title: Coronavirus-like virions associated with a wasting syndrome in guinea pigs.

text: An apparent wasting syndrome was observed in newly arriving 3 to 4 week old guinea pigs characterized by anorexia, weight loss, diarrhea, perineal staining and death. Diagnostic efforts to attribute the disease to husbandry, environmental factors or to known guinea pig pathogens were unsuccessful. Clinical signs, enteric histopathological lesions and diagnostic transmission electron microscopy identification of typical coronavirus-like virions in fecal samples were consistent with enteric coronaviral diseases seen in other species.


title: Boots for Achilles: progesterone's reduction of cholesterol is a second-order adaptation.

text: Progesterone and cholesterol are both vital to pregnancy. Among other functions, progesterone downregulates inflammatory responses, allowing for maternal immune tolerance of the fetal allograft. Cholesterol a key component of cell membranes, is important in intracellular transport, cell signaling, nerve conduction, and metabolism Despite the importance of each substance in pregnancy, one exercises an antagonistic effect on the other, as periods of peak progesterone correspond with reductions in cholesterol availability, a consequence of progesterone's negative effects on cholesterol biosynthesis. This arrangement is understandable in light of the threat posed by pathogens early in pregnancy. Progesterone-induced immunomodulation entails increased vulnerability to infection, an acute problem in the first trimester, when fetal development is highly susceptible to insult. Many pathogens rely on cholesterol for cell entry, egress, and replication. Progesterone's antagonistic effects on cholesterol thus partially compensate for the costs entailed by progesterone-induced immunomodulation. Among pathogens to which the host's vulnerability is increased by progesterone's effects, approximately 90% utilize cholesterol, and this is notably true of pathogens that pose a risk during pregnancy. In addition to having a number of possible clinical applications, our approach highlights the potential importance of second-order adaptations, themselves a consequence of the lack of teleology in evolutionary processes.


title: Inflammatory bowel diseases: multiple benefits from therapy with dipeptidyl- and alanyl-aminopeptidase inhibitors.

text: Inflammatory bowel diseases (IBD) are driven by imbalances in innate and acquired immune response. In IBD two dysregulated T cell subsets are in the focus of interest: activated effector T cells and regulatory T cells. These T cell subsets are characterized by a strong expression of the ectopeptidases dipeptidyl peptidase IV (DPIV /CD26) and aminopeptidase N (APN/CD13), which are thought to a role in the control of immune activation and in regulating cellular communication by hydrolyzing bioactive polypeptides. Since inhibitors of both enzymes were shown to be effective in limiting immune activation processes in vitro as well as in vivo, they emerged as new drug candidates for the treatment of diseases associated with an imbalanced T cell response, such as IBD. In this review we intent to throw light on the putative role of DPIV, APN and related enzymes in the regulation of immune and non-immune processes in inflammatory bowel diseases, on possible benefits from peptidase inhibitor therapy in these diseases as well on the gaps of knowledge in this field.


title: A historical sketch of the discovery and development of HIV-1 integrase inhibitors.

text: The long process of HIV-1 integrase inhibitor discovery and development can be attributed to both the complexity of HIV-1 integration and poor 'integration' of these researches into mainstream investigations on antiretroviral therapy in the mid-1990s. Of note, some fungal extracts investigated during this period contain the beta-hydroxyketo group, later recognised to be a key structural requirement for keto-enol acids (also referred to as diketo acids) and other integrase inhibitors. This review reconstructs (in the general context of the history of AIDS research) the principal steps that led to the integrase inhibitors currently in clinical trials, and discusses possible future directions.


Hyper-inflammatory Syndrome in a Child With COVID-19 Treated Successfully With Intravenous Immunoglobulin and Tocilizumab.


title: Mutational analysis of the virus and monoclonal antibody binding sites in MHVR, the cellular receptor of the murine coronavirus mouse hepatitis virus strain A59.

text: The primary cellular receptor for mouse hepatitis virus (MHV), a murine coronavirus, is MHVR (also referred to as Bgp1a or C-CAM), a transmembrane glycoprotein with four immunoglobulin-like domains in the murine biliary glycoprotein (Bgp) subfamily of the carcinoembryonic antigen (CEA) family. Other murine glycoproteins in the Bgp subfamily, including Bgp1b and Bgp2, also can serve as MHV receptors when transfected into MHV-resistant cells. Previous studies have shown that the 108-amino-acid N-terminal domain of MHVR is essential for virus receptor activity and is the binding site for monoclonal antibody (MAb) CC1, an antireceptor MAb that blocks MHV infection in vivo and in vitro. To further elucidate the regions of MHVR required for virus receptor activity and MAb CC1 binding, we constructed chimeras between MHVR and other members of the CEA family and tested them for MHV strain A59 (MHV-A59) receptor activity and MAb CC1 binding activity. In addition, we used site-directed mutagenesis to introduce selected amino acid changes into the N-terminal domains of MHVR and these chimeras and tested the abilities of these mutant glycoproteins to bind MAb CC1 and to function as MHV receptors. Several recombinant glycoproteins exhibited virus receptor activity but did not bind MAb CC1, indicating that the virus and MAb binding sites on the N-terminal domain of MHVR are not identical. Analysis of the recombinant glycoproteins showed that a short region of MHVR, between amino acids 34 and 52, is critical for MHV-A59 receptor activity. Additional regions of the N-terminal variable domain and the constant domains, however, greatly affected receptor activity. Thus, the molecular context in which the amino acids critical for MHV-A59 receptor activity are found profoundly influences the virus receptor activity of the glycoprotein.


title: The evolving role of nuclear molecular imaging in cancer.

text: BACKGROUND: Novel therapies targeted to specific tumor pathways are entering the clinic. The need for in vivo monitoring of resulting molecular changes, particularly with respect to the tumor microenvironment, is growing. Molecular imaging is evolving to include a variety of imaging methods to enable in vivo monitoring of cellular and molecular processes. OBJECTIVES: This article reviews the emerging role of molecular imaging in the development of improved therapeutic strategies that provide better patient selection for therapeutic personalization (i.e. determine which therapies have the greatest chance of success given the individual patient's disease genetic, and phenotypical profile). METHODS: In order to illustrate the utility of integrating molecular imaging into therapy development strategies, current and emerging applications of nuclear molecular imaging strategies will be compared with conventional strategies. Proposed methods of integrating molecular imaging techniques into cancer therapeutic development and limitations of these techniques will be discussed. RESULTS/CONCLUSION: Molecular imaging provides a variety of new tools to accelerate the development of cancer therapies. The recent drive to develop molecular imaging probes and standardize molecular imaging techniques is creating the scaffolding for the evolving paradigm shift to personalized cancer therapy.


title: Residues in the apical domain of the feline and canine transferrin receptors control host-specific binding and cell infection of canine and feline parvoviruses.

text: Canine parvovirus (CPV) and feline panleukopenia virus (FPV) capsids bind to the transferrin receptors (TfRs) of their hosts and use these receptors to infect cells. The binding is partially host specific, as FPV binds only to the feline TfR, while CPV binds to both the canine and feline TfRs. The host-specific binding is controlled by a combination of residues within a raised region of the capsid. To define the TfR structures that interact with the virus, we altered the apical domain of the feline or canine TfR or prepared chimeras of these receptors and tested the altered receptors for binding to FPV or CPV capsids. Most changes in the apical domain of the feline TfR did not affect binding, but replacing Leu221 with Ser or Asp prevented receptor binding to either FPV or CPV capsids, while replacing Leu221 with Lys resulted in a receptor that bound only to CPV but not to FPV. Analysis of recombinants of the feline and canine TfRs showed that sequences controlling CPV-specific binding were within the apical domain and that more than one difference between these receptors determined the CPV-specific binding of the canine TfR. Single changes within the canine TfR which removed a single amino acid insertion or which eliminated a glycosylation site gave that receptor the expanded ability to bind to FPV and CPV. In some cases, binding of capsids to mutant receptors did not result in infection, suggesting a structural role for the receptor in cell infection by the viruses.


title: Risk Factors for Severe Adenovirus Infection in Children during an Outbreak in Singapore.

text: BACKGROUND Human adenoviruses (HAdVs) can cause a variety of human illnesses, with associated temporal and geographic changes in disease incidence. We report the emergence of an outbreak of HAdV infections in Singapore, presumably caused by a change of the predominating type to HAdV-7. We examined the clinical features of children admitted with HAdV infection to 1 institution and the risk factors for severe infection. MATERIALS AND METHODS This is a retrospective case-control study of all HAdV-infected children admitted during weeks 1 to 19 in 2013, as identified from laboratory records. A descriptive retrospective analysis of epidemiology, clinical data and the outcome of these children was also performed. Patients with severe infections were defined as cases, those with non-severe infections as controls, and the 2 groups were compared to find possible independent risk factors. RESULTS Eighty-five patients with HAdV infection were studied, including 11 (12.9%) cases and 74 (87.1%) controls. Binary logistic regression showed that cases were more likely to be <2 years old (adjusted OR 10.6, 95% CI, 1.8 to 63.2) and to have significant comorbidities (adjusted OR 19.9, 95% CI, 3.4 to 116.1) compared to controls. The predominant type in 2013 was HAdV-7, which differed from 2011 and 2012, when HAdV-3 was more common. There was a trend towards pneumonia being more common in patients infected with HAdV-7 than in patients infected with other types, although this did not reach statistical significance (OR 2.8, 95% CI, 0.9 to 8.7). CONCLUSION The emergence of HAdV-7 in a population where other HAdV types had circulated previously may have caused the outbreak in Singapore, and this was associated with more serious infections in children. Young age (<2 years) and significant comorbidities were associated with more severe HAdV infection.


