title: A cross-sectional study of primary-care physicians in Singapore on their concerns and preparedness for an avian influenza outbreak.

text: INTRODUCTION During an avian influenza (AI) pandemic, primary-care physicians (PCPs) are expected to play key roles in the prevention and control of the disease. Different groups of PCPs could have different concerns and preparedness level. We assessed the concerns, perceived impact and preparedness for an outbreak among PCPs in Singapore. MATERIALS AND METHODS A cross-sectional survey of PCPs working in private practice (n=200) and public clinics (n=205) from March to June 2006 with an anonymous self-administered questionnaire on concerns (12- items), perceived impact (10 items) and preparedness (10 items) for an outbreak. RESULTS Two hundred and eighty-five PCPs responded - 149 (response rate: 72.7%) public and 136 (response rate: 67.3%) private. The majority were concerned about risk to their health from their occupation (95.0%) and falling ill with AI (89.7%). Most (82.5%) accepted the risk and only 33 (11.8%) would consider stopping work. For perceived impact, most felt that people would avoid them (69.6%) and their families (54.1%). The majority (81.3%) expected an increased workload and feeling more stressed at work (86.9%). For preparedness, 78.7% felt personally prepared for an outbreak. Public PCPs were more likely to be involved in infection-control activities and felt that their workplaces were prepared. CONCLUSIONS Most PCPs felt personally prepared for an outbreak but were concerned about their exposure to AI and falling ill. Other concerns included social ostracism for themselves and their families. Public PCPs appeared to have a higher level of preparation. Addressing concerns and improving level of preparedness are crucial to strengthen the primary-care response for any AI outbreak.


title: Nucleotide sequence of the glycoprotein S gene of bovine enteric coronavirus and comparison with the S proteins of two mouse hepatitis virus strains.

text: The gene encoding the spike glycoprotein (S) of bovine enteric coronavirus (BECV) was cloned and its complete sequence of 4092 nucleotides was determined. This sequence contained a single long open reading frame with a coding capacity of 1363 amino acids (Mr 150,747). The predicted protein had 19 N-glycosylation sites. A signal sequence comprising 17 amino acids was observed starting from the first methionine residue. A potential peptidase cleavage site was located between amino acids 763 and 767. These cleavage explain the maturation of the primary product of the S gene to S1 (Mr 104,692) and S2 (Mr 84,175) spike structural proteins. Two amphipathic alpha-helices (amino acids 1007 to 1077 and 1269 to 1294) which may constitute the 12 nm stalk of the viral spike were also observed; another alpha-helix (amino acids 1305 to 1335) may be involved in the anchorage of the spike in the viral membrane. Comparison of this protein sequence to the described homologous mouse hepatitis (MHV) strain A59 and MHV-JHM S protein sequences led us to suggest that MHV-A59 and MHV-JHM S genes could be derived from a deletion of the BECV S gene.


title: Antigenic and genomic relatedness of turkey-origin coronaviruses, bovine coronaviruses, and infectious bronchitis virus of chickens.

text: In earlier studies in our laboratory, we found that bovine coronavirus (BCV) was pathogenic for 1-day-old turkey poults. This finding prompted us to study the antigenic and genomic relatedness of turkey origin coronaviruses (TOCVs) to BCV. A one-step reverse transcription (RT)-polymerase chain reaction (PCR) targeting a 730-base pair fragment of the nucleocapsid (N) gene of BCV and a nested PCR targeting a 407-base pair fragment of the N gene were used in an attempt to detect TOCV from North Carolina, Indiana, and a prototype turkey coronavirus (TCV) obtained from the American Type Culture Collection. Both the one-step RT-PCR and the nested PCR amplified cell culture-passaged isolates of calf diarrhea strains of BCV but none of the 15 tested TOCVs or transmissible gastroenteritis coronavirus of swine. TOCVs also did not cross-react in a BCV antigen-capture (AC) enzyme-linked immunosorbent assay (ELISA) system with monoclonal antibodies (MAbs) against N, spike glycoprotein, and hemagglutinin esterase glycoprotein proteins of BCV as coating antibodies. The same TOCVs could be detected with primers designed from the genome of infectious bronchitis virus (IBV) of chickens. These primers amplified a 1082-base pair region spanning portions of the membrane glycoprotein (M) and N protein genes of IBV and TCV. The TOCVs also cross-reacted in an AC-ELISA with MAbs against the M and subunit 2 of spike glycoprotein of IBV.


title: Antibody-dependent enhancement of hepatitis C virus infection.

text: Hepatitis C virus (HCV) often causes a persistent infection associated with hypergammaglobulinemia, high levels of antiviral antibody and circulating immune complexes, and immune complex disease. We previously reported that only a limited neutralizing activity to vesicular stomatitis virus or HCV pseudotype is generated in animals immunized with recombinant HCV envelope proteins and chronically infected HCV patient sera. Interestingly, when some of these neutralizing sera were diluted into a range of concentrations below those that reduced virus plaque number, an increase in pseudotype plaque formation was observed. Purified HCV E2-specific human monoclonal antibodies were used to further verify the specificity of this enhancement, and one- to twofold increases were apparent on permissive Huh-7 cells. The enhancement of HCV pseudotype titer could be inhibited by the addition of a Fc-specific anti-human immunoglobulin G Fab fragment to the virus-antibody mixture prior to infection. Treatment of cells with antibody to Fc receptor I (FcRI) or FcRII, but not FcRIII, also led to an inhibition of pseudotype titer enhancement in an additive manner. Human lymphoblastoid cell line (Raji), a poor host for HCV pseudotype infection, exhibited a four- to sixfold enhancement of pseudotype-mediated cell death upon incubation with antibody at nonneutralizing concentrations. A similar enhancement of cell culture-grown HCV infectivity by a human monoclonal antibody was also observed. Taken together, antibodies to viral epitopes enhancing HCV infection need to be taken into consideration for pathogenesis and in the development of an effective vaccine.


title: Critically ill patients with COVID-19 in Hong Kong: a multicentre retrospective observational cohort study

text: Objective To report the first eight cases of critically ill patients with coronavirus disease 2019 (COVID-19) in Hong Kong, describing the treatments and supportive care they received and their 28-day outcomes. Design Multicentre retrospective observational cohort study. Setting Three multidisciplinary intensive care units (ICUs) in Hong Kong. Participants All adult critically ill patients with confirmed COVID-19 admitted to ICUs in Hong Kong between 22 January and 11 February 2020. Main outcome measure 28-day mortality. Results Eight out of 49 patients with COVID-19 (16%) were admitted to Hong Kong ICUs during the study period. The median age was 64.5 years (range, 42–70) with a median admission Sequential Organ Failure Assessment (SOFA) score of 6 (IQR, 4–7). Six patients (75%) required mechanical ventilation, six patients (75%) required vasopressors and two (25%) required renal replacement therapy. None of the patients required prone ventilation, nitric oxide or extracorporeal membrane oxygenation. The median times to shock reversal and extubation were 9 and 11 days respectively. At 28 days, one patient (12%) had died and the remaining seven (88%) all survived to ICU discharge. Only one of the survivors (14%) still required oxygen at 28 days. Conclusion Critically ill patients with COVID-19 often require a moderate duration of mechanical ventilation and vasopressor support. Most of these patients recover and survive to ICU discharge with supportive care using lung protective ventilation strategies, avoiding excess fluids, screening and treating bacterial co-infection, and timely intubation. Lower rather than upper respiratory tract viral burden correlates with clinical severity of illness.


title: [Community-acquired pneumonia].

text: Pneumonias are inflammatory diseases of the lung parenchyma, infection is one of possible causes. With regard to the causes of community acquired pneumonias it is possible to distinguish those typical ones (caused by pneumococcus, legionella and other bacteria) and atypical ones (caused by mycoplasma, chlamydiae and others). Contrary to the atypical ones, typical pneumonias are characterized by sudden onset, high fever, chills, sometimes bloody expectoration and pains, as well as segmental or lobar changes and high leukocyte counts. Patients with tachycardia, diastolic blood pressure below 60 mm Hg and a blood urea nitrogen (BUN) of more than 7 mmol/l, as well as those with chronic basic diseases and a severe course should be hospitalized, further also those, who do not improve after 2 or 3 days therapy, in all cases of suspected pneumonia, with smokers and with patients aged over 40 years, a thorax X-ray should be executed. Typical pneumonias should be treated with penicillin or macrolide antibiotics, atypical ones with macrolide antibiotics, pneumonias with severe course additionally with a second generation cephalosporin. Where these simple rules are observed, a reduction of the still high mortality due to externally acquired pneumonias might be expected.


title: The broad anti-viral agent glycyrrhizin directly modulates the fluidity of plasma membrane and HIV-1 envelope.

text: Cell entry of enveloped viruses requires a wide-fusion-pore mechanism, involving clustering of fusion-activated proteins and fluidization of the plasma membrane and viral envelope. In the present study, GL (glycyrrhizin) is reported to lower membrane fluidity, thus suppressing infection by HIV, influenza A virus and vesicular stomatitis virus, but not by poliovirus. GL-treated HIV-1 particles showed reduced infectivity. GL also inhibited cell-to-cell fusion induced by HIV-1 and HTLV-I (human T-cell leukaemia virus type I). However, when cells treated with 1 mg/ml GL were placed in GL-free medium, they showed increased susceptibility to HIV-1 infection and HTLV-I fusion due to enhancement of membrane fluidity. The membrane dependence of GL and GL removal experiments suggest that GL does affect the cell entry of viruses. HIVs with more gp120 were less dependent on temperature and less sensitive to GL treatment than those with less gp120, indicating that the existence of more gp120 molecules resulted in a higher probability of forming a cluster of fusion-activated proteins.


title: Dissemination of multisusceptible methicillin-resistant Staphylococcus aureus in Singapore.

text: Analysis of hospital-acquired methicillin-resistant Staphylococcus aureus strains isolated from a tertiary public hospital in Singapore revealed that multisusceptible strains had gradually started to replace the endemic multiresistant strain (ST239-MRSA-III) since 2002. Molecular typing showed that this was a predominantly clonal outbreak of a UK-EMRSA-15 strain (ST22-MRSA-IV).


title: Natural killer cell activation after infection with lactate dehydrogenase-elevating virus.

