title: Children hospitalized with severe acute respiratory syndrome-related illness in Toronto.

text: OBJECTIVE An outbreak of severe acute respiratory syndrome (SARS) occurred in the greater Toronto area between February and June 2003. We describe the clinical, laboratory, and epidemiologic features of children who were admitted to the Hospital for Sick Children, Toronto, with a presumptive diagnosis of suspect or probable SARS. METHODS A prospective investigational study protocol was established for the management of children with a presumptive diagnosis of suspect or probable SARS. All were ultimately classified as having probable SARS, suspect SARS, or another cause on the basis of their epidemiologic exposure, clinical and radiologic features, and results of microbiologic investigations. RESULTS Twenty-five children were included; 10 were classified as probable SARS and 5 were classified as suspect SARS, and in 10 another cause was identified. The exposure consisted of direct contact with at least 1 adult probable SARS case in 11 children, travel from a World Health Organization-designated affected area in Asia in 9 children, and presence in a Toronto area hospital in which secondary SARS spread had occurred in 5 children. The predominant clinical manifestations of probable cases were fever, cough, and rhinorrhea. With the exception of 1 teenager, none of the children developed respiratory distress or an oxygen requirement, and all made full recoveries. Mild focal alveolar infiltrates were the predominant chest radiograph abnormality. Lymphopenia; neutropenia; thrombocytopenia; and elevated alanine aminotransferase, aspartate aminotransferase, and creatine kinase were present in some cases. Nasopharyngeal swab specimens were negative for the SARS-associated coronavirus by an in-house reverse transcriptase-polymerase chain reaction in all 25 children. CONCLUSIONS Our results indicate that SARS is a relatively mild and nonspecific respiratory illness in previously healthy young children. The presence of fever in conjunction with a SARS exposure history should prompt one to consider SARS as a possible diagnosis in children irrespective of the presence or absence of respiratory symptoms. Reverse-transcriptase polymerase chain reaction analysis of nasopharyngeal specimens seems to be of little utility for the diagnosis of SARS during the early symptomatic phase of this illness in young children.


title: Illness associated with red tide--Nassau County, Florida, 2007.

text: A "red tide" is a harmful algal bloom that occurs when toxic, microscopic algae in seawater proliferate to a higher-than-normal concentration (i.e., bloom), often discoloring the water red, brown, green, or yellow. Red tides can kill fish, birds, and marine mammals and cause illness in humans. Florida red tide is caused by the dinoflagellate Karenia brevis, which produces toxins called brevetoxins and is most commonly found in the Gulf of Mexico; however, K. brevis blooms also can occur along the Atlantic coast. On September 25, 2007, a cluster of respiratory illnesses was reported to the Nassau County Health Department (NCHD) in northeastern Florida. All of the ill persons were employed at a beach restoration worksite by a dredging company operating at Fernandina Beach; they reported symptoms of eye or respiratory irritation (e.g., coughing, sneezing, sniffling, and throat irritation). NCHD and the Florida Department of Health promptly conducted epidemiologic and environmental investigations and determined the illnesses likely were associated with exposure to a red tide along the Atlantic coast. These actions highlight the importance of rapid investigation of health concerns with potential environmental causes to enable timely notification of the public and prevent further illness.


title: Monoclonal antibodies to the S1 spike and membrane proteins of avian infectious bronchitis coronavirus strain Massachusetts M41.

text: We have established four murine hybridoma cell lines which secrete specific antibody to avian infectious bronchitis virus (IBV) strain Massachusetts M41. Two monoclonal antibodies reacted with the spike protein and two reacted with the membrane protein. The specificity of the monoclonal antibodies for the external structural proteins was detected by immunoprecipitations using radiolabelled virus. The reactions of the monoclonal antibodies showed that one (S1) of the two glycopolypeptides associated with the spike protein has a strain-specific region involved in neutralization and haemagglutination, and the membrane protein has antigenic determinants which are present on the three strains of IBV tested (M41, Beaudette and D41).


title: Development of anti-viral agents using molecular modeling and virtual screening techniques.

text: Computational chemistry has always played a key role in anti-viral drug development. The challenges and the quickly rising public interest when a virus is becoming a threat has significantly influenced computational drug discovery. The most obvious example is anti-AIDS research, where HIV protease and reverse transcriptase have triggered enormous efforts in developing and improving computational methods. Methods applied to anti-viral research include (i) ligand-based approaches that rely on known active compounds to extrapolate biological activity, such as machine learning techniques or classical QSAR, (ii) structure-based methods that rely on an experimentally determined 3D structure of the targets, such as molecular docking or molecular dynamics, and (iii) universal approaches that can be applied in a structure- or ligand-based way, such as 3D QSAR or 3D pharmacophore elucidation. In this review we summarize these molecular modeling approaches as they were applied to fight anti-viral diseases and highlight their importance for anti-viral research. We discuss the role of computational chemistry in the development of small molecules as agents against HIV integrase, HIV-1 protease, HIV-1 reverse transcriptase, the influenza virus M2 channel protein, influenza virus neuraminidase, the SARS coronavirus main proteinase and spike protein, thymidine kinases of herpes viruses, hepatitis c virus proteins and other flaviviruses as well as human rhinovirus coat protein and proteases, and other picornaviridae. We highlight how computational approaches have helped in discovering anti-viral activities of natural products and give an overview on polypharmacology approaches that help to optimize drugs against several viruses or help to optimize the metabolic profile of and anti-viral drug.


title: Laparoscopic assisted low anterior resection for advanced rectal cancer in a kidney transplant recipient: A case report.

text: INTRODUCTION Development of de novo malignancy has become a major cause of late mortality in solid organ transplant recipients. Surgery is currently the most important treatment of choice for transplant patients with resectable CRC. However, conventional open surgery represents a great risk to these high-risk patients. They seem to benefit more from laparoscopic surgery, based on the favorable oncological outcome and remarkable short-term advantages of this approach. PATIENT CONCERNS In this study, we have reported a case of a 50-year-old man who had underwent kidney transplantation for 4 years. He presented with recurrent hematochezia and frequent loose stools for 1 year, and consulted a doctor for recent progressive general malaise and weight loss. DIAGNOSES Colonoscopy revealed a near-circumferential mass at the middle rectum about 8 cm from anal verge. Further biopsy confirmed a diagnosis of adenocarcinoma. Following computed tomography demonstrated peripheral lymph node metastasis, but no signs of distant metastasis. INTERVENTIONS The patient underwent a laparoscopic assisted low anterior resection with total mesorectal excision for rectal cancer. Concomitantly, a loop transverse colostomy was performed to prevent anastomotic leakage. The surgery was completed within 120 min with a blood loss of 100 mL, and immunosuppressive therapy was not stopped perioperatively. Considering the tumor stage of pT3N1M0, the patient also received adjuvant chemotherapy with a regimen of FOLFOX for 8 cycles. OUTCOMES Anastomotic bleeding occurred in this patient about 4 h after surgery, and a control of hemorrhage per anus was performed timely. The following postoperative course was uneventful without any complications, and graft function stayed well. After 4 months of follow-up period, the patient was in a good condition. No evidences of local recurrence and distant metastasis were found. CONCLUSION We have presented a case of successful laparoscopic resection for advanced rectal cancer in a kidney transplant recipient. We believe laparoscopic surgery for CRC in transplant recipients is technically feasible and oncologically safe, which could be a preferred option of surgical procedure in the near future.


title: Prediction of RNA-binding proteins from primary sequence by a support vector machine approach.

text: Elucidation of the interaction of proteins with different molecules is of significance in the understanding of cellular processes. Computational methods have been developed for the prediction of protein-protein interactions. But insufficient attention has been paid to the prediction of protein-RNA interactions, which play central roles in regulating gene expression and certain RNA-mediated enzymatic processes. This work explored the use of a machine learning method, support vector machines (SVM), for the prediction of RNA-binding proteins directly from their primary sequence. Based on the knowledge of known RNA-binding and non-RNA-binding proteins, an SVM system was trained to recognize RNA-binding proteins. A total of 4011 RNA-binding and 9781 non-RNA-binding proteins was used to train and test the SVM classification system, and an independent set of 447 RNA-binding and 4881 non-RNA-binding proteins was used to evaluate the classification accuracy. Testing results using this independent evaluation set show a prediction accuracy of 94.1%, 79.3%, and 94.1% for rRNA-, mRNA-, and tRNA-binding proteins, and 98.7%, 96.5%, and 99.9% for non-rRNA-, non-mRNA-, and non-tRNA-binding proteins, respectively. The SVM classification system was further tested on a small class of snRNA-binding proteins with only 60 available sequences. The prediction accuracy is 40.0% and 99.9% for snRNA-binding and non-snRNA-binding proteins, indicating a need for a sufficient number of proteins to train SVM. The SVM classification systems trained in this work were added to our Web-based protein functional classification software SVMProt, at http://jing.cz3.nus.edu.sg/cgi-bin/svmprot.cgi. Our study suggests the potential of SVM as a useful tool for facilitating the prediction of protein-RNA interactions.


title: Transcription, translation, and cellular localization of three Autographa californica nuclear polyhedrosis virus structural proteins: ODV-E18, ODV-E35, and ODV-EC27.

text: This paper identifies two structural proteins of the occluded derived viral envelope of Autographa californica nuclear polyhedrosis virus (AcMNPV): ODV-E18 and ODV-E35. In addition, we identify a protein, ODV-EC27, that is incorporated into the capsid of occluded virus, which is not detected in budded virus. The genes for these proteins reside within the IE0 intron. The intron was sequenced, and five open reading frames (ORF) were identified. ORF 3 (genomic ORF 143) codes for the ODV envelope protein, ODV-E18. ORF 4 (genomic ORF 144) codes for ODV-EC27, and Western blot analyses locate this protein to both the ODV capsid and envelope. Transcripts for both ODV-E18 and ODV-EC27 initiate from conserved TAAG motifs, and transcripts are detected from 16 through 72 hr p.i. Antiserum to ODV-E18 recognizes a band of 18 kDa on Western blots of extracts from infected cells and bands of 18 and 35 kDa on Western blots of proteins from purified ODV envelope. N-terminal amino acid sequencing reveals that both ODV-E18 and ODV-E35 contain the same N-terminus. Antiserum to ODV-EC27 recognizes a protein of 27 kDa on Western blots of extracts from infected cells and bands of 27 and 35 kDa on Western blots of proteins from purified ODV. Using immunogold labeling techniques, ODV-E18 and/or ODV-E35 are detected in viral induced intranuclear microvesicles and are not detected in the plasma membrane, cytoplasmic membranes, or the nuclear envelope. Immunogold labeling using antisera to ODV-EC27 detects this protein on both the ODV envelope and capsid.


