardiac transplantation for end-stage cardiac disease. SETTING University virology department and transplantation unit. METHODS Reverse transcriptase-polymerase chain reaction and nucleotide sequence analysis of myocardial RNA and DNA; enterovirus-specific in situ hybridization; enterovirus-specific immunoglobulin M detection. RESULTS Enterovirus RNA was detected in myocardial tissue from only a small proportion of (five of 75) hearts. However, although enterovirus-specific immunoglobulin M responses were detected in 22 (28%) of 39 controls patients, a significantly higher prevalence was observed among patients with dilated cardiomyopathy (22 (56%) of 39 patients; P < 0.005). All enteroviruses detected in myocardium showed greatest nucleotide sequence homology with coxsackievirus type B3. Detection of enterovirus RNA in myocardium by the polymerase chain reaction and by in situ hybridisation gave comparable results. Other potentially cardiotropic virus genomes, including human cytomegalovirus, influenzaviruses, and coronaviruses were not detected in myocardium. CONCLUSION This study found that enterovirus-specific immunoglobulin M responses provided the strongest evidence of enterovirus involvement in patients with end-stage dilated cardiomyopathy. However, the high background prevalence of these responses limits their diagnostic value. The finding that enteroviruses detected in myocardium were coxsackievirus type B3 accords with recent findings in patients with acute myocarditis, and indicates that this serotype is the major cardiotropic human enterovirus.


title: Use of IgY antibody to recombinant avian reovirus σC protein in the virus diagnostics.

text: Avian reovirus (ARV) is an important agent of several diseases causing considerable losses in poultry farming. An outer capsid protein (σC) of ARV, is known as a virus-cell attachment protein essential for virus infectivity. In this study, the σC gene of ARV was cloned and expressed in Escherichia coli. The expressed recombinant protein was used as immunogen for raising a specific IgY antibody in laying hens. At 14 weeks post immunization, the antibody titers in serum and egg yolk reached 302,000 and 355,000, respectively. The IgY antibody was capable to neutralize ARV in BHK-21 cells and it strongly reacted in ELISA with ARV but not with heterologous viruses. The IgY antibody detected ARV in field samples of infected animal tissues in dot blot assay. These results suggest that an efficient, economic and rapid diagnostics of ARV can be performed routinely using the IgY antibody against a recombinant ARV σC protein.


title: Severe acute respiratory syndrome.

text: The mysterious severe acute respiratory syndrome (SARS) that has originated from the southern Chinese province of Guangdong appears to be a major public health threat and medical challenge. Laboratory studies of SARS patients in a number of countries identified the etiologic agent being a novel member of coronaviridae. High RNA concentrations of this virus in sputum make it as a highly infectious agent. Low concentrations of viral genome are also detectable in feces. Coronaviruses are ubiquitos. They cause disease in many animals including pigs, cattle, dogs, cats, and chickens. These viruses have been associated with upper respiratory infections and sometimes pneumonia in humans. SARS presents with fever, cough, malaise, dyspnea, and hypoxemia. Chest radiographs from affected regions are associated with progressive airway disease. Clinical laboratory features of SARS include lymphopenia, thrombocytopenia, and elevated lactate dehydrogenase levels. Currently, there is no FDA approved pharmacologic treatment for SARS. To date, no convincing clinical data is available for treatment of SARS with ribavirin. While there are some controversies about the use of systemic corticosteroids, Martin et al, in this issue of MSM, present their views on the use of pentoxyfylline (PTX) as a potential agent to be considered for SARS treatment. Finally, our analytical approach to the risks of SARS will certainly enable us to


title: Clinical brief: severe acute respiratory syndrome (SARS).

text: The World Health Organization (http:/www.who.int/en/) and Centers for Disease Control and Prevention (http://www.cdc.gov) websites are updated daily as new information is learned about the Severe Acute Respiratory Syndrome (SARS) phenomena. The transmission of SARS appears to occur predominantly through direct contact with infectious material. This includes dispersed respiratory droplets, making touching contaminated objects a potential concern. Occupational health nurses must educate themselves and their employees about SARS to allow for accurate and timely sharing of information.


Diaphragmatic Crural Eversion: Mid-Term Data of a Novel Technique to Optimize Hiatoplasty during Laparoscopic Repair of Hiatal Hernia.


title: Biological and molecular differentiation between coronaviruses associated with neonatal calf diarrhoea and winter dysentery in adult cattle.

text: Cytopathic coronaviruses were isolated in HRT-18 cells from bloody faecal samples collected from cows in Québec dairy herds with classical winter dysentery (WD). The formation of polykaryons in the infected cell cultures was found to be dependent on the presence of trypsin in the medium. Virus identification was confirmed by indirect immunofluorescence and indirect protein A-gold immunoelectron microscopy using rabbit hyperimmune serum, as well as monoclonal antibodies directed against the spike (S) and hemagglutinin-esterase (HE) glycoproteins of the prototype Mebus strain of bovine coronavirus (BCV-Meb). Four WD isolates differed from BCV-Meb by their ability to agglutinate rat erythrocytes at 4 and 37 degrees C, their higher receptor destroying enzyme activity, but lower acetylesterase activity. The WD isolates were serologically indistinguishable from the reference BCV-Meb strain by virus neutralization and Western immunoblotting, but could be differentiated by hemagglutination-inhibition. Sequence analysis of the PCR-amplified HE gene of a plaque-purified WD isolate (BCQ-2590) revealed sufficient number of nucleotide and amino acid substitutions which may explain this antigenic variability.


title: Genomic organization and expression of the 3' end of the canine and feline enteric coronaviruses.

text: The genomic organization at the 3' end of canine coronavirus (CCV) and feline enteric coronavirus (FECV) was determined by sequence analysis and compared to that of feline infectious peritonitis virus (FIPV) and transmissible gastroenteritis virus (TGEV) of swine. Comparison of the latter two has previously revealed an extra open reading frame (ORF) at the 3' end of the FIPV genome, lacking in TGEV, now designated ORF 6b. Both CCV and FECV possess 6b-related ORFs. The CCV ORF 6b is colinear with that of FIPV, but the predicted amino acid sequences are only 58% identical. The FECV ORF 6b contains a large deletion compared to that of FIPV, reducing the colinear part to 60%. The sequence homologies were highest between CCV and TGEV on the one hand and between FECV and FIPV on the other. The expression product of the CCV and the FECV ORF 6b can be detected in infected cells by immunoprecipitation.


title: Dynamic changes in blood cytokine levels as clinical indicators in severe acute respiratory syndrome.

text: OBJECTIVE To investigate the dynamic changes observed in serum levels of interleukins (ILs), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) in severe acute respiratory syndrome (SARS) patients. METHODS Sixty-one cases of SARS with positive antibodies to SARS coronavirus (SARS-CoV) were classified into the following categories: initial stage (3 - 7 days), peak stage (8 - 14 days), and remission and recovery stage (15 - 27 days). Forty-four healthy individuals were used as controls. Serum levels of ILs, TNF-alpha and TGF-beta 1 were measured in all subjects. Serum antibodies to SARS-CoV were detected only in SARS cases. RESULTS The mean concentration of serum IL-6 in SARS patients did not differ from that in the control group in initial and peak stages, but became significantly higher in remission and recovery stage compared with the control group, initial and peak stages (P < 0.01). The mean concentration of serum IL-8 in SARS patients did not differ from that of the control group in initial stage, but was significantly higher than control group in peak stage and remission and recovery stage (P < 0.05). And it was more significantly higher in remission and recovery stage than in peak stage (P < 0.01). The mean concentrations of IL-16 and TNF-alpha in SARS patients were higher than those of the control group for every length of the clinical courses investigated, and were especially high in remission and recovery stage (P < 0.01). SARS patients experienced higher concentration of serum IL-13 compared with the controls in initial stage (P < 0.01), but returned to normal levels in peak stage and in remission and recovery stage. The mean concentration of serum IL-18 in SARS patients was significantly lower than that of the control group during all clinical courses (P < 0.05). The mean concentration of serum TGF-beta1 in SARS patients was higher than that of the control group during all clinical courses. Although TGF-beta1 in serum decreased in remission and recovery stage in SARS patients, the average was still higher than that of the control group (P < 0.01). CONCLUSIONS Most proinflammatory cytokines and TGF-beta 1 were elevated during the early phase of SARS, which may be associated with lung infiltration and proliferation. Concurrently, the mean concentration of serum IL-13 decreased gradually, and the mean concentration of serum IL-18 level in SARS patients was lower than that of the control group during all the courses of SARS, suggesting that the immune state of the patients with SARS was obviously abnormal. Observing the dynamic changes in blood cytokine levels can provide a scientific basis to assess pathogenesis and efficacy of clinical treatment of SARS.