text: Early after infection, lactate dehydrogenase-elevating virus (LDV) alters the immune system by polyclonally activating B lymphocytes, which leads to IgG2a-restricted hypergammaglobulinaemia, and by suppressing the secretion of Th2 cytokines. Considering that these alterations may involve cells of the innate immune system and cytokines such as interferon-gamma (IFN-gamma), we analysed the effect of LDV on natural killer (NK) cells. Within a few days of infection, a strong and transient NK cell activation, characterized by enhanced IFN-gamma message expression and cytolysis, was observed. LDV triggered a large increase in serum IFN-gamma levels. Because NK cells and IFN-gamma may participate in the defence against virus infection, we analysed their possible role in the control of LDV titres with a new agglutination assay. Our results indicate that neither the activation of NK cells nor the IFN-gamma secretion affect the early and rapid virus replication that follows LDV inoculation.


title: [Studying the possibility of respiratory immunization against tick-borne encephalitis].

text: There are known 3 likely mechanisms of virus conveyance into the central nervous system (CNS). These include hematogenic penetration, spread along the peripheral nerves, and the olfactory pathway which begins from the infected olfactory neuroepithelial cells. The possibility of viral spread into CNS via the olfactory pathway was shown for the representatives of togaviruses, herpesviruses, coronaviruses, rhabdoviruses, and for some others. This study suggests that the olfactory pathway of viral conveyance into CNS may be blocked by specific mucosal antibodies in the nasal mucosa. The recombinant TK- variant of WR vaccinia strain with inserted genes coding structural and nonstructural proteins of TBE virus is accumulated in the branches of the respiratory tract only while the parenteral vaccinia strain is detected in the brain regions, spleen, respiratory tract, and in blood. The protective activity of recombinant strain and inactivated TBE vaccine after mice immunization by escarification or intranasally, or subcutaneously was comparatively studied. The findings indicate that intranasal immunization by recombinant strain is the most protective against intraperitoneal challenge by TBE virus. The mucosal and humoral immune response that was induced by intranasal immunization seems to provide the highest levels of protection, which was experimentally observed.


title: Comparison of the replication of distinct strains of human coronavirus OC43 in organotypic human colon cells (Caco-2) and mouse intestine.

text: Three strains of human coronavirus (HCV) OC43 were compared for their ability to cause enteric infections and to induce interferon alpha (IFN alpha) using the Caco-2 human colon carcinoma cell line which exhibits spontaneous epithelial differentiation in vitro. MRC-5 cell culture grown stocks were prepared from: 1. CV Paris, a strain of OC43 recovered from an outbreak of necrotizing enterocolitis in newborns. 2. CV Mb, a neurotropic strain of OC43 which exhibits strict neuronal specificity in murine neuronal cell cultures. 3. CV Rd, a strain of OC43 which grows to a high titer in human rhabdomyosarcoma (RD) cells. Immunofluorescent staining for nucleocapsid antigen and plaque assay in MRC-5 cells was used to detect viral replication. BG-9 (human foreskin) cells challenged with vesicular stomatitis virus were used to detect IFN alpha production by human peripheral blood monocytes (PBMC) stimulated by virus infected Caco-2 cells. Caco-2 cells infected with virus at a multiplicity of infection of 0.5 yielded 10(4.6) and 10(4.4) plaque forming units/ml (pfu/ml) with CV Rd and CV Paris respectively, while CV Mb yielded only 10(3) pfu/ml. Caco-2 cells infected with CV Rd induced 64 IU/ml of IFN alpha in PBMC while these cells infected with CV Paris induced less than 2 IU/ml IFN alpha. In cells infected with CV Mb 4 IU/ml IFN alpha was detected. The results suggest that a lack of IFN alpha induction by CV Paris may be an indicator of its enteropathogenic potential.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Severe acute respiratory syndrome coronavirus 7a accessory protein is a viral structural protein.

text: Severe acute respiratory syndrome coronavirus (SCoV) 7a protein is one of the viral accessory proteins. In expressing cells, 7a protein exhibits a variety of biological activities, including induction of apoptosis, activation of the mitogen-activated protein kinase signaling pathway, inhibition of host protein translation, and suppression of cell growth progression. Analysis of SCoV particles that were purified by either sucrose gradient equilibrium centrifugation or a virus capture assay, in which intact SCoV particles were specifically immunoprecipitated by anti-S protein monoclonal antibody, demonstrated that 7a protein was associated with purified SCoV particles. Coexpression of 7a protein with SCoV S, M, N, and E proteins resulted in production of virus-like particles (VLPs) carrying 7a protein, while 7a protein was not released from cells expressing 7a protein alone. Although interaction between 7a protein and another SCoV accessory protein, 3a, has been reported, 3a protein was dispensable for assembly of 7a protein into VLPs. S protein was not required for the 7a protein incorporation into VLPs, and yet 7a protein interacted with S protein in coexpressing cells. These data established that, in addition to 3a protein, 7a protein was a SCoV accessory protein identified as a SCoV structural protein.


title: Mapping membrane activity in undiscovered peptide sequence space using machine learning.

text: There are some ∼1,100 known antimicrobial peptides (AMPs), which permeabilize microbial membranes but have diverse sequences. Here, we develop a support vector machine (SVM)-based classifier to investigate ⍺-helical AMPs and the interrelated nature of their functional commonality and sequence homology. SVM is used to search the undiscovered peptide sequence space and identify Pareto-optimal candidates that simultaneously maximize the distance σ from the SVM hyperplane (thus maximize its "antimicrobialness") and its ⍺-helicity, but minimize mutational distance to known AMPs. By calibrating SVM machine learning results with killing assays and small-angle X-ray scattering (SAXS), we find that the SVM metric σ correlates not with a peptide's minimum inhibitory concentration (MIC), but rather its ability to generate negative Gaussian membrane curvature. This surprising result provides a topological basis for membrane activity common to AMPs. Moreover, we highlight an important distinction between the maximal recognizability of a sequence to a trained AMP classifier (its ability to generate membrane curvature) and its maximal antimicrobial efficacy. As mutational distances are increased from known AMPs, we find AMP-like sequences that are increasingly difficult for nature to discover via simple mutation. Using the sequence map as a discovery tool, we find a unexpectedly diverse taxonomy of sequences that are just as membrane-active as known AMPs, but with a broad range of primary functions distinct from AMP functions, including endogenous neuropeptides, viral fusion proteins, topogenic peptides, and amyloids. The SVM classifier is useful as a general detector of membrane activity in peptide sequences.


title: Hot-cold empathy gaps and medical decision making.

text: Prior research has shown that people mispredict their own behavior and preferences across affective states. When people are in an affectively "cold" state, they fail to fully appreciate how "hot" states will affect their own preferences and behavior. When in hot states, they underestimate the influence of those states and, as a result, overestimate the stability of their current preferences. The same biases apply interpersonally; for example, people who are not affectively aroused underappreciate the impact of hot states on other people's behavior. After reviewing research documenting such intrapersonal and interpersonal hot-cold empathy gaps, this article examines their consequences for medical, and specifically cancer-related, decision making, showing, for example, that hot-cold empathy gaps can lead healthy persons to expose themselves excessively to health risks and can cause health care providers to undertreat patients for pain.


Long-term cure of disseminated Mycobacterium avium infection in a cat.


Immunomodulatory activities of the herbal formula Kwan Du Bu Fei Dang in healthy subjects: a randomised, double-blind, placebo-controlled study.


title: High prevalence of the CD14-159CC genotype in patients infected with severe acute respiratory syndrome-associated coronavirus.

text: To investigate whether genetic factors of innate immunity might influence susceptibility and/or progression in individuals infected with SARS, we evaluated the CD14 gene polymorphism in 198 Hong Kong blood donors and 152 Hong Kong severe acute respiratory syndrome (SARS) patients who were previously genotyped for FcgammaRIIA polymorphisms. The prevalence of the CD14-159CC polymorphism was significantly higher in the patients with severe SARS than in the those with mild SARS or controls (31% versus 15% [mild SARS] or 20% [controls]; mild SARS: P = 0.029; odds ratio, 2.74; 95% confidence interval, 1.15 to 6.57; controls, P = 0.04; odds ratio, 2.41; 95% confidence interval, 1.05 to 5.54), and both CD14-159CC and FcgammaRIIA-RR131 are risk genotypes for severe SARS-CoV infection.


title: In vivo and in vitro models of demyelinating disease: a phosphoprotein phosphatase in host cell endosomes dephosphorylating the nucleocapsid protein of coronavirus JHM.

text: We have identified a phosphoprotein phosphatase which dephosphorylates efficiently the NC protein of coronavirus JHM. The activity was found in L-2 murine fibroblasts, Wistar Furth rat neonatal brain extracts, Wistar Furth rat oligodendrocyte primary cells and in Roc-1 cells, an oligodendrocytic hybrid cell line. In both L-2 cells and Roc-1 cells the enzyme was found to be localized predominantly in the endosomal fraction. The enzyme is optimally active at pH 7.0 and has a requirement for Mn++ ions. This PPPase activity is distinguishable from acidic and alkaline phosphatases. In view of the specificity of the endosomal PPPase for the phosphory-lated NC protein it is hypothesized that this enzyme may have a function during early stages of coronavirus infection.


title: Comparative study between meshes of polyester with collagen and polytetrafluoroethylene in the repair of defects produced in abdominal wall of rats.

text: PURPOSE To study the intraperitoneal use of polyester with collagen and polytetrafluoroethylene meshes in the correction of total ventral wall defects in rats. METHODS Thirty two rats were evaluated and divided randomly into four groups and underwent laparotomy and preparation of total defects of the abdominal wall. Next, the correction of the defect with the intraperitoneal placement of the chosen mesh was performed. The rats were submitted to euthanasia at 30 and 90 days after surgery. Were analyzed the macroscopic adhesions and microscopic aspects, and applied stress rupture test RESULTS All animals showed intraperitoneal adhesions in varying degrees, with no statistical significance difference. There was no difference also between groups in the evaluation of stress rupture tests. On the microscopic aspect, the A30 group had less inflammatory reaction and less formation of granulomas and foreign body reaction that the B30 group, with significant difference. CONCLUSIONS There was no difference in intraperitoneal adhesion and tensile rupture strength among groups. Group B30 presented granulomatous inflammatory reaction at the site of mesh attachment to the wall significantly higher than the A30.


title: Multiple sclerosis and Kaposi's sarcoma--chronic diseases associated with new human herpesviruses?