title: CDC's Early Response to a Novel Viral Disease, Middle East Respiratory Syndrome Coronavirus (MERS-CoV), September 2012-May 2014.

text: The first ever case of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) was reported in September 2012. This report describes the approaches taken by CDC, in collaboration with the World Health Organization (WHO) and other partners, to respond to this novel virus, and outlines the agency responses prior to the first case appearing in the United States in May 2014. During this time, CDC's response integrated multiple disciplines and was divided into three distinct phases: before, during, and after the initial activation of its Emergency Operations Center. CDC's response to MERS-CoV required a large effort, deploying at least 353 staff members who worked in the areas of surveillance, laboratory capacity, infection control guidance, and travelers' health. This response built on CDC's experience with previous outbreaks of other pathogens and provided useful lessons for future emerging threats.


title: Acquiring Pharmaceutical Industry Assets in the UK: 1 + 1 = 1?

text: The recent AstraZeneca takeover bid from Pfizer puts pharmaceutical R&D once again on the public agenda. Three pertinent questions are (a) what can be expected from this acquisition, (b) what are the implications for the UK economy and science base, and (c) whether such a deal should go ahead. Although the key driver behind this acquisition would be an improvement in company performance and shareholder value, past evidence suggests that mergers and acquisitions (M&A) of large pharmaceutical companies imply a neutral net effect on productivity, if not a decline, with employment decreasing and R&D spend following a similar trend. Similarities between the two companies include dropping sales; however, relative to its size, AstraZeneca has a more promising R&D pipeline, especially in therapeutic areas where Pfizer's strength is currently limited (e.g. oncology). Ensuring a portfolio diversification would make Pfizer's takeover proposal a knight's one, but history points towards a knave-like behavior.


title: Serological responses in patients with severe acute respiratory syndrome coronavirus infection and cross-reactivity with human coronaviruses 229E, OC43, and NL63.

text: The serological response profile of severe acute respiratory syndrome (SARS) coronavirus (CoV) infection was defined by neutralization tests and subclass-specific immunofluorescent (IF) tests using serial sera from 20 patients. SARS CoV total immunoglobulin (Ig) (IgG, IgA, and IgM [IgGAM]) was the first antibody to be detectable. There was no difference in time to seroconversion between the patients who survived (n = 14) and those who died (n = 6). Although SARS CoV IgM was still detectable by IF tests with 8 of 11 patients at 7 months postinfection, the geometric mean titers dropped from 282 at 1 month postinfection to 19 at 7 months (P = 0.001). In contrast, neutralizing antibody and SARS CoV IgGAM and IgG antibody titers remained stable over this period. The SARS CoV antibody response was sometimes associated with an increase in preexisting IF IgG antibody titers for human coronaviruses OC43, 229E, and NL63. There was no change in IF IgG titer for virus capsid antigen from the herpesvirus that was used as an unrelated control, Epstein-Barr virus. In contrast, patients who had OC43 infections, and probably also 229E infections, without prior exposure to SARS CoV had increases of antibodies specific for the infecting virus but not for SARS CoV. There is a need for awareness of cross-reactive antibody responses between coronaviruses when interpreting IF serology.


Host factors in positive-strand RNA virus genome replication.


title: The importance of pathogenic organisms in sewage and sewage sludge.

text: Deficient sanitation poses a serious threat to human and animal health, involving complex relationships between environments, animals, refuse, food, pathogens, parasites, and man. However, by sanitizing and stabilizing the organic matter of sewage sludge, agriculture can utilize it to maintain soil, water, and air quality. As ingredients in soil amendments, such bioresidues are a source of nutrients for plants. Stabilization and sanitation of sewage sludge safely couple its recycling and disposal. This coupling becomes increasingly important as economic and environmental constraints make strategies for waste disposal more difficult to apply. The occurrence of viruses, bacteria, yeasts, fungi, and zooparasites in sewage sludge is reviewed in this article, and consequential epidemiologic concerns that arise from sewage sludge recycling is also addressed.


title: Detection of antibodies to human coronaviruses 229E and OC43 in the sera of multiple sclerosis patients and normal subjects.

text: Sera collected from 90 multiple sclerosis patients and 148 age-matched normal subjects were examined for the presence of antibodies against human coronaviruses (HCV) 229E and OC43 by enzyme immunoassay (EIA). The results demonstrated no significant difference between the MS patients and the normal subjects in their antibody titer to HCV 229E and HCV OC43. Further analysis of these 238 sera indicated that a stronger EIA reaction was generally observed against HCV OC43 (mean EIA value at an optical density of 492 nm = 0.896) than against HCV 229E (mean EIA value at an optical density of 492 nm = 0.346).


title: Assessing public health capabilities during emergency preparedness tabletop exercises: reliability and validity of a measurement tool.

text: OBJECTIVES Improving the ability of local public health agencies to respond to large-scale emergencies is an ongoing challenge. Tabletop exercises can provide an opportunity for individuals and groups to practice coordination of emergency response and evaluate performance. The purpose of this study was to develop a valid and reliable self-assessment performance measurement tool for tabletop exercise participants. METHODS The study population comprised 179 public officials who attended three tabletop exercises in Massachusetts and Maine between September 2005 and November 2006. A 42-item questionnaire was developed to assess five public health functional capabilities: (1) leadership and management, (2) mass casualty care, (3) communication, (4) disease control and prevention, and (5) surveillance and epidemiology. Analyses were undertaken to examine internal consistency, associations among scales, the empirical structure of the items, and inter-rater agreement. RESULTS Thirty-seven questions were retained in the final questionnaire and grouped according to the original five domains. Alpha coefficients were 0.81 or higher for all scales. The five-factor solution from the principal components analysis accounted for 60% of the total variance, and the factor structure was consistent with the five domains of the original conceptual model. Inter-rater agreement ranged from good to excellent. CONCLUSIONS The resulting 37-item performance measurement tool was found to reliably measure public health functional capabilities in a tabletop exercise setting, with preliminary evidence of a factor structure consistent with the original conceptualization and of criterion-related validity.


title: Molecular targets for diagnostics and therapeutics of severe acute respiratory syndrome (SARS-CoV).

text: PURPOSE The large number of deaths in a short period of time due to the spread of severe acute respiratory syndrome (SARS) infection led to the unparalleled collaborative efforts world wide to determine and characterize the new coronavirus (SARS-CoV). The full genome sequence was determined within weeks of the first outbreak by the Canadian group with international collaboration. As per the World Health Organization (WHO), the continual lack of a rapid laboratory test to aid the early diagnosis of suspected cases of SARS makes this area a priority for future research. To prevent deaths in the future, early diagnosis and therapy of this infectious disease is of paramount importance. METHODS This review describes the specific molecular targets for diagnostics and therapeutics of viral infection. RESULTS The three major diagnostic methods available for SARS includes viral RNA detection by reverse transcription polymerase chain reaction (RT-PCR), virus induced antibodies by immunofluorescence assay (IFA) or by enzyme linked immunosorbant assay (ELISA) of nucleocapsid protein (NP). The spike glycoprotein of SARS-CoV is the major inducer of neutralizing antibodies. The receptor binding domain (RBD) in the S1 region of the spike glycoprotein contains multiple conformational epitopes that induces highly potent neutralizing antibodies. The genetically engineered attenuated form of the virus or viral vector vaccine encoding for the SARS-CoV spike glycoprotein has been shown to elicit protective immunity in vaccinated animals. CONCLUSION NP is the preferred target for routine detection of SARS-CoV infection by ELISA which is an economical method compared to other methods. The RBD of the spike glycoprotein is both a functional domain for cell receptor binding and also a major neutralizing determinant of SARS-CoV. The progress in evaluating a therapeutic or vaccine would depend on the avail ability of clinically relevant animal model.


title: An sveir model for assessing potential impact of an imperfect anti-sars vaccine.

text: The control of severe acute respiratory syndrome (SARS), a fatal contagious viral disease that spread to over 32 countries in 2003, was based on quarantine of latently infected individuals and isolation of individuals with clinical symptoms of SARS. Owing to the recent ongoing clinical trials of some candidate anti-SARS vaccines, this study aims to assess, via mathematical modelling, the potential impact of a SARS vaccine, assumed to be imperfect, in curtailing future outbreaks. A relatively simple deterministic model is designed for this purpose. It is shown, using Lyapunov function theory and the theory of compound matrices, that the dynamics of the model are determined by a certain threshold quantity known as the control reproduction number (R(v)). If R(v) =/< 1, the disease will be eliminated from the community; whereas an epidemic occurs if R(v) > 1. This study further shows that an imperfect SARS vaccine with infection-blocking efficacy is always beneficial in reducing disease spread within the community, although its overall impact increases with increasing efficacy and coverage. In particular, it is shown that the fraction of individuals vaccinated at steady-state and vaccine efficacy play equal roles in reducing disease burden, and the vaccine must have efficacy of at least 75% to lead to effective control of SARS (assuming R(0) = 4). Numerical simulations are used to explore the severity of outbreaks when R(v) > 1.


title: Severe acute respiratory infections surveillance in Romania - is it any gain to public health?

text: UNLABELLED This article aims to present the operation and usefulness of severe acute respiratory infection (SARI) surveillance in Romania. Material and methods : The information obtained from surveillance data analysis using Epilnfo program revealed a greater severity of influenza in the second post-pandemic season (2011/2012) compared to the previous one (2010/2011). Detected risk factors were evidence-based arguments for recommending preventive measures in certain population groups. CONCLUSIONS This surveillance system has proven useful, being able to provide information on the severity of influenza and other etiologies involved in severe acute respiratory infections.


title: Renal flagellate infections in reptiles: 29 cases.

text: Renal infection with flagellated protozoa was retrospectively evaluated in 29 reptiles, including 12 turtles, 7 tortoises, and 6 chameleons; overall, 20 species of reptiles were represented. Most cases presented with nonspecific clinical signs or a combination of several concurrent diseases. Nineteen of 29 reptiles had tubulointerstitial nephritis associated with flagellates, and this lesion was considered contributory to death in 15 cases, although concurrent diseases were frequent. Infection was invasive into the renal interstitium in three reptiles due to tubular rupture and in one chameleon also spread to adjacent tissues, coelomic cavity, and blood vessels due to renal rupture. Cytologic or ultrastructural evaluation of trophozoites in two cases was consistent with diplomonad flagellates. Renal disease was often complicated with soft-tissue mineralization and/or gout. Gastrointestinal and cloacal infection with flagellates and inflammation were frequent in reptiles in which the digestive tract was available for histopathologic examination, and this supports the possibility of infections ascending the urinary tract from the cloaca. Renal disease associated with flagellate protozoa is rare in vertebrates but appears to be relevant in reptiles, particularly chelonians and chameleons.


