title: Self-directed practice schedule enhances learning of suturing skills.

text: BACKGROUND Most preoperative surgical training programs experience challenges with the availability of expert surgeons to teach trainees. Some research suggests that trainees may benefit from being allowed to actively shape their learning environments, which could alleviate some of the time and resource pressures in surgical training. The purpose of this study was to investigate the effects of self-directed or prescribed practice schedules (random or blocked) on learning suturing skills. METHODS Participants watched an instructional video for simple interrupted, vertical mattress and horizontal mattress suturing then completed a pretest to assess baseline skills. Participants were assigned to 1 of 4 practice groups: self-directed practice schedule, prescribed blocked practice schedule, prescribed random practice schedule or matched to the self-directed group (control). Practice of the skill was followed by a delayed (1 h) posttest. Improvement from pretest to posttest was determined based on differences in performance time and expert-based assessments. RESULTS Analyses revealed a significant effect of group for difference in performance time of the simple interrupted suture. Random practice did not show the expected advantage for skill learning, but there was an advantage of self-directed practice. CONCLUSION Self-directed practice schedules may be desirable for optimal learning of simple technical skills, even when expert instruction is available. Instructors must also take into account the interaction between task difficulty and conditions of practice to develop ideal training environments.


title: The impact of viral respiratory tract infections on long-term morbidity and mortality following lung transplantation: a retrospective cohort study using a multiplex PCR panel.

text: BACKGROUND The major factor affecting morbidity and mortality after lung transplantation (LTX) is bronchiolitis obliterans syndrome. Earlier studies have suggested a connection between the presence of viral agents and morbidity in this patient group, but data are somewhat conflicting. The objective of this study was to investigate the development of bronchiolitis obliterans syndrome and graft loss after LTX in relation to the presence of respiratory viruses during the first year after LTX. METHOD The study is a retrospective cohort study of 39 LTX recipients 11Y13 years after surgery. Patients were operated between January 1, 1998 and December 31, 2000 at Sahlgrenska University Hospital. The presence of virus in bronchoalveolar lavage (BAL) fluids from patients during the first year after surgery was analyzed retrospectively using a multiplex polymerase chain reaction test capable of detecting 15 respiratory agents. The time to BOS or graft loss was analyzed in relation to the positive findings in BAL during the first year after LTX. RESULTS Patients with one or more viruses detected in BAL during the first year after transplantation demonstrated a significantly faster development of BOS (P=0.005) compared with patients with no virus detected. No significant difference in graft survival was found. CONCLUSION Our results suggest that the long-term prognosis after LTX may be negatively affected by viral respiratory tract infections during the first year after LTX.


title: Clot Burden Score on Baseline Computerized Tomographic Angiography and Intra-Arterial Treatment Effect in Acute Ischemic Stroke.

text: BACKGROUND AND PURPOSE A high clot burden score (CBS) is associated with favorable outcome after intravenous treatment for acute ischemic stroke. The added benefit of intra-arterial treatment might be less in these patients. The aim of this exploratory post hoc analysis was to assess the relation of CBS with neurological improvement and endovascular treatment effect. METHODS For 499 of 500 patients in the MR CLEAN study (Multicenter Randomized Clinical Trial of Endovascular Treatment for Acute Ischemic Stroke in the Netherlands), the CBS was determined. Ordinal logistic regression models with and without main baseline prognostic variables were used to assess the association between CBS (continuous or dichotomized at CBS of 6) and a shift toward better outcome on the modified Rankin Scale. The model without main baseline prognostic variables only included treatment allocation and CBS. Models with and without a multiplicative interaction term of CBS and treatment were compared using the χ2 test to assess treatment effect modification by CBS. RESULTS Higher CBS was associated with a shift toward better outcome on the modified Rankin Scale; adjusted common odds ratio per point CBS was 1.12 (95% confidence interval, 1.04-1.20]. Dichotomized CBS had an adjusted common odds ratio of 1.67 (95% confidence interval, 1.12-2.51). Both effect estimates were slightly attenuated by adding baseline prognostic variables. The addition of the interaction terms did not significantly improve the fit of the models. There was a small and insignificant increase of intra-arterial treatment efficacy in the high CBS group. CONCLUSIONS A higher CBS is associated with improved outcome and may be used as a prognostic marker. We found no evidence that CBS modifies the effect of intra-arterial treatment. CLINICAL TRIAL REGISTRATION URL: http://www.trialregister.nl. Unique identifier: NTR1804. URL: http://www.controlled-trials.com. Unique identifier: ISRCTN10888758.


title: The sequence of the full spike S1 glycoprotein of infectious bronchitis virus circulating in Egypt reveals evidence of intra-genotypic recombination.

text: Infectious bronchitis virus (IBV) continues to circulate worldwide, with a significant impact on the poultry industry and affecting both vaccinated and unvaccinated flocks. Several studies have focused on the hypervariable regions (HVRs) of the spike gene (S1); however, genetic and bioinformatics studies of the whole S1 gene are limited. In this study, the whole S1 gene of five Egyptian IBVs was genetically analyzed. Phylogenetic analysis revealed that the Egyptian IBVs are clustered within two distinct groups: the classic group resembling the GI-1 genotype (vaccine strains) and the variant group (field strains) of the GI-23 genotype. The variant genotype was divided into two distinct subgroups (Egy/var I and Egy/var II) resembling the Israeli variants IS/1494 and IS885 strain, respectively. Significant amino acid sequence differences between the two subgroups, especially in the epitope sites, were identified. A deletion at position 63 and an I69A/S substitution mutation associated with virus tropism were detected in the receptor-binding sites. The deduced amino acid sequence of HVRs of the variant subgroups indicated different genetic features in comparison to the classic vaccine group (H120 lineage). The Egyptian variant IBVs also contained additional N-glycosylation sites compared to the classical viruses. Recombination analysis gave evidence for distinct patterns of origin by recombination throughout the S1 gene, suggesting that the recent virus IBV-EG/1586CV-2015 emerged as a recombinant of two viruses from the variant groups Egy/var I and Egy/var II, providing another example of intra-genotypic recombination among IBVs and the first example of recombination within the GI-23 genotype. Our data suggest that both mutation and recombination may be contributing to the emergence of IBV variants. Moreover, we found that the commercially used vaccines are genotypically distant from the circulating field strains. Hence, continuous follow-up of the current vaccine strategy is highly recommended for better control and prevention of infectious bronchitis virus in the poultry sector in Egypt.


title: Efficacy of an inactivated canine coronavirus vaccine in pups.

text: The efficacy of an inactivated CCoV vaccine (Duramune PC) was evaluated in four pups. Two dogs were maintained non-vaccinated. Ten days after the booster shot all the pups were challenged with a field CCoV strain administered by oro-nasal route. The vaccinated pups did not display clinical signs and shed the challenge-virus for 11.25 days, evaluated by virus isolation, and 13.5 days, evaluated by PCR assay. The two non vaccinated pups displayed mild diarrhoea at day post-challenge 4 and shed the challenge-virus for 14 and 15 days respectively, by virus isolation, and for 22 and 24 days respectively, by PCR assay.


COVID-19: another infectious disease emerging at the animal-human interface.


title: Frontiers in nephrology: heterologous immunity, T cell cross-reactivity, and alloreactivity.

text: Established memory T cell responses to a previously encountered pathogen can have a major impact on the course and outcome of a subsequent infection with an unrelated pathogen. This phenomenon, known as heterologous immunity, is dependent on the sequence of infections and can be either beneficial or detrimental to the host. Examples of heterologous immunity between unrelated viruses and alloantigens are mounting, and the role of cross-reactive T cells both in the pathogenesis of infections and in transplant rejection is now being explored. Memory T cells seem to be part of a continually evolving interactive network in which with each new infection, an alteration in the frequencies, distributions, and activities of memory cells is generated in response to previous infections and alloantigens.


title: Severe acute respiratory syndrome--retrospect and lessons of 2004 outbreak in China.

text: OBJECTIVE To summarize lessons learned from an outbreak of severe acute respiratory syndrome (SARS) in China during the spring of 2004. METHODS Data of SARS cases were officially reported by Beijing Municipal Center for Disease Control and Prevention (BCDC) and Anhui Provincial Center for Disease Control and Prevention (APCDC) and results of epidemiological investigations were collected and analyzed. RESULTS Three generations of 11 cases of SARS were identified during the outbreak. Initial two cases were most likely to be infected in Diarrhea Virus Laboratory of National Institute of Virology, China Centers for Disease Control and Prevention and main mode of transmission was direct contact with SARS patients. Delay in detecting initial case resulted in spread of the illness at hospitals and communities with two generations of secondary cases. CONCLUSIONS SARS outbreak in 2004 has yielded following lessons for public health globally. (1) Lab bio-safety programs should be made and should be strictly abided by. Studies in highly pathogenic viruses such as SARS coronavirus should be utmost cautious. (2) Management systems of occupational exposure to virus and disease surveillance need to be strengthened to take all risk factors into account so as to detect potential patients with infectious disease as early as possible.


title: Comparison of Absorbable Versus Nonabsorbable Tackers in Terms of Long-term Outcomes, Chronic Pain, and Quality of Life After Laparoscopic Incisional Hernia Repair: A Randomized Study.

text: BACKGROUND Laparoscopic incisional and ventral hernia repair (LIVHR) has been associated with a high incidence acute and chronic pain due to use of nonabsorbable tackers. Several absorbable tackers have been introduced to overcome these complications. This randomized study was done to compare 2 techniques of mesh fixation, that is, nonabsorbable versus absorbable tackers for LIVHR. MATERIALS AND METHODS Ninety patients admitted for LIVHR repair (defect size <15 cm) were randomized into 2 groups: nonabsorbable tacker fixation (NAT group, 45 patients) and absorbable tacker fixation (AT group, 45 patients). Intraoperative variables and postoperative outcomes were recorded and analyzed. RESULTS Patients in both the groups were comparable in terms of demographic profile and hernia characteristics. Mesh fixation time and operation time were also comparable. There was no significant difference in the incidence of immediate postoperative and chronic pain over a mean follow-up of 8.8 months. However, cost of the procedure was significantly higher in AT group (P<0.01) and NAT fixation was more cost effective as compared with AT. Postoperative quality of life outcomes and patient satisfaction scores were also comparable. CONCLUSIONS NAT is a cost-effective method of mesh fixation in patients undergoing LIVHR with comparable early and late postoperative outcomes in terms of pain, quality of life, and patient satisfaction scores.


