title: Oral exposure to diabetes-promoting food or immunomodulators in neonates alters gut cytokines and diabetes.

text: Disease development in diabetes-prone BB rats is modified by the type of diet fed after weaning. The aim of this investigation was to determine whether exposure during the first week of life to antigens from a known diabetes-promoting diet (NIH-07) could modify diabetes incidence and, if so, to what extent this occurs via alterations in systemic T-cell reactivity, gut cytokines, or islet infiltration. Diabetes-prone BB (BBdp) rats were hand-fed twice daily between age 4 and 7 days with vehicle, a hydrolyzed casein (HC)-based infant formula, Pregestimil (PG), PG + cereal-based NIH-07 diet, PG + lipopolysaccharides (LPS) or PG + LPS + silica. After weaning, they were fed either an NIH-07 diet or a semipurified HC (diabetes-retardant) diet until 150 days. In separate studies, 5-day-old BBdp rat pups were administered the aforementioned treatments, and expression of intestinal mRNA for gamma-interferon (IFN-gamma) or transforming growth factor-beta (TGF-beta) was quantified using reverse transcriptase-polymerase chain reaction. The effect of early oral treatment with NIH-07 or PG on systemic T-cell reactivity was evaluated using footpad swelling delayed-type hypersensitivity (DTH) and the popliteal lymph node assay. Oral exposure of neonates to a complex mixture of antigens from the diabetes-promoting diet delayed onset of diabetes (79 vs. 88 days) and prevented disease in approximately one-third of animals. A similar protective effect was seen for neonatal exposure to wheat gluten in animals subsequently weaned onto a semipurified wheat gluten diet. By contrast, LPS-treated neonates displayed more severe insulitis and developed diabetes at an increased rate, which was significantly suppressed by co-administration of silica particles. The protective effect of early exposure to diabetogenic diets was not associated with significant reduction of islet infiltration, and there was no impact on the DTH response to food antigens. However, whereas diabetes-resistant BBc rats developed systemic tolerance to NIH-07 antigens fed chronically, BBdp rats did not. The lack of effect of the early oral antigen regimen on the DTH reaction in the footpad, a classic Th1-mediated reaction, suggests little effect on systemic T-cell reactivity. However, local effects were observed in the small intestine. Oral exposure to diabetes-promoting food antigens or LPS downregulated the Th1 cytokine IFN-gamma and decreased the IFN-gamma/TGF-beta ratio. Thus, oral exposure to diabetes-promoting food antigens and immune modulators in neonates can modify diabetes expression in association with changes in local cytokine balance in the gut.


Middle East respiratory syndrome coronavirus and the upcoming Hajj.


SNMMI COVID-19 Resource Center.


title: A comparison of infection control program resources, activities, and antibiotic resistant organism rates in Canadian acute care hospitals in 1999 and 2005: pre- and post-severe acute respiratory syndrome.

text: INTRODUCTION The Resources for Infection Control in Hospitals (RICH) project assessed infection control programs and rates of antibiotic-resistant organisms (AROs) in Canadian acute care hospitals in 1999. In the meantime, the Severe Acute Respiratory Syndrome (SARS) outbreak and the concern over pandemic influenza have stimulated considerable government and healthcare institutional efforts to improve infection control systems in Canada. METHODS In 2006, a version of the RICH survey similar to the original RICH instrument was mailed to infection control programs in all Canadian acute care hospitals with 80 or more beds. Chi-square, ANOVA, and analysis of covariance analyses tested for differences between the 1999 and 2005 samples for infection control program components and ARO rates. RESULTS 72.3% of Canadian acute care hospitals completed the RICH survey for 1999 and 60.1% for 2005. Hospital size was controlled for in analyses involving AROs and surveillance and control intensity levels. Methicillin-resistant Staphylococcus aureus (MRSA) rates increased from 1999 to 2005 (F = 9.4, P = 0.003). In 2005, the MRSA rate was 5.2 (SD 6.1) per 1,000 admissions and in 1999 was 2.0 (SD 2.9). Clostridium difficile-associated diarrhea (CDAD) rates, trended up from 1999 to 2005 (F = 2.9, P = 0.09). In 2005, the mean CDAD rate was 4.7 (SD 4.3) and in 1999 it was 3.8 (SD 4.3). The proportion of hospitals that reported having new nosocomial Vancomycin-resistant Enterococcus (VRE) cases was greater in 2005 than in 1999 (X = 10.5, P = 0.001). In 1999, 34.5% (40 of 116) hospitals reported having new nosocomial VRE cases and in 2005, 61.0% (64 of 105) reported new cases. Surveillance intensity index scores increased from 61.7 (SD 18.5) in 1999 to 68.1 (SD 15.4) in 2005 (F = 4.1, P = 0.04). Control intensity index scores, trended upwards slightly from 60.8 (SD 14.6) in 1999 to 64.1 (12.2) in 2005 (F = 3.2, P = 0.07). ICP full time equivalents (FTEs) per 100 beds increased from 0.5 (SD 0.2) in 1999 to 0.8 (SD 0.3) in 2005 (F = 90.8, P < 0.0001). However, the proportion of ICPs in hospitals certified by the Certification Board of Infection Control (CBIC) decreased from 53% (SD 46) in 1999 to 38% (SD 36) in 2005 (F = 8.7, P = 0.004). CONCLUSIONS Canadian infection control programs in 2005 continued to fall short of expert recommendations for human resources and surveillance and control activities. Meanwhile, Nosocomial MRSA rates more than doubled between 1999 and 2005 and hospitals reporting new nosocomial VRE cases increased 77% over the same period. While investments have been made towards infection control programs in Canadian acute hospitals, the rapid rise in ICP positions has not yet translated into marked improvements in surveillance and control activities. In the face of substantial increases in ARO rates in Canada, continued efforts to train ICPs and support hospital infection control programs are necessary.


title: Achievements and challenges in antiviral drug discovery.

text: The last 40 years have seen the development of several antiviral drugs with therapeutic value in treating life-threatening or debilitating diseases such as those caused by HIV, hepatitis B virus, herpesviruses (such as herpes simplex virus and varicella zoster virus) and influenza virus. These relatively recent advances have been due to technical breakthroughs in the cultivation of viruses in the laboratory, identification of viral enzymes and, more recently, their molecular biology. We describe here the antecedence of several of the existing antivirals and their strengths and weaknesses. We indicate where the major challenges lie for future improvements of current therapies and possible new indications, such as hepatitis C virus and papillomavirus. We also describe how current antiviral therapies are restricted to a rather limited number of viral diseases of sufficient interest to the pharmaceutical industry. Finally we describe the potential threat of emerging viruses and bio-weapons and the challenges that they present to therapy.


title: 18F-FDG PET in sarcoidosis: an observational study in 12 patients treated with infliximab.

text: BACKGROUND 18F-FDG PET is a promising technique in sarcoidosis imaging, although it is not incorporated in routine activity assessment. The purpose of this study was to correlate 18F-FDG PET with standard sarcoidosis activity parameters during infliximab treatment. METHODS Twelve patients with refractory sarcoidosis were treated with 6 cycles of infliximab. Pre- and post-therapy 18F-FDG PET was visually evaluated and SUVmax was measured. In addition, the effect of infliximab was evaluated by changes in symptoms, angiotensin converting enzyme (ACE), soluble interleukin-2 receptor (sIL-2R), vital capacity (VC), diffusion capacity of the lung for carbon monoxide (DLCO) and chest radiography. SUVmax and conventional parameters were correlated. RESULTS Clinical improvement as judged by conventional parameters was seen in all patients, though with a minor response in one. Symptoms improved in 11/12 patients while chest radiographic stages did not change. The decrease in ACE was 39% and in sIL-2R 47% (p<0.01). Improvement of VC and DLCO was 5.4% and 3.3% (p<0.05), respectively. 18F-FDG PET revealed either improvement or normalization in 11/12 (92%) clinically responding patients. The overall decrease in SUVmax was 55% (p<0.01); the patient with a limited response showed a 34% increase. A decrease in SUVmax of the lung parenchyma correlated with an improvement of VC (r=-0.75, p<0.01). No significant correlation between SUVmax and other parameters was found. CONCLUSION Changes imaged by 18F-FDG PET during infliximab treatment in sarcoidosis patients correlate with signs of clinical improvement to a considerate extent, which supports the hypothesis that 18F-FDG uptake represents disease activity.


title: Capnocytophaga sp. isolated from a cat with chronic sinusitis and rhinitis.

text: A Capnocytophaga sp. was inadvertently isolated from a cat with chronic sinusitis and rhinitis when cytopathic effects were observed in Crandall-Reese feline kidney cells that had been inoculated with oropharyngeal swab samples. Although Capnocytophaga spp. are of considerable zoonotic importance, their clinical relevance for dogs or cats has not been established. However, failure to do so may be attributed to the infrequent use of specialized isolation techniques that are required to grow Capnocytophaga spp. To our knowledge, successful isolation of these organisms from a cat with nasopharyngeal disease has not been reported.


title: A pilot study using preoperative cerebral tissue oxygen saturation to stratify cardiovascular risk in major non-cardiac surgery.

text: This prospective pilot study evaluated whether low preoperative cerebral tissue oxygen saturation is associated with unfavourable outcomes after major elective non-cardiac surgery. Eighty-one patients over 60 years of age, American Society of Anesthesiologists physical status 3 or 4, were recruited. Resting cerebral tissue oxygen saturation was recorded on room air, and after oxygen supplementation, using cerebral oximetry. The primary outcome was 30-day major adverse event of combined mortality or severe morbidity, and the secondary outcome was 30-day new disability. Eleven patients (13.6%) suffered a major adverse event, and 28 patients (34.6%) experienced new disability. Room air cerebral tissue oxygen saturation was significantly different between patients who had a major adverse event, 67% (95% confidence interval [CI] 65-70) versus unaffected, 71% (95% CI 70-72; P=0.04). No statistical difference was found between patients for new disability (range 70%-74%; P=0.73). Room air cerebral tissue oxygen saturation was significantly associated with major adverse events (odds ratio 1.36 (95% CI 1.03-1.79), P=0.03). Saturation levels ≤68% carried a positive likelihood ratio of 2.2 for death or severe morbidity, P=0.04. A definitive trial is required to confirm if cerebral oximetry can be used to stratify the cardiovascular risk of patients presenting for non-cardiac surgery.


title: Long-term impact on a closed household of pet cats of natural infection with feline coronavirus, feline leukaemia virus and feline immunodeficiency virus.