title: Membrane assembly of the triple-spanning coronavirus M protein. Individual transmembrane domains show preferred orientation.

text: The M protein of mouse hepatitis virus strain A59 is a triple-spanning membrane protein which assembles with an uncleaved internal signal sequence, adopting an NexoCcyt orientation. To study the insertion mechanism of this protein, domains potentially involved in topogenesis were deleted and the effects analyzed in topogenesis were deleted and the effects analyzed in several ways. Mutant proteins were synthesized in a cell-free translation system in the presence of microsomal membranes, and their integration and topology were determined by alkaline extraction and by protease-protection experiments. By expression in COS-1 and Madin-Darby canine kidney-II cells, the topology of the mutant proteins was also analyzed in vivo. Glycosylation was used as a biochemical marker to assess the disposition of the NH2 terminus. An indirect immunofluorescence assay on semi-intact Madin-Darby canine kidney-II cells using domain-specific antibodies served to identify the cytoplasmically exposed domains. The results show that each membrane-spanning domain acts independently as an insertion and anchor signal and adopts an intrinsic preferred orientation in the lipid bilayer which corresponds to the disposition of the transmembrane domain in the wild-type assembled protein. These observations provide further insight into the mechanism of membrane integration of multispanning proteins. A model for the insertion of the coronavirus M protein is proposed.


Reflections on 45 years as a clinical hematologist.


title: Vaccinia as a vector for gene delivery.

text: Gene therapy is a promising approach, yet so far it has shown limited effectiveness in many clinical trials, mainly due to insufficient gene transduction. Recombinant vaccinia virus (rVV) has been well developed as a gene delivery vector, initially for protein expression in mammalian cells. rVV has been further developed to express antigens in vivo in generating immunity for protection against specific infectious diseases and cancer. rVVs, as non-replicating viral vectors, have been demonstrated for their great potential as vaccines, for their diminished cytopathic effects, high levels of protein expression and strong immunogenicity, and they are relatively safe in animals and in human patients. A number of clinical trials using rVVs as vaccines have shown promising results for treating infectious diseases and cancer. In the last few years, due to its exceptional ability to replicate in tumour cells, the Western Reserve strain vaccinia has been explored as a replicating oncolytic virus for cancer virotherapy. As more is learned about the functions of viral gene products in controlling the mammalian cell cycle and in disabling cellular defence mechanisms, specific viral functions can be augmented or eliminated to enhance antitumour efficacy and improve tumour cell targeting. General mechanisms by which this oncolytic virus achieves the antitumour efficacy and specificity are reviewed. Specifically, the deletion of the viral genes for thymidine kinase and vaccinia growth factor resulted in a vaccinia mutant with enhanced tumour targeting activity and fully retaining its efficiency of replication in cancer cells. Other potential strategies for improving this vector for gene delivery will also be discussed in this review.


title: Quantification and characterization of pleural fluid in healthy dogs with thoracostomy tubes.

text: OBJECTIVE To quantify and characterize pleural fluid collected from healthy dogs after placement of a thoracostomy tube (TT). ANIMALS 8 healthy Coonhound-cross dogs (mean ± SD weight, 27.2 ± 1.6 kg). PROCEDURES Thoracic CT of each dog was performed before placement of a TT and daily thereafter for 7 days. Thoracic fluid volume was calculated from CT images. Effusion was aspirated when detected; volume was recorded, and cytologic analysis and bacterial culture were performed. RESULTS Mean ± SD volume of pleural effusion detected by CT was 1.43 ± 0.59 mL/kg (range, 0.12 to 3.32 mL/kg). Mean volume collected via aspiration was 0.48 ± 0.84 mL/kg (range, 0 to 2.16 mL/kg). Cytologic analysis yielded results consistent with an exudate, characterized by septic suppurative inflammation in 6 dogs and mixed inflammation in 1 dog; there was insufficient volume for analysis in 1 dog. Sufficient volume was obtained for bacterial culture for 6 dogs, which yielded pure growths of Staphylococcus pseudintermedius (n = 3) and Streptococcus equi subspecies zooepidemicus (2) and mixed growth of both of these species (1). The TT was removed before day 7 in 4 dogs because of pyothorax (n = 3) and irreversible damage to the TT (1). CONCLUSIONS AND CLINICAL RELEVANCE Presence of a TT induced a minimal volume of pleural effusion in healthy dogs. Pyothorax developed in most dogs between 4 and 6 days after TT placement. On the basis of these findings, a TT should be removed by the fourth day after placement, unless complications are detected sooner.


docid: tdzxgklk

title: Fluid administration, vasopressor use and patient outcomes in a group of high-risk cardiac surgical patients receiving postoperative goal-directed haemodynamic therapy: a pilot study.

text: The role of goal-directed therapy in high-risk cardiac surgical patients has not been determined. This study sought to observe the effect of a postoperative standardised haemodynamic protocol (SHP) on the administration of fluid and vasoactive drugs after high-risk cardiac surgery. This was an interventional pilot study. In 2010 to 2011, the SHP was introduced to the ICU at Wellington Regional Hospital, Wellington, New Zealand, for the perioperative management of patients undergoing high-risk cardiac surgery. A pulmonary artery catheter was inserted in the patients in the study group and fluids and supportive medications were provided in the ICU according to a protocol that targeted a cardiac index ≥ 2 l/min/m², mixed venous oxygen saturation ≥ 60% and a mean arterial pressure of 65 to 75 mmHg. Data from 40 consecutive high-risk cardiac surgical patients assigned to this protocol were compared with a matched cohort of 40 consecutive high-risk cardiac surgical patients receiving 'usual care' in 2009. Baseline characteristics were similar in the two groups. There was no significant difference in the duration of noradrenaline infusion in the SHP cohort compared to historical controls (median [IQR] 18.5 hours [31.63] versus 18 hours [18.3]; P=0.35), despite patients receiving more fluid in their first 12 hours in the ICU (mean 4687 ml [SD ± 2284 ml] versus 1889 ml [SD ± 1344 ml]; P <0.001). The SHP cohort had a higher rate of reintubation (4 in 37 [10.8%] versus 0 in 40 [0%]; P=0.049). The SHP delivered significantly more fluid, but did not reduce the duration of noradrenaline infusion, compared to usual care.


title: [Some medical staff positive for serum SARS coronavirus antibody IgG have only mild symptoms].

text: OBJECTIVE To identify patients with SARS coronavirus infection who have only mild symptoms. METHOD Enzyme-linked immunosorbent assay was employed to detect serum antibody against SARS coronavirus in the lysate of whole SARS coronavirus from 19 SARS patients and 200 medical staff members without obvious SARS symptoms after possible exposure to the virus during routine medical practice. RESULTS Serum IgG antibody against SARS coronavirus was detected in all the 19 SARS patients, and among the 200 staff members, 20 (10%) were found positive for the antibody but with no obvious or only mild symptoms. CONCLUSION Serum IgG antibody against SARS coronavirus is positive in a small proportion (around 10%) of the medical staff members exposed to the virus in our hospital, but may not cause obvious symptoms, suggesting SARS coronavirus infection might in some cases have mild or even no clinical manifestations.