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

text: A new protein of the feline infectious peritonitis virus (FIPV) was discovered in lysates of infected cells. Expression of the gene encoding open reading frame (ORF) 6b of FIPV in recombinant vaccinia virus infected cells was used to identify it as the 6b protein. 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 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.


Impact in the Fight Against COVID-19.


COVID-19 Pandemic: Impact on Healthcare in Missouri.


title: Principia aetiologica: taking causality beyond Koch's postulates.

text: There is no single accepted method to establish a causal relationship between an infective agent and its corresponding infectious disease. Different biomedical disciplines use a patchwork of distinct but overlapping approaches. To a greater or lesser extent these are based on criteria known as the Koch-Henle postulates, or 'Koch's postulates' for short. Deficiencies in Koch's postulates were recognized by their principal author shortly after their formulation. Now, over a century later, a more rigorous method to test causality has still to be finalized. One contender is a method that uses molecular methods to establish a causal relationship ('molecular Koch's postulates'). Recognizing the wider range of contemporary approaches used to build an argument for a causal relationship, the use of a more inclusive approach to establish proof of causality is proposed. This method uses an argument built from a series of assertions. Assertion 1: congruence or reproducible correlation of a taxonomically defined life form with the clinico-pathological and epidemiological features of infection. Assertion 2: consistency of the demonstrable biological response in the subject to an encounter with the prospective infective agent. Assertion 3: progressive or cumulative dissonance as an explanation for pathophysiological processes at every known level of biological organization in the subject. Assertion 4: curtailment of that pathophysiological process on the deliberate introduction of a specified biomedical intervention. Evidence to implicate the candidate biological entity as an initiator of or primer for cumulative dissonance places it in a subcategory of micro-organisms to be known as 'priobes'. A priobe is the sufficient and necessary antecedent cause of a pathophysiological process evident as an infectious disease.


Enteric infections in man and animals: standardization of immunological procedures. Proceedings of a symposium: International Association of Biological Standardization co-sponsored by the World Health Organization. Dublin, Ireland, September 6-8, 1982.


title: Translational control of viral gene expression in eukaryotes.

text: As obligate intracellular parasites, viruses rely exclusively on the translational machinery of the host cell for the synthesis of viral proteins. This relationship has imposed numerous challenges on both the infecting virus and the host cell. Importantly, viruses must compete with the endogenous transcripts of the host cell for the translation of viral mRNA. Eukaryotic viruses have thus evolved diverse mechanisms to ensure translational efficiency of viral mRNA above and beyond that of cellular mRNA. Mechanisms that facilitate the efficient and selective translation of viral mRNA may be inherent in the structure of the viral nucleic acid itself and can involve the recruitment and/or modification of specific host factors. These processes serve to redirect the translation apparatus to favor viral transcripts, and they often come at the expense of the host cell. Accordingly, eukaryotic cells have developed antiviral countermeasures to target the translational machinery and disrupt protein synthesis during the course of virus infection. Not to be outdone, many viruses have answered these countermeasures with their own mechanisms to disrupt cellular antiviral pathways, thereby ensuring the uncompromised translation of virion proteins. Here we review the varied and complex translational programs employed by eukaryotic viruses. We discuss how these translational strategies have been incorporated into the virus life cycle and examine how such programming contributes to the pathogenesis of the host cell.


title: -Entrance of PCR for studying murine coronaviruses-.

text: This paper describes a number of applications of the Polymerase Chain Reaction (PCR) in the study of enterotropic murine coronaviruses [mouse hepatitis virus (MHV)]. A diagnostic PCR was developed which detected all of 11 different MHV strains. This fast and reliable method was also able to differentiate MHV from other non-murine coronaviruses. On the basis of this assay a quantitative PCR was designed using a mutant template containing a point mutation which competed for the PCR primers. The amplification and cloning of the structural protein genes of enterotropic MHV strains it plasmid vectors for subsequent sequencing is described. In addition an RT PCR was developed which was able to selectivity detect artificially generated recombinant coronavirus.


title: [Video-laryngoscope (McGRATH MAC): a clinical assessment of its performance in consecutive 100 patients].

text: BACKGROUND The McGRATH MAC Video laryngoscope (Covidien Japan) is a relatively new device for tracheal intubation, which provides an excellent glottis view. We here report the clinical experience of the McGRATH in 100 patients who received general anesthesia. METHODS Tracheal intubation with the McGRATH was performed in 100 patients requiring oro/naso tracheal intubation. The view of glottic opening was scored according to the classification of Cormack-Lehane. The time required to intubate and the number of intubation attempts were recorded. RESULTS Cormack-Lehane classification 1 or 2 was obtained in 99 of 100 patients, respectively, and successful intubation was achieved in all patients. The mean +/- SD time to successful intubation was 40 +/- 19 seconds. CONCLUSIONS The McGRATH was easily handled not only by experienced anesthetists but also by novice personnel. The McGRATH seems to be a useful device in routine and difficult airway management.


title: Profile of antibodies to the nucleocapsid protein of the severe acute respiratory syndrome (SARS)-associated coronavirus in probable SARS patients.

text: Profiles of antibodies to the nucleocapsid protein of the severe acute respiratory syndrome (SARS)-associated coronavirus in 445 probable SARS patients and 3,749 healthy people or non-SARS patients were analyzed by antigen-capturing enzyme-linked immunosorbent assay. Antinucleocapsid antibodies were elucidated in 17.5% of the probable SARS patients 1 to 7 days after the onset of symptoms and in 80% of the patients 8 to 14 days after the onset. About 90% of the probable SARS patients were positive 15 or more days after illness. Antibody titers increased up to 70 days, and high antibody titers were maintained at least for another 3 months. Of the healthy people and non-SARS patients, only seven (0.187%) were weakly positive.


title: Hematological findings in SARS patients and possible mechanisms (review).

text: Severe acute respiratory syndrome (SARS) is a new human infectious disease. The causative agent of SARS is a novel coronavirus (SARS-CoV). This report summarizes the hematological findings in SARS patients and proposes the possible mechanisms of SARS-CoV related abnormal hematopoiesis. Hematological changes in patients with SARS are common and include lymphopenia, thrombocytopenia and occasionally leukopenia. A significant decrease was also observed in peripheral CD4+ and CD8+ T lymphocyte subsets and it was related to onset of SARS. A number of potential mechanisms may be involved. The development of auto-immune antibodies or immune complexes triggered by viral infection may play a major role in inducing lymphopenia and thrombocytopenia. Moreover, SARS-CoV may also directly infect hematopoietic stem/progenitor cells via CD13 or CD66a inducing their growth inhibition and apoptosis. The receptor for group I and III CoV is aminopeptidase N (CD13). CD13 has been identified in human bone marrow CD34+ cells, platelets, megakaryocytes, myeloid cells, and erythroid cells, but not in lymphocytes. The common receptor for group II CoV is CEACAM1a (CD66a). CD66a is an adhesion molecule expressed on bone marrow CD34+ cells, platelets, granulocytes and activated lymphocytes. In addition, glucocorticoids could induce lymphopenia and the use of steroids may account for the decrease of lymphocytes in some SARS patients. The increased consumption of platelets and/or the decreased production of platelets in the damaged lungs are a potential alternative but often overlooked mechanism that can contribute to thrombocytopenia in severe critical pulmonary conditions.


title: Influenza A virus among the hospitalized young children with acute respiratory infection. Is influenza A co infected with respiratory syncytial virus?

text: BACKGROUND Both influenza A virus (IAV) and respiratory syncytial virus (RSV) cause acute respiratory infection (ARI) in infants and young children. This study was conducted to determine Influenza A virus and its co infection with RSV among the hospitalized children with ARI. METHODS A total of 153 throat samples of the hospitalized young children aged between below one year and 5 years with the clinical signs of ARI were collected from the different hospitals in Khuzestan from June 2009 to April 2010. The samples were tested for Influenza A viruses by real time PCR. Positive IAV samples were tested for influenza A sub type H1N1 and for RSV by the nested PCR. RESULTS In this study, from the total 153 samples, 35 samples (22.9%) including 15 (42.8%) females and 20 (57.2%) males were positive for influenza A viruses. From the 35 positive samples for IAV, 14 were positive for swine H1N1 subtype. All the positive samples for influenza showed negative for RSV infection which revealed no coinfection with RSV. The prevalence of influenza A among age/sex groups was not significant. CONCLUSION Influenza A is a prevalent viral agent isolated from young children with ARI. Influenza A subtype H1N1 was accounted for the 40 percent all laboratory-proven diagnoses of influenza in 2009. No evidence of coinfection of influenza A and RSV has been observed in the present study.


