title: T cell responses to whole SARS coronavirus in humans.

text: Effective vaccines should confer long-term protection against future outbreaks of severe acute respiratory syndrome (SARS) caused by a novel zoonotic coronavirus (SARS-CoV) with unknown animal reservoirs. We conducted a cohort study examining multiple parameters of immune responses to SARS-CoV infection, aiming to identify the immune correlates of protection. We used a matrix of overlapping peptides spanning whole SARS-CoV proteome to determine T cell responses from 128 SARS convalescent samples by ex vivo IFN-gamma ELISPOT assays. Approximately 50% of convalescent SARS patients were positive for T cell responses, and 90% possessed strongly neutralizing Abs. Fifty-five novel T cell epitopes were identified, with spike protein dominating total T cell responses. CD8(+) T cell responses were more frequent and of a greater magnitude than CD4(+) T cell responses (p < 0.001). Polychromatic cytometry analysis indicated that the virus-specific T cells from the severe group tended to be a central memory phenotype (CD27(+)/CD45RO(+)) with a significantly higher frequency of polyfunctional CD4(+) T cells producing IFN-gamma, TNF-alpha, and IL-2, and CD8(+) T cells producing IFN-gamma, TNF-alpha, and CD107a (degranulation), as compared with the mild-moderate group. Strong T cell responses correlated significantly (p < 0.05) with higher neutralizing Ab. The serum cytokine profile during acute infection indicated a significant elevation of innate immune responses. Increased Th2 cytokines were observed in patients with fatal infection. Our study provides a roadmap for the immunogenicity of SARS-CoV and types of immune responses that may be responsible for the virus clearance, and should serve as a benchmark for SARS-CoV vaccine design and evaluation.


title: The causes of glaucoma in cats.

text: The cause of glaucoma in 131 enucleated eyes from 128 cats was determined in a retrospective histologic study. Obliteration of the ciliary cleft by diffuse iridal melanoma (38 eyes), or other neoplasms (14 eyes), or by the presence of idiopathic lymphocytic-plasmacytic anterior uveitis (53 eyes) were the most frequent lesions likely to explain the development of glaucoma. Secondary changes of inner retinal atrophy, optic disc cupping, scleral thinning with megaglobus, and atrophy of ciliary processes were similar to those described in dogs and human beings with chronic glaucoma. In light of the duration and severity of the glaucoma, the degree of inner retinal atrophy was often less than expected. Diffuse corneal edema and breaks in Descemet's membrane, changes typical of glaucoma in other species, were rarely detected. Eyes with chronic uveitis and glaucoma had collapsed ciliary clefts, iridoscleral adhesions, and posterior displacement of the iris. We were unable to determine whether these changes were consequences of the uveitis and thus responsible for the development of glaucoma, or if they were merely the result of the chronic glaucoma itself.


New European research project to respond to unexpected epidemic threats such as Escherichia coli.


title: Development of vaccines against common colds.

text: Respiratory tract viruses are particularly significant causes of illness and death in children and in the elderly. Vaccines offer the possibility of decreasing the severity and complications of viral respiratory disease, but development has been delayed by numerous factors. First, there are more than 200 serologically distinct RNA and DNA virus species and strains which cause an essentially similar spectrum of disease. Some re-infect at high efficiency despite little antigenic variation, while others exhibit extensive coat protein variability. Vaccine candidates show variable efficacy in partially immune adults, the immunocompromised and the elderly, and may be ineffective or pathogenic in neonates or in the presence of maternal antibodies. However, effective childhood vaccines are essential to prevent severe disease due to respiratory syncytial virus (RSV) and parainfluenza and to reduce virus transmission to adults. A number of promising vaccines are in clinical trial, and it is likely that vaccines against RSV and parainfluenza will be licensed within the next 5-10 years. Mucosal delivery and the use of novel adjuvants offers the prospect of better vaccines against influenza. The ultimate goal is to develop multivalent mucosal vaccines offering protection against a spectrum of respiratory infections.


title: [Laboratory on a microfluidic chip].

text: The recent achievements of microfluidic chip and its applications, based on the works mainly carried out in the authors' lab are reviewed. The chip fabrication capabilities have been extended into design and fabricate chips with higher degree of complexity in different materials, such as quartz, glass, polymethyl methacrylate (PMMA), and polydimethyl siloxane (PDMS). A set of methods for surface modification of micro-channels on such materials have been developed, which results in better reproducibility and higher efficiency in protein and peptide analysis. The use of novel materials for chip fabrication is also under investigation. A series of microfluidic workstations with integrated chip manipulation as well as laser induced fluorescence (LIF), ultraviolet (UV), electrochemical and chemiluminescence detection modules have been developed to attain the abilities of complex microfluidic control and data acquisition schemes. A single cell/single molecule imagining system was built up for dynamic analysis of molecular or cellular events too. Based on the work mentioned above, different functional units, such as membrane, monolithic, isotachophoresis (ITP) etc were set up and integrated. Glycoform separation of turkey ovalbumin in a lectin monolithic column and an electrophoresis channel was performed on an integrated microchip. And a novel technique has been developed that allows for the coupling of ITP and non-gel sieving electrophoresis for protein analysis in a single microchip and resulting in - 50 fold increase of the sensitivity in comparison with the use of gel electrophoresis only. A single molecule detection (SMD) based technique was developed for simultaneously measuring both bulk flow and near-wall flow velocity in the microchannels. And more recently, an SMD based technology was developed for observing molecular interactions at single molecule level. An ultra-rapid microchip electrophoresis method was established for simultaneous determination intracellular reactive oxygen species (ROS) and reduced glutathione (GSH) related to apoptosis and oxidative stress. In an effort to develop a novel microfluidic based drug screening platform, systematic studies on the interaction between granulocyte colony-stimulating factor (G-CSF) and sulfated oligosaccharides were carried out at both molecular and cellular levels. Doxorubicin induced apoptosis of human hepatocellular carcinoma (HepG2) was studied using the integrated microfluidic device with concentration generator. In the application phase, severe acute respiratory syndrome (SARS) diagnosis based on reverse transcription-polymerase chain reaction (RT-PCR) and microfluidic chip electrophoresis (MCE) with 18 cases, methylation analysis of the P16 gene in 159 samples of patients and references for cancer diagnosis and polymorphism analysis of angiotenigen gene in 226 patients and references with essential hypertension are described. Forty-three up to date references are


title: Risk-stratified seroprevalence of severe acute respiratory syndrome coronavirus among children in Hong Kong.

text: BACKGROUND Severe acute respiratory syndrome was relatively mild in children, and the incidence was significantly lower when compared with adults. Although previous seroepidemiological studies demonstrated that asymptomatic infection was uncommon among health care workers and adult contacts of patients with severe acute respiratory syndrome, it is unclear whether this would extend to the pediatric population. OBJECTIVE The purpose of this study was to determine the seroprevalence of severe acute respiratory syndrome coronavirus among asymptomatic children living near Amoy Gardens (site of largest community outbreak of severe acute respiratory syndrome in Hong Kong) compared with a low-risk region where no community transmission occurred. METHODS The study was conducted from September to October 2003. Target subjects living in the defined high-risk and low-risk areas were approached through the schools within the respective localities. We recruited 353 and 361 children, respectively, from the high-risk and low-risk areas and collected 3 to 5 mL of blood for severe acute respiratory syndrome coronavirus IgG antibody testing by immunofluorescence antibody assay and confirmation by neutralization test. Parents of all of the subjects who joined the study were contacted by telephone, and a standardized questionnaire was administered by a research nurse to collect information including sociodemographic data, history of severe acute respiratory syndrome coronavirus infection in the subjects and members of the household, history of contact with known cases of severe acute respiratory syndrome, presence of severe acute respiratory syndrome-like symptoms since onset of the severe acute respiratory syndrome epidemic, travel history of the child and his/her relatives within the 15 days before any such symptom onset, use of health service as a result of such symptoms, and whether there were deaths of relatives as a result of severe acute respiratory syndrome. RESULTS Two (0.57%) of 353 asymptomatic children from the high-risk area were tested positive for severe acute respiratory syndrome coronavirus antibody compared with 0 of 361 in the low-risk region. None of the 14 children who lived in the high-risk area and had known contacts with severe acute respiratory syndrome patients were seropositive. CONCLUSIONS As in adults, subclinical severe acute respiratory syndrome coronavirus infection was rare in children in the 2003 epidemic. The very low seroprevalence implies little or no population herd immunity to protect against future resurgence of severe acute respiratory syndrome.


Recombination in RNA viruses and in virus-resistant transgenic plants.


title: Serositis: comparative analysis of histological findings and pathogenetic mechanisms in nonbacterial serosal inflammation.

text: Peritonitis is the established term for infective inflammation of the peritoneum, while serositis generally refers to nonorganismal inflammation in any serous cavity, including the peritoneum. In continuous ambulatory peritoneal dialysis (CAPD) literature, however, culture-negative peritoneal inflammation is referred to as "sterile" or "chemical" peritonitis. These terms not only imply unwarranted etiologic assumptions, but may also deflect attention from the existence of medical conditions to which the peritoneum is subject. This is evident in CAPD literature where there is little recognition that the peritoneum, as a member of the serosa and a secretor of lamellar bodies, is prey to a wide range of disorders. Thus before, during, and after CAPD, the membrane is liable to fall victim to disease states unconnected with the process of dialysis. Significant peritoneal pathology occurs as part of a pan-serositis, which may be metabolic (uremia, cholesterolosis), autoimmune (systemic lupus erythematosus, rheumatoid disease, acute rheumatism, endocrinopathies), genetic (recurrent hereditary polyserositis), allergic (eosinophilic serositis), and granulomatous in nature. This paper presents a comparative analysis of histopathological presentation and pathogenetic mechanisms involved in all forms of peritoneal serositis. It incorporates recent advances in molecular biology of the membrane into a holistic reappraisal of peritoneal pathology, revealing hitherto unrecognized homologies in peritoneal reaction to diverse disorders.


title: Next revolution in the molecular theranostics of infectious diseases: microfabricated systems for personalized medicine.

text: The molecular diagnosis of infectious diseases is currently going through a revolution sustained by the regulatory approval of amplification tests that have been shown to be equivalent or superior to existing gold standard methods. The recent approval of a microarray system for the pharmacogenomic profiling of cytochrome P450-mediated drug metabolism is paving the way to novel, rapid, sensitive, robust and economical microfabricated systems for point-of-care diagnostics, which are utilized closer and closer to the patient's bedside. These systems will enable the multiparametric genetic evaluation of several medical conditions, including infectious diseases. This forecoming revolution will position molecular theranostics in a broader integrated view of personalized medicine, which exploits genetic information from microbes and human hosts to optimize patient management and disease treatment.


title: [Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein].

text: OBJECTIVE To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain. METHODS According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA. RESULTS Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained. CONCLUSIONS The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.