text: A closed household of 26 cats in which feline coronavirus (FCoV), feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) were endemic was observed for 10 years. Each cat was seropositive for FCoV on at least one occasion and the infection was maintained by reinfection. After 10 years, three of six surviving cats were still seropositive. Only one cat, which was also infected with FIV, developed feline infectious peritonitis (FIP). Rising anti-FCoV antibody titres did not indicate that the cat would develop FIP. The FeLV infection was self-limiting because all seven of the initially viraemic cats died within five years and the remainder were immune. However, FeLV had the greatest impact on mortality. Nine cats were initially FIV-positive and six more cats became infected during the course of the study, without evidence of having been bitten. The FIV infection did not adversely affect the cats' life expectancy.


title: Antiviral antibodies are necessary to prevent cytotoxic T-lymphocyte escape in mice infected with a coronavirus.

text: Mutation within virus-derived CD8 T-cell epitopes can effectively abrogate cytotoxic T-lymphocyte (CTL) recognition and impede virus clearance in infected hosts. These so-called "CTL escape variant viruses" are commonly selected during persistent infections and are associated with rapid disease progression and increased disease severity. Herein, we tested whether antiviral antibody-mediated suppression of virus replication and subsequent virus clearance were necessary for preventing CTL escape in coronavirus-infected mice. We found that compared to wild-type mice, B-cell-deficient mice did not efficiently clear infectious virus, uniformly developed clinical disease, and harbored CTL escape variant viruses. These data directly demonstrate a critical role for antiviral antibody in protecting from the selective outgrowth of CTL escape variant viruses.


title: Type IVB pilus operon promoter controlling expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid gene in Salmonella enterica Serovar Typhi elicits full immune response by intranasal vaccination.

text: Attenuated Salmonella enterica serovar Typhi strains have been considered to be attractive as potential live oral delivery vector vaccines because of their ability to elicit the full array of immune responses in humans. In this study, we constructed an attenuated S. enterica serovar Typhi strain stably expressing conserved nucleocapsid (N) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) by integrating the N gene into the pilV gene, which was under the control of the type IVB pilus operon promoter in S. enterica serovar Typhi. BALB/c mice were immunized with this recombinant strain through different routes: intranasally, orogastrically, intraperitoneally, and intravenously. Results showed that the intranasal route caused the highest production of specific immunoglobulin G (IgG), IgG2a, and secretory IgA, where IgG2a was imprinted as a Th1 cell bias. Moreover, this recombinant live vaccine induced significantly high levels of specific cytotoxic T-lymphocyte activities and increased gamma interferon-producing T cells compared with the parental strain. Our work provides insights into how the type IVB pilus operon promoter controlling SARS-CoV N gene expression in Salmonella might be attractive for a live-vector vaccine against SRAS-CoV infection, for it could induce mucosal, humoral, and cellular immune responses. Our work also indicates that the type IVB pilus operon promoter controlling foreign gene expression in Salmonella can elicit full immune responses by intranasal vaccination.


title: Characterization of the coronavirus M protein and nucleocapsid interaction in infected cells.

text: Coronavirus contains three envelope proteins, M, E and S, and a nucleocapsid, which consists of genomic RNA and N protein, within the viral envelope. We studied the macromolecular interactions involved in coronavirus assembly in cells infected with a murine coronavirus, mouse hepatitis virus (MHV). Coimmunoprecipitation analyses demonstrated an interaction between N protein and M protein in infected cells. Pulse-labeling experiments showed that newly synthesized, unglycosylated M protein interacted with N protein in a pre-Golgi compartment, which is part of the MHV budding site. Coimmunoprecipitation analyses further revealed that M protein interacted with only genomic-length MHV mRNA, mRNA 1, while N protein interacted with all MHV mRNAs. These data indicated that M protein interacted with the nucleocapsid, consisting of N protein and mRNA 1, in infected cells. The M protein-nucleocapsid interaction occurred in the absence of S and E proteins. Intracellular M protein-N protein interaction was maintained after removal of viral RNAs by RNase treatment. However, the M protein-N protein interaction did not occur in cells coexpressing M protein and N protein alone. These data indicated that while the M protein-N protein interaction, which is independent of viral RNA, occurred in the M protein-nucleocapsid complex, some MHV function(s) was necessary for the initiation of M protein-nucleocapsid interaction. The M protein-nucleocapsid interaction, which occurred near or at the MHV budding site, most probably represented the process of specific packaging of the MHV genome into MHV particles.


The risk of a pandemic with the influenza A (H1N1) virus.


title: [The biological characteristics of SARS virus and its related coronaviruses].

text: Cases of the life-threatening respiratory disease with no identified cause (designated as "severe acute respiratory syndrome", SARS, in March 2003) were first reported in late 2002 from Guangdong Province, China; they were followed by reports from about other 30 countries (or regions) such as Vietnam, Singapore, Thailand, Hong Kong (China), Canada, and USA etc. Because of its ongoing epidemic and high death rate, SARS has shined an intense spotlight all over the world. The World Health Organization (WHO) has promptly established a network of international laboratories consisting of 13 members around the 10 countries to facilitate the identification of the causative agent of SARS. A novel coronavirus, SARS virus, fulfilling all of Koch's postulates was announced to be the primary aetiological agent of SARS on April 16 by WHO shortly after the Canadian scientists released the full-length genome sequence of SARS virus (Tor2) on April 12. China is now facing a formidable task to fight SARS. In this article, we present a brief summary on the biological characteristics of coronavirus with its associated diseases, and make some suggestions on how to curb this outbreak and how to cure SARS disease based on the potential targets of this novel virus.


title: Rapid and efficient preparation of monoclonal antibodies against SARS-associated coronavirus nucleocapsid protein by immunizing mice.

text: OBJECTIVE To develop a rapid and efficient method for preparing monoclonal antibodies (mAb) against SARS-associated coronavirus (SARS-Cov) nucleocapsid (N) protein. METHODS BALB/c mice were injected with the recombinant N protein of SARS-Cov into the foot-pads for the immunization, and the popliteal lymph nodes were isolated 15 d later for mAb-producing hybridomas, from which the mAbs against the N protein of SARS-Cov were screened. The identification of the mAb against the N protein of SARS-Cov was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. RESULTS Four strains of hybridomas were obtained that produced the mAb specific to the N protein without detectable cross-reactivity with other pathogens. Of the 4 strains, 2 were identified as the immunoglobulin G1 (IgG1) isotype, 1 IgG2a, and the other IgG2b, with affinity constants (Ka) of 2 of the strains being 4.14 x 10(-9)M and 3.19 x 10(-9)M respectively. CONCLUSION This is the first report on the preparation of mAb that is specific to the SARS-Cov, and the high-specificity and high-affinity mAb produced by the 4 strains of hybridomas provide a basis for further researches on the pathogenesis and early diagnosis of SARS.


title: The severe acute respiratory syndrome coronavirus nucleocapsid protein is phosphorylated and localizes in the cytoplasm by 14-3-3-mediated translocation.

text: The severe acute respiratory syndrome coronavirus(SARS-CoV) nucleocapsid (N) protein is one of the four structural proteins of the virus and is predicted to be a 46-kDa phosphoprotein. Our in silico analysis predicted N to be heavily phosphorylated at multiple residues. Experimentally, we have shown in this report that the N protein of the SARS-CoV gets serine-phosphorylated by multiple kinases, in both the cytoplasm and the nucleus. The phosphoprotein is stable and localizes in the cytoplasm and coprecipitates with the membrane fraction. Also, using specific inhibitors of phosphorylation and an in vitro phosphorylation assay, we show that the nucleocapsid protein is a substrate of cyclin-dependent kinase (CDK), glycogen synthase kinase, mitogen-activated protein kinase, and casein kinase II. Further, we show that the phosphorylated protein is translocated to the cytoplasm by binding to 14-3-3 (tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein). 14-3-3 proteins are a family of highly conserved, ubiquitously expressed eukaryotic proteins that function primarily as adapters that modulate interactions between components of various cellular signaling and cell cycle regulatory pathways through phosphorylation-dependent protein-protein interactions. Coincidentally, the N protein was also found to downregulate the expression of the theta isoform of 14-3-3 (14-3-3theta), leading to the accumulation of phosphorylated N protein in the nucleus, in the absence of growth factors. Using short interfering RNA specific to 14-3-3theta we have inhibited its expression to show accumulation of phosphorylated N protein in the nucleus. Thus, the data presented here provide a possible mechanism for phosphorylation-dependent nucleocytoplasmic shuttling of the N protein. This 14-3-3-mediated transport of the phosphorylated N protein and its possible implications in interfering with the cellular machinery are discussed.


title: Role for nsP2 proteins in the cessation of alphavirus minus-strand synthesis by host cells.

text: In order to establish nonlytic persistent infections (PI) of BHK cells, replicons derived from Sindbis (SIN) and Semliki Forest (SFV) viruses have mutations in nsP2. Five different nsP2 PI replicons were compared to wild-type (wt) SIN, SFV, and wt nsPs SIN replicons. Replicon PI BHK21 cells had viral RNA synthesis rates that were less than 5% of those of the wt virus and approximately 10% or less of those of SIN wt replicon-infected cells, and, in contrast to wt virus and replicons containing wt nsP2, all showed a phenotype of continuous minus-strand synthesis and of unstable, mature replication/transcription complexes (RC+) that are active in plus-strand synthesis. Minus-strand synthesis and incorporation of [3H]uridine into replicative intermediates differed among PI replicons, depending on the location of the mutation in nsP2. Minus-strand synthesis by PI cells appeared normal; it was dependent on continuous P123 and P1234 polyprotein synthesis and ceased when protein synthesis was inhibited. The failure by the PI replicons to shut off minus-strand synthesis was not due to some defect in the PI cells but rather was due to the loss of some function in the mutated nsP2. This was demonstrated by showing that superinfection of PI cells with wt SFV triggered the shutdown of minus-strand synthesis, which we believe is a host response to infection with alphaviruses. Together, the results indicate alphavirus nsP2 functions to engage the host response to infection and activate a switch from the early-to-late phase. The loss of this function leads to continuous viral minus-strand synthesis and the production of unstable RC+.


title: Functional role of hepatitis C virus chimeric glycoproteins in the infectivity of pseudotyped virus.