Role of host age and genotype in murine enterotropic coronavirus infection.


title: Evidence for recombination between feline panleukopenia virus and canine parvovirus type 2.

text: Canine parvovirus type 2 (CPV) is a virulent pathogen that emerged in the late 1970s, probably originating from feline panleukopenia virus (FPLV) or a closely related carnivore parvovirus belonging to the feline parvovirus (FPV) subspecies. In contrast to FPLV, CPV has evolved rapidly since its emergence. The original antigenic type of CPV disappeared more than two decades ago and several new antigenic as well as genetic CPV variants have appeared and spread in the field. Both high mutation rate and positive selection of mutations in the capsid gene appear to be the driving force for such rapid evolution. In addition, genetic recombination has been assessed as a factor in parvovirus evolution. Recently, we provided the first evidence of inter-antigenic type recombination of CPV in nature. Here, an inter-FPV subspecies recombinant was revealed by analyzing the genetic data deposited in databases with several recombination detection programs, and by phylogeny. FPLV strain XJ-1, submitted by Su et al., Harbin, China in 2007 (GenBank accession no. EF988660), was most likely generated by recombination between CPV and FPLV. Its genome was generally composed of the NS1 gene of CPV origin and the VP1 gene of FPLV origin. This is the first demonstration of recombination between different FPV subspecies in nature. Consequently, recombination should be considered as an element in the generation and evolution of parvoviruses of the FPV subspecies.


title: Evaluation of the role of heterogeneous nuclear ribonucleoprotein A1 as a host factor in murine coronavirus discontinuous transcription and genome replication.

text: Viruses with RNA genomes often capture and redirect host cell components to assist in mechanisms particular to RNA-dependent RNA synthesis. The nidoviruses are an order of positive-stranded RNA viruses, comprising coronaviruses and arteriviruses, that employ a unique strategy of discontinuous transcription, producing a series of subgenomic mRNAs linking a 5' leader to distal portions of the genome. For the prototype coronavirus mouse hepatitis virus (MHV), heterogeneous nuclear ribonucleoprotein (hnRNP) A1 has been shown to be able to bind in vitro to the negative strand of the intergenic sequence, a cis-acting element found in the leader RNA and preceding each downstream ORF in the genome. hnRNP A1 thus has been proposed as a host factor in MHV transcription. To test this hypothesis genetically, we initially constructed MHV mutants with a very high-affinity hnRNP A1 binding site inserted in place of, or adjacent to, an intergenic sequence in the MHV genome. This inserted hnRNP A1 binding site was not able to functionally replace, or enhance transcription from, the intergenic sequence. This finding led us to test more directly the role of hnRNP A1 by analysis of MHV replication and RNA synthesis in a murine cell line that does not express this protein. The cellular absence of hnRNP A1 had no detectable effect on the production of infectious virus, the synthesis of genomic RNA, or the quantity or quality of subgenomic mRNAs. These results strongly suggest that hnRNP A1 is not a required host factor for MHV discontinuous transcription or genome replication.


title: Cell culture-grown putative bovine respiratory torovirus identified as a coronavirus.

text: A putative bovine respiratory torovirus (BRTV) was propagated in bovine fetal diploid lung and human colonic tumour cells, and fringed pleomorphic particles were detected in the culture supernatants by electron microscopy. Antisera directed against a bovine (Breda strain) and equine (Berne strain) torovirus failed to react with BRTV-infected cells in immunofluorescence assays and did not neutralise BRTV. No toroviral RNA was found in the supernatants of infected cells by means of a reverse transcriptase-polymerase chain reaction with torovirus-specific primers. On the other hand, bovine coronavirus-specific antisera and monoclonal antibodies did neutralise the cytopathic effects, and coronaviral antigen was detected in the cultures by immunofluorescence. Furthermore, bovine coronavirus RNA was detected in the supernatants of BRTV-infected cells after nucleic acid amplification. It is concluded that the cytopathic BRTV isolate is a coronavirus.


title: Quantitative analysis of cytokine mRNA expression and protozoan DNA load in Theileria parva-infected cattle.

text: Theileria parva (T. parva) causes a highly serious bovine disease called East Coast fever (ECF), which is characterized by pyrexia, dyspnea and cachexia and is of great economic importance in African countries. We hypothesize that the clinical symptoms of ECF could be explained by a cytokine dysregulation. In this study, we investigated the relationship between T. parva DNA load and expression levels of cytokine mRNAs in leukocytes from experimentally infected calves by quantitative PCR. The p104 gene, which encodes the T. parva 104 kDa microneme-rhoptry protein, was detected in cattle blood from day 10 after T. parva-infected tick infestation, and the protozoan DNA load was increased together with severity of disease. The mRNA expressions of pro-inflammatory cytokines, such as interleukin (IL)-1beta and IL-6, were up-regulated with protozoan DNA load increasing. In addition, the level of a type-2 cytokine (IL-10) transcript was also increased during the acute phase. In contrast, the down-regulation or no detectable levels of the expression of type-1 cytokines, such as IL-2 and interferon (IFN)-gamma were observed in T. parva-infected animals. Thus, our observations indicated that high protozoan load and resulting intense inflammatory responses might be involved in the severity of clinical signs observed in T. parva-infection.


title: Chemical modification: the key to clinical application of RNA interference?

text: RNA interference provides a potent and specific method for controlling gene expression in human cells. To translate this potential into a broad new family of therapeutics, it is necessary to optimize the efficacy of the RNA-based drugs. As discussed in this Review, it might be possible to achieve this optimization using chemical modifications that improve their in vivo stability, cellular delivery, biodistribution, pharmacokinetics, potency, and specificity.


title: Polio control after certification: major issues outstanding.

text: Now that the global eradication of wild poliovirus is almost within sight, planning for the post-certification era is becoming a priority issue. It is agreed that a stockpile of appropriate polio vaccines will need to be established, and a surveillance and response capacity will need to be maintained, in order to protect the world against any possible future outbreaks attributable either to the persistence of wild poliovirus or vaccine-derived polioviruses (VDPVs) or to the unintentional or intentional release of poliovirus from a laboratory or vaccine store. Although it has been suggested that the stockpile should consist of monovalent oral poliovirus vaccine (mOPV), many questions remain concerning its nature, financing, management, and use--in particular, because of uncertainties over future national vaccination policies, and over the availability of different vaccines, after the certification of wild poliovirus eradication. There are further uncertainties concerning the possible role and efficacy of inactivated poliovirus vaccine (IPV) used either routinely or in outbreak control in low-hygiene settings, the potential for rapid geographical spread of polioviruses should an outbreak occur after certification, and the risks inherent in introducing additional oral polio vaccine (OPV) viruses into populations in which the vaccine coverage and prevalence of immunity have declined, and which may thus favour the spread of VDPVs. Given these important gaps in knowledge, no country should discontinue polio vaccination until a coordinated policy for the post-certification era has been developed and the recommended measures have been put in place.


Equine viral arteritis.


Exploiting DNA immunization to generate polyclonal antisera to coronavirus replicase proteins.


title: Position statement. Part one: Immune function and exercise.

text: An ever-growing volume of peer-reviewed publications speaks to the recent and rapid growth in both scope and understanding of exercise immunology. Indeed, more than 95% of all peer-reviewed publications in exercise immunology (currently >2, 200 publications using search terms "exercise" and "immune") have been published since the formation of the International Society of Exercise and Immunology (ISEI) in 1989 (ISI Web of Knowledge). We recognise the epidemiological distinction between the generic term "physical activity" and the specific category of "exercise", which implies activity for a specific purpose such as improvement of physical condition or competition. Extreme physical activity of any type may have implications for the immune system. However, because of its emotive component, exercise is likely to have a larger effect, and to date the great majority of our knowledge on this subject comes from exercise studies.


title: [Severe acute respiratory syndrome (SARS)].

text: Severe Acute Respiratory Syndrome (SARS) is a disease caused by a corona-virus. These viruses are long known to be human pathogens but this form is new and more virulent. It emerged in March this year in China and seemed to be apt to sweep over the world as a kind of Spanish Flu. Forunately, this didn't happen and at the moment the emerging pandemic seems to be contained. This offers hope in case a more virulent pandemic would suddenly strike in the next future. A world prepared is a difficult prey, even for a surprise virus.


title: Dynamic changes in clinical features and cytokine/chemokine responses in SARS patients treated with interferon alfacon-1 plus corticosteroids.

text: Severe acute respiratory syndrome (SARS), caused by a novel coronavirus, emerged in early 2003 as a major international health crisis. We report on serum cytokine levels, viral load and clinical parameters over the course of the disease in a cohort of nine adult SARS patients treated with steroids and interferon alfacon-1 at North York General Hospital in Toronto, Ontario. Considerable variation among SARS patients with respect to circulating viral load and patterns of SARS-CoV-evoked cytokine responses was recorded. No single cytokine profile was observed in all patients, yet serum concentrations of interferon (IFN)-gamma, interleukin (IL)-10, CXCL10, CCL5 and CXCL8 were found to be elevated above normal levels during the course of the disease in all patients. Expression levels for IL-10, IFN-gamma and CXCL10 consistently peaked within 4 days of peak viral load. IL-12p70, IL-4 and tumour necrosis factor-alpha concentrations were consistently highest within 5 days of peak viral load. These results suggest that elevated levels of inflammatory cytokines are sensitive correlates of disease severity, including lung abnormalities and viral load in serum, and may provide a tool for monitoring disease progression in affected individuals.


title: Global Health Security: Building Capacities for Early Event Detection, Epidemiologic Workforce, and Laboratory Response.

text: The Global Health Security Agenda (GHSA) was launched in February 2014 to bring countries with limited capacity into compliance with the International Health Regulations (IHR) (2005). Recent international public health events, such as the appearance of Middle Eastern respiratory syndrome coronavirus and the reappearance of Ebola in West Africa, have highlighted the importance of early detection of disease events and the interconnectedness of countries. Surveillance systems that allow early detection and recognition of signal events, a public health infrastructure that allows rapid notification and information sharing within countries and across borders, a trained epidemiologic workforce, and a laboratory network that can respond appropriately and rapidly are emerging as critical components of an early warning and response system. This article focuses on 3 aspects of the GHSA that will lead to improved capacities for the detection and response to outbreaks as required by the IHR: (1) early detection and reporting of events, (2) laboratory capacity, and (3) a trained epidemiologic workforce.


title: Evaluation of a national pharmacy-based syndromic surveillance system.

text: Background Traditional public health surveillance provides accurate information but is typically not timely. New early warning systems leveraging timely electronic data are emerging, but the public health value of such systems is still largely unknown. Objective To assess the timeliness and accuracy of pharmacy sales data for both respiratory and gastrointestinal infections and to determine its utility in supporting the surveillance of gastrointestinal illness. Methods To assess timeliness, a prospective and retrospective analysis of data feeds was used to compare the chronological characteristics of each data stream. To assess accuracy, Ontario antiviral prescriptions were compared to confirmed cases of influenza and cases of influenza-like-illness (ILI) from August 2009 to January 2015 and Nova Scotia sales of respiratory over-the-counter products (OTC) were compared to laboratory reports of respiratory pathogen detections from January 2014 to March 2015. Enteric outbreak data (2011-2014) from Nova Scotia were compared to sales of gastrointestinal products for the same time period. To assess utility, pharmacy sales of gastrointestinal products were monitored across Canada to detect unusual increases and reports were disseminated to the provinces and territories once a week between December 2014 and March 2015 and then a follow-up evaluation survey of stakeholders was conducted. Results Ontario prescriptions of antivirals between 2009 and 2015 correlated closely with the onset dates and magnitude of confirmed influenza cases. Nova Scotia sales of respiratory OTC products correlated with increases in non-influenza respiratory pathogens in the community. There were no definitive correlations identified between the occurrence of enteric outbreaks and the sales of gastrointestinal OTCs in Nova Scotia. Evaluation of national monitoring showed no significant increases in sales of gastrointestinal products that could be linked to outbreaks that included more than one province or territory. Conclusion Monitoring of pharmacy-based drug prescriptions and OTC sales can provide a timely and accurate complement to traditional respiratory public health surveillance activities but initial evaluation did not show that tracking gastrointestinal-related OTCs were of value in identifying an enteric disease outbreak in more than one province or territory during the study period.