Infectious diseases. SARS experts want labs to improve safety practices.


title: Protein interactions during coronavirus assembly.

text: Coronaviruses assemble and obtain their envelope at membranes of the intermediate compartment between the endoplasmic reticulum and Golgi complex. Like other enveloped viruses, coronavirus assembly is presumably dependent on protein localization and protein-protein as well as protein-RNA interactions. We have used the bovine coronavirus (BCV) as a model to study interactions between the viral proteins in virus-infected cells that are important for coronavirus assembly. BCV is a prototype for the coronaviruses that express an additional major structural protein, the hemagglutinin esterase (HE), in addition to the spike (S) glycoprotein, membrane (M) glycoprotein, and nucleocapsid (N) protein. Complexes consisting of the M, S, and HE proteins were detected in virus-infected cells by coimmunoprecipitations. Kinetic analyses demonstrated that S protein and HE each quickly formed a complex with M protein after synthesis, whereas heterocomplexes consisting of all three proteins formed more slowly. The kinetics of HE biosynthesis revealed that the half-life of oligomerization was approximately 30 min, which correlated with the appearance of complexes consisting of M, HE, and S proteins, suggesting that oligomerization and/or conformational changes may be important for the S-M-HE protein complexes to form. Only HE dimers were found associated with the heterocomplexes consisting of all three proteins. S-M-HE protein complexes were detected prior to processing of the oligosaccharide chains on HE, indicating that these protein complexes formed in a premedial Golgi compartment before trimming of sugar chains. Transient coexpressions and double-labeling immunofluorescence demonstrated that HE and S proteins colocalized with M protein. This was further supported by coimmunoprecipitation of specific HE-M and S-M protein complexes from transfected cells, indicating that these proteins can form complexes in the absence of other viral proteins.


title: The risk of relapses in multiple sclerosis during systemic infections.

text: OBJECTIVES To assess the risk of multiple sclerosis (MS) relapses, MRI activity, and T cell responses during systemic infections (SI) in patients with MS. METHODS The authors prospectively studied 60 patients with MS. Twenty patients were evaluated with sequential MRI on initial visit, and 2 and 12 weeks later. Blood samples were collected at first infection symptom and 2, 5, 12, and 24 weeks later, and production of IL-4, IL-10, IL-12, IFN-gamma, TNFalpha, VLA-4, LFA-1, MMP-9, and MMP-2 were measured after infectious antigens (Ag) stimulation. RESULTS Increased risk of relapse and MRI activity were observed during SI. Numbers of IFN-gamma, TNFalpha, and IL-12 secreting cells, serum concentrations of MMP-9, and expression of VLA-4 and LFA-1 after PBMC viral or bacterial Ag stimulation were higher in samples collected during exacerbations associated to SI. Transwell analysis demonstrated that soluble factors produced during viral stimulation have little effect on myelin specific T cells activity. In contrast, PBMC viral stimulation in the presence of cognate myelin Ag induces maximal effector responses at 20 to 30 times lower than the Ag alone. CONCLUSIONS There was a significant association between systemic infections and risk of MS relapse, increased MRI activity, and T cells activation. Furthermore, infectious agents increased myelin specific T-cells sensitivity to cognate Ag.


title: Genetic diversity of avian infectious bronchitis virus isolates in Korea between 2003 and 2006.

text: Thirty-three field isolates of avian infectious bronchitis virus (IBV) were recovered from commercial chicken flocks in Korea between 2003 and 2006 and were characterized phylogenetically by nucleotide sequence analysis of the IBV S1 gene hyper-variable region. Our phylogenetic analysis revealed that recent field isolates of IBV formed at least three distinct phylogenetic types, including K-I, K-II, and K-III. K-I type IBV consisted of indigenous, 13 IBV isolates which evolved from the Kr-EJ/95 strain and then separated into the lineages of type K-Ia and type K-Ib. K-II type IBV isolates (n = 19) were closely related to nephropathogenic IBV variants from China and Japan. The K-III type isolate (Kr/D064/05), first identified by this study, was closely related to enteric IBV variants from the Chinese strains that cause proventriculitis. Sequence comparisons showed amino acid differences of >27.5% between IBV types. The molecular epidemiologic characteristics of IBV field isolates are briefly discussed.


title: Dog leukocyte antigen-haploidentical stem cell allografts after anti-CD44 therapy and nonmyeloablative conditioning in a preclinical canine model.

text: BACKGROUND We previously described a reduced-intensity hematopoietic cell transplantation (HCT) regimen in dog leukocyte antigen (DLA)-haploidentical littermate recipients consisting of 450 cGy total body irradiation (TBI) and anti-CD44 monoclonal antibody (mAb) S5 before and mycophenolate mofetil (MMF)/cyclosporine (CSP) after HCT. METHODS We tested a nonmyeloablative regimen of mAb S5 and 200 cGy TBI with postgrafting MMF/CSP in 44 DLA-haploidentical recipients using eight different regimens. Ten dogs also received escalating doses of donor lymphocyte infusion (DLI) alone or with pentostatin to convert to complete donor chimerism. RESULTS All dogs achieved initial engraftment between one to two weeks after HCT with peripheral blood mononuclear cell (PBMC) donor chimerism ranging from 2% to 98% (median 37%) on day +35. Twenty-five (57%) dogs rejected their donor grafts at a median of seven (range; 1-19) weeks after HCT. Low levels of PBMC donor chimerism at three weeks predicted graft rejection. DLI neither facilitated conversion to full donor chimerism after HCT nor prevented rejection. Higher total nucleated cells, CD4+, CD8+, and CD14+ cell subset numbers in the PBMC graft were associated with stable full donor engraftment. Dogs given higher cell subset doses of infused PBMC achieved longer duration of mixed chimerism. CONCLUSIONS Nonmyeloablative conditioning with 200 cGy TBI and anti-CD44 mAb was sufficient for initial uniform engraftment across DLA haplotype-mismatched barriers. However, sustained donor engraftment was seen in only 43% of recipients. Graft composition and donor-dominant chimerism early after HCT may be the most important factors for sustained donor engraftment.


title: Fasting prior to cardiac catheterisation: a single-centre observational study.

text: AIM Previous generation contrast agents were associated with high rates of nausea, vomiting and risk of aspiration leading to recommendations to fast prior to the procedure. However, modern contrast agents are well tolerated with a low risk of aspiration. Our current guidelines recommend fasting four to six hours before elective and semi-urgent cardiac catheterisation despite a lack of evidence to support this. We sought to determine the duration and effects of fasting at our centre. METHODS A single-centre prospective observational study in patients undergoing elective cardiac catheterisation over a six-month period between 7 August 2017 to 7 February 2018 at Auckland City Hospital, New Zealand. RESULTS One thousand and thirty patients with a mean age of 66±12 years underwent catheterisation. Sixty-seven percent were male, 26% had diabetes, 72% had hypertension and 23% had stage 3 or worse chronic kidney disease. The mean duration of fasting was 11.6±4.9 hours with 80% fasting longer than recommended. One hundred and eight (48%) patients with documented chronic kidney disease did not receive recommended pre-hydration. The most common symptoms related to fasting were hunger (47 %), nausea (3.9%) and vomiting (0.8%). Hypertension (4.1%) and hyperglycaemia (0.8%) occurred due to missed medication. There were no reports of aspiration. CONCLUSION Most patients were fasted for significantly longer than recommended and pre-hydration was underutilised in patients at high risk of contrast-induced nephropathy. There were no episodes of aspiration with modern contrast agents. Further studies are required to evaluate the need for fasting prior to non-emergency cardiac catheterisation.


title: Studies of avian urolithiasis associated with an infectious bronchitis virus.

text: Avian urolithiasis syndrome was diagnosed in 14-to-25-week-old chickens from a multiple-age caged-layer complex housing more than 2.5 million chickens. Losses from this syndrome ranged from 0.5 to 1.0% per week. Seven-to-14-week-old pullets from this facility had multifocal renal tubular necrosis leading to interstitial fibrosis, tophus formation, and tubular dilation. A coronavirus was isolated in embryos inoculated with pooled samples of trachea, kidney, and cecal tonsil of 4-week-old pullets. This virus, identified as 85-209, was related to infectious bronchitis virus strain Florida 88 by hemagglutination-inhibition assay. Day-old specific-pathogen-free chicks were inoculated with fifth-embryo-passage amnioallantoic fluid containing this virus. These chicks developed histologic lesions of tracheitis at 5 to 7 days postinoculation. Half the chicks inoculated by eyedrop developed renal tubular necrosis after 7 days. Urolithiasis in the flock investigated was attributed to renal damage by this strain of infectious bronchitis virus occurring in 4-to-7-week-old pullets and progressing to segmental atrophy, hyperplasia, and ureteral stone formation in 14-to-25-week-old chickens.


title: Necrotizing fasciitis: case report and review of literature.

text: We report a case of necrotizing fasciitis of the lower limb. This medico-surgical emergency is a life-threatening invasive soft-tissue infection which primarily involves the fascia superficialis and rapidly extends along subcutaneous tissue with relative sparing of skin and underlying muscles. Clinical presentation includes fever, signs of systemic toxicity and pain out of proportion to clinical findings. Paucity of cutaneous findings early in the course of the disease makes diagnosis challenging. The confirmation of the diagnosis is often made after surgical debridement. Delay in diagnosis and/or treatment correlates with poor outcome, leading to sepsis and/or multiple organ failure. Radiologic studies including plain radiographs, CT-scan or MRI may help to diagnose necrotizing fasciitis. Prompt surgical debridement, intravenous antibiotics, fluids and electrolytes management and analgesia are mainstays of the therapy. Adjuvant treatments like clindamycin, hyperbaric oxygen therapy and intravenous immunoglobulins are discussed.