title: Hepatobiliary pathology.

text: PURPOSE OF REVIEW Recent papers on disorders of the liver and biliary tract which clarify their pathogenesis and attendant morphologic changes are highlighted. RECENT FINDINGS The concept of 'bystander hepatitis' was cited in studies showing hepatic infiltration of CD8-positive T cells in the setting of extrahepatic infections such as influenza virus and severe acute respiratory syndrome. Diabetic liver lesions include glycogenic hepatopathy (in which poor diabetic control leads to swollen, glycogen-filled hepatocytes without fat, steatohepatitis or fibrosis) and diabetic hepatosclerosis in which there is diffuse perisinusoidal fibrosis (type IV collagen) without zonal predilection. Ground-glass hepatocellular inclusions (positive with periodic acid-Schiff stain for glycogen) were reported in three separate series of patients who were hepatitis B virus-negative, often transplant recipients, immunosuppressed and on multiple medications. A Banff consensus paper expertly compared and contrasted the histologic features which characterize the various causes of late liver allograft dysfunction. SUMMARY Informative papers emerged this past year concerning collateral damage to the liver in extrahepatic infections, diabetic lesions and causes of liver dysfunction after transplantation, among other topics.


title: Anosmia: diagnosis and management.

text: Disorders of the sense of smell can be frustrating for both the patient and physician. Ongoing research in this field has provided insight into the possible mechanisms for smell loss; however, therapy is still limited. Commercially distributed smell testing kits and newer screening tests using material available in all clinical settings have made diagnosis and measurement of the degree of impairment available to all physicians. A detailed history and physical examination are the most powerful tools in the evaluation of smell disorders, whereas imaging studies are reserved for preoperative planning or detailed assessment of positive physical findings.


title: Clinical aspects of immunosuppression in poultry.

text: Chickens, turkeys, and other poultry in a production environment can be exposed to stressors and infectious diseases that impair innate and acquired immunity, erode general health and welfare, and diminish genetic and nutritional potential for efficient production. Innate immunity can be affected by stressful physiologic events related to hatching and to environmental factors during the first week of life. Exposure to environmental ammonia, foodborne mycotoxins, and suboptimal nutrition can diminish innate immunity. Infectious bursal disease (IBD), chicken infectious anemia (CIA), and Marek's disease (MD) are major infectious diseases that increase susceptibility to viral, bacterial, and parasitic diseases and interfere with acquired vaccinal immunity. A shared feature is lymphocytolytic infection capable of suppressing both humoral and cell-mediated immune functions. Enteric viral infections can be accompanied by atrophic and depleted lymphoid organs, but the immunosuppressive features are modestly characterized. Some reoviruses cause atrophy of lymphoid organs and replicate in blood monocytes. Enteric parvoviruses of chickens and turkeys merit further study for immunosuppression. Hemorrhagic enteritis of turkeys has immunosuppressive features similar to IBD. Other virulent fowl adenoviruses have immunosuppressive capabilities. Newcastle disease can damage lymphoid tissues and macrophages. Avian pneumovirus infections impair the mucociliary functions of the upper respiratory tract and augment deeper bacterial infections. Recognition of immunosuppression involves detection of specific diseases using diagnostic tests such as serology, etiologic agent detection, and pathology. Broader measurements of immunosuppression by combined noninfectious and infectious causes have not found general application. Microarray technology to detect genetic expression of immunologic mediators and receptors offers potential advances but is currently at the developmental state. Control methods for immunosuppressive diseases rely largely on minimizing stress, reducing exposure to infectious agents through biosecurity, and increasing host resistance to infectious immunosuppressive diseases by vaccination. A longer-term approach involves genetic selection for resistance to immunosuppressive diseases, which has shown promising results for MD but equivocal results for IBD and CIA.


Role for primary care in epidemic surge capacity.


Application of non-invasive ventilation in China over 20 years.


title: The 3a accessory protein of SARS coronavirus specifically interacts with the 5'UTR of its genomic RNA, Using a unique 75 amino acid interaction domain.

text: More than four years have passed since the outbreak of the severe acute respiratory syndrome (SARS) epidemic, and still very little is known about the molecular biology and pathogenesis of this deadly virus. Among the accessory proteins of the SARS coronavirus (SARS-CoV), the 3a protein has been shown to interact with the spike, envelope, and membrane glycoprotein and has recently been established to be a structural component of capsid. Recent studies suggest that the 3a protein may function as an ion channel and may promote virus release. In order to further characterize the functional properties of this protein, we initiated studies to check its RNA binding activity. Using the yeast three-hybrid system, electrophoretic mobility shift assay (EMSA), and ultraviolet (UV) cross-linking techniques, we have shown that the 3a protein is capable of binding specifically to the 5' untranslated region (5'UTR) of the SARS virus genomic RNA. Further, we have mapped the interaction domain of the 3a protein responsible for this RNA-protein interaction using a series of deletion mutants and defined it to the central 75 amino acid region. This RNA binding motif of 3a does not share homology with any other known RNA binding protein and may have an important role in viral capsid assembly and pathogenesis.


title: Should patients stop their radiotherapy or chemotherapy during the COVID-19 pandemic.

text: In the previous stage, there were too many patients with Corona virus disease 2019 (COVID-19) in Wuhan. Ordinary people, patients, even doctors, had a great sense of desperate. On the one hand, almost all doctors participated in the treatment of patients of COVID-19. On the other hand, the government restricted residents to go out, and the sick people were also afraid of being infected with COVID-19 when seeking medical treatment. Whether cancer patients seek medical treatment or not has become a contradiction for a long time. Our Viewpoint paper is to provide a positive signal to doctors and patients that patients with in the middle or advanced stage of cancer can receive radiotherapy and/or chemotherapy normally under protective measures.


title: The impact of the SARS epidemic on the utilization of medical services: SARS and the fear of SARS.

text: Using interrupted time-series analysis and National Health Insurance data between January 2000 and August 2003, this study assessed the impacts of the severe acute respiratory syndrome (SARS) epidemic on medical service utilization in Taiwan. At the peak of the SARS epidemic, significant reductions in ambulatory care (23.9%), inpatient care (35.2%), and dental care (16.7%) were observed. People's fears of SARS appear to have had strong impacts on access to care. Adverse health outcomes resulting from accessibility barriers posed by the fear of SARS should not be overlooked.


Cryptosporidiosis and proliferative ileitis in a hamster.


title: Inhibitory effects of recombinant human cystatin C on human coronaviruses.

text: Cystatin C, a potent inhibitor of cysteine proteases such as papain and cathepsin B, was examined for its effect on human coronaviruses OC43 and 229e. Both viruses were greater than 99% inhibited by 0.1 mM inhibitor. Endpoint titrations showed that inhibiting activity paralleled that of leupeptin, a serine and cysteine protease inhibitor, and indicated that 1 to 2 microM inhibitor, slightly above physiologic levels, was effective.