docid: qglqxsfr

title: Function of a 5'-end genomic RNA mutation that evolves during persistent mouse hepatitis virus infection in vitro.

text: Persistently infected cultures of DBT cells were established with mouse hepatitis virus strain A59 (MHV-A59), and the evolution of the MHV leader RNA and 5' end of the genome was studied through 119 days postinfection. Sequence analysis of independent clones demonstrated an overall mutation frequency approaching 1.2 x 10(-3) to 6.7 x 10(-3). The rate of fixation of mutations was about 1.2 x 10(-5) to 7.6 x 10(-5) per nucleotide (nt) per day. In contrast to finding in bovine coronavirus, the MHV leader RNA sequences were extremely stable and did not evolve significantly during persistent infection. Rather, a 5' untranslated region (UTR) A-to-G mutation at nt 77 in the genomic RNA emerged by day 56 and accumulated until 50 to 80% of the genome-length molecules retained the mutation by 119 days postinfection. Although other 5'-end mutations were noted, only the nt 77 mutation was significantly associated with viral persistence in vitro. Mutations were also found in the 5' end of the p28 coding region, but no specific alterations accumulated in genome-length molecules through 119 days postinfection. The 5' UTR nt 77 mutation resulted in an 18-amino-acid open reading frame (ORF) upstream of the ORF 1a AUG start site. By in vitro translation assays, the small ORF was not translated into detectable product but the mutation significantly enhanced translation of the downstream p28 ORF about 2.5-fold. Variant viruses, containing either the nt 77 A-to-G mutation (V16-ATG+) or wild-type sequences at this locus (V1-ATG-), were isolated at 119 days postinfection. The variant viruses replicated more efficiently than wild-type virus and were extremely cytolytic in DBT cells, suggesting that the A-to-G mutation did not encode a nonlytic or attenuated phenotype. Consistent with the in vitro translation results, a significant increase (approximately 3.5-fold) in p28 expression was also observed with the mutant virus (V16-ATG+) in DBT cells compared with that in wild-type controls. These data indicate that MHV persistence was significantly associated with mutation and evolution in the 5'-end UTR which enhanced the translation of the ORF 1a and potentially ORF 1b polyproteins which function in virus transcription and replication.


Some thoughts on COVID-19.


Cost-effectiveness of influenza vaccination for elderly people living in the community.


title: Update: severe acute respiratory syndrome--United States, June 11, 2003.

text: CDC continues to work with state and local health departments, the World Health Organization (WHO), and other partners to investigate cases of severe acute respiratory syndrome (SARS). This report updates SARS cases reported worldwide and in the United States, and describes the eighth probable U.S. SARS case with laboratory evidence of SARS-associated coronavirus (SARS-CoV) infection.


title: Recombinant modified vaccinia virus Ankara expressing the spike glycoprotein of severe acute respiratory syndrome coronavirus induces protective neutralizing antibodies primarily targeting the receptor binding region.

text: Immunization with a killed or inactivated viral vaccine provides significant protection in animals against challenge with certain corresponding pathogenic coronaviruses (CoVs). However, the promise of this approach in humans is hampered by serious concerns over the risk of leaking live severe acute respiratory syndrome (SARS) viruses. In this study, we generated a SARS vaccine candidate by using the live-attenuated modified vaccinia virus Ankara (MVA) as a vector. The full-length SARS-CoV envelope Spike (S) glycoprotein gene was introduced into the deletion III region of the MVA genome. The newly generated recombinant MVA, ADS-MVA, is replication incompetent in mammalian cells and highly immunogenic in terms of inducing potent neutralizing antibodies in mice, rabbits, and monkeys. After two intramuscular vaccinations with ADS-MVA alone, the 50% inhibitory concentration in serum was achieved with reciprocal sera dilutions of more than 1,000- to 10,000-fold in these animals. Using fragmented S genes as immunogens, we also mapped a neutralizing epitope in the region of N-terminal 400 to 600 amino acids of the S glycoprotein (S400-600), which overlaps with the angiotensin-converting enzyme 2 (ACE2) receptor-binding region (RBR; S318-510). Moreover, using a recombinant soluble RBR-Fc protein, we were able to absorb and remove the majority of the neutralizing antibodies despite observing that the full S protein tends to induce a broader spectrum of neutralizing activities in comparison with fragmented S proteins. Our data suggest that a major mechanism for neutralizing SARS-CoV likely occurs through blocking the interaction between virus and the cellular receptor ACE2. In addition, ADS-MVA induced potent immune responses which very likely protected Chinese rhesus monkeys from pathogenic SARS-CoV challenge.


title: Alignment-free comparison of protein sequences based on reduced amino acid alphabets.

text: Protein sequences are treated as stochastic processes on the basis of a reduced amino acid alphabet of 10 types of amino acids. The realization of a stochastic process is described by associated transition probability matrix that corresponds to the process uniquely. Then new distances between transition probability matrices are defined for sequences similarity analysis. Two separate datasets are prepared and tested to identify the validity of the method. The results demonstrate the new method is powerful and efficient.


title: Construction of a eukaryotic expression plasmid encoding partial S gene fragments of the SARS-CoV and its potential utility as a DNA vaccine.

text: The spike (S) protein, a main surface antigen of the SARS coronavirus (SARS-CoV), is considered to be one of the most important protective antigen candidates for targets for vaccine design against the virus. In this study, a secreted recombinant expression plasmid, pVAX-S1, encoding the partial S protein with a signal peptide, was constructed and used to immunize BALB/c mice through electroporation. It was demonstrated that the eukaryotic expression vector pVAX-S1 was successfully constructed by restriction enzyme and sequence analysis. The expressed protein could be detected specifically by Western blot analysis. The serum IgG level of the vaccine group mice was significantly higher than that of the corresponding control group at day 14 after vaccination (P < 0.05). The vaccine group demonstrated significantly higher S1 protein lymphocyte proliferation index (LPI) than the control groups (P < 0.05). Furthermore, in the experimental group, a decrease in the ratio of CD4(+) to CD8(+) T-lymphocytes and an increase level of IFN-gamma in serum were observed. However, interleukin-4 (IL-4) was not detectable in two groups. These results strongly demonstrated that the pVAX-S1 plasmid could induce humoral and cellular immune responses in mice, and may be a potential candidate for a DNA vaccine against the SARS coronavirus.


docid: udgjoxip

title: The emergence of porcine circovirus 2b genotype (PCV-2b) in swine in Canada.

text: Since late 2004, the swine industry in the province of Quebec has experienced a significant increase in death rate related to postweaning multisystemic wasting syndrome (PMWS). To explain this phenomenon, 2 hypotheses were formulated: 1) the presence of a 2nd pathogen could be exacerbating the porcine circovirus 2 (PCV-2) infection, or 2) a new and more virulent PCV-2 strain could be infecting swine. In 2005, 13 PMWS cases were submitted to the Quebec provincial diagnostic laboratory and PCV-2 was the only virus that could be found consistently by PCR in all 13 samples. The PCR detection results obtained for other viruses revealed the following: 61.5% were positive for porcine reproductive and respiratory syndrome virus, 30.8% for swine influenza virus, 15.4% for porcine parvovirus, 69.2% for swine torque teno virus (swTTV), 38.5% for swine hepatitis E virus (swHEV) and 84.6% for Mycoplasma hyorhinis; transmissible gastroenteritis virus and porcine respiratory coronavirus (TGEV/PRCV) was not detected. Sequences of the entire genome revealed that these PCV-2 strains belonged to a genotype (named PCV-2b) that has never been reported in Canada. Further sequence analyses on 83 other Canadian PCV-2 positive cases submitted to the provincial diagnostic laboratory during years 2005 and 2006 showed that 79.5% of the viral sequences obtained clustered in the PCV-2b genotype. The appearance of the PCV-2b genotype in Canada may explain the death rate increase related to PMWS, but this relationship has to be confirmed.


title: Viral etiology in infants hospitalized for acute bronchiolitis.

text: Acute bronchiolitis is predominantly a viral disease. Respiratory syncytial virus is the most common agent, but other newly identified viruses have also been considered as causes. The aim of the present study is to determine the respiratory viruses causing acute bronchiolitis in hospitalized infants. Infants younger than 2 years of age who were hospitalized for acute viral bronchiolitis in a children's hospital between November 2011 and May 2012 were evaluated for the presence of viruses as etiologic agents using a realtime polymerase chain reaction method.A total of 55 infants were included in this study. The mean age of the children was 6.98±5.53 months, and 63.6% were male. In the 55 children, 63 viruses were detected. A single viral pathogen was detected in 47 (85.5%) patients, and two viruses were co-detected in 8 (14.6%) patients. Respiratory syncytial virus was the most common virus identified, accounting for 25 (45.5%) cases, followed by rhinovirus (n=9, 16.4%), and human metapneumovirus (n = 8, 14.5%).Although respiratory syncytial virus remains the major viral pathogen in infants hospitalized for acute broncholitis, more than half of bronchiolitis cases are associated with other respiratory viruses.


title: The crystal structures of severe acute respiratory syndrome virus main protease and its complex with an inhibitor.

text: A newly identified severe acute respiratory syndrome coronavirus (SARS-CoV), is the etiological agent responsible for the outbreak of SARS. The SARS-CoV main protease, which is a 33.8-kDa protease (also called the 3C-like protease), plays a pivotal role in mediating viral replication and transcription functions through extensive proteolytic processing of two replicase polyproteins, pp1a (486 kDa) and pp1ab (790 kDa). Here, we report the crystal structures of the SARS-CoV main protease at different pH values and in complex with a specific inhibitor. The protease structure has a fold that can be described as an augmented serine-protease, but with a Cys-His at the active site. This series of crystal structures, which is the first, to our knowledge, of any protein from the SARS virus, reveal substantial pH-dependent conformational changes, and an unexpected mode of inhibitor binding, providing a structural basis for rational drug design.


title: Involvement of the platelet-activating factor receptor in host defense against Streptococcus pneumoniae during postinfluenza pneumonia.

text: Although influenza infection alone may lead to pneumonia, secondary bacterial infections are a much more common cause of pneumonia. Streptococcus pneumoniae is the most frequently isolated causative pathogen during postinfluenza pneumonia. Considering that S. pneumoniae utilizes the platelet-activating factor receptor (PAFR) to invade the respiratory epithelium and that the PAFR is upregulated during viral infection, we here used PAFR gene-deficient (PAFR-/-) mice to determine the role of this receptor during postinfluenza pneumococcal pneumonia. Viral clearance was similar in wild-type and PAFR-/- mice, and influenza virus was completely removed from the lungs at the time mice were inoculated with S. pneumoniae (day 14 after influenza infection). PAFR-/- mice displayed a significantly reduced bacterial outgrowth in their lungs, a diminished dissemination of the infection, and a prolonged survival. Pulmonary levels of IL-10 and KC were significantly lower in PAFR-/- mice, whereas IL-6 and TNF-alpha were only trendwise lower. These data indicate that the pneumococcus uses the PAFR leading to severe pneumonia in a host previously exposed to influenza A.


