title: [Clinical features of asymptomatic or subclinical COVID-19 in children].

text: OBJECTIVE To study the clinical features of asymptomatic or subclinical coronavirus disease 2019 (COVID-19) in children. METHODS A retrospective analysis was performed for the clinical data of 53 children who were confirmed with asymptomatic or subclinical COVID-19, including epidemiological history, clinical typing, co-infection, time to clearance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleic acid in nasopharyngeal swabs, laboratory examination results, length of hospital stay, and treatment outcome. RESULTS The children with asymptomatic or subclinical COVID-19 accounted for 30.5% (53/174) in children with COVID-19 hospitalized in the COVID-19 ward of Wuhan Children's Hospital. All cases occurred with familial aggregation. Among the 53 children, 35 (66%) had asymptomatic infection and 18 (34%) had subclinical infection. Mycoplasma infection was found in 17 children (32%). For the 53 children, the mean time to clearance of SARS-CoV-2 nucleic acid in nasopharyngeal swabs was 9±4 days. Most laboratory markers were maintained within the normal range. The mean hospital stay was 11±4 days. Lung CT of 18 children with subclinical COVID-19 showed ground-glass opacities, linear opacities, and patchy opacities, with relatively limited lesions. CONCLUSIONS There is a high proportion of children with asymptomatic or subclinical COVID-19 among the children with COVID-19 hospitalized in the COVID-19 ward. The transmission risk of asymptomatic or subclinical COVID-19 should be taken seriously.


title: Hepatitis C virus glycoproteins mediate low pH-dependent membrane fusion with liposomes.

text: It has been suggested that the hepatitis C virus (HCV) infects host cells through a pH-dependent internalization mechanism, but the steps leading from virus attachment to the fusion of viral and cellular membranes remain uncharacterized. Here we studied the mechanism underlying the HCV fusion process in vitro using liposomes and our recently described HCV pseudoparticles (pp) bearing functional E1E2 envelope glycoproteins. The fusion of HCVpp with liposomes was monitored with fluorescent probes incorporated into either the HCVpp or the liposomes. To validate these assays, pseudoparticles bearing either the hemagglutinin of the influenza virus or the amphotropic glycoprotein of murine leukemia virus were used as models for pH-dependent and pH-independent entry, respectively. The use of assays based either on fusion-induced dequenching of fluorescent probes or on reporter systems, which produce fluorescence when the virus and liposome contents are mixed, allowed us to demonstrate that HCVpp mediated a complete fusion process, leading to the merging of both membrane leaflets and to the mixing of the internal contents of pseudoparticle and liposome. This HCVpp-mediated fusion was dependent on low pH, with a threshold of 6.3 and an optimum at about 5.5. Fusion was temperature-dependent and did not require any protein or receptor at the surface of the target liposomes. Most interestingly, fusion was facilitated by the presence of cholesterol in the target membrane. These findings clearly indicate that HCV infection is mediated by a pH-dependent membrane fusion process. This paves the way for future studies of the mechanisms underlying HCV membrane fusion.


title: Video-assisted thoracoscopic surgery for the central bronchogenic carcinoma with invasion of the main bronchus and pulmonary artery: A case report.

text: Surgical treatment for central bronchogenic carcinoma with invasion of the main bronchus and pulmonary artery often include bronchial anastomosis and pulmonary angioplasty. As a technique, video-assisted thoracoscopic surgery (VATS) lobectomy has proven to reduce the rate of occurrence of complications, and thereby obtain improved survival rates. In the present case study, its use in treating central bronchogenic carcinoma with invasion of the main bronchus and pulmonary artery is reported. A case study of a 68-year-old man with a history of smoking 15-20 cigarettes per day for 40 years is described, who presented with a cough and hemoptysis. A bronchial sleeve left upper lobectomy and pulmonary angioplasty were performed with complete VATS and routine lymph node dissection. The preoperative diagnosis of squamous cell carcinoma (SCC) of the lung (cT2, N2, M0, stage IIIA) was confirmed as SCC through the postoperative pathological examination, leading to the tumor staging: pT2, N1, M0, stage IIB. These results were obtained without sacrificing the oncological principles of thoracic surgery. It has been demonstrated that VATS may be applied for treating central bronchogenic carcinoma with invasion of the main bronchus and pulmonary artery. However, end-to-end bronchial anastomosis and continuous suture of the pulmonary artery were difficult to perform, and use of the VATS procedure is on a learning curve, which will be informative for surgeons and their assistants.


title: Basic Shoulder Arthroscopy: Lateral Decubitus Patient Positioning.

text: Shoulder arthroscopy offers a minimally invasive surgical approach to treat a variety of shoulder pathologies. The patient can be positioned in either the lateral decubitus or the beach chair position. This note and accompanying video describe the operating room setup for shoulder arthroscopy in the lateral decubitus position, including positioning of the arms, head, and sterile preparation and draping. Appropriate lateral decubitus positioning for shoulder arthroscopy with careful attention to detail will promote ease of surgical intervention and minimize complications.


title: A resource-based perspective on retention strategies for nurse epidemiologists.

text: AIM This paper is a report of a study to evaluate the intent of epidemiologists to remain in their chosen career and identify the variables that contributed to or predicted their intent to stay. BACKGROUND Recently, emerging new infections, such as severe acute respiratory syndrome or bird flu, have placed significant occupational and psychological stress on epidemiologists, especially in South-East Asia, resulting in a high intent to change their career. In the light of possible staff shortages, retention strategies for epidemiologists have gained importance. METHODS A self-administered questionnaire survey and stratified sampling were used to collect data from 351 epidemiologists including nurse epidemiologists in Taiwan in 2005; response rate was 70.6%. Correlation analysis and hierarchical multiple regression analysis were used to examine relationships among occupational stress, psychological stress, human resources and intent to stay in their career. RESULTS Occupational stress, psychological stress and human resources had an impact on epidemiologists' intent to stay in their career. Results show that the relationship between occupational stress (operation and personal safety hazard) and intent to stay could be influenced by organizational capital, and the relationship between emotional distress and intent to stay could be influenced by a broad spectrum of human resources (organizational, social and human capital). CONCLUSION The severe acute respiratory syndrome epidemic raised worldwide attention and challenged epidemiologists' intent to stay. Results indicate that human resources play an important role in this issue. Managers should enhance human resources in organizations as much as possible to attenuate epidemiologists' stress, which may, in turn, strengthen their intent to stay.


title: Influence of thyroidectomy on postoperative serum calcium level regarding serum vitamin D status. A prospective study.

text: BACKGROUND Hypocalcemia is a well-recognized complication after total thyroidectomy. Hypovitaminosis D may have additional effect in the development of hypocalcemia. This study aimed to determine the effect of total thyroidectomy on postoperative serum calcium in patients with and without hypovitaminosis D. METHODS This prospective study was performed on patients who underwent total thyroidectomy from 2011 to 2014 in Imam Khomeini General Hospital of Mazandaran University of Medical Sciences. Serum calcium and vitamin D values were recorded before and after surgery. The patients were classified according to serum vitamin D concentrations as less 10 ng/ml (vitamin D deficiency) or higher (control group). The mean values of postoperative calcium level for each class of serum vitamin D were determined and compared. Hypocalcemia was defined as a postoperative calcium level <8 mg/dl. RESULTS 125 patients due to thyroid disease underwent total thyroidectomy. The incidence of symptomatic and asymptomatic hypocalcemia after surgery was 12% (n=15) and 3.2% (n=4) respectively. 82 (65.6%) patients had vitamin D deficiency and 43 (34.4%) patients had sufficient vitamin D level. There was not any significant difference in calcium level (8.67±0.58 mg/dl vs. 8.70±0.59 mg/dl) between two vitamin D studied groups after thyroid surgery (p>0.05). CONCLUSION The findings of this study indicated that vitamin D deficiency had no significant effect on post-thyroidectomy serum calcium level.


title: Distribution and trafficking of JHM coronavirus structural proteins and virions in primary neurons and the OBL-21 neuronal cell line.

text: The neurotropic murine coronavirus JHM is capable of inducing various forms of neurologic diseases, including demyelination. Neurons have been shown to act as a repository site at the early stages of the disease process (O. Sorensen and S. Dales, J. Virol. 56:434-438, 1985). JHM virus (JHMV) replication and trafficking of viral proteins and virions in cultured rat hippocampal neurons and a neuronal cell line, OBL-21, were examined, with an emphasis placed on the role of the microtubular network. We show here that JHMV spread within the central nervous system occurs transneuronally and that virus protein trafficking was dependent upon microtubules. Viral trafficking occurred asymmetrically, involving both the somatodendritic and the axonal domains. Thus coronavirus can be disseminated from neurons at either the basolateral or the apical domains. A specific interaction between antibodies derived against the microtubule-associated protein tau and JHMV nucleocapsid protein (N) was observed, which can presumably be explained by an overall amino acid similarity of 44% and an identity of 20% between proteins N and tau, with optimal alignment at the microtubule binding domain of tau. Collectively, our data suggest an important role of the microtubule network in viral protein trafficking and distribution. They also draw attention to protein sequence mimicry of a cell component by this coronavirus as one strategy for making use of the host's functions on behalf of the virus.


title: A nucleolar protein ApLLP induces ApC/EBP expression required for long-term synaptic facilitation in aplysia neurons.

text: In Aplysia, long-term synaptic plasticity is induced by serotonin (5-HT) or neural activity and requires gene expression. Here, we demonstrate that ApLLP, a novel nucleolus protein, is critically involved in both long-term facilitation (LTF) and behavioral sensitization. Membrane depolarization induced ApLLP expression, which activated ApC/EBP expression through a direct binding to CRE. LTF was produced by a single pulse of 5-HT 30 min after the membrane depolarization. This LTF was blocked when either ApLLP or ApC/EBP were blocked by specific antibodies. In contrast, ApLLP overexpression induced LTF in response to a single 5-HT treatment. Simultaneously, a siphon noxious stimulus (SNS) to intact Aplysia induced ApLLP and ApC/EBP expression, and single tail shock 30 min after SNS transformed short-term sensitization to long-term sensitization of siphon withdrawal reflex. These results suggest that ApLLP is an activity-dependent transcriptional activator that switches short-term facilitation to long-term facilitation.


title: A comprehensive systematic review of healthcare workers' perceptions of risk from exposure to emerging acute respiratory infectious diseases and the perceived effectiveness of strategies used to facilitate healthy coping in acute hospital and community healthcare settings.