text: Two diseases that for many years have been suspected to be of viral origin are multiple sclerosis (MS) and Kaposi's sarcoma (KS). With the use of a new technique called representational difference analysis both these diseases have recently been associated with new lymphotropic herpesviruses, i.e. human herpesviruses (HHV) 6 and 8. HHV-6 is a ubiquitous virus and the etiological agent of exanthema subitum. Viral neuroinvasion occurs frequently in primary HHV-6 infection, and meningitis, encephalitis and demyelination have been described as rare complications. A relation with MS has been suggested for HHV-6, as the virus has been detected in MS plaques in the brain. Data from different studies are, however, conflicting and a definitive role for HHV-6 in MS pathogenesis has not been established. HHV-8 is believed to be the causative agent of KS, and is also associated with some rare hematological malignancies. The viral genome contains several potential oncogenes that are believed to have been picked up from the human genome during evolution. The role of HHV-8 in healthy, immunocompetent individuals is however uncertain. In conclusion, the full spectrum of human diseases associated with these new viruses is not yet understood, and rapid developments in molecular biology will continue to shed new light on the interactions between herpesviruses and their hosts.


title: Review of safety and minimum pricing of nitazoxanide for potential treatment of COVID-19.

text: Background Many treatments are being assessed for repurposing to treat coronavirus disease 2019 (COVID-19). One drug that has shown promising results in vitro is nitazoxanide. Unlike other postulated drugs, nitazoxanide shows a high ratio of maximum plasma concentration (Cmax), after 1 day of 500 mg twice daily (BD), to the concentration required to inhibit 50% replication (EC50) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (Cmax : EC50 roughly equal to 14:1). As such, it is important to investigate the safety of nitazoxanide for further trials. Furthermore, treatments for COVID-19 should be cheap to promote global access, but prices of many drugs are far higher than the costs of production. We aimed to conduct a review of the safety of nitazoxanide for any prior indication and calculate its minimum costs of production. Methods A review of nitazoxanide clinical research was conducted using EMBASE and MEDLINE databases, supplemented by ClinicalTrials.gov. We searched for phase 2 or 3 randomised controlled trials (RCTs) comparing nitazoxanide with placebo or active control for 5-14 days in participants experiencing acute infections of any kind. Data extracted were grade 1-4 and serious adverse events (AEs). Data were also extracted on gastrointestinal (GI) AEs, as well as hepatorenal and cardiovascular effects.Active pharmaceutical ingredient cost data from 2016 to 2019 were extracted from the Panjiva database and adjusted for 5% loss during production, costs of excipients, formulation, a 10% profit margin and tax. Two dosages, at 500 mg BD and a higher dose of 1100 mg three times daily (TDS), were considered. Our estimated costs were compared with publicly available list prices from a selection of countries. Results Nine RCTs of nitazoxanide were identified for inclusion. These RCTs accounted for 1514 participants and an estimated 95.3 person-years-of-follow-up. No significant differences were found in any of the AE endpoints assessed, across all trials or on subgroup analyses of active- or placebo-controlled trials. Mild GI AEs increased with dose. No hepatorenal or cardiovascular concerns were raised, but few appropriate metrics were reported. There were no teratogenic concerns, but the evidence base was very limited.Based on a weighted-mean cost of US $61/kg, a 14-day course of treatment with nitazoxanide 500 mg BD would cost $1.41. The daily cost would therefore be $0.10. The same 14-day course could cost $3944 in US commercial pharmacies, and $3 per course in Pakistan, India and Bangladesh. At a higher dose of 1100 mg TDS, our estimated cost was $4.08 per 14-day course, equivalent to $0.29 per day. Conclusion Nitazoxanide demonstrates a good safety profile at approved doses. However, further evidence is required regarding hepatorenal and cardiovascular effects, as well as teratogenicity. We estimate that it would be possible to manufacture nitazoxanide as generic for $1.41 for a 14-day treatment course at 500 mg BD, up to $4.08 at 1100 mg TDS. Further trials in COVID-19 patients should be initiated. If efficacy against SARS-CoV-2 is demonstrated in clinical studies, nitazoxanide may represent a safe and affordable treatment in the ongoing pandemic.


Cellular receptors for viruses: links to tropism and pathogenesis.


title: Retained and fractured microcatheter: a cause of transient ischaemic attacks: endovascular management using carotid stents.

text: A retained microcatheter is a rare complication of endovascular treatment of cerebral aneurysms. We describe such a case that was complicated by delayed microcatheter fracture within the internal carotid artery and subsequent thrombo-embolism resulting in transient ischaemic attacks. We also describe endovascular management of this complication through the use of several carotid stents.


International Congress of Neurolmmunology Edinburgh, September 3 to 7, 2001.


title: Urgent and Emergent Eye Care Strategies to Protect Against COVID-19.

text: Objective This article presents additional strategies to the medical professional and support tactics to keep both the health care provider and patient as safe as possible during the COVID-19 pandemic. Observations Follicular conjunctivitis has been reported as an early sign of infection or during hospitalization for severe COVID-19 disease. It has been confirmed that COVID-19 is transmitted through both respiratory droplets and direct contact. Another possible route of viral transmission is entry through aerosolized droplets into the tears, which then pass through the nasolacrimal ducts and into the respiratory tract. For nonemergent care, eye care providers should use telehealth. Eye care providers should prioritize patient care in order of absolute necessity, such as sudden vision loss, sudden onset flashes and floaters, and eye trauma. In those cases, exposure should be minimized. The close proximity between eye care providers and their patients during slit-lamp examination may require further precautions, such as shields, barriers, and mask use to lower the risk of transmission via droplets or through hand to eye contact. Conclusions All nonemergent eye care appointments should be delayed or conducted remotely. For emergent in-person appointments, careful and appropriate adherence to Centers for Disease Control and Prevention recommendations may minimize exposure for both the health care provider and patient.


title: Cross-protection against a human enteric coronavirus and a virulent bovine enteric coronavirus in gnotobiotic calves.

text: A group 2 human coronavirus designated HECV-4408 was isolated from a child with acute diarrhea and is antigenically and genetically more closely related to bovine coronavirus (BCoV) than to human coronavirus OC43 (X. M. Zhang, W. Herbst, K. G. Kousoulas, and J. Storz, J. Med. Virol. 44:152-161, 1994). To determine whether HECV-4408 infects gnotobiotic calves and induces cross-protective immunity against the virulent enteric BCoV DB2 strain, gnotobiotic calves (n = 4) were orally inoculated with HECV-4408 and then challenged with BCoV DB2 at postinoculation day (PID) 21. All calves inoculated with HECV-4408 developed diarrhea at PID 3 to 4 lasting 5 to 9 days. Fecal and nasal virus shedding were first detected by reverse transcription-PCR at PID 3 to 4 and at PID 2 to 4, respectively. After challenge with bovine coronavirus, no diarrhea or virus shedding was detected in calves inoculated with HECV-4408, but a mock-inoculated calf developed diarrhea and fecal and nasal shedding. Fecal immunoglobulin A (IgA) and serum IgG antibodies were first detected at PID 7 and PID 14, respectively. At postchallenge day 7, serum IgG and fecal IgA antibody titers remained the same or increased only twofold compared to prechallenge titers. An additional two gnotobiotic calves were inoculated with HECV-4408 and euthanized at PID 5. Moderate villous atrophy was observed in the small intestines, and viral antigen was detected in villous enterocytes of the small and large intestines by immunohistochemistry. These results support and extend the previous report that HECV-4408 is likely a variant of bovine coronavirus. They confirm its infectivity for calves and complete cross-protection against a bovine coronavirus (DB2 strain) showing 98.2% amino acid identity to HECV-4408 in the S protein.


title: Utilization of monoclonal antibodies for antigenic characterization of coronaviruses.

text: Hybrid cells secreting monoclonal antibodies against Bovine Enteric Coronavirus (BECV strain G110) were obtained by fusion between SP2/O myeloma cells and splenocytes of mice hyperimmunized with purified virus. Specificity of 8 from the 12 monoclonal antibodies was established to be anti-GP105 by immunochemical staining of viral polypeptides and by immunoprecipitation of radioactive-labelled viral proteins. The reactivity of three BECV isolates (strain G110, F15 and NCDCV) with the different monoclonal antibodies was studied by indirect immunofluorescence (IIF), ELISA, hemagglutination-inhibition and seroneutralization. G110 and F15 interacted in a similar manner with the monoclonal antibodies, but NCDCV failed to react with A9 monoclonal antibody. IIF test allowed us to detect another difference between U.K. BECV isolate and other BECV, in that the U.K. isolate did not react with F7 monoclonal antibody. No differences could be seen by IIF in the reactivity of Danish BECV isolate, Human Enteric Coronavirus (HECV), Bovine Respiratory Coronavirus (BRCV) and our BECV strains, with the 12 monoclonal antibodies. Human Respiratory Coronavirus (OC43), Porcine Hemagglutinating Encephalitis Virus (HEV) and BECV share common antigenic determinants which are outlined by the 6 non-neutralizing monoclonal antibodies. On the basis of the reactivity of coronaviruses with anti-BECV monoclonal antibodies, a distinction can be easily made between BECV and other coronaviruses, and differences were also detected within the BECV isolates. Interestingly HECV were found to be more closely related to BECV than to human OC43. These results show that monoclonal antibodies are of valuable help for biochemical and antigenic studies as well as for diagnostic procedures.


title: Efficacy of an accelerated hydrogen peroxide disinfectant to inactivate porcine epidemic diarrhea virus in swine feces on metal surfaces.

text: In May of 2013, porcine epidemic diarrhea virus (PEDV) was detected in swine for the first time in North America. It spread rapidly, in part due to contaminated livestock trailers. The objective of this study was to test the efficacy of an accelerated hydrogen peroxide disinfectant for inactivating PEDV in the presence of feces on metal surfaces, such as those found in livestock trailers. Three-week-old barrows were inoculated intragastrically with 5 mL of PEDV-negative feces for the negative control, 5 mL of untreated PEDV-positive feces for the positive control, and 5 mL or 10 mL of PEDV-positive feces that was subjected to treatment with a 1:16 or 1:32 concentrations of accelerated hydrogen peroxide disinfectant for a contact time of 30 min at 20°C. These pigs served as a bioassay to determine the infectivity of virus following treatment. Rectal swabs collected from the inoculated pigs on days 3 and 7 post-inoculation were tested by using PEDV-specific real-time reverse transcription polymerase chain reaction and the proportion of pigs in each group that became infected with PEDV was assessed. None of the pigs used for the bioassay in the 4 treatment groups and the negative control group became infected with PEDV, which was significantly different from the positive control group (P < 0.05) in which all pigs were infected. The results suggest that the application of the accelerated hydrogen peroxide under these conditions was sufficient to inactivate the virus in feces found on metal surfaces.


title: The aetiology of SARS: Koch's postulates fulfilled.

