title: Contemporary surgical management of advanced end stage emphysema: an evidence based review.

text: Emphysema is a progressive unrelenting component of chronic obstructive pulmonary disease and a major source of mortality and morbidity globally. The prevalence of moderate to severe emphysema is approximately 5% in Malaysia and likely to increase in the future. Hence advanced emphysema will emerge as a leading cause of hospital admission and a major consumer of healthcare resources in this country in the future. Patients with advanced disease have a poor quality of life and reduced survival. Medical therapy has been largely ineffective for many patients however certain subgroups have disease amenable to surgical palliation. Effective surgical therapies include lung volume reduction surgery, lung transplantation and bullectomy. This article is a comprehensive evidence based review of the literature evaluating the rationale, efficacy, safety and limitations of surgery for advanced emphysema highlighting the importance of meticulous patient selection and local factors relevant to Malaysia.


title: [Synthesis and activity of ACE inhibitory peptides].

text: To find anti-hypertensive lead drug, angiotensin converting enzyme (ACE) inhibitory peptides were synthesized and their effects on inhibiting ACE activity were investigated. ACE inhibitory peptides were synthesized via Fmoc solid-phase synthesis, isolated and purified through reversed phase high-performance liquid chromatography (RP-HPLC), and identified by mass spectrometry. A RP-HPLC analysis method was used to test ACE inhibitory activity in vitro of these ACE inhibitory peptides. Six octapeptides were successfully synthesized, and the analytical results of mass spectrum were consistent with their theoretically calculated data. Among these synthetic octapeptides, the anti-SARS (severe acute respiratory syndromes) octapeptide had the most obvious ACE inhibitory activity with an IC50 value of 3.4 x 10(-5) mol x L(-1). So octapeptide AVLQSGFR-OH (anti-SARS peptide) was found to be the strongest candidate for potential development as an anti-hypertensive drug and had the implication of further study.


docid: eatgrddn

title: Functional importance of the coiled-coil of the Ebola virus glycoprotein.

text: Ebola virus contains a single glycoprotein (GP) that is responsible for receptor binding and membrane fusion and is proteolytically cleaved into disulfide-linked GP1 and GP2 subunits. The GP2 subunit possesses a coiled-coil motif, which plays an important role in the oligomerization and fusion activity of other viral GPs. To determine the functional significance of the coiled-coil motif of GP2, we examined the effects of peptides corresponding to the coiled-coil motif of GP2 on the infectivity of a mutant vesicular stomatitis virus (lacking the receptor-binding/fusion protein) pseudotyped with the Ebola virus GP. A peptide corresponding to the C-terminal helix reduced the infectivity of the pseudotyped virus. We next introduced alanine substitutions into hydrophobic residues in the coiled-coil motif to identify residues important for GP function. None of the substitutions affected GP oligomerization, but some mutations, two in the N-terminal helix and all in the C-terminal helix, reduced the ability of GP to confer infectivity to the mutant vesicular stomatitis virus without affecting the transport of GP to the cell surface, its incorporation into virions, and the production of virus particles. These results indicate that the coiled-coil motif of GP2 plays an important role in facilitating the entry of Ebola virus into host cells and that peptides corresponding to this region could act as efficient antiviral agents.


title: Coronavirus transcription: subgenomic mouse hepatitis virus replicative intermediates function in RNA synthesis.

text: Both genomic and subgenomic replicative intermediates (RIs) and replicative-form (RF) structures were found in 17CL1 mouse cells that had been infected with the A59 strain of mouse hepatitis virus (MHV), a prototypic coronavirus. Seven species of RNase-resistant RF RNAs, whose sizes were consistent with the fact that each was derived from an RI that was engaged in the synthesis of one of the seven MHV positive-strand RNAs, were produced by treatment with RNase A. Because the radiolabeling of the seven RF RNAs was proportional to that of the corresponding seven positive-strand RNAs, the relative rate of synthesis of each of the MHV positive-strand RNAs may be controlled by the relative number of each of the size classes of RIs that are produced. In contrast to alphavirus, which produced its subgenome-length RF RNAs from genome-length RIs, MHV RF RNAs were derived from genome- and subgenome-length RIs. Only the three largest MHV RF RNAs (RFI, RFII, and RFIII) were derived from the RIs that migrated slowest on agarose gels. The four smallest RF RNAs (RFIV, RFV, RFVI, and RFVII) were derived from RIs that migrated in a broad region of the gel that extended from the position of 28S rRNA to the position of the viral single-stranded MHV mRNA-3. Because all seven RIs were labeled during very short pulses with [3H]uridine, we concluded that the subgenome-length RIs are transcriptionally active. These findings, with the recent report of the presence of subgenome-length negative-strand RNAs in cells infected with porcine transmissible gastroenteritis virus (P. B. Sethna, S.-L. Hung, and D. A. Brian, Proc. Natl. Acad. Sci. USA 86: 5626-5630, 1989), strongly suggest that coronaviruses utilize a novel replication strategy that employs the synthesis of subgenomic negative strands to produce subgenomic mRNAs.


docid: ahevwrgf

title: Modelling disease spread and control in networks: implications for plant sciences.

text: Networks are ubiquitous in natural, technological and social systems. They are of increasing relevance for improved understanding and control of infectious diseases of plants, animals and humans, given the interconnectedness of today's world. Recent modelling work on disease development in complex networks shows: the relative rapidity of pathogen spread in scale-free compared with random networks, unless there is high local clustering; the theoretical absence of an epidemic threshold in scale-free networks of infinite size, which implies that diseases with low infection rates can spread in them, but the emergence of a threshold when realistic features are added to networks (e.g. finite size, household structure or deactivation of links); and the influence on epidemic dynamics of asymmetrical interactions. Models suggest that control of pathogens spreading in scale-free networks should focus on highly connected individuals rather than on mass random immunization. A growing number of empirical applications of network theory in human medicine and animal disease ecology confirm the potential of the approach, and suggest that network thinking could also benefit plant epidemiology and forest pathology, particularly in human-modified pathosystems linked by commercial transport of plant and disease propagules. Potential consequences for the study and management of plant and tree diseases are discussed.


Structural and antigenic relationships between human, murine and avian coronaviruses.


title: Geriatric infections: Decreased immunity or evolved opportunists?

text: In host-parasite co-evolution, parasites are assumed to have an advantage owing to their shorter generation time. Evolution of pathogens within the lifetime of a host individual is implicated as a strong selective force in the evolution of sex and aging in the host. However, this assumption or its testable predictions have not been examined empirically. We classified infectious bacteria and viruses into those that can have continued longterm existence on the host body (group 1) versus those that have only a short-term interaction during an active infection (group 2). We surveyed the literature for age-specific incidence data about infections from both the groups. The age trends of the two groups show contrasting patterns. The incidence of infections by all group 1 pathogens showed a 2.28- to 28-fold increase in older ages. In group 2, 6 out of the 9 pathogens showed a significant declining trend in incidence with age. In both groups, there was greater mortality or morbidity among the infected in the old-age classes. These patterns are better explained by pathogen evolution than by age-related decline in immunity.


title: cis-acting sequences required for coronavirus infectious bronchitis virus defective-RNA replication and packaging.

text: The parts of the RNA genome of infectious bronchitis virus (IBV) required for replication and packaging of the RNA were investigated using deletion mutagenesis of a defective RNA (D-RNA) CD-61 (6.1 kb) containing a chloramphenicol acetyltransferase reporter gene. A D-RNA with the first 544, but not as few as 338, nucleotides (nt) of the 5' terminus was replicated; the 5' untranslated region (UTR) comprises 528 nt. Region I of the 3' UTR, adjacent to the nucleocapsid protein gene, comprised 212 nt and could be removed without impairment of replication or packaging of D-RNAs. A D-RNA with the final 338 nt, including the 293 nt in the highly conserved region II of the 3' UTR, was replicated. Thus, the 5'-terminal 544 nt and 3'-terminal 338 nt contained the necessary signals for RNA replication. Phylogenetic analysis of 19 strains of IBV and 3 strains of turkey coronavirus predicted a conserved stem-loop structure at the 5' end of region II of the 3' UTR. Removal of the predicted stem-loop structure abolished replication of the D-RNAs. D-RNAs in which replicase gene 1b-derived sequences had been removed or replaced with all the downstream genes were replicated well but were rescued poorly, suggesting inefficient packaging. However, no specific part of the 1b gene was required for efficient packaging.


docid: atqdpjun

title: Response of undergraduate medical students to SARS.