text: BACKGROUND Emerging acute respiratory infectious diseases, such as Severe Acute Respiratory Syndrome (SARS) and the Avian Influenza A/H5N1 virus are contagious with high morbidity and mortality rates. Hence, health care workers, who are in close contact with affected patients, face many risks. There need to be a greater understanding of: individual HCWs' risk perceptions; adopted risk-mitigating strategies; and factors influencing both. OBJECTIVES This review aimed to establish the best evidence regarding health care worker's risk perceptions and workplace strategies towards emerging acute respiratory infectious diseases in acute hospital and community healthcare settings; and to make recommendations for practice and future research. INCLUSION CRITERIA Participants Studies that included male and female health care workers practising in acute and community health care settings were considered.Types of intervention (s)/Phenomena of interest This review considered studies that investigated: health care workers' risk perceptions; perceived meaning/effectiveness of the individual and workplace strategies implemented; and the factors influencing both.Types of outcomes This review focused on factors affecting: health care worker' risk perceptions; use of risk-mitigating strategies; and their perceived meaning and effectiveness.Types of studies Both qualitative and quantitative study designs published in the English language were including in the study. SEARCH STRATEGY Using a three-step search strategy, the following databases from 1997-2009 were accessed: CINAHL, PubMed, SCOPUS, ScienceDirect, Sociological Abstracts, PsycINFO, and Web of Science. METHODOLOGICAL QUALITY Two independent reviewers assessed each paper for methodological validity prior to inclusion in the review using standardised critical appraisal instruments from the Joanna Briggs Institute. Disagreements were resolved through discussion, or with a third reviewer. DATA EXTRACTION Information was extracted by two independent reviewers from each paper using the standardised data extraction tools from JBI. DATA SYNTHESIS The findings are presented in narrative form, as textual pooling and statistical pooling was not possible for the qualitative and quantitative papers respectively. RESULTS Ten quantitative studies and one qualitative study were included in the review. Findings indicated that although health care workers perceived personal and familial health risks and stigmatization from their exposure to emerging acute respiratory infectious diseases, the majority were accepting of these risks. Organizational implementation of infection control measures; avoidance of patients; and complying with personal protective equipment were identified as risk-mitigating strategies. Demographic, individual and organizational factors were found to influence their risk perceptions and their adoption of these strategies. CONCLUSIONS There is a nebulous relationship between health care workers' risk perceptions and how these perceptions affect their use of risk-mitigating strategies. Hence, future research needs to examine the nuances of this relationship, as well as the influencing factors involved. In this review, several key factors influencing health care workers' risk perceptions and risk-mitigating strategies were identified and these may form the implications for new practice. IMPLICATIONS FOR PRACTICE Institutions need to: empower health care workers through education and training; protect them with organizational safeguards; and offer incentives to encourage willingness to work, especially for those with high risk perceptions. IMPLICATIONS FOR RESEARCH Future research need to examine the: differences between HCWs' intentions to care during SARS and influenza outbreaks; relationship between intentions to care and actual care provided; difference between their intentions to care/resign and acceptance of risks; factors influencing their intentions to care; and factors influencing compliance to infection control measures.


title: Mutual viral and bacterial infections after housing rats of various breeders within an experimental unit.

text: Fifteen athymic rat strains from 11 breeding colonies were housed within an experimental facility for an immunological study. Health status records supplied with 14 of the strains listed infections by Kilham's rat virus (KRV), Clostridium piliforme (Bacillus piliformis) and Pasteurella pneumotropica for 2, 2 and 1 colonies respectively. In sera taken previous to the study from euthymic rats of 10 strains, antibodies to KRV were detected in 3 strains, to Pneumonia virus of mice (PVM), Rat corona virus (RCV) and Sendai virus in one strain each and to P. pneumotropica in 2 strains. Only 2 of the KRV infections had been reported by the supplier. During the study rats of all 10 strains developed antibodies to 2-4 of viral antigens. Eight out of 10 rat strains seroconverted to 1-5 of the antigens C. piliforme (B. piliformis), Bordetella bronchiseptica, Haemophilus spp., P. pneumotropica and Streptobacillus moniliformis. Two rat strains housed in filtertop cages did not develop antibodies to bacterial antigens. The potential detrimental effects of intercurrent infections on the outcome of the comparative immunological study are discussed.


Taiwan proteomics society: an organization affiliated with HUPO.


title: Recent patents on treatment of severe acute respiratory syndrome (SARS).

text: Severe acute respiratory syndrome (SARS) is an epidemic that spread worldwide in early 2003. The aetiological agent was originally defined as a novel coronavirus and later designated as the SARS coronavirus (SARS-CoV), which appears similar to other coronaviruses in both virion structure and genome organization with a single-stranded, plus-sense RNA. However, the epidemiology and pathogenesis of SARS remain poorly understood and there is currently no effective treatment. To date, considerable research has been done on detection, prevention and treatment of SARS. In this review, we mainly focus on the recent patents and research work on detecting, preventing and treating SARS.


title: Glycyrrhizic acid alters Kaposi sarcoma-associated herpesvirus latency, triggering p53-mediated apoptosis in transformed B lymphocytes.

text: Kaposi sarcoma-associated herpesvirus (KSHV) is linked with all clinical forms of Kaposi sarcoma and several lymphoproliferative disorders. Like other herpesviruses, KSHV becomes latent in the infected cells, expressing only a few genes that are essential for the establishment and maintenance of its latency and for the survival of the infected cells. Inhibiting the expression of these latent genes should lead to eradication of herpesvirus infection. All currently available drugs are ineffective against latent infection. Here we show, for the first time to our knowledge, that latent infection with KSHV in B lymphocytes can be terminated by glycyrrhizic acid (GA), a triterpenoid compound earlier shown to inhibit the lytic replication of other herpesviruses. We demonstrate that GA disrupts latent KSHV infection by downregulating the expression of latency-associated nuclear antigen (LANA) and upregulating the expression of viral cyclin and selectively induces cell death of KSHV-infected cells. We show that reduced levels of LANA lead to p53 reactivation, an increase in ROS, and mitochondrial dysfunction, which result in G1 cell cycle arrest, DNA fragmentation, and oxidative stress-mediated apoptosis. Latent genes are involved in KSHV-induced oncogenesis, and strategies to interfere with their expression might prove useful for eradicating latent KSHV infection and have future therapeutic implications.


title: Regulatory and microbiological safety issues surrounding cell and tissue-engineering products.

text: Cell therapies and tissue-engineered products that contain living cells are potentially some of the most exciting of the novel therapeutic products currently under development. These products, however, present a number of important safety issues, particularly with respect to the transmission of human viruses. In addition, the short shelf life of these products precludes the normally extensive characterization performed on other biotherapeutic products. Careful examination of the risks and extensive testing of the raw materials have been used in place of product testing to ensure safety.


title: Prognostic Factors for Adverse Outcomes in COVID-19 Infection.

text: Whilst COVID-19 infection generally run a mild course in up to 80% of those affected, a number of pre-existing co-morbidities determine the severity of infection and the outcome in an individual patient. The most important of these co-morbidities that have consistently emerged in studies from across the globe, are the patients age and sex. Other important co-morbidities that adversely affect outcomes include pre-existing diabetes, obesity, hypertension, chronic lung disease and malignancy. This comprehensive review discusses the impact of these co-morbidities and the role of laboratory predictors of poor patient outcomes.


title: Human coronavirus 229E papain-like proteases have overlapping specificities but distinct functions in viral replication.

text: Expression of the exceptionally large RNA genomes of CoVs involves multiple regulatory mechanisms, including extensive proteolytic processing of the large replicase polyproteins, pp1a and pp1ab, by two types of cysteine proteases: the chymotrypsin-like main protease and papain-like accessory proteases (PLpros). Here, we characterized the proteolytic processing of the human coronavirus 229E (HCoV-229E) amino-proximal pp1a/pp1ab region by two paralogous PLpro activities. Reverse-genetics data revealed that replacement of the PL2pro active-site cysteine was lethal. By contrast, the PL1pro activity proved to be dispensable for HCoV-229E virus replication, although reversion of the PL1pro active-site substitution to the wild-type sequence after several passages in cell culture indicated that there was selection pressure to restore the PL1pro activity. Further experiments showed that both PL1pro and PL2pro were able to cleave the nsp1-nsp2 cleavage site, with PL2pro cleaving the site less efficiently. The PL1pro-negative mutant genotype could be stably maintained in cell culture when the nsp1-nsp2 site was replaced by a short autoproteolytic sequence, suggesting that the major driving force for the observed reversion of the PL1pro mutation was the requirement for efficient nsp1-nsp2 cleavage. The data suggest that the two HCoV-229E PLpro paralogs have overlapping substrate specificities but different functions in viral replication. Within the tightly controlled interplay of the two protease activities, PL2pro plays a universal and essential proteolytic role that appears to be assisted by the PL1pro paralog at specific sites. Functional and evolutionary implications of the differential amino-terminal polyprotein-processing pathways among the main CoV lineages are discussed.


title: En bloc ligation of renal vessels is safe and reduces duration of surgery.

text: INTRODUCTION Conventionally, individual ligation of the renal vessels with clips is performed during laparoscopic nephrectomy (LN). Concomitant ligation of the vessels is not a standard procedure due to an expected risk of stapler dysfunction and the development of arteriovenous fistulas (AVF). Using the EndoGIA stapler 45/2.5 mm, we compared en bloc ligation with individual ligation during LN and nephroureterectomy (LNU) with a special focus on the development of AVF and technique safety. METHODS This was a retrospective study of all patients undergoing LN or LNU at the Department of Urology, Roskilde Hospital, Denmark, between January 2010 and April 2014. The follow-up period was minimum six months. RESULTS A total of 228 patients underwent LN and 56 patients underwent LNU. In the LN group, 77 patients underwent en bloc ligation. The mean surgical time was significantly reduced to 89 minutes in the en bloc group compared to 109 minutes in the conventional group (p = 0.0001). The difference remained significant with multivariate analysis. In the LNU group, seven patients underwent en bloc ligation. There was no significant difference between conventional ligation and en bloc ligation with respect to surgical time in either the univariate or the multivariate analyses. None of the patients needed blood transfusion. With a mean follow-up of 13.5 months, no AVF were found. CONCLUSIONS En bloc ligation appears to be safe and can reduce the surgical time during LN without increased risk of blood transfusion and without development of AVF. Further studies are needed to assess any advantages associated with use of the method during LNU. FUNDING none. TRIAL REGISTRATION not relevant.