Effective use of the clinical microbiology laboratory for diagnosing diarrheal diseases.


title: Angiotensin-converting enzyme 2 (ACE2), but not ACE, is preferentially localized to the apical surface of polarized kidney cells.

text: Angiotensin-converting enzyme-2 (ACE2) is a homologue of angiotensin-I converting enzyme (ACE), the central enzyme of the renin-angiotensin system (RAS). ACE2 is abundant in human kidney and heart and has been implicated in renal and cardiac function through its ability to hydrolyze Angiotensin II. Although ACE2 and ACE are both type I integral membrane proteins and share 61% protein sequence similarity, they display distinct modes of enzyme action and tissue distribution. This study characterized ACE2 at the plasma membrane of non-polarized Chinese hamster ovary (CHO) cells and polarized Madin-Darby canine kidney (MDCKII) epithelial cells and compared its cellular localization to its related enzyme, ACE, using indirect immunofluorescence, cell-surface biotinylation, Western analysis, and enzyme activity assays. This study shows ACE2 and ACE are both cell-surface proteins distributed evenly to detergent-soluble regions of the plasma membrane in CHO cells. However, in polarized MDCKII cells under steady-state conditions the two enzymes are differentially expressed. ACE2 is localized predominantly to the apical surface ( approximately 92%) where it is proteolytically cleaved within its ectodomain to release a soluble form. Comparatively, ACE is present on both the apical ( approximately 55%) and basolateral membranes ( approximately 45%) where it is also secreted but differentially; the ectodomain cleavage of ACE is 2.5-fold greater from the apical surface than the basolateral surface. These studies suggest that both ACE2 and ACE are ectoenzymes that have distinct localization and secretion patterns that determine their role on the cell surface in kidney epithelium and in urine.


title: Exploring the genetics, ecology of SARS-CoV-2 and climatic factors as possible control strategies against COVID-19.

text: The world has been thrown into pandemonium due to the recent Coronavirus Disease-19 (COVID-19) pandemic. Early available clinical data have indicated that geriatric persons cum those with co-morbidity such as cardiovascular, metabolic and immunological disorders suffered severe form of COVID-19. All countries and territories of the world are currently exploring available strategies to control the pandemic with the hope to significantly minimize its morbidity and mortality rate. This present study critically reviewed available and latest research progress of the genetics and ecology of SARS-CoV-2, as well as the influence of climatic factors on the spread of COVID-19, and thus, discussed how these concepts could be harnessed for COVID-19 control and further scientific advancements in resolving the pandemic.


title: Inhibition of the alpha/beta interferon response by mouse hepatitis virus at multiple levels.

text: Mouse hepatitis virus (MHV) was used as a model to study the interaction of coronaviruses with the alpha/beta interferon (IFN-alpha/beta) response. While MHV strain A59 appeared to induce IFN-beta gene transcription and low levels of nuclear translocation of the IFN-beta transcription factor interferon regulatory factor 3 (IRF-3), MHV did not induce IFN-beta protein production during the course of infection in L2 mouse fibroblast cells. In addition, MHV was able to significantly decrease the level of IFN-beta protein induced by both Newcastle disease virus (NDV) and Sendai virus infections, without targeting it for proteasomal degradation and without altering the nuclear translocation of IRF-3 or IFN-beta mRNA production or stability. These results indicate that MHV infection causes an inhibition of IFN-beta production at a posttranscriptional level, without altering RNA or protein stability. In contrast, MHV induced IFN-beta mRNA and protein production in the brains of infected animals, suggesting that the inhibitory mechanisms observed in vitro are not enough to prevent IFN-alpha/beta production in vivo. Furthermore, MHV replication is highly resistant to IFN-alpha/beta action, as indicated by unimpaired MHV replication in L2 cells pretreated with IFN-beta. However, when L2 cells were coinfected with MHV and NDV in the presence of IFN-beta, NDV, but not MHV, replication was inhibited. Thus, rather than disarming the antiviral activity induced by IFN-beta pretreatment completely, MHV may be inherently resistant to some aspects of the antiviral state induced by IFN-beta. These findings show that MHV employs unique strategies to circumvent the IFN-alpha/beta response at multiple steps.


The peritoneum.


title: Canine viral enteritis: prevalence of parvo-, corona- and rotavirus infections in dogs in the Netherlands.

text: After a brief review of the present knowledge about canine viral enteritis, the role played by parvoviral, coronaviral and rotaviral infections in contagious diarrhoea in dogs in the Netherlands is discussed. For this purpose a serologic survey, pathologic findings in dogs, and the demonstration of parvoviral antigen with an immunofluorescence test and with a newly developed haemadsorption-elution-haemagglutination assay (HEHA) are presented. It is concluded that infections with canine parvovirus, coronavirus and rotavirus appear widespread among dog populations in the Netherlands.


title: When ethical reform became law: the constitutional concerns raised by recent legislation in Taiwan.

text: In an effort at ethical reform, Taiwan recently revised the Hospice Palliative Care Law authorising family members or physicians to make surrogate decisions to discontinue life-sustaining treatment if an incompetent terminally ill patient did not express their wishes while still competent. In particular, Article 7 of the new law authorises the palliative care team, namely the physicians, to act as sole decision-makers on behalf of the incompetent terminally ill patient's best interests if no family member is available. However, the law fails to provide guidance as to what constitutes the patient's best interests or what specific procedures the treating physicians should follow, and so has raised constitutional concerns. It may be difficult to translate ethical reform into law but it is not impossible if essential requirements are carefully followed. First, there must be substantial nexus between the purpose of the statute and the measures provided under the statute. Second, advocates need to convince the public that futility or waste has amounted to a public health emergency so as to justify lower procedural requirements. Third, a remedy or compensation should be available if the surrogate decisions have not been appropriately made. Fourth,minimum procedural safeguards are necessary even though the statute is intended to reduce the procedural burdens of making surrogate decisions on behalf of incompetent patients who lack family members and did not express their wishes while still competent.


title: [The research and development of new diagnostic gene chips for SARS coronavirus].

text: The loci of cDNA sequences for valid diagnosis have been identified through the selection of the genome of SARS coronaries. The gene chips for diagnosing such virus have been developed, based on our own-developed technology for manufacturing and application of gene chips. The diagnoses given by such gene chips were consistent well with the reports of clinical laboratories (94.29%) and the sensitivity reached 10(-2)/ml virus molecules. This method is well suited for the clinical use in SARS coronaries diagnosis.


title: Severe acute respiratory syndrome and its impact on professionalism: qualitative study of physicians' behaviour during an emerging healthcare crisis.

text: OBJECTIVE To explore issues of medical professionalism in the context of severe acute respiratory syndrome (SARS), a new emerging health threat. DESIGN Qualitative interviews analysed with grounded theory methodology. SETTING University hospitals in Toronto, Canada, during the SARS outbreak in 2003. PARTICIPANTS 14 staff physicians from divisions of infectious diseases, general internal medicine, and critical care medicine. RESULTS Of 14 attending physicians, four became ill during the outbreak. Participants described their experiences during the outbreak and highlighted several themes about values inherent to medical professionalism that arose during this crisis including the balance between care of patients and accepted personal risk, confidentiality, appropriate interactions between physicians and patients, ethical research conduct, and role modelling of professionalism for junior doctors. CONCLUSION Despite concerns raised by professional societies about the erosion of professionalism, participants in this study amply demonstrated the necessary qualities during the recent healthcare crisis. However, there were several examples of strained professional behaviour witnessed by the participants and these examples highlight aspects of medical professionalism that medical educators and professional organisations should address in the future, including the balance between personal safety and duty of care.


title: Pathogenicity and sequence analysis studies suggest potential role of gene 3 in virulence of swine enteric and respiratory coronaviruses.

text: Coronaviruses have been commonly associated with enteric and respiratory diseases. Two of the swine coronaviruses, namely transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV), have been extensively studied. TGEV replicates in both the enteric and respiratory tracts and causes enteric disease, whereas, PRCV replicates in the respiratory tract with limited to no replication in the enteric tract. We have isolated PRCV from swine herds with respiratory disease and have reproduced moderate pneumonia in gnotobiotic and conventionally reared pigs with two of the PRCV isolates. We have also identified two PRCV isolates with low virulence. One consistent difference that we have observed between PRCV isolates of different pathogenicities is in gene 3. The gene 3 is intact in the two virulent PRCV isolates, whereas gene 3 is altered in the two low virulence isolates. A similar observation has been reported for TGEV as a nonpathogenic TGEV mutant with a small plaque morphology had a deletion in gene 3. We have also observed that one of the low virulence PRCV isolates, IA 1894, which has a deletion in gene 3, replicates poorly in cell cultures. Collectively these studies suggest that gene 3 may be an important determinant for in vivo virulence and in vitro replication of coronaviruses.


Letter to the editor: There is a need to consider all respiratory viruses in suspected mumps cases.


General concepts for PCR primer design.


title: Genetic susceptibility to otitis media.

text: PURPOSE OF REVIEW Otitis media is a disease that is prevalent in the pediatric population, and recent twin and triplet studies have confirmed that there is a strong genetic component to susceptibility. Here, we summarize the status of current efforts to identify the specific genes underlying otitis media susceptibility and presentation. RECENT FINDINGS Recent studies have focused on identifying candidate genes. For example, IFN-gamma polymorphisms, allotypes of the leukocyte IgG (FcgammaR) receptors and certain haplotypes of surfactant genes were linked in pilot studies to otitis media susceptibility. The pattern of gene expression during an episode of otitis media is also being elucidated with the overall goal of providing clues as to which of these modulated genes are polymorphic and thus potentially capable of affecting otitis media susceptibility. Mucin and cathepsin protease genes were shown to exhibit these characteristics. SUMMARY In addition to the simple searches for linkages between known genes and otitis media, work is progressing within the context of genome-wide linkage studies. These efforts promise to answer some of the many questions remaining in otitis media susceptibility and pathogenesis.


title: Management of respiratory viral infections in hematopoietic cell transplant recipients.

text: Advances in stem cell transplantation procedures and the overall improvement in the clinical management of hematopoietic cell transplant (HCT) recipients over the past 2 decades have led to an increase in survival duration, in part owing to better strategies for prevention and treatment of post-transplant complications, including opportunistic infections. However, post-HCT infections remain a concern for HCT recipients, particularly infections caused by community respiratory viruses (CRVs), which can lead to significant morbidity and mortality. These viruses can potentially cause lower respiratory tract illness, which is associated with a higher mortality rate among HCT recipients. Clinical management of CRV infections in HCT recipients includes supportive care and antiviral therapy, especially in high-risk individuals, when available. Directed antiviral therapy is only available for influenza infections, where successful use of neuraminidase inhibitors (oseltamivir or zanamivir) and/or M2 inhibitors (amantadine or rimantadine) has been reported. Data on the successful use of ribavirin, with or without immunomodulators, for respiratory syncytial virus infections in HCT recipients has emerged over the past 2 decades but is still controversial at best because of a lack of randomized controlled trials. Because of the lack of directed antiviral therapy for most of these viruses, prevention should be emphasized for healthcare workers, patients, family, and friends and should include the promotion of the licensed inactivated influenza vaccine for HCT recipients, when indicated. In this review, we discuss the clinical management of respiratory viruses in this special patient population, focusing on commercially available antivirals, adjuvant therapy, and novel drugs under investigation, as well as on available means for prevention.


title: Nucleocapsid-independent specific viral RNA packaging via viral envelope protein and viral RNA signal.