Identification of a noncanonical transcription initiation site for transcription of a subgenomic mRNA of mouse hepatitis virus.


title: SARS: how to manage future outbreaks?

text: Severe acute respiratory syndrome (SARS) was an unknown disease barely 3 years ago. After the World Health Organization declared the world SARS-free on 5 July 2003, there were episodic recurrences of SARS between September 2003 and May 2004, including 4 cases of laboratory-acquired SARS. SARS posed a mammoth challenge because of the impact of nosocomial transmission on healthcare manpower and facilities, and the resources needed for controlling and preventing further spread. Through worldwide scientific collaboration, the medical community has made much progress in unraveling its enigma, though much more needs to be discovered. This paper highlights how we can apply our knowledge of its epidemiology, mode of transmission, clinical course, ICU admission, complications, predictors of poor outcome, treatment and infection control to help us avert a catastrophic outbreak, and to manage our resources and patients. SARS preparedness and response planning must be flexible and dynamic so that appropriate measures can be implemented as an outbreak progresses. Even if SARS does not reemerge, the experience gained from such planning is valuable in preparing for threats of bioterrorism or a global avian influenza A (H5N1) virus pandemic.


title: Disaster and emergency management: Canadian nurses' perceptions of preparedness on hospital front lines.

text: INTRODUCTION Three years following the global outbreak of severe acute respiratory syndrome (SARS), a national, Web-based survey of Canadian nurses was conducted to assess perceptions of preparedness for disasters and access to support mechanisms, particularly for nurses in emergency and critical care units. HYPOTHESES The following hypotheses were tested: (1) nurses' sense of preparedness for infectious disease outbreaks and naturally occurring disasters will be higher than for chemical, biological, radiological, and nuclear (CBRN)-type disasters associated with terrorist attacks; (2) perceptions of preparedness will vary according to previous outbreak experience; and (3) perceptions of personal preparedness will be related to perceived institutional preparedness. METHODS Nurses from emergency departments and intensive care units across Canada were recruited via flyer mailouts and e-mail notices to complete a 30-minute online survey. RESULTS A total of 1,543 nurses completed the survey (90% female; 10% male). The results indicate that nurses feel unprepared to respond to large-scale disasters/attacks. The sense of preparedness varied according to the outbreak/disaster scenario with nurses feeling least prepared to respond to a CBRN event. A variety of socio-demographic factors, notably gender, previous outbreak experience (particularly with SARS), full-time vs. part-time job status, and region of employment also were related to perceptions of risk. Approximately 40% of respondents were unaware if their hospital had an emergency plan for a large-scale outbreak. Nurses reported inadequate access to resources to support disaster response capacity and expressed a low degree of confidence in the preparedness of Canadian healthcare institutions for future outbreaks. CONCLUSIONS Canadian nurses have indicated that considerably more training and information are needed to enhance preparedness for frontline healthcare workers as important members of the response community.


title: Caprine and bovine B rotaviruses in western France: group identification by Northern hybridization.

text: In a survey of coronavirus and rotavirus-induced neonatal diarrhoea in 373 calves from 284 farms, nine (2.4%) of the animals were found to be infected with non-group A rotaviruses when their faeces were analysed in a commercial ELISA and by PAGE. Seven out of eight 1-4- day-old kids from a single farm were also infected with similar viruses. In a mini-PAGE, all the viruses displayed group B- and/or E-like 4223 electrophoretypes. In Northern hybridizations with cDNA chemiluminescent probes specific to rotavirus groups A (RF strain), B (adult diarrhoea rotavirus, ADRV) and C (Cowden strain) all the viruses belonged to group B.


title: Human resources for health: overcoming the crisis.

text: In this analysis of the global workforce, the Joint Learning Initiative-a consortium of more than 100 health leaders-proposes that mobilisation and strengthening of human resources for health, neglected yet critical, is central to combating health crises in some of the world's poorest countries and for building sustainable health systems in all countries. Nearly all countries are challenged by worker shortage, skill mix imbalance, maldistribution, negative work environment, and weak knowledge base. Especially in the poorest countries, the workforce is under assault by HIV/AIDS, out-migration, and inadequate investment. Effective country strategies should be backed by international reinforcement. Ultimately, the crisis in human resources is a shared problem requiring shared responsibility for cooperative action. Alliances for action are recommended to strengthen the performance of all existing actors while expanding space and energy for fresh actors.


title: Mouse hepatitis virus type-2 infection in mice: an experimental model system of acute meningitis and hepatitis.

text: Infection with mouse hepatitis virus (MHV) strain A59 produces acute hepatitis, encephalitis, and chronic demyelination in mice. However, little is known about a closely related strain, MHV-2, which is only weakly neurotropic. To better understand the molecular basis of neurotropism of MHVs, we compared the pathogenesis and genomic sequence of MHV-2 with that of MHV-A59. Intracerebral injection of MHV-2 into 4-week-old C57B1/6 mice produces acute meningitis and hepatitis without encephalitis or chronic inflammatory demyelination. Sequence comparison between MHV-2 and MHV-A59 reveals 94-98% sequence identity of the replicase gene, 83-95% sequence identity of genes 2a, 3, 5b, 6, and 7, and marked difference in the sequence of genes, 2b, 4, and 5a. This information provides the basis for further studies exploring the mechanism of viral neurotropism and virus-induced demyelination.


title: Assessing the risk of observing multiple generations of Middle East respiratory syndrome (MERS) cases given an imported case.

text: To guide risk assessment, expected numbers of cases and generations were estimated, assuming a case importation of Middle East respiratory syndrome (MERS). Our analysis of 36 importation events yielded the risk of observing secondary transmission events at 22.7% (95% confidence interval: 19.3–25.1). The risks of observing generations 2, 3 and 4 were estimated at 10.5%, 6.1% and 3.9%, respectively. Countries at risk should be ready for highly variable outcomes following an importation of MERS.


title: Double Peripheral Venous and Arterial Cannulation for Extracorporeal Membrane Oxygenation in Combined Septic and Cardiogenic Shock.

text: BACKGROUND The use of venoarterial extracorporeal membrane oxygenation (va-ECMO) via peripheral cannulation for septic shock is limited by blood flow and increased afterload for the left ventricle. CASE REPORT A 15-year-old girl with acute myelogenous leukemia, suffering from severe septic and cardiogenic shock, was treated by venoarterial extracorporeal membrane oxygenation (va-ECMO). Sufficient extracorporeal blood flow matching the required oxygen demand could only be achieved by peripheral cannulation of both femoral arteries. Venous drainage was performed with a bicaval cannula inserted via the left V. femoralis. To accomplish left ventricular unloading, an additional drainage cannula was placed in the left atrium via percutaneous atrioseptostomy (va-va-ECMO). Cardiac function recovered and the girl was weaned from the ECMO on day 6. Successful allogenic stem cell transplantation took place 2 months later. CONCLUSIONS In patients with vasoplegic septic shock and impaired cardiac contractility, double peripheral venoarterial extracorporeal membrane oxygenation (va-va-ECMO) with transseptal left atrial venting can by a lifesaving option.