Q & A: Bob Michell.


docid: xvijxsrn

title: Viral antibody studies of laboratory dogs with diarrheal disease.

text: Viral antibody studies were done on laboratory dogs in an epizootic of gastrointestinal disease. Increased hemagglutination-inhibition antibody titers to a parvovirus (PV) antigenically related to feline panleukopenia virus were found in convalescent serum specimens of 78% (20/26) of the affected dogs and in 83% (5/6) of apparently healthy dogs. With one exception, all dogs tested had significant levels of hemagglutination-inhibition antibody to this PV. Similar increased antibody titers were found to feline panleukopenia virus. Also, neutralizing antibody responses were detected to the canine coronavirus in 24% (6/25) and canine herpesvirus in 45% (10/22) of the affected dogs. However, antibody titers did not increase to canine distemper virus, infectious canine hepatitis virus, canine parainfluenza virus, or minute virus of canines. Subsequent serotesting of the colony provided evidence that additional PV infections occurred in pups from each of 8 litters born 3 to 8 months after the epizootic. These findings indicated the continued presence of the PV for more than 1 year in the infected colony. Of 19 laboratory personnel who worked with the affected dogs, none, including 4 with a concurrent diarrheal disease, developed or had antibodies to the PV or canine coronavirus.


title: Endovascular stent placement for chronic post-thrombotic symptomatic ilio-femoral venous obstructive lesions: a single-center study of safety, efficacy and quality-of-life improvement.

text: BACKGROUND Post-thrombotic syndrome (PTS) is a frequent complication of deep vein thrombosis (DVT) despite adequate treatment. Venous angioplasty and stent placement has been progressively used to restore and maintain venous patency in PTS patients. This study reports our single-center experience with the use of endovascular treatment for chronic post-thrombotic symptomatic ilio-femoral venous obstructive lesions. METHODS A prospective mono-centric observational cohort study of PTS patients with chronic symptomatic ilio-femoral venous obstructive lesions referred for endovascular treatment was conducted from March 2012 to April 2016. Procedure consisted in recanalization, pre-dilation and self-expandable stenting of stenotic or occluded iliac and/or femoral veins. Severity of PTS, quality-of-life and treatment outcomes were assessed using Villalta scale and Chronic Venous Insufficiency Questionnaire (CIVIQ-20) at baseline and 3 months after the procedure. Imaging follow-up was based on duplex ultrasound (US) and computed tomography (CT). RESULTS Twenty-one patients (11 females, 10 males; median age, 41 years; range, 32-60) were included. Recanalization and stenting was successfully accomplished in all prime procedures, 4 patients benefitted from an additional procedure. Immediate technical success rate was 96% considering 25 procedures, performed without any complications. Median follow-up was 18 months (range, 6-30 months) with a 90.5% stent patency rate. Villalta score significantly decreased from baseline compared with 3 months after the procedure [14 (range, 11-22) and 5 (range, 1-10), respectively, P<0.0001], showing a significant decrease in the severity of PTS. CIVIQ-20 score significantly decreased from baseline compared with 3 months after stenting [48.5 (range, 39-73) and 26.5 (range, 21-45), respectively, P<0.0001] thus showing a significant improvement of quality-of-life. Post-procedural CIVIQ-20 score was significantly associated with Villalta score (95% CI, 1.53-2.95; P<0.0001). CONCLUSIONS Our results confirm the high clinical success rate and safety of endovascular PTS treatment and highlight the significant impact of stenting on the quality of life of patients with chronic symptomatic ilio-femoral venous obstructive lesions.


title: Metabolic and Hormonal Determinants of Glomerular Filtration Rate and Renal Hemodynamics in Severely Obese Individuals.

text: OBJECTIVE Renal function is often compromised in severe obesity. A true measurement of glomerular filtration rate (GFR) is unusual, and how estimation formulae (EstForm) perform in such individuals is unclear. We characterized renal function and hemodynamics in severely obese individuals, assessing the reliability of EstForm. METHODS We measured GFR (mGFR) by iohexol plasma clearance, renal plasma flow (RPF) by 123I-ortho-iodo-hippurate, basal and stimulated vascular renal indices, endothelium-dependent and -independent vasodilation using flow-mediated dilation (FMD) as well as metabolic and hormonal profile in morbid, otherwise healthy, obese subjects. RESULTS Compared with mGFR, the better performing EstForm was CKD-EPI (5.3 ml/min/1.73 m2 bias by Bland-Altman analysis). mGFR was directly related with RPF, total and incremental glucose AUC, and inversely with PTH and h8 cortisol. Patients with mGFR below the median shown significantly higher PTH and lower vitamin D3. Basal or dynamic renal resistive index, FMD, pulse wave velocity were not related with mGFR. In an adjusted regression model, renal diameter and plasma flow remained related with mGFR (R2 = 0.67), accounting for 15% and 21% of mGFR variance, respectively. CONCLUSIONS CKD-EPI formula should be preferred in morbid obesity; glucose increments during oral glucose tolerance test correlate with hyperfiltration; RPF and diameter are independent determinants of mGFR; slightly high PTH values, frequent in obesity, might influence mGFR.


title: Progressive and nonresolving pneumonia.

text: PURPOSE OF REVIEW Patients with progressive and/or nonresolving community-acquired pneumonia are at risk for increased morbidity and mortality. It is critical to be able to identify patients at risk to institute early appropriate therapy. The purpose of this review is to summarise the most updated developments in this area. RECENT FINDINGS This review will glean from the recent literature clinical, laboratory, and radiologic findings that help identify patients at risk for such complications of their pneumonia. New studies will be reviewed that have identified some of the causes for treatment failures including the type of pathogen and discordant antimicrobial therapy. It will also discuss newly recognised and emerging infectious diseases that may result in progressive or nonresponding pneumonia including severe acute respiratory syndrome, avian influenzae, severe group A streptococcal disease, and community-acquired methicillin-resistant Staphylococcus aureus. Promising treatments have been identified for patients with progressive pneumonia due to an overwhelming host immune response including activated protein C and intravenous immunoglobulin. SUMMARY Both progressive and nonresolving pneumonia represent treatment failure as a result of inappropriate initial therapy, a noninfectious cause, or an overwhelming immune response. It is critical to be able to identify patients with nonresponding pneumonia and to identify patients at risk for progressive pneumonia to institute appropriate therapy.


Influenza in the Eastern Mediterranean Region: identifying the unknowns for detection and control of epidemic and pandemic threats.


title: Intracellular restriction of a productive noncytopathic coronavirus infection.

text: Virus infection in vitro can either result in a cytopathic effect (CPE) or proceed without visible changes in infected cells (noncytopathic infection). We are interested in understanding the mechanisms controlling the impact of coronavirus infection on host cells. To this end, we compared a productive, noncytopathic infection of murine hepatitis virus (MHV) strain A59 in the fibroblastlike cell line NIH 3T3 with cytopathic MHV infections. Infected NIH 3T3 cells could be cultured for up to 4 weeks without apparent CPE and yet produce virus at 10(7) to 10(8) PFU/ml. Using flow cytometry, we demonstrated that NIH 3T3 cells expressed as much MHV receptor CEACAM1 as other cell lines which die from MHV infection. In contrast, using quantitative reverse transcription-PCR and metabolic labeling of RNA, we found that the rate of viral RNA amplification in NIH 3T3 cells was lower than the rate in cells in which MHV induces a CPE. The rate of cellular RNA synthesis in contact-inhibited confluent NIH 3T3 cells was also lower than in cells permissive to cytopathic MHV infection. However, the induction of cellular RNA synthesis in growing NIH 3T3 cells did not result in an increase of either viral RNA amplification or CPE. Our results suggest that a specific, receptor CEACAM1-independent mechanism restricting coronaviral RNA synthesis and CPE is present in NIH 3T3 and, possibly, other cells with preserved contact inhibition.


title: Comparison of the replication of influenza A viruses in Chinese ring-necked pheasants and chukar partridges.