Avian infectious bronchitis.


title: Complete port-accessed lobectomy by the muscle-sparing method.

text: We report the case of a 53-year-old woman who underwent complete port-accessed middle lobectomy by a new technique that preserves all muscles, including the extracostal and intercostal muscles. The operation was performed by using only thoracovideoscopy, and the resected lobe was withdrawn in a pouch through a subxiphoid incision through the substernal route. This complete port-accessed lobectomy is a new technique and is thought to be less invasive than video-assisted lobectomy with minithoracotomy.


title: Remodeling the endoplasmic reticulum by poliovirus infection and by individual viral proteins: an autophagy-like origin for virus-induced vesicles.

text: All positive-strand RNA viruses of eukaryotes studied assemble RNA replication complexes on the surfaces of cytoplasmic membranes. Infection of mammalian cells with poliovirus and other picornaviruses results in the accumulation of dramatically rearranged and vesiculated membranes. Poliovirus-induced membranes did not cofractionate with endoplasmic reticulum (ER), lysosomes, mitochondria, or the majority of Golgi-derived or endosomal membranes in buoyant density gradients, although changes in ionic strength affected ER and virus-induced vesicles, but not other cellular organelles, similarly. When expressed in isolation, two viral proteins of the poliovirus RNA replication complex, 3A and 2C, cofractionated with ER membranes. However, in cells that expressed 2BC, a proteolytic precursor of the 2B and 2C proteins, membranes identical in buoyant density to those observed during poliovirus infection were formed. When coexpressed with 2BC, viral protein 3A was quantitatively incorporated into these fractions, and the membranes formed were ultrastructurally similar to those in poliovirus-infected cells. These data argue that poliovirus-induced vesicles derive from the ER by the action of viral proteins 2BC and 3A by a mechanism that excludes resident host proteins. The double-membraned morphology, cytosolic content, and apparent ER origin of poliovirus-induced membranes are all consistent with an autophagic origin for these membranes.


title: Outbreak of pandemic influenza A (H1N1-2009) in Singapore, May to September 2009.

text: INTRODUCTION The fi rst case of pandemic influenza A(H1N1) was detected in Singapore on 26 May 2009, 1 month after the fi rst cases of novel influenza A(H1N1) was reported in California and Texas in the United States. The World Health Organization declared the fi rst influenza pandemic of the 21st century on 11 June 2009. MATERIALS AND METHODS Confirmed cases notified to the Ministry of Health between 27 May and 9 July 2009 were analysed. Various indicators of influenza activity were monitored throughout the study period. Estimates of the number of cases of H1N1-2009 were made using the number of polyclinic attendances for acute respiratory infection and influenza-like illness and the weekly prevalence of H1N1-2009. RESULTS Cases in Singapore affected mainly young adults, youths and children. By the end of September 2009, it was estimated that at least 270,000 persons had been infected by pandemic influenza A (H1N1) in Singapore. The peak number of cases occurred during E-week 30 (26 July-1 August) when an estimated 45,000 cases were seen in polyclinics and GP clinics. The hospitalisation, severe illness and mortality rates were estimated at 6 per 1000 cases, 0.3 per 1000 cases and 6.7 per 100,000 cases, respectively. The most common risk factors among hospitalised adult cases were asthma and diabetes. For hospitalised children, the most common risk factors were being under 5 years of age and asthma. The most common risk factors among persons with severe illness were diabetes in adults and epilepsy and being under 5 years of age in children. About half of cases with severe illness required mechanical ventilation. In addition, one-fifth of cases with severe illness had acute respiratory distress syndrome. CONCLUSIONS The fi rst wave of the influenza pandemic lasted about 10 weeks. Morbidity and mortality resulting from pandemic influenza were low.


title: Spread of a neurotropic coronavirus to spinal cord white matter via neurons and astrocytes.

text: Mouse hepatitis virus strain JHM (MHV-JHM) causes a chronic encephalomyelitis in susceptible mice, with histological evidence of demyelination in the spinal cord. After intranasal inoculation, virus spreads retrogradely to several brain structures along neuroanatomic projections to the main olfactory bulb. In the absence of experimental intervention, mice become moribund before the spinal cord is infected. In this study, infusions of anti-MHV neutralizing monoclonal antibodies were administered to protect mice from the MHV-JHM-induced acute encephalitis and to allow survival until virus spread to the spinal cord. Under these conditions, virus was observed to enter specific layers (primarily laminae V to VII) in the gray matter of the upper spinal cord, consistent with transneuronal spread. While the brain structures which are the sources for virus spread to the spinal cord cannot be determined with certainty, the ventral reticular nucleus is likely to be important since it is consistently and extensively labeled in all mice and receives projections from subsequently infected areas of the spinal cord. After initial entry into the gray matter, virus rapidly spread to the white matter of the spinal cord. During the early stages of this process, extensive infection of astrocytes was noted, suggesting that cell-to-cell spread via these glial cells is an important part of this process. Reports from other laboratories using cultured cells strongly suggested that astrocytes serve as important regulators of oligodendrocyte function and, by extrapolation, have a major role in vivo in the processes of both demyelination and remyelination. Thus, our results not only outline the probable pathway used by MHV-JHM to infect the white matter of the spinal cord but also, with the assumption that infection of astrocytes leads to subsequent dysfunction, raise the possibility that infection of these cells contributes to the demyelinating process.


title: Disassociation between the in vitro and in vivo effects of nitric oxide on a neurotropic murine coronavirus.

text: Intranasal inoculation of the neuroattenuated OBLV60 strain of mouse hepatitis virus results in infection of mitral neurons in the olfactory bulb, followed by spread along olfactory and limbic pathways to the brain. Immunocompetent BALB/c mice were able to clear virus by 11 days postinfection (p.i.). Gamma interferon (IFN-gamma) may play a role in clearance of OBLV60 from infected immunocompetent BALB/c mice through a nonlytic mechanism. Among the variety of immunomodulatory activities of IFN-gamma is the induction of expression of inducible nitric oxide synthase (iNOS), an enzyme responsible for the production of nitric oxide (NO). Studies were undertaken to investigate the role of IFN-gamma and NO in host defense and clearance of OBLV60 from the central nervous system (CNS). Exposure of OBLV60-infected OBL21a cells, a mouse neuronal cell line, to the NO-generating compound S-nitroso-L-acetyl penicillamine resulted in a significant decrease in viral replication, indicating that NO interfered with viral replication. Furthermore, infection of IFN-gamma knockout (GKO) mice and athymic nude mice with OBLV60 resulted in low-level expression of iNOS mRNA and protein in the brains compared to that of OBLV60-infected BALB/c mice. Nude mice were unable to clear virus and eventually died between days 11 and 14 p.i. (B. D. Pearce, M. V. Hobbs, T. S. McGraw, and M. J. Buchmeier, J. Virol. 68:5483-5495, 1994); however, GKO mice survived infection and cleared virus by day 18 p.i. These data suggest that IFN-gamma production in the olfactory bulb contributed to but may not be essential for clearance of OBLV60 from the brain. In addition, treatment of OBLV60-infected BALB/c mice with aminoguanidine, a selective inhibitor of iNOS activity, did not result in any increase in mortality, and the mice cleared the virus by 11 days p.i. These data suggest that although NO was able to block replication of virus in vitro, expression of iNOS with NO release in vivo did not appear to be the determinant factor in clearance of OBLV60 from CNS neurons.


title: Evaluation of the novel respiratory virus surveillance program: Pediatric Early Warning Sentinel Surveillance (PEWSS).

text: OBJECTIVES Infections caused by respiratory viruses are associated with recurrent epidemics and widespread morbidity and mortality. Routine surveillance of these pathogens is necessary to determine virus activity, monitor for changes in circulating strains, and plan for public health preparedness. The Southern Nevada Health District in Las Vegas, Nevada, recruited five pediatric medical practices to serve as sentinel sites for the Pediatric Early Warning Sentinel Surveillance (PEWSS) program. METHODS Sentinel staff collected specimens throughout the year from ill children who met the influenza-like illness case definition and submitted specimens to the Southern Nevada Public Health Laboratory for molecular testing for influenza and six non-influenza viruses. RESULTS Laboratory results were analyzed and reported to the medical and general communities in weekly bulletins year-round. PEWSS data were also used to establish viral respiratory seasonal baselines and in influenza vaccination campaigns. The surveillance program was evaluated using the Centers for Disease Control and Prevention's (CDC's) Updated Guidelines for Evaluating Public Health Surveillance Systems. PEWSS met three of six program usefulness criteria and seven of nine surveillance system attributes, which exceeded the CDC Guidelines evaluation criteria for a useful and complete public health surveillance program. CONCLUSION We found that PEWSS is a useful and complete public health surveillance system that is simple, flexible, accessible, and stable.


title: Natural West Nile virus infection in a captive juvenile Arctic wolf (Canis lupus).

text: A case of West Nile virus (WNV) infection in a captive 4-month-old Arctic wolf (Canis lupus) is described. The animal had vomiting, anorexia, and ataxia before death. Histopathology revealed multifocal severe renal lymphoplasmacytic vasculitis, mostly affecting small arterioles, with fibrinoid degeneration of some vessel walls. Many small foci of gliosis were detected in the cerebral cortex. West Nile virus was demonstrated in the kidneys and cerebrum by immunohistochemistry and polymerase chain reaction. The described renal changes represent a novel pathological finding of WNV infection.


title: Adult respiratory distress syndrome.

text: ARDS or acute respiratory distress syndrome continues to be a considerable critical care challenge. Mortality has not decreased significantly over the last more than 30 years. This article presents an overview of origin, evaluation, and treatment of ARDS. Recent findings relative to onset and precipitators of ARDS have led to changes in evaluation and treatment plans. Clinical and radiologic descriptors in assessment of the patient with ARDS are discussed. Ventilatory modes and nursing interventions to optimize patient outcomes are identified. The challenges of outcomes issues presented offer opportunities for further study.