title: Initial otolaryngological manifestations of severe acute respiratory syndrome in Taiwan.

text: OBJECTIVE To provide a clue for screening severe acute respiratory syndrome (SARS), a highly transmissible disease with health care workers at particular risk, in the early stage from an otolaryngological perspective. DESIGN Prospective study. SETTING Community hospital. PATIENTS Between April 17 and April 26, 2003, 32 consecutive patients with SARS were encountered. Investigation consisted of local examination of ear, nose, and throat fields; palpation of the nuchal areas; and plain chest radiography. Analyses of throat swab samples using reverse transcription-polymerase chain reaction were conducted. RESULTS Clinical manifestations included fever in 31 patients (97%), followed by cough, dyspnea, chill, headache, sore throat, diarrhea, rhinorrhea, and otalgia. Neither the pharyngeal wall nor the tonsillar area demonstrated hyperemia. There was no lymphadenopathy in the neck. Plain chest radiographs revealed consolidation in 25 (78%) of 32 patients. Results of reverse transcription-polymerase chain reaction analysis targeting the novel coronavirus present in throat swab samples were positive in 19 (66%) of 29 patients tested. Twenty-eight patients required supplemental oxygen, and 14 patients were intubated with mechanical ventilation. Twenty-eight patients survived and 4 patients died. CONCLUSIONS When presented with a patient with flulike symptoms such as fever and/or cough, but no pharyngeal hyperemia or neck lymphadenopathy, physicians should be alerted to the possibility of SARS. In contrast, evidence of inflammatory signs in the otolaryngological field may explain the flulike symptoms, and serve as a differential diagnostic tool between influenza and SARS.


Excellence in research and education at the John A. Burns School of Medicine: a tribute to Edwin Cadman's vision.


Treatment of staphylococcal infection.


title: Emerging infectious diseases not covered by routine vaccination in Europe in 2010-2015--the review of WHO and ECDC notifications for the National IHR Focal Point in Poland.

text: The National IHR Focal Point is a center set up by each Member State of the World Health Organization (WHO) in accordance with the International Health Regulations (2005). The International Health Regulations (IHR) were adopted on 23 May 2005 at the World Health Assembly and entered into force since 15 June 2007 as the legal instrument designed to help protect all countries from uncontrolled international spread of diseases and other urgent public health threats. According to Article 2 of IHR the purpose and scope of these Regulations are to prevent, protect against, control and provide a public health response to the international spread of disease in ways that are commensurate with and restricted to public health risks, and which avoid unnecessary interference with international traffic and trade. Primarily, the scope of IHR is to establish a system of early warning (in accordance with Article 6 and 7) with the functioning in each country National IHR Focal Point which is available at any time to communicate with WHO IHR Contact Points and other entities. The tasks of the National IHR Fo- cal Point in Poland which was appointed by the Minister of Health and runs in the Department of Epidemiology, National Institute of Public Health--National Institute of Hygiene from 1 September 2007 are the notification of events that may constitute a public health emergency of international concern occurring in Poland or abroad and the dissemination of this information to the WHO, other National IHR Focal Points or competent authorities responsible for public health. The task of the National IHR Focal Point in Poland is also the dissemination of WHO and ECDC notifications, including recommendation and risk assessment documents. The aim of this work is the review of WHO and ECDC notifications received by National IHR Focal Point in Poland in the period from 2010 to 2015 which were related to emerging infectious diseases not covered by routine vaccination programs or for which there are no effective vaccines that have occurred in the WHO European Region. The review includes verotoxin-producing Escherichia coli O104: H4 infections, MERS-CoV infections, Ebola virus disease, malaria, dengue fever, West Nile fever, chikungunya and cholera.


title: Coronavirus particle assembly: primary structure requirements of the membrane protein.

text: Coronavirus-like particles morphologically similar to normal virions are assembled when genes encoding the viral membrane proteins M and E are coexpressed in eukaryotic cells. Using this envelope assembly assay, we have studied the primary sequence requirements for particle formation of the mouse hepatitis virus (MHV) M protein, the major protein of the coronavirion membrane. Our results show that each of the different domains of the protein is important. Mutations (deletions, insertions, point mutations) in the luminal domain, the transmembrane domains, the amphiphilic domain, or the carboxy-terminal domain had effects on the assembly of M into enveloped particles. Strikingly, the extreme carboxy-terminal residue is crucial. Deletion of this single residue abolished particle assembly almost completely; most substitutions were strongly inhibitory. Site-directed mutations in the carboxy terminus of M were also incorporated into the MHV genome by targeted recombination. The results supported a critical role for this domain of M in viral assembly, although the M carboxy terminus was more tolerant of alteration in the complete virion than in virus-like particles, likely because of the stabilization of virions by additional intermolecular interactions. Interestingly, glycosylation of M appeared not essential for assembly. Mutations in the luminal domain that abolished the normal O glycosylation of the protein or created an N-glycosylated form had no effect. Mutant M proteins unable to form virus-like particles were found to inhibit the budding of assembly-competent M in a concentration-dependent manner. However, assembly-competent M was able to rescue assembly-incompetent M when the latter was present in low amounts. These observations support the existence of interactions between M molecules that are thought to be the driving force in coronavirus envelope assembly.


title: A cluster-randomized controlled trial evaluating the effect of a handwashing-promotion program in Chinese primary schools.

text: Intensive handwashing promotion can reduce diarrheal and respiratory disease incidence. To determine whether less intensive, more scalable interventions can improve health, we evaluated a school-based handwashing program. We randomized 87 Chinese schools to usual practices: standard intervention (handwashing program) or expanded intervention (handwashing program, soap for school sinks, and peer hygiene monitors). We compared student absence rates, adjusting for cluster design. In control schools, children experienced a median 2.0 episodes (median 2.6 days) of absence per 100 student-weeks. In standard intervention schools, there were a median 1.2 episodes (P = 0.08) and 1.9 days (P = 0.14) of absence per 100 student-weeks. Children in expanded intervention schools experienced a median 1.2 episodes (P = 0.03) and 1.2 days (P = 0.03) of absence per 100 student-weeks. Provision of a large-scale handwashing promotion program and soap was associated with significantly reduced absenteeism. Similar programs could improve the health of children worldwide.


title: A self-tuning method for one-chip SNP identification.

text: Current methods for interpreting oligonucleotide-based SNP-detection microarrays, SNP chips, are based on statistics and require extensive parameter tuning as well as extremely high-resolution images of the chip being processed. We present a method, based on a simple data-classification technique called nearest-neighbors that, on haploid organisms, produces results comparable to the published results of the leading statistical methods and requires very little in the way of parameter tuning. Furthermore, it can interpret SNP chips using lower-resolution scanners of the type more typically used in current microarray experiments. Along with our algorithm, we present the results of a SNP-detection experiment where, when independently applying this algorithm to six identical SARS SNP chips, we correctly identify all 24 SNPs in a particular strain of the SARS virus, with between 6 and 13 false positives across the six experiments.