text: For any of the enveloped RNA viruses studied to date, recognition of a specific RNA packaging signal by the virus's nucleocapsid (N) protein is the first step described in the process of viral RNA packaging. In the murine coronavirus a selective interaction between the viral transmembrane envelope protein M and the viral ribonucleoprotein complex, composed of N protein and viral RNA containing a short cis-acting RNA element, the packaging signal, determines the selective RNA packaging into virus particles. In this report we show that expressed coronavirus envelope protein M specifically interacted with coexpressed noncoronavirus RNA transcripts containing the short viral packaging signal in the absence of coronavirus N protein. Furthermore, this M protein-packaging signal interaction led to specific packaging of the packaging signal-containing RNA transcripts into coronavirus-like particles in the absence of N protein. These findings not only highlight a novel RNA packaging mechanism for an enveloped virus, where the specific RNA packaging can occur without the core or N protein, but also point to a new, biologically important general model of precise and selective interaction between transmembrane proteins and specific RNA elements.


title: Molecular dissection of ø29 scaffolding protein function in an in vitro assembly system.

text: An in vitro assembly system was developed to study prolate capsid assembly of phage ø29 biochemically, and to identify regions of scaffolding protein required for its functions. The crowding agent polyethylene glycol can induce bacteriophage ø29 monomeric capsid protein and dimeric scaffolding protein to co-assemble to form particles which have the same geometry as either prolate T=3 Q=5 procapsids formed in vivo or previously observed isometric particles. The formation of particles is a scaffolding-dependent reaction. The balance between the fidelity and efficiency of assembly is controlled by the concentration of crowding agent and temperature. The assembly process is salt sensitive, suggesting that the interactions between the scaffolding and coat proteins are electrostatic. Three N-terminal ø29 scaffolding protein deletion mutants, Delta 1-9, Delta 1-15 and Delta 1-22, abolish the assembly activity. Circular dichroism spectra indicate that these N-terminal deletions are accompanied by a loss of helicity. The inability of these proteins to dimerize suggests that the N-terminal region of the scaffolding protein contributes to the dimer interface and maintains the structural integrity of the dimeric protein. Two C-terminal scaffolding protein deletion mutants, Delta 79-97 and Delta 62-97, also fail to promote assembly. However, the secondary structure and the dimerization ability of these mutants are unchanged relative to wild-type, which suggests that the C terminus is the likely site of interaction with the capsid protein.


title: State-specific influenza vaccination coverage among adults aged > or =18 years--United States, 2003-04 and 2005-06 influenza seasons.

text: Influenza epidemics occur seasonally and result in substantial morbidity and mortality among adults in the United States. Adult groups included in the 2007 Advisory Committee on Immunization Practices (ACIP) recommendation for annual influenza vaccination are persons aged 18-49 years with high-risk conditions (i.e., conditions associated with an increased risk for complications from influenza), persons aged > or =50 years, health-care personnel, and others who are household contacts or caregivers of persons at high risk (e.g., persons with high-risk conditions or children aged < or =59 months). In addition, adults who want to reduce the risk for becoming ill with influenza or of transmitting influenza to others should be vaccinated. Healthy People 2010 (HP2010) objectives include increasing vaccination levels to 90% for adults aged > or =65 years (objective 14-29a) and 60% for persons aged 18-64 years who have one or more high-risk conditions (objective 14-29c). From the 1992-93 through 2003-04 influenza seasons, seasonal influenza vaccination coverage estimates (based on Behavioral Risk Factor Surveillance System [BRFSS] data) among adults aged > or =65 years trended upward, except for three seasons (1997-98, 1999-00, and 2000-01) when no increases occurred (Figure). To evaluate recent state-specific progress toward the HP2010 objectives, CDC compared data from the 2004 and 2006 BRFSS surveys, which reflected vaccinations received during the 2003-04 and 2005-06 influenza seasons; data from the 2004-05 influenza season, which have been published previously, were not included in this comparison because that season was marked by a substantial shortage of influenza vaccine. This report describes the results of the analysis, which indicated that influenza vaccination coverage for the 2005-06 season did not return to levels observed before the vaccine shortage of 2004-05 and remained substantially below HP2010 targets. Comprehensive measures are needed to improve influenza vaccination coverage among adult populations in the United States, including increasing adoption of recommended adult immunization practices by health-care providers, raising public awareness about influenza vaccination, vaccinating throughout the influenza season, and ensuring stable supplies of readily available vaccine.


title: The NS1 protein of tick-borne encephalitis virus forms multimeric species upon secretion from the host cell.

text: Flaviviruses elicit a humoral immune response to two virus-encoded, membrane-associated glycoproteins. One is the major virion surface envelope protein (E), which is recognized by antibody, whereas the other is a secreted, heavily glycosylated non-structural protein (NS1). Inoculation with either protein can give rise to a protective immune response, as can the passive transfer of E and NS1 monospecific monoclonal antibodies. Experiments reported here demonstrate that the secreted form of NS1, whether from cells infected with tick-borne encephalitis virus (TBEV) or from cells infected with a defective recombinant adenovirus containing the NS1 gene, occurs chiefly as a pentamer or hexamer and occasionally as a decamer or dodecamer. Intracellular forms of this protein however occur only as dimers. The higher M(r) forms secreted from the cell are exquisitely sensitive to detergent, suggesting they are held together by hydrophobic bonds. Both intracellular and extracellular forms of the dimer can be dissociated by heat, but at different temperatures. Unlike similar proteins from mosquito-borne viruses. NS1 from TBEV-infected cells cannot be dissociated at ambient temperatures by extremes of pH. Studies on the antigenic structure of this protein show it to have several highly conserved epitopes, confirming similar earlier conclusions from amino acid sequence analyses.


The Public Health Implications of Religious Exemptions: A Balance Between Public Safety and Personal Choice, or Religion Gone Too Far?


Background paper. Progress towards a coronavirus recombinant DNA vaccine.


Biomarker discovery and analysis platform: application to Alzheimer's disease.


title: [Ethical dilemmas in caring for patients with SARS].

text: This paper explored experiences caring for patients with severe acute respiratory syndrome (SARS), with a focus on the suffering between the "private" and "professional" selves, in order to describe the psychological impact and ethical practice of SARS events as related to nursing professionals. Eight small group interviews were conducted with a total of twenty-one nurses with experience caring for SARS patients. Group interviews were audio-taped and transcribed for analysis. Data, analyzed using the constant comparative method, revealed the following three ethical dilemmas: suffering in care giving, response to the calling, and distresses involved in decision making. In this study, the authors further addressed relationships between suffering and ethical practice, the calling and a nurse's professional responsibilities, and transcendence in intersubjectivity. The authors hope that the results of this study will lead to a new understanding of ethical practices nursing and help nurses develop reflected care that promotes personal and professional growth.


title: Role of subgenomic minus-strand RNA in coronavirus replication.

text: Coronavirus subgenomic minus-strand RNAs (negative-strand copies of the 3' coterminal subgenomic mRNAs) probably function in mRNA amplification by serving as templates for transcription from internal (intergenic) promoters, rather than by faithful (full-length) mRNA replication.


title: Clinical course of community-acquired respiratory syncytial virus pneumonia in newborns hospitalized in neonatal intensive care unit.

text: INTRODUCTION Respiratory syncytial virus (RSV) is a major cause of pneumonia in infants worldwide. We aimed to evaluate the clinical course of community-acquired RSV pneumonia in newborns hospitalized in neonatal intensive care unit. MATERIALS AND METHODS All the newborns diagnosed as pneumonia were prospectively evaluated for RSV infection between November 2010 and April 2011. Fifty-four specimens of nasopharyngeal secretions were tested in parallel with the RAT and the multiplex real time polymerase chain reaction (RT- PCR). Downes' score was used to assess the disease severity in patients with pneumonia. RESULTS RAT has a sensitivity of 90% and a specificity of 78.5%, as the PCR technique target assay. Four of the patients with RSV pneumonia had secundum atrial septal defect (ASD) and all of four needed mechanical ventilation support. The first day Downes' score was positively correlated with time of intravenous fluid requirement (p= 0.001, r= 0.48), total oxygen need (p= 0.000, r= 0.63), and re-enteral feeding (p= 0.001, r= 0.46). Blood pH (p= 0.031, r= 0.46) were negatively correlated with Downes' score. The second day Downes' score was higher in patients with ASD than those of without ASD (3.8 ± 2.6 vs. 2 ± 1.1, p= 0.01). The most possible risk factor for longer hospital stay was the higher second day Downes' score (p= 0.02 OR: 1.9, CI 95% (1.1-3.2). All infants were discharged from hospital in a good health. CONCLUSION RAT is sensitive and specific in detecting RSV infections in newborns. Physicians may use Downes' score for evaluation of disease severity in infants with RSV pneumonia. In these patients, ASD has increased the disease severity.


Further identification and characterization of products processed from the coronavirus avian infectious bronchitis virus (IBV) 1a polyprotein by the 3C-like proteinase.


title: Impact of severe respiratory syndrome on anxiety levels of front-line health care workers.

text: OBJECTIVE To identify anxiety levels among front-line health care workers during the 2003 severe acute respiratory syndrome outbreak. DESIGN Questionnaire survey. SETTING Regional hospital, Hong Kong. PARTICIPANTS All hospital staff were given a questionnaire; administrative staff who had not had any patient contact served as controls. MAIN OUTCOME MEASURES Levels of contact with patients who had severe acute respiratory syndrome were measured and correlated with anxiety levels as determined by the State-Trait Anxiety Inventory. RESULTS Of 4252 questionnaires distributed between May and June 2003, 2040 (48.0%) were returned and 1926 (45.3%) were valid for analysis. Overall, 534 (27.7%) respondents had had contact with patients with severe acute respiratory syndrome. Anxiety scores ranged from 20 to 80, and mean (standard deviation) scores were higher among staff who had had contact with patients with severe acute respiratory syndrome than among those who had not (52.6 [10.5] versus 49.8 [10.1], respectively; P<0.01). Mean anxiety levels were higher among workmen, health care assistants, and nurses than among administrative staff controls or doctors (P<0.01). Anxiety scores were correlated with burnout scores (Pearson's correlation coefficient, 0.52-0.59) and with discomfort from wearing protective gear (0.21-0.32). CONCLUSION Severe acute respiratory syndrome has likely stressed the public health care system. Prediction and early identification of adverse factors in a crisis situation would allow early implementation of interventions to reduce and counteract the impact of this stress.


title: Applicability of a real-time quantitative PCR assay for diagnosis of respiratory syncytial virus infection in immunocompromised adults.