Endoscopic endonasal resection of the odontoid peg--case report and literature review.


title: Clinical usefulness of endoscopic ultrasonography with or without fine needle aspiration in the diagnosis and staging of pancreatic carcinoma.

text: INTRODUCTION The aims of this study was to show the accuracy and clinical usefulness of endoscopic ultrasonography (EUS) with EUS-guided fine needle aspiration (FNA) in the diagnosis and staging of pancreatic cancer not obvious in computed tomographic (CT) scan abdomen imaging. MATERIALS AND METHODS Five male patients were evaluated; 4 presented with obstructive jaundice and 1 had unexplained loss of weight. The mean age was 66 years (range, 40 to 77). All had CT scan abdomen imaging which did not show any obvious pancreatic tumour. EUS with FNA was done for all cases when indicated. Surgical findings, if any, were obtained and compared to EUS findings. RESULTS EUS easily detected the pancreatic tumour in all 5 cases. The tumour sizes detected ranged from 27 to 40 mm in diameter. These corresponded fairly accurately with that of surgical findings for all 3 who had surgery. EUS reported 3 cases with pathological lymph node involvement. All 5 cases were confirmed by FNA or surgery. EUS was also accurate in 4 cases, which reported the absence of portal vein or superior mesenteric vein invasion. Surgical documentation could not verify the fifth case. There were no complications at all from the EUS with/without FNA. CONCLUSION This case series showed that EUS with/without FNA appears to be useful and safe in diagnosing and staging pancreatic head tumours not detectable by CT scanning.


title: The aetiology and treatment of oral halitosis: an update.

text: Halitosis refers to the condition of offensive mouth odour. More than 90% of cases of halitosis originate from the oral cavity. The implicated bacteria (Fusobacterium nucleatum, Prevotella intermedia, and Tannerella forsythensis) are located in stagnant areas in the oral cavity, such as the dorsal surface of tongue, periodontal pockets, and interproximal areas. These bacteria proteolyse the amino acids releasing volatile sulphur compounds. The management of halitosis involves determining and eliminating the causes, which includes identifying any contributory factors, because certain medical conditions are also associated with characteristic smells. Professional advice should be given on oral hygiene and diet, and treatments should include dental scaling, and root planing of the associated periodontal pockets to reduce the bacterial loading. In addition to the normal oral hygiene practice, tongue cleaning and use of mouthwash are advocated. This paper discusses the common aetiological factors, classification of oral halitosis, and its treatment.


Unilateral mandibular hyperostosis in a cat.


title: A multicenter study of grepafloxacin and clarithromycin in the treatment of patients with community-acquired pneumonia.

text: STUDY OBJECTIVES To compare the efficacies of 10-day regimens of grepafloxacin (GFX) (Raxar or Vaxar; Glaxo Wellcome; Greenford, UK), 600 qd, and clarithromycin (CLA) (Klacid, Biaxin, or Klaracid; Abbott Laboratories; Chicago, IL), 500 mg bid, in patients with community-acquired pneumonia (CAP), on the basis of clinical response, including radiographic evidence, and bacteriologic efficacy. DESIGN Phase IIIb, double-blind, double-dummy, randomized, prospective, parallel-group, comparative study conducted at 58 centers in 11 countries. PATIENTS AND SETTING Adult patients with signs and symptoms of CAP that was confirmed by radiographic evidence and who did not require parenteral therapy were included in the study. ASSESSMENTS Patients were assessed before treatment, during treatment, after treatment, and at follow-up (28 to 35 days after treatment completion). Clinical response was evaluated. Blood and sputum samples were cultured for bacterial pathogens, and serology testing was performed to detect atypical pneumonia. RESULTS A total of 504 patients were enrolled into the trial: 251 were randomly assigned to receive GFX and 253 to receive CLA. In patients able to be clinically evaluated, clinical success rates at follow-up were 147 of 163 patients (90%) in the GFX group and 148 of 167 patients (89%) in the CLA group (95% confidence interval, -6% to 9%). Pretreatment pathogens were confirmed in 131 of 504 patients (26%), either by culture or serology testing, the primary pathogens being Streptococcus pneumoniae (22%), Haemophilus influenzae (17%), Mycoplasma pneumoniae (25%), and Chlamydia pneumoniae (11%). For patients able to be evaluated who had typical pathogens, bacteriologic success was achieved in 92% of the patients in each treatment group. For patients able to be evaluated who had atypical pathogens, 18 of 18 patients (100%) in the GFX and 23 of 26 patients (88%) in the CLA group had a successful clinical outcome. There were similar rates of adverse events in each group, resulting in </= 7% withdrawal from treatment; gastrointestinal events were the most common. CONCLUSIONS GFX, 600 mg qd, was equivalent to CLA, 500 mg bid, in treating adult patients with CAP. Both treatments were well tolerated.


Witnessing history: the Infectious Diseases Society of Taiwan. The first twenty years.


A study of persistent viral infections using nude mice and a temperature-sensitive mutant of vesicular stomatitis virus.


title: Rapid Progression to Acute Respiratory Distress Syndrome: Review of Current Understanding of Critical Illness from COVID-19 Infection.

text: The coronavirus disease 2019 (COVID-19) outbreak that started in Wuhan, Hubei province, China in December 2019 has now extended across the globe with >100,000 cases and 3,000 deaths reported in 93 countries as of 7 March 2020. We report a case of COVID-19 infection in a 64-year-old man who developed rapidly worsening respiratory failure and acute respiratory distress syndrome (ARDS) that required intubation. As the clinical spectrum of COVID-19 ranges widely from mild illness to ARDS with a high risk of mortality, there is a need for more research to identify early markers of disease severity. Current evidence suggests that patients with advanced age, pre-existing comorbidities or dyspnoea should be closely monitored, especially at 1-2 weeks after symptom onset. It remains to be seen if laboratory findings such as lymphopenia or elevated lactate dehydrogenase may serve as early surrogates for critical illness or markers of disease recovery. Management of ARDS in COVID-19 remains supportive while we await results of drug trials. More studies are needed to understand the incidence and outcomes of ARDS and critical illness from COVID-19, which will be important for critical care management and resource planning.


title: Antigenic and biological relationships between human coronavirus OC43 and neonatal calf diarrhoea coronavirus.

text: Monospecific antisera were prepared in mice to human coronavirus OC43 and neonatal calf diarrhoea coronavirus (NCDCV) which had been previously adapted to growth in suckling mouse brain. Brain suspension from infected suckling mice was used as immunogen. The antigenic relationship between OC43 and NCDCV was studied by the indirect immunoperoxidase antibody technique, by the haemagglutination-inhibition (HI) test and a new infectious centre-reduction neutralization test. In mouse immune sera, a two-way cross-reaction between OC43 and NCDCV was detected. However, the antigenic relationship appeared to be closer for internal (as shown by immunoperoxidase staining) as compared to surface antigens (as shown by HI and neutralization). In primary infections of natural hosts there was a high degree of cross-reactivity between the two coronavirus strains for both surface and internal antigens, and homologous and heterologous titres were consistently within an eightfold dilution difference by all tests. Most human adults and calves had antibody to both OC43 and NCDCV and geometric mean titres of homologous antibody were higher than titres of heterologous antibody. Although OC43 and NCDCV share antigenic determinants, they possessed several different biological properties, including plaque morphology by the infectious centre assay, agglutination of 1-day-old chick erythrocytes and resistance of haemagglutinin to physical and chemical treatments.


title: Putative caveolin-binding sites in SARS-CoV proteins.

text: AIM To obtain the information of protein-protein interaction between the SARS-CoV proteins and caveolin-1, identify the possible caveolin-binding sites in SARS-CoV proteins. METHODS On the basis of three related caveolin-binding motifs, amino acid motif search was employed to predict the possible caveolin-1 related interaction domains in the SARS-CoV proteins. The molecular modeling and docking simulation methods were used to confirm the interaction between caveolin-1 and SARS-CoV proteins. RESULTS Thirty six caveolin-binding motifs in the SARS-CoV proteins have been mapped out using bioinformatics analysis tools. Molecular modeling and simulation have confirmed 8 caveolin-binding sites. These caveolin-binding sites located in replicase 1AB, spike protein, orf3 protein, and M protein, respectively. CONCLUSION Caveolin-1 may serve as a possible receptor of the SARS-CoV proteins, which may be associated with the SARS-CoV infection, replication, assembly, and budding.


title: Membrane fusion.

text: The fusion of biological membranes results in two bilayer-based membranes merging into a single membrane. In this process the lipids have to undergo considerable rearrangement. The nature of the intermediates that are formed during this rearrangement has been investigated. Certain fusion proteins facilitate this process. In many cases short segments of these fusion proteins have a particularly important role in accelerating the fusion process. Studies of the interaction of model peptides with membranes have allowed for increased understanding at the molecular level of the mechanism of the promotion of membrane fusion by fusion proteins. There is an increased appreciation of the roles of several independent segments of fusion proteins in promoting the fusion process. Many of the studies of the fusion of biological membranes have been done with the fusion of enveloped viruses with other membranes. One reason for this is that the number of proteins involved in viral fusion is relatively simple, often requiring only a single protein. For many enveloped viruses, the structure of their fusion proteins has certain common elements, suggesting that they all promote fusion by an analogous mechanism. Some aspects of this mechanism also appears to be common to intracellular fusion, although several proteins are involved in that process which is more complex and regulated than is viral fusion.


title: Opportunistic fungi and fungal infections: the challenge of a single, general antifungal vaccine.