COVID-19: A global public health disaster.


title: Conservative management of pyothorax in an Amur tiger (Panthera tigris altaica).

text: Therapy for pyothorax, or pleural empyema, has not been described for large felids. This case describes the successful treatment of pyothorax in a captive, large felid. A 15-yr-old multiparous, female Amur tiger (Panthera tigris altaica) presented with nonspecific clinical signs caused by an insidious onset of pyothorax. Management of pyothorax cases in companion animals often involves thoracostomy tube placement with recurrent drainage of the pleural cavity, intensive supportive care, and monitoring. In this case, conservative management was elected because hospitalization was not a viable option. Thoracocentesis was performed to drain as much fluid from the chest cavity as possible, yielding more than 1.3 L. Corynebacterium sp. and unidentified anaerobic gram-positive cocci were cultured. Treatment included cefovecin subcutaneously, oral antibiotic therapy with clindamycin and marbofloxacin, meloxicam, and restricted exercise by minimizing access to the main exhibit. Significant improvement was noted clinically and radiographically 6 wk later, and no relapses were noted in the following weeks. An examination 11.5 mo later confirmed resolution.


title: Human viral gastroenteritis.

text: During the last 15 years, several different groups of fastidious viruses that are responsible for a large proportion of acute viral gastroenteritis cases have been discovered by the electron microscopic examination of stool specimens. This disease is one of the most prevalent and serious clinical syndromes seen around the world, especially in children. Rotaviruses, in the family Reoviridae, and fastidious fecal adenoviruses account for much of the viral gastroenteritis in infants and young children, whereas the small caliciviruses and unclassified astroviruses, and possibly enteric coronaviruses, are responsible for significantly fewer cases overall. In addition to electron microscopy, enzyme immunoassays and other rapid antigen detection systems have been developed to detect rotaviruses and fastidious fecal adenoviruses in the stool specimens of both nonhospitalized patients and those hospitalized for dehydration and electrolyte imbalance. Experimental rotavirus vaccines have also been developed, due to the prevalence and seriousness of rotavirus infection. The small, unclassified Norwalk virus and morphologically similar viruses are responsible for large and small outbreaks of acute gastroenteritis in older children, adolescents, and adults. Hospitalization of older patients infected with these viruses is usually not required, and their laboratory diagnoses have been limited primarily to research laboratories.


title: A case study of the evaluation of a public health intervention.

text: Large public health interventions to control infectious disease outbreaks are common, but rigorous evaluation to improve the quality and effectiveness of these is rarely undertaken. Following a large community-based clinic to prevent a hepatitis A outbreak, a multifaceted and multidisciplinary evaluation was conducted involving consumers, health professionals and industry partners. The results of this evaluation were used to produce practical operational guidelines for the planning and conduct of future interventions. These guidelines have been distributed to all public health units in New South Wales and may be included in the next edition of the NSW Health notifiable diseases manual. The evaluation approach can be applied to all public health interventions across NSW and Australia to assist in the development of operational guidelines, in order to increase the quality of public health action in outbreak prevention.


title: Persistent infection of human oligodendrocytic and neuroglial cell lines by human coronavirus 229E.

text: Human coronaviruses (HuCV) cause common colds. Previous reports suggest that these infectious agents may be neurotropic in humans, as they are for some mammals. With the long-term aim of providing experimental evidence for the neurotropism of HuCV and the establishment of persistent infections in the nervous system, we have evaluated the susceptibility of various human neural cell lines to acute and persistent infection by HuCV-229E. Viral antigen, infectious virus progeny and viral RNA were monitored during both acute and persistent infections. The astrocytoma cell lines U-87 MG, U-373 MG, and GL-15, as well as neuroblastoma SK-N-SH, neuroglioma H4, and oligodendrocytic MO3.13 cell lines, were all susceptible to an acute infection by HuCV-229E. The CHME-5 immortalized fetal microglial cell line was not susceptible to infection by this virus. The MO3.13 and H4 cell lines also sustained a persistent viral infection, as monitored by detection of viral antigen and infectious virus progeny. Sequencing of the S1 gene from viral RNA after approximately 130 days of infection showed two point mutations, suggesting amino acid changes during persistent infection of MO3.13 cells but none for H4 cells. Thus, persistent in vitro infection did not generate important changes in the S1 portion of the viral spike protein, which was shown for murine coronaviruses to bear hypervariable domains and to interact with cellular receptor. These results are consistent with the potential persistence of HuCV-229E in cells of the human nervous system, such as oligodendrocytes and possibly neurons, and the virus's apparent genomic stability.


docid: hanhrqup

title: Pattern recognition methods for protein functional site prediction.

text: Protein functional site prediction is closely related to drug design, hence to public health. In order to save the cost and the time spent on identifying the functional sites in sequenced proteins in biology laboratory, computer programs have been widely used for decades. Many of them are implemented using the state-of-the-art pattern recognition algorithms, including decision trees, neural networks and support vector machines. Although the success of this effort has been obvious, advanced and new algorithms are still under development for addressing some difficult issues. This review will go through the major stages in developing pattern recognition algorithms for protein functional site prediction and outline the future research directions in this important area.


Regional collaboration in the Middle East to deal with H5N1 avian flu.


title: Single-tube single-enzyme reverse transcriptase PCR assay for detection of bovine viral diarrhea virus in pooled bovine serum.

text: A reverse transcriptase PCR (RT-PCR) was developed for use as a diagnostic screening test for the detection of bovine viral diarrhea virus (BVDV) in pooled bovine serum samples. Individual serum samples from 60 dairy cattle herds located in Pennsylvania were evaluated by the microplate virus isolation method, and pooled sera were analyzed by RT-PCR. RT-PCR was sensitive and specific and detected a single viremic serum sample in up to 100 pooled serum samples. RT-PCR analysis of pooled sera provides a rapid and cost-effective method for the screening of cattle herds for the presence of animals persistently infected with BVDV.


title: New structure model for the packaging signal in the genome of group IIa coronaviruses.

text: A 190-nucleotide (nt) packaging signal (PS) located in the 3' end of open reading frame 1b in the mouse hepatitis virus, a group IIa coronavirus, was previously postulated to direct genome RNA packaging. Based on phylogenetic data and structure probing, we have identified a 95-nt hairpin within the 190-nt PS domain which is conserved in all group IIa coronaviruses but not in the severe acute respiratory syndrome coronavirus (group IIb), group I coronaviruses, or group III coronaviruses. The hairpin is composed of six copies of a repeating structural subunit that consists of 2-nt bulges and 5-bp stems. We propose that repeating AA bulges are characteristic features of group IIa PSs.


title: Molecular mimicry between S peplomer proteins of coronaviruses (MHV, BCV, TGEV and IBV) and Fc receptor.

text: In previous studies we have demonstrated molecular mimicry between the S peplomer protein of Mouse Hepatitis Virus (MHV) and Fc gamma Receptor (Fc gamma R) of IgG. Rabbit IgG, but not its F(ab')2 fragments, monoclonal rat and mouse IgG and the rat 2.4G2 anti-mouse Fc gamma R monoclonal antibody (mab) immunoprecipitated natural and recombinant MHV S protein. On the basis of a number of criteria, MHV S peplomer protein exhibits Fc IgG binding ability. We report here a molecular mimicry between the S peplomer protein of Bovine Coronavirus (BCV) and Fc gamma R. BCV S peplomer protein which belongs to the same antigenic subgroup as MHV also binds Fc portion of rabbit IgG and is immunoprecipitated by the 2.4G2 anti-Fc gamma R mab. In contrast, Transmissible Gastroenteritis Coronavirus (TGEV) and Infectious Bronchitis Virus (IBV) S peplomer proteins which represent two distinct antigenic subgroups of Coronaviridae do not bind rabbit IgG and do not react with anti-Fc gamma R mab. However, homologous swine IgG, but not its F(ab')2 fragments, immunoprecipitated from TGEV-infected cells a polypeptide chain with molecular mass of 195 kDa, identical to that immunoprecipitated by the T36 mab anti-TGEV S peplomer protein.


title: Cerebral near-infrared spectroscopy in the care of patients during cardiological procedures: a summary of the clinical evidence.

text: Patients undergoing cardiological procedures generally have significant cardiovascular morbidity, and therefore these patients might be at risk for major periprocedural complications. The ability to closely monitor the hemodynamic status would present a major advantage to optimize patient care in this setting. The aim of this review is to assess the available evidence for the use of near-infrared spectroscopy (NIRS) in the care of patients during cardiological procedures. A systematic literature search was conducted using electronic bibliographic databases (MEDLINE, PubMed, EMBASE) from their first available date using the following search strategy: (spectroscopy, near-infrared OR infrared spectroscopy OR NIRS OR cerebral oxygen*) AND (interventional OR electrophysiological OR catheterization OR ablation OR ICD OR defibrillator). Inclusion criteria were limited to human studies, English language and cardiac patients. All manuscripts concerning the use of NIRS in every area of catheterization and electrophysiology were included. The literature search yielded eleven observational studies and five case reports concerning the research question. No randomized trials could be retrieved. Six studies evaluated NIRS during supraventricular and ventricular arrhythmias, one during transcatheter aortic valve implantations, and four studies assessed the use of NIRS in pediatric catheterization procedures. Overall, the studies demonstrated that NIRS provides a very quick representation of cerebral oxygen saturation and that it might identify changes that could not be predicted from standard hemodynamic monitoring. However, the evidence is currently too low to conclude that NIRS can optimize patient care during cardiological procedures.


title: Summary of notifiable diseases--United States, 2004.

text: The Summary of Notifiable Diseases--United States, 2004 contains the official statistics, in tabular and graphic form, for the reported occurrence of nationally notifiable infectious diseases in the United States for 2004. Unless otherwise noted, the data are final totals for 2004 reported as of December 2, 2005. These statistics are collected and compiled from reports sent by state health departments to the National Notifiable Diseases Surveillance System (NNDSS), which is operated by CDC in collaboration with the Council of State and Territorial Epidemiologists (CSTE). The Summary is available at http://www.cdc.gov/mmwr/summary.html. This site also includes publications from previous years.