A Potential Complication of Barbed Sutures Preventing Foreign Body Granulomas Induced by Surgical Cloth Particles.


title: cDNA probes for the diagnosis of bovine torovirus (Breda virus) infection.

text: A genomic cDNA library of RNA from Breda virus (BRV), a bovine torovirus, was prepared. The nucleotide sequence of the 3' end of the genome was found to be highly conserved (93% identical) between BRV and Berne virus, the torovirus prototype. Cross-hybridization experiments were performed to select Berne virus cDNA clones for use as probes in a dot hybridization assay; the objective was to detect heterologous torovirus RNA in fecal material. A rapid RNA extraction method was employed to make the test applicable for routine diagnosis. Samples from calves after experimental and natural infection with BRV were assayed to establish the sensitivity and specificity of the test and to compare the test with the enzyme-linked immunosorbent assay (ELISA) for antigen detection. For this purpose, 53 samples from seven infected calves were tested with both methods. In the ELISA, BRV was detected in six fecal samples from three inoculated calves. By use of the hybridization test, 16 samples from seven calves reacted positively. With one exception, only postinoculation samples were found positive in hybridization. No signal was seen in feces from uninoculated calves or from calves infected with rotavirus or coronavirus.


Experimental infection of swine with a sandfly (Lutzomyia shannoni) isolate of vesicular stomatitis virus, New Jersey serotype.


title: Clinical and radiological outcome following pneumothorax after endoscopic lung volume reduction with valves.

text: INTRODUCTION Valve implantation has evolved as a therapy for patients with advanced emphysema. Although it is a minimally invasive treatment, it is associated with complications, the most common being pneumothorax. Pneumothorax occurs due to the rapid target lobe volume reduction and may be a predictor of clinical benefit despite this complication. OBJECTIVE The objective of this study was to conduct an exploratory data analysis of patients who developed a pneumothorax following endoscopic valve therapy for emphysema. MATERIALS AND METHODS This study performed a retrospective evaluation of pneumothorax management and the impact of pneumothorax on clinical outcomes in 70 patients following valve therapy in 381 consecutive patients. RESULTS Pneumothorax rate following valve therapy was 18%. Pneumothorax management consisted of chest tube insertion, valve removal, and surgical intervention in 87% (61/70), 44% (31/70), and 19% (13/70) of the patients, respectively. Despite pneumothorax, patients experienced modest but significant improvements in lung function parameters (forced expiratory volume in 1 second: 55±148 mL, residual volume: -390±964 mL, total lung capacity: -348±876; all P<0.05). Persistent lobar atelectasis 3 months after recovering from pneumothorax, which was associated with relevant clinical improvement, was observed in only 21% (15/70) of the patients. CONCLUSION Pneumothorax is a frequent severe complication following valve therapy that requires further intervention. Nevertheless, the pneumothorax does not impair the clinical status in the majority of patients. Patients with lobar atelectasis benefit after recovering from pneumothorax in terms of lung function parameters.


title: Augmentation of influenza surveillance with rapid antigen detection at the point-of-care: results of a pilot study in Tasmania, 2004.

text: Tasmania contributes very few laboratory confirmed cases to Australia's national influenza surveillance statistics. In 2004, a study was conducted to pilot test sentinel syndromic surveillance for influenza-like illness supplemented by point-of-care testing using the Binax Now Flu A Test Kit and by viral culture, to assess the feasibility and acceptability of this method of surveillance. Overall, the goal of such a system would be to increase laboratory surveillance activity within Tasmania and increase the number of specimens sent for viral culture. Five sites participated in the study, including three public hospital emergency departments and two general practices. Despite being conducted during a period of low influenza activity, the pilot study demonstrated that augmentation of syndromic surveillance with point-of-care testing is both feasible and acceptable but is best conducted in the general practice setting.


title: May titanium spiral tacks contribute to intra-abdominal adhesion formation?

text: PURPOSE To verify if titanium spiral tacks may contribute to intra-abdominal adhesion formation in rabbits. METHODS Ten New Zealand white rabbits were used. Through videolaparoscopic surgery two titanium spiral tacks (ProTack(®), Covidien(®)) were applied in the right flank. After 30 days the animals were submitted to evaluate peritoneal cavity by videolaparoscopy. RESULTS No adhesion was found where the titanium spiral tacks were placed. CONCLUSION Titanium spiral tacks for fixation do not cause adhesions in rabbits.


title: A primer of primate pathology: lesions and nonlesions.

text: Nonhuman primates are important laboratory animals for biomedical, pharmacology, and toxicology research. To effectively use primates as models, their gross and histologic anatomy, physiology and natural history, as well as common health problems and the source from which the primate is obtained, must be known and understood by pathologists involved in study design and/or interpretation. The first very important lesson in the "primer" is: there is no such thing as a generic monkey. Brand names (ie, species and subspecies) are important. Several taxonomic groups of primates are used in research including: prosimians, such as galagos and lemurs; New World monkeys, particularily marmosets; Old World monkeys, especially macaques and baboons; and the chimpanzee, an African ape. Differences between taxa are exemplified by the glucocorticoid resistance of New World monkeys compared to Old World monkeys, which results in the requirement for Vitamin D3 and their high circulating levels of steroids such as cortisone and progesterone. Differences in ovarian histology between Old and New World monkeys probably relate to steroid receptor biology as well. There are also variations in disease manifestations, even among closely related primate species such as rhesus and cynomolgus macaques (cynos). For example type D retrovirus infection is accompanied by lymphomas in cynos, but not rhesus. The second important lesson in this "primer" is: "not test article related" does not always mean "normal." Lymphoid nodules in bone marrow or salivary gland, a common background finding in macaques, often signal the presence of type D retrovirus. Other histologic changes and normal anatomic variations may be confusing to individuals not routinely examining primate tissues. The objective of this paper is to familiarize pathologists with the use of primates in research as well as lesions and nonlesions (normal anatomy or physiology) of primates that may influence study design and confound interpretation.


title: Emergency medical services.

text: Prehospital emergency medical service personnel are at occupational risk of exposure to human immunodeficiency virus (HIV). Current protections from blood-borne pathogens are not adequate for all firefighters and emergency medical services (EMS) personnel. The International Association of Fire Fighters examines the environmental realities of the profession and seeks solutions for effective postexposure management of HIV-exposed firefighters and EMS professionals.


title: Occurrence and properties of FY(Att25)+ Escherichia coli associated with diarrhoea in calves.

text: Escherichia coli producing the adhesive antigen FY(Att25) were isolated from 46 of 1341 (3.4 per cent) E coli isolated from calves on 20 of 164 (12.1 per cent) farms in Scotland and England. Twenty of the 46 calves had diarrhoea and in nine of these animals there were mixed infections with rotavirus, coronavirus, cryptosporidium and Salmonella typhimurium. The F41 fimbrial adhesin was found on one of the FY(Att25)+ E coli. This strain also produced heat stable enterotoxin. The remaining FY(Att25)+ isolates did not produce other adhesins, enterotoxins or verotoxins. The FY(Att25) antigen was not detected on 109 pathogenic E coli isolated from calves, chickens, lambs, pigs or man.


Putting SARS in perspective: a communication challenge.


title: Recognizing and managing emerging infectious diseases in the emergency department [digest].

text: With population shifts, increased travel, and climate change, the spread of emerging and re-emerging infections is increasing. Although encountering a patient with an emerging infection on any given emergency department shift is unlikely, missing a diagnosis could have profound consequences for the patient, healthcare workers, and the patient's close contacts. This review provides a framework to evaluate, diagnose, and treat a returning traveler with suspected Middle East respiratory syndrome, chikungunya virus, or Zika virus-3 recently emerged infections. All may present with nonspecific viral-like symptoms and are easily missed if an appropriate travel history is not obtained. A high level of vigilance and proper disposition will enable the emergency clinician to effectively diagnose, manage, and contain these diseases. [Points & Pearls is a digest of Emergency Medicine Practice.].


title: Comparative analysis of 22 coronavirus HKU1 genomes reveals a novel genotype and evidence of natural recombination in coronavirus HKU1.

text: We sequenced and compared the complete genomes of 22 strains of coronavirus HKU1 (CoV HKU1) obtained from nasopharyngeal aspirates of patients with respiratory tract infections over a 2-year period. Phylogenetic analysis of 24 putative proteins and polypeptides showed that the 22 CoV HKU1 strains fell into three clusters (genotype A, 13 strains; genotype B, 3 strains and genotype C, 6 strains). However, different phylogenetic relationships among the three clusters were observed in different regions of their genomes. From nsp4 to nsp6, the genotype A strains were clustered with the genotype B strains. For nsp7 and nsp8 and from nsp10 to nsp16, the genotype A strains were clustered with the genotype C strains. From hemagglutinin esterase (HE) to nucleocapsid (N), the genotype B strains were clustered closely with the genotype C strains. Bootscan analysis showed possible recombination between genotypes B and C from nucleotide positions 11,500 to 13,000, corresponding to the nsp6-nsp7 junction, giving rise to genotype A, and between genotypes A and B from nucleotide positions 21,500 to 22,500, corresponding to the nsp16-HE junction, giving rise to genotype C. Multiple alignments further narrowed the sites of crossover to a 143-bp region between nucleotide positions 11,750 and 11,892 and a 29-bp region between nucleotide positions 21,502 and 21,530. Genome analysis also revealed various numbers of tandem copies of a perfect 30-base acidic tandem repeat (ATR) which encodes NDDEDVVTGD and various numbers and sequences of imperfect repeats in the N terminus of nsp3 inside the acidic domain upstream of papain-like protease 1 among the 22 genomes. All 10 CoV HKU1 strains with incomplete imperfect repeats (1.4 and 4.4) belonged to genotype A. The present study represents the first evidence for natural recombination in coronavirus associated with human infection. Analysis of a single gene is not sufficient for the genotyping of CoV HKU1 strains but requires amplification and sequencing of at least two gene loci, one from nsp10 to nsp16 (e.g., pol or helicase) and another from HE to N (e.g., spike or N). Further studies will delineate whether the ATR is useful for the molecular typing of CoV HKU1.


title: A reversibly palmitoylated resident protein (p63) of an ER-Golgi intermediate compartment is related to a circulatory shock resuscitation protein.

text: The recently identified 63 kDa membrane protein, p63, is a resident protein of a membrane network interposed in between rough ER and Golgi apparatus. To characterize p63 at the molecular level a 2.91 kb cDNA encoding p63 has been isolated from a human placenta lambda gt10 cDNA library. Sequence analysis of tryptic peptides prepared from isolated p63 confirmed the identify of the cloned gene. The translated amino acid sequence consists of 601 amino acids (65.8 kDa) with a single putative membrane-spanning region and a N-terminal cytoplasmic domain of 106 amino acids. The human p63 cDNA exhibits a high level of sequence identify to the pig hepatic cDNA 3AL (accession number M27092) whose expression is enhanced after resuscitation from circulatory shock. An additional remarkable feature of p63 is that it becomes reversibly palmitoylated when intracellular protein transport is blocked by the drug brefeldin A. Overexpression of p63 in COS cells led to the development of a striking tubular membrane network in the cytoplasm. This suggests that the protein may be determinant for the structure of the p63 compartment.