text: The putative envelope glycoproteins of hepatitis C virus (HCV) likely play an important role in the initiation of viral infection. Available information suggests that the genomic regions encoding the putative envelope glycoproteins, when expressed as recombinant proteins in mammalian cells, largely accumulate in the endoplasmic reticulum. In this study, genomic regions which include the putative ectodomain of the E1 (amino acids 174 to 359) and E2 (amino acids 371 to 742) glycoproteins were appended to the transmembrane domain and cytoplasmic tail of vesicular stomatitis virus (VSV) G protein. This provided a membrane anchor signal and the VSV incorporation signal at the carboxy termini of the E1 and E2 glycoproteins. The chimeric gene constructs exhibited expression of the recombinant proteins on the cell surface in a transient expression assay. When infected with a temperature-sensitive VSV mutant (ts045) and grown at the nonpermissive temperature (40.5 degrees C), cells transiently expressing the E1 or E2 chimeric glycoprotein generated VSV/HCV pseudotyped virus. The resulting pseudotyped virus generated from E1 or E2 surprisingly exhibited the ability to infect mammalian cells and sera derived from chimpanzees immunized with the homologous HCV envelope glycoproteins neutralized pseudotyped virus infectivity. Results from this study suggested a potential functional role for both the E1 and E2 glycoproteins in the infectivity of VSV/HCV pseudotyped virus in mammalian cells. These observations further suggest the importance of using both viral glycoproteins in a candidate subunit vaccine and the potential for using a VSV/HCV pseudotyped virus to determine HCV neutralizing antibodies.


title: Severe acute respiratory syndrome diagnostics using a coronavirus protein microarray.

text: To monitor severe acute respiratory syndrome (SARS) infection, a coronavirus protein microarray that harbors proteins from SARS coronavirus (SARS-CoV) and five additional coronaviruses was constructed. These microarrays were used to screen approximately 400 Canadian sera from the SARS outbreak, including samples from confirmed SARS-CoV cases, respiratory illness patients, and healthcare professionals. A computer algorithm that uses multiple classifiers to predict samples from SARS patients was developed and used to predict 206 sera from Chinese fever patients. The test assigned patients into two distinct groups: those with antibodies to SARS-CoV and those without. The microarray also identified patients with sera reactive against other coronavirus proteins. Our results correlated well with an indirect immunofluorescence test and demonstrated that viral infection can be monitored for many months after infection. We show that protein microarrays can serve as a rapid, sensitive, and simple tool for large-scale identification of viral-specific antibodies in sera.


As the bat flies.


title: Current vaccine adjuvants: an overview of a diverse class.

text: Numerous compounds are under evaluation as immunological adjuvants for improvement of vaccine performance. This review will briefly summarize some of the many diverse substances that are currently being utilized as vaccine adjuvants in preclinical and clinical studies.


title: Soft-tissue Defects After Total Knee Arthroplasty: Management and Reconstruction.

text: Wound healing complications associated with total knee arthroplasty present a considerable challenge to the orthopaedic surgeon. To ensure preservation of a functional joint, the management of periprosthetic soft-tissue defects around the knee requires rapid assessment, early and aggressive débridement, and durable, contoured coverage. Several reconstructive options are available to tailor soft-tissue coverage to the location, size, and depth of the wound. Special consideration should be given to the timing of the intervention, management of infection, and prosthesis salvage. The merits of each reconstructive option, including perforator, fasciocutaneous, muscular, and free microvascular flaps, should be weighed to select the most appropriate option. The proposed approach can guide surgeons in treating patients with these complex soft-tissue defects.


title: Talk to Patients About: Why Will It Take So Long for a COVID-19 Vaccine?

text: Of all the solutions to the COVID-19 pandemic, a vaccine is the most attractive. With just one shot, people could transform the disease from a global disruptor to an easily manageable public health problem. But vaccines can't be whipped up overnight.


title: Thrombus in Transit and Impending Pulmonary Embolism Detected on POCUS in a Patient with COVID-19 Pneumonia.

text: Coronavirus disease 2019 (COVID-19) is a pandemic that started in China in December 2019 and carries a high risk of morbidity and mortality. To-date (4-22-2020) it affected over 2.6 million people and resulted in nearly 200,000 death worldwide mainly due to severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2). Among the major underlying pathophysiologic mechanisms in COVID 19 is hypercoagulability, leading to increased risk for deep vein thrombosis and pulmonary embolism that contribute to increased morbidity and mortality. In this report, we present the case of a 55-year-old man who presented with COVID-19 pneumonia, and was found to have a thrombus in transit by routine point of care ultrasound (POCUS). While computer tomography (CT) angiography is the test of choice, the utilization of point of care ultrasound (POCUS) has gained traction as an adjunctive means of surveillance for the development of VTE in patients with COVID-19. In this report, we discuss the clinical utility of POCUS in diagnosing thrombus in transit in COVID 19 populations.


title: Comparison of effectiveness of whole viral, N and N199 proteins by ELISA for the rapid diagnosis of severe acute respiratory syndrome coronavirus.

text: BACKGROUND Although severe acute respiratory syndrome (SARS) has been controlled, the subsequently emerging sporadic cases in 2004 emphasize the necessity of developing a rapid diagnostic method, which would be of great help in clinical diagnosis and also wild host screening. This study aims to establish an effective and rapid serological tool for the diagnosis of SARS-CoV by comparison among whole viral, N and N199 proteins by ELISA. METHODS SARS-CoV N and N199 (a truncated nucleocapsid gene) genes were cloned, expressed, identified by Western blotting, and applied in screening of human and swine samples. Sera of SARS convalescent-phase patients, normal human sera, sera of patients with other respiratory diseases, and swine sera were screened by ELISA, with whole SARS-CoV F69, N and N199 proteins as antigens. RESULTS The sensitivity and specificity of N and N199 proteins in human sera diagnosis were approximate (P = 0.743), which was higher than whole viral protein but the difference was not significant (P = 0.234). The N199 protein proved to be more specific in swine sera screening than whole viral and N protein (P < 0.001). CONCLUSION N199 protein is feasible in both clinical diagnosis and SARS-CoV reservoir screening.


title: SARS commission reports on the Canadian outbreak.

text: The third and final report investigating the 2003 Toronto outbreak of severe acute respiratory syndrome (SARS) that led to 44 deaths says hospital downsizing was a factor and warns that 'serious problems persist and much needs to be done'.


title: Immunohistochemical identification of porcine respiratory coronavirus antigen in the lung of conventional pigs.

text: Nine 3-week-old conventional pigs were inoculated intranasally with a Korean isolate of porcine respiratory coronavirus (PRCV). Three 3-week-old conventional pigs were kept as noninoculated controls. Three inoculated and one control pig were euthanatized at 5, 10, and 15 days postinoculation (DPI), respectively. All nine inoculated pigs developed respiratory disease. Interstitial pneumonia characterized by mononuclear inflammatory cell infiltration into alveolar septa were seen microscopically in all nine PRCV-infected pigs. Histopathologic changes were severe at DPI, modest at 10 DPI, and almost resolved at 15 DPI. PRCV antigen was not detected in bronchial and bronchiolar epithelium but was detected in interstitial macrophages in the lungs. This Korean isolate of PRCV appeared to replicate primarily within alveolar macrophages in the respiratory system of the pig.


title: O-glycosylation of the coronavirus M protein. Differential localization of sialyltransferases in N- and O-linked glycosylation.

text: It has previously been shown that the M (E1) glycoprotein of mouse hepatitis virus strain A59 (MHV-A59) contains only O-linked oligosaccharides and localizes to the Golgi region when expressed independently. A detailed pulse-chase analysis was made of the addition of O-linked sugars to the M protein; upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, three different electrophoretic forms could be distinguished that corresponded to the sequential acquisition of N-acetylgalactosamine (GalNAc), galactose (Gal), and sialic acid (SA). A fourth and fifth form could also be detected which we were unable to identify. Following Brefeldin A treatment, the M protein still acquired GalNAc, Gal, and SA, but the fourth and fifth forms were absent, suggesting that these modifications occur in the trans-Golgi network (TGN). In contrast, in the presence of BFA, the G protein of vesicular stomatitis virus (VSV), which contains N-linked oligosaccharides, acquired Gal and fucose but not SA. These results are consistent with earlier published data showing that Golgi compartments proximal to the TGN, but not the TGN itself, relocate to the endoplasmatic reticulum/intermediate compartment. More importantly, our data argue that, whereas addition of SA to N-linked sugars occurs in the TGN the acquisition of both SA on O-linked sugars and the addition of fucose to N-linked oligosaccharides must occur in Golgi compartments proximal to the TGN. The glycosylation of the M protein moreover indicates that it is transported to trans-Golgi and TGN. This was confirmed by electron microscopy immunocytochemistry, showing that the protein is targeted to cisternae on the trans side of the Golgi and co-localizes, at least in part, with TGN 38, a marker of the TGN, as well as with a lectin specific for sialic acid.


title: Aetiology of acute gastroenteritis in infancy and early childhood in southern India.

text: The aetiology of acute gastroenteritis was studied in 50 infants and young children. Bacterial pathogens were isolated in 33, enteropathogenic E. coli (EPEC), Salmonella, and Shigella being the commonest isolates. Rotaviruses were detected in the stools of 13 of the cases. All children with gastroenteritis in whom rotavirus was detected were seen during the months July to December. In 30 children who served as controls, EPEC were isolated in 6, but rotavirus was detected in none. It is concluded that infection with rotaviruses is a significant cause of morbidity in children with gastroenteritis in southern India.


title: Neonatal calf diarrhea caused by a virus that induces villous epithelial cell syncytia.

text: Intestinal lesions caused by a virus serologically unrelated to the calf diarrheal rotavirus or coronavirus were studied in gnotobiotic calves. The virion purified from feces from infected calves was a fringed particle with a diameter of about 100 nm. The incubation period from time of inoculation per orum to onset of diarrhea in calves was as short as 8 hours. The viral infection in bacteria-free calves or calves not contaminated with pathogenic bacteria caused severe illness for only 24 hours. When bacteria such as the K99 antigen Escherichia coli were present, the combined infection caused mortality. Lesions occurred only in the small intestinal villous epithelium. Calves euthanatized shortly before or after the onset of diarrhea had developed villous epithelial cell syncytia that contained numberous virions in the cytoplasm. Within 2 to 3 hours after onset of diarrhea, the infected cells were shed and the villi had denuded tips or had cuboidal to squamous epithelial cells.


title: Subunit composition and conformational stability of the oligomeric form of the avian reovirus cell-attachment protein sigmaC.