Use of humanised mice to study antiviral activity of human γδ-T cells against influenza A viruses.


title: [A cell line particularly susceptible to bovine enteric Coronavirus replication: HRT 18 cells].

text: Bovine enteric Corona virus (BEC) multiplies spontaneously with very high efficiency in HRT 18 cells; this line established from human rectum adenocarcinoma presents some properties of the differentiated brush border cell of the intestinal villi. Titers obtained are very high (5 x 10(7) TCID 50/ml virus strain F15). The virus produced had the characteristic of BEC: shape, spikes, density 1.19, hemadsorption and hemagglutination of rat erythrocytes. Human and dog enteritic corona viruses replicate also in HRT 18 cells.


title: Immunopathogenesis and immunotherapy of multiple sclerosis.

text: Multiple sclerosis (MS) is a chronic disease of the CNS that is characterized by inflammation, demyelination and axonal injury. Although the etiology of MS is still unknown, many findings point toward a central role for the immune system in the pathogenesis of the disease. This hypothesis is strongly supported by the beneficial effects of immunomodulatory and immunosuppressive therapy on disease activity. Over the past few years, substantial progress has been made in deciphering the immune response in MS. Although animal models have advanced our knowledge of basic mechanisms of immune responses in the CNS, recent studies have also highlighted the differences between MS and its animal equivalent, experimental autoimmune encephalomyelitis. New immunotherapeutic agents have been developed and evaluated in clinical trials. Here, we review current knowledge of the immunopathogenesis of MS and corresponding animal models of disease, and discuss new immunointerventional treatment strategies based on changing pathogenetic concepts.


Racial discrimination and mental illness.


title: Molecular characterization and typing of chicken and turkey astroviruses circulating in the United States: implications for diagnostics.

text: Avian astroviruses were detected by reverse transcriptase and polymerase chain reaction in intestinal contents collected from commercial chickens and turkeys from throughout the United States from 2003 through 2005. Astroviruses were detected in birds from both healthy and poorly performing flocks with or without enteric disease. Phylogenetic analysis was performed with sequence data from the polymerase (ORF-1b) genes of 41 turkey-origin astroviruses and 23 chicken-origin astroviruses. All currently available avian astrovirus sequence data and selected mammalian astrovirus sequence data were included in the analysis. Four groups of avian astroviruses were observed by phylogenetic analysis: turkey astrovirus type 1 (TAstV-1)-like viruses, turkey astrovirus type 2 (TAstV-2)-like viruses, both detected in turkeys; avian nephritis virus (ANV)-like viruses, detected in both chickens and turkeys; and a novel group of chicken-origin astroviruses (CAstV). Among these four groups, amino acid identity was between 50.1% and 73.8%, and was a maximum of 49.4% for all avian isolates when compared with the mammalian astroviruses. There were multiple phylogenetic subgroups within the TAstV-2, ANV, and CAstV groups based on 9% nucleotide sequence divergence. Phylogenetic analysis revealed no clear assortment by geographic region or isolation date. Furthermore, no correlation was observed between the detection of a particular astrovirus and the presence of enteric disease or poor performance. Based on these data, a revision of the present taxonomic classification for avian astroviruses within the genus Avastrovirus is warranted.


title: Nucleotide sequence of the gene encoding the spike glycoprotein of human coronavirus HCV 229E.

text: The gene encoding the spike glycoprotein of the human coronavirus HCV 229E has been cloned and sequenced. This analysis predicts an S polypeptide of 1173 amino acids with an Mr of 128,600. The polypeptide has 30 potential N-glycosylation sites. A number of structural features typical of coronavirus S proteins can be recognized, including a signal sequence, a membrane anchor, heptad repeat structures and a carboxy-terminal cysteine cluster. A detailed, computer-aided comparison with the S proteins of infectious bronchitis virus, feline infectious peritonitis virus, transmissible gastroenteritis virus and murine hepatitis virus, strain JHM is presented. We have also done a Northern blot analysis of viral RNAs in HCV 229E-infected cells using synthetic oligonucleotides. On the basis of this analysis, and by analogy to the replication strategy of other coronaviruses, we are able to propose a model for the organization and expression of the HCV 229E genome.


title: [What is the origin of SARS-CoV-2?]

text: Every time a pandemic occurs, dozens of theories emerge to attribute the origin of the event to different facts. The COVID-19 pandemic that has hit virtually all the globe has been no exception. What is known so far about the origin of the virus that causes COVID 19? The first investigations on the origin of this disease have determined that it is a new type of virus, the origin of which is most likely zoonotic.


title: Emergence of a new genotype of avian infectious bronchitis virus in Brazil.

text: Infectious bronchitis virus (IBV) is the agent of a highly contagious disease that affects domestic fowl (Gallus gallus). Recent reports showed a high prevalence of one main IBV genotype (Brazil or BR-I) with low genetic diversity in commercial poultry flocks from Brazil. This research analyzed IBV positive poultry flocks from different rearing regions to verify the S1 gene variability and geographic distribution of variant IBV strains in recent years (2010 and 2011). Samples of IBV-positive flocks were obtained from 60 different farms. Forty-nine partial S1 gene sequences were determined and aligned for phylogenetic and amino acid similarity analyses. Eleven samples (22.4%) were similar to Massachusetts vaccine strains (Mass genotype) and 34 samples (69.4%) to the previously characterized Brazilian BR-I genotype. Interestingly, the remaining four samples (8.2%) clustered into a new IBV variant genotype (Brazil-II or BR-II), divergent from the BR-I. A unique nucleotide sequence insertion coding for five amino acid residues was observed in all the Brazilian variant viruses (BR-I and BR-II genotypes). These results show a higher genetic diversity in Brazilian IBV variants than previously described.


title: Immunotoxicity of monoclonal antibodies.

text: Monoclonal antibodies (mAbs) are large molecules intended to bind to specific targets often expressed on the immune system, and to treat various immunopathological conditions. Therefore, mAbs can be considered to have a high potential for immunotoxicity, which is reflected in the clinical experience accumulated on mAbs-induced adverse effects related to immunosuppression, immunostimulation and hypersensitivity (immunogenicity). So far, non clinical immunotoxicity studies have been inadequate to address all safety issues in relation to the possible immunotoxicity of mAbs, because they are fraught with limitations and pitfalls primarily related to the lack of relevant animal species. In addition, clinical studies rarely include validated end-points dedicated to the prediction of immunotoxicity. With the ongoing development of mAbs as novel therapeutic strategies for a wide variety of diseases, efforts should be paid to improve our understanding of mAbs-induced immunotoxic effects and design dedicated strategies to assess their immunological safety, both non clinically and clinically.


title: The 5'-end sequence of the genome of Aichi virus, a picornavirus, contains an element critical for viral RNA encapsidation.

text: Picornavirus positive-strand RNAs are selectively encapsidated despite the coexistence of viral negative-strand RNAs and cellular RNAs in infected cells. However, the precise mechanism of the RNA encapsidation process in picornaviruses remains unclear. Here we report the first identification of an RNA element critical for encapsidation in picornaviruses. The 5' end of the genome of Aichi virus, a member of the family Picornaviridae, folds into three stem-loop structures (SL-A, SL-B, and SL-C, from the most 5' end). In the previous study, we constructed a mutant, termed mut6, by exchanging the seven-nucleotide stretches of the middle part of the stem in SL-A with each other to maintain the base pairings of the stem. mut6 exhibited efficient RNA replication and translation but formed no plaques. The present study showed that in cells transfected with mut6 RNA, empty capsids were accumulated, but few virions containing RNA were formed. This means that mut6 has a severe defect in RNA encapsidation. Site-directed mutational analysis indicated that as the mutated region was narrowed, the encapsidation was improved. As a result, the mutation of the 7 bp of the middle part of the stem in SL-A was required for abolishing the plaque-forming ability. Thus, the 5'-end sequence of the Aichi virus genome was shown to play an important role in encapsidation.


title: Herpes simplex virus, meningitis and encephalitis in neonates.

text: The consequences of neonatal herpes simplex virus (HSV) infection can be severe. Disease can be localized to skin, eye and mouth (SEM disease), involve the central nervous system (CNS) or manifest as disseminated infection involving multiple organs. Most surviving infants in the latter two categories have neurological sequelae, and the mortality rate in the absence of therapy is very high (80%) for babies in the latter category. The International Herpes Management Forum (IHMF) has produced guidelines on the diagnosis, prevention and effective management of neonatal herpes. Neonatal herpes may occur in the absence of skin lesions, so if the infection is suspected, swabs of the oropharynx, conjunctiva, rectum, skin lesions, mucosal lesions and urine should be promptly taken and submitted for virus culture. Cerebrospinal fluid (CSF) should be submitted for polymerase chain reaction (PCR) detection of HSV DNA. Evidence for disseminated or CNS infection should be sought using liver function tests, complete blood cell count, CSF analysis and chest X-ray, if respiratory abnormalities are present. Neonates with suspected HSV infection should be treated with intravenous aciclovir (20 mg/kg) every 8 h for 21 days. If disease is localized to the SEM, treatment should be limited to 14 days. The neutrophil count for children receiving intravenous aciclovir should be monitored. If the absolute neutrophil count falls below 500/mm3, decreasing the aciclovir dose or administering granulocyte colony stimulating factor (GCSF) should be considered. At the end of therapy in CNS and disseminated disease, PCR assessment of CSF should be used and treatment continued if the child remains PCR positive at this site.


title: Morphologic changes in the nasal cavity associated with sialodacryoadenitis virus infection in the Wistar rat.

text: A sequential study of lesions of the nasal cavity associated with sialodacryoadenitis virus (SDAV) infection was made in the laboratory rat. Wistar rats were intranasally inoculated with approximately 10(3) TCID50 of the coronavirus SDAV. Transverse sections of four regions of the nasal cavity from inoculated and control animals were examined by light microscopy and immunohistochemistry at 2, 4, 6, 8, 10, and 14 days postinoculation (PI). Lesions were observed in the following regions of the upper respiratory tract: respiratory epithelium, transitional epithelium, olfactory epithelium, nasolacrimal duct, vomeronasal organ, and the submucosal glands of the nasal passages. In general, in structures lined by ciliated epithelial cells, there was focal to segmental necrosis with exfoliation of affected cells and polymorphonuclear cell infiltration during the acute stages, progressing to squamous metaplasia during the reparative stages. Repair in these regions was essentially complete by 14 days PI. In the olfactory epithelium and the vomeronasal organ, there was interstitial edema with necrosis and exfoliation of epithelial cells and minimal to moderate inflammatory cell response during the acute stages. Residual reparative lesions were still evident in the olfactory epithelium, the columnar epithelium and neuroepithelium of the vomeronasal organ, and the nasolacrimal duct at 14 days PI. Viral antigen was demonstrated by immunohistochemistry in all regions during the acute stages of the disease, with the exception of the vomeronasal organ. In view of these findings, infections of the respiratory tract with viruses such as SDAV could have significant effects on functions such as olfaction and chemoreception for > or = 2 weeks postexposure in this species.