text: We investigated the replication and transmission of avian influenza A viruses in two species thought to be intermediate hosts in the spread of influenza A viruses in live poultry markets: Chinese ring-necked pheasants and chukar partridges. All 15 hemagglutinin subtypes replicated in pheasants, and most subtypes transmitted to naïve contact pheasants, primarily via the fecal-oral route. Many viruses were shed from the gastrointestinal tract of experimentally inoculated pheasants for 14 days or longer. Virus was isolated from the cloacal swabs of one contact pheasant for an unprecedented 45 days. Chukar partridges were less susceptible to infection with avian influenza viruses. The viruses that replicated in chukar partridges were isolated for 7 days after experimental inoculation, predominantly from the respiratory tract. We detected high neutralizing antibody titers with correspondingly low levels of serum hemagglutination inhibition antibody titers in pheasants and chukar partridges when chicken red blood cells were used in serological analyses. When horse erythrocytes were used, antibody titers were comparable to those obtained by using the neutralization assay. More importantly, the results suggested that pheasants can serve as a reservoir of influenza virus. Because of their continuous asymptomatic infection and longer stay in the markets, pheasants are ideal "carriers" of influenza A viruses. Their continued presence in live markets contributes to the perpetuation and genetic interaction of influenza viruses there. On the basis of our findings, it does not make good sense to ban quail but not pheasants from the live markets.


title: Investigation of an outbreak of acute respiratory disease in côte d'ivoire in april 2007.

text: BACKGROUND This study aim was to investigate an outbreak of human cases of unexplained influenza-like illness and fatal acute respiratory infection (ARI), with simultaneous poultry illness and high mortality raising concerns of possible influenza A (H5N1), virus in Cote d'Ivoire in February and March 2007. MATERIALS AND METHODS To investigate the outbreak, we conducted active surveillance in the community and reviewed health registries. Persons meeting the case definition were asked to provide nasopharyngeal specimens. On the basis of clinical and epidemiological information, specimens were tested using conventional RT-PCR for the M gene of the influenza viruses and hemagglutinin H5 of avian influenza A (H5N1), virus; negative samples were tested for other respiratory viruses. Specimens from healthy animals were also collected. RESULTS Between October 2006, and February 2007, 104 suspected cases of Acute Respiratory Disease that included; 31 deaths recorded. We collected and tested 73 nasopharyngeal specimens; of which, 2, were positive for human Coronavirus OC43 and 1 for influenza C virus. No pathogens were identified in animal specimens. CONCLUSIONS The investigation quickly ruled out influenza A (H5N1), virus as the cause and found laboratory-confirmed cases of influenza C virus and human Coronavirus OC 43 for the first time in both Côte d'Ivoire and in a Sub-Saharan African country. However we were not able to show that these viruses caused the outbreak. Monitoring of influenza viruses must be a priority but other respiratory viruses and non-viral causes may be of interest too.


Combined laparoscopic excisional surgery for synchronous endometrial and rectal adenocarcinoma in an obese woman.


docid: crvehfnr

title: The distribution of human coronavirus strain 229E on the surface of human diploid cells.

text: The distribution of human coronavirus strain 229E (HCV 229E) particles on the surface of human diploid (MRCc) cells was examined. Virus particles showed a totally random distribution on fixed cells and on cells to which virus had been adsorbed in the cold. A marked redistribution of virus particles was observed on warming virus-cell preparations to 33 degrees C for 20 min, the peripheral areas of the cell becoming relatively devoid of virus particles while the majority of particles were now located some distance from the edge of the cell. Redistribution did not occur in the presence of metabolic inhibitors.


title: A positive-strand RNA virus with three very different subgenomic RNA promoters.

text: Numerous RNA viruses generate subgenomic mRNAs (sgRNAs) for expression of their 3'-proximal genes. A major step in control of viral gene expression is the regulation of sgRNA synthesis by specific promoter elements. We used barley yellow dwarf virus (BYDV) as a model system to study transcriptional control in a virus with multiple sgRNAs. BYDV generates three sgRNAs during infection. The sgRNA1 promoter has been mapped previously to a 98-nucleotide (nt) region which forms two stem-loop structures. It was determined that sgRNA1 is not required for BYDV RNA replication in oat protoplasts. In this study, we show that neither sgRNA2 nor sgRNA3 is required for BYDV RNA replication. The promoters for sgRNA2 and sgRNA3 synthesis were mapped by using deletion mutagenesis. The minimal sgRNA2 promoter is approximately 143 nt long (nt 4810 to 4952) and is located immediately downstream of the putative sgRNA2 start site (nt 4809). The minimal sgRNA3 core promoter is 44 nt long (nt 5345 to 5388), with most of the sequence located downstream of sgRNA3 start site (nt 5348). For both promoters, additional sequences upstream of the start site enhanced sgRNA promoter activity. These promoters contrast to the sgRNA1 promoter, in which almost all of the promoter is located upstream of the transcription initiation site. Comparison of RNA sequences and computer-predicted secondary structures revealed little or no homology between the three sgRNA promoter elements. Thus, a small RNA virus with multiple sgRNAs can have very different subgenomic promoters, which implies a complex system for promoter recognition and regulation of subgenomic RNA synthesis.


title: An exposed domain in the severe acute respiratory syndrome coronavirus spike protein induces neutralizing antibodies.

text: Exposed epitopes of the spike protein may be recognized by neutralizing antibodies against severe acute respiratory syndrome (SARS) coronavirus (CoV). A protein fragment (S-II) containing predicted epitopes of the spike protein was expressed in Escherichia coli. The properly refolded protein fragment specifically bound to the surface of Vero cells. Monoclonal antibodies raised against this fragment recognized the native spike protein of SARS CoV in both monomeric and trimeric forms. These monoclonal antibodies were capable of blocking S-II attachment to Vero cells and exhibited in vitro antiviral activity. These neutralizing antibodies mapped to epitopes in two peptides, each comprising 20 amino acids. Thus, this region of the spike protein might be a target for generation of therapeutic neutralizing antibodies against SARS CoV and for vaccine development to elicit protective humoral immunity.


title: Identification of linear heparin-binding peptides derived from human respiratory syncytial virus fusion glycoprotein that inhibit infectivity.

text: It has been shown previously that the fusion glycoprotein of human respiratory syncytial virus (RSV-F) interacts with cellular heparan sulfate. Synthetic overlapping peptides derived from the F-protein sequence of RSV subtype A (strain A2) were tested for their ability to bind heparin using heparin-agarose affinity chromatography (HAAC). This evaluation identified 15 peptides representing eight linear heparin-binding domains (HBDs) located within F1 and F2 and spanning the protease cleavage activation site. All peptides bound to Vero and A549 cells, and binding was inhibited by soluble heparins and diminished by either enzymatic treatment to remove cell surface glycosaminoglycans or by treatment with sodium chlorate to decrease cellular sulfation. RSV-F HBD peptides were less likely to bind to glycosaminoglycan-deficient CHO-745 cells than parental CHO-K1 cells that express these molecules. Three RSV-F HBD peptides (F16, F26, and F55) inhibited virus infectivity; two of these peptides (F16 and F55) inhibited binding of virus to Vero cells, while the third (F26) did not. These studies provided evidence that two of the linear HBDs mapped by peptides F16 and F55 may mediate one of the first steps in the attachment of virus to cells while the third, F26, inhibited infectivity at a postattachment step, suggesting that interactions with cell surface glycosaminoglycans may play a role in infectivity of some RSV strains.


title: Characterization of murine coronavirus RNA by hybridization with virus-specific cDNA probes.

text: Genome RNA of mouse hepatitis virus (MHV) strain A59 has been used as a template to synthesize two virus-specific probes: cDNArep, representing the majority of sequences of the genome RNA and cDNA3', representing the 3' end of the genome RNA. Molecular hybridization with these cDNAs was used to characterize both genome RNA and intracellular virus-specific RNAs. Hybridization of genome RNAs of MHV strains A59, JHM, and MHV-3 with A59 cDNArep showed that, although these three strains exhibit different pathogenicities, they contain closely related nucleotide sequences. Hybridization of intracellular RNA from MHV-infected cells with virus-specific cDNA shows that (i) the majority of virus-specific RNA is polyadenylated, (ii) virus-specific intracellular RNA contains six subgenomic species of the same polarity as genome RNA and (iii) all subgenomic RNAs contain the same 3' sequences as the genome RNA and thus form a nested set of RNAs.