text: Previous work has shown that the avian reovirus cell-attachment sigma C (sigmaC) protein is a multimer. In the first part of this study the oligomerization state of intracellularly synthesized sigmaC was analysed by different approaches, including SDS-PAGE, chemical cross-linking, sedimentation and gel filtration analysis. All these approaches indicated that protein sigmaC in its native state is a homotrimer. In the second part of the present work we investigated the effect of different factors and reagents on oligomer stability, in order to elucidate the nature of the forces that maintain the conformational stability of the homotrimer. Our results, based on the stabilizing effect conferred by reducing agents, demonstrate that the sigmaC subunits are not covalently bound via disulfide linkages. They further suggest that the formation of an intrachain disulfide bond between the two cysteine residues of the sigmaC polypeptide has a negative effect on oligomer stability. The susceptibility of the trimer to pH, temperature, ionic strength, chemical denaturants and detergents indicates that hydrophobic interactions contribute much more to oligomer stability than do ionic interactions and hydrogen bonding. Finally, our results also reveal that mammalian and avian reovirus cell attachment proteins follow different subunit dissociation pathways.


title: A 9,000-year record of Chagas' disease.

text: Tissue specimens from 283 principally spontaneously (naturally) desiccated human mummies from coastal and low valley sites in northern Chile and southern Peru were tested with a DNA probe directed at a kinetoplast DNA segment of Trypanosoma cruzi. The time interval spanned by the eleven major cultural groups represented in the sample ranged from approximately 9,000 years B.P. (7050 B.C.) to approximately the time of the Spanish conquest, approximately 450 B.P. ( approximately 1500 A.D.). Forty-one percent of the tissue extracts, amplified by the PCR reacted positively (i.e., hybridized) with the probe. Prevalence patterns demonstrated no statistically significant differences among the individual cultural groups, nor among subgroups compared on the basis of age, sex, or weight of specimen tested. These results suggest that the sylvatic (animal-infected) cycle of Chagas' disease was probably well established at the time that the earliest humans (members of the Chinchorro culture) first peopled this segment of the Andean coast and inadvertently joined the many other mammal species acting as hosts for this parasite.


The 1918-1919 Influenza Pandemic and National Amnesia.


title: Cellular receptors for transmissible gastroenteritis virus on porcine enterocytes.

text: The activity of aminopeptidase-N (APN), reported to be a major receptor for porcine transmissible gastroenteritis virus (TGEV), in enterocyte fractions harvested from the jejunal villi and crypts of newborn and weaned piglets, did not correspond with the levels of saturable virus binding previously demonstrated for the same fractions. Plasma membranes prepared from enterocytes harvested from the jejunal villi of a newborn piglet were used in the preparation of a monoclonal antibody (MAb) which blocked the binding of TGEV, but not that of the porcine respiratory coronavirus (PRCV), to ST cells. This MAb immunoprecipitated a 200 kDa non-glycosylated protein from lysates of ST cells, which was not precipitated by an anti-APN MAb. The 200 kDa protein was shown by immunostaining and fluorescence activated cell scanning to be present on ST cells and on villous enterocytes from newborn piglets, but not on MDBK cells or enterocytes from weaned piglets. APN was demonstrated by the same techniques to be present on villous enterocytes from both newborn and weaned piglets, as well as on ST cells. It was concluded that the 200 kDa protein may be a second receptor for TGEV, contributing to the high susceptibility of newborn piglets to the virus.


title: Coronavirus E1 glycoprotein expressed from cloned cDNA localizes in the Golgi region.

text: Cloned cDNA encoding the membrane glycoprotein E1 of the coronavirus mouse hepatitis virus strain A59 was expressed transiently in a monkey fibroblast cell line (COS) by using a simian virus 40-based vector. As determined by indirect immunofluorescence microscopy, the E1 protein accumulated intracellularly in a perinuclear region coincident with a Golgi marker. The same three species of E1 that occur in virus-infected cells were also found in transfected cells. These are one unglycosylated form and two apparently O-glycosylated forms that could be labeled in a tunicamycin-resistant fashion with [3H]glucosamine. Because O glycosylation occurs posttranslationally in the Golgi apparatus, we could show, by monitoring the rate of acquisition of oligosaccharides, that the transport of E1 from the rough endoplasmic reticulum to the Golgi apparatus had a half time of between 15 and 30 min.


title: Outbreak of norovirus illness in a college summer camp: impact of cleaning on occurrence of norovirus on fomites.

text: During the summer of 2005 an outbreak of norovirus acute gastroenteritis occurred in a residential college summer camp and was reported to the local health department. The outbreak spread rapidly to several other groups concurrently sharing the same facilities. During the investigation, fomites were sampled at different times in dorm rooms and tested for norovirus. The number of norovirus-positive rooms increased after the first room cleaning, from 40% to 73%. After the initial cleaning, the staff was instructed on proper cleaning and disinfection procedures and provided with disposable disinfecting wipes to reduce cross contamination, and the number of norovirus-positive rooms decreased to 30%. These findings reinforce the need for appropriate cleaning and disinfection procedures during a norovirus outbreak.


title: Small envelope protein E of SARS: cloning, expression, purification, CD determination, and bioinformatics analysis.

text: AIM To obtain the pure sample of SARS small envelope E protein (SARS E protein), study its properties and analyze its possible functions. METHODS The plasmid of SARS E protein was constructed by the polymerase chain reaction (PCR), and the protein was expressed in the E coli strain. The secondary structure feature of the protein was determined by circular dichroism (CD) technique. The possible functions of this protein were annotated by bioinformatics methods, and its possible three-dimensional model was constructed by molecular modeling. RESULTS The pure sample of SARS E protein was obtained. The secondary structure feature derived from CD determination is similar to that from the secondary structure prediction. Bioinformatics analysis indicated that the key residues of SARS E protein were much conserved compared to the E proteins of other coronaviruses. In particular, the primary amino acid sequence of SARS E protein is much more similar to that of murine hepatitis virus (MHV) and other mammal coronaviruses. The transmembrane (TM) segment of the SARS E protein is relatively more conserved in the whole protein than other regions. CONCLUSION The success of expressing the SARS E protein is a good starting point for investigating the structure and functions of this protein and SARS coronavirus itself as well. The SARS E protein may fold in water solution in a similar way as it in membrane-water mixed environment. It is possible that beta-sheet I of the SARS E protein interacts with the membrane surface via hydrogen bonding, this beta-sheet may uncoil to a random structure in water solution.


title: Immunization against feline coronaviruses.

text: Feline infectious peritonitis (FIP) is caused by one of several strains of feline coronaviruses which are grouped into 2 general types of viruses. Infection of cats with FIP virus results in production of serum antibodies which may be protective in conjunction with cell mediated immunity, may provided no protection at all, or may produce an immune enhancement to subsequent exposure to another FIP virus or a recrudescence of the original infecting virus. Attempts at immunization of cats against FIP with inactivated or live FIP viruses have been generally unsuccessful, and often sensitizing the cat through immune enhancement rather than providing protection. Heterologous live virus vaccines using viruses of the same antigenic cluster (transmissible gastroenteritis of swine, canine coronavirus, and human coronavirus 229E) have failed to provide protection against FIP virus. Further research into the exact mechanism of protection and immune enhancement is needed in order to understand ways of producing an effective and safe vaccine.


title: Evaluation of control measures implemented in the severe acute respiratory syndrome outbreak in Beijing, 2003.

text: CONTEXT Beijing, China, experienced the world's largest outbreak of severe acute respiratory syndrome (SARS) beginning in March 2003, with the outbreak resolving rapidly, within 6 weeks of its peak in late April. Little is known about the control measures implemented during this outbreak. OBJECTIVE To describe and evaluate the measures undertaken to control the SARS outbreak. DESIGN, SETTING, AND PARTICIPANTS Data were reviewed from standardized surveillance forms from SARS cases (2521 probable cases) and their close contacts observed in Beijing between March 5, 2003, and May 29, 2003. Procedures implemented by health authorities were investigated through review of official documents and discussions with public health officials. MAIN OUTCOME MEASURES Timeline of major control measures; number of cases and quarantined close contacts and attack rates, with changes in infection control measures, management, and triage of suspected cases; and time lag between illness onset and hospitalization with information dissemination. RESULTS Health care worker training in use of personal protective equipment and management of patients with SARS and establishing fever clinics and designated SARS wards in hospitals predated the steepest decline in cases. During the outbreak, 30 178 persons were quarantined. Among 2195 quarantined close contacts in 5 districts, the attack rate was 6.3% (95% confidence interval [CI], 5.3%-7.3%), with a range of 15.4% (95% CI, 11.5%-19.2%) among spouses to 0.36% (95% CI, 0%-0.77%) among work and school contacts. The attack rate among quarantined household members increased with age from 5.0% (95% CI, 0%-10.5%) in children younger than 10 years to 27.6% (95% CI, 18.2%-37.0%) in adults aged 60 to 69 years. Among almost 14 million people screened for fever at the airport, train stations, and roadside checkpoints, only 12 were found to have probable SARS. The national and municipal governments held 13 press conferences about SARS. The time lag between illness onset and hospitalization decreased from a median of 5 to 6 days on or before April 20, 2003, the day the outbreak was announced to the public, to 2 days after April 20 (P<.001). CONCLUSIONS The rapid resolution of the SARS outbreak was multifactorial, involving improvements in management and triage in hospitals and communities of patients with suspected SARS and the dissemination of information to health care workers and the public.


title: Identification of a domain required for autoproteolytic cleavage of murine coronavirus gene A polyprotein.

text: The 5'-most gene of the murine coronavirus genome, gene A, is presumed to encode viral RNA-dependent RNA polymerase. It has previously been shown that the N-terminal portion of this gene product is cleaved into a protein of 28 kilodaltons (p28). To further understand the mechanism of synthesis of the p28 protein, cDNA clones representing the 5'-most 5.3 kilobases of murine coronavirus mouse hepatitis virus strain JHM were sequenced and subcloned into pT7 vectors from which RNAs were transcribed and translated in vitro. The sequence was found to encode a single long open reading frame continuing from near the 5' terminus of the genome. Although p28 is encoded from the first 1 kilobase at the 5' end of the genome, translation of in vitro-transcribed RNAs indicated that this protein was not detected unless the product of the entire 5.3-kilobase region was synthesized. Translation of RNAs of 3.9 kilobases or smaller yielded proteins which contained the p28 sequence, but p28 was not cleaved. This suggests that the sequence in the region between 3.9 and 5.3 kilobases from the 5' end of the genomic RNA is essential for proteolytic cleavage and contains autoproteolytic activity. The p28 protein could not be cleaved from the smaller primary translation products of gene A, even in the presence of the larger autocleaving protein. Cleavage of the p28 protein was inhibited by addition of the protease inhibitor ZnCl2. This study thus identified a protein domain essential for autoproteolytic cleavage of the gene A polyprotein.