text: Proof that a newly identified coronavirus, severe acute respiratory syndrome coronavirus (SARS-CoV) is the primary cause of severe acute respiratory syndrome (SARS) came from a series of studies on experimentally infected cynomolgus macaques (Macaca fascicularis). SARS-CoV-infected macaques developed a disease comparable to SARS in humans; the virus was re-isolated from these animals and they developed SARS-CoV-specific antibodies. This completed the fulfilment of Koch's postulates, as modified by Rivers for viral diseases, for SARS-CoV as the aetiological agent of SARS. Besides the macaque model, a ferret and a cat model for SARS-CoV were also developed. These animal models allow comparative pathogenesis studies for SARS-CoV infections and testing of different intervention strategies. The first of these studies has shown that pegylated interferon-alpha, a drug approved for human use, limits SARS-CoV replication and lung damage in experimentally infected macaques. Finally, we argue that, given the worldwide nature of the socio-economic changes that have predisposed for the emergence of SARS and avian influenza in Southeast Asia, such changes herald the beginning of a global trend for which we are ill prepared.


title: Middle East Respiratory Syndrome coronavirus (MERS-CoV) serology in major livestock species in an affected region in Jordan, June to September 2013.

text: Between June and September 2013, sera from 11 dromedary camels, 150 goats, 126 sheep and 91 cows were collected in Jordan, where the first human Middle-East respiratory syndrome (MERS) cluster appeared in 2012. All sera were tested for MERS-coronavirus (MERS-CoV) specific antibodies by protein microarray with confirmation by virus neutralisation. Neutralising antibodies were found in all camel sera while sera from goats and cattle tested negative. Although six sheep sera reacted with MERS-CoV antigen, neutralising antibodies were not detected.


title: Infectious bronchitis virus E protein is targeted to the Golgi complex and directs release of virus-like particles.

text: The coronavirus E protein is a poorly characterized small envelope protein present in low levels in virions. We are interested in the role of E in the intracellular targeting of infectious bronchitis virus (IBV) membrane proteins. We generated a cDNA clone of IBV E and antibodies to the E protein to study its cell biological properties in the absence of virus infection. We show that IBV E is an integral membrane protein when expressed in cells from cDNA. Epitope-specific antibodies revealed that the C terminus of IBV E is cytoplasmic and the N terminus is translocated. The short luminal N terminus of IBV E contains a consensus site for N-linked glycosylation, but the site is not used. When expressed using recombinant vaccinia virus, the IBV E protein is released from cells at low levels in sedimentable particles that have a density similar to that of coronavirus virions. The IBV M protein is incorporated into these particles when present. Indirect immunofluorescence microscopy showed that E is localized to the Golgi complex in cells transiently expressing IBV E. When coexpressed with IBV M, both from cDNA and in IBV infection, the two proteins are colocalized in Golgi membranes, near the coronavirus budding site. Thus, even though IBV E is present at low levels in virions, it is apparently expressed at high levels in infected cells near the site of virus assembly.


title: [Severe acute respiratory syndrome: a global overview of the epidemic].

text: In early February 2003, the World Health Organization (WHO) began receiving reports of patients with a syndrome characterized by an atypical pneumonia with rapid progression to respiratory failure without an identified cause despite extensive diagnostic workups. Most of these reports pointed out that the outbreak started in Southern China, specifically in the Guandong Province. The initial outbreak in South East Asia has already spread to other Regions in Asia, Europe, North and South America, and South Africa. Many of these cases can be linked through chains of transmission to an index case from the Guandong Province who visited Hong Kong. Although the exact mode of transmission has not been clearly established, the etiology of this syndrome has already been identified. A novel Coronavirus has been identified by electron microscopy and molecular assays in multiple laboratories from respiratory specimens throughout the world. The syndrome has been defined as SARS (Severe Acute Respiratory Syndrome) by WHO, and is characterized by an incubation period between 1 and 10 days (average 5 days) and by a febrile phase that usually lasts approximately 3 days. During the respiratory phase that begins around day 3, patients start developing a dry cough, shortness of breath and hypoxemia. Mechanical ventilatory support is required in about 10 to 40% of cases and the case-fatality rate ranges between 3 and 16%. The laboratory findings in SARS cases include leukopenia, thrombocytopenia, and a rise in transaminases and lactic dehydrogenase levels. Treatment of SARS includes supportive measures and the empiric use of ribavirin. Respiratory isolation, use of respiratory masks, and compulsory hand hygiene constitute the principal preventive measures. The confirmation of a case can be performed at reference laboratories by serologic and molecular assays. From the onset of this epidemic Mexico established a surveillance system as well as clinical guidelines and recommendations for the identification, prevention of secondary spread, and medical management of suspicious and probable cases by health care personnel.


title: Experimental demyelination produced by the A59 strain of mouse hepatitis virus.

text: Intracerebral inoculation of 4- to 6-week-old C57BL/6 mice with the A59 strain of mouse hepatitis virus (MHV), a murine coronavirus, produced biphasic disease. Acute hepatitis and mild meningoencephalitis were followed by subacute spastic paralysis with demyelinating lesions in the brain and spinal cord as determined by Epon-embedded toluidine-blue-stained sections and by electronmicroscopy. MHV-A59 was cultured by plaque assay from the blood, brain, spinal cord, and liver of infected mice during the acute phase, but not in the chronic stage. MHV-A59 antigen was detected by immunofluorescence (IF) until 3 months postinfection (PI). Serum anti-MHV-A59 antibodies were detected from 7 days to 5 months PI. The induction of demyelination by MHV-A59 provides a suitable system to study virus-induced demyelination further.


title: Gill-associated nidovirus of Penaeus monodon prawns transcribes 3'-coterminal subgenomic mRNAs that do not possess 5'-leader sequences.

text: Sequence analysis of the approximately 20 kb 5'-terminal portion of the ssRNA genome of gill-associated virus (GAV) of Penaeus monodon prawns has previously established that it contains an ORF1a-1b replicase gene equivalent to those of the coronavirus and arterivirus members of the order Nidovirales. Sequence analysis of the remaining approximately 6.2 kb of the GAV genome downstream of ORF1a-1b to a 3'-poly(A) tail has identified two highly conserved intergenic sequences in which 29/32 nucleotides are conserved. Northern hybridization using probes to the four putative GAV ORFs and either total or poly(A)-selected RNA identified two 3'-coterminal subgenomic (sg) mRNAs of approximately 6 kb and approximately 5.5 kb. Primer extension and 5'-RACE analyses showed that the sgmRNAs initiate at the same 5'-AC positions in the central region of the two conserved intergenic sequences. Neither method provided any evidence that the GAV sgmRNAs are fused to genomic 5'-leader RNA sequences as is the case with vertebrate coronaviruses and arteriviruses. Intracellular double-stranded (ds)RNAs equivalent in size to the 26.2 kb genomic RNA and two sgRNAs were also identified by RNase/DNase digestion of total RNA from GAV-infected prawn tissue. The identification of only two sgmRNAs that initiate at the same position in conserved intergenic sequences and the absence of 5'-genomic leader sequences fused to these sgmRNAs confirms that GAV has few genes and suggests that it utilizes a transcription mechanism possibly similar to the vertebrate toroviruses but distinct from coronaviruses and arteriviruses.


title: The role of wild mammals in the maintenance of Rift Valley fever virus.

text: Rift Valley fever virus (RVFV) is a zoonotic arbovirus affecting primarily domestic ruminants and humans. Numerous vector species are known or implicated in the transmission of RVFV. The role of mammals in the maintenance of RVFV, and the existence of a wild mammal reservoir in the epidemiologic cycle of RVFV, remain largely unknown. Our objective is to present a detailed review of studies undertaken on RVFV, often associated with wild mammals, with the aim of focusing future research on potential reservoirs of the virus. Natural and experimental infections related to RVFV in several mammalian orders, including Artiodactyla, Chiroptera, Rodentia, Primata (nonhuman), Perissodactyla, Carnivora, Proboscidea, Erinaceomorpha, and Lagomorpha, are reviewed; the first four orders have received the greatest attention. The possible role of wild ruminants, especially African buffalo (Syncerus caffer), is also discussed. Conflicting results have been published concerning rodents but, based on the literature, the likely candidate species include the African genera Arvicanthis and Micaelamys and the widely introduced roof rat (Rattus rattus). Members of the orders Chiroptera and Rodentia should receive greater attention associated with new research programs. For the other orders mentioned above, few data are available. We are unaware of any investigation concerning the orders Afrosoricida and Soricomorpha, which are represented in the geographic area of RVFV and can be abundant. As a first step to resolve the question of wild mammals as a reservoir of RVFV, serologic and virologic surveys should be promoted during epizootic periods to document infected wild animals and, in the case of positive results, extended to interepidemic periods to explore the role of wild animals as possible reservoirs.


Unconventional Agents and Unclassified Viruses: Recent Advances in Biology and Epidemiology. Proceedings of the 8th Munich Symposium on Microbiology. Munich, October 21-22, 1992.


title: Impact of attending a health care conference in Toronto during the severe acute respiratory syndrome crisis: survey of delegates.

text: OBJECTIVE To describe the impact on delegates of attending the Canadian Pain Society's annual meeting in Toronto during the severe acute respiratory syndrome (SARS) crisis in May 2003. METHODS A prospective survey design was used. Surveys were sent to all delegates (n=432) who attended the conference, and 294 delegates responded (68% response rate). The survey was developed to determine the level of concern about travelling to Toronto; the adequacy of screening measures; the level of stress about attending; and the occupational consequences of attending. RESULTS Fifty per cent of the participants were not concerned about travelling to Toronto, while the other 50% expressed some concern ranging from mild to serious. Concerns included being exposed to SARS and fear of transmitting it to others upon return. Reasons for attending the conference despite concern included a desire for continuing education, decrease in the number of reported SARS cases, and perceived minimal risk. Almost one-half (n=140) felt the screening measures at the conference were adequate, while 4% felt they were inadequate and 9% somewhat adequate. Delegates (n=99) suggested that temperature-taking (32.2%), improved screening vigilance (14.4%), SARS screening forms checked daily (9.1%), strictly controlled entry (8.1%) and adopting hospital screening procedures (7.1%) should have been instituted. CONCLUSION Health care professionals planning conferences in this era of new respiratory diseases can benefit from understanding the responses of delegates who attended conferences during outbreaks. Clear communication about the potential risks and benefits, as well as instituting full screening precautions, will help to allay concerns.


title: Cyclophilin A: A Key Factor in Virus Replication and Potential Target for Anti-viral Therapy.