Detection of body temperature with infrared thermography: accuracy in detection of fever.


The open university.


Characterization of a small plaque mutant of the A59 strain of mouse hepatitis virus defective in cell fusion.


title: The treatment of hospital wastewater: an appraisal.

text: Hospitals discharge considerable amounts of chemicals and microbial agents in their wastewaters. Problem chemicals present in hospital wastewater belong to different groups, such as antibiotics, X-ray contrast agents, disinfectants and pharmaceuticals. Many of these chemical compounds resist normal wastewater treatment. They end up in surface waters where they can influence the aquatic ecosystem and interfere with the food chain. Humans are particularly exposed by the drinking water, produced from surface water. Microbial agents of special concern are multiresistant microbial strains. The latter are suspected to contribute to the spread of antibiotic resistance. In this paper, we will discuss the different approaches towards hospital wastewater treatment. The principle of uncoupling hospitals from public sewers warrants indepth evaluation by technologists and ecotoxicologists as well as public health specialists.


title: Japanese Encephalitis Vaccines.

text: Japanese encephalitis (JE) is a significant human health concern in Asia, Indonesia and parts of Australia with more than 3 billion people potentially at risk of infection with Japanese encephalitis virus (JEV), the causative agent of JE. Given the risk to human health and the theoretical potential for JEV use as a bioweapon, the development of safe and effective vaccines to prevent JEV infection is vital for preserving human health. The development of vaccines for JE began in the 1940s with formalin-inactivated mouse brain-derived vaccines. These vaccines have been shown to induce a protective immune response and to be very effective. Mouse brain-derived vaccines were still in use until May 2011 when the last lots of the BIKEN(®) JE-VAX(®) expired. Development of modern JE vaccines utilizes cell culture-derived viruses and improvements in manufacturing processes as well as removal of potential allergens or toxins have significantly improved vaccine safety. China has developed a live-attenuated vaccine that has proven to induce protective immunity following a single inoculation. In addition, a chimeric vaccine virus incorporating the prM and E structural proteins derived from the live-attenuated JE vaccine into the live-attenuated yellow fever 17D vaccine virus backbone is currently in clinical trials. In this article, we provide a summary of JE vaccine development and on-going clinical trials. We also discuss the potential risk of JEV as a bioweapon with a focus on virus sustainability if used as a weapon.


title: Characterization of an antigenically distinct porcine rotavirus.

text: A porcine virus with rotavirus morphology, which was antigenically unrelated to previously described rotaviruses, is described. Particles with an outer capsid layer measured 75 nm and those lacking the outer layer were 63 nm in diameter. Particles which resembled cores were also identified. The virus was shown to be antigenically distinct from other rotaviruses as judged by immunofluorescence and immune electron microscopy, and it failed to protect piglets from challenge with porcine rotavirus. Analysis of the genome indicated that it was composed of 11 segments of double-stranded RNA with the same overall size range as other rotaviruses but with an unusual segment pattern.


title: Options in topical therapies in the management of patients with acute pain.

text: The traditional cornerstones of analgesic therapy for patients with acute pain have been oral therapies; however, all oral agents exhibit a variety of potentially dose-limiting or intolerable adverse effects in patients. Elderly patients and those with concomitant conditions already being managed with multiple systemic drugs may be particularly susceptible to systemic toxicities with oral analgesic therapies. Topical agents offer an alternative to oral modalities and can effectively treat patients with acute pain while offering lower systemic absorption and conferring little risk of systemic toxicity. The objective of this article is to review the therapeutic usefulness of available topical therapies in their most thoroughly investigated applications, the treatment of patients with acute musculoskeletal and herpetic pain. For example, although heating pads/wraps and cold packs are widely used to alleviate pain associated with sprains, strains, and contusions, evidence of the effectiveness of these methods is lacking. However, there are sufficient data supporting the use of various topical formulations of nonsteroidal anti-inflammatory drugs (NSAIDs) for these indications (ketoprofen gel or patch, ibuprofen gel or cream, and diclofenac gel or patch), and demonstrating markedly less patient risk of systemic toxicity than is associated with oral NSAID therapy. A ketoprofen patch was shown to be effective and well tolerated in the treatment of patients with tendinopathies. In the treatment of acute neck or low back pain, cold and heat therapies have demonstrated limited effectiveness for patients, and the efficacy of topical NSAIDs has not been established. Use of topical NSAID therapy has been useful in reducing acute-phase herpes zoster pain, and the lidocaine 5% patch has been shown to reduce acute herpetic pain intensity once lesions have healed (the patch cannot be applied to open skin lesions). Topical analgesics represent an alternative treatment modality for patients experiencing acute pain who cannot or choose not to take oral therapies.


title: Licking latency with licorice.

text: Numerous viruses cause latent infections in humans, and reactivation often results in pain and suffering. While vaccines for several of these viruses are available or currently being studied in clinical trials, and antiviral therapies have been successful in preventing or treating active infection, therapy to eradicate latent infection has lagged behind. A new study reported in this issue of the JCI shows that treatment of cells latently infected with Kaposi sarcoma-associated herpesvirus (KSHV) with glycyrrhizic acid, a component of licorice, reduces synthesis of a viral latency protein and induces apoptosis of infected cells. This finding suggests a novel way to interrupt latency.


Detection of feline infectious peritonitis virus-like antigen in ferrets.


title: [Virological and serological characteristics of outbreaks and sporadic cases of acute gastroenteritis].

text: Specimens from patients with gastroenteritis (GE) collected during outbreaks and from sporadic cases reported in the USSR in 1979-1984 were examined by electron microscopy (EM), enzyme immunoassay, rotavirus neutralization test in cell culture. All the winter-spring outbreaks and a considerable number (34.9%) of sporadic GE cases were caused by rotaviruses. The summer-autumn outbreaks were of non-rotavirus nature. In water-borne winter-spring outbreaks in adults, severe forms of GE with signs of dehydration were observed. Among infants, cases of virus-carrier state were detected. The rate of rotavirus detection by EM in winter-spring outbreaks depended on the time of specimen collection and decreased after 4 days from the onset of the disease. Apart from rotaviruses, adeno-, astro-, calici-, coronaviruses, and picornavirus-like particles were detected by EM in feces from GE patients.


title: Detection and molecular characterization of gene 3 and 5 of turkey coronavirus from turkeys with severe enteritis in Brazil.

text: Turkey coronavirus (TCoV) is a causative agent associated with poult enteritis and mortality syndrome (PEMS) in turkeys worldwide. The disease is an acute, highly contagious enteric disease that is characterized by depression, anorexia, diarrhea, and high mortality in commercial turkey flocks. The presence of TCoV in 12 intestinal-content samples, from turkey flocks aged between 10 and 104 days and exhibiting severe enteritis, was monitored during the period of 2004 to 2006. TCoV detection was accomplished by a reverse transcriptase-polymerase chain reaction (RT-PCR) through amplification of the 3' UTR region, followed by amplification of genes 3 and 5. Molecular characterization of the viruses was done through amplification of genes 3 and 5 and showed evidence of genetic similarity between them, although they differed from sequences of other TCoVs described in the literature. In relation to gene 3, samples showed a greater relationship with chicken infectious bronchitis virus (IBV), while gene 5 showed greater identity with pheasant coronavirus (PhCoV). Our results suggest that the strategy of amplification of the 3' UTR region, followed by sequencing of genes 3 and 5, has proven to be an effective means of detecting TCoV in intestinal contents.