Rotavirus and coronavirus associated diarrohoea in calves.


title: The C-type lectin L-SIGN differentially recognizes glycan antigens on egg glycosphingolipids and soluble egg glycoproteins from Schistosoma mansoni.

text: Recognition of pathogen-derived carbohydrate constituents by antigen presenting cells is an important step in the induction of protective immunity. Here we investigated the interaction of L-SIGN (liver/lymph node specific ICAM-3-grabbing nonintegrin), a C-type lectin that functions as antigen receptor on human liver sinusoidal endothelial cells, with egg-derived glycan antigens of the parasitic trematode Schistosoma mansoni. Our data demonstrate that L-SIGN binds both schistosomal soluble egg antigens (SEA) and egg glycosphingolipids, and can mediate internalization of SEA by L-SIGN expressing cells. Binding and internalization of SEA was strongly reduced after treatment of SEA with endoglycosidase H, whereas defucosylation affected neither binding nor internalization. These data indicate that L-SIGN predominantly interacts with oligomannosidic N-glycans of SEA. In contrast, binding to egg glycosphingolipids was completely abolished after defucosylation. Our data show that L-SIGN binds to a glycosphingolipid fraction containing fucosylated species with compositions of Hex(1)HexNAc(5-7)dHex(3-6)Cer, as evidenced by mass spectrometry. The L-SIGN "gain of function" mutant Ser363Val, which binds fucosylated Lewis antigens, did not bind to this fucosylated egg glycosphingolipid fraction, suggesting that L-SIGN displays different modes in binding fucoses of egg glycosphingolipids and Lewis antigens, respectively. Molecular modeling studies indicate that the preferred binding mode of L-SIGN to the respective fucosylated egg glycosphingolipid oligosaccharides involves a Fucalpha1-3GalNAcbeta1-4(Fucalpha1-3)GlcNAc tetrasaccharide at the nonreducing end. In conclusion, our data indicate that L-SIGN recognizes both oligomannosidic N-glycans and multiply fucosylated carbohydrate motifs within Schistosoma egg antigens, which demonstrates that L-SIGN has a broad but specific glycan recognition profile.


title: Coronavirus proteins: biogenesis of avian infectious bronchitis virus virion proteins.

text: We examined the synthesis of viral structural proteins in cultured cells infected with the avian coronavirus infectious bronchitis virus. Tryptic peptide mapping was used to determine the structural relationships of the intracellular proteins to the virion polypeptides. Pulse-chase experiments were performed to identify precursors to the virus-specific proteins. We found that the nucleocapsid protein, P51, and the small viral membrane proteins GP31, GP28, and P23 do not undergo post-translational proteolytic processing. In contrast, GP90 and GP84, the two large virion membrane proteins, were found to be produced by cleavage of a single precursor, GP155. This demonstrated that at least one coronavirus mRNA specifies two virion proteins.


title: Structural basis for coronavirus-mediated membrane fusion. Crystal structure of mouse hepatitis virus spike protein fusion core.

text: The surface transmembrane glycoprotein is responsible for mediating virion attachment to cell and subsequent virus-cell membrane fusion. However, the molecular mechanisms for the viral entry of coronaviruses remain poorly understood. The crystal structure of the fusion core of mouse hepatitis virus S protein, which represents the first fusion core structure of any coronavirus, reveals a central hydrophobic coiled coil trimer surrounded by three helices in an oblique, antiparallel manner. This structure shares significant similarity with both the low pH-induced conformation of influenza hemagglutinin and fusion core of HIV gp41, indicating that the structure represents a fusion-active state formed after several conformational changes. Our results also indicate that the mechanisms for the viral fusion of coronaviruses are similar to those of influenza virus and HIV. The coiled coil structure has unique features, which are different from other viral fusion cores. Highly conserved heptad repeat 1 (HR1) and HR2 regions in coronavirus spike proteins indicate a similar three-dimensional structure among these fusion cores and common mechanisms for the viral fusion. We have proposed the binding regions of HR1 and HR2 of other coronaviruses and a structure model of their fusion core based on our mouse hepatitis virus fusion core structure and sequence alignment. Drug discovery strategies aimed at inhibiting viral entry by blocking hairpin formation may be applied to the inhibition of a number of emerging infectious diseases, including severe acute respiratory syndrome.


Full-Thickness Interrupted Ligation with Transhepatic Suturing (the FILTH Technique): A Novel Technique for Hepatic Trauma.


title: [Diagnosis and treatment recommendation for pediatric COVID-19 (the second edition)].

text: The coronavirus disease 2019 (COVID-19) has caused a global pandemic. All people including children are generally susceptible to COVID-19, but the condition is relatively mild for children. The diagnosis of COVID-19 is largely based on the epidemiological evidence and clinical manifestations, and confirmed by positive detection of virus nucleic acid in respiratory samples. The main symptoms of COVID-19 in children are fever and cough; the total number of white blood cell count is usually normal or decreased; the chest imaging is characterized by interstitial pneumonia, which is similar to other respiratory virus infections and Mycoplasma pneumoniae infections. Early identification, early isolation, early diagnosis and early treatment are important for clinical management. The treatment of mild or moderate type of child COVID-19 is mainly symptomatic. For severe and critical ill cases, the oxygen therapy, antiviral drugs, antibacterial drugs, glucocorticoids, mechanical ventilation or even extracorporeal membrane oxygenation (ECMO) may be adopted, and the treatment plan should be adjusted timely through multi-disciplinary cooperation.


Synchronized cardioversion of unstable supraventricular tachycardia resulting in ventricular fibrillation.


title: Analysis of the two subgenomic RNA promoters for turnip crinkle virus in vivo and in vitro.

text: Infection of plants or protoplasts with turnip crinkle virus (TCV), a monopartite RNA virus, results in the synthesis of the genomic RNA and two subgenomic (sg) RNAs. The transcription start site for the 1.45-kb sgRNA was previously mapped to position 2606 (J. C. Carrington, T. J. Morris, P. G. Stockley, and S. C. Harrison, (1987). J. Mol. Biol. 194, 265-276) corresponding to position 2607 in the TCVms isolate and the start site for the 1.7-kb sgRNA has now been mapped to position 2333 in TCVms. A 96-base sequence (90 bases upstream and 6 bases downstream) encompassing the transcription start site for the 1.45-kb sgRNA was sufficient for full promoter activity. Similarly, a 94-base sequence (90 bases upstream and 4 bases downstream) encompassing the start site was required for full activity of the 1.7-kb sgRNA promoter. The 1.45-kb sgRNA promoter, but not the 1.7-kb sgRNA promoter, was able to direct synthesis of a nontemplate RNA in vitro using partially purified TCV RNA-dependent RNA polymerase. Computer generated secondary structures for the two sgRNA promoters revealed an extensive hairpin just upstream from the transcription start site. Comparisons of corresponding sequences from related viruses indicates higher sequence conservation for the 1.45-kb sgRNA promoter compared with the 1.7-kb sgRNA promoter, despite the latter's location within the RNA-dependent RNA polymerase open reading frame.


title: Conserved leucines in N-terminal heptad repeat HR1 of envelope fusion protein F of group II nucleopolyhedroviruses are important for correct processing and essential for fusogenicity.

text: The heptad repeat (HR), a conserved structural motif of class I viral fusion proteins, is responsible for the formation of a six-helix bundle structure during the envelope fusion process. The insect baculovirus F protein is a newly found budded virus envelope fusion protein which possesses common features to class I fusion proteins, such as proteolytic cleavage and the presence of an N-terminal open fusion peptide and multiple HR domains on the transmembrane subunit F(1). Similar to many vertebrate viral fusion proteins, a conserved leucine zipper motif is predicted in this HR region proximal to the fusion peptide in baculovirus F proteins. To facilitate our understanding of the functional role of this leucine zipper-like HR1 domain in baculovirus F protein synthesis, processing, and viral infectivity, key leucine residues (Leu209, Leu216, and Leu223) were replaced by alanine (A) or arginine (R), respectively. By using Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) as a pseudotype expression system, we demonstrated that all mutant F proteins incorporated into budded virus, indicating that leucine substitutions did not affect intercellular trafficking of F. Furin-like protease cleavage was not affected by any of the leucine substitutions; however, the disulfide bridging and N-linked glycosylation patterns were partly altered. Single substitutions in HR1 showed that the three leucine residues were critical for F fusogenicity and the rescue of AcMNPV infectivity. Our results support the view that the leucine zipper-like HR1 domain is important to safeguard the proper folding, glycosylation, and fusogenicity of baculovirus F proteins.


title: Annual influenza vaccination in community-dwelling elderly individuals and the risk of lower respiratory tract infections or pneumonia.

text: BACKGROUND Influenza vaccination has been associated with a reduction in the number of hospitalizations for respiratory conditions in elderly persons over the period from 1996 to 2002. Little is known, however, about the effect of influenza vaccination on the whole range of severity of respiratory tract infections. METHODS We investigated the effect of annual influenza vaccination on the occurrence of lower respiratory tract infections (LRTIs) in community-dwelling elderly individuals. From 1996 to 2002, we performed a population-based cohort study, using the computerized Integrated Primary Care Information database in the Netherlands, of community-dwelling subjects who were 65 years or older on January 1 of the year of study entry. For each year, the individual cumulative exposure to influenza vaccination since study entry was computed. We compared the risk of LRTI after a first vaccination or revaccination with the risk for no vaccination using a time-varying multivariate Cox proportional hazard model, adjusted for age, sex, smoking, and underlying disease. RESULTS In the study population of 26 071 subjects, 3412 developed LRTIs during follow-up. During the influenza epidemic periods, a first vaccination did not reduce risk for LRTI. In the total population, the hazard ratio following a first vaccination was 0.86 (95% confidence interval [CI], 0.71 to 1.05); in the population without or with comorbidity, these ratios were 0.90 (95% CI, 0.56 to 1.45) and 0.83 (95% CI, 0.66 to 1.04), respectively. During epidemic periods, revaccination reduced risk of LRTI by 33% (95% CI, 8% to 52%) in individuals without comorbidity. In individuals with comorbidity, the risk reduction of 5% was nonsignificant (95% CI, -10% to 18%). CONCLUSION In this study, annual influenza revaccination was associated with a reduction in LRTI in community-dwelling elderly individuals.