text: Cyclophilin A (CypA) is a key member of immunophilins that has peptidyl-prolyl cis/trans isomerase (PPIase) activity. Besides acting as a cellular receptor for immunosuppressive drug cyclosporine A (CsA), CypA is involved in various cellular activities. CypA has an important role in viral infection which either facilitates or inhibits their replication. Inhibition of CypA via inhibitors is useful for overcoming several viral infections, indicating that CypA is an attractive target for anti-viral therapy. Collectively, these facts demonstrate the critical roles of CypA in mediating or inhibiting viral infections, suggesting that CypA can be an attractive cellular target for the development of anti-viral therapy.


title: Differential in vitro inhibition of feline enteric coronavirus and feline infectious peritonitis virus by actinomycin D.

text: The growth of feline enteric coronavirus strain 79-1683 in whole feline embryo cells was inhibited by the presence of 1 microgram/ml of actinomycin D in the culture fluid. No virus-specific mRNAs could be detected in such cultures and yields of infectious virus were depressed by > 99%. By contrast, the antigenically related feline infectious peritonitis virus strain 79-1146 was unaffected by the presence of actinomycin D, indicating a fundamental difference between the two feline coronavirus strains in their requirements for host-encoded function(s).


title: Influence of viral infections on body weight, survival, and tumor prevalence in Fischer 344/NCr rats on two-year studies.

text: Sendai virus (SV), pneumonia virus of mice (PVM), and rat coronavirus/sialodacryoadenitis virus (RCV/SDAV) were common viral infections of rats in the National Cancer Institute-National Toxicology Program (NCI-NTP) studies from 1977 to 1983. Influence of these viral infections on body weight, survival, and prevalences of spontaneous tumors in the F344/NCr rats of 28 diet control groups at five different laboratories were evaluated. Tumor prevalences evaluated in this investigation included the following: leukemia and tumors of the anterior pituitary, lungs, salivary glands and Harderian glands in both sexes; adrenal pheochromocytomas in male rats; and mammary tumors in female rats. SV and PVM but not RCV/SDAV infections were associated with significant (P less than 0.05) decreases in body weights of male and female rats. Male rat groups with PVM infection had a lower prevalence of leukemia and male rat groups with RCV/SDAV infection had a higher prevalence of anterior pituitary tumors than the corresponding uninfected groups. Female rat groups with SV infection had greater survival and a higher prevalence of lung tumors than groups without SV infection. However, none of the tumor prevalence and survival differences were statistically significant when interlaboratory variability and time-related effects were taken into account.


Australia antigen and hepatitis.


title: Assessment of febrile illness in the returned traveller.

text: BACKGROUND Fever is among the most frequently reported problems in returning travellers. OBJECTIVE This article provides an overview of the general approach to fever in the returned traveller, including identification of common causes and management. DISCUSSION The returned traveller may present with fever and it is important to exclude life threatening conditions such as malaria that may be related to the travel. A complete risk assessment should be undertaken, including a complete travel history, examination and further investigations, to help to narrow the differential diagnosis. Common tropical diseases found include malaria, dengue, enteric fever, rickettsial infections and respiratory infections. General practitioners should be alert to the public health implications of travel related diseases.


title: Microbial agents associated with waterborne diseases.

text: Many classes of pathogens excreted in feces are able to initiate waterborne infections. There are bacterial pathogens, including enteric and aquatic bacteria, enteric viruses, and enteric protozoa, which are strongly resistant in the water environment and to most disinfectants. The infection dose of viral and protozoan agents is lower than bacteria, in the range of one to ten infectious units or oocysts. Waterborne outbreaks of bacterial origin (particularly typhoid fever) in the developing countries have declined dramatically from 1900s. Therefore, some early bacterial agents such as Shigella sonnei remains prevalent and new pathogens of fecal origin such as zoonotic C. jejuni and E. coli O157:H7 may contaminate pristine waters through wildlife or domestic animal feces. The common feature of these bacteria is the low inoculum (a few hundred cells) that may trigger disease. The emergence in early 1992 of serotype O139 of V. cholerae with epidemic potential in Southeast Asia suggests that other serotypes than V. cholerae O1 could also getting on epidemic. Some new pathogens include environmental bacteria that are capable of surviving and proliferating in water distribution systems. Other than specific hosts at risk, the general population is refractory to infection with ingested P. aeruginosa. The significance of Aeromonas spp. in drinking water to the occurrence of acute gastroenteritis remains a debatable point and has to be evaluated in further epidemiological studies. Legionella and Mycobacterium avium complex (MAC) are environmental pathogens that have found an ecologic niche in drinking and hot water supplies. Numerous studies have reported Legionnaires' disease caused by L. pneumophila occurring in residential and hospital water supplies. M. avium complex frequently causes disseminated infections in AIDS patients and drinking water has been suggested as a source of infection; in some cases the relationship has been proven. More and more numerous reports show that Helicobacter pylori DNA can be amplified from feces samples of infected patients, which strongly suggests fecal-to-oral transmission. Therefore, it is possible that H. pylori infection is waterbome, but these assumptions need to be substantiated. Giardiasis has become the most common cause of human waterborne disease in the U.S. over the last 30 years. However, as a result of the massive outbreak of waterborne cryptosporidiosis in Milwaukee, Wisconsin, affecting an estimated 403,000 persons, there is increasing interest in the epidemiology and prevention of new infection disease caused by Cryptosporidium spp. as well as monitoring water quality. The transmission of Cryptosporidium and Giardia through treated water supplies that meet water quality standards demonstrates that water treatment technologies have become inadequate, and that a negative coliform no longer guarantees that water is free from all pathogens, especially from protozoan agents. Substantial concern persists that low levels of pathogen occurrence may be responsible for the endemic transmission of enteric disease. In addition to Giardia and Cryptosporidium, some species of genera Cyclospora, Isospora, and of family Microsporidia are emerging as opportunistic pathogens and may have waterborne routes of transmission. More than 15 different groups of viruses, encompassing more than 140 distinct types can be found in the human gut. Some cause illness unrelated with the gut epithelium, such as Hepatitis A virus (HAV) and Hepatitis E virus (HEV). Numerous large outbreaks have been documented in the U.S. between 1950 and 1970, and the incidence rate has strongly declined in developing countries since the 1970s. Hepatitis E is mostly confined to tropical and subtropical areas, but recent reports indicate that it can occur at a low level in Europe. A relatively small group of viruses have been incriminated as causes of acute gastroenteritis in humans and fewer have proven to be true etiologic agents, including rotavirus, calicivirus, astrovirus, and some enteric adenovirus. These enteric viruses have infrequently been identified as the etiologic agents of waterborne disease outbreaks, because of inadequate diagnostic technology, but many outbreaks of unknown etiology currently reported are likely due to viral agents. Actually, Norwalk virus and Norwalk-like viruses are recognized as the major causes of waterborne illnesses world-wide. The global burden of infectious waterborne disease is considerable. Reported numbers highly underestimate the real incidence of waterborne diseases. The most striking concern is that enteric viruses such as caliciviruses and some protozoan agents, such as Cryptosporidium, are the best candidates to reach the highest levels of endemic transmission, because they are ubiquitous in water intended for drinking, being highly resistant to relevant environmental factors, including chemical disinfecting procedures. Other concluding concerns are the enhanced risks for the classic group of debilitated subjects (very young, old, pregnant, and immunocompromised individuals) and the basic requirement of to take specific measures aimed at reducing the risk of waterborne infection diseases in this growing, weaker population.


title: Hajj pilgrims knowledge about Middle East respiratory syndrome coronavirus, August to September 2013.

text: In preparation for Hajj 2013, 360 French pilgrims were interviewed regarding their knowledge about Middle East respiratory syndrome (MERS). Respondents were aged 20–85 years, male-female ratio was 1.05:1;64.7% were aware of the MERS situation in Saudi Arabia; 35.3% knew about the Saudi Ministry of Health recommendations for at-risk pilgrims to postpone participation in the 2013 Hajj. None of 179 at-risk individuals(49.9%) decided to cancel their Hajj participation even after advice during consultation.


title: Requirement of proteolytic cleavage of the murine coronavirus MHV-2 spike protein for fusion activity.

text: The spike (S) protein of a non-fusogenic murine coronavirus, MHV-2, was compared to that of a variant, MHV-2f, with fusion activity. Two amino acids differed between The S proteins of these viruses; one was located in the signal sequence (amino acid 12) and the other in the putative cleavage site (amino acid 757). To determine which one of these amino acid changes is important for the alteration of fusogenicity, chimeric S proteins between MHV-2 and -2f were constructed and expressed in DBT cells by a vaccinia virus expression system. The results revealed that one amino acid change (Ser to Arg) at position 757 is responsible for the acquisition of fusogenicity of the MHV-2f S protein. This change also altered the susceptibility to proteolytic cleavage of the MHV-2 S protein which was originally uncleavable. We concluded that the non-fusogenic activity of MHV-2 results from the lack of cleavage of its S protein.


title: [Impact of COVID-19 during de-escalation in pediatric urology.]

text: Medical and surgical priorities have changed dramatically following the COVID-19 pandemic declaration. The rapid spread of the virus and the high number of cases has saturated the health system in our country and has forced many hospitals to redistribute resources to care for infected patients. This has led to asignificant reduction in surgical activity, in some cases reaching the point of delaying all elective interventions by performing only urgent interventions. The decrease in the number of infections with the progressive desaturation of hospitals has currently allowed us to enter a new phase of "de-escalation" or transition in order to recover our surgical activity in pediatric urology, which was practically canceled. It is proposed how surgical care activities such as outpatient care should be deal twith, in addition to the different circuits that patients must maintain and, above all, their and professional safety.


title: Duration of protection from reinfection following exposure to sialodacryoadenitis virus in Wistar rats.

text: Wistar rats [Cr1:(WI)BR] were inoculated intranasally with approximately 10(3) median mouse lethal infective doses of sialodacryoadenitis (SDA) virus. Animals were subsequently selected at random, removed to a separate isolation room, and reinfected with SDA virus at 3, 6, 9, 12 or 15 months. Pre- and postinoculation serum samples were collected from all animals during the course of the study and evaluated for antibody titers to SDA virus. All experimental, control and sentinel animals, following inoculation with SDA virus, were necropsied and examined for lesions consistent with SDA. Salivary gland lesions were minimal to absent in rats reinfected with SDA virus for up to 12 to 15 months after the initial exposure and minimal to moderate in the respiratory tract at 12 or 15 months. SDA-associated lesions were extensive in age matched control animals examined at each time period of reinfection with SDA virus. Thus, prior exposure to SDA virus did protect against the development of typical salivary gland lesions for up to 15 months. Recovered animals were evaluated for their ability to transmit the virus following reinfection. Rats reinfected at 6 or 9 months were infectious to their naive cage mates. The results indicate that reinfection with homologous rat coronavirus can occur as early as 6 months after the initial infection, and such rats can transmit the infection to contact controls.