title: DNA accelerates the inhibition of human cathepsin V by serpins.

text: A balance between proteolytic activity and protease inhibition is crucial to the appropriate function of many biological processes. There is mounting evidence for the presence of both papain-like cysteine proteases and serpins with a corresponding inhibitory activity in the nucleus. Well characterized examples of cofactors fine tuning serpin activity in the extracellular milieu are known, but such modulation has not been studied for protease-serpin interactions within the cell. Accordingly, we present an investigation into the effect of a DNA-rich environment on the interaction between model serpins (MENT and SCCA-1), cysteine proteases (human cathepsin V and human cathepsin L), and cystatin A. DNA was indeed found to accelerate the rate at which MENT inhibited cathepsin V, a human orthologue of mammalian cathepsin L, up to 50-fold, but unexpectedly this effect was primarily effected via the protease and secondarily by the recruitment of the DNA as a "template" onto which cathepsin V and MENT are bound. Notably, the protease-mediated effect was found to correspond both with an altered substrate turnover and a conformational change within the protease. Consistent with this, cystatin inhibition, which relies on occlusion of the active site rather than the substrate-like behavior of serpins, was unaltered by DNA. This represents the first example of modulation of serpin inhibition of cysteine proteases by a co-factor and reveals a mechanism for differential regulation of cathepsin proteolytic activity in a DNA-rich environment.


title: Total sialic acid: an acute phase reactant in cats with a possible role in feline coronavirus infection.

text: The aims of this study were to validate a colorimetric method to measure total sialic acid (TSA) in feline serum and to investigate the serum concentration of TSA in clinically healthy cats seronegative (n = 9) and seropositive (n = 48) for feline coronavirus (FCoV), and in cats affected by feline infectious peritonitis (FIP, n = 28), tumors (n = 20), or inflammation (n = 16). The correlation between TSA and alpha(1)-acid glycoprotein (AGP) was also investigated. The method employed in this study is precise and accurate at TSA levels (in mg/L) commonly encountered in feline serum. No significant differences between seropositive (385.6 +/- 192.2 mg/L) and seronegative (433.5 +/- 179.0 mg/L) cats were detectable, suggesting that the simple infection by FCoVs does not influence TSA levels. Compared with seropositive controls, the concentration of TSA was higher in cats with FIP (556.7 +/- 268.3 mg/L, P = 0.003), tumors (522.5 +/- 294.4 mg/L, P = 0.028), and inflammation (546.8 +/- 208.3 mg/L, P = 0.018). The discriminating power of TSA for FIP is moderate (area under the ROC curve = 0.65) and the likelihood ratio is higher than 3.0 only at high TSA levels. Consequently, TSA could support a diagnosis of FIP only at extremely high serum concentration (> 800 mg/L) or when the pre-test probability of FIP is high. No correlations were found between the TSA and AGP concentrations in cats with FIP, suggesting that sialylated proteins other than AGP are present. Both the antibody titre and the degree of AGP sialylation were negatively correlated with TSA levels, suggesting that increased TSA may contribute to reduce the burden of FCoVs.


docid: ywaadybw

title: Novel double-stapling technique for distal oesophageal resection and oesophago-jejunal anastomosis.

text: AIM The restoration of the digestive tract by performing an esophago-jejunal anastomosis (EJA) is a crucial step of the total gastric and distal esophagus surgical resection for esophago-gastric junction (EGJ) cancer. We have already ideated and tested on a cadaver model an innovative technique which could be useful to minimize the risk of complications related to the phase of securing the anvil of the circular stapler prior to perform the EJA. This surgical technique was derived from the well-known "double-stapling Knight and Griffen" one that was described for the rectal resection. We used the following described technique in 20 patients with EGJ cancer and it is efficient, reliable, safe, easy to learn and easy to perform. MATERIALS AND METHODS From August 2014 to May 2015, 20 patients (14 male and 6 female) underwent surgery for esophagogastric junction cancer: In all patients a distal esophageal resection and total gastrectomy was performed. Through the trans-hiatal access, the free margins of the esophageal stump were suspended and the anvil of a circular stapler on a new dedicated and registered support bar was inserted into the lumen. Subsequently, the linear suturing stapler is closed over the bar and then fired to suture the distal stump of the esophagus; after the confirmation of a negative margin, the bar is retracted and the push-rod of the anvil is pulled out through the linear suture. Finally, the anastomosis is performed with the classic technique by using a circular stapler. RESULTS No postoperative mortality occurred; postoperative course has been uneventful for 18 patients. One patient developed anastomotic fistula that has been treated conservatively with endoscopic prothesis, removed after 20 days. One patient developed in 3 POD myocardial infarction Mean Hospital stay has been 14 days (range 7-20 days). CONCLUSIONS The aim of our new procedure is the insertion the anvil of a common circular stapler without handsewn securing; this is to reduce the technical difficulties related to the hand-sewn securing into a deep and narrow anatomic location, typical of the trans-hiatal approach. KEY WORDS Anastomosis, Oesophago-gastric junction cancer, Stapler, Trans-hiatal.


title: Novel beta-barrel fold in the nuclear magnetic resonance structure of the replicase nonstructural protein 1 from the severe acute respiratory syndrome coronavirus.

text: The nonstructural protein 1 (nsp1) of the severe acute respiratory syndrome coronavirus has 179 residues and is the N-terminal cleavage product of the viral replicase polyprotein that mediates RNA replication and processing. The specific function of nsp1 is not known. Here we report the nuclear magnetic resonance structure of the nsp1 segment from residue 13 to 128, which represents a novel alpha/beta-fold formed by a mixed parallel/antiparallel six-stranded beta-barrel, an alpha-helix covering one opening of the barrel, and a 3(10)-helix alongside the barrel. We further characterized the full-length 179-residue protein and show that the polypeptide segments of residues 1 to 12 and 129 to 179 are flexibly disordered. The structure is analyzed in a search for possible correlations with the recently reported activity of nsp1 in the degradation of mRNA.


title: Using protein homology models for structure-based studies: approaches to model refinement.

text: Homology modeling is a computational methodology to assign a 3-D structure to a target protein when experimental data are not available. The methodology uses another protein with a known structure that shares some sequence identity with the target as a template. The crudest approach is to thread the target protein backbone atoms over the backbone atoms of the template protein, but necessary refinement methods are needed to produce realistic models. In this mini-review anchored within the scope of drug design, we show the validity of using homology models of proteins in the discovery of binders for potential therapeutic targets. We also report several different approaches to homology model refinement, going from very simple to the most elaborate. Results show that refinement approaches are system dependent and that more elaborate methodologies do not always correlate with better performances from built homology models.


title: [Simultaneous detection of respiratory viruses and influenza A virus subtypes using multiplex PCR].