Sequences involved in the replication of coronaviruses.


title: Long-term effects of azathioprine in rheumatoid arthritis.

text: Efficacy and safety of azathioprine in 'high' and 'low' dose regimens in rheumatoid arthritis (RA), both in short-term studies and in follow-up over 40 months, have previously been shown. In the present report, 36 patients with RA treated with azathioprine (group I) and 49 age-matched patients with RA (group II), were studied to detect potential early markers of malignancy. Chest x-rays were similar to both groups. One patient in group I had a positive PAP smear and was subsequently found to have uterine carcinoma. Alpha-fetoprotein was positive in one patient in group I and none in group II. CEA was negative in all patients in group I, but positive in seven in group II. On chromosomal analysis group I showed a greater frequency of breakage. Group I showed lower serum folates and a highly significant number of megaloblastic features in marrow aspirates. In group I seven tumours, three being malignant, occurred while taking azathioprine, and in group II six tumours, one malignant, were identified (p = 0.17). The apparent increased risk of malignancy previously suggested by others warrants further studies with larger populations and over a continuous longer period.


title: The Duality of Fgl2 - Secreted Immune Checkpoint Regulator Versus Membrane-Associated Procoagulant: Therapeutic Potential and Implications.

text: Fibrinogen-like protein 2 (Fgl2), a member of the fibrinogen family, can be expressed as a membrane-associated protein with coagulation activity or in a secreted form possessing unique immune suppressive functions. The biological importance of Fgl2 is evident within viral-induced fibrin depositing inflammatory diseases and malignancies and provides a compelling rationale for Fgl2 expression to not only be considered as a disease biomarker but also as a therapeutic target. This article will provide a comprehensive review of the currently known biological properties of Fgl2 and clarifies future scientific directives.


docid: dmdnbktk

title: Attaching and effacing lesions in the intestines of two calves associated with natural infection with Escherichia coli O26:H11.


title: Survey of dogs in Japan for group 2 canine coronavirus infection.

text: Specimens obtained from 96 dogs with respiratory and enteric clinical signs in Japan were retrospectively examined for group 2 coronavirus by reverse transcription-PCR. Two dogs were found to be positive. Phylogenetic analysis of the spike gene indicated that they were most probably related to the canine respiratory coronavirus recently described in the United Kingdom.


title: Job stress, burnout, depression symptoms, and physical health among Chinese university teachers.

text: The aim of the present study was to investigate the relationships among job stress, burnout, depression, and health among university teachers in China. Using a stratified random sampling method, a sample of 300 university teachers completed the Occupational Stress Indicator-2 (OSI-2), Maslach Burnout Inventory-General Survey (MBI-GS), Beck Depression Inventory (BDI), and 36-item Short Form Health Survey (SF-36). Path analysis showed that burnout was a mediator among job stress, the occurrence and exacerbation of depressive symptoms, and poor physical health.


title: Manganese-dependent polioviruses caused by mutations within the viral polymerase.

text: Viral RNA-dependent RNA polymerases exhibit great sequence diversity. Only six core amino acids are conserved across all polymerases of positive-strand RNA viruses of eukaryotes. While exploring the function of one of these completely conserved residues, asparagine 297 in the prototypic poliovirus polymerase 3D(pol), we identified three viable mutants with noncanonical amino acids at this conserved position. Although asparagine 297 could be replaced by glycine or alanine in these mutants, the viruses exhibited Mn(2+)-dependent RNA replication and viral growth. All known RNA polymerases and replicative polymerases of bacterial, eukaryotic, and viral organisms are thought to be magnesium dependent in vivo, and therefore these mutant polioviruses may represent the first viruses with a requirement for an alternative polymerase cation. These results demonstrate the extreme functional flexibility of viral RNA-dependent RNA polymerases. Furthermore, the finding that strictly conserved residues in the nucleotide binding pocket of the polymerase can be altered in a manner that supports virus production suggests that drugs targeting this region of the enzyme will still be susceptible to the problem of drug-resistant escape mutants.


title: Coronavirus IBV: removal of spike glycopolypeptide S1 by urea abolishes infectivity and haemagglutination but not attachment to cells.

text: Urea has been used to remove the S1 spike glycopolypeptide from avian infectious bronchitis virus (IBV) strains M41 and Beaudette, without removing the S2 spike-anchoring glycopolypeptide. Reduction of the pH to 2.9 did not cause release of S1 although some S1 was released spontaneously from IBV Beaudette at pH 7.4. Virus that lacked S1 was no longer infectious or able to cause haemagglutination (HA). However, radiolabelled IBV that lacked S1 attached to erythrocytes and chick kidney cells to the same or similar extent as did intact virus. Treatment of IBV with a phospholipase C preparation, required to make IBV cause HA, did not increase binding of IBV to erythrocytes. The results indicate that while the attachment to cells of virus that lacks S1 is qualitatively different from that of intact virus, the decline in infectivity is the consequence of the loss of some other spike function.


title: Temperature-sensitive acetylesterase activity of haemagglutinin-esterase specified by respiratory bovine coronaviruses.

text: Numerous respiratory bovine coronaviruses (RBCV) were isolated recently from nasal swab samples and lung tissues of feedlot cattle with acute respiratory tract disease. These newly emerging RBCV isolates exhibited distinct phenotypic features that differentiated them from enteropathogenic bovine coronaviruses (EBCV). The RBCV strains had a receptor-destroying enzyme function mediated by acetylesterase (AE) activity of the haemagglutinin-esterase (HE) glycoprotein. The HE genes of wild-type EBCV strain LY138 and RBCV strains OK-0514 (OK) and LSU-94LSS-051 (LSU) were cloned, sequenced and transiently expressed in COS-7 cells. The enzymic properties of HE proteins in COS-7 cellular extracts and in purified virus preparations were assayed at room temperature, 37 degrees C and 39 degrees C by two different assays. One assay used p-nitrophenyl acetate (PNPA) as substrate and detected serine-esterase activity; the second assay monitored AE function with bovine submaxillary mucin (BSM) as substrate. The PNPA tests confirmed that HE proteins of EBCV and RBCV were functionally expressed in transfected COS-7 cells. Time-dependent determination of the AE activity of purified RBCV OK and LSU particles showed lower AE activity at 39 degrees C than at 37 degrees C, whereas the purified EBCV LY particles retained full AE activity at both 37 degrees C and 39 degrees C. Transiently expressed RBCV HE exhibited a marked reduction of AE activity after 40 min of assay time at 37 degrees C. In contrast, the AE activity of the transiently expressed EBCV HE remained stable beyond 40 min. The deduced amino-acid sequences of the HE proteins specified by the RBCV strains OK and LSU contained specific amino-acid changes in comparison with the EBCV LY strain, which may be responsible for the observed enzymic differences. These results are consistent with the hypothesis that RBCV strains have evolved to selectivelyreplicate in respiratory tissues and that HE may play a role in this tissue tropism.