title: Integrating occupational health with mainstream public health in Massachusetts: an approach to intervention.

text: In the late 19th century, workers' health was among the central concerns of the social reform movement to improve public health. Today, few state health agencies have comprehensive occupational health programs. Yet, state public health agencies have critical roles to play in occupational health and may be particularly instrumental in addressing the occupational health needs of underserved worker populations. Since the mid-1980s, with support from the National Institute for Occupational Safety and Health, the Massachusetts Department of Public Health has been working to build an occupational health program and promote the integration of occupational health concerns with ongoing public health activities in the state. This article provides a framework for considering the range of integration activities and presents examples of successful occupational health integration efforts in Massachusetts.


title: Brain natriuretic peptide and NT-proBNP levels reflect pulmonary artery systolic pressure in trekkers at high altitude.

text: Our objective was to evaluate the utility of the natriuretic peptides BNP (brain natriuretic peptide) and NT-proBNP as markers of pulmonary artery systolic pressure (PASP) in trekkers ascending to high altitude (HA). 20 participants had BNP and NT-proBNP assayed and simultaneous echocardiographic assessment of PASP performed during a trek to 5150 m. PASP increased significantly (p=0.006) with ascent from 24+/-4 to 39+/-11 mm Hg at 5150 m. At 5150 m those with a PASP>/=40 mm Hg (n=8) (versus those with PASP<40 mm Hg) had higher post-exercise BNP (pg/ml): 54.5+/-36 vs. 13.4+/-17 (p=0.012). Their resting BNP at 5150 m was also higher: 57.3+/-43.4 vs. 12.6+/-13 (p=0.017). In those with a pathological (>/=400 pg/ml) rise in NT-proBNP at 5150 m (n=4) PASP was significantly higher: 45.9+/-7.5 vs. 32.2+/-6.2 mm Hg (p=0.015). BNP and NT-proBNP may reflect elevated PASP, a central feature of high altitude pulmonary oedema, at HA.


title: Expression of bacteriophage T7 RNA polymerase in avian and mammalian cells by a recombinant fowlpox virus.

text: The bacteriophage T7 RNA polymerase gene was integrated into the fowlpox virus genome under the control of the vaccinia virus early/late promoter, P7.5. The recombinant fowlpox virus, fpEFLT7pol, stably expressed T7 RNA polymerase in avian and mammalian cells, allowing transient expression of transfected genes under the control of the T7 promoter. The recombinant fowlpox virus expressing T7 RNA polymerase offers an alternative to the widely used vaccinia virus vTF7-3, or the recently developed modified vaccinia virus Ankara (MVA) T7 RNA polymerase recombinant, a highly attenuated strain with restricted host-range. Recombinant fowlpox viruses have the advantage that as no infectious virus are produced from mammalian cells they do not have to be used under stringent microbiological safety conditions.


title: Efficacy of a paracetamol-pseudoephedrine combination for treatment of nasal congestion and pain-related symptoms in upper respiratory tract infection.

text: OBJECTIVE This study compared the efficacy of 1000 mg of paracetamol combined with 60 mg of pseudoephedrine, with that of either paracetamol or pseudoephedrine alone and placebo for the treatment of symptomatic URTI. RESEARCH DESIGN AND METHODS A double-blind, parallel group study was performed on 305 patients with URTI (nasal airflow resistance [NAR] of > 0.25 Pa cm3 s and a global pain score of at least moderate intensity). NAR and pain relief/intensity scores were measured over 4 h after initial dose. Patients then dosed up to three times daily for 3 days and recorded nasal congestion and pain intensity scores. MAIN OUTCOME MEASURES Nasal airflow conductance (NAC) and pain relief after the initial dose were primary objectives. NAC was calculated from NAR. Pain relief was measured on a 5-point verbal rating scale (VRS) and pain intensity and nasal congestion on a 4-point VRS. Data were analysed using analysis of covariance. Safety was assessed by adverse events. RESULTS A single dose of the combination was superior to paracetamol and placebo for NAC (p = 0.0001) and was superior to pseudoephedrine and placebo for pain relief (p < or = 0.048). Multiple doses of the combination were also superior to paracetamol and placebo for decongestion (p < or = 0.021) and were superior to pseudoephedrine and placebo for pain reduction (p < or = 0.0057). All treatments were well tolerated. CONCLUSIONS The combination treatment provided a greater decongestant effect than either paracetamol or placebo and better pain relief than either pseudoephedrine or placebo. The additive effect of the combination was apparent for both single and multiple doses.


title: Serosurvey of leptospirosis in feral hogs (Sus scrofa) in Florida.

text: Leptospira is a global pathogen of emerging public health importance in both developing and industrialized nations and can infect almost all mammalian species, including humans. As suburbanization and the popularity of outdoor recreational activities increases, so do human-wildlife and companion animal-wildlife interfaces. Florida offers a tropical climate favorable for outdoor activities and a semirural landscape that sustains an abundant feral hog population. Because no survey ofleptospirosis in feral hogs (Sus scrofa) in Florida has been published to our knowledge, we sought to establish preliminary seroprevalence ofleptospirosis exposure in feral hogs in Florida. Blood samples were collected opportunistically from 158 male and 166 female feral hogs taken at managed hunts and by permitted trappers in the northern, central, and southern regions of Florida. Samples were then analyzed using the microscopic agglutination test (MAT) for antibody titers to 20 Leptospira serovars representing 17 serogroups. A titer of > 1:100 was considered positive; 33% (107/324 total samples) were positive to at least one serovar, and 46% of those were positive to multiple serovars. Antibodies to L. interrogans serovar Bratislava strain Jez Bratislava (serogroup Australis) was the most common, with 18% (58/324) testing positive for antibodies. These initial data indicate that there is a significant possibility of feral hogs having a larger role in the complex etiology of leptospirosis in Florida than historically estimated and that further investigation is warranted.


title: Coronavirus JHM: cell-free synthesis of structural protein p60.

text: Sac(-) cells infected with murine coronavirus strain JHM shut off host cell protein synthesis and synthesized polypeptides with molecular weights of 150,000, 60,000, and 23,000. The 60,000- and 23,000-molecular-weight polypeptides comigrated with virion structural proteins p60 and p23, and the 60,000-molecular-weight protein was identified as p60 by tryptic peptide fingerprinting. Polyadenylate-containing RNA [poly(A) RNA] extracted from the cytoplasm of infected cells directed the synthesis of both 60,000- and 23,000-molecular-weight polypeptides in messenger-dependent cell-free systems derived from mouse L-cells and rabbit reticulocytes. The reticulocyte system also synthesized a 120,000-molecular-weight polypeptide that was specifically immunoprecipitated by antiserum raised against JHM virions. The identity of the 60,000- and 23,000-molecular-weight in vitro products was established by comigration with virion proteins, immunoprecipitation, and in the case of p60, tryptic peptide fingerprinting. The cytoplasmic poly(A) RNAs which encoded p60 and p23 sedimented in sucroseformamide gradients at 17S and 19S, respectively, and were clearly separable. These RNAs were among the major poly(A) RNA species synthesized in the cytoplasm of actinomycin D-treated cells late in infection, and the in vitro translation of size-fractionated RNA released from polysomes confirmed that they represent physiological mRNA's. These results suggest that the expression of the coronavirus JHM genome involves more than one subgenomic mRNA.


title: Reactive oxygen species induce virus-independent MAVS oligomerization in systemic lupus erythematosus.

text: The increased expression of genes induced by type I interferon (IFN) is characteristic of viral infections and systemic lupus erythematosus (SLE). We showed that mitochondrial antiviral signaling (MAVS) protein, which normally forms a complex with retinoic acid gene I (RIG-I)-like helicases during viral infection, was activated by oxidative stress independently of RIG-I helicases. We found that chemically generated oxidative stress stimulated the formation of MAVS oligomers, which led to mitochondrial hyperpolarization and decreased adenosine triphosphate production and spare respiratory capacity, responses that were not observed in similarly treated cells lacking MAVS. Peripheral blood lymphocytes of SLE patients also showed spontaneous MAVS oligomerization that correlated with the increased secretion of type I IFN and mitochondrial oxidative stress. Furthermore, inhibition of mitochondrial reactive oxygen species (ROS) by the mitochondria-targeted antioxidant MitoQ prevented MAVS oligomerization and type I IFN production. ROS-dependent MAVS oligomerization and type I IFN production were reduced in cells expressing the MAVS-C79F variant, which occurs in 30% of sub-Saharan Africans and is linked with reduced type I IFN secretion and milder disease in SLE patients. Patients expressing the MAVS-C79F variant also had reduced amounts of oligomerized MAVS in their plasma compared to healthy controls. Together, our findings suggest that oxidative stress-induced MAVS oligomerization in SLE patients may contribute to the type I IFN signature that is characteristic of this syndrome.


title: Non-invasive Measures of Core Temperature versus Ingestible Thermistor during Exercise in the Heat.

text: The accuracy of core temperature (Tc) thermometry from temporal, tympanic, and oral thermometry devices has been variable during exercise in a hot, humid environment. The purpose of the present study was to cross-validate temporal, two tympanic devices, and oral devices compared to an ingestible thermistor during exercise in a hot, humid environment. Fourteen young, active adults (6 women) completed a graded exercise test until voluntary exhaustion in an environmental chamber (35.5 ± 0.6 °C, 53.9 ± 5.8 % RH). There was no statistical difference in mean temperature between tympanic device 1 and pill-based core temperature (PBTc) measurements across all time points and were positively correlated (0.357; P<0.001). Temperatures of tympanic device 2 were statistically higher than PBTc (37.8 ± 0.7 ºC vs. 37.6 ± 1.0 ºC; respectfully) (P=0.008). At all time points, temperatures for the second tympanic device and PBTc were positively correlated (0.192; P=0.043). Temporal and PBTc values did not differ across time points and were positively correlated (0.262; P=0.005) across all time points. Mean oral temperature was significantly less than mean PBTc across all time points. (37.0 ± 0.4 ºC vs. 37.6 ± 1.0 ºC, respectively) (P<0.001). Across all time points, oral and PBTc were positively correlated (0.262; P=0.010). Tympanic and temporal devices can reflect Tc while exercising in a hot, humid environment. However, care should be taken when selecting the tympanic or temporal measurement device and validation is advised prior to heat illness mitigation in the field.


title: Oncologic specimen from laparoscopic assisted gastrectomy for gastric adenocarcinoma is comparable to D1-open surgery: the experience of a Canadian centre.