text: Respiratory syncytial virus (RSV) accounts for the majority of respiratory virus infections, producing high mortality rates in immunocompromised patients with hematologic malignancies. The available methods for the rapid detection of RSV by antigen detection or PCR either lack sensitivity, require complex laboratory manipulation, or have not been evaluated in this patient population. To assess the applicability of a TaqMan-based real-time PCR technique for the detection of RSV A and B in immunocompromised adults, we developed a rapid, sensitive detection method that simultaneously detects RSV A and B and can be applied in routine diagnostics. The specificity of the assay was assessed using a panel of reference strains of other respiratory viruses and RSV. Electron microscopy-counted stocks of RSV A and B were used to develop a quantitative PCR format. Eleven copies of viral RNA could be detected for RSV A strain Long, and 14 copies could be detected for RSV B strain 9320, corresponding to 50% tissue culture infective doses of 0.86 and 0.34, respectively. The assay was evaluated on 411 combined nose and throat swabs derived from immunocompromised adults with or without signs of respiratory tract infection. The diagnostic efficacy of the TaqMan PCR determined on the clinical samples showed that this real-time PCR technique was substantially more sensitive than the combination of conventional viral culture and shell vial culture. None of the clinical specimens derived from patients without signs of respiratory illness were found to be positive for RSV by real-time TaqMan PCR.


title: Psychosomatic medicine and biodefense preparedness--a new role for the American Psychosomatic Society.

text: Biodefense preparations in the United States have focused mostly on improving biosurveillance and hospital surge capacity in the event of an outbreak or a weapons of mass destruction (WMD) event. However, what if an invisible bioweapon or dirty bomb was released in a major population center, or if avian flu took hold with sustained human to human transmission? Suddenly, we need to combine efforts from psychosomatic medicine and general medicine with public health practice to triage nonexposed patients with somatic symptoms from those with medical sequelae resulting from hazardous exposures. This would better enable the limited acute care resources to be directed to those most in need of urgent medical care. Furthermore, psychosomatic medicine experts are potentially important players in biodefense planning related to risk communication and health education strategies in a WMD scenario or outbreak in which individuals must make informed choices about their need for immediate medical attention.


title: Feline calicivirus replication: requirement for polypyrimidine tract-binding protein is temperature-dependent.

text: The interaction of host-cell nucleic acid-binding proteins with the genomes of positive-stranded RNA viruses is known to play a role in the translation and replication of many viruses. To date, however, the characterization of similar interactions with the genomes of members of the family Caliciviridae has been limited to in vitro binding analysis. In this study, Feline calicivirus (FCV) has been used as a model system to identify and characterize the role of host-cell factors that interact with the viral RNA. It was demonstrated that polypyrimidine tract-binding protein (PTB) interacts specifically with the 5' sequences of the FCV genomic and subgenomic RNAs. Using RNA interference it was shown that PTB is required for efficient FCV replication in a temperature-dependent manner. siRNA-mediated knockdown of PTB resulted in a 15- to 100-fold reduction in virus titre, as well as a concomitant reduction in viral RNA and protein synthesis at 32 degrees C. In addition, virus-induced cytopathic effect was significantly delayed as a result of an siRNA-mediated reduction in PTB levels. A role for PTB in the calicivirus life cycle was more apparent at temperatures above and below 37 degrees C, fitting with the hypothesis that PTB functions as an RNA chaperone, potentially aiding the folding of RNA into functional structures. This is the first functional demonstration of a host-cell protein interacting with a calicivirus RNA.


title: The emerging role of RNA interference in the design of novel therapeutics in oncology.

text: RNA interference (RNAi) has emerged as the tool of choice for studying gene function. Dubbed the "breakthrough of the year" in 2002 by the journal Science, RNAi is a naturally occurring host defense mechanism that mediates the sequence-specific degradation of target mRNA transcripts and their protein products. The specificity of RNAi makes it an ideal tool for targeted therapeutics against unique fusion oncogene sequences. RNAi may also be effective against viral-mediated oncogenesis and has the potential to enhance tumor sensitivities to existing chemotherapy. The current interest in the success of RNAi-based therapies will depend on the delivery systems that protect the silencing apparatus from endogenous nucleases, sustain tissue-specific expression of the small-interfering RNAs, and prevent the activation of a destructive nonspecific host immune response.


title: Identification of antigenic sites mediating antibody-dependent enhancement of feline infectious peritonitis virus infectivity.

text: We have previously demonstrated antibody-dependent enhancement of feline infectious peritonitis virus (FIPV) infection of macrophages using both virus-specific antisera and monoclonal antibodies (MAbs) to the spike (S) protein of FIPV. To increase our understanding of this phenomenon, six representative MAbs from a previously documented group of 12 enhancing MAbs were used to identify epitopes that mediate antibody-dependent enhancement of FIPV infectivity. Analysis of the results of kinetics-based competitive ELISA (K-cELISA) among these six enhancing MAbs grouped the epitopes into two clusters. Because transmissible gastroenteritis virus (TGEV) and FIPV are so closely related antigenically, we also conducted K-cELISA experiments between the FIPV MAbs and TGEV S protein-specific MAbs for which the epitopes had previously been mapped to specific sites on the TGEV S protein. Results of these assays indicated that the two FIPV epitope clusters are homologues of the previously defined TGEV S protein sites A and E/F. In addition, two TGEV S protein-specific MAbs also induced antibody-dependent enhancement of FIPV infection of macrophages. This functional cross-reactivity provides further support for the close antigenic relationship between FIPV and TGEV. Our results provide a preliminary localization of several enhancing epitopes within the amino acid sequence of the FIPV S protein.


title: Cosmetic Surgery-Use of 3mm Ports and Reduced-Port Techniques for Gynecologic Surgery.

text: As innovation continues to evolve in gynecologic surgery, new "cosmetic" techniques are continually being developed. Minimally invasive surgery offers clear benefits over open surgery, such as decreased postoperative pain, infection, length of hospital stay, and time out of work. It is now possible to perform many laparoscopic gynecologic surgeries with minimal to no visible scars. This article reviews the use of 3mm ports and several reduced-port and single-port techniques for use in gynecologic surgery as well as vaginal natural orifice transluminal endoscopic surgery (vNOTES). We also discuss the advantages and challenges that physicians face regarding the adoption of these procedures into general practice. Lastly, we present our novel economic adaption in approach to reduced-port laparoscopic bilateral salpingectomy versus traditional single-site laparoscopic bilateral salpingectomy. In conclusion, minimally invasive approaches to gynecological surgery are considered feasible and safe.


title: A method to increase efficiency in testing pooled field-collected mosquitoes.

text: Testing field-caught mosquito collections can result in thousands of pools, and testing pools of 50 mosquitoes each can be both time consuming and cost prohibitive. Consequently, we have developed an alternative approach to testing mosquito pools for arboviruses, utilizing a superpool strategy. When mosquito samples are processed for extraction of viral RNA and subsequent virus testing via quantitative real-time polymerase chain reaction, each pool is tested individually. Using the method described here, 0.025 ml from each of 10 pools is combined into a superpool for RNA extraction and testing. When a virus-positive superpool sample is found, each of the original 10 pools that constitute this sample is tested individually in order to find the specific positive sample. By retesting the original samples after the initial superpool screen, we are still able to obtain reliable estimates for minimum infection rates or maximum likelihood estimations. To test this principle, we created controlled mosquito pools of known titer and subjected them to our superpool process. We were able to detect our entire range of laboratory-created pools as being West Nile virus (WNV) positive. In 2005, field surveillance efforts from our laboratory resulted in over 4,000 mosquito pools tested, with 8 resulting WNV-positive samples. We found that all of these field samples were detected as WNV positive using the superpool method and contained calculated virus titers from < 0.1 to 4.1 log10 plaque-forming units/ml WNV, indicating that the limit of superpool detection of WNV is below this point. These results reveal that the superpool method could be accurately used to detect WNV in field-collected specimens.


Rising rates of HIV infection.


title: Planning for avian influenza.

text: Avian influenza, or influenza A (H5N1), has 3 of the 4 properties necessary to cause a serious pandemic: It can infect people, nearly all people are immunologically naive, and it is highly lethal. The Achilles heel of the virus is the lack of sustained human-human transmission. Fortunately, among the 124 cases reported through 30 May 2006, nearly all were acquired by direct contact with poultry. Unfortunately, the capability for efficient human-human transmission requires only a single mutation by a virus that is notoriously genetically unstable, hence the need for a new vaccine each year for seasonal influenza. Influenza A (H5N1) is being compared to another avian strain, the agent of the "Spanish flu" of 1918-1919, which traversed the world in 3 months and caused an estimated 50 million deaths. The question is, are we ready for this type of pandemic? The answer is probably no. The main problems are the lack of an effective vaccine, very poor surge capacity, a health care system that could not accommodate even a modest pandemic, and erratic regional planning. It's time to get ready, and in the process be ready for bioterrorism, natural disasters, and epidemics of other infectious diseases.


title: Detection of gill-associated virus (GAV) by in situ hybridization during acute and chronic infections of Penaeus monodon and P. esculentus.

text: Chronic and acute gill-associated virus (GAV) infections were examined by in situ hybridization (ISH) using a DNA probe targeting a 779 nucleotide region of the ORF1b-gene. Chronic GAV infections were observed in healthy Penaeus monodon collected from farms and healthy P. esculentus surviving experimental infection. During chronic-phase infections in both species, GAV was detected only in partitioned foci of cells with hypertrophied nuclei (spheroids) within the lymphoid organ. Acute-phase infections were observed in moribund P. monodon and P. esculentus infected experimentally with a high dose of GAV, and in moribund P. monodon collected from farms during outbreaks of disease. During acute experimental infections in P. monodon, ISH detected GAV throughout the lymphoid organ, in gills and in connective tissues throughout the cephalothorax. In moribund P. monodon collected from natural outbreaks of disease, GAV was also detected in the gills and in connective tissues of the cephalothorax, but the distribution of virus within the lymphoid organ varied. In acutely infected P. esculentus, GAV was detected in connective tissues, but was restricted to the inner stromal matrix cells and endothelial cells of intact lymphoid organ tubules. The tissue distribution of GAV identified by ISH suggests that shrimp are able to control and maintain chronic asymptomatic infection by a process involving lymphoid organ spheroids. Acute phase infections and the development of disease appear to be dose-related and involve the systemic distribution of virus in connective tissues throughout the cephalothorax.


title: Dispersal of a blow fly, Calliphora nigribarbis, in relation to the dissemination of highly pathogenic avian influenza virus.