title: Type and quantity of data needed for an early estimate of transmissibility when an infectious disease emerges.

text: An early estimate of disease transmissibility is essential for a well-informed public health response to a newly emerged infectious disease. In this study, we ask what type and quantity of data are needed for useful estimation of the initial reproduction number (R). It is possible to estimate R from case incidence data alone when the growing incidence of cases displays a wave pattern, because the pattern provides information about the serial interval (the time elapsed between the onset of symptoms of a case and symptom onset in individuals infected by that case). When the mode of the serial interval distribution is small, 1.5 days or less, there is generally no informative wave pattern in the observed series of daily incidences. The precision of the estimate of R is then improved substantially by having some observations on the serial interval. For an infectious disease with characteristics such as those of influenza, an estimate of R able to inform plans to mitigate transmission is obtained when the cumulative incidence of cases reaches about 300 and about 10 observations on the serial interval are available.


title: Multistate outbreak of human Salmonella typhimurium infections associated with aquatic frogs - United States, 2009.

text: During April-July 2009, the Utah Department of Health identified five cases of Salmonella Typhimurium infection with indistinguishable pulsed-field gel electrophoresis (PFGE) patterns, predominantly among children. In August, CDC began a multistate outbreak investigation to determine the source of the infections. This report summarizes the results of this ongoing investigation, which, as of December 30, had identified 85 S. Typhimurium human isolates with the outbreak strain from 31 states. In a multistate case-control study, exposure to frogs was found to be significantly associated with illness (63% of cases versus 3% of controls; matched odds ratio [mOR] = 24.4). Among 14 case-patients who knew the type of frog, all had exposure to an exclusively aquatic frog species, the African dwarf frog. Environmental samples from aquariums containing aquatic frogs in four homes of case-patients yielded S. Typhimurium isolates matching the outbreak strain. Preliminary traceback information has indicated these frogs likely came from the same breeder in California. Reptiles (e.g., turtles) and amphibians (e.g., frogs) have long been recognized as Salmonella carriers, and three multistate outbreaks of human Salmonella infections associated with turtle contact have occurred since 2006. However, this is the first reported multistate outbreak of Salmonella infections associated with amphibians. Educational materials aimed at preventing salmonellosis from contact with reptiles should be expanded to include amphibians, such as aquatic frogs.


title: Structural proteins of equine arteritis virus.

text: We have recently shown that the genome of equine arteritis virus (EAV) contains seven open reading frames (ORFs). We now present data on the structural proteins of EAV and the assignment of their respective genes. Virions are composed of a 14-kDa nucleocapsid protein (N) and three membrane proteins designated M, GS, and GL. M is an unglycosylated protein of 16 kDa, and GS and GL are N-glycosylated proteins of 25 and 30 to 42 kDa, respectively. The broad size distribution of GL results from heterogeneous N-acetyllactosamine addition since it is susceptible to digestion by endo-beta-galactosidase. Using monospecific antisera as well as an antivirion serum, and by expression of individual ORFs, the genes for the structural proteins were identified: ORF 7 codes for N, ORF 6 for M, ORF 5 for GL, and ORF 2 for GS. With the exception of GS, the proteins are about equally abundant in EAV virions, being present at a molar ratio of 3 (N):2 (M):3 (GL). The GS protein, which is expressed at a level similar to that of M in infected cells, is strikingly underrepresented in virus particles (1 to 2%). Our data justify a distinct taxonomic position for EAV, together with lactate dehydrogenase-elevating virus and simian hemorrhagic fever virus; although coronavirus- and toroviruslike in features of transcription and translation, the virion architecture of EAV is fundamentally different.


title: Enterotoxigenic Escherichia coli (ETEC) in farm animals.

text: Animal diseases due to enterotoxigenic Escherichia coli (ETEC) typically appear as severe watery diarrhoea during the first few days of life (also a few days after weaning in pigs). ETEC adhere to the small intestinal microvilli without inducing morphological lesions and produce enterotoxins acting locally on enterocytes. This action results in the hypersecretion (of water and electrolytes) and reduced absorption. Adhesins and toxins are the two prominent virulence attributes of ETEC and the level of knowledge of these factors determines the chances for successful prevention and therapy of the disease. For animal ETEC the most common adhesins are the fimbriae (pili) on the surface: F4(K88), F5(K99), F6(987P), F41, F42, F165, F17 and F18. Enterotoxins (extracellular proteins or peptides) of animal ETEC are classified as heat-labile (LT) and heat-stable (ST) enterotoxins with further subdivisions (LTh-I, LTp-I, LTIIa, LTIIb, STaH, STaP, STb) according to antigenic and biological differences. Fimbriae and LT enterotoxins are made up of large molecular weight proteins which facilitate their utilisation in vaccines and their detection using immunodiagnostic systems. The adhesive fimbriae and enterotoxins of animal ETEC are plasmid determined (except F41 and F17). Virulence gene probes (DNA hybridisation, PCR) are specific and sensitive diagnostic and epidemiologic tools for the detection of ETEC. Based on genetic typing, the ETEC, in spite of limited serogroups, seem to represent a population of E. coli with a diverse genetic background.


title: [Use of multiplex PCR method with real-time detection for differential diagnosis of respiratory viral infections].

text: Multiplex real-time polymerase chain reaction test-system with fluorescent detection (RT-PCR) for simultaneous identification of main agents of acute respiratory viral infections: influenza A (IAV) and B viruses (IBV), parainfluenza viruses types 1, 2, 3, 4 (PIV 1 - 4), adenoviruses (ADV), respiratory syncitial virus (RSV), rhinoviruses (RV) and enteroviruses (EV), in presence internal positive control (IPC) represented by vaccine strain of rubella virus RA 27/3. Using multiplex RT-PCR method, respiratory viruses were detected in 116 out of 226 clinical samples (nasal swabs) obtained from patients with symptoms of acute respiratory infection: in 68 (58.6%) samples--IBV; in 21 (18.1%)--IAV; in 12 (10.3%) --RV; in 6 (5.2%)--PIV 2; in 4--(3.4%) ADV; in 3 (2.6%)--RSV; in 2 (1.7%)--EV; in 2 (1.7%)--PIV 4; in 1 (0.9%)--PIV 3; in 1 (0.9%)--PIV 1. Mixed infection was observed in 4 (3.4%) patients. PCR assay allowed to reveal various respiratory viruses in 51.3% of samples. At the same time samples were tested for the presence of 12 respiratory viruses--IAV, IBV, PIV 1 - 4, RSV, RV, metapneumoviruses, and coronaviruses NL63, 229E and OC43--in the presence of IPC represented by equine arteritis virus using analogous PCR test-system provided by medical center of Leiden university. Results of tests for detection of IAV, IBV, RSV, PIV 1 - 4, and RV, analyzed by both systems, agreed in 94%. Multiplex format of RT-PCR performing significantly reduces time and cost of the test, which make it suitable and effective instrument of epidemiological studies.


title: Immunogenetics of severe respiratory infections: models for the development of new therapeutic strategies.

text: Innate and adaptive immunity plays a critical role in the defence of the lung and other mucosal surfaces exposed to micro-organisms. Anti-microbial peptides and proteins, cytokines and chemokines are important immune weapons as they build up the protective front for the respiratory tract. The notion that susceptibility to infectious diseases may be inherited is widely accepted and, as it is the failure to activate adaptive immunity that may allow infection to become established and progress toward invasion and dissemination, the recognition of specific gene defects affecting the ability of the immune system to overcome invading pathogens may shed light upon those mechanisms of immune regulation that are playing the most critical roles. The aim of the present review is to discuss some of the advances in infection immunogenetics that may lead to identify new strategies in the development of new anti-infectious and anti-inflammatory drugs.


title: [Isolation and identification of SARS virus in Guangdong province].

text: BACKGROUND To isolate and identify pathogen of atypical pneumonia in Guangdong. METHODS Pathogens were isolated from variety of samples collected from atypical pneumonia patient by using MDCK cells, and identified with serological and molecular methods. RESULTS A novel coronavirus was isolated from patients with atypical pneumonia, from which an RNA fragment of 279 nt was amplified by nested RT-PCR. And sequence assay showed that only 39-65 percent of sequence of the virus was homogenous to known coronavirus, but almost 100% homogenous (with one base exception, 12a to t) to SARS-associated coronavirus isolated from patients outside Guangdong, such as in Beijing, Hong Kong, Taiwan, Germany, Italy and so on. Indirect immunofluorescence test showed a specific antigen-antibody reactivity between the coronavirus and convalescent-phase sera of SARS patients. CONCLUSION The pathogen of the atypical pneumonia in Guangdong province was a novel type of coronavirus, which could be isolated by using MDCK cells.


title: Necrotizing enterocolitis in full-term newborns.

text: During a 17-month period, 19 full-term newborn infants were hospitalized for necrotizing enterocolitis (NEC). 10 of these infants had no apparent risk factors. Only 7 suffered acute fetal distress. 16 of the 19 infants were born in the same hospital. Toxicological and bacteriological investigations of possible cause revealed no specific agent. Virological analysis of 8 stool samples revealed the presence of corona virus in 5 of the 8.