title: Plant diversity patterns in neotropical dry forests and their conservation implications.

text: Seasonally dry tropical forests are distributed across Latin America and the Caribbean and are highly threatened, with less than 10% of their original extent remaining in many countries. Using 835 inventories covering 4660 species of woody plants, we show marked floristic turnover among inventories and regions, which may be higher than in other neotropical biomes, such as savanna. Such high floristic turnover indicates that numerous conservation areas across many countries will be needed to protect the full diversity of tropical dry forests. Our results provide a scientific framework within which national decision-makers can contextualize the floristic significance of their dry forest at a regional and continental scale.


title: Epidemiological characteristics of an outbreak of severe acute respiratory syndrome in Dongcheng District of Beijing from March to May 2003.

text: OBJECTIVE To describe epidemiologic features of an outbreak of severe acute respiratory syndrome (SARS) in Dongcheng District, Beijing occurred in a period between March and May 2003. METHODS Data of SARS cases notified from Dongcheng District Center for Disease Control and Prevention(CDC)and supplemented by other channels were collected. Clinicians and officials of local hospitals were interviewed in groups and medical records of fatal cases of SARS were reviewed to verify the diagnosis. Stored serum specimens of the patients were detected for IgG antibody against SARS Co-V by enzyme-linked immunosorbent assay (ELISA). All the data were input into dataset files by Microsoft Excel-2000 software and analyzed with SPSS version 10.0 software. RESULTS Outbreak of SARS in Dongcheng District started on March 14, 2003 with a peak in mid- and late April, and dropped in early May. A total of 572 reported cases were collected during this period in Dongcheng District, Beijing, and 99 of them were excluded from SARS, because of diagnosis of common cold, regular pneumonia, measles and rubella, etc. Actually, 473 SARS cases, which included 390 (82.5%) probable cases and 83 (17.5%) suspect cases, were analyzed. About 90% of the probable cases were positive for IgG antibody. Attack rate of SARS in permanent residents of Dongcheng District was 28.3 per 100 000. Forty-one of them died, with a case-fatality rate of 8.7%. Persons were all susceptible to infection of SARS Co-V, with the highest proportion at ages of 20-50 years, which accounted for 68.7% of the total cases. Average age of the patients at their onset was 40.7 years. No gender difference in SARS cases was found. Number of SARS cases in health-care workers (HCWs) accounted for 18.0% and that in retired workers accounted for 15.4% of the total cases. Cases occurred in all 10 sub-districts of Dongcheng, with the highest in Beixinqiao and Andingmen Sub-districts. Totally, 230 of the 572 notified cases (40.2%) were hospitalized at local hospitals under the jurisdiction of Dongcheng District. Eighteen of 85 cases of SARS occurred in HCWs of local hospitals, accounting for 4.5% of the total number of HCWs working at wards caring for SARS patients or fever clinics. There were 34.7% of SARS cases without any histories of contact before the onset of the disease. Familial aggregation phenomena were observed in 41.8% of the cases and 18.1% of households. And 7.4% (attack rate) of those exposed to SARS cases suffered from the illness during the periods of quarantine. CONCLUSIONS SARS appeared to be infectious in origin and caused outbreak in Dongcheng District, Beijing introduced by an imported case traveling from Hong Kong in a period between March and May 2003. People are all susceptible to infection of SARS Co-V, which mainly threatens the young adults and the middle-aged, as well as HCWs and the retired workers. The main mode of transmission is direct exposure to SARS patients in a near distance at hospitals or families via droplets spread. Prevention and control of SARS should be focused on early isolation of patients and quarantine for close contacts. Current available measures to prevent and control SARS are proved to be effective.


title: Analysis of the receptor-binding site of murine coronavirus spike protein.

text: It has been found that a domain composed of 330 amino acids of the N terminus of murine coronavirus spike protein [S1N(330)] is involved in receptor-binding activity (H. Kubo, Y.K. Yamada, and F. Taguchi, J. Virol. 68:5403-5410, 1994). To delineate the amino acid sequences involved in receptor-binding activity, we have compared the S1N(330) proteins of seven different mouse hepatitis virus MHV strains that are able to utilize the MHV receptor protein. Three conserved regions (sites I, II, and III) were found to consist of more than 10 identical amino acids, and they were analyzed for receptor-binding activity by site-directed mutagenesis. S1N(330) with a substitution at position 62 from the N terminus of S1 in region I and that with substitutions at positions 212, 214, and 216 in region II showed no receptor-binding activity. The S1N(330) mutants without receptor-binding activity were not able to prevent virus binding to the receptor. These results suggest that the receptor-binding site on S1N(330) is composed of regions located apart from each other in the protein's primary structure, in which Thr at position 62 as well as amino acids located at positions 212, 214, and 216 are particularly important.


title: The polypeptide structure of transmissible gastroenteritis virus.

text: The polypeptides of purified preparations of the coronavirus responsible for transmissible gastroenteritis of pigs have beem examined by polyacrylamide gel electrophoresis. Four major polypeptides, VPI (mol. wt. 200000), VP2 (50 000), VP3 (30000) and VP4 (28500) and two minor polypeptides, VPIa (105000) and VPIb (80500) have been reproducibly demonstrated in the virion, of which VPI, VP3 and VP4 contain carbohydrate. Treatment of the virion with the proteolytic enzyme bromelain removes the surface projections and VPI, thus identifying this glycopolypeptide as the major structural component of the projection.


title: Evaluation of body composition in COPD patients using multifrequency bioelectrical impedance analysis.

text: BACKGROUND Multifrequency bioelectrical impedance analysis (MF-BIA) is a technique that measures body impedance (Z) at different frequencies (5, 10, 50, 100, and 250 kHz). Body composition may be estimated using empirical equations, which include BIA variables or, alternatively, raw BIA data may provide direct information on water distribution and muscle quality. OBJECTIVES To compare raw MF-BIA data between COPD patients and controls and to study their relationship with respiratory and functional parameters in COPD patients. METHODS MF-BIA was performed (Human Im-Touch analyzer) in 212 COPD patients and 115 age- and BMI-matched controls. Fat-free mass (FFM) and fat mass were estimated from BIA data, and low- to high-frequency (5 kHz/250 kHz) impedance ratio was calculated. Physical fitness, lung function and respiratory muscle strength were also assessed in COPD patients. RESULTS After adjusting for age, weight, and body mass index, FFM and the 5/250 impedance ratio were lower in COPD patients (P<0.001) and were negatively affected by disease severity. In both male and female patients, the 5/250 impedance ratio was significantly correlated mainly with age (r=-0.316 and r=-0.346, respectively). Patients with a 5/250 impedance ratio below median value had lower handgrip strength (P<0.001), 6-minute walk distance (P<0.005), respiratory muscle strength (P<0.005), forced expiratory volume in 1 second (P<0.05) and vital capacity (P<0.005). Finally, the 5/250 impedance ratio was reduced (P<0.05) in patients with Global Initiative for Chronic Obstructive Lung Disease (GOLD) III and IV (compared to those with GOLD I and II) or a BODE index between 6 and 10 points (compared to those with BODE index between 1 and 5 points). CONCLUSION MF-BIA may be a useful tool for assessing body composition and nutritional status in COPD patients. In particular, the impedance ratio could give valuable information on cellular integrity and muscle quality.


docid: teyzripb

title: Fast, ultrasensitive virus detection using a Young interferometer sensor.

text: We report the application of an integrated optical Young interferometer sensor for ultrasensitive, real-time, direct detection of viruses. We have validated the sensor by detecting herpes simplex virus type 1 (HSV-1), but the principle is generally applicable. Detection of HSV-1 virus particles was performed by applying the virus sample onto a sensor surface coated with a specific antibody against HSV-1. The performance of the sensor was tested by monitoring virus samples at clinically relevant concentrations. We show that the Young interferometer sensor can specifically and sensitively detect HSV-1 at very low concentrations (850 particles/mL). We have further demonstrated that the sensor can specifically detect HSV-1 suspended in serum. Extrapolation of the results indicates that the sensitivity of the sensor approaches the detection of a single virus particle binding, yielding a sensor of unprecedented sensitivity with wide applications for viral diagnostics.


title: An unusual cause of ventricular tachycardia: Port-A-Cath fracture and embolization into the pulmonary artery.

text: We describe the case of a patient with a previously placed Port-A-Cath who was admitted to hospital for new onset of non-flushing catheter and palpitations with ventricular tachycardia. A chest X-ray and a linogram showed a Port-A-Cath fracture and distal embolization into the right ventricle resulting in ventricular tachycardia. The catheter was removed percutaneously using a Goose Neck snare with no complications and resolution of the ventricular tachycardia. The removed segment demonstrated thrombus. Prompt removal of the embolized catheter fragments should be undertaken given the subtle nature of the embolization and the potential complications.


title: SARS infection control in Taiwan: investigation of nurses' professional obligation.

text: This exploratory, cross-sectional, quantitative study investigated the relationship among hospital nurses' willingness to provide care for severe acute respiratory syndrome (SARS) patients, their attitudes toward SARS infection control measures, and nurses' health status and demographic characteristics. This project was conducted from May 6 to 12, 2003. A total of 126 nurses working in hospitals participated in this study. A conceptual model was developed, and the author designed a questionnaire to test this model. The developed model explained 32% of the variance in nurses' willingness to provide care for SARS patients. Nurses' levels of agreement with general SARS infection control measures, self-treatment of relief of fever and cough, necessity to close Hoping and Jenchi hospitals, nurses' physical health status, and holding a bachelor's degree were statistically significant predictors of nurses' willingness to care for SARS patients. Based on these findings, suggestions and study limitations are discussed.


title: Partial characterization of the principal soluble antigens associated with the coronavirus of transmissible gastroenteritis by complement fixation and immunodiffusion.

text: A microtiter complement fixation (CF) test to detect transmissible gastroenteritis (TGE) viral antigen was developed, using TGE hyperimmune pig serum as an antibody source. Sera from TGE covalescent pigs did not fix complement by this test. Maximal virus and soluble antigen (SA) titers were obtained 36 to 48 h after inoculation of swine testes cells. Cell-associated virus and SA titers were higher than those in the culture fluid, which had to be concentrated 20X before use as antigen in agar immunodiffusion tests (ID). By sucrose density-gradient centrifugation, the SA had a buoyant density of 1.10 g/ml and could be separated from the virus that banded in the 1.19-g/ml region. Virus and SA from three different isolates of TGE had the same buoyant densities. Heating and proteolytic enzyme digestion established the protein nature of the SA. As assayed by CF and ID, there were stability differences between crude and purified preparations of SA. Antibody prepared in rabbits against the SA neutralized the TGE virus.