text: A mark-release-recapture study of the dispersal ability of blow flies, Calliphora nigribarbis, was conducted in Ikumo-Naka, Ato Town, Yamaguchi Prefecture, Japan in December 2004. A location where a fatal avian influenza outbreak had occurred 1 year previously was selected for the present study. A total of 3,884 C. nigribarbis were collected, 1,915 of which were marked and released from 4 different collection sites during 2 successive days. The recapture rate of the released C. nigribarbis ranged from 0.014 to 0.029 among the collection sites, and the overall recapture rate was calculated as 0.022. Based on the distance between the released site and the recaptured site, the dispersal rate of C. nigribarbis was estimated as 256 m/h on the 1st day and 179 m/h on the 2nd day of the experiment, and the maximum dispersal rate observed in this study was estimated as 500 m/h. Taking into account the active period of C. nigribarbis on a fine day (7 h/day), the distance traveled by C. nigribarbis within a day was estimated as 1,789 and 1,250 m/day on average for the 1st and 2nd days, respectively, and the maximum distance was 3,500 m/day. These results suggest that C. nigribarbis could play a role in the mechanical dissemination of avian influenza virus and spread of the outbreak in Japan.


title: Detection of group B rotaviruses in fecal samples from diarrheic calves and adult cows and characterization of their VP7 genes.

text: Groups A, B, and C rotaviruses have been identified in cattle. Group B rotaviruses are associated with sporadic cases of diarrhea in calves and adult cows. From diagnostic submissions to our laboratory, 90 fecal samples from cases of calf diarrhea, 81 fecal samples from cases of adult cow diarrhea (winter dysentery), and 20 fecal samples from case control normal adult cows were tested for group B rotaviruses by polyacrylamide gel electrophoresis (PAGE), and reverse transcription (RT)-PCR (targeting 279 bp of the VP7 gene). In addition, 53 fecal samples from diarrheic adult cows were tested for group B rotaviruses by immune electron microscopy (IEM). By RT-PCR, five samples from calves were group B rotavirus positive (5.6%). Fifteen samples from adult cows with diarrhea were group B rotavirus positive (18.5%), and none of the control fecal samples from normal cows were positive for group B rotaviruses. By PAGE, one calf sample (RT-PCR positive) was group B rotavirus positive (short electropherotype), but none of the adult cow samples were positive for group B rotaviruses. By IEM, 5 (9.4%) of the 53 fecal samples from diarrheic adult cows were group B positive (all were also RT-PCR positive). The VP7 genes of three strains (WD653 from an adult cow and the ATI and Mebus calf strains) were sequenced. The VP7 genes from the three bovine strains showed high (over 90%) nucleotide and deduced amino acid homologies, but lower homologies (48 to 61%) were seen between these genes and the genes from rodent (IDIR) and human (ADRV) group B rotaviruses. Although there were some differences of degree, all inoculated gnotobiotic calves (n = 6) showed abnormal feces between 1 and 3 days after inoculation with each of three strains of group B bovine rotaviruses, and group B rotaviruse, were detected in the feces for up to 2 weeks by RT-PCR but for shorter periods by PAGE or IEM.


title: Respiratory infections: respiratory infections in immigrants to the United States.

text: Immigration has brought millions of individuals into the United States over the past decade. The Centers for Disease Control and Prevention and the US Public Health Service are charged with ensuring that immigrants who enter do not pose a public health risk. Health examinations and immunization regimens are required for individuals wishing to live in the United States. Many immigrants and refugees are exposed to communicable diseases not routinely encountered in the United States These include helminthic infections, tuberculosis, malaria, and other infections. Zoonotic infections, such as influenza A, and novel coronavirus infections also are of increasing concern because of population mobility.


title: [Influence of concanavalin A on the attachment of the coronavirus of transmissible gastroenteritis in cell cultures].

text: Effect of Concanavalin (ConA) on attachment of three strains of Transmissible Gastroenteritis coronavirus (TGE) of swine was investigated in cell culture. Whatever the virus strain, reduction of virus plaques number is observed when viral suspension is incubated with cells together or after addition of ConA. Intensity of inhibition is related with ConA concentration. ConA treatment of preinfected cell cultures has no effect on plaque formation. Addition of alpha-methyl-D-mannoside inhibits the ConA activity. Treatment of cells with metaperiodate has no effect on plaque formation and ConA activity. Our results suggest that ConA inhibits formation of plaques by TGE coronavirus.


title: Mental health services in primary care.

text: In the UK, only 13% of people with long-term mental health problems are in employment, compared with 35% generally of people with a disability (Royal College of General Practitioners, 2005). Nearly 2.6 million individuals receive incapacity benefit and/or severe disability allowance and, of these, close to 1 million are claiming incapacity benefit due to mental ill health. The management of this enormous number of people - providing support to them and helping them get back into employment - is an issue that cannot be addressed adequately by our specialist mental health services. Accordingly, other models of service delivery need to be considered. The three thematic papers in this issue look at this issue from the perspective of three highly contrasting societies.


title: Epidemic and bioterrorism preparation among emergency medical services systems.

text: INTRODUCTION The purpose of this research was to determine the preparedness of emergency medical services (EMS) agencies in one US state to cope with a massive epidemic event. METHODS Data were collected primarily through telephone interviews with EMS officials throughout the State. To provide a comparison, nine out-of-state emergency services agencies were invited to participate. RESULTS Emergency medical services agencies from nine of the 23 counties (39%) provided responses to some or all of the questions in the telephone survey. Seven of the nine out-of-state agencies provided responses to the survey. Most of the EMS agencies do not have broad, formal plans for response to large-scale bio-terrorist or pandemic events. CONCLUSIONS The findings indicate that EMS agencies in this state fundamentally are unprepared for a large-scale bioterrorism or pandemic event. The few existing plans rely heavily on mutual aid from agencies that may be incapable of providing such aid. Therefore, EMS agencies must be prepared to manage a response to these incidents without assistance from any agencies outside of their local community. In order to accomplish this, they must begin planning and develop close working relationships with public health, healthcare, and elected officials within their local communities.


Rotavirus and coronavirus associated diarrhoea in domestic animals.


title: Host factors in the replication of positive-strand RNA viruses.

text: Viruses are obligate, intracellular parasites that depend on host cells for successful propagation. Upon infection of host cells, positivestrand RNA viruses exploit and hijack cellular machinery and reprogram these cells into viral "factories" through various protein-protein, protein- RNA, and protein-lipid interactions. The molecular interplay between host factors and invading viruses is a continuous process throughout the entire viral life cycle and determines virus host range and viral pathogenesis, as well as driving viral evolution. Studies of host factors have contributed insights into their normal cellular functions and helped identify attractive targets for antiviral drug development. With the development of high throughput screening, functional genomics, and proteomics technologies, host factors participating in viral life cycles have been identified rapidly in recent years. In this review, we summarize the recent advances in virus-host cell interactions in positive-strand RNA virus infections and focus on host factors that facilitate viral replication.


title: [Infections with the MERS coronavirus--for the present no threat to Europe].

text: In Saudi Arabia, a novel coronavirus named Middle East respiratory syndrome coronavirus (MERS-CoV) was isolated in 2012 from patients with severe respiratory symptoms. Up to now, more than 1600 MERS cases have been registered mainly in the Arabian Peninsula. MERS is usually accompanied with fever, cough, and shortness of breath. In many cases, pneumonia is observed. However, clinical features of MERS range from mild disease to acute respiratory distress syndrome and multiorgan failure resulting in death, especially in individuals with underlying comorbidities. To date, about one in three people died as a result of MERS. In Europe, MERS cases have only been registered in isolated travelers entering from the Middle East.


title: Genome of infectious bronchitis virus.

text: Techniques are described for the growth and rapid purification of the avian coronavirus infectious bronchitis virus (IBV). Purified IBV has a sedimentation coefficient of 320S and a buoyant density of 1.22 g/ml in sucrose-deuterium oxide equilibrium gradients. IBV RNA extracted by proteinase K in the presence of sodium dodecyl sulfate and further purified by phenol extraction and gradient centrifugation is single stranded and has a sedimentation coefficient of 64S, as determined by isokinetic gradient centrifugation. Analysis on sucrose gradients under both aqueous and denaturing conditions together with agarose gel electrophoresis in the presence of the chaotropic agent methylmercuric hydroxide gave a value of 8 X 10(6) for the moleclar weight of IBV RNA. This value was confirmed by RNase T1 fingerprinting, which also indicated that IBV RNA is haploid. No evidence was found of subunit structure in IBV RNA. From these results together with the recently reported observation that IBV RNA is infectious and contains a tract of polyadenylic acid (Lomniczi, J. Gen. Virol., in press), we conclude that the genome of the coronaviruses is a single continuous chain of about 23,000 mononucleotides that is of messenger polarity.


title: Fusogenic properties of uncleaved spike protein of murine coronavirus JHMV.

text: We have tested the fusogenic properties of cleaved and uncleaved spike (S) protein of murine coronavirus (MCV) JHMV variant cl-2 by expressing the S protein by recombinant vaccinia viruses (RVVs). The amino acid sequence of the putative cleavage site of cl-2 S protein, Arg-Arg-Ala-Arg-Arg, was replaced by Arg-Thr-Ala-Leu-Glu by in vitro mutagenesis of cl-2 S gene. The RVVs having cl-2 S gene [RVV t(+)] or mutated cl-2 S gene [RVV t(-)] were tested for their ability to induce fusion as well as cleavability in DBT cells. After inoculation with RVV t(+) onto DBT cells, the fusion formation was first observed at 8 h postinoculation (p.i.) and spread throughout the whole culture by 24 h. In cells infected with RVV t(-), fusion appeared by 2 h and most of cells were fused by 30 h p.i. The S protein and its cleavage products were detected in DBT cells expressing wild type S protein. However, no cleavage products of the S protein were detected in RVV t(-) infected cells producing mutated S protein, even though fusion was clearly visible. These results suggest that the cleavage event of JHMV-S protein of MCV is not a prerequisite for fusion formation, but that it enhances fusion.


Statement of ESMINT and ESNR regarding recent trials evaluating the endovascular treatment at the acute stage of ischemic stroke.


title: Recombinant herpesviruses as tools for the study of herpesvirus biology.

text: This article is a brief summary of efforts to generate mutant herpesviruses for investigating and assigning gene functions of herpesviruses in replication and pathogenesis. While a full review of all herpesviruses is beyond the scope of this review, we focused our attention on the prototype of the herpesvirus subfamily - herpes simplex virus and murine gammaherpesvirus that serves as an excellent animal model to study human gammaherpesvirus pathogenesis. Furthermore, our present knowledge of essential, non-essential, and common genes of herpesviruses as well as of accessory genes that are currently being studied with the help of the bacterial artificial chromosome (BAC) system will also be discussed. This system facilitates the analysis of herpesviral genes with potential for use in gene therapy or as anti-cancer therapeutics.