Using Technology in Biosurveillance and Epidemic Management.


title: Periodic monitoring of commercial turkeys for enteric viruses indicates continuous presence of astrovirus and rotavirus on the farms.

text: A longitudinal survey to detect enteric viruses in intestinal contents collected from turkeys in eight commercial operations and one research facility was performed using molecular detection methods. Intestinal contents were collected from turkeys prior to placement, with each flock resampled at 2, 4, 6, 8, 10, and 12 wk of age. The samples were screened for astrovirus, rotavirus, reovirus, and turkey coronavirus (TCoV) by a reverse transcriptase and polymerase chain reaction (RT-PCR), and for groups 1 and 2 adenovirus by PCR. Rotavirus was the only virus detected prior to placement (7 of 16 samples examined). All of the commercial flocks were positive for rotavirus and astrovirus from 2 until 6 wk of age, and most were intermittently positive until 12 wk of age, when the birds were processed. Of the 96 samples collected from birds on the farms, 89.5% were positive for astrovirus, and 67.7% were positive for rotavirus. All flocks were negative for TCoV, reovirus, and group 1 adenovirus at all time points, and positive for group 2 adenovirus (hemorrhagic enteritis virus) at 6 wk of age. All the flocks monitored were considered healthy or normal by field personnel. Turkeys placed on research facilities that had been empty for months and thoroughly cleaned had higher body weights and lower feed conversion rates at 5 wk of age when compared to turkeys placed on commercial farms. Intestinal samples collected at 1, 2, and 3 wk of age from these turkeys were free of enteric viruses. This report demonstrates that astroviruses and rotaviruses may be present within a turkey flock through the life of the flock. Comparison of infected birds with one group of turkeys that were negative for enteric viruses by the methods used here suggests that astrovirus and/or rotavirus may affect production. The full impact on flock performance needs to be further determined.


title: Isolation of porcine respiratory coronavirus from pigs affected with porcine reproductive and respiratory syndrome.

text: Four cytopathogenic viruses were isolated in CPK cells derived from porcine kidneys from tonsils and lungs of 3 of 15 pigs affected with porcine reproductive and respiratory syndrome virus. Physicochemically and morphologically, the isolates were similar to a coronavirus. The isolates were not distinguished from transmissible gastroenteritis virus (TGEV) by a neutralization test using polyclonal antibodies, but differentiated from TGEV by monoclonal antibodies capable of discriminating between TGEV and porcine respiratory coronavirus (PRCV), indicating that the isolates were PRCV. In a serological survey of 30 serum samples each collected from about 50 days old pigs in the 2 affected farms, 29 (97%) and 15 (50%) sera were positive for neutralizing antibody against the isolate with the titers ranging from 2 to 64, respectively.


title: Mice immunogenicity after vaccination by DNA vaccines containing individual genes of a new type of reovirus.

text: In this study, we investigated humoral and cellular immune responses in mice to DNA vaccines containing individual S or M genes of a new type of reovirus (nRV) isolate from a severe acute respiratory syndrome (SARS) patient in Beijing, China. Mice were immunized intramuscularly (i.m.) with 100 μg of S1, S2, S3, S4, M1, M2, and M3 DNA vaccine each 4 times in 2-week intervals and assayed for humoral IgG, IgG1, IgG2, and IgG2b antibodies by ELISA and for cellular immune response, particularly IFN-γ induction by ELISpot assay. Moreover, CD4+ and CD8+ T cell levels in peripheral blood mononuclear cells were assayed by flow cytometry. We found that all DNA vaccines induced IgG antibodies, predominantly of the IgG2a class and S3 DNA vaccine was the strongest inducer. M2 and S3 DNA vaccines elicited Th1- and Th2-based immune responses, respectively, while S1 and M3 DNA vaccines induced a mixed Th1/Th2 response. M1, S2, and S4 DNA vaccines were poorly immunogenic. To our knowledge, this is the first report characterizing mammalian reovirus DNA vaccines applied to a mouse model.


title: Validation of Impact of Events Scale in nurses under threat of contagion by severe acute respiratory syndrome.

text: The purpose of the present study was to validate the Impact of Events Scale (IES) in a sample of nurses working under threat of severe acute respiratory syndrome (SARS). The internal consistency, construct validity and convergent validity of the instrument were examined in a sample of 128 nurses during the SARS outbreak in Taiwan. Principal component analysis followed by a quartimax rotation were used to derive a two-factor solution, labeled intrusion (factor 1) and avoidance (factor 2), with both factors accounting for 50.7% of the explained variance. The total Cronbach's alpha of 0.90 reflected the good internal consistency of the instrument. Correlation with the Symptoms Checklist 90-R demonstrated the convergent validity of the IES. In conclusion, the IES can be used as a convenient, reliable and valid instrument for evaluation of the psychological distress of nurses working with the highly contagious disease.


docid: mpvwovqg

title: Post-thyroidectomy hypoparathyroidism, what should we keep in mind?

text: AIM Hypoparathyroidism and the resulting hypocalcemia is a common iatrogenic complication following surgical procedures to the neck, and commonly, to the thyroid gland. The aim of this study was to review the available literature to summarize current data related to the development of hypoparathyroidism after thyroid surgery. MATERIALS AND METHODS An analysis of the surgical literature was performed using the search engine EMBASE and PubMed with particular reference to the principal risk factors related to the post-thyroid surgery hypoparathyroidism. Moreover the medical records of 345 patients, who underwent thyroid surgery at the Section of Endocrine Surgery - Department of Medical and Surgical Sciences, Advanced Technologies "G. Ingrassia" University of Catania, Italy, were also examined. RESULTS The definition of hypoparathyroidism varies widely in literature. There is a wide range of reported incidences of transient and permanent postoperative hypoparathyroidism in literature. Extensive surgery, malignant disease and concomitant central and/or lateral neck dissection, autoimmune or Grave's' disease and re-operation represent the most recognized risk factors. A wide consensus exists about transplanting a parathyroid gland when it seems to be nonviable. DISCUSSION Although many improvements have been done in surgical technique, hypoparathyroidism seems to be one of the most common complication after total thyroidectomy. In this study we discuss the diagnosis methods and the importance of its early prediction regardless it is transient or permanent. Recovery time is under discussion: some author consider a gland permanent injury if restoring functionality does not occur within 6 months other 12 months. To assess the parathyroid gland function some authors have been using postoperative PTH level as a useful tool for predicting hypocalcemia. CONCLUSIONS Many efforts are required to address the problem of a consensus on best define these complications. An early low PTH level after total thyroidectomy is associated with a high risk of permanent hypoparathyroidism and normal levels usually exclude long-term parathyroid glands dysfunction. However, the cost of the PTH assay may limit its widespread application. Despite the reassuring presence of new therapeutic strategies, intra-operative preservation of the parathyroid glands is the best prophylaxis to avoid postoperative hypocalcemia after total thyroidectomy. KEY WORDS Hypoparathyroidism, Thyroid surgery.


docid: qveigqby

title: J-Pouch Prolapse as a Late Complication of Restorative Proctocolectomy.


title: A novel method to prevent secondary exposure of medical and rescue personnel to toxic materials under biochemical hazard conditions using microwave radar and infrared thermography.

text: In order to prevent secondary exposure of medical personnel to toxic materials under biochemical hazard conditions, we performed a noncontact determination of exposure to toxic conditions via 1215-MHz microwave radar and thermography. A toxic condition was induced by intravenous administration of lipopolysaccharide (LPS) in rabbits. The exposure to LPS was determined by linear discriminant analysis using non-contact derived variables.


title: Induction of specific immune responses by severe acute respiratory syndrome coronavirus spike DNA vaccine with or without interleukin-2 immunization using different vaccination routes in mice.