title: Variations in disparate regions of the murine coronavirus spike protein impact the initiation of membrane fusion.

text: The prototype JHM strain of murine hepatitis virus (MHV) is an enveloped, RNA-containing coronavirus that has been selected in vivo for extreme neurovirulence. This virus encodes spike (S) glycoproteins that are extraordinarily effective mediators of intercellular membrane fusion, unique in their ability to initiate fusion even without prior interaction with the primary MHV receptor, a murine carcinoembryonic antigen-related cell adhesion molecule (CEACAM). In considering the possible role of this hyperactive membrane fusion activity in neurovirulence, we discovered that the growth of JHM in tissue culture selected for variants that had lost murine CEACAM-independent fusion activity. Among the collection of variants, mutations were identified in regions encoding both the receptor-binding (S1) and fusion-inducing (S2) subunits of the spike protein. Each mutation was separately introduced into cDNA encoding the prototype JHM spike, and the set of cDNAs was expressed using vaccinia virus vectors. The variant spikes were similar to that of JHM in their assembly into oligomers, their proteolysis into S1 and S2 cleavage products, their transport to cell surfaces, and their affinity for a soluble form of murine CEACAM. However, these tissue culture-adapted spikes were significantly stabilized as S1-S2 heteromers, and their entirely CEACAM-dependent fusion activity was delayed or reduced relative to prototype JHM spikes. The mutations that we have identified therefore point to regions of the S protein that specifically regulate the membrane fusion reaction. We suggest that cultured cells, unlike certain in vivo environments, select for S proteins with delayed, CEACAM-dependent fusion activities that may increase the likelihood of virus internalization prior to the irreversible uncoating process.


title: Thiopurine analogue inhibitors of severe acute respiratory syndrome-coronavirus papain-like protease, a deubiquitinating and deISGylating enzyme.

text: In the search for effective therapeutics against severe acute respiratory syndrome (SARS), 6-mercaptopurine (6MP) and 6-thioguanine (6TG) were found to be specific inhibitors for the SARS-coronavirus (CoV) papain-like protease (PLpro), a cysteine protease with deubiquitinating and deISGylating activity. 6MP and 6TG have long been used in cancer chemotherapy for treatment of acute lymphoblastic or myeloblastic leukaemia. Development and optimization of 6MP and 6TG will not only be important for antiviral studies, but also for further elucidating the biological functions of cellular deubiquitinating enzymes (DUBs) and deISGylating enzymes. So far, several crystal structures of cellular DUBs have been solved. Structure comparison has been carried out to search for DUBs with a similar structure to that of PLpro, and we have tried to dock 6MP and 6TG into these DUBs to investigate the potential use of 6MP and 6TG as cellular DUB inhibitors. The best docking score and binding energy for 6MP and 6TG is against ubiquitin-specific protease (USP)14, suggesting that 6MP and 6TG are potential inhibitors of USP14. Finding new usages for old drugs will speed up the process of drug discovery and substantially reduce the cost of drug development.


title: Hemagglutinin pseudotyped lentiviral particles: characterization of a new method for avian H5N1 influenza sero-diagnosis.

text: BACKGROUND Highly pathogenic avian influenza (HPAI) H5N1 has spread globally in birds and infected over 270 humans with an apparently high mortality rate. Serologic studies to determine the extent of asymptomatic H5N1 infection in humans and other mammals and to investigate the immunogenicity of current H5N1 vaccine candidates have been hampered by the biosafety requirements needed for H5N1 micro-neutralization tests. OBJECTIVE Development of a serodiagnostic tool for highly pathogenic influenza that reproduces H5N1 biology but can be used with less biohazard. STUDY DESIGN We have generated and evaluated H5 hemagglutinin pseudotyped lentiviral particles encoding the luciferase reporter (H5pp). RESULTS H5pp entry into target cells depends on alpha2-3 cell surface sialic acids and requires low pH for membrane fusion. H5pp infectivity is specifically neutralized by sera from patients and animals infected with H5N1 and correlates well with conventional microneutralization test. CONCLUSIONS H5pp reproduce H5N1 influenza virus entry into target cells and potentially provides a high-throughput and safe method for sero-epidemiology.


title: Role of viruses and atypical bacteria in asthma exacerbations among children in Oporto (Portugal).

text: BACKGROUND Upper respiratory tract infections are known to be a significant precipitant of acute asthma exacerbations. The aim of this study was to evaluate seasonal trends and the role of these pathogens in asthma exacerbations in school-aged children from Oporto (Portugal). METHODS Nasal aspirates were collected from children aged 6 to 12 years old with asthma exacerbations attended in the Pediatric Emergency Department one day per week from January 1 to December 31, 2003. Demographic data, severity of asthma and asthma exacerbations, and current treatment were recorded. Samples were obtained through nasal wash with 1 ml saline and were processed by immunofluorescence assays (respiratory syncytial virus, adenovirus, parainfluenza and influenza virus), retrotranscription polymerase chain reaction (rhinovirus) and polymerase chain reaction (enterovirus, Chlamydia pneumoniae and Mycoplasma pneumoniae). RESULTS In 54 eligible children, 37 nasal samples were obtained. Infectious agents were detected in 78 % of the patients. Rhinovirus was detected in 70.3 %, Mycoplasma pneumoniae in 16.2 %, enterovirus in 10.8 %, and Chlamydia pneumoniae in 2.7 %. Coinfection was identified in 21.6 % of the samples. There was no significant correlation between current treatment status, severity of asthma or exacerbations and the isolated agents. Two distinct peaks of asthma exacerbation were found, 40.5 % in spring and 32.4 % in autumn [corrected] The highest number of cases was recorded in March and the lowest in August and January. Rhinoviruses was detected in 27 % of the cases in autumn and in 24.3 % in spring [corrected] CONCLUSIONS These results confirm the previously reported high frequency of rhinovirus detection in asthma exacerbations in children and provide evidence that asthma exacerbations and rhinovirus infections follow a seasonal pattern, occurring mostly in spring and autumn. The findings also underscore the frequency of Mycoplasma pneumoniae detection, and emphasize the importance of this agent as a possible trigger of asthma exacerbations.


title: Specific epitopes of the structural and hypothetical proteins elicit variable humoral responses in SARS patients.

text: BACKGROUND Severe acute respiratory syndrome (SARS) is an infectious disease which was caused by a novel coronavirus (SARS-CoV). SARS has caused an outbreak in the world during 2003 and 2004, with 8098 individuals being infected and a death toll of 774 in 28 regions around the world. Specific humoral responses to viral infection remain unclear. OBJECTIVE To analyse the antigenicity of the SARS-CoV genome and identify potential antigenic epitopes in the structural proteins. METHODS Potential antigenic epitopes were identified in the structural proteins (nucleocapsid, membrane, spike, and small envelope proteins) and hypothetical proteins (SARS3a, 3b, 6, 7a, and 9b) that are specific for SARS-CoV. A peptide chip platform was created and the profiles of antibodies to these epitopes were investigated in 59 different SARS patients' sera obtained 6-103 days after the onset of the illness. Serial sera from five additional patients were also studied. RESULTS Epitopes at the N-terminus of the membrane protein and the C-terminus of nucleocapsid protein elicited strong antibody responses. Epitopes on the spike protein were only moderately immunogenic but the effects were persistent. Antibodies were also detected for some putative proteins, noticeably the C-termini of SARS3a and SARS6. CONCLUSIONS Important epitopes of the SARS-CoV genome that may serve as potential markers for the viral infection are identified. These specific antigenic sites may also be important for vaccine development against this new fatal infectious disease.


title: Effect of candesartan on microalbuminuria and albumin excretion rate in diabetes: three randomized trials.

text: BACKGROUND Microalbuminuria in diabetes is strongly predictive of nephropathy, end-stage renal disease, and premature cardiovascular morbidity and mortality. Effective preventive therapies are therefore a clinical priority. OBJECTIVE To determine whether the angiotensin-receptor blocker candesartan compared with placebo affects microalbuminuria incidence or rate of change in albuminuria in type 1 and type 2 diabetes. DESIGN 3 randomized trials of the DIRECT (Diabetic Retinopathy Candesartan Trials) Program. SETTING 309 secondary care centers. PATIENTS 3326 and 1905 patients with type 1 and type 2 diabetes, respectively. Most were normotensive, and all had normoalbuminuria (median urinary albumin excretion rate, 5.0 microg/min). INTERVENTION Candesartan, 16 mg/d increasing to 32 mg/d, versus placebo. Assignment was done centrally using an interactive voice-response system. Patients, caregivers, and researchers were blinded to treatment assignment. During a median follow-up of 4.7 years, 793 patients discontinued therapy and 63 were lost to follow-up. MEASUREMENTS Urinary albumin excretion rate, assessed annually by 2 overnight collections; if it was 20 microg/min or greater, then 2 further collections were done. The primary end point was new microalbuminuria (3 or 4 collections of urinary albumin excretion rate >or=20 microg/min). The secondary end point was rate of change in albuminuria. RESULTS Individual and pooled results of the 3 trials showed that candesartan had little effect on risk for microalbuminuria (pooled hazard ratio, 0.95 [95% CI, 0.78 to 1.16]; P = 0.60). Pooled results showed that the annual rate of change in albuminuria was 5.53% lower (CI, 0.73% to 10.14%; P = 0.024) with candesartan than with placebo. LIMITATIONS Investigators recruited mainly normotensive patients or patients with well-controlled hypertension who were at low overall vascular risk, which resulted in a low rate of microalbuminuria. Studies were powered for retinal and not renal end points. CONCLUSION Candesartan, 32 mg/d, for 4.7 years did not prevent microalbuminuria in mainly normotensive patients with type 1 or type 2 diabetes.


title: Abdominal Wall Reconstruction Using Retrorectus Self-adhering Mesh: A Novel Approach.