text: A vaccine made up by an algal beta-glucan (laminarin), conjugated with a protein component, protects against infections by different fungi and induces antibodies capable of inhibiting fungal growth. Although taking a premium on a common molecular theme, this remains a sort of 'cross-kingdom' vaccine because the immunizing antigen and the vaccination target belong to organisms from two different kingdoms and this is certainly the first case in the field of human vaccines. Thus, it is possible to convey in a single immunological tool the potential to protect against multiple infections, in theory all those caused by beta-glucan-expressing fungi. The generation of antibodies with the potential to directly inhibit the growth of, or kill the fungal cells also opens an exciting perspective for both active and passive vaccination in immunocompromised subjects.


title: An overview of immunological and genetic methods for detecting swine coronaviruses, transmissible gastroenteritis virus, and porcine respiratory coronavirus in tissues.

text: Transmissible gastroenteritis (TGE) is an enteric disease of swine caused by a coronavirus, designated as transmissible gastroenteritis virus (TGEV). Commonly used methods for TGEV detection include viral isolation and detection of the viral antigen by indirect immunofluorescence (IFA), immunoperoxidase, and immunogold silver staining. Each of these techniques has some advantages and disadvantages. In general IFA and immunohistochemistry are preferred over viral isolation as TGEV isolation is not very reliable because not all field isolates replicate in cell cultures. The diagnosis of TGEV has become more complicated since the emergence of porcine respiratory coronavirus (PRCV). PRCV is believed to be a TGEV mutant, and can not be easily differentiated from TGEV by immunological tests. Nucleic acid probes and polymerase chain reaction (PCR) have successfully been used to detect and differentiate these viruses. These techniques can detect viral nucleic acids in the specimen but do not provide information on the cell types infected by these viruses. Recently we have developed isotopic and nonisotopic in situ hybridization techniques (ISH) for the detection of these viral nucleic acids in formalin-fixed paraffin-embedded tissues. Furthermore, this procedure can differentiate between TGEV- and PRCV-infected cells. By ISH, TGEV is detected in the mature absorptive enterocytes of tissues infected by TGEV and the crypt epithelial cells are also infected but to a lesser extent. For PRCV, the main infected cells are epithelial cells of the bronchioles, type II pneumocytes, and alveolar and septal macrophages. ISH is an excellent tool for studying molecular pathogenesis of these two viruses especially when used in combination with immunohistochemistry.


Monogamy is...


title: [Glucocorticoid in the treatment of severe acute respiratory syndrome patients: a preliminary report].

text: OBJECTIVE To discuss the effect of glucocorticoid (methylprednisolone) on severe acute respiratory syndrome (SARS). METHODS Thirty SARS patients were treated at our hospital for over 3 weeks since March to May 2003. The course and dosage of glucocorticoid, counts of CD(4)(+), CD(8)(+) and CD(3)(+), electrolytes, blood routine, and sera albumin before and after the treatment were analysed. RESULTS Before treatment by methylprednisolone, the counts of CD(4)(+), CD(8)(+) and CD(3)(+) of 27 SARS patients were (401 +/- 203), (340 +/- 187), (756 +/- 383) cells/ microl. Twenty-nine of the 30 SARS patients were treated by methylprednisolone. The dosage for 24 patients was 80 - 160 mg/d with the largest being 1,000 mg/d before admission to the hospital. The count of WBC was increased after treatment (P < 0.01). No obvious effect was observed on the potassium, sodium and chlorine of blood (P > 0.05). Glucocorticoid increased the level of blood glucose (P = 0.01), decreased the level of sera albumin (P < 0.01), and its large dosage decreased the counts of CD(4)(+), CD(8)(+) and CD(3)(+). Three severe patients had secondary infection after administration of a large dose of glucocorticoid. CONCLUSIONS In the early stage of the disease, the counts of CD(4)(+), CD(8)(+) and CD(3)(+) of SARS patients may reduce markedly indicating the immunity is suppressed. A large dose of glucocorticoid may aggravate the suppression and make the body in an active metabolic state (the increase of blood glucose and the decrease of sera albumin). Thus the disease is aggravated and patients are likely to suffer from severe secondary infection. Indications for use of glucocorticoid must strictly controlled and its large dosage is improper.


title: [Oligonucleotide microarray preparation using enhanced poly-L-lysine glass slides].

text: OBJECTIVE To modify conventional poly-L-lysine coating for oligonucleotide microarray preparation so as to enhance the sensitivity of the microarray. METHOD The proposed chemical approaches included silanizing the slides with 3-glycid-oxypropyltrimethoxysilane (GOPS) after cleaning, followed by slide coating with polymers (poly-L-lysine) that was covalently bound to the modified glass. Subsequent attachment of the oligonucleotide to the modified slide surface was achieved after 1,4-phenylene diisothiocyanate (PDITC) activation of the surface. Various experiments were carried out, such as the immobilization efficiency and hybridization assays to test the modified slides, which were then used tentatively in the preparation of microarrays for SARS coronavirus detection. RESULTS The improved surface had high immobilization efficiency, good uniformity and satisfactory hybridization efficiency, better than those slides with conventional poly-L-lysine coating. In addition, such modified slides rendered the microarrays more resistant to consecutive probing/stripping cycles. CONCLUSION The modified slide surface is satisfactory to immobilize unmodified oligonucleotide by covalent binding, which enhances not only the sensitivity of the prepared oligonucleotide microarray but also the binding of the oligonucleotide to the slide surface.


title: The Role of Conventional Ultrasonography in the Evaluation of Antrum Wall Thickness in Obese Patients.

text: BACKGROUND The objective of this study is to evaluate the utility of conventional ultrasonography (USG) in the evaluation of the stomach antrum and distal corpus lesions. METHODS A prospective evaluation was made of 69 patients who underwent sleeve gastrectomy. Preoperative USG was applied to the patients and measurements were taken and recorded of the stomach antrum full layer wall thickness (USGFT) and of mucosal thickness (USGMT). Postoperatively, same parameters were again measured histopathologically and the pathological full thickness (PFT) and pathological mucosal thickness (PMT) values were compared. RESULTS When evaluation was made in respect of USG and pathological measurements, the USGFT was 8.51 ± 3.07 (range 4.5-15.8) and USGMT was 5.80 ± 2.15 (range 2.36-10.5). The PFT was determined as 8.13 ± 2.24 (range 4-14) and PMT as 5.53 ± 1.86 (range 2-10.5). In the histopathological examination, gastritis was seen in 53 (76.8 %) patients and Helicobacter pylori (HP) positivity was determined in 32 (46.4 %) patients. When the patients were grouped as obese (BMI ≤ 49.9 kg/m2) (group 1, n = 50) and super obese (BMI ≥ 50 kg/m2) (group 2, n = 19), no difference was determined between the groups ultrasonographically or histopathologically (p > 0.05). The antrum wall thickness was seen to be significantly greater in the patients with gastritis and HP positivity compared to the patients who were negative. In ROC analysis, cutoff values were calculated for USGFT (5.86 mm) and USGMT (4.49 mm). In gastritis diagnosis, the USGFT cutoff value was found to have 796 % sensitivity and 68.7 % specificity. CONCLUSION USG was seen to be an extremely effective method in visualising the antrum wall and gastritis diagnosis can be made comfortably from the wall thickness measurement.


title: Glial grafting for demyelinating disease.

text: Remyelination of demyelinated central nervous system (CNS) axons is considered as a potential treatment for multiple sclerosis, and it has been achieved in experimental models of demyelination by transplantation of pro-myelinating cells. However, the experiments undertaken have not addressed the need for tissue-type matching in order to achieve graft-mediated remyelination since they were performed in conditions in which the chance for graft rejection was minimized. This article focuses on the factors determining survival of allogeneic oligodendrocyte lineage cells and their contribution to the remyelination of demyelinating CNS lesions. The immune status of the CNS as well as the suitability of different models of demyelination for graft rejection studies are discussed, and ways of enhancing allogeneic oligodendrocyte-mediated remyelination are presented. Finally, the effects of glial graft rejection on host remyelination are described, highlighting the potential benefits of the acute CNS inflammatory response for myelin repair.


title: The widening scope of coronaviruses.

text: PURPOSE OF REVIEW In the past 2 years, at least three distinct human coronaviruses have been discovered, including the etiological agent associated with severe acquired respiratory syndrome (SARS). These recently discovered viruses, with the exception of the SARS associated coronavirus (SARS-CoV), are likely to be common respiratory viruses and may be responsible for a substantial proportion of respiratory tract disease. RECENT FINDINGS The SARS-CoV first appeared in 2002 and spread rapidly around the globe. Although the worldwide spread of SARS-CoV may have been halted, the emergence of this new virus demonstrates the potential threat represented by species-to-species transmission of coronaviruses. NL63, initially isolated from a young child with lower respiratory tract disease, represents a group of newly described group I coronaviruses that have been identified worldwide, which are associated with both upper and lower respiratory tract disease, particularly in young children. The distribution of HKU1, a newly identified group II coronavirus, is not yet established. NL63 and HKU1 are related to the common human coronaviruses 229E and OC43, respectively. SUMMARY The discovery of at least three new human coronaviruses represents significant advances in the investigation of human respiratory tract disease. Further studies are required to fully define the impact of these new pathogens.


title: [SARS surveillance--did it meet the demands made on the surveillance of new infectious diseases?].

text: INTRODUCTION The world-wide outbreak of severe acute respiratory syndrome (SARS) in the spring of 2003 highlighted the importance of reliable surveillance for infectious diseases. The aim of this investigation is to identify possibilities for the improvement of the German surveillance system based on the experiences made during the SARS epidemic 2003. METHODS The RKI has established two linked databases for the surveillance of SARS, to store case-based data on SARS and to keep a record of inquiries by the public health system. Based on these data we evaluated the information flow for the surveillance of SARS mainly qualitatively. RESULTS AND DISCUSSION With occurrence of the first SARS case in Germany a nationwide surveillance system for SARS was established. Since Germany was exclusively confronted with imported SARS cases, the surveillance system could fulfil the requirements. It is unclear, however, whether the current procedures are sufficient to manage larger epidemics of emerging diseases. For this task the public health system needs a uniform electronic information management system which allows rapid adaptation to the epidemic situation and is able to deal with a large amount of data.


title: Mitochondrial aconitase binds to the 3' untranslated region of the mouse hepatitis virus genome.