text: DNA vaccines induce humoral and cellular immune responses in animal models and humans. To analyze the immunogenicity of the severe acute respiratory syndrome (SARS) coronavirus (CoV), SARS-CoV, spike DNA vaccine and the immunoregulatory activity of interleukin-2 (IL-2), DNA vaccine plasmids pcDNA-S and pcDNA-IL-2 were constructed and inoculated into BALB/c mice with or without pcDNA-IL-2 by using three different immunization routes (the intramuscular route, electroporation, or the oral route with live attenuated Salmonella enterica serovar Typhimurium). The cellular and humoral immune responses were assessed by enzyme-linked immunosorbent assays, lymphocyte proliferation assays, enzyme-linked immunospot assays, and fluorescence-activated cell sorter analyses. The results showed that specific humoral and cellular immunities could be induced in mice by inoculating them with SARS-CoV spike DNA vaccine alone or by coinoculation with IL-2-expressing plasmids. In addition, the immune response levels in the coinoculation groups were significantly higher than those in groups receiving the spike DNA vaccine alone. The comparison between the three vaccination routes indicated that oral vaccination evoked a vigorous T-cell response and a weak response predominantly with subclass immunoglobulin G2a (IgG2a) antibody. However, intramuscular immunization evoked a vigorous antibody response and a weak T-cell response, and vaccination by electroporation evoked a vigorous response with a predominant subclass IgG1 antibody response and a moderate T-cell response. Our findings show that the spike DNA vaccine has good immunogenicity and can induce specific humoral and cellular immunities in BALB/c mice, while IL-2 plays an immunoadjuvant role and enhances the humoral and cellular immune responses. Different vaccination routes also evoke distinct immune responses. This study provides basic information for the design of DNA vaccines against SARS-CoV.


docid: npjnnkid

title: Preparation of Protein-containing Extracts from Microbiota-rich Intestinal Contents.

text: The contribution of microbiota in regulating multiple physiological and pathological host responses has been studied intensively in recent years. Evidence suggests that commensal microbiota can directly modulate different populations of cells of the immune system (e.g., Ivanov et al., 2008; Atarashi et al., 2011). Recently, we showed that protein extracts from gut commensal microbiota can activate retina-specific T cells, allowing these autoreactive T cells to then break through the blood-retinal barrier and trigger autoimmune uveitis in the recipient (Horai et al., 2015). The protocol below describes the method to prepare intestinal protein-rich extracts that can be used in various in vitro andin vivo immunological studies.


title: Presumptive abortive human rabies - Texas, 2009.

text: Rabies is a serious zoonotic disease. Recovery has been well documented in only six human patients worldwide. Five of those patients had received rabies vaccinations before illness; one had not received rabies vaccination but survived infection after prolonged intensive care. In most of these survivors, moderate to profound neurologic sequelae occurred. In all six survivors, rabies was diagnosed based on exposure history, compatible clinical symptoms, and detection of rabies virus-neutralizing antibodies (VNA). This report describes the clinical course and laboratory findings of an adolescent girl with encephalitis who had not had rabies vaccination and who had been exposed to bats 2 months before illness. Antibodies to rabies virus were detected in specimens of the girl's serum and cerebrospinal fluid (CSF) by indirect fluorescent antibody test (IFA). However, the presence of rabies VNA was not detected until after she had received single doses of rabies vaccine and human rabies immune globulin (HRIG). Although the patient required multiple hospitalizations and follow-up visits for recurrent neurologic symptoms, she survived without intensive care. No alternate etiology was determined, and abortive human rabies (defined in this report as recovery from rabies without intensive care) was diagnosed. Public education should emphasize avoiding exposure to bats and other potentially rabid wildlife and seeking prompt medical attention after exposure to such animals. Rabies is preventable if rabies immune globulin and vaccine are administered soon after an exposure; however, this case also suggests the rare possibility that abortive rabies can occur in humans and might go unrecognized.


title: Lack of SARS transmission among healthcare workers, United States.

text: Healthcare workers accounted for a large proportion of persons with severe acute respiratory syndrome (SARS) during the worldwide epidemic of early 2003. We conducted an investigation of healthcare workers exposed to laboratory-confirmed SARS patients in the United States to evaluate infection-control practices and possible SARS-associated coronavirus (SARS-CoV) transmission. We identified 110 healthcare workers with exposure within droplet range (i.e., 3 feet) to six SARS-CoV-positive patients. Forty-five healthcare workers had exposure without any mask use, 72 had exposure without eye protection, and 40 reported direct skin-to-skin contact. Potential droplet- and aerosol-generating procedures were infrequent: 5% of healthcare workers manipulated a patient's airway, and 4% administered aerosolized medication. Despite numerous unprotected exposures, there was no serologic evidence of healthcare-related SARS-CoV transmission. Lack of transmission in the United States may be related to the relative absence of high-risk procedures or patients, factors that may place healthcare workers at higher risk for infection.


In Defense of SAPPHIRE.


title: Feline leukemia virus in a captive bobcat.

text: An 11-mo-old captive-bred male neutered bobcat (Felis rufus) presented with lethargy, anorexia, leukopenia, neutropenia, lymphopenia, and nonregenerative anemia. The animal was diagnosed as feline leukemia virus (FeLV) positive by immunofluorescent antibody and enzyme-linked immunosorbant assay (ELISA) testing. It died despite supportive care. Pathologic examination revealed multifocal non-suppurative encephalitis, diffuse interstitial pneumonia, multifocal hepatocellular necrosis, non-suppurative peritonitis, and lymphoid depletion. FeLV was isolated from peripheral blood mononuclear cells, bone marrow, spleen, and lymph node. FeLV-specific gag sequences were amplified by DNA polymerase chain reaction (PCR) and aligned with known domestic cat FeLV's. The source of the virus was speculated to be a domestic cat that was a surrogate nurse. Case reports of FeLV in nondomestic felids are few, and FeLV does not appear to be enzootic in wild felids, except European wildcats (Felis silvestris) in France and Scotland. Introduction of FeLV into free-living and captive nondomestic felid populations could have serious consequences for their health and survival. Measures to prevent the introduction of this virus to nondomestic felids are warranted.


docid: qfcesatx

title: Radiofrequency ablation for the management of liver tumors.

text: BACKGROUND Radiofrequency ablation has recently become a viable treatment option for unresectable primary or secondary lesions confined to the liver. OBJECTIVE To study the local therapeutic efficacy, side effects and complications of radiofrequency ablation for the treatment of hepatocellular carcinoma and liver metastases. This is the first reported experience of radiofrequency ablation for treating malignant hepatic tumors in Israel. METHODS Fifteen consecutive patients, aged 53-73 years, with 23 lesions (8 patients with HCC and 7 with secondary liver tumors) underwent radiofrequency ablation under general anesthesia. RITA nine-array 5 cm thermal ablation catheter and the model 1500 generator were used. The mean diameter of all tumors was 4.28 cm (range 1-10 cm). Three lesions were 1-3 cm in diameter (small), 17 lesions measured 3.1-5 cm (medium), and 3 measured 5.1-10 cm (large). RESULTS Complete necrosis was found in 8 (66%) of 12 HCCs by computed tomography scan. Of the remainder, diffuse tumor recurrence was demonstrated in three lesions (25%) after lipiodol injection and there was one local tumor recurrence. In the metastases group complete necrosis was found in 5 of 11 lesions (45%). One major complication (peritonitis) was treated with antibiotics and four (26%) minor complications (right pleural effusion, small subcapsular hematoma) were monitored. CONCLUSIONS Radiofrequency ablation appears to be an effective, safe and relatively simple procedure for the treatment of liver tumors.