text: BACKGROUND In abdominal wall reconstruction, the retrorectus plane offers an ideal location for mesh placement. Mesh fixation in this plane is often achieved using transfascial sutures, which risks entrapping intercostal nerves and causing significant pain, and takes time to place. A novel alternative is the use of sutureless self-adhering mesh. Although the use of this mesh in inguinal hernias has been well described, studies on its use in abdominal wall reconstruction are lacking. METHODS Consecutive patients who underwent ventral hernia repair with retrorectus mesh were reviewed. This included patients who received transfascially sutured mesh and those who received sutureless self-adhering mesh. All patients were followed up for at least 12 months. The amount of narcotics required by each patient postoperatively was calculated. Surgical-site occurrences (SSOs) and hernia recurrence and bulge were measured. RESULTS Twenty-six patients underwent abdominal wall reconstruction with retrorectus mesh. This included 12 patients with transfascially sutured mesh and 14 patients with self-adhering mesh. Mean follow-up was 600 days. Baseline characteristics were similar between the 2 groups. Patients receiving self-adhering mesh required significantly less narcotics than patients with transfascially sutured mesh. There were no significant differences in the rate of SSOs between the 2 groups. No hernia recurrences, bulges, or chronic pain occurred in either group. CONCLUSIONS This is the first study to compare the outcomes of retrorectus self-adhering mesh and transfascially sutured mesh in abdominal wall reconstruction. Our results show low rates of SSO, recurrence, and bulge with both options, with significantly less acute pain with self-adhering mesh.


title: Daily Situation Report on Coronavirus disease (COVID-19) in Iran; March 25, 2020.

text: After detection of the first confirmed cases of COVID-19 in Iran, the National Committee on COVID-19 Epidemiology in Ministry of Health and Medical Education was established. This Committee is official source of gathering, analyzing, and reporting the COVID-19 data in Iran. The data of all sources in the country including, medical care monitoring center (MCMC), Hospitals' Information Systems (HIS), Laboratory portal, the data of the center for communicable disease control (MOH), as well as the data from community health centers are integrated and used in this regards. This factsheet contain daily situation report on coronavirus disease (covid-19) in Iran; March 25, 2020.


title: Avian astroviruses.

text: As poultry becomes more important in the world economy, it is increasingly important to fully understand the mechanisms of disease and poor production that affect the industry. To more accurately and reasonably treat these diseases, a more sophisticated understanding of interrelatedness is required. This review focuses on avian astroviruses (AAstVs), in particular the recent advances in our understanding of AAstV molecular biology, and also history, diagnosis, treatment and control. The known AastVs comprise duck astrovirus 1, turkey astrovirus 1 and 2, and avian nephritis virus of chickens. Nucleotide and amino acid identities between the avian and mammalian (human, ovine, bovine) astroviruses is very low (e.g. 20 to 25% and 12 to 15%, respectively) in open reading frame (ORF) 1a. There is also variation among the avian astroviruses, including between the two known types of turkey astrovirus. The ORF 1b sequence contains a number of conserved amino acid motifs; these could be the basis of degnerate oligonucleotide primers. A nomenclature for astroviruses is also proposed, based on: host species-astrovirus-type number/country(state)/reference number/year of isolation. For example, turkey astrovirus 2/North Carolina/034/1999.


title: Recombinant vesicular stomatitis virus vectors expressing herpes simplex virus type 2 gD elicit robust CD4+ Th1 immune responses and are protective in mouse and guinea pig models of vaginal challenge.

text: Recombinant vesicular stomatitis virus (rVSV) vectors offer an attractive approach for the induction of robust cellular and humoral immune responses directed against human pathogen target antigens. We evaluated rVSV vectors expressing full-length glycoprotein D (gD) from herpes simplex virus type 2 (HSV-2) in mice and guinea pigs for immunogenicity and protective efficacy against genital challenge with wild-type HSV-2. Robust Th1-polarized anti-gD immune responses were demonstrated in the murine model as measured by induction of gD-specific cytotoxic T lymphocytes and increased gamma interferon expression. The isotype makeup of the serum anti-gD immunoglobulin G (IgG) response was consistent with the presence of a Th1-CD4+ anti-gD response, characterized by a high IgG2a/IgG1 IgG subclass ratio. Functional anti-HSV-2 neutralizing serum antibody responses were readily demonstrated in both guinea pigs and mice that had been immunized with rVSV-gD vaccines. Furthermore, guinea pigs and mice were prophylactically protected from genital challenge with high doses of wild-type HSV-2. In addition, guinea pigs were highly protected against the establishment of latent infection as evidenced by low or absent HSV-2 genome copies in dorsal root ganglia after virus challenge. In summary, rVSV-gD vectors were successfully used to elicit potent anti-gD Th1-like cellular and humoral immune responses that were protective against HSV-2 disease in guinea pigs and mice.


title: Border screening for SARS in Australia: what has been learnt?

text: OBJECTIVE To assess the effectiveness of the Australian border entry screening program to detect arriving travellers with symptoms of severe acute respiratory syndrome (SARS). DESIGN AND SETTING Descriptive study of outcomes of screening at Australian airports and seaports between 5 April 2003 and 16 June 2003. To determine the number of international travellers who were symptomatic on arrival in Australia but missed by screening, data were obtained on the number of arrivals screened and the number with symptoms (from the Australian Quarantine and Inspection Service [AQIS]), as well as the number of people investigated for SARS (from the Australian SARS Case Register). RESULTS There were 1.84 million arrivals into Australia during the study period, and 794 were referred for screening to AQIS staff. Of these, the findings in four travellers were consistent with the World Health Organization case definition for SARS, and they were referred by the Chief Quarantine Officers to designated hospitals for further investigation. None of these people was confirmed to have SARS. One person reported as a probable SARS case acknowledged being symptomatic on arrival, but had been missed by border screening. CONCLUSIONS The low identification rate was attributed to the low prevalence of SARS, the use of exit screening by affected countries, and the subjective measures used in the screening process. With current knowledge about SARS, border screening should focus on educating incoming travellers, especially groups at high risk of transmitting the disease (the elderly and those with underlying chronic illnesses). Objective screening measures should be used during SARS outbreaks to prevent importation of the disease.


title: [Morphologic changes in the intestine of swine after infection with verotoxin-producing Escherichia coli].

text: Spontaneous morphologic lesions are described in 12 of 66 pigs submitted for necropsy. All 12 pigs were culture positive for Verotoxin-producing E. coli (VTEC). 10 of them were weaned pigs, one a suckling piglet and one a fattening hog. In 6 cases E. coli serovar 0139:H1 and in one case each the serovars 0139:H40; 0138:H-; 0125ac:H27 and 0154:H- were isolated. From the fecal samples of 2 animals E. coli ONT (O-group non typable):H- were cultured. Macroscopically there were cyanosis, edema of the eye lids, catarrhal enteritis and/or colitis as well as edema of the mesentery, swelling of the mesenteric lymph nodes and congestion of the lung to varying degrees. Histopathologic examination of 5 animals was carried out. In 3 animals atrophy and edema of the villi in the jejunum and ileum were discovered. In one animal an additional infection with corona virus was confirmed electron microscopically. Furthermore there was disseminated necrosis of lymphocytes in Peyer's patches of the small intestine and in secondary follicles of the mesenteric lymph nodes. In one of the animals a hemorrhagic-necrotising ileitis, occurred characterized by necrosis of villi and thrombosis of blood vessels in the mucosa. The highest number of VTEC with seven out of twelve animals was found in weaned pigs in association with the E. coli serovar 0139.


Re-emerging diseases and epidemic threats: Ten 'areas of actions' (AA-10) and a case for global solidarity.


title: High coverage of influenza vaccination among healthcare workers can be achieved during heightened awareness of impending threat.

text: INTRODUCTION As preparation against a possible avian flu pandemic, international and local health authorities have recommended seasonal influenza vaccination for all healthcare workers at geographical risk. This strategy not only reduces "background noise", but also chance of genetic shifts in avian influenza viruses when co-infection occurs. We evaluate the response of healthcare workers, stratified by professional groups, to a non-compulsory annual vaccination call, and make international comparisons with countries not at geographical risk. MATERIALS AND METHODS A cross-sectional study was performed over the window period for vaccination for the 2004 to 2005 influenza season (northern hemisphere winter). The study population included all adult healthcare workers (aged < or =21 years) employed by a large acute care tertiary hospital. RESULTS The uptake rates among frontline caregivers--doctors >50%, nurses >65% and ancillary staff >70%--markedly exceeded many of our international counterparts results. CONCLUSION Given its close proximity in time and space to the avian flu pandemic threat, Singapore healthcare workers responded seriously and positively to calls for preventive measures. Other factors, such as the removal of financial, physical and mental barriers, may have played important facilitative roles as well.


title: Translation and processing of mouse hepatitis virus virion RNA in a cell-free system.

text: The first event after infection with mouse hepatitis virus strain A59 (MHV-A59) is presumed to be the synthesis of an RNA-dependent RNA polymerase from the input genomic RNA. The synthesis and processing of this putative polymerase protein was studied in a cell-free translation system utilizing 60S RNA from MHV-A59 virions. The polypeptide products of this reaction included two major species of 220 and 28 kilodaltons. Kinetics experiments indicated that both p220 and p28 appeared after 60 min of incubation and that protein p28 was synthesized initially as the N-terminal portion of a larger precursor protein. When the cell-free translation products were labeled with N-formyl[35S]methionyl-tRNAi, p28 was the predominant radioactive product, confirming its N-terminal location within a precursor protein. Translation in the presence of the protease inhibitors leupeptin and ZnCl2 resulted in the disappearance of p28 and p220 and the appearance of a new protein, p250. This product, which approached the maximal size predicted for a protein synthesized from genomic RNA, was not routinely detected in the absence of inhibitors even under conditions which optimized the translation reaction for elongation of proteins. Subsequent chelation of ZnCl2 resulted in the partial cleavage of the precursor protein and the reappearance of p28. One-dimensional peptide mapping with Staphylococcus aureus V-8 protease confirmed the precursor-product relationship of p250 and p28. The results show that MHV virion RNA, like many other viral RNAs, is translated into a large polyprotein, which is cleaved soon after synthesis into smaller, presumably functional proteins. This is in marked contrast to the synthesis of other MHV proteins, in which minimal proteolytic processing occurs.


title: Molecular evolution of the SARS coronavirus during the course of the SARS epidemic in China.

text: Sixty-one SARS coronavirus genomic sequences derived from the early, middle, and late phases of the severe acute respiratory syndrome (SARS) epidemic were analyzed together with two viral sequences from palm civets. Genotypes characteristic of each phase were discovered, and the earliest genotypes were similar to the animal SARS-like coronaviruses. Major deletions were observed in the Orf8 region of the genome, both at the start and the end of the epidemic. The neutral mutation rate of the viral genome was constant but the amino acid substitution rate of the coding sequences slowed during the course of the epidemic. The spike protein showed the strongest initial responses to positive selection pressures, followed by subsequent purifying selection and eventual stabilization.


title: Detection of animal viruses using nucleic acid sequence-based amplification (NASBA).

text: As seen in recent avian influenza outbreaks in Asia, prevention is the key to fighting infectious disease successfully. Efficient disease surveillance systems on the basis of molecular diagnostics will help monitor the emergence of viruses in the early stage and thus prompt containment measures can be in place to minimize disease spread. Here we describe and review molecular diagnostics focusing on nucleic acid sequence-based amplification (NASBA) technology in detecting viruses causing animal diseases, such as avian influenza, foot-and-mouth disease, and Newcastle disease. NASBA offers high sensitivity, specificity, accuracy, and speed of availability of results, and NASBA would be the most applicable molecular diagnostics for disease surveillance and control.


title: Ethical issues in resource triage.

text: Mass-care events, such as pandemic influenza, could reach such devastating proportions that there will be the need to make difficult triage decisions that will ultimately result in the deaths or severe disability of patients in large numbers. The method by which we determine how triage of scarce health care resources will be performed must be clearly defined prior to a disaster event. This paper will discuss several of the ethical principles that must be weighed in developing a mass-care triage plan, as well as steps to facilitate its implementation. Development of triage policies in such an event should be developed in an open and transparent manner, be reasonable in design, include the views of the critical stakeholders, and be responsive to and provide a mechanism for accountability, with a clearly defined goal of the just triage of limited health care resources. Planning failure will result in increased deaths from poor triage processes and substantial mistrust of the health care system and its practitioners.