text: Mouse hepatitis virus (MHV), a member of the Coronaviridae, contains a polyadenylated positive-sense single-stranded genomic RNA which is 31 kb long. MHV replication and transcription take place via the synthesis of negative-strand RNA intermediates from a positive-strand genomic template. A cis-acting element previously identified in the 3' untranslated region binds to trans-acting host factors from mouse fibroblasts and forms at least three RNA-protein complexes. The largest RNA-protein complex formed by the cis-acting element and the lysate from uninfected mouse fibroblasts has a molecular weight of about 200 kDa. The complex observed in gel shift assays has been resolved by second-dimension sodium dodecyl sulfate-polyacrylamide gel electrophoresis into four proteins of approximately 90, 70, 58, and 40 kDa after RNase treatment. Specific RNA affinity chromatography also has revealed the presence of a 90-kDa protein associated with RNA containing the cis-acting element bound to magnetic beads. The 90-kDa protein has been purified from uninfected mouse fibroblast crude lysates. Protein microsequencing identified the 90-kDa protein as mitochondrial aconitase. Antibody raised against purified mitochondrial aconitase recognizes the RNA-protein complex and the 90-kDa protein, which can be released from the complex by RNase digestion. Furthermore, UV cross-linking studies indicate that highly purified mitochondrial aconitase binds specifically to the MHV 3' protein-binding element. Increasing the intracellular level of mitochondrial aconitase by iron supplementation resulted in increased RNA-binding activity in cell extracts and increased virus production as well as viral protein synthesis at early hours of infection. These results are particularly interesting in terms of identification of an RNA target for mitochondrial aconitase, which has a cytoplasmic homolog, cytoplasmic aconitase, also known as iron regulatory protein 1, a well-recognized RNA-binding protein. The binding properties of mitochondrial aconitase and the functional relevance of RNA binding appear to parallel those of cytoplasmic aconitase.


Coronavirus induced demyelinating encephalomyelitis in rats: immunopathological aspects of viral persistency.


title: An electron microscopic study of viruses associated with canine gastroenteritis.

text: An electron microscopic study was carried out on specimens of feces and intestinal contents from cases of canine gastroenteritis submitted to the Diagnostic Laboratory, New York State College of Veterinary Medicine, during 1979-1981. The majority of samples came from New York State and the Northeast with no marked shift in distribution over the three year period. Canine parvovirus was the major virus identified. In August and September 1980 there was an epidemic of canine gastroenteritis, with 247 samples received during this two month period alone, of which 48 percent were positive for canine parvovirus. Almost half of the total number of specimens examined were from dogs less than 6 months of age and well over 50 percent of these were parvovirus positive. In addition to canine parvovirus, three cases of coronavirus, two cases of rota-like virus and one case of astro-like virus were detected. Three dual infections with canine parvovirus and rota or astro-like virus were also confirmed. An unidentified virus-like particle with cubic symmetry was found in two specimens. The adoption of immunoelectron microscopy for the detection of canine parvovirus in March 1980 facilitated identification of this virus and greatly increased the sensitivity of the technique.


title: [The effect of platelet-rich plasma on graft healing in reconstruction of the anterior cruciate ligament of the knee joint: prospective study].

text: PURPOSE OF THE STUDY Growth factors produced by platelets enhance tissue healing. The aim of this study was to confirm or disprove the hypothesis that, in anterior cruciate ligament (ACL) reconstruction, the application of platelet-rich plasma (PRP) into the tibial and femoral tunnels and in the graft enhances graft maturation and graft-bone interface healing and thus improves knee function at 3 and 12 months post-operatively in comparison with the control group. MATERIAL AND METHODS A total of 40 patient had the surgery; 20 underwent single-bundle hamstring reconstruction with PRP application (PRP group) and 20 had the same surgery without PRP addition (control group). A 5 ml amount of PRP was obtained from the patient's peripheral blood. A graft inserted in the bone tunnels was fixed with interference screws and, after intra-articular fluid aspiration, 1 ml of PRP was injected into each tunnel and 3 ml were evenly applied to the intra-articular portion of the graft. The patients were examined by MRI at 3 and 12 months after surgery. The subsidence of swelling in the tunnelsurrounding tissues was taken as a sign of graft-bone interface healing, and increased signal intensity of the graft was considered as a result of its ligamentisation. The knee functional status was evaluated at 3 and 12 post-operative months, using the scoring systems (Cincinnati score, IKDC score). RESULTS Bone swelling was found at 3 post-operative months in 18 of 20 patients in both the PRP and the control group. Graft signal intensity was increased in most patients (19 of the PRP group; 18 control patients; p = 0.949). The Cincinnati score at 3 months had an average value of 72.7 (34-100; SO, 18.7) in the PRP group and 73.4 (42-99; SO, 16.3) in the control group (p = 0.793). The functional score after 12 months improved to 97.5 (75-100; SO, 12.8) in the PRP group and to 95.1 (66-100; SO, 13.1) in the control group; there was no significant difference between the groups (p = 0.885) at either 3 or 12 months. The IKDC score showed similar results. At 12 months bone swelling was recorded in seven out of 20 patients in the PRP group and in nine patients in the control group (p = 0.751). Graft signal intensity was increased in three and four patients of the PRP and control groups, respectively (p = 0.681). There was a statistical difference between the findings at 3 and 12 months within each group, but no difference between the groups. DISCUSSION In ACL reconstruction, the process of tendon graft-to-bone healing has several stages involving inflammation, cell proliferation and graft ligamentisation. At each stage, an important role is played by growth factors produced by thrombocytes, and therefore their potential use in the treatment of injuries to ligaments and tendons has recently come into focus. A number of experimental studies dealing with the effect of platelet-rich plasma on soft tissue healing has been published. Therefore, the method of using PRP to enhance graft healing in ACL reconstruction in humans is still being studied. CONSLUSIONS The use of PRP in ACL reconstruction does not accelerate graft remodelling and bone ingrowth into the tendon. The function scoring results showed a statistically significant improvement in knee function between 3 and 12 months of followup within each group studied but revealed no difference between the groups. The hypothesis postulated above was disproved. Key words: anterior cruciate ligament, platelet-rich plasma, anatomic single-bundle reconstruction.


title: Transmissible enteritis of turkeys: experimental inoculation studies with tissue-culture-adapted turkey and bovine coronaviruses.

text: Four Quebec isolates of turkey enteric coronaviruses (TCVs) and three isolates of bovine enteric coronaviruses (BCVs) were serially propagated in HRT-18 and compared for their pathogenicity in turkey embryos and turkey poults. By immunoelectron microscopy, hemagglutination-inhibition, and Western immunoblotting assays, tissue-culture-adapted Quebec TCV isolates were found to be closely related to the reference Minnesota strain of TCV and the Mebus strain of BCV. Genomic relationships between TCV isolates and the reference BCV strain were confirmed by hybridization assays with BCV-specific radiolabeled recombinant plasmids containing sequences of the N and M genes. Only TCV isolates could be propagated by inoculation in the amniotic cavity of chicken and turkey embryonating eggs, and induced clinical disease in turkey poults. Nevertheless, coronavirus particles or antigens were detected by electron microscopy or indirect enzyme-linked immunosorbent assay in the clarified intestinal contents of BCV-infected poults up to day 14 PI, and genomic viral RNA was detected by slot-blot hybridization using BCV cDNA probes.


title: Novel coronavirus: the challenge of communicating about a virus which one knows little about.

text: Following the severe acute respiratory syndrome (SARS) event in 2002/2003, the Worl Health Organization (WHO) developed outbreak communications guidelines. With the emergence in September 2012 of a novel coronavirus, WHO's public communications response was initiated and planned in light of these guidelines and 5 principles of trust, transparency, announcing early, listening and planning. This review describes WHO's communication response to the novel coronavirus event'and its efforts to provide early, accurate information via various media to keep the public appraised of the situation, and its commitment to continued communication on an ongoing basis.


title: Enteric infections with coronaviruses and toroviruses.

text: Many enteric viruses are difficult or impossible to propagate in tissue culture. Coronaviruses and toroviruses are large, enveloped, plus-strand RNA viruses in the order Nidovirales that cause enteric disease in young pigs, cows, dogs, mice, cats and horses. Two different serogroups of mammalian coronaviruses cause frequent respiratory infections in humans, and coronaviruses and toroviruses have been implicated in human diarrhoeal disease by immunoelectron microscopy. However, there is as yet no consensus about the importance of these enveloped viruses in human diarrhoea, and little is known about their genetic variability. The large spike (S) glycoprotein is an important determinant of species specificity, tissue tropism and virulence of coronavirus infection. To infect enterocytes, both S glycoproteins and the viral envelope must resist degradation by proteases, low and high pH, and bile salts. One specific site on the S glycoprotein of bovine coronavirus must be cleaved by an intracellular protease or trypsin to activate viral infectivity and cell fusion. S glycoprotein binds to specific receptors on the apical membranes of enterocytes, and can undergo a temperature-dependent, receptor-mediated conformational change that leads to fusion of the viral envelope with host membranes to initiate infection. Analysing spike-receptor interactions may lead to new ways to propagate these enteric viruses as well as new strategies for development of novel antiviral drugs.


title: Sensitivity and specificity of a nested polymerase chain reaction for detection of lentivirus infection in lions (Panthera leo).

text: Feline immunodeficiency virus (FIV) is a lentivirus in the Retroviridae family that causes lifelong infection in domestic cats. The lentivirus of African lions (Panthera leo), referred to as FIVple, is endemic in certain lion populations in eastern and southern Africa. Lentivirus infection leads to immunologic dysfunction and immunosuppressive disease in domestic cats; however, little is known about the pathogenic effects of infection in lions, nor about the epidemiologic impact on free-ranging and captive populations. Whole blood and serum samples were collected opportunistically from free-ranging lions in Kruger National Park, Republic of South Africa (RSA). Whole blood and serum samples were also collected from captive wild lions in the RSA. A nested polymerase chain reaction (PCR) assay for detection of FIV was performed on all whole blood samples. In addition, serum samples were tested for cross-reactive antibodies to domestic feline lentivirus antigens and puma lentivirus synthetic envelope peptide antigen. The PCR assay successfully amplified the lion lentivirus from African lions. The relative sensitivity and relative specificity were 79% and 100%, respectively, and the positive and negative predictive values were 100% and 67%, respectively. This research represents the first study to compare genetic material with antibody-based methods of lentivirus detection on lions in RSA. Using PCR as an additional diagnostic test for FIV in lions will increase screening sensitivity and will allow viral characterization among circulating isolates and monitoring of changes in the viral epidemiology within geographic regions and populations over time.