Complete genomic sequence of the transmissible gastroenteritis virus.


title: [Characterization and development of recombinant vaccinia viruses expressing different segments of spike protein derived from human coronavirus NL-63].

text: The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.


title: Unruptured internal carotid artery bifurcation aneurysms: general features and overall results after modern treatment.

text: BACKGROUND Internal carotid artery bifurcation aneurysms (ICAbifAs) present unique challenges to endovascular and surgical operators, and little is known about their natural history. We reviewed our institution's experience with ICAbifAs studying outcomes of surgical and endovascular management and natural history. METHODS Consecutive patients with unruptured ICAbifAs evaluated and/or treated over an 8-year interval were studied. Baseline demographics, neurovascular risk factors, aneurysm location and size, clinical presentation, treatment recommendations, and outcomes were prospectively collected and retrospectively analyzed. Continuous variables were compared with Student's t test and categorical variables with Chi-square tests. RESULTS Fifty-nine patients with 61 unruptured ICAbifAs were included. Seven aneurysms were treated surgically (11.5 %), 22 underwent endovascular treatment (36 %), and 32 were managed conservatively (52.5 %). In the surgical group, short- and long-term complete aneurysm occlusion rates were 100 % with no cases of perioperative or long-term permanent morbidity or treatment-related mortality. In the endovascular group, two patients (11.7 %) with giant aneurysms had perioperative thromboembolic events with transient morbidity. There was one case of aneurysm rupture at follow-up in a giant aneurysm treated with partial coil embolization. Complete/near-complete occlusion rates were 63 %. There was one case of aneurysm rupture after 114 aneurysm-years of follow-up in the conservative management group (0.89 %/year), but no ruptures were observed in small aneurysms selected for conservative management. CONCLUSIONS Unruptured small ICAbifAs have a benign natural history. In patients selected for treatment, excellent results can be achieved in the vast majority of patients with judicious use of endovascular and surgical therapy.


title: Bovine-murine hybridoma that secretes bovine monoclonal antibody of defined specificity.

text: Spleen cells from a calf hyperimmunized with bovine enteric coronavirus were fused with nonproducer mouse plasmacytoma cells. Stable hybridoma lines secreting bovine immunoglobulins were obtained. One line secreted monoclonal bovine immunoglobulin G2, which reacted specifically with bovine enteric coronavirus in an enzyme-linked immunosorbent assay, inhibited virus hemagglutination, and precipitated a structural polypeptide with a molecular weight of 26,000 daltons.


title: Systems biology: where it's at in 2005.

text: A report on the joint Keystone Symposia on Systems and Biology and Proteomics and Bioinformatics, Keystone, USA, 8-13 April 2005.


title: Digestive system manifestations in patients with severe acute respiratory syndrome.

text: OBJECTIVE To explore digestive system manifestations in patients with severe acute respiratory syndrome (SARS). METHOD The clinical data of 96 cases with SARS admitted into our hospital from February 6, 2003 to March 28, 2003 were retrospectively analyzed. RESULTS Among the 96 cases, 26 cases (27%) had diarrhea, 17 (18%) had nausea, 6 (6%) had vomiting, 16 (17%) had bellyache, and 8 (8%) had ALT elevation. CONCLUSIONS Patients with SARS may have digestive system manifestations; diarrhea is the most common symptom.


title: A cluster of cases of severe acute respiratory syndrome in Hong Kong.

text: BACKGROUND Information on the clinical features of the severe acute respiratory syndrome (SARS) will be of value to physicians caring for patients suspected of having this disorder. METHODS We abstracted data on the clinical presentation and course of disease in 10 epidemiologically linked Chinese patients (5 men and 5 women 38 to 72 years old) in whom SARS was diagnosed between February 22, 2003, and March 22, 2003, at our hospitals in Hong Kong, China. RESULTS Exposure between the source patient and subsequent patients ranged from minimal to that between patient and health care provider. The incubation period ranged from 2 to 11 days. All patients presented with fever (temperature, >38 degrees C for over 24 hours), and most presented with rigor, dry cough, dyspnea, malaise, headache, and hypoxemia. Physical examination of the chest revealed crackles and percussion dullness. Lymphopenia was observed in nine patients, and most patients had mildly elevated aminotransferase levels but normal serum creatinine levels. Serial chest radiographs showed progressive air-space disease. Two patients died of progressive respiratory failure; histologic analysis of their lungs showed diffuse alveolar damage. There was no evidence of infection by Mycoplasma pneumoniae, Chlamydia pneumoniae, or Legionella pneumophila. All patients received corticosteroid and ribavirin therapy a mean (+/-SD) of 9.6+/-5.42 days after the onset of symptoms, and eight were treated earlier with a combination of beta-lactams and macrolide for 4+/-1.9 days, with no clinical or radiologic efficacy. CONCLUSIONS SARS appears to be infectious in origin. Fever followed by rapidly progressive respiratory compromise is the key complex of signs and symptoms from which the syndrome derives its name. The microbiologic origins of SARS remain unclear.


The threat of an avian influenza pandemic.


Characterization of viral proteins synthesized in 229E infected cells and effect(s) of inhibition of glycosylation and glycoprotein transport.


title: The rapid pull-back technique for navigation across a wide-necked aneurysm. a report of four cases.

text: Embolization of wide-necked and/or giant aneurysms may fail due to the inability to pass across the aneurysm neck. We describe the rapid bull-back technique used in four patients in which a small diameter microcatheter with the aid of a hydrophilic microguidewire was navigated along the inner surface of the aneurysm, making a loop in the dome, exiting the neck to reach distal intracranial vessels. After withdrawal of microguidewire, the microcatheter is pulled back rapidly up to a predetermined length. This maneuver results in elimination of the loop, straightening the microcatheter to allow an exchange procedure for another device to cross the neck distally and continue the embolization procedure. The rapid pull-back technique is useful during the endovascular treatment of wide-necked and/or giant aneurysms as it helps to achieve reliable access to the distal parent vessel with the microcatheter. This is of increasing importance since an increasing number of aneurysms will be treated in the future with refinements in various intracranial stents.


title: Effect of widespread restrictions on the use of hospital services during an outbreak of severe acute respiratory syndrome.

text: BACKGROUND Restrictions on the nonurgent use of hospital services were imposed in March 2003 to control an outbreak of severe acute respiratory syndrome (SARS) in Toronto, Ont. We describe the impact of these restrictions on health care utilization and suggest lessons for future epidemics. METHODS We performed a retrospective population-based study of the Greater Toronto Area (hereafter referred to as Toronto) and unaffected comparison regions (Ottawa and London, Ont.) before, during and after the SARS outbreak (April 2001-March 2004). We determined the adjusted rates of hospital admissions, emergency department and outpatient visits, diagnostic testing and drug prescribing. RESULTS During the early and late SARS restriction periods, the rate of overall and medical admissions decreased by 10%-12% in Toronto; there was no change in the comparison regions. The rate of elective surgery in Toronto fell by 22% and 15% during the early and late restriction periods respectively and by 8% in the comparison regions. The admission rates for urgent surgery remained unchanged in all regions; those for some acute serious medical conditions decreased by 15%-21%. The rates of elective cardiac procedures declined by up to 66% in Toronto and by 71% in the comparison regions; the rates of urgent and semi-urgent procedures declined little or increased. High-acuity visits to emergency departments fell by 37% in Toronto, and inter-hospital patient transfers fell by 44% in the circum-Toronto area. Drug prescribing and primary care visits were unchanged in all regions. INTERPRETATION The restrictions achieved modest reductions in overall hospital admissions and substantial reductions in the use of elective services. Brief reductions occurred in admissions for some acute serious conditions, high-acuity visits to emergency departments and inter-hospital patient transfers suggesting that access to care for some potentially seriously ill patients was affected.


Adenovirus infection in Spanish ibex.


title: Articulating and Reloadable Fixation Devices for Hernia Repair.

text: There have been a variety of absorbable and permanent tacks, tack deployment systems, and fasteners developed for the fixation of mesh during laparoscopic ventral hernia repair. The manufacturer recommendation for all systems is for perpendicular deployment of these tacks into the tissue. Achieving this optimal angle with previously developed deployment systems is often challenging and can lead to tack failure, mesh migration, and recurrence, or may require the placement of additional ports. Additionally, current tack deployment systems lack the ability to reload, leading to increased cost when entire systems must be opened each time a reload is necessary. This article presents products designed to addresses both of these problems. These deployment instruments incorporate an articulating shaft or a hinge mechanism allowing for improved access to different parts of the abdominal cavity and delivering perpendicular placement of tacks with fewer port sites. Devices with the option of reloadable fixation decrease costs and reduce waste.


In vivo diagnosis of feline infectious peritonitis by comparison of protein content, cytology, and direct immunofluorescence test on peritoneal and pleural effusions.