Public health law and ethics: lessons from SARS and quarantine.


title: [New coronavirus infection: new challenges, new legacies].

text: BACKGROUND A new highly pathogenic human coronavirus has emerged. Its natural history and its determinants are still under investigation. It lacks a publication to examine all the cases identified worldwide. Therefore, the objective of this paper is to describe the cases and deaths caused by the new coronavirus. METHODS World Health Organization's, European Centre for Disease Control's and Eurosurveillance's publications online were reviewed. We performed a descriptive analysis of the cases; the limits of proportions were calculated with a 0.05 alpha using Wilson test, and Student's t test for mean difference. RESULTS There are 17 confirmed cases and 11 deaths in several countries in Asia and Europe; men predominated. The case fatality rate was 64.70 %. People who died took five days to hospitalize after the first symptoms. There is a lack of publications to describe the natural history of the disease, but the descriptions of the European publications are consistent with the results of this study. CONCLUSION It is necessary to keep the surveillance and further studies in order to assess the impact on international public health.


title: Assessment of Stomoxys calcitrans (Diptera: Muscidae) as a vector of porcine reproductive and respiratory syndrome virus.

text: Porcine Reproductive and respiratory syndrome (PRRS) is a globally significant swine disease caused by an arterivirus. The virus replicates in alveolar macrophages of infected pigs, resulting in pneumonia in growing pigs and late-term abortions in sows. Outbreaks occur on disparate farms within an area despite biosecurity measures, suggesting mechanical transport by arthropods. We investigated the vector potential of stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), in the transmission of porcine reproductive and respiratory syndrome virus (family Arteriviridae, genus Arterivirus, PRRSV) under laboratory conditions. Stable flies were collected around PRRS-negative boar stud barns in North Carolina and tested for presence of the virus. Stable flies were collected on alsynite traps placed near the exhaust fan of the close-sided tunnel-ventilated buildings, suggesting blood seeking flies are attracted by olfactory cues. No flies were positive for PRRSV. We assessed transmission of the virus through an infective bite by feeding laboratory reared stable flies on blood containing virus and transferring them to naive pigs for subsequent bloodmeals. Transmission of the virus to naive pigs by infective bites failed in all attempts. The volume of blood contained within the closed mouthparts of the stable fly seems to be insufficient to deliver an infective dose of the virus. Stable flies are unlikely to transmit PRRSV from one pig to another while blood feeding. The fate of the virus after a bloodmeal remains to be determined.


title: Risk perception and compliance with quarantine during the SARS outbreak.

text: PURPOSE To explore the experience of being on quarantine for severe acute respiratory syndrome (SARS) with a focus on the relationship between perceived risk of contracting SARS and reported compliance with the quarantine order and protocols. DESIGN Descriptive, qualitative. METHODS Semi-structured interviews were conducted with people who had been quarantined during the SARS outbreak in Toronto in 2003. Data analysis was completed using an iterative and collaborative approach of reading and re-reading the transcribed interviews, identifying common themes, and comparing and contrasting the data. FINDINGS To varying extents, participants wavered between fear and denial about their risk of contracting or spreading SARS. Reported compliance with the actual quarantine order was high. However, within households quarantine protocols were followed unevenly. CONCLUSIONS This research indicates the need for greater credibility in public health communications to increase compliance with quarantine protocols and to contain outbreaks of new and deadly infectious diseases.


Evaluating and optimizing outcomes of surgery for endocarditis.


title: Complete genome structure and phylogenetic analysis of little cherry virus, a mealybug-transmissible closterovirus.

text: The 5'-terminal genomic region (8597 nt) of little cherry virus (LChV), a mealybug-borne closterovirus, was cloned from double-stranded RNA, and its sequence determined to complete the 16934 nt sequence of the monopartite LChV RNA genome. In the 5' to 3' direction, the sequence encompasses ORF 1a, encoding the conserved replicative domains of methyltransferase and helicase, and ORF 1b, encoding RNA polymerase. ORFs 1a and 1b partially overlap (in O/+1 configuration), and the LChV replicase is probably expressed by ribosomal frameshifting as a fusion product with a molecular mass of 318 kDa. The N-terminal part of the ORF 1a product contains a papain-like cysteine proteinase (PCP) domain with a predicted cleavage site between Gly-619 and Ser-620. The PCP and the upstream protein domains can be aligned with the equivalent parts of the leader proteins encoded by the whitefly-transmitted lettuce infectious yellows and sweet potato sunken vein closteroviruses. Phylogenetic reconstruction based on the aligned RNA polymerase sequences clearly suggests that the aphid-transmissible and whitefly-transmissible closteroviruses represent two distinct evolutionary lineages, with the mealybug-transmissible LChV being the most remote member of the 'whitefly' lineage.


title: Role of cyclinD1 and CDK4 in the carcinogenesis induced by silica.

text: OBJECTIVE To study the role of cyclinD1 and CDK4 in malignant transformation of human fetal lung diploid fibroblast cell line (2BS) induced by silica. METHODS Recombination vectors with sense and antisense pXJ41-cyclinD1 and pXJ41-CDK4 were constructed, and then transfected into the malignant transformed cells induced by silica, respectively. At the same time, pXJ41-neo was used as the control. RESULTS During the progress of the malignant transformation of 2BS cells induced by silica, cyclinD1 and CDK4 were overexpressed. Antisense RNA suppressed cyclinD1 and CDK4 gene expression in the antisense pXJ41-cyclinD1 and pXJ41-CDK4 transfected cells. Antisense RNA led to cell cycle arrest, resulting in lengthened G1 phase (the percentages of cells in the G1 phase changed from 45.1% to 52.7% and 58.0% for cyclinD1 and CDK4 transfected cells, respectively), and eventually attenuated the increase of the proliferation of malignant transformed cells induced by silica. Compared with malignant transformed cells induced by silica, cells transfected with antisense pXJ41-cyclinD1 and pXJ41-CDK4 showed obviously reduced growth rates. On the 8th day, the suppression rates were 58.69 and 77.43% (the growth rate of malignant transformed cells induced by silica was 100%), doubling time changed from 21.0 h to 31.4 h and 21.0 h to 42.7 h, respectively, the growth capacities on soft agar of cells transfected by antisense pXJ41-cyclinD1 and pXJ41-CDK4 decreased obviously. CONCLUSION CyclinD1 and CDK4 play an important role in maintaining transformed phenotype of the cancer cells.


title: Assembly of the coronavirus envelope: homotypic interactions between the M proteins.

text: The viral membrane proteins M and E are the minimal requirements for the budding of coronavirus particles. Since the E protein occurs in particles only in trace amounts, the lateral interactions between the M proteins apparently generate the major driving force for envelope formation. By using coimmunoprecipitation and envelope incorporation assays, we provide extensive evidence for the existence of such M-M interactions. In addition, we determined which domains of the M protein are involved in this homotypic association, using a mutagenetic approach. Mutant M proteins which were not able to assemble into viruslike particles (VLPs) by themselves (C. A. M. de Haan, L. Kuo, P. S. Masters, H. Vennema, and P. J. M. Rottier, J. Virol. 72:6838-6850, 1998) were tested for the ability to associate with other M proteins and to be rescued into VLPs formed by assembly-competent M proteins. We found that M proteins lacking parts of the transmembrane cluster, of the amphipathic domain, or of the hydrophilic carboxy-terminal tail, or M proteins that had their luminal domain replaced by heterologous ectodomains, were still able to associate with assembly-competent M proteins, resulting in their coincorporation into VLPs. Only a mutant M protein in which all three transmembrane domains had been replaced lost this ability. The results indicate that M protein molecules interact with each other through multiple contact sites, particularly at the transmembrane level. Finally, we tested the stringency with which membrane proteins are selected for incorporation into the coronavirus envelope by probing the coassembly of some foreign proteins. The observed efficient exclusion from budding of the vesicular stomatitis virus G protein and the equine arteritis virus M protein indicates that envelope assembly is indeed a highly selective sorting process. The low but detectable incorporation of CD8 molecules, however, demonstrated that this process is not perfect.


title: Detection of respiratory viruses and subtype identification of influenza A viruses by GreeneChipResp oligonucleotide microarray.

text: Acute respiratory infections are significant causes of morbidity, mortality, and economic burden worldwide. An accurate, early differential diagnosis may alter individual clinical management as well as facilitate the recognition of outbreaks that have implications for public health. Here we report on the establishment and validation of a comprehensive and sensitive microarray system for detection of respiratory viruses and subtyping of influenza viruses in clinical materials. Implementation of a set of influenza virus enrichment primers facilitated subtyping of influenza A viruses through the differential recognition of hemagglutinins 1 through 16 and neuraminidases 1 through 9. Twenty-one different respiratory virus species were accurately characterized, including a recently identified novel genetic clade of rhinovirus.


title: Case fatality rate of severe acute respiratory syndromes in Beijing.

text: OBJECTIVE To describe the case fatality rate of SARS in Beijing. METHODS Data of SARS cases notified from Beijing Center for Disease Control and Prevention (BCDC) and supplemented by other channels were collected. The data were analyzed by rate calculation. RESULTS The case fatality rate of SARS in Beijing was 7.66%, and had an ascending trend while the age of cases was getting older, and a descending trend while the epidemic development. The case fatality rate in Beijing was lower than that in other main epidemic countries or regions. CONCLUSIONS The risk of death increases with the increment of age of SARS patients. Beijing is successful in controlling and treating SARS.


title: The impact of chronic conditions on the economic burden of cancer survivorship: a systematic review.

text: INTRODUCTION This systematic review examines the excess cost of chronic conditions on the economic burden of cancer survivorship among adults in the US. Areas covered: Twelve published studies were identified. Although studies varied substantially in populations, comorbidities examined, methods, and types of cost reported, costs for cancer survivors with comorbidities generally increased with greater numbers of comorbidities or an increase in comorbidity index score. Survivors with comorbidities incurred significantly more in total medical costs, out-of-pocket costs, and costs by service type compared to cancer survivors without additional comorbidities. Expert commentary: Cancer survivors with comorbidities bear significant excess out-of-pocket costs and their care is also more expensive to the healthcare system. On-going evaluation of different payment models, care coordination, and disease management programs for cancer survivors with comorbidities will be important in monitoring impact on healthcare costs.