Ailing Russian health-care system in urgent need of reform.


title: [Study on the myocardiac injury in patients with severe acute respiratory syndrome].

text: OBJECTIVE To explore the myocardiac injury in patients with severe acute respiratory syndrome (SARS) and its clinical significance. METHODS 37 SARS patients fulfilled the Guangdong provincial diagnostic criteria for infectious atypical pneumonia and 35 health controls were investigated. The serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate transaminase (AST), troponin I (TnI), creatine kinase-MB (CK-MB) and myoglobin (MYO) were measured. RESULTS CK, LDH and AST levels in patients were higher than those of control group (P < 0.01); furthermore, among patients the levels were higher in fatal cases than in survivors. The positive rates of TnI, CK-MB and MYO in patients were higher than those in controls (P < 0.05). CONCLUSION The patients with SARS are subject to complicating myocardiac injury. Therefore, careful monitoring of the myocardiac enzyme profiles is of great importance in reducing the complications and mortality in patients with SARS.


title: Expressing short hairpin RNAs in vivo.

text: Promoter-based expression of short hairpin RNAs (shRNAs) may in principle provide stable silencing of genes in any tissue. As for all approaches that require transgene expression, safe delivery is the biggest obstacle, but toxicity can also occur via expression of the sequence itself. Innate immunity mechanisms can be triggered by expressed hairpin RNAs, critical cellular factors can be saturated, and genes other than the intended target can be silenced. Nevertheless, shRNAs constitute a valuable tool for in vivo research and have great therapeutic potential if the challenges with delivery and side effects are appropriately addressed.


title: Primary care reform in the Peoples' Republic of China: implications for training family physicians for the world's largest country.

text: Just as China has struggled with bringing an expanding economy to a country with large areas of economic underdevelopment, the Chinese health care system is experiencing tension between tradition and modernization. Because of this tension, health care in the Peoples' Republic of China has been undergoing significant reform since the beginning of the 1980s. Experiments in market-based health systems have been unsuccessful and have exacerbated disparities. New reforms, announced in 2006, stress the role of family physicians in leading the health care system. This paper discusses the history of the developments that led up to the new reforms and the educational challenges of training sufficient numbers of family physicians to meet the requirements of the new system.


title: [Molecular detection assays for 2012 identified novel human coronavirus (HCoV) and probe modification with locked nucleic acid (LNA)].

text: OBJECTIVE To develop and optimize the molecular detection assays for recently identified human coronavirus (HCoV) infection. METHODS Based on the 208 base pair(bp) sequence of novel HCoV reported by HPA of UK, we designed and obtained several pairs of primer (F-1, R-1; F-2, R-2) and Taqman probes (TZ1,TZ2) for detection of novel HCoV. Two of probes were modified with LNA (LNA-TZ1, LNA-TZ2). Then, RT-PCR and various real time RT-PCR assays were developed and optimized in this study. We also compared our assays with the real time RT-PCR assays reported recently by Europe team based on upE or ORF1b target. RESULTS The RT-PCR or real time RT-PCR assays for novel HCoV were developed without cross-reactivity with other HCoV and several common respiratory viruses using clinical specimen panel. The analytical sensitivity of assays were less than 50-500 copies per reaction and the detection was improved when Taqman probe modified with LNA-tagged, compared to no LNA-tagged in real time RT-PCR assays. The upE and LNA-TZ1 based assays were better than others. CONCLUSION The molecular detection sensitivity and specificity of TaqMan-based real time PCR assay could be improved when probe tagged with LNA. The upE or LNA-TZ1 based real time RT-PCR assay was recommend for detection of novel HCoV. This study laid a foundation for improving the performance of novel HCoV detection.


Investigation of bovine viral diarrhoea virus-associated abortion in cattle vaccinated with inactivated vaccine.


Reasons for abortion.


title: Intracellular targeting signals contribute to localization of coronavirus spike proteins near the virus assembly site.

text: Coronavirus budding at the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) requires accumulation of the viral envelope proteins at this point in the secretory pathway. Here we demonstrate that the spike (S) protein from the group 3 coronavirus infectious bronchitis virus (IBV) contains a canonical dilysine endoplasmic reticulum retrieval signal (-KKXX-COOH) in its cytoplasmic tail. This signal can retain a chimeric reporter protein in the ERGIC and when mutated allows transport of the full-length S protein as well as the chimera to the plasma membrane. Interestingly, the IBV S protein also contains a tyrosine-based endocytosis signal in its cytoplasmic tail, suggesting that any S protein that escapes the ERGIC will be rapidly endocytosed when it reaches the plasma membrane. We also identified a novel dibasic motif (-KXHXX-COOH) in the cytoplasmic tails of S proteins from group 1 coronaviruses and from the newly identified coronavirus implicated in severe acute respiratory syndrome. This dibasic motif also retained a reporter protein in the ERGIC, similar to the dilysine motif in IBV S. The cytoplasmic tails of S proteins from group 2 coronaviruses lack an intracellular localization signal. The inherent differences in S-protein trafficking could point to interesting variations in pathogenesis of coronaviruses, since increased levels of surface S protein could promote syncytium formation and direct cell-to-cell spread of the infection.


title: Flexor tendon repair with barbed suture: an experimental study.

text: Various suture materials have been used for tendon repair. Barbed suture in particular has been demonstrated to be effective for both wound closure and tendon repair. Ten fresh-frozen flexor digitorum profundus tendons of pig were transected and repaired using the two-strand modified Kirchmayr-Kessler technique. The samples were divided into two groups: 4-0 barbed absorbable polyglyconate sutures and 4-0 monofilament absorbable polyglyconate sutures. We measured tensile strength and distance that the testing system pulled the tendon using a universal testing machine. Tensile strength at which 1 and 2 mm of gapping occurred was measured visually at the repair site. Tendons repaired by barbed sutures showed greater tensile strength than monofilament sutures forming still 2-mm gap. However, statistical significance was obtained only at 1-mm gap formation. We concluded that for tendon repair, barbed sutures were superior material to monofilament sutures because of their greater tensile strength, reducing both the initial tendon load and the risk of dehiscence.


1778: PNEUMOCYSTIS PNEUMONIA REMAINS A DIAGNOSTIC CHALLENGE WITH PCR IN ACUTE SETTINGS.


title: 18th International Conference on Antiviral Research.

text: The 18th International Conference on Antiviral Research (ICAR) was held at the Princess Sofia Hotel in Barcelona, Spain, from 11th-14th April, 2005. This is a yearly international meeting sponsored by the International Society for Antiviral Research (ISAR). The current president of ISAR is John A Secrest 3rd of the Southern Research Institute. The scientific programme committee was chaired by John C Drach from the University of Michigan. ISAR was founded in 1987 to exchange prepublication basic, applied and clinical information on the development of antiviral, chemical and biological agents as well as to promote collaborative research. The ISAR has had a major role in the significant advances of the past decade in the reduction of the societal burdens of viral diseases by the focus of ICAR on the discovery and clinical application of antiviral agents. The 18th ICAR was organised as a series of focus presentations on specific viral groups consisting of oral and poster presentations of original research findings. In addition, the conference included plenary speakers, award presentations, a minisymposium on bioterrorism, and a satellite symposium on clinical antiviral drug developments. The size of the conference (> 50 oral and 250 poster presentations) necessitates limitation to the most noteworthy in the judgment of this reviewer. The current membership of the ISAR is approximately 700 with approximately 50% the membership in attendance.


title: Severe acute respiratory syndrome epidemic in Taiwan, 2003.

text: In Taiwan, since the first case of severe acute respiratory syndrome (SARS) was identified on February 25, 2003, a total of 3032 cases of suspected or probable SARS were reported prior to July 5, 2003. Among these cases, 664 cases were classified as probable SARS based on the clinical case definitions and 346 had a positive result for the SARS-associated coronavirus (SARS-CoV). The epidemic in Taiwan could be divided into 2 distinct stages. In stage I (late-February to mid-April) patients had traceable contact with infected patients or travel histories to known affected areas of SARS. By contrast, patients in stage II (mid-April to June) acquired infection via intra-hospital or inter-hospital transmission. The mortality rate directly attributable to SARS during the 2 stages of the outbreak in Taiwan was 11% (37 patients). Phylogenetic analysis of sequences of SARS-CoV strains in Taiwan and other countries showed that Taiwanese strains were closely related to those isolated from patients in Hong Kong and Guangdong. The nonspecific initial symptoms and signs of the illness, the absence of reliable diagnostic tests, as well as the initial lack of strict infection control measures in hospitals and effective national control policies contributed to the island-wide spread of the SARS epidemic in Taiwan. Development of an effective strategy to prepare for future outbreaks will require the implementation of an active coordinated clinical reporting system, international collaboration to identify cases in the early stage, development of laboratory tools for early diagnosis, a robust system of prepared isolation, and adequate quarantine facilities.


title: Update on pediatric vasculitis.

text: PURPOSE OF REVIEW The purpose of this review is to provide an overview of the recent research and progress made in the field of pediatric vasculitis. RECENT FINDINGS Over the past year, researchers have described several polymorphisms in Henoch-Schonlein purpura and Kawasaki Disease as well as the association between various vasculitides and infections. International and multidisciplinary efforts in Kawasaki Disease have resulted in recommendations for its diagnosis, treatment, and long-term management to improve patient care and further increase global collaboration. Researchers are investigating the role of inflammation and its role in endothelial health and atherosclerosis. Treatment regimens continue to improve, with the use of different immunosuppressive medications; however, we continue to have incomplete treatment successes. SUMMARY Vasculitides are rare conditions with significant morbidity and mortality whose prognosis has improved with newer diagnostic modalities and treatments; however, we continue to have insufficient knowledge of vasculitides and lack unambiguous diagnostic criteria. As technology continues to progress it is clear that a single cause of these diseases may be an oversimplification: the genetic makeup of individuals, as well as various environmental exposures, are of vital importance in the pathophysiology and evolution of disease processes, as well as response to therapy. Efforts should continue to improve international multicenter collaboration and interdisciplinary efforts to help solve this ever-growing puzzle.


title: On the spread of the novel influenza A (H1N1) virus in Mexico.

text: A novel influenza A H1N1 virus of swine origin is responsible for the influenza epidemic affecting Mexico, the United States of America (USA), and 39 other countries. While the origin of this emerging pathogen remains uncertain, an increase in the reported incidence of respiratory diseases was noted during March 2009 at the town of La Gloria, in the southeastern state of Veracruz, Mexico. So far, this is the first community in which a case of novel influenza A H1N1 virus has been identified. Further cases were rapidly detected in other areas of Mexico and elsewhere. Initially, the atypical respiratory disease outbreak caused great uncertainty posing a challenge to the Mexican health system. Control measures such as social distancing, timely medical care, and personal hygiene have so far proven effective in containing the outbreak, resulting in a decline of the number of new cases. To the best of our knowledge, it appears that the virus might not be as virulent or contagious as previously thought. Here we provide a description of the influenza epidemic spread in Mexico. As the virus disseminates worldwide, there is concern about the possibility of a new reassortment resulting in a more pathogenic strain that will pose a threat for every country. The influenza epidemic provided lessons that underscore the importance of epidemiologic surveillance and preparedness. Further investigation to address questions about this new virus and conditions for its spread is warranted.


title: Good governance in 'one health' approaches.