SARS coronavirus without reservoir originated from an unnatural evolution, experienced the reverse evolution, and finally disappeared in the world.


title: Development and application of an enzyme immunoassay for coronavirus OC43 antibody in acute respiratory illness.

text: Study of coronavirus OC43 infections has been limited because of the lack of sensitive cell culture systems and serologic assays. To improve this circumstance, we developed an indirect enzyme immunoassay (EIA) to detect serum antibody to OC43. Antigen (100 ng) prepared by polyethylene glycol precipitation provided optimal results without a postcoat procedure. Evaluation of intraplate variation indicated that a > or = 2.5-fold increase in serum titer was significant. Sixteen of 18 (89%) paired serum samples with previously identified, reproducible increases in the level of hemagglutination inhibition (HAI) antibody to OC43 also showed significant increases as detected by EIA. Specificity for the EIA was established with paired sera obtained from persons given influenza immunizations or experiencing a respiratory infection. No rise in antibody titers occurred among 33 persons with documented coronavirus 229E infection. EIA was then performed on each of 419 paired serum samples from ambulatory chronic obstructive pulmonary disease patients and healthy older adults, from asthmatic adults presenting for emergency room treatment, and from persons hospitalized with acute respiratory symptoms. Twenty-three antibody rises to OC43 were detected; only nine of these were detected by the HAI test, and the HAI test did not detect any increases in antibody titers that were not detected by EIA. Nineteen of 25 coronavirus OC43 infections for which a month of infection could be assigned occurred between November and February. Overall, 4.4% of acute respiratory illnesses in the studied populations were associated with a coronavirus OC43 infection.


title: Developments in antiviral drug design, discovery and development in 2004.

text: This article summarizes key aspects of progress made during 2004 toward the design, discovery and development of antiviral agents for clinical use. Important developments in the identification, characterization and clinical utility of inhibitors of human immunodeficiency virus; the hepatitis viruses, hepatitis B, hepatitis C; the herpes family of viruses, herpes simplex viruses 1 and 2, varicella zoster virus, Epstein-Barr virus and human cytomegalovirus; the respiratory viruses, influenza, respiratory syncytial virus, human metapneumovirus, picornaviruses, measles and the severe acute respiratory syndrome coronavirus; human papilloma virus; rotavirus; Ebola virus and West Nile virus, are reviewed.


title: Structure-based preliminary analysis of immunity and virulence of SARS coronavirus.

text: The research on SARS-associated coronavirus (SARS-CoV) has not stopped since its discovery, but the pathogenesis of SARS is still unclear. To explore the possible molecular mechanisms of the invasion and virulence of SARS-CoV, we investigated the structural basis of the viral proteins using computational biology. Forty-five motifs relating to superantigens, toxins and other bioactive molecules were detected in the proteins of SARS-CoV. The results showed that the distribution of the motifs varied in different proteins. Enzyme-like motifs were located in the R protein, while ICAM- 1-like and toxin-like molecules were located in the spike, envelop, nucleocapsid, PUP1, PUP 2 and PUP 4 proteins. Comparison of SARS-CoV with other viruses (OC43, PEDV, HRSV, HHerpV and HAdenoV) showed that each group of motifs was different for each type of virus. Data suggest that the proteins of SARS-CoV with toxic motifs might play crucial roles in targeting host cells and interfering with the immune system. This study provides new information for drug and vaccine design, as well as therapeutic strategies against SARS.


title: Comparison of virus isolation, immunohistochemistry, and reverse transcriptase-polymerase chain reaction procedures for detection of turkey coronavirus.

text: A reverse transcriptase-polymerase chain reaction (RT-PCR) procedure and two monoclonal antibody (MAb)-based immunohistochemical procedures were developed for detection of turkey coronavirus (TCV) in tissues and intestinal contents/dropping samples. The RT-PCR, MAb-based fluorescent antibody (FA), and MAb-based immunoperoxidase (IP) procedures were compared with virus isolation (VI) for detection of TCV in experimentally infected turkeys. TCV was detected in experimentally infected turkeys as early as day 1 postexposure (PE) by each of the four detection procedures. TCV was detected as late as day 35 PE by FA or IP and days 42 and 49 PE by VI and RT-PCR, respectively. With VI as a reference, sensitivity and specificity of RT-PCR were 93% and 92%, respectively; specificity of both FA and IP was 96%, and sensitivities were 69% and 61%, respectively. Each of the examined procedures was highly specific, but the RT-PCR procedure was also highly sensitive. These findings demonstrate the utility of both immunohistochemistry and RT-PCR for detection of TCV. In addition, the findings indicate that RT-PCR is a highly sensitive and specific alternative to conventional diagnostic procedures.


title: Single-step multiplex conventional and real-time reverse transcription polymerase chain reaction assays for simultaneous detection and subtype differentiation of Influenza A virus in swine.

text: Because pigs are considered intermediate hosts for the emergence of novel influenza virus reassortants with associated zoonotic potential, monitoring and characterization of circulating influenza viruses in pigs are important for adequate control of infection. For this, rapid molecular diagnostic methods other than immunoassays are needed. Three novel single-step multiplex reverse transcription polymerase chain reaction (RT-PCR) assays were developed in the current study for simultaneous detection and subtype differentiation of Influenza A virus in pigs. A conventional single-step pentaplex RT-PCR was designed for concomitant detection of the generic matrix (M) gene, hemagglutinin H1 and H3, and neuraminidase N1 and N2 genes of Swine influenza virus (SIV). In the other 2 single-step tetraplex real-time RT-PCR assays, the primers and fluorescent probes were targeted for the simultaneous detection of common M, H1, H3, and N2 SIV genes (first assay), and for M, H1, and H3 SIV genes and the H5 gene of highly pathogenic avian influenza virus of Eurasian lineage (second assay). The real-time RT-PCR assays had detection sensitivity limits ranging from 10(1) to 10(3) copies of respective in vitro RNA transcripts of M, H1, H3, H5, and N2 genes. The multiplex assays were evaluated by using SIV isolates, clinical specimens, and the appropriate synthetic template. The recent H1N1 pandemic strain isolated from pigs also was tested in simplex RT-PCR and real-time RT-PCR assays with the H1 primers and probes. The efficacy of the multiplex RT-PCR and real-time RT-PCR shows the suitability of multiplex RT-PCR and real-time RT-PCR for rapid subtype identification and monitoring in North American pigs of Influenza A virus.


title: Brazilian recommendations of mechanical ventilation 2013. Part I.

text: Perspectives on invasive and noninvasive ventilatory support for critically ill patients are evolving, as much evidence indicates that ventilation may have positive effects on patient survival and the quality of the care provided in intensive care units in Brazil. For those reasons, the Brazilian Association of Intensive Care Medicine (Associação de Medicina Intensiva Brasileira - AMIB) and the Brazilian Thoracic Society (Sociedade Brasileira de Pneumonia e Tisiologia - SBPT), represented by the Mechanical Ventilation Committee and the Commission of Intensive Therapy, respectively, decided to review the literature and draft recommendations for mechanical ventilation with the goal of creating a document for bedside guidance as to the best practices on mechanical ventilation available to their members. The document was based on the available evidence regarding 29 subtopics selected as the most relevant for the subject of interest. The project was developed in several stages, during which the selected topics were distributed among experts recommended by both societies with recent publications on the subject of interest and/or significant teaching and research activity in the field of mechanical ventilation in Brazil. The experts were divided into pairs that were charged with performing a thorough review of the international literature on each topic. All the experts met at the Forum on Mechanical Ventilation, which was held at the headquarters of AMIB in São Paulo on August 3 and 4, 2013, to collaboratively draft the final text corresponding to each sub-topic, which was presented to, appraised, discussed and approved in a plenary session that included all 58 participants and aimed to create the final document.


title: Adaptation of dogs to the amputation of a limb and their owners' satisfaction with the procedure.

text: Forty-four Dutch dog owners were interviewed by telephone about their experiences with their dog after it had had a limb amputated. Most of the dogs adapted to walking on three legs within a month, sooner than most of the owners had expected. There were few complications after the amputation, although changes in behaviour were observed in approximately one-third of the dogs.


docid: bnbbwhrk

title: Assembly of spikes into coronavirus particles is mediated by the carboxy-terminal domain of the spike protein.