What's new in the pathogenesis of multiple sclerosis? A review.


title: Association of cytokine and chemokine gene polymorphisms with severe acute respiratory syndrome.

text: 1. The IFN-gamma +874A allele and RANTES -28 G allele are risk factors for SARS susceptibility. 2. The RANTES -28 G allele plays a role in the pathogenesis of SARS. 3. The polymorphisms of IL-10, TNF-alpha, IL-12, IP-10, Mig and MCP-1 are not associated with SARS susceptibility.


title: Clinical interventions for severe and critical COVID-19: what are the options.

text: The coronavirus disease 2019 (COVID-19) has been ongoing outbreak and declared as a global public health emergency by the World Health Organization. Severe and critical COVID-19 has high fatality rate due to complications like acute respiratory distress syndrome, acute respiratory failure or multiple organ failure. So far, there have been mounting research on the epidemiological and clinical characteristics of COVID-19. However, the information regarding treatment of severe and critical COVID-19 is limited. The current study reviewed published evidence of clinical interventions of severe and critical COVID-19, aiming to provide an up-to-date reference for further clinical treatment.


title: [Use of glucocorticoid in treatment of severe acute respiratory syndrome cases].

text: OBJECTIVE To explore the effect of glucocorticoid in the treatment for severe acute respiratory syndrome (SARS). METHODS Clinical data of 70 patients with SARS admitted to Youan Hospital in Beijing during March to May, 2003 were analyzed. RESULTS (1) Sixty-three of 70 cases of SARS recovered and seven cases died, with a case-fatality ratio of 10%. (2) Average length of hospital stay was 16.9 days for the all 70 cases, and 16.8 days for the 11 cases without glucocorticoid treatment, without statistical significance (F = 1.018, P = 0.39). (3) The other 59 cases were administered with 40 mg to 640 mg of methylprednisolone daily. (4) Average hospital stay was 15 days for the 23 cases with lower dose of 40 mg to 80 mg methylprednisolone daily, 18.5 days for the 27 cases with medium dose of 120 mg to 240 mg daily, and 17.9 days for the nine cases with higher dose of 320 mg to 640 mg daily (F = 1.018, P = 0.39). CONCLUSIONS Earlier use of glucocorticoid therapy with suitable doses could alleviate their clinical symptoms, improve their clinical courses, and promote the absorbance of infiltration in their lungs on chest-X-ray films for the cases with SARS. However, current clinical data showed that glucocorticoid therapy could not shorten the length of hospitalization for the cases with SARS.


Hope in a global pandemic.


title: Systematic assembly of a full-length infectious cDNA of mouse hepatitis virus strain A59.

text: A novel method was developed to assemble a full-length infectious cDNA of the group II coronavirus mouse hepatitis virus strain A59 (MHV-A59). Seven contiguous cDNA clones that spanned the 31.5-kb MHV genome were isolated. The ends of the cDNAs were engineered with unique junctions and assembled with only the adjacent cDNA subclones, resulting in an intact MHV-A59 cDNA construct of approximately 31.5 kb in length. The interconnecting restriction site junctions that are located at the ends of each cDNA are systematically removed during the assembly of the complete full-length cDNA product, allowing reassembly without the introduction of nucleotide changes. RNA transcripts derived from the full-length MHV-A59 construct were infectious, although transfection frequencies were enhanced 10- to 15-fold in the presence of transcripts encoding the nucleocapsid protein N. Plaque-purified virus derived from the infectious construct replicated efficiently and displayed similar growth kinetics, plaque morphology, and cytopathology in murine cells as did wild-type MHV-A59. Molecularly cloned viruses recognized the MHV receptor (MHVR) for docking and entry, and pretreatment of cells with monoclonal antibodies against MHVR blocked virus entry and replication. Cells infected with molecularly cloned MHV-A59 virus expressed replicase (gene 1) proteins identical to those of laboratory MHV-A59. Importantly, the molecularly cloned viruses contained three marker mutations that had been derived from the engineered component clones. Full-length infectious constructs of MHV-A59 will permit genetic modifications of the entire coronavirus genome, particularly in the replicase gene. The method has the potential to be used to construct viral, microbial, or eukaryotic genomes approaching several million base pairs in length and used to insert restriction sites at any given nucleotide in a microbial genome.


title: Photonic Shell-Crosslinked Nanoparticle Probes for Optical Imaging and Monitoring.

text: A pH-insensitive fluorophore is made to give pH-driven responses through its covalent incorporation within a nanostructure derived from pH-responsive polymers. Fluorophore-shell-crosslinked nanoparticles (SCKs) demonstrate notable enhancement of photophysical properties, in the physiological pH region. Fluorophore-SCKs are designed to swell at higher pH and shrink as the pH is lowered, producing high fluorescence vs. low fluorescence outputs, respectively.


A walk on the wild side--emerging wildlife diseases.


title: An outbreak of canine coronavirus in puppies in a Greek kennel.

text: Canine coronavirus (CCoV) is usually the cause of mild gastroenteritis in dogs and is known to have spread worldwide. However, to date, no CCoV cases have been confirmed in Greece. In the present work, the authors investigated an outbreak of enteritis in puppies from a Greek kennel for the presence of CCoV. Dogs were presented with clinical signs of diarrhea, anorexia, weakness, depression, dehydration, and 1 death. Canine coronavirus type II was detected by reverse transcription nested polymerase chain reaction in all 11 puppies, whereas 1 puppy presented dual infection with CCoV type II and canine parvovirus 2. Surprisingly, sequence analysis of the samples revealed higher similarity to the pantropic CCoV II strain CB/05 than to other reference strains, in the most variable region of the S gene.


title: Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: activation of cell-fusing activity of virions by trypsin and separation of two different 90K cleavage fragments.

text: In the murine coronavirus mouse hepatitis virus, a single glycoprotein, E2, is required both for attachment to cells and for cell fusion. Cell fusion induced by infection with mouse hepatitis virus strain A59 was inhibited by the addition of monospecific anti-E2 antibody after virus adsorption and penetration. Adsorption of concentrated coronavirions to uninfected cells did not cause cell fusion in the presence of cycloheximide. Thus, cell fusion was induced by E2 on the plasma membrane of infected 17 Cl 1 cells but not by E2 on virions grown in these cells. Trypsin treatment of virions purified from 17 Cl 1 cells quantitatively cleaved 180K E2 to 90K E2 and activated cell-fusing activity of the virions. This proteolytic cleavage yielded two different 90K species which were separable by sodium dodecyl sulfate-hydroxyapatite chromatography. One of the trypsin cleavage products, 90A, was acylated and may be associated with the lipid bilayer. The other, 90B, was not acylated and yielded different peptides than did 90A upon limited digestion with thermolysin or staphylococcal V8 protease. Thus, the cell-fusing activity of a coronavirus required proteolytic cleavage of the E2 glycoprotein, either by the addition of a protease to virions or by cellular proteases acting on E2, which was transported to the plasma membrane during virus maturation. There is a striking functional similarity between the E2 glycoprotein of coronavirus, which is a positive-strand RNA virus, and the hemagglutinin glycoprotein of negative-strand orthomyxoviruses, in that a single glycoprotein has both attachment and protease-activated cell-fusing activities.


title: In situ delivery of passive immunity by lactobacilli producing single-chain antibodies.

text: Lactobacilli have previously been used to deliver vaccine components for active immunization in vivo. Vectors encoding a single-chain Fv (scFv) antibody fragment, which recognizes the streptococcal antigen I/II (SAI/II) adhesion molecule of Streptococcus mutans, were constructed and expressed in Lactobacillus zeae (American Type Culture Collection (ATCC) 393). The scFv antibody fragments secreted into the supernatant or expressed on the surface of the bacteria showed binding activity against SAI/II in enzyme-linked immunosorbent assay (ELISA), and surface scFv-expressing lactobacilli agglutinated SAI/II-expressing S. mutans in vitro without affecting the corresponding SAI/II knockout strain. Lactobacilli expressing the scFv fragment fused to an E-tag were visualized by scanning electron microscopy (SEM) using beads coated with a monoclonal anti-E-tag antibody, and they bound directly to beads coated with SAI/II. After administration of scFv-expressing bacteria to a rat model of dental caries development, S. mutans bacteria counts and caries scores were markedly reduced. As lactobacilli are generally regarded as safe (GRAS) microorganisms, this approach may be of considerable commercial interest for in vivo immunotherapy.


Making dollars and sense of antisense.


title: Establishment of a fluorescent polymerase chain reaction method for the detection of the SARS-associated coronavirus and its clinical application.

text: OBJECTIVE To establish a fluorescent polymerase chain reaction (F-PCR) method for detecting the coronavirus related to severe acute respiratory syndrome (SARS) and to evaluate its value for clinical application. METHODS The primers and the fluorescence-labeled probe were designed and synthesized according to the published sequence of the SARS-associated coronavirus genes. A F-PCR diagnosis kit for detecting the coronavirus was developed, and 115 clinical nasopharyngeal gargling liquid samples were tested. RESULTS The sequence of PCR amplified products completely matched the related sequence of the SARS-associated coronavirus genome. Forty-nine out of 67 samples from identified SARS patients and 8 of 18 samples from persons having close contact with SARS patients showed positive results. All 30 samples from healthy controls were negative. CONCLUSION The F-PCR method established may be a rapid, accurate and efficient way for screening and for the early diagnosis of SARS patients.