text: The authors discuss 'One Health' approaches for controlling newly recognised and re-emerging diseases of animal origin and contributions towards pandemic preparedness based on enhanced collaboration between Veterinary Services, Human Health Services and Environmental Services. Improved veterinary governance and cooperation with public health managers, social scientists, ecologists and many other stakeholders are important for reducing the risks of potential zoonoses--including foodborne diseases--at their source. Two case studies are presented to illustrate how One Health approaches can make a difference--Hendra disease incidents in Australia and rabies management on the African continent. This article also includes an overview of collaboration at the international level between the Food and Agriculture Organization of the United Natiorfs, the World Organisation for Animal Health, and the World Health Organization. Environmental determinants for disease emergence, anthropogenic climate change and human encroachment on shrinking wildlife habitats are considered, using highly pathogenic avian influenza A (H5N1) and Nipah virus as examples. Finally, the authors discuss the effects of livestock production on environmental change--in the light of global population growth and increasing demand for livestock and aquaculture products--with the need for future policy decisions to be based on a multidisciplinary One Health approach.


title: Computational fluid dynamics for turbomachinery internal air systems.

text: Considerable progress in development and application of computational fluid dynamics (CFD) for aeroengine internal flow systems has been made in recent years. CFD is regularly used in industry for assessment of air systems, and the performance of CFD for basic axisymmetric rotor/rotor and stator/rotor disc cavities with radial throughflow is largely understood and documented. Incorporation of three-dimensional geometrical features and calculation of unsteady flows are becoming commonplace. Automation of CFD, coupling with thermal models of the solid components, and extension of CFD models to include both air system and main gas path flows are current areas of development. CFD is also being used as a research tool to investigate a number of flow phenomena that are not yet fully understood. These include buoyancy-affected flows in rotating cavities, rim seal flows and mixed air/oil flows. Large eddy simulation has shown considerable promise for the buoyancy-driven flows and its use for air system flows is expected to expand in the future.


title: Coronavirus multiplication strategy. I. Identification and characterization of virus-specified RNA.

text: We examined the synthesis of intracellular RNA in primary chicken embryo kidney cells infected with the avian coronavirus infectious bronchitis virus. Infected cells were labeled with (32)P(i) in the presence of actinomycin D for the duration of the viral multiplication cycle, and nucleic acids were extracted, denatured, and analyzed on agarose slab gels. Six major RNA species were found. None of these RNAs was found in extracts of mock-infected cells. All six of the virus-specified RNAs (designated species A through F) were single stranded, and RNA species F had the same electrophoretic mobility as purified viral genome RNA. The molecular weights of the five subgenomic RNAs were estimated to be 0.8 x 10(6), 0.9 x 10(6), 1.3 x 10(6), 1.5 x 10(6), and 2.6 x 10(6) for species A through E, respectively. All of the RNAs were polyadenylated and are therefore likely to be viral mRNA's. The RNAs were synthesized in approximately constant proportions throughout the viral multiplication cycle. Intracellular RNA species A, B, C, D, and F and the purified viral genome were analyzed by RNase T(1) fingerprinting. The results confirmed the identification of RNA species F as the intracellular genome and the derivation of the four smaller RNAs from the genome. Fingerprinting also showed that the intracellular RNAs constitute a nested set such that the nucleotide sequence of each RNA is contained within all larger RNAs and each larger RNA contains an additional sequence congruent with its greater size. Finally, the possible modes of transcription and translation of the infectious bronchitis virus RNAs are discussed.


title: Mechanisms and experimental models of chronic obstructive pulmonary disease exacerbations.

text: Exacerbations of chronic obstructive pulmonary disease (COPD) are a major cause of morbidity, mortality, and rising health care costs. In addition, they are associated with an accelerated loss of lung function and thus have a direct effect on disease progression. There are few studies examining the cellular and molecular mechanisms of COPD exacerbations. Exacerbations are linked to increased airway inflammation and oxidative stress, but many questions remain unanswered regarding the key inflammatory cells and mediators. Current therapies for COPD exacerbations are of limited effectiveness, and a better understanding of the inflammatory events at exacerbation is required to devise new therapeutic agents. The development of experimental models of exacerbation-for example, the use of experimental rhinovirus infection in humans with COPD-would greatly facilitate studies of exacerbations.


title: Airborne inflammatory factors: "from the nose to the brain".

text: The intranasal pathway is a direct route of communication between the environment and the brain. This pathway is currently used for the delivery of several experimental therapeutic peptides and vaccines because it bypasses the blood brain barrier. It is also a route of entrance to the brain for several viruses and toxic substances. Airborne infectious, allergic and pollution agents are among the most common inflammatory factors which may affect brain function via the brain-nose interface. The inflammatory processes triggered in the upper respiratory tract by these agents are positioned to influence the immune response of the brain and therefore, influence its function and alter behavior. Several clinical and epidemiological studies find an association between inflammatory factors affecting the intranasal pathway and neurological disorders such as multiple sclerosis, Alzheimer and Parkinson diseases as well as mental disorders including anxiety and mood disorders. However the mechanisms of interaction between the immune response in the nasal epithelium and the brain are poorly understood. This article discusses current evidence about these mechanisms and associations with neurological and mental diseases.


title: Microarray technology as a universal tool for high-throughput analysis of biological systems.

text: Over the last years microarray technology has become one of the principal platform technologies for the high-throughput analysis of biological systems. Starting with the construction of first DNA microarrays in the 1990s, microarray technology has flourished in the last years and many different new formats have been developed. Peptide and protein microarrays are now applied for the elucidation of interaction partners, modification sites and enzyme substrates. Antibody microarrays are envisaged to be of high importance for the high-throughput determination of protein abundances in translational profiling approaches. First cell microarrays have been constructed to transform microarray technology from an in vitro technology to an in vivo functional analysis tool. All of these approaches share a common prerequisite: the solid support on which they are generated. The demands on this solid support are thereby as manifold as the applications themselves. This review is aimed to display the recent developments in surface chemistry and derivatization, and to summarize the latest developments in the different application areas of microarray technology.


title: [Surgical aspects of endoscopic treatment of sagittal craniosynostosis (scaphocephaly) in children].

text: The article is devoted to the surgical treatment of sagittal craniosynostosis in children. Among non-syndromic monosynostosis sagittal craniosynostosis (scaphocephaly) is the most common. Treatment of children with craniosynostosis should begin as soon as possible. Endoscopic method refers to minimally invasive technique in surgical correction of craniosynostosis. This article presents a features of surgical treatment at all stages of the endoscopic cranioplasty. Presented data is based on the experience of treatment of 20 children with primary sagittal craniosynostosis. Treatment was performed using endoscopic techniques, special tools designed specifically for the endoscopic cranioplasty.


title: Validation of a questionnaire about the perception of occupational biohazard in Spanish companies.

text: OBJECTIVES The aim of this work was to develop, validate and test a new questionnaire to assess the biological risk in workers with intentional or unintentional exposure to biological agents. METHODS A questionnaire including 34 questions was developed to study the perception of workers against occupational biohazard. Content Validity Index (CVI) and Content Validity Ratio (CVR) were calculated for the analysis of content validity. A pilot study was carried out with 60 workers from 17 companies performing analysis of Cronbach's α to assess the internal consistency or reliability. RESULTS A total of 518 workers from 51 Spanish companies in which there is exposure to biological agents participated in the study yielding a response rate of 90%. The final questionnaire obtained a Cronbach's α > 0.759 with a stable test-retest result. The questionnaire validation demonstrates that it could be used to evaluate the biological risks and help the prevention of occupational accidents and diseases. DISCUSSION This study has validated the need to evaluate worker's perception against occupational risks, as well as the application of prevention methods and protective equipment. It is a first step towards developing an occupational biohazards assessment method including all the requirements set by the European Health and Safety Strategy 2013-2020.


title: [Human bocavirus: studies in the literature and in Chile].

text: The human bocavirus (HBoV), virus of the Parvoviridae family, discovered by molecular methods in 2005, has been reported in respiratory samples, stool, urine and blood, both in children and adults. Prevalence rates range from 0.8% in fecal samples of individuals with acute diarrhea, up to 19% in respiratory samples and blood. HBoV has been detected in up to 43% of nasopharyngeal samples in asymptomatic children. In Chile, HBoV was detected in 24.2% of nasopharyngeal swabs in children under 5 years of age with respiratory symptoms of which 74% had coinfection with other viruses. In asymptomatic children under 5 years of age, 37.5% of NP samples were positive for HBoV. We discuss the role of HBoV as a causal agent of respiratory and/or enteric disease in light of the high rates of coinfection and asymptomatic infections.


title: The use of alveolar epithelial type I cell-selective markers to investigate lung injury and repair.

text: Alveolar epithelial type I cells cover most of the internal surface area of the lungs. Ultrastructural studies demonstrate that alveolar epithelial type I cell damage is frequently observed in acute and chronic lung diseases. This article discusses the use of cell-selective proteins as markers for the investigation of injury and repair of the alveolar epithelium. The utility of proteins specific to alveolar epithelial type I cells as diagnostic markers of alveolar epithelial injury in acute lung injury is considered, and expression of proteins selective for alveolar epithelial type I cells in lungs following injury and in fibrosis are discussed.


From smallpox to polio and beyond: disease surveillance in India.


title: Post-earthquake injuries treated at a field hospital --- Haiti, 2010.

text: On January 12, 2010, a 7.0-magnitude earthquake struck Haiti, resulting in an estimated 222,570 deaths and 300,000 persons with injuries. The University of Miami Global Institute/Project Medishare (UMGI/PM) established the first field hospital in Port-au-Prince, Haiti, after the earthquake. To characterize injuries and surgical procedures performed by UMGI/PM and assess specialized medical, surgical, and rehabilitation needs, UMGI/PM and CDC conducted a retrospective medical record review of all available inpatient records for the period January 13-May 28, 2010. This report describes the results of that review, which indicated that, during the study period (when a total of 1,369 admissions occurred), injury-related diagnoses were recorded for 581 (42%) admitted patients, of whom 346 (60%) required a surgical procedure. The most common injury diagnoses were fractures/dislocations, wound infections, and head, face, and brain injuries. The most common injury-related surgical procedures were wound debridement/skin grafting, treatment for orthopedic trauma, and surgical amputation. Among patient records with documented injury-related mechanisms, 162 (28%) indicated earthquake-related injuries. Earthquake preparedness planning for densely populated areas in resource-limited settings such as Haiti should account for injury-related medical, surgical, and rehabilitation needs that must be met immediately after the event and during the recovery phase, when altered physical and social environments can contribute to a continued elevated need for inpatient management of injuries.