text: This study was conducted to investigate the respiratory viruses and subtyping of influenza A virus when positive by multiplex PCR in patients with flu-like symptoms, after the pandemic caused by influenza A (H1N1)pdm09. Nasopharyngeal swab samples collected from 700 patients (313 female, 387 male; age range: 24 days-94 yrs, median age: 1 yr) between December 2010 - January 2013 with flu-like symptoms including fever, headache, sore throat, rhinitis, cough, myalgia as defined by the World Health Organization were included in the study. Nucleic acid extractions (Viral DNA/RNA Extraction Kit, iNtRON, South Korea) and cDNA synthesis (RevertAid First Strand cDNA Synthesis Kits, Fermentas, USA) were performed according to the manufacturer's protocol. Multiplex amplification of nucleic acids was performed using DPO (dual priming oligonucleotide) primers and RV5 ACE Screening Kit (Seegene, South Korea) in terms of the presence of influenza A (INF-A) virus, influenza B (INF-B) virus, respiratory syncytial virus (RSV), and the other respiratory viruses. PCR products were detected by automated polyacrylamide gel electrophoresis using Screen Tape multiple detection system. Specimens which were positive for viral nucleic acids have been further studied by using specific DPO primers, FluA ACE Subtyping and RV15 Screening (Seegene, South Korea) kits. Four INF-A virus subtypes [human H1 (hH1), human H3 (hH3), swine H1 (sH1), avian H5 (aH5)] and 11 other respiratory viruses [Adenovirus, parainfluenza virus (PIV) types 1-4, human bocavirus (HBoV), human metapneumovirus (HMPV), rhinovirus types A and B, human coronaviruses (HCoV) OC43, 229E/NL63] were investigated with those tests. In the study, 53.6% (375/700) of the patients were found to be infected with at least one virus and multiple respiratory virus infections were detected in 15.7% (59/375) of the positive cases, which were mostly (49/59, 83%) in pediatric patients. RSV and rhinovirus coinfections were the most prevalent (18/29, 62.7%) dual infections. The distribution of 436 respiratory viruses identified from 375 patients were as follows; 189 (43.3%) RSV, 93 (21.4%) rhinovirus, 86 (19.8%) INF-A, seven (1.6%) INF-B, 22 (5%) PIV types 1-3, 14 (3.2%) HMPV, 11 (2.5%) HCoV, nine (2%) HBoV, and five (1.2%) adenovirus. Fifty-five (64%) out of 86 INF-A viruses were subtyped as hH3, 24 (27.9%) were sH1 and seven (8.1%) were hH1. Avian H5 was not detected in any samples. The overall prevalence rates of INF-A, INF-B, RSV and other respiratory viruses were 12%, 1%, 27%, and 14.6%, respectively. RSV was the most prevalent respiratory agent in pediatric (161/313, 51%) cases, while INF-A virus in adult (24/62, 38.7%) patients. Influenza viruses were detected as responsible pathogens in 13.3% (93/700) of the patients with flu-like symptoms. Among the cases, a 1-month-old baby was infected with three virus strains (INF-A hH1+INF-A sH1+HCoV OC43) and a 82-year-old patient was infected with two INF-A virus subtypes (hH3 + sH1). INF-A viruses were mostly detected (79/86) in winter period, from December to March. INF-A virus sH1, was the most prevalent subtype in flu cases till February 2011 (22/86), after replaced by INF-A virus hH3. Beginning from February 2012, a significant increase observed in the cases infected with INF-A virus subtype hH3 (39/86). In conclusion, the identification and surveillance of influenza virus types and subtypes circulating in populations have importance both for epidemiological data and selection of vaccine strains.


title: Development and comparison of the real-time amplification based methods--NASBA-Beacon, RT-PCR taqman and RT-PCR hybridization probe assays--for the qualitative detection of sars coronavirus.

text: The aim of this study was to develop a rapid, sensitive and robust procedure for the qualitative detection of SARS coronavirus RNA. Three unique detection formats were developed for real-time RNA amplification assays: a post amplification detection step with a virus-specific internal capture probe based on Taqman (RT-PCR TaqMan assay), hybridization probe (RT-PCR hybridization probe assay) and a real-time assay with virus-specific molecular beacon probes (NASBA-Beacon assay). The analytical sensitivity or reproducibility of the test results among those three assays was compared. All assays yielded results by detecting SARS coronavirus targeting the BNI-1 region in less than 2 hours. RNA detection by all the formats was unaffected by the presence of human sputum. The limits of detection were at least 10 copies of input RNA for both RT-PCR formats (RT-PCR TaqMan and RT-PCR hybridization probe assays), while the NASBA-Beacon assay could detect as little as 1 copy per reaction, with high reproducibility of the coefficient of variation (CV) of <10. These results demonstrate that real-time NASBA provides a rapid and sensitive alternative to RT-PCR for the routine qualitative assay of sputum for SARS corona viral RNA detection.


Differential diagnostic blocking ELISA for use in conjunction with modified-live IBRV gIII-deleted marker vaccine.


title: 2-D graphical representation of protein sequences and its application to coronavirus phylogeny.

text: Based on a five-letter model of the 20 amino acids, we propose a new 2-D graphical representation of protein sequence. Then we transform the 2-D graphical representation into a numerical characterization that will facilitate quantitative comparisons of protein sequences. As an application, we construct the phylogenetic tree of 56 coronavirus spike proteins. The resulting tree agrees well with the established taxonomic groups.


title: Nuclear localization of non-structural protein 1 and nucleocapsid protein of equine arteritis virus.

text: RNA synthesis (genome replication and subgenomic mRNA transcription) directed by equine arteritis virus (EAV; family Arteriviridae, order Nidovirales) occurs on modified cytoplasmic membranes to which most viral replicase subunits localize. Remarkably, a fraction of non-structural protein 1 (nsp1), a protein essential for transcription but dispensable for genome replication, is present in the host cell nucleus, in particular during the earlier stages of infection. Expression of GFP-tagged fusion proteins revealed that nsp1 is actively imported into the nucleus. Although the signals responsible for nsp1 transport could not be identified, our studies revealed that another EAV protein with a partially nuclear localization, the nucleocapsid (N) protein, utilizes the CRM1-mediated nuclear export pathway. Inactivation of this pathway with the drug leptomycin B resulted in the unexpected and immediate nuclear retention of all N protein molecules, thus revealing that the protein shuttles between cytoplasm and nucleus before playing its role in cytoplasmic virus assembly.


Rotavirus diarrhoea in patient with antibody to human immunodeficiency virus (HIV)


title: Sequence motifs involved in the regulation of discontinuous coronavirus subgenomic RNA synthesis.

text: Coronavirus transcription leads to the synthesis of a nested set of mRNAs with a leader sequence derived from the 5' end of the genome. The mRNAs are produced by a discontinuous transcription in which the leader is linked to the mRNA coding sequences. This process is regulated by transcription-regulating sequences (TRSs) preceding each mRNA, including a highly conserved core sequence (CS) with high identity to sequences present in the virus genome and at the 3' end of the leader (TRS-L). The role of TRSs was analyzed by reverse genetics using a full-length infectious coronavirus cDNA and site-directed mutagenesis of the CS. The canonical CS-B was nonessential for the generation of subgenomic mRNAs (sgmRNAs), but its presence led to transcription levels at least 10(3)-fold higher than those in its absence. The data obtained are compatible with a transcription mechanism including three steps: (i) formation of 5'-3' complexes in the genomic RNA, (ii) base-pairing scanning of the nascent negative RNA strand by the TRS-L, and (iii) template switching during synthesis of the negative strand to complete the negative sgRNA. This template switch takes place after copying the CS sequence and was predicted in silico based on high base-pairing score between the nascent negative RNA strand and the TRS-L and minimum DeltaG.


title: Development of a nested PCR assay for detection of feline infectious peritonitis virus in clinical specimens.

text: A diagnostic test for feline infectious peritonitis virus (FIPV) infection based on a nested PCR (nPCR) assay was developed and tested with FIPV, feline enteric coronavirus (FECV), canine coronavirus (CCV), and transmissible gastroenteritis virus (TGEV) and clinical fluid samples from cats with effusive feline infectious peritonitis (FIP). The target sequence for the assay is in the S1 region of the peplomer protein E2 gene. A vaccine strain of FIPV and two wild-type FIPV strains tested positive, but FECV, TGEV, and CCV tested negative. Preliminary tests with 12 cats with clinical evidence of effusive FIP and 11 cats with an illness associated with effusions, but attributed to other causes, were performed. Eleven of the 12 cats with effusive FIP tested positive, while 1 was negative. Ten of the 11 cats ill from other causes tested negative, while 1 was positive. On the basis of clinical laboratory and histopathologic criteria, the preliminary sensitivity and specificity of the assay were 91.6 and 94%, respectively.