Molecular biology of coronavirus 1986.


title: Virus-cell interactions in the induction of type 1 interferon by influenza virus in mouse spleen cells.

text: Inactivated influenza A virus and fixed, virus-infected cells induce type 1 interferon (IFN-alpha/beta) production in murine splenocytes. In this study, we have explored the nature of the virus-spleen cell interaction that leads to IFN-alpha/beta induction and the reason for the poor response to some virus strains. IFN-alpha/beta induction by horse serum-sensitive, but not -resistant, strains of influenza virus was inhibited in the presence of horse serum, indicating that binding of the virus to sialylated cell receptors is a necessary step in the induction process. Furthermore, influenza viruses A/PR/8/34 (H1N1) and A/WS/33 (H1N1), which were poor inducers of IFN-alpha/beta in spleen cells, were shown to have a more active neuraminidase than strains that induced higher IFN levels, and IFN-alpha/beta induction by A/PR/8/34 (H1N1) and A/WS/33 (H1N1) was restored in the presence of a neuraminidase inhibitor. Growth of virus in different cell types altered the level of IFN-alpha/beta induced in spleen cells by particular virus strains, suggesting that the nature of the carbohydrate moieties on the viral glycoproteins may also influence IFN-alpha/beta induction in this system. Consistent with this notion, treatment of egg-grown virus with periodate to oxidize viral carbohydrate greatly reduced its capacity for IFN-alpha/beta induction. Furthermore, induction of IFN-alpha/beta was inhibited in the presence of the saccharides yeast mannan and laminarin. Together these findings indicate: (i) a requirement for interaction of the virus with sialylated receptors on the IFN-producing cell; (ii) an influence of viral carbohydrate on the response; and (iii) possible involvement of a lectin-like receptor on the IFN-producing cell in the induction of IFN-alpha/beta or in regulation of this response.


title: Tyzzer's disease in a red panda (Ailurus fulgens fulgens).

text: A debilitated 9-yr-old female red panda (Ailurus fulgens fulgens) with a recent history of corticosteroid administration displayed anorexia, depression, and diarrhea for 2 days. Blood work revealed a moderate nonregenerative anemia, leukocytosis, hypokalemia, hyperbilirubinemia, and mildly elevated alanine aminotransferase and aspartate aminotransferase. Serology was negative for occult heartworm, Toxoplasma gondii, feline leukemia virus, feline infectious peritonitis, feline immunodeficiency virus, and canine distemper virus. Electron microscopy of the feces demonstrated corona-like virus particles. The panda died 3 days after initial presentation. Histologic findings included multifocal, acute, hepatic necrosis and diffuse, necrotizing colitis. Liver and colon lesions contained intracellular, curved, spore-forming, gram-negative, silver-positive rods morphologically consistent with Clostridium piliforme. This panda most likely contracted Tyzzer's disease subsequent to having a compromised immune system after corticosteroid administration and concurrent disease.


title: Emergence of a convex trade-off between transmission and virulence.

text: Most models of virulence evolution assume that a parasite cannot raise its transmission rate without causing more harm to its host. However, the existence of such trade-off relationships has recently been challenged. Here, we study how a trade-off can emerge from a model that explicitly incorporates within-host dynamics. We find that the existence and the convexity of the trade-off are robust, which implies a definite level of evolutionarily stable virulence (ESV) for the parasite. However, we also show that the dependence of the ESV on parameter values may be very strong. One possible consequence of this sensitivity is that relationships between transmission and virulence observed across populations need not conform to the patterns expected on the basis of a common (fixed) trade-off. We discuss possible experiments and implications of our results for the development of virulence management strategies.


title: Surgical management of thoracoabdominal aneurysms.

text: Thoracoabdominal aneurysm of the aorta (TAAA) is a morbid condition, the treatment of which can be associated with high mortality and complication rates, as well as prolonged length of hospital stay. Currently, three approaches to treatment of TAAAs exist: open, endovascular and hybrid repair. Over the past three decades, a significant decrease in postoperative mortality and paraplegia rates has been achieved due to effective application of such treatment adjuncts as left heart bypass, cerebrospinal fluid drainage, application of hypothermia and neuromonitoring, thereby making surgical treatment of TAAAs increasingly safer for the patient. In this report, we review indications and current approaches to surgical management of TAAAs, as well as the factors that influence the efficacy of this type of treatment.


title: Severe respiratory disease in dairy cows caused by infection with bovine respiratory syncytial virus.

text: Outbreaks of severe respiratory disease caused by bovine respiratory syncytial virus (BRSV) were recorded in dairy herds throughout Sweden in 1988 and subsequently. The virus was demonstrated in nasopharyngeal swab material from animals in the acute stage of the disease by culture, the polymerase chain reaction (PCR) and by immunofluorescence. Serological data from the herds investigated showed that the cows had seroconverted to BRSV rather than to bovine coronavirus, bovine viral diarrhoea virus or parainfluenza-3 virus. It was predominantly dairy herds in isolated areas that contracted a severe primary BRSV infection, often after the purchase of new animals. A nationwide survey for BRSV antibodies in bulk milk samples showed the highest prevalence, of 84 to 89 per cent, in the southernmost regions of Sweden and the lowest prevalence, of 41 to 51 per cent, in the north of the country. The prevalence of BRSV was highest in areas with the highest populations of cattle.


title: Effect of integrated traditional Chinese and Western medicine on SARS: a review of clinical evidence.

text: AIM To assess the possible effect of integrated traditional Chinese and Western medicine on severe acute respiratory syndromes. METHODS The current available randomized controlled trials of integrated traditional Chinese and Western medicine on SARS were identified through systematically searching literature in any languages or any types of publications. Additional studies of gray literature were also collected. The quality of studies was evaluated by two investigators independently based largely on the quality criteria specified CONSORT. Statistical analysis of the results was performed using RevMan 4.2.0 software developed by the Cochrane Collaboration. RESULTS Six studies (n = 366) fulfilling the inclusion criteria were found, of which the quality of one study was graded as B, the remaining five were graded as C. Two studies were performed with meta-analysis, the other four studies existed some heterogeneity for which meta-analysis could not be performed, a significant effect on lung infiltrate absorption was found in the treatment groups of these two studies (RR 6.68, 95% CI (2.93, 15.24), P<0.01), there was no significant differences between the mortality (RR 0.86, 95% CI (0.22, 3.29), P = 0.82) and the average dosage of corticosteroid (WMD -39.65, 95% CI (-116.84, 37.54), P = 0.31). The other three studies also showed significant differences in infiltrate absorption, including national drug No. 2. 3. 4 in combination with Western medicine (RR 5.45, 95% CI (1.54, 19.26)), compound formulas NO. 1 combined with Western medicine (WMD 0.24, 95% CI (0.02, 0.46)), compound formulas combined with Western medicine (RR 8.06, 95% CI (0.40, 163.21)). Kangfeidian No.4 in combination with Western medicine had no significant effect on symptom improvement such as loss of dyspnea and cough (RR 1.50, 95%CI (0.41, 5.43)) and (RR 1.29, 95%CI (0.30, 5.43)). CONCLUSION Integrated traditional Chinese and Western medicines has some positive effects on lung infiltrate absorption in SARS patients, and is recommended as an adjunct treatment for SARS. However, its effect on SARS requires further careful study due to limited available randomized control trials.


title: Middle East respiratory syndrome coronavirus (MERS-CoV) infections in two returning travellers in the Netherlands, May 2014.

text: Two patients, returning to the Netherlands from pilgrimage in Medina and Mecca, Kingdom of Saudi Arabia, were diagnosed with Middle East respiratory syndrome coronavirus (MERS-CoV) infection in May 2014. The source and mode of transmission have not yet been determined. Hospital-acquired infection and community-acquired infection are both possible.


title: Virus entry into a polarized epithelial cell line (MDCK): similarities and dissimilarities between influenza C virus and bovine coronavirus.

text: We have analysed the uptake of influenza C virus and bovine coronavirus (BCV) by a polarized epithelial cell line, Madin-Darby canine kidney (MDCK) cells. Both viruses use N-acetyl-9-O-acetylneuraminic acid as a receptor determinant for attachment to cells. Virus binding assays with immobilized proteins indicated that a glycoprotein of 40 kDa is the major surface protein containing the receptor determinant for the two viruses. MDCK cells grown on filters for permeable support were found to have differential sensitivity to infection by these viruses. Both viruses were able to initiate infection via the apical domain of the plasma membrane, but only influenza C virus also accomplished infection via the basolateral plasma membrane. The resistance of MDCK cells to BCV infection from the basal filter chamber was overcome when the cell polarity was abolished by maintaining the cells in calcium-free medium. This finding indicates that the resistance to basolateral infection by BCV is a property of the cell line and not due to a technical problem related to the use of filters. Our results indicate that two viruses which use the same receptor for attachment to cells may differ in their ability to enter polarized cells. The possible involvement of an accessory molecule in the entry of BCV is discussed.


Rover barks before Beagle as Mars success lifts NASA's spirits.


title: Safety and immunogenicity from a phase I trial of inactivated severe acute respiratory syndrome coronavirus vaccine.

text: BACKGROUND Emergence of severe acute respiratory syndrome (SARS) from the winter of 2002 to the spring of 2003 has caused a serious threat to public health. METHODS To evaluate the safety and immunogenicity of the inactivated SARS coronavirus (SARS-CoV) vaccine, 36 subjects received two doses of 16 SARS-CoV units (SU) or 32 SU inactivated SARS-CoV vaccine, or placebo control. RESULTS On day 42, the seroconversion reached 100% for both vaccine groups. On day 56, 100% of participants in the group receiving 16 SU and 91.1% in the group receiving 32 SU had seroconverted. The geometric mean titre of neutralizing antibody peaked 2 weeks after the second vaccination, but decreased 4 weeks later. CONCLUSION The inactivated vaccine was safe and well tolerated and can elicit SARS-CoV-specific neutralizing antibodies.


title: Detection of rhinovirus, respiratory syncytial virus, and coronavirus infections in acute otitis media by reverse transcriptase polymerase chain reaction.

text: OBJECTIVE To determine the frequencies of human rhinovirus (HRV), respiratory syncytial virus (RSV), and coronavirus (HCV) infection in children with acute otitis media (AOM). METHODS Middle ear fluids (MEF) collected by tympanocentesis and nasopharyngeal aspirates (NPA) at the time of the AOM diagnosis were examined by reverse transcriptase polymerase chain reaction for HRV, RSV, and HCV RNA. PATIENTS Ninety-two children aged 3 months to 7 years during a 1-year period. RESULTS Virus RNA was detected in a total of 69 children (75%) and in 44 MEF samples (48%) and 57 NPA samples (62%) at the time of AOM diagnosis. HRV RNA was detected in both MEF and NPA in 18 (20%), in MEF alone in 4 (4%), and in NPA alone in 10 (11%). RSV was detected in both MEF and NPA in 12 (13%), in MEF alone in 5 (5%), and in NPA alone in 9 (10%). HCV RNA was detected in both MEF and NPA in 5 (5%), in MEF alone in 2 (2%), and in NPA alone in 9 (10%). Dual viral infections were detected in 5% of children. HRV and RSV were detected simultaneously in 2 MEF samples and in 2 NPA samples; RSV and HCV were detected in 1 NPA sample. Bacterial pathogens were detected in 56 (62%) MEF from 91 children. Viral RNA was detected in 20 (57%) MEF of 35 bacteria-negative and in 25 (45%) of 56 bacteria-positive MEF samples. No important differences in the risk of treatment failure, relapse, or occurrence of late secretory otitis media were noted between children with virus-positive and virus-negative MEF aspirates. CONCLUSION These findings highlight the importance of common respiratory viruses, particularly HRV and RSV, in predisposing to and causing AOM in young children.