Outbreak of E. coli O157:H7 associated with bathing at a public beach in the Montreal-Centre region.


title: Clinical analysis of 45 patients with severe acute respiratory syndrome.

text: OBJECTIVE To explore the clinical and radiological features of severe acute respiratory syndrome (SARS). METHODS Analysis of the clinical presentation, chest radiographs, course of disease and features of severe pneumonia in 45 SARS patients diagnosed at our hospital in Beijing between April 5 - 20, 2003. Also included is a summation of the clinical features of SARS. RESULTS (1) SARS appears to have high infectivity; (2) the most common symptom is fever; (3) the count of leukocyte is normal or decreased; (4) most patients (35/45, 77.8%) had experienced a 24-hour fever prior to the abnormal chest X-ray changes which showed progression of pulmonary infiltrates within 48 hours in 71.1% (32/45) of the patients and, (5) the percentage of patients who developed severe pneumonia (24.4%) is higher than those who developed typical pneumonia. CONCLUSION SARS is a disease with high infectivity and has its own clinical and radiological features. Early recognition, prompt isolation, and appropriate therapy are the key to combat this infection.


title: Exploratory rigid laparoscopy in an African elephant (Loxodonta africana).

text: In March 2009, a 25-yr-old captive female African elephant (Loxodonta africana) underwent an exploratory laparoscopy after several weeks of diarrhea, submandibular and ventral edema, and swelling on medial and lateral aspects of all feet. Although there have been recent advances in laparoscopic vasectomies in free-ranging African elephants in South Africa utilizing specially designed rigid laparoscopes and insufflation devices, this was the first attempt at using these same techniques for an exploratory purpose. The elephant was sedated in a static restraint chute and remained standing for the duration of the procedure. Laparoscopy provided visibility of the dorsal abdomen, enabled collection of reproductive tract biopsies and peritoneal fluid samples, and allowed for instillation of antibiotics and crystalloid fluids directly into the abdominal cavity. Abdominal exploration, collection of tissue samples, and local therapy is possible via standing laparoscopy in megavertebrates.


title: Flexibility as a Strategy in Nucleoside Antiviral Drug Design.

text: As far back as Melville Wolfrom's acyclic sugar synthesis in the 1960's, synthesis of flexible nucleoside analogues have been an area of interest. This concept, however, went against years of enzyme-substrate binding theory. Hence, acyclic methodology in antiviral drug design did not take off until the discovery and subsequent FDA approval of such analogues as Acyclovir and Tenofovir. More recently, the observation that flexible nucleosides could overcome drug resistance spawned a renewed interest in the field of nucleoside drug design. The next generation of flexible nucleosides shifted the focus from the sugar moiety to the nucleobase. With analogues such as Seley-Radtke "fleximers", and Herdewijn's C5 substituted 2'-deoxyuridines, the area of base flexibility has seen great expansion. More recently, the marriage of these methodologies with acyclic sugars has resulted in a series of acyclic flex-base nucleosides with a wide range of antiviral properties, including some of the first to exhibit anti-coronavirus activity. Various flexible nucleosides and their corresponding nucleobases will be compared in this review.


title: Notification of animal and human diseases: the global legal basis.

text: The successful control of a disease, and a possible epidemic, depends on rapid access to complete information on the disease situation. To ensure a timely response, diseases must be immediately notified in a transparent manner. The rapid exchange of information about animal diseases, including zoonoses, was the key objective in the establishment of the World Organisation for Animal Health (OIE) in 1924. For diseases concerning humans, a set of new rules dealing with the quick reporting of infectious diseases--the International Health Regulations--was adopted by Members of the World Health Organization (OMS) in 2005. The article explains these two systems of notification, which make information accessible to the public and allow decision-makers to better manage the risks related to the diseases concerned.


title: Intubation of SARS patients: infection and perspectives of healthcare workers.

text: PURPOSE The outbreak of severe acute respiratory syndrome (SARS) in 2003 presented major challenges to the safety of anesthesiologists and other healthcare workers (HCWs). This study determined the incidence of SARS transmission to HCWs who intubated patients and analyzed the concerns of HCWs regarding personal and patient safety. METHODS Healthcare workers who performed tracheal intubation in 10 Toronto hospitals were identified using the Ontario Public Health database. A questionnaire was used to collect information from the HCWs. To determine if the patterns of personal protection or concerns changed over time, data were analyzed according to whether the intubation occurred during SARS 1 (February 23 to April 21) or SARS 2 (April 22 to July 1). RESULTS Thirty-three HCWs who performed 39 intubations on 35 SARS patients were interviewed. Three of 23 HCWs (13%) acquired SARS during SARS 1 whereas none (0/10) acquired SARS during SARS 2. Personal protection increased from SARS 1 to SARS 2 and HCWs' concerns changed over time. During SARS 1, concerns focused on the need for personal protective equipment whereas during SARS 2, concerns focused on the need for strict training and patient care protocols. HCWs perceived that their experiences were ineffectively integrated into risk management protocols. CONCLUSIONS Protection guidelines failed to completely prevent the transmission of SARS to HCWs. Nine percent of the interviewed HCWs who intubated patients contracted SARS. A Risk Analysis Framework is presented to facilitate the rapid integration of HCWs' experiences into practice guidelines.


title: An investigation into the health status and diseases of wild dogs (Lycaon pictus) in the Kruger National Park.

text: Many factors have been cited as possible reasons for the decline in the number of wild dogs (Lycaon pictus), but few studies have provided supportive data. Between 1990 and 1993, the dynamics of 10 wild dog packs in the southern district of the Kruger National Park in South Africa were monitored. Casual observations of the causes of disease and mortality in the entire population were also recorded. During the same period, 46 wild dogs were immobilised, weighed, and subjected to physical examination as well as the collection of blood and faecal specimens. External parasites were collected and blood smears were prepared. Serum specimens were subjected to routine blood chemistry analyses, hormone and serological assays and formalinised blood specimens and faeces were subjected to screening for endoparasites. The study population varied from 75 in 1990 to 123 in 1993 with a survival rate of 29.9% for pups, 64.3% for yearlings and 69% for adults. Eighty per cent or more of the population were under 4 years of age. The cause of death in all wild dogs in the Kruger National Park could be established only in a small number of cases. Lions were responsible for the death of 20/62 wild dogs and disease caused the death of 6/62 wild dogs. All immobilised dogs were in a good physical condition, but 85% of dogs had one or more skin lesions. Potential life-threatening lesions (bitewounds inflicted by other dogs and lesions inflicted by a snare) occurred in 4 dogs. One male dog had only one testicle in the scortum. Twenty-six (93%) blood smears were positive for gametocytes of Hepatozoon sp., presumably H. canis, and in 2 dogs trophozoites of Babesia canis were seen. Eighty-six per cent of the specimens were positive for Dipetalonema reconditum. All dogs were infested with ticks and Haemaphysalis leachi, Amblyomma hebraeum, A. marmoreum, Boophilus decoloratus, Rhipicephalus simus, R. evertsi, R. appendiculatus and R. zambesiensis were identified. Ctenocephalides sp. and Echidnophaga larina were also identified. Taenia sp., Toxascaris canis and Ancylostoma caninum were present in faecal specimens. Antibody titres to adenovirus (26/31), B. canis (6/15), canine para-influenza virus (21/31), coronavirus (20/31), Coxiella burnetti (8/29), reovirus Type 3 (9/31), Rickettsia conori/africae (27/29), rotavirus (16/31) and Toxoplasma gondii (16/16) were found. The average serum urea concentration was higher (16.4 mmol/e) than that described for captive wild dogs, but other biochemical parameters were generally in agreement with values reported for captive wild dogs.(ABSTRACT TRUNCATED AT 400 WORDS)


title: COVID-19, cytokines and immunosuppression: what can we learn from severe acute respiratory syndrome?

text: A severe outbreak of coronavirus disease 2019 (COVID-19) emerged in China in December 2019, and spread so rapidly that more than 200,000 cases have so far been reported worldwide; on January 30, 2020, the WHO declared it the sixth public health emergency of international concern. The two previously reported coronavirus epidemics (severe acute respiratory syndrome [SARS] and Middle East respiratory syndrome [MERS]) share similar pathogenetic, epidemiological and clinical features as COVID-19. As little is currently known about SARS-CoV-2, it is likely that lessons learned from these major epidemics can be applied to the new pandemic, including the use of novel immunosuppressive drugs.


The 2014 Hajj and Umrah - current recommendations.


title: Identification of human aminopeptidase O, a novel metalloprotease with structural similarity to aminopeptidase B and leukotriene A4 hydrolase.

text: We have cloned and characterized a human brain cDNA encoding a new metalloprotease that has been called aminopeptidase O (AP-O). AP-O exhibits a series of structural features characteristic of aminopeptidases, including a conserved catalytic domain with a zinc-binding site (HEXXHX18E) that allows its classification in the M1 family of metallopeptidases or gluzincins. The structural complexity of AP-O is further increased by the presence of an additional C-terminal domain 170 residues long, which is predicted to have an ARM repeat fold originally identified in the Drosophila segment polarity gene product Armadillo. This ARM repeat domain is also present in aminopeptidase B, aminopeptidase B-like, and leukotriene A4 hydrolase and defines a novel subfamily of aminopeptidases that we have called ARM aminopeptidases. Northern blot analysis revealed that AP-O is mainly expressed in the pancreas, placenta, liver, testis, and heart. Human AP-O was produced in Escherichia coli, and the purified recombinant protein hydrolyzed synthetic substrates used for assaying aminopeptidase activity. This activity was abolished by general inhibitors of metalloproteases and specific inhibitors of aminopeptidases. Recombinant AP-O also cleaved angiotensin III to generate angiotensin IV, a bioactive peptide of the renin-angiotensin pathway with multiple actions on diverse tissues, including brain, testis, and heart. On the basis of these results we suggest that AP-O could play a role in the proteolytic processing of bioactive peptides in those tissues where it is expressed.


title: New York City's healthcare transportation during a disaster: a preparedness framework for a wicked problem.