text: BACKGROUND The Eastern experience has reported the safety of laparoscopic assisted gastrectomy (LAG) for gastric cancer. Its use in Western countries is still debated owing to concerns about its oncologic equivalence to open gastrectomy (OG). We sought to review and compare their operative outcomes and oncologic specimen quality (number of harvested lymph nodes and surgical margins) for gastric adenocarcinoma (GA). METHODS We reviewed the charts of all patients undergoing LAG (2007-2010) and OG (2000-2010) for GA in a single institution. Several surgeons performed the OGs, whereas 1 fellowship-trained laparoscopic surgeon performed LAGs. The primary outcome was quality of the surgical specimen, assessed by the number of harvested lymph nodes (LNs) and margin status. Secondary outcomes were perioperative events. Data were analyzed as intention to treat. RESULTS We retrieved 60 cases (47 OGs, 13 LAGs). The conversion rate was 23%. Mean operative time was 115 minutes longer and blood loss was 425 mL less (both p < 0.001) for LAGs. A mean of 14.4 (standard deviation [SD] 9.8) and 11.2 (SD 8.2) LNs were harvested for OGs and LAGs, respectively (p = 0.29). Negative margins were achieved for all patients. Mean length of stay was similar (LAG: 19 d v. OG: 18.9 d; p = 0.91). The groups did not differ on major postoperative complications (12.7% v. 23.1%; p = 0.39) or operative mortality (2.1% v. 7.7%; p = 0.32). CONCLUSION Laparoscopic assisted gastrectomy is a challenging but safe and feasible procedure in experienced hands. It offers the same radical resection as OG regarding negative margins and LN retrieval. Long-term follow-up is warranted.


title: Estimating wildlife disease dynamics in complex systems using an Approximate Bayesian Computation framework.

text: Emerging infectious diseases of wildlife are of increasing concern to managers and conservation policy makers, but are often difficult to study and predict due to the complexity of host-disease systems and a paucity of empirical data. We demonstrate the use of an Approximate Bayesian Computation statistical framework to reconstruct the disease dynamics of bovine tuberculosis in Kruger National Park's lion population, despite limited empirical data on the disease's effects in lions. The modeling results suggest that, while a large proportion of the lion population will become infected with bovine tuberculosis, lions are a spillover host and long disease latency is common. In the absence of future aggravating factors, bovine tuberculosis is projected to cause a lion population decline of ~3% over the next 50 years, with the population stabilizing at this new equilibrium. The Approximate Bayesian Computation framework is a new tool for wildlife managers. It allows emerging infectious diseases to be modeled in complex systems by incorporating disparate knowledge about host demographics, behavior, and heterogeneous disease transmission, while allowing inference of unknown system parameters.


title: Comparative study of minimally invasive versus open esophagectomy for esophageal cancer in a single cancer center.

text: BACKGROUND In order to minimize the injury reaction during the surgery and reduce the morbidity rate, hence reducing the mortality rate of esophagectomy, minimally invasive esophagectomy (MIE) was introduced. The aim of this study was to compare the postoperative outcomes in patients with esophageal squamous cell carcinoma undergoing minimally invasive or open esophagectomy (OE). METHODS The medical records of 176 consecutive patients, who underwent minimally invasive esophagectomy (MIE) between January 2009 and August 2013 in Cancer Institute & Hospital, Chinese Academy of Medical Sciences, were retrospectively reviewed. In the same period, 142 patients who underwent OE, either Ivor Lewis or McKeown approach, were selected randomly as controls. The clinical variables of paired groups were compared, including age, sex, Charlson score, tumor location, duration of surgery, number of harvested lymph nodes, morbidity rate, the rate of leak, pulmonary morbidity rate, mortality rate, and hospital length of stay (LOS). RESULTS The number of harvested lymph nodes was not significantly different between MIE group and OE group (median 20 vs. 16, P = 0.740). However, patients who underwent MIE had longer operation time than the OE group (375 vs. 300 minutes, P < 0.001). Overall morbidity, pulmonary morbidity, the rate of leak, in-hospital death, and hospital LOS were not significantly different between MIE and OE groups. Morbidities including anastomotic leak and pulmonary morbidity, inhospital death, hospital LOS, and hospital expenses were not significantly different between MIE and OE groups as well. CONCLUSIONS MIE and OE appear equivalent with regard to early oncological outcomes. There is a trend that hospital LOS and hospital expenses are reduced in the MIE group than the OE group.


title: Immune response to orally administered calf reovirus-like agent and coronavirus vaccine.

text: Twenty 6-to 7-hour-old gnotobiotic calves inoculated orally with attenuated calf diarrhea reovirus-like agent and challenge-inoculated with virulent virus 48-72 h post-inoculation (PI) remained clinically normal during the post-vaccination observation period; one developed mild diarrhea after challenge inoculation. Four non-vaccinated challenge control calves developed severe diarrhea. Twenty 6-to 7-hour-old gnotobiotic calves inoculated orally with attenuated calf diarrhea coronavirus and challenge inoculated with virulent virus 96 h later remained clinically normal during the post-vaccination and post-challenge observation period. Four non-vaccinated challenge control calves developed severe diarrhea and 2 of these died. Five-foot long isolated loops prepared in the lower ileum (Thiry-Vella loop) of newborn colostrum-deprived calves were inoculated with attenuated coronavirus. Daily loop washings were cultured for virus and tested for neutralizing antibody. Peak viral titers of 10(6.5) to 10(7) occurred 3-4 days PI and descended rapidly to 0 between 6 and 8 days PI. Neutralizing antibody was first detected in the washings 6-8 days PI and reached a titer of 128-256 7 to 9 days PI. Loop immunoglobulin separated by gel filtration and identified by immunodiffusion were primarily IgM and IgA. Initial resistance to virulent viral infection is thought to be due to an interference phenomenon, with later resistance due to local antibody.


title: Real-time magnetic resonance imaging guidance improves the diagnostic yield of endomyocardial biopsy.

text: BACKGROUND Diagnostic yield of endomyocardial biopsy is low, particularly in disease that affects the myocardium in a non-uniform distribution. We hypothesized that real-time MRI guidance could improve the yield through targeted biopsy of focal myocardial pathology. METHODS An animal model of focal myocardial pathology was created by infusing 3mL of fluorescent microspheres (NuFlow Hydrocoat, 15μm diameter, 5 million spheres/mL) followed by 2mL of 100% ethanol to a branch coronary artery. Animals were survived for minimum 14days, before undergoing MRI guided endomyocardial biopsy using a custom 6.5Fr active visualization MRI-conditional bioptome and X-ray guided biopsy using a commercial bioptome. Specimens were analyzed using a dissecting microscope under ultraviolet light to determine the proportion of 'on-target' specimens containing fluorescent microspheres. RESULTS A total of 77 specimens were obtained using real-time MRI guidance and 87 using X-ray guidance, in five animals. Specimens obtained with the MRI-conditional bioptome were smaller compared with the commercial X-ray bioptome. Real-time MRI guidance significantly increased the diagnostic yield of endomyocardial biopsy (82% vs. 56% on-target biopsy specimens with real-time MRI vs. X-ray guidance, p<0.01). CONCLUSIONS Endomyocardial biopsy performed using real-time MRI guidance is feasible and significantly improves the diagnostic yield compared with X-ray fluoroscopy guidance.


Guideline for the management of acute diarrhea in adults.


title: Moving towards the global control of foot and mouth disease: an opportunity for donors.

text: Livestock contributes significantly to the world economy. However, animal diseases are still a major constraint on economic growth, the reduction of poverty and food security. Among the most significant diseases is foot and mouth disease (FMD), a highly contagious, multi-species animal disease with a devastating impact on national economies and trade. Less obvious is the severe constraint that FMD places on both development and the reduction of poverty in developing countries where this disease is endemic. As a result of its global implications and the high costs that it imposes on society, FMD is an infectious disease whose control and prevention are recognised as being a global public good. Moving towards the global control of FMD should be considered a priority for donors, but will require long-term commitment from all parties, strong political will from governments and concerted financial support from donors. Areas of intervention must fall within the framework of programmes developed by international organisations, such as the Food and Agriculture Organization of the United Nations (FAO) and the World Organisation for Animal Health (OIE), through the FAO/OIE Global Framework for the Progressive Control of FMD and Other Transboundary Animal Diseases, as well as the disease control programmes of the regions concerned. Such a goal should specifically focus on analytical work (micro-economic impact and cost-benefit analyses of FMD at the household level and on the poor), research, surveillance networks, communication, monitoring and evaluation, and continuous strengthening of Veterinary Services.


title: Trends in HIV- and STD-related risk behaviors among high school students--United States, 1991-2007.

text: Persons who engage in unprotected sexual intercourse or use injection drugs are at increased risk for human immunodeficiency virus (HIV) infection and sexually transmitted diseases (STDs). Changes in HIV- and STD-related risk behaviors among high school students in the United States during 1991-2005 were reported previously. To update these analyses through 2007, CDC analyzed data from nine biennial national Youth Risk Behavior Surveys (YRBS). This report summarizes the results of that analysis, which indicated that, during 1991-2007, the percentage of U.S. high school students who ever had sexual intercourse decreased 12%, the percentage who had sexual intercourse with four or more persons during their lifetime decreased 20%, and the percentage who were currently sexually active decreased 7%. Among students who were currently sexually active, the prevalence of condom use increased 33%. However, these changes in risk behaviors were not observed in some subgroups. In addition, no changes were detected in the prevalence of sexual risk behaviors from 2005 to 2007, and many students still engaged in behaviors that place them at risk for HIV infection and STDs. Additional efforts to reduce sexual risk behaviors, particularly among black, Hispanic, and male students, must be implemented to meet the Healthy People 2010 national health objective for adolescent sexual behaviors (objective no. 25-11) and to decrease rates of HIV infection and STDs.


title: Neither the RNA nor the proteins of open reading frames 3a and 3b of the coronavirus infectious bronchitis virus are essential for replication.

text: Gene 3 of infectious bronchitis virus is tricistronic; open reading frames (ORFs) 3a and 3b encode two small nonstructural (ns) proteins, 3a and 3b, of unknown function, and a third, structural protein E, is encoded by ORF 3c. To determine if either the 3a or the 3b protein is required for replication, we first modified their translation initiation codons to prevent translation of the 3a and 3b proteins from recombinant infectious bronchitis viruses (rIBVs). Replication in primary chick kidney (CK) cells and in chicken embryos was not affected. In chicken tracheal organ cultures (TOCs), the recombinant rIBVs reached titers similar to those of the wild-type virus, but in the case of viruses lacking the 3a protein, the titer declined reproducibly earlier. Translation of the IBV E protein is believed to be initiated by internal entry of ribosomes at a structure formed by the sequences corresponding to ORFs 3a and 3b. To assess the necessity of this mechanism, we deleted most of the sequence representing 3a and 3b to produce a gene in which ORF 3c (E) was adjacent to the gene 3 transcription-associated sequence. Western blot analysis revealed that the recombinant IBV produced fivefold less E protein. Nevertheless, titers produced in CK cells, embryos, and TOCs were similar to those of the wild-type virus, although they declined earlier in TOCs, probably due to the absence of the 3a protein. Thus, neither the tricistronic arrangement of gene 3, the internal initiation of translation of E protein, nor the 3a and 3b proteins are essential for replication per se, suggesting that these proteins are accessory proteins that may have roles in vivo.