Sporadic contact with unfamiliar source makes epidemic unlikely.


title: Severe acute respiratory syndrome caused by the influenza A (H1N1) virus.

text: In view of the pandemic caused by a new virus, influenza A (H1N1), we report the case of a 56-year-old patient without relevant risk factors and with severe acute respiratory syndrome resulting from infection with this virus. We present the results of laboratory tests and the imaging findings (chest X-ray and CT scans). The evolution was favorable, and the patient was discharged after 14 days.


title: Evaluation and application of an indirect ELISA for the detection of antibodies to bovine respiratory syncytial virus in milk, bulk milk, and serum.

text: An indirect enzyme-linked immunosorbent assay (ELISA) was developed at the National Veterinary Institute (NVI), Uppsala, to detect antibodies to bovine respiratory syncytial virus (BRSV) in serum and milk. For the evaluation of the NVI ELISA, field sera collected from cattle in England and Sweden were tested in parallel with an ELISA in use at the Central Veterinary Laboratory (CVL), Weybridge. The tests showed 96% agreement. The sensitivity and specificity of the NVI ELISA relative to the CVL ELISA were 94% and 100%, respectively. There was evidence that the difference in sensitivity between the 2 tests was due to the detection of both IgG and IgM class antibodies by the CVL ELISA, whereas the NVI ELISA was designed specifically to detect IgG1. Milk and serum samples from individual cows were tested by the NVI ELISA for presence of antibodies to BRSV. There was a good correlation between the ability to detect antibodies in serum and the ability to detect them in milk, although the antibody titer was generally lower in milk than in serum. Bulk milk samples were collected from farms with severe clinical symptoms of respiratory distress and from farms with no history of respiratory disease. There was a clear distinction between antibody levels in diseased and healthy herds. The NVI ELISA is a rapid and reliable test for detecting antibodies to BRSV in milk, bulk milk, and serum samples.


title: Observations on the quasispecies composition of three animal pathogenic RNA viruses.

text: The quasispecies nature of three animal pathogenic RNA viruses of field origin was examined by testing variants of classical swine fever virus (CSFV) originating from geographically different areas, feline coronavirus (FCoV) detected from the same animal by successive sampling, and rabbit haemorrhagic disease virus (RHDV) originating from successive outbreaks in the same geographic area. Clinical samples were investigated using reverse transcriptase polymerase chain reaction (RT-PCR) and ensuing single strand conformational polymorphism (SSCP) assay. By the combination of these methods even subtle differences could be detected among the amplified fragments of the same virus species of different origin. FCoV proved to comprise the most and CSFV the less heterogeneous virus quasispecies. The results show that the combination of RT-PCR and SSCP provides novel and highly sensitive means for the characterisation of RNA viruses, with special regard to genome composition, evolution, features of pathogenicity and molecular epizootiology.


title: Immunological responses against SARS-coronavirus infection in humans.

text: Since the outbreak of a SARS epidemic last year, significant advances have been made on our understanding of the mechanisms of interaction between the SARS coronavirus (CoV) and the immune system. Strong humoral responses have been found in most patients following SARS-CoV infection, with high titers of neutralizing Abs present in their convalescent sera. The nucleocapsid (N) and spike (S) proteins of SARS-CoV appear to be the dominant antigens recognized by serum Abs. CD4+ T cell responses against the N protein have been observed in SARS patients and an HLA-A2-restricted cytotoxic T lymphocyte epitope in the S protein has been identified. It is likely that the immune responses induced by SARS-CoV infection could also cause pathological damage to the host, especially in the case of proinflammatory cytokines. There is also evidence suggesting that SARS-CoV might be able to directly invade cells of the immune system. Our understanding on the interaction between SARS-CoV, the immune system and local tissues is essential to future diagnosis, control and treatment of this very contagious disease.


Update on Middle East respiratory syndrome coronavirus (MERS-CoV).


title: Hyperbaric oxygen as a treatment for COVID-19 infection?

text: Recently the internet has been abuzz with new ideas to treat COVID-19, including hyperbaric oxygen (HBO2) therapy, undoubtedly driven by the fact that until recently there have been few therapeutic options for this highly contagious and often lethal infection. . . . Refractory hypoxemia is certainly treatable with hyperbaric oxygen due to the obvious effect of increasing inspired oxygen partial pressure (PO2), the major reason for using HBO2 for its established indications. However, the length of time during which patients can safely be administered HBO2 inside a chamber is limited, due to practical issues of confinement and isolation from other necessary medical interventions, but also because of oxygen toxicity.


title: Essential role for collectrin in renal amino acid transport.

text: Angiotensin -converting enzyme 2 (ACE2) is a regulator of the renin angiotensin system involved in acute lung failure, cardiovascular functions and severe acute respiratory syndrome (SARS) infections in mammals. A gene encoding a homologue to ACE2, termed collectrin (Tmem27), has been identified in immediate proximity to the ace2 locus. The in vivo function of collectrin was unclear. Here we report that targeted disruption of collectrin in mice results in a severe defect in renal amino acid uptake owing to downregulation of apical amino acid transporters in the kidney. Collectrin associates with multiple apical transporters and defines a novel group of renal amino acid transporters. Expression of collectrin in Xenopus oocytes and Madin-Darby canine kidney (MDCK) cells enhances amino acid transport by the transporter B(0)AT1. These data identify collectrin as a key regulator of renal amino acid uptake.


title: Visualization of double-stranded RNA in cells supporting hepatitis C virus RNA replication.

text: The mechanisms involved in hepatitis C virus (HCV) RNA replication are unknown, and this aspect of the virus life cycle is not understood. It is thought that virus-encoded nonstructural proteins and RNA genomes interact on rearranged endoplasmic reticulum (ER) membranes to form replication complexes, which are believed to be sites of RNA synthesis. We report that, through the use of an antibody specific for double-stranded RNA (dsRNA), dsRNA is readily detectable in Huh-7 cells that contain replicating HCV JFH-1 genomes but is absent in control cells. Therefore, as that of other RNA virus genomes, the replication of the HCV genome may involve the generation of a dsRNA replicative intermediate. In Huh-7 cells supporting HCV RNA replication, dsRNA was observed as discrete foci, associated with virus-encoded NS5A and core proteins and identical in morphology and distribution to structures containing HCV RNA visualized by fluorescence-based hybridization methods. Three-dimensional reconstruction of deconvolved z-stack images of virus-infected cells provided detailed insight into the relationship among dsRNA foci, NS5A, the ER, and lipid droplets (LDs). This analysis revealed that dsRNA foci were located on the surface of the ER and often surrounded, partially or wholly, by a network of ER-bound NS5A protein. Additionally, virus-induced dsRNA foci were juxtaposed to LDs, attached to the ER. Thus, we report the visualization of HCV-induced dsRNA foci, the likely sites of virus RNA replication, and propose that HCV genome synthesis occurs at LD-associated sites attached to the ER in virus-infected cells.


title: The role of biochemical markers as early indicators of cardiac damage and prognostic parameters of perinatal asphyxia.

text: BACKGROUND/AIM In recent years, the focus of interest of the scientific community is the application of heart markers as early indicators and prognostic parameters of perinatal asphyxia (PA). The aim of this study was to evaluate the significance of clinical application of heart markers in term newborns with perinatal asphyxia. METHODS During a 3-year period we analyzed 91 full-term newborns (55 with and 36 without perinatal asphyxia). In all the subjects within the first 24-48 h after birth, we simultaneously determined serum concentrations of cardiac troponin I, brain natriuretic peptide, MB fraction of creatine kinase (CK-MB) and C-reactive protein. RESULTS In the group of full-term neonates with PA significantly higher levels of cardiac tropon-inI (p = 0.000), CK-MB fraction (p = 0.000), brain natriuretic peptide (p = 0.003) and C-reactive protein (p = 0.017) were found, compared to the group of healthy full-term newborns. In merged group (n = 91) cardiac troponin I level correlated with the fifth minute Apgar score (r = -0.637, p = 0.000) and the serum lactate concentration in the first 12h after birth (r = 0.529, p = 0.000). Early increase in cardiac troponin I > 0.135 microg/L predicted the risk of death with the sensitivity of 84.6% and specificity of 85.9%, while the increase in CK-MB fraction, brain natriuretic peptide and C-reactive protein did not have a predictive value with respect to a mortality outcome. CONCLUSION Among the tested cardiac markers, cardiac troponin I is the most sensitive and the only reliable early predictor of mortality in full-term neonates with perinatal asphyxia.


title: Risk perception and impact of Severe Acute Respiratory Syndrome (SARS) on work and personal lives of healthcare workers in Singapore: what can we learn?

text: INTRODUCTION Healthcare workers (HCWs) were at the frontline during the battle against Severe Acute Respiratory Syndrome (SARS). Understanding their fears and anxieties may hold lessons for handling future outbreaks, including acts of bioterrorism. METHOD We measured risk perception and impact on personal and work life of 15,025 HCWs from 9 major healthcare institutions during the SARS epidemic in Singapore using a self-administered questionnaire and Impact of Events Scale and analyzed the results with bivariate and multivariate statistics. RESULTS From 10,511 valid questionnaires (70% response), we found that although the majority (76%) perceived a great personal risk of falling ill with SARS, they (69.5%) also accepted the risk as part of their job. Clinical staff (doctors and nurses), staff in daily contact with SARS patients, and staff from SARS-affected institutions expressed significantly higher levels of anxiety. More than half reported increased work stress (56%) and work load (53%). Many experienced social stigmatization (49%) and ostracism by family members (31%), but most (77%) felt appreciated by society. Most felt that the personal protective measures implemented were effective (96%) and that the institutional policies and protocols were clear (93%) and timely (90%). CONCLUSION During epidemics, healthcare institutions have a duty to protect HCWs and help them cope with their personal fears and the very stressful work situation. Singapore's experience shows that simple protective measures based on sound epidemiological principles, when implemented in a timely manner, go a long way to reassure HCWs.


title: Transmission of sialodacryoadenitis virus (SDAV) from infected rats to rats and mice through handling, close contact, and soiled bedding.