title: Treatment of pediatric diaphyseal femur fractures.

text: Methods of treating pediatric diaphyseal femur fractures are dictated by patient age, fracture characteristics, and family social situation. The recent trend has been away from nonsurgical treatment and toward surgical stabilization. The clinical practice guideline on pediatric diaphyseal femur fractures was undertaken to determine the best evidence regarding a number of different options for surgical stabilization. The recommendations address treatments that include Pavlik harness, spica casts, flexible intramedullary nailing, rigid trochanteric entry nailing, submuscular plating, and pain management. The guideline authors conclude that controversy and lack of conclusive evidence remain regarding the different treatment options for pediatric femur fractures and that the quality of scientific evidence could be improved for the revised guideline.


title: Australian Hajj pilgrims' knowledge, attitude and perception about Ebola, November 2014 to February 2015.

text: Upon return from Hajj 2014, 150 Australian pilgrims were interviewed about their understanding of the Ebola epidemic. Most (89%, 134/150) knew of the epidemic before travelling and 60% (80/134) of those knew Ebola transmits through body fluids. Pilgrims who received pre-travel health advice were more conscious of Ebola (69% vs 31%, p = 0.01) and adhered better to hand hygiene after touching an ill person (68% vs 31%, p < 0.01). Mass media was the main information source (78%).


title: The threat and prospects for control of an influenza pandemic.

text: Influenza constitutes the most widespread and significant respiratory infectious disease in the world, resulting in increased morbidity, mortality and economic loss each epidemic year. Pandemic influenza is a worldwide epidemic usually caused by a new virus variant to which the majority of the population has no immunity. As demonstrated in the devastating pandemic of 1918 to 1919, a pandemic virus may infect 30 to 50% of the worlds population and kill 1 to 2% of those infected. Pandemic control must be a concerted and co-ordinated world strategy and under the auspices of the World Health Organization, pandemic preparedness plans have been formulated, including: intensified surveillance for more rapid identification of new reassortant viruses with potential human virulence and infectivity, laboratory characterization of the new viruses so that vaccine may be prepared, development of techniques for more rapid vaccine production and the manufacture and stock piling of antiviral drugs. The H5N1 outbreak of virulent chicken influenza in 1997 in Hong Kong which resulted in the deaths of six of 18 infected persons serves as a wake-up call. Should such a virus attain high transmissibility in humans, a pandemic of tragic proportions might ensue. Even though the timing of onset of the next pandemic cannot be precisely predicted, world governments must understand the urgency of the problem and increase funding for influenza pandemic control.


title: [Monitoring of influenza and other respiratory diseases causative agents in children, hospitalized with community-acquired pneumonia in 2012-2013 epidemic season].

text: AIM Study the circulation of respiratory viruses in children with community-acquired pneumonia (CAP) during the period from October 2012 to May 2013. MATERIALS AND METHODS 136 children with CAP aged from 3 months to 16 years with ARI symptoms at the disease debut were studied. RNA/DNA of influenza A, B, parainfluenza (PI); adeno-, rhino-, RS-viruses, corona-, metapneumo- (MPV) and bocaviruses were detected in nasopharynx smears by PCR with hybridization-fluorescent detection in real time. Antibodies against influenza viruses A/H1N1/pdm09 California/07/09, epidemic reference strains of influenza viruses A/H1N1/Brisbane/59/07, A/ H3N2/Victoria/361/201 1, B/Wisconsin/1/10, against PI viruses type 1, 2, 3 were determined in paired sera by HAI. RESULTS In February-March 2013 the number of children protected by antibodies against influenza decreased, and circulation of influenza viruses A/H3N2 and A/H1N1/ pdm09 was detected. Rhinoviruses and PI viruses were determined throughout the epidemic season, bocavirus and adenoviruses--during the autumn-winter period, RS-virus and MPV--during winter-spring. Coronaviruses were not detected. The peak of virus detection was established in February when the threshold of influenza and ARI morbidity was exceeded. The main pathogens of children of the first 3 years of life are rhinoviruses, RS-virus, PI viruses and bocavirus. RS-virus infection at the debut of CAP in children younger than 3 years in 55.5% of cases is associated with the development of broncho-obstructive syndrome. Bocavirus infection in 50% of cases progresses with laryngo-tracheitis and bronchiolitis. CONCLUSION The fraction of viruses in etiologic structure ofARI in children varies depending on immune layer, season and age of children. Etiology of viral infection at the debut of CAP could only be proven using specialized laboratory studies.


title: Assessment of the inhibitory effect of ribavirin on the rainbow trout rhabdovirus VHSV by real-time reverse-transcription PCR.

text: Viral hemorrhagic septicemia virus (VHSV) is one of the most ubiquitous viruses in salmonid aquaculture in Europe. This infectious disease results in significant losses in the farming industry and therefore effective therapeutic agents are needed to control outbreaks caused by this pathogen. Thus, accurate methods to test new antiviral compounds need to be developed. Our goal was to establish a model system for testing novel antivirals with potential applications to aquaculture. In a previous study, a TaqMan real-time RT-PCR assay was designed to detect and quantitate VHSV in rainbow trout tissues [Chico, V., Gomez, N., Estepa, A., Perez, L., 2006. Rapid detection and quantitation of viral hemorrhagic septicemia virus in experimentally challenged rainbow trout by real-time RT-PCR. J. Virol. Methods 132, 154-159]. In this report, we applied the real-time RT-PCR assay to the evaluation of the inhibitory effect of ribavirin, a well-known broad spectrum antiviral drug, in a cell culture system. When added from the beginning of the infection, ribavirin caused a dose-dependent reduction of VHSV RNA accumulation. Real-time RT-PCR measurements showed 99.8% inhibition at 25 microg/ml ribavirin, with an IC50 of 0.43 microg/ml. Ribavirin maintained its inhibitory activity against VHSV when added at 6 h post-infection. Quantitation of N protein messenger RNA and plus-stranded RNA showed a substantial decrease of viral transcription in ribavirin-treated cells. Partial reversion of the effect of ribavirin by addition of GTP was observed, confirming that ribavirin targets the synthesis of guanidine nucleotides in the cells. This is the first report of a real-time PCR-based assay for addressing the efficacy and mechanism of action of an antiviral agent for rainbow trout.


title: [Evaluation of noninvasive positive pressure ventilation in treatment for patients with severe acute respiratory syndrome].

text: OBJECTIVE To analyze the effects of noninvasive positive pressure ventilation (NIPPV) on oxygenation of severe acute respiratory syndrome (SARS) patients, and to discuss the timing point for mechanical ventilation. METHODS Twenty-five SARS patients with respiratory dysfunction treated with NIPPV were studied retrospectively in order to evaluate the influences within 24 hours after initiation of ventilatory support on their physiological indices and oxygenation. Patients with SARS were divided into two groups: survivor group (n=13) and non-survivor group (n=12). We compared the acute physiology and chronic health evaluation (APACHEII) score, respiratory rate (RR),saturation of oxygen (SpO(2)) and modificative respiratory index (MRI) for the survivors and non-survivors before NIPPV and after NIPPV for twenty-four hours, respectively. RESULTS Although NIPPV administered via full-face masks might be an effective treatment for rapidly improving vital signs and gas exchange and sense of dyspnea in both groups during the initial 24 hours of ventilatory support, the patients in non-survivor group had higher APACHEII score, respiratory rates and lower SpO(2), MRI than the patients in survivor group (P<0.05) at the same intervals after initiation of support. CONCLUSION Noninvasive ventilation should be used as a substitutive tool for endotracheal intubation an alternative treatment for acute respiratory failure related to SARS. Therefore, we should make efforts to avoid missing the time point for NIPPV or intubation, and we should not be restricted to the available indications for NIPPV or IPPV.


title: Protection from infection with severe acute respiratory syndrome coronavirus in a Chinese hamster model by equine neutralizing F(ab')2.

text: To warrant potential clinical testing, the equine anti-severe acute respiratory syndrome coronavirus (SARS-CoV) F(ab')(2) requires evaluation in as many animal models as possible. In this study, we established a new animal model, the Chinese hamster, susceptible to SARS-CoV infection. SARS-CoV could propagate effectively and sustain high levels for 1 wk in animal lungs. All animals were protected from SARS-CoV infection in preventive settings. Further, when used therapeutically this antibody led to an approximately 4-log(10) decrease in viral burden in infected animal lungs. The pathological changes in lungs correlated closely with the dose of antibody administered. The excellent preventive and therapeutic roles of equine anti-SARS-CoV F(ab')(2) in several animal models, including the novel Chinese hamster model described in this study, have provided exciting data concerning its potential clinical study.


title: A novel glycoprotein of feline infectious peritonitis coronavirus contains a KDEL-like endoplasmic reticulum retention signal.

text: A new protein of feline infectious peritonitis coronavirus (FIPV) was discovered in lysates of [35S]cysteine-labeled infected cells. Expression of open reading frame (ORF) 6b of FIPV in recombinant vaccinia virus-infected cells was used to identify it as the 6b protein. Further characterization revealed that it is a novel type of viral glycoprotein whose function is not clear. It is a soluble protein contained in microsomes; its slow export from the cell is caused by the presence of an endoplasmic reticulum (ER) retention signal at the C terminus. This amino acid sequence, KTEL, closely resembles the consensus KDEL signal of soluble resident ER proteins. A mutant 6b protein with the C-terminal sequence KTEV became resistant to digestion by endo-beta-N-acetylglucosaminidase H with a half-time that was reduced threefold. In contrast, a mutant with the sequence KDEL was completely retained in the ER. The FIPV 6b protein is the first example of a viral protein with a functional KDEL-like ER retention signal.


title: Interim results: state-specific seasonal influenza vaccination coverage - United States, August 2009-January 2010.