text: During a disaster, victims with varied morbidities are located at incident sites, while healthcare facilities with varied healthcare resources are distributed elsewhere. Transportation serves an essential equilibrating role: it helps balance the patients' need for care with the supply of care. Studying the special case of New York City, this article sets out the healthcare transportation components as: (1) incident morbidity; (2) transportation assets; and (3) healthcare capacity. The relationship between these three components raises an assignment problem: the management of healthcare transportation within a dynamic and partly unpredictable incident-transportation-healthcare nexus, under urban disruption. While the routine dispatch problem can be tackled through better geographic allocation software and technical algorithms, the disaster assignment problem must be confronted through real-time, mutual adjustment between institutions. This article outlines institutional alternatives for managing the assignment problem and calls for further research on the merits of alternative institutional models.


title: Measures to Predict The Individual Variability of Corticospinal Responses Following Transcranial Direct Current Stimulation.

text: Individual responses to transcranial direct current stimulation (tDCS) are varied and therefore potentially limit its application. There is evidence that this variability is related to the contributions of Indirect waves (I-waves) recruited in the cortex. The latency of motor-evoked potentials (MEPs) can be measured through transcranial magnetic stimulation (TMS), allowing an individual's responsiveness to tDCS to be determined. However, this single-pulse method requires several different orientations of the TMS coil, potentially affecting its reliability. Instead, we propose a paired-pulse TMS paradigm targeting I-waves as an alternative method. This method uses one orientation that reduces inter- and intra-trial variability. It was hypothesized that the paired-pulse method would correlate more highly to tDCS responses than the single-pulse method. In a randomized, double blinded, cross-over design, 30 healthy participants completed two sessions, receiving 20 min of either anodal (2 mA) or sham tDCS. TMS was used to quantify Short interval intracortical facilitation (SICF) at Inter stimulus intervals (ISIs) of 1.5, 3.5 and 4.5 ms. Latency was determined in the posterior-anterior (PA), anterior-posterior (AP) and latero-medial (LM) coil orientations. The relationship between latency, SICF measures and the change in suprathreshold MEP amplitude size following tDCS were determined with Pearson's correlations. TMS measures, SICI and SICF were also used to determine responses to Anodal-tDCS (a-tDCS). Neither of the latency differences nor the SICF measures correlated to the change in MEP amplitude from pre-post tDCS (all P > 0.05). Overall, there was no significant response to tDCS in this cohort. This study highlights the need for testing the effects of various tDCS protocols on the different I-waves. Further research into SICF and whether it is a viable measure of I-wave facilitation is warranted.


title: A cross-sectional study on reference ranges of normal oral temperatures among students in Singapore.

text: INTRODUCTION During the 2003 Severe Acute Respiratory Syndrome (SARS) outbreak, all schools in Singapore implemented twice-daily temperature monitoring for students to curtail the spread of the disease. Students were not allowed to attend school if their temperature readings were >37.8 degrees C for students < or =12 years old, or > or =37.5 degrees C for students >12 years old. These values had been arbitrarily determined with professional inputs. The aim of this study is to determine the reference ranges of normal oral temperatures of students in Singapore and recommend the cut-off values for febrile patients. This may be used in another similar outbreak of an infectious disease with fever. MATERIALS AND METHODS Four co-ed primary schools and 4 co-ed secondary schools were selected for this study. Four thousand and two hundred primary 1 to secondary 3 students responded (96.8%) and participated in this cross-sectional study. The mean ages of the students in the lowest (primary 1) and highest educational levels (secondary 3) were 7.4 years old and 15.3 years old, respectively. Twelve oral temperature readings per student (i.e. measurements taken 4 times a day in 3 consecutive days) were collected. Forty-six thousand seven hundred and eighty-three (92.8%) out of 50,400 temperature readings were used for the analysis as missing data were excluded. A quantile regression model was applied to estimate reference ranges of normal oral temperatures for students with adjustment for potential confounding factors. RESULTS The age-specific reference ranges of normal oral temperature from this study for students < or =12 years old and >12 years old were 35.7 degrees C to 37.7 degrees C and 35.6 degrees C to 37.4 degrees C, respectively. Temperatures of 37.8 degrees C and 37.5 degrees C are therefore recommended as the oral temperature cut-offs for those < or =12 years old and >12 years old, respectively. CONCLUSION This study has provided empirical data on normal oral temperature cut-offs which could be used during temperature screening in schools.


title: Severe respiratory illness caused by a novel coronavirus, in a patient transferred to the United Kingdom from the Middle East, September 2012.

text: Coronaviruses have the potential to cause severe transmissible human disease, as demonstrated by the severe acute respiratory syndrome (SARS) outbreak of 2003. We describe here the clinical and virological features of a novel coronavirus infection causing severe respiratory illness in a patient transferred to London, United Kingdom, from the Gulf region of the Middle East.


title: Programmed translational frameshifting is likely required for expressions of genes encoding putative nuclear protein kinases of the ciliate Euplotes octocarinatus.

text: Three macronuclear genes encoding putative nuclear protein kinases of the ciliate Euplotes octocarinatus syngen 1 were isolated and sequenced. All three deduced gene products share significant properties with a group of recently identified nuclear serine/threonine protein kinases named Ndr. The three predicted proteins contain the twelve conserved catalytic subdomains of protein kinases and 22 near universally-conserved amino acids residues that are characteristic of serine/threonine protein kinases. In addition, there is an approximately 30 amino acid-peptide insertion between subdomains VII and VIII that contains a potential nuclear localization signal. Sequence analysis suggests that expression of the Eondr2 gene requires a + 1 programmed translational frameshift for its translation. Comparison of the deduced EoNdr2 with other known Ndr protein kinases implies that a + 1 ribosomal frameshift occurs at the motif AAATAA.


Does screening keep Ebola out of USA?


Current understanding of vaccination site-associated sarcomas in the cat.


title: Ecological origins of novel human pathogens.

text: A systematic literature survey suggests that there are 1399 species of human pathogen. Of these, 87 were first reported in humans in the years since 1980. The new species are disproportionately viruses, have a global distribution, and are mostly associated with animal reservoirs. Their emergence is often driven by ecological changes, especially with how human populations interact with animal reservoirs. Here, we review the process of pathogen emergence over both ecological and evolutionary time scales by reference to the "pathogen pyramid." We also consider the public health implications of the continuing emergence of new pathogens, focusing on the importance of international surveillance.


title: Multiple receptor-dependent steps determine the species specificity of HCV-229E infection.

text: Human coronavirus (HCV)-229E causes disease only in humans and grows in human cells and in cells of other species that express recombinant human aminopeptidase N (hAPN), the receptor for HCV-229E. We compared the species specificity of HCV-229E infection with the species specificity of virus binding using immunofluorescence, assay of virus yields, fluorescence activated cell sorting and a monoclonal antibody directed against hAPN that blocks infection. We found that HCV-229E binds to intestinal brush border membranes (BBM) and to membranes of cell lines from cats, dogs, pigs, and humans, however the virus only infects two of these species. HCV-229E will not bind to BBM or to membranes from cell lines derived from hamster or mice. Animal coronaviruses related to HCV-229E, including FIPV, CCV, and TGEV bind to cell membranes from cats, dogs, cows, pigs and humans (but not mice), while each virus infects cells from only a subset of these species. Infectious genomic HCV-229E RNA, can infect cells of all of these species. These data suggest that the species-specificity of infection for this serogroup of coronaviruses is determined at the levels of virus binding and penetration. Since binding of viral spike glycoprotein to cellular receptors is not the only limiting factor, we suggest that one or more steps associated with virus penetration may determine the species specificity of infection with the HCV-229E serogroup of coronaviruses.


title: Astrovirus induces diarrhea in the absence of inflammation and cell death.

text: Astroviruses are a leading cause of infantile viral gastroenteritis worldwide. Very little is known about the mechanisms of astrovirus-induced diarrhea. One reason for this is the lack of a small-animal model. Recently, we isolated a novel strain of astrovirus (TAstV-2) from turkeys with the emerging infectious disease poult enteritis mortality syndrome. In the present studies, we demonstrate that TAstV-2 causes growth depression, decreased thymus size, and enteric infection in infected turkeys. Infectious TAstV-2 can be recovered from multiple tissues, including the blood, suggesting that there is a viremic stage during infection. In spite of the severe diarrhea, histopathologic changes in the intestine were mild and there was a surprising lack of inflammation. This may be due to the increased activation of the potent immunosuppressive cytokine transforming growth factor beta during astrovirus infection. These studies suggest that the turkey will be a useful small-animal model with which to study astrovirus pathogenesis and immunity.


Organizational aspects of intensive care unit resource allocation in a primary respiratory intensive care center specialized for the treatment of SARS-COV-2 patients.


title: Broad cellular immunity with robust memory responses to simian immunodeficiency virus following serial vaccination with adenovirus 5- and 35-based vectors.

text: Adenovirus serotype 35 (Ad35) is a promising vaccine platform for human immunodeficiency virus (HIV) infection and emerging infectious diseases as it is uncommon in humans worldwide and is distinct from Ad5, the major vaccine serotype for which many individuals have pre-existing immunity. The immunogenicity of a first-generation, replication-competent Ad35-based vaccine was tested in the simian immunodeficiency virus (SIV) rhesus macaque model by evaluating its capacity to boost immunity generated by Ad5-based vectors. A series of four immunizations with replication-defective Ad5 vectors expressing SIVmac239 gag induced high-frequency responses mediated by both CD8(+) and CD4(+) T cells directed against several epitopes. Ad5-specific neutralizing antibody responses that did not neutralize Ad35 were rapidly induced but waned over time. Subsequent immunization with Ad5-based vectors was minimally effective, whereas immunization with Ad35-based vectors generated a strong increase in the frequency of Gag-specific T cells with specificities that were unchanged. While this boosting response was relatively transient, challenge with the distinct pathogenic isolate SIV/DeltaB670 generated robust and selective recall responses to Gag with similar specificities as induced by vaccination that were elevated for 25 weeks relative to controls. Vaccination had measurable albeit minor effects on virus load. Unexpectedly, regional hypervariability within the Gag sequence of SIV/DeltaB670 was associated with mutation of the conserved CD8(+) T-cell epitope CM9 without concurrent flanking mutations and in the absence of immune pressure. These findings support the further development of Ad35 as a vaccine vector, and promote vaccine regimens that utilize serial administration of heterologous adenoviruses.