title: Equine arteritis virus non-structural protein 1, an essential factor for viral subgenomic mRNA synthesis, interacts with the cellular transcription co-factor p100.

text: Non-structural protein 1 (nsp1), the N-terminal subunit of the replicase polyprotein of the arterivirus Equine arteritis virus (EAV), is essential for viral subgenomic mRNA synthesis, but fully dispensable for genome replication. However, at the molecular level, the role of nsp1 in EAV subgenomic mRNA synthesis is poorly understood. A yeast two-hybrid screen did not reveal interactions between EAV nsp1 and other viral non-structural proteins or the nucleocapsid protein, although both nsp1 and the nucleocapsid protein were found to form homomers. Subsequently, a yeast two-hybrid screen of a HeLa cell cDNA library was performed using nsp1 as bait. Remarkably, this analysis revealed (potential) interactions between EAV nsp1 and factors that are involved in host cell transcriptional regulation. The interaction of nsp1 with one of these proteins, p100, a transcription co-activator that also interacts with regulatory proteins of other viruses, was confirmed by mutual co-immunoprecipitation from lysates of EAV-susceptible mammalian cells.


title: Nosocomial transmission of Mycobacterium tuberculosis found through screening for severe acute respiratory syndrome--Taipei, Taiwan, 2003.

text: The emergence of severe acute respiratory syndrome (SARS) has highlighted the importance of hospital infection-control programs. Prevention of Mycobacterium tuberculosis transmission also requires effective infection control in health-care facilities. In Taipei, Taiwan, an area with moderate to high incidence of tuberculosis (TB) (50-74 cases per 100,000 population), health-care workers (HCWs) are at increased risk for M. tuberculosis (Taiwan Center for Disease Control, unpublished data, 2002). In April 2003, SARS-related screening in a hospital in Taipei resulted in the detection of suspected TB among HCWs. This report summarizes how SARS screening led to the discovery of 60 cases of TB. HCWs in Taiwan should remain vigilant for cases of TB so persons suspected of having TB are evaluated and treated promptly.


title: Comparative analysis of coronavirus JHM-induced demyelinating encephalomyelitis in Lewis and Brown Norway rats.

text: Lewis and Brown Norway rats were infected at different ages with the neurotropic murine coronavirus strain, JHM and the resultant central nervous system diseases were studied. Suckling rats of both strains came down with a fatal, acute encephalomyelitis. Weanling Lewis rats developed a subacute demyelinating encephalomyelitis which neuropathologically revealed changes of an immunopathologic reaction. In contrast, Brown Norway rats developed a clinically silent subacute demyelinating encephalomyelitis with a persistent JHM virus infection which was less severe and quite different from the subacute demyelinating encephalomyelitis in Lewis rats with respect to size, distribution, and localization of the demyelinating plaques as well as the type of infiltrating cells. In addition, infected Lewis rats showed a pronounced lymphocyte proliferation to myelin basic protein and JHM virus whereas lymphocytes from infected Brown Norway rats did not react to these two antigens. These observations demonstrate the pathogenetic importance of host factors in the development of virus-induced demyelination.


title: On-site quantification of re-entry ratio of ventilation exhausts in multi-family residential buildings and implications.

text: UNLABELLED In the worldwide spread of Severe Acute Respiratory Syndrome (SARS) in 2003, cluster of cases occurred in a number of large high-rise residential building blocks, especially in Hong Kong. In this study, we examined one of the most likely virus-spread mechanisms, which is related to the inter-flat or inter-zonal airflow through open-windows caused by buoyancy effects. Dual tracer gases of CO2 and SF6 are employed simultaneously to quantify the amount of the exhaust air coming out of the upper part of the window of a floor that re-enters the lower part of the open-window at the immediate upper floor. It was found that the room air could contain up to 7% of the exhaust air from the lower floor, and this occurs at low wind conditions with a combination of indoor-outdoor temperature difference. The results can well explain the earlier governmental finding that DNA strings of SARS Corono-Virus were detected within the sampled deposits on the window sills of the upper floors of the two index patients' flats. The preliminary and yet alarming conclusion may be that, in high-rise buildings, windows flush with a flat façade can be a major route for the vertical spread of pathogen-containing aerosols. PRACTICAL IMPLICATIONS The confirmation of the transmission route can have a number of implications to both building design and infectious disease control. With respect to the former, considerations should be given to minimize the inter-flat airflow via windows flush with façade for high-rise buildings with natural ventilation; and with respect to the latter, more targeted and earlier intervention can be implemented in case of any highly infectious disease outbreaks.


title: [Study on the dynamics of IgG antibody in 311 patients with severe acute respiratory syndrome].

text: OBJECTIVE To detect the level and dynamic change of severe acute respiratory syndrome (SARS)-coronavirus-specific IgG antibody in conavalescent SARS patients, and to provide information for prevention and vaccine development. METHODS IgG antibody against coronavirus was detected by ELISA in the blood of 311 convalescent SARS patients for every 2 - 4 weeks. Stata 7.0 statistics software was used to analyse the results. RESULTS IgG antibody was detected positive on each testing of all the convalescent patients and its peak appeared 35 days after recovery. IgG antibody level showed a 35.8% decrease within one year. CONCLUSION Data showed that all the SARS convalescent patients had generated high level of specific IgG antibody against coronavirus in the early stage of recovery, but the antibody level declined along with the progress of convalescence, suggesting that the detection of the IgG antibody should go on until it disappeared.


title: Alexia without agraphia associated with spleniogeniculate infarction.

text: Pure alexia, following an infarction in the distribution of the left posterior cerebral artery, is attributed to damage of the left occipital lobe and the splenium of the corpus callosum. We describe a case of pure alexia in a 57-year-old woman with infarction of the left lateral geniculate body and the splenium of the corpus callosum, a variation on this classic disconnection syndrome.


title: Regulation of IRF-3-dependent innate immunity by the papain-like protease domain of the severe acute respiratory syndrome coronavirus.

text: Severe acute respiratory syndrome coronavirus (SARS-CoV) is a novel coronavirus that causes a highly contagious respiratory disease, SARS, with significant mortality. Although factors contributing to the highly pathogenic nature of SARS-CoV remain poorly understood, it has been reported that SARS-CoV infection does not induce type I interferons (IFNs) in cell culture. However, it is uncertain whether SARS-CoV evades host detection or has evolved mechanisms to counteract innate host defenses. We show here that infection of SARS-CoV triggers a weak IFN response in cultured human lung/bronchial epithelial cells without inducing the phosphorylation of IFN-regulatory factor 3 (IRF-3), a latent cellular transcription factor that is pivotal for type I IFN synthesis. Furthermore, SARS-CoV infection blocked the induction of IFN antiviral activity and the up-regulation of protein expression of a subset of IFN-stimulated genes triggered by double-stranded RNA or an unrelated paramyxovirus. In searching for a SARS-CoV protein capable of counteracting innate immunity, we identified the papain-like protease (PLpro) domain as a potent IFN antagonist. The inhibition of the IFN response does not require the protease activity of PLpro. Rather, PLpro interacts with IRF-3 and inhibits the phosphorylation and nuclear translocation of IRF-3, thereby disrupting the activation of type I IFN responses through either Toll-like receptor 3 or retinoic acid-inducible gene I/melanoma differentiation-associated gene 5 pathways. Our data suggest that regulation of IRF-3-dependent innate antiviral defenses by PLpro may contribute to the establishment of SARS-CoV infection.


title: Caspase cleavage of the nonstructural protein NS1 mediates replication of Aleutian mink disease parvovirus.

text: Virus-induced apoptosis of infected cells can limit both the time and the cellular machinery available for virus replication. Hence, many viruses have evolved strategies to specifically inhibit apoptosis. However, Aleutian mink disease parvovirus (ADV) is the first example of a DNA virus that not only induces apoptosis but also utilizes caspase activity to facilitate virus replication. To determine the function of caspase activity during ADV replication, virus-infected cell lysates or purified ADV proteins were incubated with various purified caspases. Caspases cleaved the major nonstructural protein of ADV (NS1) at two caspase recognition sequences, whereas ADV structural proteins could not be cleaved. Importantly, the NS1 products could be identified in ADV-infected cells but were not present in infected cells pretreated with caspase inhibitors. By mutating putative caspase cleavage sites (D to E), we mapped the two cleavage sites to amino acid residues NS1:227 (INTD downward arrow S) and NS1:285 (DQTD downward arrow S). Replication of ADV containing either of these mutations was reduced 10(3)- to 10(4)-fold compared to that of wild-type virus, and a construct containing both mutations was replication defective. Immunofluorescent studies revealed that cleavage was required for nuclear localization of NS1. The requirement for caspase activity during permissive replication suggests that limitation of caspase activation and apoptosis in vivo may be a novel approach to restricting virus replication.


title: Passive protection of dogs against clinical disease due to Canine parvovirus-2 by specific antibody from chicken egg yolk.

text: The protective effect of immunoglobulins derived from chicken egg yolk (IgY) against infection by Canine parvovirus 2 (CPV-2) was evaluated in 10 beagle dogs orally challenged with a strain of the virus. The 2-mo-old dogs were divided into 3 groups and treated with powders containing CPV-2 IgY or normal egg yolk for 7 d after the challenge. The 4 dogs receiving normal egg yolk (control group) demonstrated mild symptoms typical of CPV-2 infection, such as vomiting, diarrhea, and weight loss. No symptoms were observed by 16 d after challenge in the 3 dogs receiving 2 g of IgY powder. Of the 3 dogs receiving 0.5 g of IgY powder, 2 had clinical CPV-2 disease; however, the manifestations were less severe than in the control group. Furthermore, the IgY-treated groups had significantly greater weight gain and shorter duration of virus shedding than the control group. These results indicate that IgY is useful in protecting dogs from CPV-2-induced clinical disease.


Intraductal papillary mucinous neoplasm.


Genetic variability of human respiratory coronavirus OC43.