text: The advent of the 2009 influenza A (H1N1) pandemic in April 2009 made the 2009-10 influenza season highly unusual. Public awareness of the potential seriousness of influenza was heightened by media coverage of pandemic-associated hospitalizations and deaths, especially among younger persons. In the fall, the distribution of two separate influenza vaccines began, with distinct, although overlapping, recommendations from the Advisory Committee on Immunization Practices (ACIP). In addition, 2009-10 was the first full season in which ACIP's recommendation to vaccinate all children aged 5--18 years was implemented. To provide preliminary state-specific estimates of seasonal influenza vaccination coverage, CDC analyzed Behavioral Risk Factor Surveillance System (BRFSS) and National 2009 H1N1 Flu Survey (NHFS) data collected during October 2009-February 2010. By January 31, estimated state seasonal influenza vaccination coverage among persons aged >or=6 months ranged from 30.3% to 54.5% (median: 40.6%). Median coverage was 41.2% for children aged 6 months-17 years, 38.3% for adults aged 18--49 years with high-risk conditions, 28.8% for adults aged 18-49 years without high-risk conditions, 45.5% for adults aged 50-64 years, and 69.3% for adults aged >or=65 years. These results, compared with the previous season, suggest large increases in coverage for children and a moderate increase for adults aged 18-49 years without high-risk conditions. Health departments should identify best practices that lead to higher vaccination coverage and should support effective vaccination services (e.g., school-located vaccination programs and office-based protocols, such as reminder/recall and standing orders).


title: Serological studies in Austrian fattening pigs with respiratory disorders.

text: Serum samples of 253 fattening pigs out of 44 different herds with respiratory signs were examined for antibodies to Aujeszky's disease virus by ELISA, swine influenza virus (HI test), porcine respiratory coronavirus (ELISA) and porcine reproductive and respiratory syndrome virus (IPMA). One single case of Aujeszky's disease was detected at slaughter. On the other hand 24.5% of the animals proved to be positive for swine influenza, although no vaccine is licensed in Austria, and 63.6% reacted positively to porcine respiratory coronavirus. None of the sera showed antibodies to porcine reproductive and respiratory syndrome virus, which has not as yet been identified in the Austrian swine population.


Serologic survey of canine coronavirus in wild coyotes in the western United States, 1972-1982.


title: [Clinical application of radionuclide bone scintigraphy in diagnosis of avascular osteonecrosis in patients with severe acute respiratory syndrome in convalescence].

text: OBJECTIVE To evaluate the clinical value of radionuclide bone scintigraphy in diagnosis of avascular osteonecrosis in patients with severe acute respiratory syndrome (SARS) in convalescence. METHODS We performed three-phase bone scintigraphy of femoral head regions and whole-body bone scan in SARS patients 4-6 months after they recovered from the syndrome, and then compared the results with simultaneous MRI. RESULTS Typical avascular necrosis at different stages and severities was found on bone scintigraphy at 31 femoral heads of 16 SARS patients, 97% of which were MRI positive. Suspicious necrosis was found at 42 femoral heads of 23 patients, 67% of which were MRI negative. Among 30 patients with normal three-phase scintigraphic results, 10% of whom were suspicious on MRI. In addition, abnormal distributions of radioactivity were observed in other bones on the whole-body bone scans of 29 patients, including osteonecrosis of knees in 15 patients. CONCLUSIONS Radionuclide bone scintigraphy is valuable in early diagnosis of osteonecrosis in SARS patients in convalescence. It provides a mutually supplementary tool for MRI.


title: Single incision laparoscopic cholecystectomy--a new advantage of gallbladder surgery.

text: In this study is demonstrated our experience in single incision laparoscopic cholecystectomy (SILS), compared to standard laparoscopic cholecystectomy. There were 48 single incision laparoscopic cholecystectomies (SILS) performed during one-year period (A group) and results have been compared with a group of 50 patients who underwent standard laparoscopic cholecystectomy (B group). Outcome measures included operative time, need for conversion, complications, additional analgesia for pain control after procedure, hospital stay and cosmetic outcome. The mean operative time was 46 +/- 3.5 min in A group, and 43 +/- 4 min in B patients group. Early postoperative complications were not detected. The mean hospitalization period was 2 days in both groups. Our experience suggests that SILS cholecystectomy can be performed with outcome similar to standard laparoscopic surgery while affording better cosmesis.


SARS could still affect the United Kingdom, health secretary warns.


title: Identification and characterization of fully human anti-CD22 monoclonal antibodies.

text: CD22 is a member of the B cell receptor family and is implicated in B cell function and development. It is expressed on multiple forms of B cell lymphoma and is an attractive cancer therapeutic target. We report here the identification of two fully human anti-CD22 antibodies using phage display methodology. Both antibodies exhibit specific binding to cell surface-associated CD22 in multiple B cell lines. Through ELISA using mammalian cell-expressed sub-domains of CD22 as binding antigen, we mapped the binding epitopes of the newly identified CD22 antibodies to be within the Ig-like domains 5 to 7 of CD22. Their epitopes do not overlap with those of several therapeutic antibodies currently in preclinical or clinical development. These antibodies have potential as cancer therapeutic candidates and research reagents.


title: Human infection by avian influenza A H5N1.

text: The Southeast Asian outbreak of the highly lethal avian influenza A H5N1 infection in humans is unlikely to abate because of the enormous number of backyard farms providing poultry as the main source of food protein in developing countries. This increases the risk of the emergence of a reassortant pandemic influenza virus with improved human-to-human transmissibility. Currently triage of suspected cases by epidemiological risk factors remains the only practical way of case identification for laboratory investigation and infection control. The clinical usefulness of rapid diagnostic laboratory tests requires more vigorous evaluation. The lethality of this disease may reflect systemic viral dissemination, cytokine storm, or alveolar flooding due to inhibition of cellular sodium channels. The present circulating genotype Z is intrinsically resistant to amantadine and rimantadine. Prognosis may be improved by early treatment with a neuraminidase inhibitor with good systemic drug levels, and post-exposure prophylaxis for health care workers is recommended. The role of immunomodulators and other modalities of therapy requires evaluation in randomised controlled trials, with prospective monitoring of the viral load and cytokine profiles in various clinical specimens. In view of the high fatality of the disease, a combination of contact, droplet, and airborne precautions are recommended as long as resources allow despite the fact that the relative importance of these three modes in nosocomial transmission of avian influenza is still unknown.


title: Mutagenic analysis of the coronavirus intergenic consensus sequence.

text: Previously, a system in which an intergenic region from mouse hepatitis virus (MHV) inserted into an MHV defective interfering (DI) RNA led to transcription of a subgenomic DI RNA in helper virus-infected cells was established. In the present study, a DI cDNA containing one UCUAAAC consensus sequence in the middle of the 0.3-kb-long intergenic region located between genes 6 and 7 was constructed. From this DI cDNA clone, 21 mutant DI RNAs were constructed so that each of the seven consensus sequence nucleotides was changed individually to the three alternative bases. These mutants were used to define how changes in the integrity of MHV transcription consensus sequence UCUAAAC affected mRNA transcription. Except for two mutants with the sequences UGUAAAC and UCGAAAC, all of the mutants supported efficient subgenomic DI RNA transcription. This indicated that MHV transcription regulation was sufficiently flexible to recognize altered consensus sequences. Next, these and other mutants were used to examine the leader-body fusion site on the subgenomic DI RNAs. Sequence analysis demonstrated that all subgenomic DI RNAs analyzed contained two pentanucleotide sequences; the first sequence seemed to be contributed by the leader, and the leader-body fusion most likely took place at either the first or the second nucleotide of the second sequence. This observation was not consistent with the proposed coronavirus transcription model (S. C. Baker and M. M. C. Lai, EMBO J. 9:4173-4179, 1990) which states that nucleotide mismatch can be corrected by RNA polymerase proofreading activity.


Continuous transcutaneous monitoring.


title: Drug targets for rational design against emerging coronaviruses.

text: The recent, fatal outbreak of the novel coronavirus strain in the Middle East highlights the real threat posed by this unique virus family. Neither pharmaceutical cures nor preventive vaccines are clinically available to fight against coronavirus associated syndromes, not to mention a lack of symptom soothing drugs. Development of treatment options is complicated by the unpredictable, recurring instances of cross-species viral transmission. The vastly distributing virus reservoir and the rapid rate of host-species exchange of coronavirus demands wide spectrum potency in an ideal therapeutic. Through summarizing the available information and progress in coronavirus research, this review provides a systematic assessment of the potential wide-spectrum features on the most popular drug targets including viral proteases, spike protein, RNA polymerases and editing enzymes as well as host-virus interaction pathways associated with coronaviruses.


title: [Detection and sequence analysis of an imperfect stem-loop structure in cis activating gene element from SV40PolyA].

text: Our previous studies showed that tandem Alu repeats inhibited GFP gene expression when they were inserted into the downstream of GFP gene in pEGFP-C1 vector and HeLa cells were then transfected transiently. The sequence named 2F2R (second 60 bp from the 5' end of SV40PolyA antisense strand) eliminated the repression of GFP gene expression induced by Alu repeats when 2F2R was inserted between GFP and Alu repeats. In this study the deletion of 2F2R DNA showed that 45R (45 bp in 2F2R 5'end), 30R (30 bp in 2F2R 5' end) and 22R (22 bp in 2F2R 5' end) activated GFP gene expression, and the activating actions of the double tandem sequences were stronger than those of their corresponding single sequences. Secloop (22 bp near the center in 2F2R) and Poly4 (30 bp in 2F2R 3' end) sequences did not activate GFP gene expression. The activating action of 30R-Poly4 sequence formed by ligating 30R with Poly4 by 9 bp was lower than that of 2F2R. The linking base number between two 22R sequences did not influence the GFP gene expression obviously. Sequence 22R (5'-GTGAAAAAAATGCTTTATTTGT-3') contains an imperfect palindrome sequence and may form an imperfect stem-loop structure including a 3nt loop, 3 bp first stem, 2nt bulge, and 3bp second stem. The mutations changing stem-loop structure of 22R influenced the GFP gene activation significantly and neither the excessively stable nor excessively unstable stem-loop structures were in favour of GFP gene activation, which suggested that the suitably imperfect stem-loop structures had something with gene activation.