Using lessons from the past to plan for pandemic flu.


title: Regulation of hepatitis B virus replication by the phosphatidylinositol 3-kinase-akt signal transduction pathway.

text: The phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) signaling pathway is one of the major oncogenic pathways and is activated in many types of human cancers, including hepatocellular carcinoma. It can also be activated by the hepatitis C virus (HCV) nonstructural 5A (NS5A) protein. In the present study, we set out to determine the regulatory effects of this pathway on the replication of hepatitis B virus (HBV). Our results demonstrate that the expression of a constitutively active Akt1 profoundly inhibited HBV RNA transcription and consequently reduced HBV DNA replication in HepG2 cells. This suppression of HBV gene transcription was apparently mediated by the activation of mTOR, as it was abolished by the mTOR inhibitor rapamycin. Moreover, treatment of HBV-expressing HepG2.2.15 cells with inhibitors of PI3K, Akt, and mTOR increased the transcription of 3.5-kb and 2.4-kb viral RNA as well as the replication of HBV DNA. This observation implies that the basal level activation of this pathway in HepG2 cells regulated HBV replication. Consistent with previous reports showing that the HCV NS5A protein could bind to the p85 subunit of PI3K and activate the PI3K-Akt signal transduction pathway, our results showed that expression of this protein could inhibit HBV RNA transcription and reduce HBV DNA replication in HepG2 cells. Taken together, our results suggest that the activation of the PI3K-Akt pathway during liver oncogenesis may be at least partially responsible for the elimination of HBV replication from tumor cells and may also provide an explanation for the observed suppression of HBV replication by HCV coinfection.


title: Clinical impact and diagnosis of human metapneumovirus infection.

text: BACKGROUND Acute respiratory infections are the most common illnesses experienced by people of all ages worldwide. A portion of hitherto unexplained viral respiratory tract illnesses (RTIs) can now be attributed to the human metapneumovirus (hMPV), which was discovered in 2001. Several surveys on the burden of disease of hMPV infection have been conducted in various study groups and with different diagnostic assays. To estimate the impact of hMPV infection in a hospital setting and in the community at large, we reviewed these surveys to establish the burden of disease of hMPV infection. METHODS Published data and our own additional unpublished data on the clinical impact of hMPV infection were reviewed. RESULTS AND CONCLUSION Worldwide, hMPV infections account for at least 5 to 7% of the RTI in hospitalized children, but immunocompromised and elderly individuals are also at risk. In the general community hMPV infections account for at least 3% of patients who visit a general practitioner for RTI. The seasonality of hMPV infections resembles that for respiratory syncytial virus and influenza virus infections, with recurrent epidemics during the winter months. Clinical symptoms and laboratory findings associated with hMPV infection exhibit a spectrum virtually indistinguishable from those associated with respiratory syncytial virus disease. The development of diagnostic assays must take into account the existence of two hMPV serotypes.


Cell death in planktonic, photosynthetic microorganisms.


title: Gastrointestinal parasites of cougars (Felis concolor) in Washington and the first report of Ollulanus tricuspis in a sylvatic felid from North America.

text: Gastrointestinal helminths including two species of cestodes (Taenia omissa and T. ovis krabbei) and three species of nematodes (Toxocara cati, Cylicospirura subequalis and Ollulanus tricuspis) are reported from two free-ranging cougars (Felis concolor) in Washington (USA). Ollulanus tricuspis is reported for the first time from cougars and represents the first occurrence of this parasite in a sylvatic felid from North America.


title: Assessing the genetic component of the susceptibility of mice to viral infections.

text: Laboratory mice often exhibit wide differences in susceptibility when infected experimentally with viruses. Based on such observations, experiments have been designed to investigate the determinism of these differences at the molecular level, and a few genes that play a major role in the innate mechanisms of defence of the species toward viral aggressions have been characterised. For example, the extraordinary resistance of SJL mice to experimental infections with hepatitis virus strain A59 is the consequence of a structural alteration of a cell adhesion molecule which normally binds to the spikes of the virus, allowing its entry into the cells. If the virus cannot bind to the molecule, or if the molecule is absent, epithelial cells of the intestine and liver are not infected and mice are resistant. In the same way, most--not to say all--laboratory strains of mice are susceptible to infections with orthomyxoviruses or flaviviruses because essential molecules, the synthesis of which is normally triggered by interferon, are defective in these mice. Wild mice, by contrast --probably because they are constantly exposed to natural infections--are resistant. Finally, some mouse strains resist experimental infections by the mouse cytomegalovirus 1 (MCMV-1) because, once infected, these mice synthesise a molecule at the surface of infected cells which allows immediate recognition and killing by natural killer (NK) cells. With the exuberant development of mouse genetics and the constant generation of new mutant alleles, it is likely that many more genes with an impact on the phenotype of resistance or susceptibility will be identified in the forthcoming years. These genes are probably numerous, however, and many of them presumably interact with each other and/or have additive effects. This might slow down progress in our understanding of the innate mechanism of defence.


title: Suspensory ligament rupture technique during ovariohysterectomy in small animals.

text: During ovariohysterectomy, suspensory ligament (SL) rupture permits retraction of the ovary and distal ovarian pedicle through a limited ventral midline incision. This allows the surgeon to confirm that the pedicle is securely double ligated and includes no ovarian remnant. For less experienced surgeons, SL rupture is often difficult and daunting because the ligament is buried within the abdominal viscera and must be identified blindly by palpation. Furthermore, in dogs, the ligament must be digitally disrupted, which may cause hemorrhage and serious injury to surrounding structures such as the ovarian pedicle. This article describes step-by-step techniques to disrupt the SL in dogs and cats. We have found that these techniques can be taught easily and successfully to novice surgeons.


Global changes related to travelers' health.


title: A functional -1 ribosomal frameshift signal in the human paraneoplastic Ma3 gene.

text: A bioinformatics approach to finding new cases of -1 frameshifting in the expression of human genes revealed a classical retrovirus-like heptanucleotide shift site followed by a potential structural stimulator in the paraneoplastic antigen Ma3 and Ma5 genes. Analysis of the sequence 3' of the shift site demonstrated that an RNA pseudoknot in Ma3 is important for promoting efficient -1 frame-shifting. Ma3 is a member of a family of six genes in humans whose protein products contain homology to retroviral Gag proteins. The -1 frameshift site and pseudoknot structure are conserved in other mammals, but there are some sequence differences. Although the functions of the Ma genes are unknown, the serious neurological effects of ectopic expression in tumor cells indicate their importance in the brain.


title: Modelling potential responses to severe acute respiratory syndrome in Japan: the role of initial attack size, precaution, and quarantine.

text: BACKGROUND There has been an outbreak of the severe acute respiratory syndrome (SARS) worldwide. With the use of detailed epidemiological data from other countries, this article describes the possible reason for the SARS epidemic not appearing in Japan, and simulates the impact of different control strategies that can break the transmission cycle of SARS associated coronavirus. METHOD Mathematical modelling is used for predicting the epidemiological outcome and simultaneously for evaluating the effect of interventions on SARS. The study estimates the initial attack size that would result in failed invasion. Three different interventions have been incorporated into the public health response policies; precautionary public health measures, isolation of infected people, and quarantine of exposed humans. RESULTS The maximum number of humans newly infected could be roughly estimated on the basis of the initial attack size, using simple formulas. It is seen that the introduction of only a few cases into certain communities would not lead easily to an epidemic. The possible trajectories of SARS epidemic depend on the levels of public health interventions as quarantine and precautionary public health measures greatly affected the transmissibility of the disease. It is shown that there exist threshold levels of interventions at which the SARS epidemic settles down. CONCLUSION Initial attack size is one of the determinants of whether SARS can successfully invade the community or not. Two of the most effective policy procedures to prevent new infections would be to apply stringent precautionary measures and to impose quicker and more effective quarantine of the exposed populace.


title: Battery performance of 4 intensive care ventilator models.

text: BACKGROUND Hospital electrical power failure represents an important challenge in the intensive care unit. Despite the presence of backup generators, total electrical power failure may still occur. Life-support equipment should have a reliable internal battery to ensure patient safety. We tested the duration of operation of the internal battery of 4 intensive care ventilators. METHODS In our laboratory we evaluated one each of 4 ventilator models available in our facility (Evita XL, Puritan Bennett 840, Avea, and Servo 300), with volume-control and pressure-control ventilation, and with positive end-expiratory pressure (PEEP) of zero and 20 cm H(2)O. We then randomly selected and tested 6 Evita XL and 4 Servo 300 ventilators from our inventory to determine the variability of internal battery duration among ventilators of the same model. The ventilator settings were identical to the previous tests, other than fraction of inspired oxygen, which was set at 0.6, and PEEP was 5 cm H(2)O. RESULTS The battery-duration range of the tested ventilators was 20.5-170.5 min, and the mean +/- SD battery duration was 80.4 +/- 49.3 min. Changes in breath type and PEEP did not significantly impact battery duration. Among the ventilators of the same model, the battery-duration range was 5-69 min and the mean +/- SD battery duration was 28.9 +/- 21.4 min. Use of a compressor significantly shortened battery duration. There was no correlation between battery duration and battery age (r = -0.263). CONCLUSIONS The duration of ventilator operation on internal battery ranged widely among the tested devices. Clinicians need to be aware of these differences in the event of power failure.


title: Triple decoding of hepatitis C virus RNA by programmed translational frameshifting.

text: Ribosomes can be programmed to shift from one reading frame to another during translation. Hepatitis C virus (HCV) uses such a mechanism to produce F protein from the -2/+1 reading frame. We now report that the HCV frameshift signal can mediate the synthesis of the core protein of the zero frame, the F protein of the -2/+1 frame, and a 1.5-kDa protein of the -1/+2 frame. This triple decoding function does not require sequences flanking the frameshift signal and is apparently independent of membranes and the synthesis of the HCV polyprotein. Two consensus -1 frameshift sequences in the HCV type 1 frameshift signal facilitate ribosomal frameshifts into both overlapping reading frames. A sequence which is located immediately downstream of the frameshift signal and has the potential to form a double stem-loop structure can significantly enhance translational frameshifting in the presence of the peptidyl-transferase inhibitor puromycin. Based on these results, a model is proposed to explain the triple decoding activities of the HCV ribosomal frameshift signal.