title: CD13/APN regulates endothelial invasion and filopodia formation.

text: CD13/aminopeptidase N is a transmembrane peptidase that is induced in the vasculature of solid tumors and is a potent angiogenic regulator. Here, we demonstrate that CD13 controls endothelial cell invasion in response to the serum peptide bradykinin by facilitating signal transduction at the level of the plasma membrane. Inhibition of CD13 abrogates bradykinin B(2) receptor internalization, leading to the attenuation of downstream events such as bradykinin-induced activation of Cdc42 and filopodia formation, and thus affects endothelial cell motility. Investigation into mechanisms underlying this block led us to focus on B(2)R internalization via membrane-dependent mechanisms. Membrane disruption by depletion of cholesterol or trypsinization halts B(2)R internalization, invasion, and filopodia formation, which can be recovered with addition of cholesterol. However, this functional recovery is severely impaired in the presence of CD13 antagonists, and the distribution of membrane proteins is disordered in treated cells, suggesting a role for CD13 in plasma membrane protein organization. Finally, exogenous expression of wild-type but not mutant CD13 further alters protein distribution, suggesting peptidase activity is required for CD13's regulatory activity. Therefore, CD13 functions as a novel modulator of signal transduction and cell motility via its influence on specific plasma membrane organization, thus regulating angiogenesis.


title: Protecting human and ecological health under viral threats in Asia.

text: Severe acute respiratory syndrome (SARS) outbroke in 2003, and the avian influenza A (H5N1) also outbroke in 2003 and continued to 2004. These pandemic viral diseases originated in South East Asia. Many human and animal lives were lost. Economic damages due to the pandemics were also very large. The question arises of why did the pandemics originate from South East Asian areas. Human influenza A consists of many sub-types of coronaviruses including the SARS virus and the avian influenza (H5N1) that are all variants of RNA of avian coronavirus. Variants are formed during infection of a coronavirus through not only birds but also mammals, including human beings. There are hot spots where viral infection rates are accelerated among birds, mammals and human beings. Suspicious areas are in South East Asia, where living conditions of birds, mammals and human beings are so close that there are always risks of viral infection. When we see the living conditions of farmers in southern China, northern Vietnam, Laos and northern Myanmar, they commonly raise ducks/chickens with pigs sharing ponds into which they discharge household wastewater, including human excreta, and pig excreta that are significant carriers of viruses. Bird faeces are also key carriers of the viruses. In the ponds, they raise ducks and conduct fish culture. Other important players are migrating birds from North Asia, which are principal vectors of avian influenza viruses. There is an urgent necessity of improving human and ecological health in South East Asia to control viral infection among birds, mammals and human beings. We can hinder the vicious cycle of virus infection through water contamination in ponds by providing good human, pig and chicken sanitation. It is easy to provide good sanitation practices for human, pigs and chickens, introducing collection and treatment of excreta. Our modern water technology can find good solutions for the problem.


Structure and expression of the bovine coronavirus hemagglutinin protein.


title: Apical plasma membrane proteins are not obligatorily stored in secretory granules in exocrine cells.

text: Exocrine cells are epithelial cells in which secretory granules undergo fusion with the apical plasma membrane upon secretagogue stimulation. Several apical plasma membrane proteins have been found in secretory granules in cells from pancreas and salivary glands raising the possibility that incorporation into secretory granules followed by exocytosis of the granules accounts for their insertion into the apical plasma membrane. To test this hypothesis, we have expressed the influenza hemagglutinin (HA) in pancreatic AR42J cells, which make zymogen-like granules upon incubation with dexamethasone. The influenza virus HA is known to be specifically targeted to the apical plasma membrane of epithelial cells that lack a regulated pathway and is also known to be excluded from secretory granules in virally-infected pituitary AtT20 cells. Localization of the protein by immunofluorescence microscopy revealed that it accumulated at the plasma membrane of the transfected AR42J cells. HA was not observed in the amylase-rich secretory granules. By immunolabeling of ultrathin cryosections of the transfected cells, HA was also found exclusively on the cell surface, with label over secretory granules not exceeding that seen in control, untransfected cells. In addition, in cell fractionation experiments performed on radiolabeled AR42J cell transformants, HA was not detectable in the secretory granule fractions. These results indicate that HA is not efficiently stored in mature secretory granules and is likely to reach the cell surface via constitutive transport pathways.


title: Management of hospital-acquired severe acute respiratory syndrome with different disease spectrum.

text: BACKGROUND Outbreak of severe acute respiratory syndrome (SARS) in Taipei has been associated with Taiwanese back from Guangdong, China. We report 4 probable SARS cases with different severity and propose optimal treatment. METHODS Four probable SARS cases were enrolled. Two cases were due to outbreak of SARS in our hospital and two cases were transferred from other hospitals. All patients received standard treatment: ribavirin 1000 mg orally daily for 10 days, Levofloxacin 500 mg orally daily for 7 days, and intravenous immunoglobulin (IVIG) 1 g/kg/day for 2 day after the onset of symptoms. If severe hypoxia (PaO2/FiO2 < 200) occurred, protective strategy of mechanical ventilation and methylprednisolone 2 mg/kg/day were given. The clinical pictures and treatment outcome were discussed. RESULTS Fever, dyspnea, diarrhea, malaise, dizziness and dry cough were initially more common symptoms. Initially chest patterns included focal consolidation, interstitial infiltration or normal. Common laboratory findings were lymphopenia, and elevated serum levels of lactate dehydrogenase and C-reactive protein. No mortality was found. CONCLUSIONS Highly alert and stringent infection control of SARS cases are required. Otherwise, SARS easily induces hospital-acquired first then community-acquired infection. Initial presentation of radiographic patterns includes normal, interstitial or airspace shadowing. Fever and lymphopenia are occasionally followed by rapidly progressive respiratory compromise. The standard treatment might be beneficial for decreasing the mortality rate.


title: Stem-loop IV in the 5' untranslated region is a cis-acting element in bovine coronavirus defective interfering RNA replication.

text: The 210-nucleotide (nt) 5' untranslated region (UTR) in the positive-strand bovine coronavirus (BCoV) genome is predicted to contain four higher-order structures identified as stem-loops I to IV, which may function as cis-acting elements in genomic RNA replication. Here, we describe evidence that stem-loop IV, a bulged stem-loop mapping at nt 186 through 215, (i) is phylogenetically conserved among group 2 coronaviruses and may have a homolog in groups 1 and 3, (ii) exists as a higher-order structure on the basis of enzyme probing, (iii) is required as a higher-order element for replication of a BCoV defective interfering (DI) RNA in the positive but not the negative strand, and (iv) as a higher-order structure in wild-type (wt) and mutant molecules that replicate, specifically binds six cellular proteins in the molecular mass range of 25 to 58 kDa as determined by electrophoretic mobility shift and UV cross-linking assays; binding to viral proteins was not detected. Interestingly, the predicted stem-loop IV homolog in the severe acute respiratory syndrome (SARS) coronavirus appears to be group 1-like in that it is in part duplicated with a group 1-like conserved loop sequence and is not group 2-like, as would be expected by the SARS coronavirus group 2-like 3' UTR structure. These results together indicate that stem-loop IV in the BCoV 5' UTR is a cis-acting element for DI RNA replication and that it might function through interactions with cellular proteins. It is postulated that stem-loop IV functions similarly in the virus genome.


title: [Comparative study of clinical characteristics and prognosis of clinically diagnosed SARS patients with positive and negative serum SARS coronavirus-specific antibodies test].

text: OBJECTIVE To investigate clinical characteristics and long-term effects of SARS. METHODS Clinical characteristics of 197 SARS patients in Xiao Tang-shan hospital were analyzed retrospectively, and prognosis of them were analyzed prospectively. RESULTS Among the 197 patients, 153 patients (77.7%) have positive results of serum SARS coronavirus-specific antibodies test, and 44 patients (22.3%) have negative results. The average age of SARS and non-SARS patients were (40 +/- 12) and (31 +/- 12) years, male/female ratio were 1.0:1.6 and 2.1:1.0, average body temperature were (38.5 degrees C +/- 0.3 degrees C) and (38.1 degrees C +/- 0.4 degrees C), median of fever length were 7.0 d (0.4 approximately 50 d) and 2.3 d (0.3 approximately 37 d) respectively. The occurrence of dyspnea, malaise and gastrointestinal symptom were more often in SARS patient than in non-SARS patients. Some patients have residual symptoms (such as cough, fatigue, dyspnea, abnormality of liver function, hyperglycemia and hyperglyceridemia), and only few patients have lung fibrosis. CONCLUSION Some patients with other respiratory diseases were misdiagnosed as SARS. There were several obvious differences of clinical characteristics between SARS patient and non-SARS patients. Prognosis of most patients were preciously well, and few still had abnormality of lung function. Residual symptoms of SARS and side effects of drugs used to treat SARS should be discriminated.


title: Iodine Concentration and Optimization in Computed Tomography Angiography: Current Issues.

text: Computed tomography (CT) technology has seen a dramatic evolution in the recent past that has deeply changed the face of this diagnostic modality. Since the early days of helical single-slice and then multislice CT, CT angiography (CTA) has been one of the most technically demanding applications, both in terms of scanning technique and contrast medium (CM) injection protocol, due to the need to acquire a large amount of high-resolution data over a limited period corresponding to the peak contrast enhancement of the arterial system. Iodine concentration is one of the main determinants of arterial enhancement in CTA, and current low-osmolar and iso-osmolar nonionic CM for intravascular administration still come in a handful of molecules, but a relatively wide range of different iodine concentrations. This gives the opportunity to optimize CTA protocols as a function of several factors such as patient characteristics, CT technology, and CM features in an attempt to maximize the diagnostic yield of CTA examinations while considering patient safety and avoiding unnecessary extra costs. Our aim is to provide an up-to-date overview of the existing evidence on how changing iodine concentration can have an impact on CTA performance, especially with the use of state-of-the-art CT and power injector technology, in the perspective of improving patient care while minimizing overall exposure to iodinated CM and ionizing radiation.


Clinical data marshalled to treat HIV.