An iatrogenic pandemic of panic.


title: [COVID-19: patient zero in the Netherlands].

text: BACKGROUND Since December 2019, the world is captivated by SARS-CoV-2, a new coronavirus that shows a lot of similaritieswith previous coronaviruses such as SARS and MERS. Although it was initially seen mainly in China and the surrounding countries, now it also reached Europe, where a large region in northern Italy, in particular, encountered many infections. CASE DESCRIPTION Here we describe the first Dutch patient with COVID-19, a 56-year-old man whose infection appeared to be related to a trip to Northern Italy one week before presentation. In the days that followed, the brother of the patient with whom he had traveled, his wife and daughter also tested positive. CONCLUSION At the moment much is still unclear and it is particularly important to quickly identify patients with an increased risk of complications and to prevent unrestrained spread in the Netherlands.


Heroes and heroines of the war on SARS.


title: [Syndromic surveillance: a finger on the pulse of public health].

text: Syndromic surveillance is a monitoring methodology in which data from disease indicators are collected and interpreted as quickly as possible (in real time) to get an impression of the current morbidity in the population. In this case 'syndrome' means: behavior, signs and symptoms that may be indicative of an infection, without a proven pathogen, or incidents such as an environmental disaster. Syndromic surveillance has gained momentum in response to various international infectious disease threats, including the outbreak of severe acute respiratory syndrome (SARS) in 2003. In several European countries, syndromic surveillance has been used to monitor the impact of outbreaks or natural disasters and the health of visitors of mass events. Following the outbreak of an infectious disease, syndromic surveillance can be reassuring in the absence of an epidemic. Insight into burden of disease can provide guidance in adjusting care capacity to need. Several syndromic surveillance systems and pilot studies have been set up recently in the Netherlands. The costs, benefits and their best application should be evaluated in the near future.


title: Proportion of workers who were work-injured and payment by workers' compensation systems - 10 states, 2007.

text: Work-related injuries are a major cause of morbidity in the United States, with approximately 4 million employer-reported nonfatal injuries and illnesses in 2007. Currently, few population-based state-level estimates of nonfatal occupational injury rates exist. In the few extant studies, self-reported, nonfatal occupational injury rates exceed estimates based on employer reports or state workers' compensation systems. To estimate the proportion of workers who were work-injured during the preceding 12 months and the proportion of those injured for whom workers' compensation insurance programs paid for medical care, 10 states added a module to their 2007 Behavioral Risk Factor Surveillance System (BRFSS) survey. This report summarizes the results of that survey, which found that the proportion of workers who were work-injured during the preceding 12 months ranged from 4.0 to 6.9 work-injured persons per 100 employed persons (Kentucky and New York, respectively). The proportion of self-reported work-injured persons for whom medical treatment was paid by workers' compensation insurance ranged from 47% in Texas to 77% in Kentucky. This study shows the feasibility of complementing existing occupational injury surveillance through the use of population-based surveys. States that wish to enhance existing occupational injury surveillance should consider similar studies. Additional research is needed to understand the reasons for nonpayment of worker-reported occupational injuries by workers' compensation insurance programs.


title: Vaccinia virus leads to ATG12–ATG3 conjugation and deficiency in autophagosome formation.

text: The interactions between viruses and cellular autophagy have been widely reported. On the one hand, autophagy is an important innate immune response against viral infection. On the other hand, some viruses exploit the autophagy pathway for their survival and proliferation in host cells. Vaccinia virus is a member of the family of Poxviridae which includes the smallpox virus. The biogenesis of vaccinia envelopes, including the core envelope of the immature virus (IV), is not fully understood. In this study we investigated the possible interaction between vaccinia virus and the autophagy membrane biogenesis machinery. Massive LC3 lipidation was observed in mouse fibroblast cells upon vaccinia virus infection. Surprisingly, the vaccinia virus induced LC3 lipidation was shown to be independent of ATG5 and ATG7, as the atg5 and atg7 null mouse embryonic fibroblasts (MEFs) exhibited the same high levels of LC3 lipidation as compared with the wild-type MEFs. Mass spectrometry and immunoblotting analyses revealed that the viral infection led to the direct conjugation of ATG3, which is the E2-like enzyme required for LC3-phosphoethanonamine conjugation, to ATG12, which is a component of the E3-like ATG12–ATG5-ATG16 complex for LC3 lipidation. Consistently, ATG3 was shown to be required for the vaccinia virus induced LC3 lipidation. Strikingly, despite the high levels of LC3 lipidation, subsequent electron microscopy showed that vaccinia virus-infected cells were devoid of autophagosomes, either in normal growth medium or upon serum and amino acid deprivation. In addition, no autophagy flux was observed in virus-infected cells. We further demonstrated that neither ATG3 nor LC3 lipidation is crucial for viral membrane biogenesis or viral proliferation and infection. Together, these results indicated that vaccinia virus does not exploit the cellular autophagic membrane biogenesis machinery for their viral membrane production. Moreover, this study demonstrated that vaccinia virus instead actively disrupts the cellular autophagy through a novel molecular mechanism that is associated with aberrant LC3 lipidation and a direct conjugation between ATG12 and ATG3.


title: Confronting an influenza pandemic: ethical and scientific issues.

text: The prolonged concern over the potential for a global influenza pandemic to cause perhaps many millions of fatalities is a chilling one. After the SARS (severe acute respiratory syndrome) scares [1], attention has turned towards the possibility of an avian influenza virus hybridizing with a human influenza virus to create a highly virulent, as yet unknown, killer, on a scale unseen since the Spanish flu outbreak of 1918, which produced more fatalities than the Great War. In deciding how countries should react to this potential pandemic, individually and collectively, a reasonable and practical balance must be struck between the rights and obligations of individual citizens and protection of the wider community and, indeed, society as a whole. In this communication, ethical issues are discussed in the context of some of the scientific questions relating to a potential influenza pandemic. Among these issues are the rights and obligations of healthcare professionals, difficulties surrounding resource allocation, policies that have an impact on liberty and trade, when and how to introduce any vaccine or other form of mass treatment, global governance questions and the role of health policies in contemporary society. By considering these issues and questions in advance of an influenza, or indeed any other, pandemic commencing, countries can be better prepared to deal with the inevitably difficult decisions required during such events, rather than dusting down outdated previous plans, or making and implementing policy in an ad hoc manner with a resultant higher risk of adverse consequences.


title: Comparative evaluation of two severe acute respiratory syndrome (SARS) vaccine candidates in mice challenged with SARS coronavirus.

text: Two different severe acute respiratory syndrome (SARS) vaccine strategies were evaluated for their ability to protect against live SARS coronavirus (CoV) challenge in a murine model of infection. A whole killed (inactivated by beta-propiolactone) SARS-CoV vaccine and a combination of two adenovirus-based vectors, one expressing the nucleocapsid (N) and the other expressing the spike (S) protein (collectively designated Ad S/N), were evaluated for the induction of serum neutralizing antibodies and cellular immune responses and their ability to protect against pulmonary SARS-CoV replication. The whole killed virus (WKV) vaccine given subcutaneously to 129S6/SvEv mice was more effective than the Ad S/N vaccine administered either intranasally or intramuscularly in inhibiting SARS-CoV replication in the murine respiratory tract. This protective ability of the WKV vaccine correlated with the induction of high serum neutralizing-antibody titres, but not with cellular immune responses as measured by gamma interferon secretion by mouse splenocytes. Titres of serum neutralizing antibodies induced by the Ad S/N vaccine administered intranasally or intramuscularly were significantly lower than those induced by the WKV vaccine. However, Ad S/N administered intranasally, but not intramuscularly, significantly limited SARS-CoV replication in the lungs. Among the vaccine groups, SARS-CoV-specific IgA was found only in the sera of mice immunized intranasally with Ad S/N, suggesting that mucosal immunity may play a role in protection for the intranasal Ad S/N delivery system. Finally, the sera of vaccinated mice contained antibodies to S, further suggesting a role for this protein in conferring protective immunity against SARS-CoV infection.


title: Cell surface phenotype and human T lymphotropic virus type 1 antigen expression in 12 T cell lines derived from peripheral blood and cerebrospinal fluid of West Indian, Guyanese and African patients with tropical spastic paraparesis.

text: Twelve long-term cell lines were established from peripheral blood mononuclear cells (PBMC) or cerebrospinal fluid cells of patients with human T lymphotropic virus type I (HTLV-1) seropositive tropical spastic paraparesis (TSP) originating from the French West Indies, French Guyana or the Central African Republic. Most of these long-term interleukin-2-dependent cell lines exhibited a pattern characteristic of CD4(+)-activated T cells with high expression of CD2, CD3 and CD4 antigens, associated with a strong density of TAC and DR molecules. Nevertheless, in five cases CD8 expression was present at a significant level. HTLV-I antigens were never detected in uncultured PBMC, but they were expressed in a few cells after short-term culture and after 4 months the majority of the cells were HTLV-I positive, as demonstrated by indirect immunofluorescence (IF) using polyclonal or monoclonal anti-p19 and anti-p24 antibodies. Low and variable levels of reverse transcriptase activity were detected in supernatant fluids of these cell lines only after 4 months of culture, when at least 50% of the cells exhibited HTLV-I antigens by IF. However, numerous type C HTLV-I-like viral particles were detected, mostly in the extracellular spaces, with rare budding particles. Similar findings were found in three T cell lines derived from West Indian and African patients with adult T-cell leukaemia/lymphoma (ATLL). Differences in high Mr polypeptides were detected by Western blot in cell lysates when comparing TSP- or ATLL-derived T cell lines. Thus a signal of 62K was easily detectable in all the TSP lines, but not in the ATLL lines. In all cell lines bands corresponding to p53, p24 and p19 viral core polypeptides were present, as was the env gene-coded protein p46.


title: Gene profiles during root canal treatment in experimental rat periapical lesions.

text: The purpose of this study was to profile gene expression in periapical lesions during root canal treatment (RCT). Periapical lesions were induced experimentally by exposing the pulp in Sprague-Dawley rats. After 3 wk, the animals received root canal filling (RCF) and were sacrificed 1 or 4 wk later. From the periapical tissues, total RNA was extracted and processed for cDNA-microarray analysis. The lesions were histologically and radiographically confirmed to expand 4 wk after pulp exposure (inflammation phase) and to stabilize 4 wk after RCF (healing phase). In approximately 30,000 genes on the microarray, 203 genes were up-regulated to more than 5-fold (e.g., IL-1beta), and 864 genes were down-regulated to less than 20% of baseline level (e.g., caspase 8) in inflammation phase. Compared with inflammation phase, we found that 133 genes were up-regulated (e.g., IL-1alpha) and 50 genes were down-regulated (e.g., defensin alpha5) in healing phase. Corresponding to the gene expression profiles, accumulation of IL-1alpha and IL-1beta was observed in the periapical lesions by immunohistochemistry. These gene profiles might be useful in diagnosing the healing process of periapical lesions.