text: Thirty mice and six rats were exposed through handling, soiled bedding, or close contact to rats previously inoculated with sialodacryoadenitis virus (SDAV). All exposed rats developed coronaviral antibody without clinical signs or lesions of SDAV infection. Exposed mice had no lesions or clinical signs of coronavirus infection. Mice exposed by handling or by soiled bedding did not develop coronavirus antibody. Two of 10 mice exposed to SDAV-inoculated rats by close contact were coronavirus seropositive when tested 3 weeks postexposure. SDAV-inoculated rats and mice developed coronavirus lesions and antibody. These results suggest that rat-to-rat transmission of SDAV is likely via fomites or handling; however, rat-to-mouse transmission is unlikely when animals are housed and husbanded using modern techniques. Results also suggest that coronavirus antibody in mice is due to exposure to mouse coronavirus and not to rat coronaviruses.


title: A cis-acting function for the coronavirus leader in defective interfering RNA replication.

text: To test the hypothesis that the 65-nucleotide (nt) leader on subgenomic mRNAs suffices as a 5'-terminal cis-acting signal for RNA replication, a corollary to the notion that coronavirus mRNAs behave as replicons, synthetic RNA transcripts of a cloned, reporter-containing N mRNA (mRNA 7) of the bovine coronavirus with a precise 5' terminus and a 3' poly(A) of 68 nt were tested for replication after being transfected into helper virus-infected cells. No replication was observed, but synthetic transcripts of a cloned reporter-containing defective interfering (DI) RNA differing from the N mRNA construct by 433 nt of continuous 5'-proximal genomic sequence between the leader and the N open reading frame did replicate and become packaged, indicating the insufficiency of the leader alone as a 5' signal for replication of transfected RNA molecules. The leader was shown to be a necessary part of the cis-acting signal for DI RNA replication, however, since removal of terminal bases that destroyed a predicted intraleader stem-loop also destroyed replicating ability. Surprisingly, when the same stem-loop was disrupted by base substitutions, replication appeared only minimally impaired and the leader was found to have rapidly reverted to wild type during DI RNA replication, a phenomenon reminiscent of high-frequency leader switching in the mouse hepatitis coronavirus. These results suggest that once a minimal structural requirement for leader is fulfilled for initiation of DI RNA replication, the wild-type leader is strongly preferred for subsequent replication. They also demonstrate that, in contrast to reported natural mouse hepatitis coronavirus DI RNAs, the DI RNA of the bovine coronavirus does not require sequence elements originating from discontinuous downstream regions within the polymerase gene for replication or for packaging.


title: A novel coronavirus associated with severe acute respiratory syndrome.

text: BACKGROUND A worldwide outbreak of severe acute respiratory syndrome (SARS) has been associated with exposures originating from a single ill health care worker from Guangdong Province, China. We conducted studies to identify the etiologic agent of this outbreak. METHODS We received clinical specimens from patients in seven countries and tested them, using virus-isolation techniques, electron-microscopical and histologic studies, and molecular and serologic assays, in an attempt to identify a wide range of potential pathogens. RESULTS None of the previously described respiratory pathogens were consistently identified. However, a novel coronavirus was isolated from patients who met the case definition of SARS. Cytopathological features were noted in Vero E6 cells inoculated with a throat-swab specimen. Electron-microscopical examination revealed ultrastructural features characteristic of coronaviruses. Immunohistochemical and immunofluorescence staining revealed reactivity with group I coronavirus polyclonal antibodies. Consensus coronavirus primers designed to amplify a fragment of the polymerase gene by reverse transcription-polymerase chain reaction (RT-PCR) were used to obtain a sequence that clearly identified the isolate as a unique coronavirus only distantly related to previously sequenced coronaviruses. With specific diagnostic RT-PCR primers we identified several identical nucleotide sequences in 12 patients from several locations, a finding consistent with a point-source outbreak. Indirect fluorescence antibody tests and enzyme-linked immunosorbent assays made with the new isolate have been used to demonstrate a virus-specific serologic response. This virus may never before have circulated in the U.S. population. CONCLUSIONS A novel coronavirus is associated with this outbreak, and the evidence indicates that this virus has an etiologic role in SARS. Because of the death of Dr. Carlo Urbani, we propose that our first isolate be named the Urbani strain of SARS-associated coronavirus.


title: Detection of bovine coronavirus and type A rotavirus in neonatal calf diarrhea and winter dysentery of cattle in Quebec: evaluation of three diagnostic methods.

text: The use of direct electron microscopy, enzyme-linked immunosorbent assay, and protein A-gold immunoelectron microscopy for the identification of bovine coronavirus and type A rotavirus were examined. Two hundred and forty-nine samples from diarrheic calves and winter dysenteric cattle from seven geographic areas in Quebec were examined for the presence of viruses by direct electron microscopy of negatively stained preparations. In addition, all the samples were analyzed by enzyme-linked immunosorbent assay, and a random selection of 47 samples were also analyzed by protein A-gold immunoelectron microscopy. Thirty-nine percent of samples examined by direct electron microscopy contained viral particles; bovine coronavirus and type A rotavirus were the most common viruses involved. Overall agreement between any two of the methods used compared favorably with results obtained by others using similar methods. The presence of coronavirus and rotavirus in fecal samples obtained from neonatal calves and the presence of coronavirus in samples from winter dysenteric adult cattle suggested their etiological roles in the respective diseases. Furthermore, results from protein A-gold immunoelectron microscopy of coronavirus-like particles implied that a different coronavirus or some other viruses might be involved in these diseases. Finally, the efficiency of direct electron microscopy, enzyme-linked immunosorbent assay and protein A-gold immunoelectron microscopy as diagnostic tools is discussed.


title: [Psychological and physical presentations of severe acute respiratory syndrome].

text: OBJECTIVE To study the relation between psychological symptoms and physical parameters in patients with severe acute respiratory syndrome (SARS) at different stages of the disease. METHODS Physical parameters such as SCL-90, chest CT, T lymphocytes and subset and SaO2 were studied in 29 SARS patients at three different stages (initial stage, serious stage, rehabilitating stage). RESULTS Patients with SARS showing severe psychological symptoms constituted about 41% (group I) and patients with SARS showing mild psychological symptoms (group II) constituted about 59%. There were no change of the psychological symptoms at the three stages in group I and II (P > 0.05). Statistical differences were found between physical parameters of group I and II in rehabilitating stage. CONCLUSIONS Severe psychological symptoms such as obsession, depression and anxiety were found in a minority of patients with SARS and the psychological symptoms were not related with the change of physical parameters. The majority of the patients with SARS did not have severe psychological symptoms. Whether or not psychological symptoms were related to their personality needs further study.


title: Non-superiority of Kakkonto, a Japanese herbal medicine, to a representative multiple cold medicine with respect to anti-aggravation effects on the common cold: a randomized controlled trial.

text: OBJECTIVE Kakkonto, a Japanese herbal medicine, is frequently used to treat the common cold not only with a physician's prescription, but also in self-medication situations. This study aimed to examine whether Kakkonto prevents the aggravation of cold symptoms if taken at an early stage of illness compared with a well-selected Western-style multiple cold medicine. METHODS This study was a multicenter, active drug-controlled, randomized trial. Adults 18 to 65 years of age who felt a touch of cold symptoms and visited 15 outpatient healthcare facilities within 48 hours of symptoms onset were enrolled. The participants were randomly assigned to two groups: one treated with Kakkonto (Kakkonto Extract-A, 6 g/day) (n=209) and one treated with a Western-style multiple cold medicine (Pabron Gold-A, 3.6 g/day) (n=198) for at most four days. The primary outcome of this study was the aggravation of cold, nasal, throat or bronchial symptoms, scored as moderate or severe and lasting for at least two days within five days after entry into the study. RESULTS Among the 410 enrollees, 340 (168 in the Kakkonto group and 172 in the Pabron group) were included in the analyses. The proportion of participants whose colds were aggravated was 22.6% in the Kakkonto group and 25.0% in the Pabron group (p=0.66). The overall severity of the cold symptoms was not significantly different between the groups. No harmful adverse events occurred in either group. CONCLUSION Kakkonto did not significantly prevent the progression of cold symptoms, even when prescribed at an early stage of the disease.


title: Classification of IBV S1 genotypes by direct reverse transcriptase-polymerase chain reaction (RT-PCR) and relationship between serotypes and genotypes of strains isolated between 1998 and 2008 in Japan.

text: In this study, we attempted to establish a simple detection method for classification of IBV S1 genotypes by direct reverse transcriptase-polymerase chain reaction (RT-PCR). Then, to evaluate the usefulness of the S1 genotype-specific RT-PCR, we examined the relationship between S1 genotypes and serotypes of IBV in Japan. Sequencing of the S1 genes of IBV and phylogenetic tree analysis were conducted. On the basis of the sequencing data of the S1 genotype samples, we determined primer sets specific for each genotype. Five vaccine strains in Japan as reference strains and 46 field isolates were classified into different genetic clusters by phylogenetic tree analysis (JP-1, JP-II, JP-III, Mass and 4/91) and were matched to the results of S1 genotype-specific RT-PCR. A cross virus-neutralizing test showed that the five vaccine strains in Japan exhibited different serotypes from each other. The concordance rate of the 46 field isolates between the S1 genotypes and serotypes was 65.2%. The present study indicates that genotype-specific RT-PCR could be a convenient and useful tool for determining IBV serotypes and could contribute to the control of IBV outbreaks in Japan.


title: [The use of the fluorescent probe method for the diagnosis of acute coronavirus intestinal infections in people].

text: The scientific investigations conducted first allow using a method of fluorescent probes for the diagnosis of human coronaviral intestinal infection. The method is characterised by sensitivity, specificity, reproductivity and proximity. It is comparable, according to these parameters, with the methods of the third generation and may be recommended for putting into practical work of virological laboratories.


title: Molecular epidemiology of chicken anemia virus (CAV) in southeastern Chinese live birds markets.

text: Between January 2004 and December 2005, cloacal swabs from essentially healthy chickens and silky chickens from live birds markets in Guangdong and Hunan provinces in southeastern China were screened for chicken anemia virus (CAV) by polymerase chain reaction. Phylogenetic analysis of the major structural protein VP1 sequences showed no clear genotype cluster and no correlation with the geographic origin of CAV strains. Virus evolution at the amino acid level was very slow, which corresponds to a strong negative selection of the VP1 gene in China and worldwide. A high proportion (87%) of birds was CAV positive, suggesting that many farms in the region were infected. Further investigations are necessary to evaluate the economic losses caused by CAV and the cost-benefit of vaccination.