title: An outbreak of meningococcal disease in a secondary school--implications for public health practice.

text: This report describes briefly the management of three cases of meningococcal disease which all occurred within one week at a secondary school on the Central Coast of New South Wales in late winter 2003. The Central Coast health area has a population of approximately 300,000. Between 10 and 15 cases of meningococcal disease are notified to the Central Coast Public Health Unit each year. The three cases all presented to Gosford Hospital, Cases 1 and 2, both in Year 9, on Thursday 14 August 2003 and Case 3 in Year 8 on Friday 15 August 2003.


title: Dissection of Extracranial Internal Carotid Artery Due to Balloon Guiding Catheter Resulting in Asymptomatic Internal Carotid Artery Occlusion.

text: Dissection of the internal carotid artery (ICA) is a rare condition that accounts for a significant proportion of ischemic strokes in young adults. Iatrogenic dissection as a complication of neurointerventional procedures is a traumatic dissection which has been reported relatively rare in the literature. In this report, a case of dissection of the ICA is reported that was caused by repetitive movement of the balloon guiding catheter during stent-assisted thrombectomy (SAT), resulting in occlusion of the ICA.


title: Current developments in hernia repair; meshes, adhesives, and tacking.

text: Open and laparoscopic hernia surgery continues to evolve with new products allowing surgeons multiple choices in treating their patients. The evolution towards tension-free techniques in dealing with hernias requires that today's surgeons know the options available in meshes as well as fixation methods in order to have the best outcomes. In recent years, there has been a rapid expansion in the number of meshes available. Currently, there are numerous uncoated, coated, and biologic meshes in production that can be used in hernia repair. This paper will focus on the latest developments in coated meshes that allow for intra-abdominal placement as well as the different types of biologic meshes and their typical uses. Tacking devices for laparoscopic hernia repair now come in titanium as well as absorbable devices. AbsorbaTack™ (Covidien, Norwalk, CT) and Sorbafix™ (Davol, Warwick, RI) are two of the newest absorbable tacking devices thought to possibly benefit patients with decreased pain and long-term complications as compared with their titanium counterparts. Adhesives continue to be used more and more for hernia repair, especially in inguinal and paraesophageal hernia repairs. Tissucol™/Tisseel™ (Baxter, Deerfield, IL) and Evicel™ (Ethicon, Somerville, NJ) are two types of fibrin glues that are available for use in hernia repair. Practitioners using these biologic adhesives think there is less pain compared with tacking.


title: High-sensitivity detection methods for low-abundance RNA species: applications for functional genomics research.

text: Gene expression analysis has facilitated a more complete understanding of the molecular biology of cellular processes and how variations of RNA expression are useful for the classification of various diseases. Furthermore, recent analysis of a variety of noncoding RNAs, such as microRNAs, has demonstrated that these RNAs play an important role in many cellular events, including cell differentiation and death, and may also serve as biological markers for disease. Besides helping in the understanding of diseases, RNA analysis is used in drug discovery, patient prognosis and treatment evaluation. One obstacle left to overcome is the amount of material required for the analysis, particularly when trying to extract information from precious, limited, clinical samples. Here we review the many approaches scientists take to either amplify the amount of RNA or amplify the signal generated from small amounts of RNA.


title: [SARS (Severe Acute Respiratory Syndrome). Emergent transmissible disease].

text: Of the reemergent transmissible diseases of the past decades, SARS is probably not the last to express the alterations occurring in the relationships of the human being with its global ecosystem. The life of contemporary man is characterized, among others, by a huge thirst for traveling, for varied reasons, consequence of the globalization process. SARS virus, mutant belonging to Coronaviridae, occurred in one of the most densely populated areas of the world. There are two main moments marking the reemergence and evolution of SARS: firstly, the onset of the epidemic in China in November 2002 followed by the worldwide spread of the epidemiological process, and secondly the discovery of SARS virus as a mutant of coronaviruses in March-April 2003 in USA, Canada, and Hong Kong. The possibilities of general and special prevention, and particularly vaccine prevention are likely to bring this disease under control.


title: Severe acute respiratory syndrome: clinical and laboratory manifestations.

text: Severe acute respiratory syndrome (SARS) is a recently emerged infectious disease with significant morbidity and mortality. An epidemic in 2003 affected 8,098 patients in 29 countries with 774 deaths. The aetiological agent is a new coronavirus spread by droplet transmission. Clinical and general laboratory manifestations included fever, chills, rigor, myalgia, malaise, diarrhoea, cough, dyspnoea, pneumonia, lymphopenia, neutrophilia, thrombocytopenia, and elevated serum lactate dehydrogenase (LD), alanine aminotransferase (ALT) and creatine kinase (CK) activities. Treatment has been empirical; initial potent antibiotic cover, followed by simultaneous ribavirin and corticosteroids, with or without pulse high-dose methylprednisolone, have been used. The postulated disease progression comprises (1) active viral infection, (2) hyperactive immune response, and (3) recovery or pulmonary destruction and death. We investigated serum LD isoenzymes and blood lymphocyte subsets of SARS patients, and found LD1 activity as the best biochemical prognostic indicator for death, while CD3+, CD4+, CD8+ and natural killer cell counts were promising predictors for intensive care unit (ICU) admission. Plasma cytokine and chemokine profiles showed markedly elevated Th1 cytokine interferon (IFN)-gamma, inflammatory cytokines interleukin (IL)-1beta, IL-6 and IL-12, neutrophil chemokine IL-8, monocyte chemoattractant protein-1 (MCP-1), and Th1 chemokine IFN-gamma-inducible protein-10 (IP-10) for at least two weeks after disease onset, but there was no significant elevation of inflammatory cytokine tumor necrosis factor (TNF)-alpha and anti-inflammatory cytokine IL-10. Corticosteroid reduced IL-8, MCP-1 and IP-10 concentrations from 5-8 days after treatment. Measurement of biochemical markers of bone metabolism demonstrated significant but transient increase in bone resorption from Day 28-44 after onset of fever, when pulse steroid was most frequently given. With tapering down of steroid therapy, there was a decrease in bone resorption marker together with an increase in bone formation markers round Day 50, suggesting that some of the bone loss might be reversed. Our research studies on the chemical pathology and clinical immunology of SARS should have implications for the pathophysiology and therapy of this potentially lethal infection.


title: Recommendations for a standardised educational program in robot assisted gynaecological surgery: Consensus from the Society of European Robotic Gynaecological Surgery (SERGS).

text: Background The Society of European Robotic Gynaecological Surgery (SERGS) aims at developing a European consensus on core components of a curriculum for training and assessment in robot assisted gynaecological surgery. Methods A Delphi process was initiated among a panel of 12 experts in robot assisted surgery invited through the SERGS. An online questionnaire survey was based on a literature search for standards in education in gynaecological robot assisted surgery. The survey was performed in three consecutive rounds to reach optimal consensus. The results of this survey were discussed by the panel and led to consensus recommendations on 39 issues, adhering to general principles of medical education. Results On review there appeared to be no accredited training programs in Europe, and few in the USA. Recommendations for requirements of training centres, educational tools and assessment of proficiency varied widely. Stepwise and structured training together with validated assessment based on competencies rather than on volume emerged as prerequisites for adequate and safe learning. An appropriate educational environment and tools for training were defined. Although certification should be competence based, the panel recommended additional volume based criteria for both accreditation of training centres and certification of individual surgeons. Conclusions Consensus was reached on minimum criteria for training in robot assisted gynaecological surgery. To transfer results into clinical practice, experts recommended a curriculum and guidelines that have now been endorsed by SERGS to be used to establish training programmes for robot assisted surgery.


title: What role for human rhinoviruses in the lower respiratory tract?

text: Human rhinoviruses (HRV) usually cause upper airway infections. However, viral replication in the tracheobronchial tree has been disclosed, although its clinical role is poorly known. We evaluated the prevalence of HRV in 159 bronchoalveolar lavages from 88 patients and describe a lung transplant recipient with a high HRV load in association with acute rejection. HRV was detected in 22/88 patients (25.0%): 7/18 lung transplant recipients, 11/41 immunocompetent, and 4/29 immunocompromised (p = n.s.). No lung disease was significantly associated with HRV positivity. It should be recommended to include HRV in the virological diagnostic work-up of lower respiratory specimens to elucidate their role.


title: Elicitation of immunity in mice after immunization with the S2 subunit of the severe acute respiratory syndrome coronavirus.

text: The S2 domain of the severe acute respiratory syndrome coronavirus (SARS-CoV) spike (S) protein is responsible for fusion between virus and target cell membranes, and is expected to be immungenic. In this study, we investigated the immune responses against the S2 subunit in BALB/c mice, which were vaccinated either with plasmid DNA encoding the S2 domain (residues 681-1120), the recombinant S2 fragment (residues 681-980) in incomplete Freund's adjuvant, or with inactivated SARS-CoV. The increased number of specific cytotoxic cells (CTLs) and the high titer of specific antibody showed stimulation of both arms of the immune system in these groups. The shift in cytokines suggested that Th1-polarized immune response was induced by plasmid pCoVS2, meanwhile the Th2-dominant response was induced by recombinant S2 fragment and inactivated vaccine. However, the titer of neutralizing antibodies was only detectable in mice immunized with inactivated virus, but not with pCoVS2 plasmid. Taken together, the S2 domain could induce specific cellular immune response and a high level of total IgG but little neutralizing antibodies against infection by SARSCoV.


title: Laparoscopic sleeve gastrectomy for morbid obesity with natural orifice specimen extraction (NOSE).

text: OBJECTIVE An experience with laparoscopic sleeve gastrectomy using the natural orifice specimen extraction (NOSE) technique. BACKGROUND Bariatric surgery is nowadays the only long term effective obesity treatment method. METHODS Twenty one consecutive patients underwent laparoscopic sleeve gastrectomy with the use of natural orifice specimen extraction (NOSE) in the Surgical Clinic of Faculty Hospital Ostrava between May 2012 and August 2012. Inclusion criteria were the body mass index (BMI) higher than 35 kg/m2 or higher than 32 kg/m2 accompanied with relevant comorbidities. RESULTS Among 21 patients in this series, there were three men (14.3%) and 18 women (85.7%). Their mean age was 40.9±10.2 years. Their mean preoperative BMI was 40.4±4.6 kg/m2. No patient had previous bariatric surgery, one patient had laparoscopic fundoplication. All operations were completed laparoscopically with no conversions to an open procedure. In two cases, laparoscopic cholecystectomy was performed and the gallbladder was extracted along with the gastric specimen by transgastric approach. CONCLUSION Laparoscopic sleeve gastrectomy is a safe and effective bariatric procedure with low morbidity and mortality. Based on our initial experiences it could be an indication for NOSE with transgastric approach. Obese patients would benefit from this approach due to the elimination of wound complications (Tab. 2, Fig. 3, Ref. 22).


title: The 2016 Infusion Therapy Standards of Practice.

text: Approximately every 5 years, the Infusion Nurses Society publishes evidence-based practice standards. This article provides an overview of the process used in standards development, describes the format of the standards, and provides a short summary of selected standards as applied to home care. The Standards are an important document that should be available to every home care organization that provides home infusion therapy.


title: Molecular evolution analysis and geographic investigation of severe acute respiratory syndrome coronavirus-like virus in palm civets at an animal market and on farms.

text: Massive numbers of palm civets were culled to remove sources for the reemergence of severe acute respiratory syndrome (SARS) in Guangdong Province, China, in January 2004, following SARS coronavirus detection in market animals. The virus was identified in all 91 palm civets and 15 raccoon dogs of animal market origin sampled prior to culling, but not in 1,107 palm civets later sampled at 25 farms, spread over 12 provinces, which were claimed to be the source of traded animals. Twenty-seven novel signature variation residues (SNVs) were identified on the spike gene and were analyzed for their phylogenetic relationships, based on 17 sequences obtained from animals in our study and from other published studies. Analysis indicated that the virus in palm civets at the live-animal market had evolved to infect humans. The evolutionary starting point was a prototype group consisting of three viral sequences of animal origin. Initially, seven SNV sites caused six amino acid changes, at positions 147, 228, 240, 479, 821, and 1080 of the spike protein, to generate low-pathogenicity viruses. One of these was linked to the first SARS patient in the 2003-2004 period. A further 14 SNVs caused 11 amino acid residue changes, at positions 360, 462, 472, 480, 487, 609, 613, 665, 743, 765, and 1163. The resulting high-pathogenicity groups were responsible for infections during the so-called early-phase epidemic of 2003. Finally, the remaining six SNVs caused four amino acid changes, at positions 227, 244, 344, and 778, which resulted in the group of viruses responsible for the global epidemic.


title: Expression of the recombinant anchorless N-terminal domain of mouse hepatitis virus (MHV) receptor makes hamster of human cells susceptible to MHV infection.

text: Mouse hepatitis virus (MHV) receptor, the receptor for the murine coronavirus MHV, was expressed in MHV-resistant hamster and human cells as a series of mutant, recombinant glycoproteins with carboxy-terminal deletions lacking the cytoplasmic tail, transmembrane domain, and various amounts of the immunoglobulin constant-region-like domains. The soluble receptor glycoproteins containing the N-terminal virus-binding domain were released into the supernatant medium and inactivated the infectivity of MHV-A59 virions in a concentration-dependent manner. Surprisingly, some of the anchorless glycoproteins were found on the plasma membranes of transfected cells by flow cytometry, and these cells were rendered susceptible to infection with three strains of MHV. Thus, in the cells in which the anchorless, recombinant receptor glycoprotein is synthesized, some of the protein is bound to an unidentified moiety on the plasma membrane, which allows it to serve as a functional virus receptor.


title: Human metapneumovirus: a newly emerging respiratory pathogen.

text: PURPOSE OF REVIEW The present review focuses on the recent discovery, and clinical and epidemiological features of a virus associated with respiratory tract infections. RECENT FINDINGS In June 2001, researchers in The Netherlands reported the discovery of a virus associated with respiratory tract disease in infants and children. Genetic studies of this newly discovered viral pathogen revealed that it was a paramyxovirus. This virus is the first human pathogen in the Genus Metapneumovirus and was called human metapneumovirus (hMPV). hMPV may be a common respiratory virus. Seroepidemiologic studies in The Netherlands suggested that, by age 5 years, nearly all individuals have been exposed to hMPV. The results of several studies suggest that hMPV may account for about 10% of respiratory tract infections in which a common respiratory virus, such as respiratory syncytial virus, or influenza or parainfluenza viruses, could not be detected. hMPV has been detected in patients with either upper or lower respiratory tract disease, or both. Symptoms associated with hMPV include cough, dyspnea, wheeze, and hypoxia. This newly recognized pathogen has been detected in children and adults. Epidemiological findings suggest that it may circulate worldwide and may have a seasonal distribution. SUMMARY hMPV is a newly emerging respiratory pathogen and may be the cause of a significant proportion of both upper and lower respiratory tract infection in infants, children, and adults.


title: [Detection of severe acute respiratory syndrome probable patients' virus RNA in Hangzhou by using a two loci and a modified nested real-time reverse transcription polymerase chain reaction].

text: OBJECTIVE To detect the RNA of severe acute respiratory syndrome virus (SARS-CoV) by using reverse transcription polymerase chain reaction (RT-PCR) targeted for a two loci and a modified nested real-time RT-PCR as to improving the reliability and sensitivity of tests. METHODS A nested RT-PCR was used for detecting one fragment of SARS-CoV RNA in oropharyngeal swabs from 3 SARS probable patients, 4 SARS suspect patients and other 27 patients with fever in Hangzhou, and the nested RT-PCR product from one SARS probable patient was sequenced. Meanwhile in these 3 SARS probable patients, other three RT-PCR methods, including a hemi-nested RT-PCR targeted for another fragment of SARS-CoV RNA, a real-time RT-PCR and a modified nested real-time RT-PCR, were employed to detect SARS-CoV RNA. RESULTS Two positives were found in the 3 SARS probable patients, and none positive in 4 SARS suspect patients and other 27 patients with fever, using the nested RT-PCR. The sequence of the nested RT-PCR product from one SARS probable patient was identified with the counterpart of SARS-CoV genomes published in public database. The results of the hemi-nested RT-PCR, the real-time RT-PCR and the modified nested real-time RT-PCR in the 3 SARS patients were consistent with the one of the nested RT-PCR. During detecting specimen with low copies of RNA, a weak positive signal was produced after about 35 cycles in the real-time RT-PCR, but a strong positive signal was found only after 10 cycles in the modified nested real-time RT-PCR. CONCLUSION It might improve the reliability of test by employing RT-PCR targeted for two or more fragments in SARS-CoV genome. The modified nested real-time RT-PCR might have higher sensitivity than the routine real-time RT-PCR.


Assessment of neonatal tetanus elimination in Malawi.


title: An Ubiquitin-like Motif in ASK1 Mediates its Association with and Inhibition of the Proteasome.

text: Linear polyubiquitin is processed at LRLRGG sequences by deubiquitinating enzymes to make free monomeric ubiquitin. This LRLRGG ubiquitin-like motif is found in a limited number of mammalian non-ubiquitin proteins, including the MAP3K Apoptosis Signal-Regulating Kinase-1 (ASK1), which activates MAPK signaling pathways. The c-terminus of ASK1 binds to the 19S cap of the proteasome allowing ASK1 to phosphorylate and inhibit proteasomal activity. We investigated whether the ubiquitin-like sequence in the c-terminus of ASK1 mediates its association with and inhibition of the proteasome. To test this we generated ASK1 with substitutions or deletions in this ubiquitin-like domain and examined the activation of cellular signaling and the association of ASK1 with the 19S cap of the proteasome. We show that ASK1 mutants have reduced association with the 19S cap of the proteasome, reduced capacity to inhibit the proteasome, and diminished ability to inhibit TNF-induced NF-κB activation. Mutant forms of ASK1 also had reduced capacity to activate JNK signaling, suggesting that the ubiquitin-like motif in ASK1 is also important for coordinating the balance between JNK and NF-κB signaling. Together these results demonstrate that the ubiquitin-like sequence of ASK1 is important for binding to and inhibition of the proteasome, and for the coordinated activation of cellular NF-κB and JNK signaling.


Article.


title: Serological survey of virus infection among wild house mice (Mus domesticus) in the UK.

text: The serological prevalence of 13 murine viruses was surveyed among 103 wild-caught and 51 captive-bred house mice (Mus domesticus), originating from several trapping locations in northwest England, using blood samples obtained during routine health screening of an established wild mouse colony. A high proportion of recently caught wild mice were seropositive for mouse hepatitis virus (86%), mouse cytomegalovirus (79%), mouse thymic virus (78%), mouse adenovirus (68%), mouse parvovirus (59%) and minute virus of mice (41%). Seroprevalences of lymphocytic choriomeningitis virus (LCMV), orthopoxvirus, reovirus-3 and murid herpesvirus 4 (MuHV-4, also called murine gamma-herpesvirus [MHV-68]) were low (3-13%), and no animals were seropositive to Sendai virus, pneumonia virus or polyomavirus. Seroprevalence in wild-caught animals that had been in captivity for over six months was generally consistent with the range found in recently caught wild animals, while seroprevalence was generally much lower in captive-bred mice despite no attempt to prevent viral spread. A notable exception to this was LCMV, which appeared to have spread efficiently through the captive population (both captive-bred and wild-caught animals). Given the known viral life cycles in laboratory mice, it appears that viral persistence in the host was an important contributing factor in the spread of infection in captivity.


title: Differential effects of human and porcine interleukin 2 on natural killing (NK) activity of newborn piglets and adult pigs lymphocytes.

text: The in vitro effects of human or porcine Interleukin 2 (IL2) on Natural Killing (NK) activity were studied with blood lymphocytes of newborn piglets and of adult pigs. Large volumes of porcine IL2 were prepared by PHA stimulation of irradiated mesenteric lymph node cells and, following purification by gel-filtration chromatography, the apparent molecular weight of IL2 was 15,000 Da. Purified human and porcine IL2 as well as recombinant human IL2 were found to increase markedly NK activity of lymphocytes derived from adult animals. However, although newborn piglets-derived lymphocytes are sensitive to crude porcine IL2 supernatants (and to Interferon alpha as previously shown) their low NK activity is unaffected by purified IL2. These data suggest therefore the existence of differences in the ontogenic development of the lymphokines responsiveness of porcine NK cells.