text: The type I glycoprotein S of coronavirus, trimers of which constitute the typical viral spikes, is assembled into virions through noncovalent interactions with the M protein. Here we demonstrate that incorporation is mediated by the short carboxy-terminal segment comprising the transmembrane and endodomain. To this aim, we used the virus-like particle (VLP) system that we developed earlier for the mouse hepatitis virus strain A59 (MHV-A59) and which we describe now also for the unrelated coronavirus feline infectious peritonitis virus (FIPV; strain 79-1146). Two chimeric MHV-FIPV S proteins were constructed, consisting of the ectodomain of the one virus and the transmembrane and endodomain of the other. These proteins were tested for their incorporation into VLPs of either species. They were found to assemble only into viral particles of the species from which their carboxy-terminal domain originated. Thus, the 64-terminal-residue sequence suffices to draw the 1308 (MHV)- or 1433 (FIPV)-amino-acid-long mature S protein into VLPs. Both chimeric S proteins appeared to cause cell fusion when expressed individually, suggesting that they were biologically fully active. This was indeed confirmed by incorporating one of the proteins into virions which thereby acquired a new host cell tropism, as will be reported elsewhere.


title: Occurrence of viruses in human stools in the Ahaggar (Alberia).

text: From October 1977 to May 1980, 243 stools collected in sedentary and semi-nomadic populations of the Ahaggar (Algerian Sahara) were examined using immunoelectronmicroscopy and tissue culture inoculation. Immunoelectronmicroscopy revealed the presence of rotaviruses in 8, coronaviruses in 26, adenoviruses in 5 and small round viruses in 4. Enteroviruses were isolated in tissue culture from 24 stools. Rotaviruses were present in the Ahaggar but were associated with little acute enteric disease. The high frequency of coronaviruses both in gastroenteritis patients and in patients without disease was surprising. The prevalence of enteroviruses in this hyperarid zone was similar to or higher than that found in noticeably more human countries. Further systematic bacterial, viral and parasitic examinations are required to clarify the role of the above viruses in the aetiology of gastroenteritis in this region.


title: Comparative trial of the canine parvovirus, canine distemper virus and canine adenovirus type 2 fractions of two commercially available modified live vaccines.

text: The results of vaccinating two groups of puppies with commercial vaccines, both of which claimed to provide adequate protection with a final vaccination at 10 weeks of age, were compared. Groups of 19 and 20 puppies with similar titres of maternally derived antibodies against canine parvovirus (cpv), canine distemper virus (cdv) and canine adenovirus type 2 (cav-2) at four weeks of age were vaccinated at six and 10 weeks of age and their responses to each vaccination were measured by comparing the titres against cpv, cdv and cav-2 in the serum samples taken immediately before the vaccination and four weeks later. After the vaccination at six weeks of age, all 19 of the puppies in group 1 had responded to cpv and cdv, and 14 had responded to cav-2; in group 2, 17 of the 20 had responded to cpv, 19 to cdv and 15 to cav-2. In both groups the puppies that did not respond to the first vaccination had responded serologically to cpv, cdv and cav-2 at 10 weeks of age.


title: Omental torsion in a captive polar bear (Ursus maritimus).

text: This is the first case report of an omental torsion in a polar bear (Ursus maritimus). A captive, 23-yr-old, 250-kg, intact female polar bear presented to the University of Minnesota Veterinary Medical Center with a 2-day history of lethargy, depression, and vomiting. Abdominal ultrasound identified large amounts of hyperechoic free peritoneal fluid. Ultrasound-guided abdominocentesis was performed and yielded thick serosanguinous fluid compatible with a hemoabdomen. An exploratory laparotomy revealed a large amount of malodorous, serosanguineous fluid and multiple necrotic blood clots associated with a torsion of the greater omentum and rupture of a branch of the omental artery. A partial omentectomy was performed to remove the necrotic tissue and the abdomen was copiously lavaged. The polar bear recovered successfully and is reported to be clinically well 6 mo later. This condition should be considered as a differential in bears with clinical signs of intestinal obstruction and hemoabdomen.


docid: kuginleu

title: Intracellular localization and protein interactions of the gene 1 protein p28 during mouse hepatitis virus replication.

text: Coronaviruses encode the largest replicase polyprotein of any known positive-strand RNA virus. Replicase protein precursors and mature products are thought to mediate the formation and function of viral replication complexes on the surfaces of intracellular double-membrane vesicles. However, the functions of only a few of these proteins are known. For the coronavirus mouse hepatitis virus (MHV), the first proteolytic processing event of the replicase polyprotein liberates an amino-terminal 28-kDa product (p28). While previous biochemical studies have suggested that p28 is associated with viral replication complexes, the intracellular localization and interactions of p28 with other proteins during the course of MHV replication have not been defined. We used immunofluorescence confocal microscopy to show that p28 localizes to viral replication complexes in the cytoplasm during early times postinfection. However, at late times postinfection, p28 localizes to sites of M accumulation distinct from the replication complex. Furthermore, by yeast two-hybrid and coimmunoprecipitation analyses, we demonstrate that p28 specifically binds to p10 and p15, two coronavirus replicase proteins of unknown function. Deletion mutagenesis experiments determined that the carboxy terminus of p28 is not required for its interactions with p10 and p15. These results suggest that p28 may play a part at the replication complex by interacting with p10 and p15. Moreover, our findings highlight a potential role for p28 at virion assembly sites.


title: Colds and influenza: a review of diagnosis and conventional, botanical, and nutritional considerations.

text: The common cold is the leading cause of doctor visits in the United States and annually results in 189 million lost school days. In the course of one year the U.S. population contracts approximately 1 billion colds. Influenza infection is still a leading cause of morbidity and mortality, accounting for 20-25 million doctor visits and 36,000 deaths per year in the United States. Conventional therapies for colds and flu focus primarily on temporary symptom relief and include over-the-counter antipyretics, anti-inflammatories, and decongestants. Treatment for influenza also includes prescription antiviral agents and vaccines for prevention. This article reviews the common cold and influenza viruses, presents the conventional treatment options, and highlights select botanicals (Echinacea spp., Sambucus nigra, larch arabinogalactan, Astragalus membranaceous, Baptisia tinctoria, Allium sativa, Panax quinquefolium, Eleutherococcus senticosus, Andrographis paniculata, olive leaf extract, and Isatis tinctoria) and nutritional considerations (vitamins A and C, zinc, high lactoferrin whey protein, N-acetylcysteine, and DHEA) that may help in the prevention and treatment of these conditions.


Identification of specific variations within the HE, S1, and ORF4 genes of bovine coronaviruses associated with enteric and respiratory diseases in dairy cattle.


docid: swoiypvt

title: Severe acute respiratory syndrome-global alert.


title: Paying the Price for PPE.

text: The early days of the COVID-19 pandemic threw the market for personal protective equipment (PPE) into chaos. So physicians and county medical societies across Texas found they had to go big or go home when it came to obtaining those critical supplies.


Bioethics, health, and inequality.


title: Daily Situation Report on Coronavirus disease (COVID-19) in Iran; March 23, 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 23, 2020.


title: Inpatient disposition classification for the creation of hospital surge capacity: a multiphase study.

text: BACKGROUND The ability to provide medical care during sudden increases in patient volume during a disaster or other high-consequence event is a serious concern for health-care systems. Identification of inpatients for safe early discharge (ie, reverse triage) could create additional hospital surge capacity. We sought to develop a disposition classification system that categorises inpatients according to suitability for immediate discharge on the basis of risk tolerance for a subsequent consequential medical event. METHODS We did a warfare analysis laboratory exercise using evidence-based techniques, combined with a consensus process of 39 expert panellists. These panellists were asked to define the categories of a disposition classification system, assign risk tolerance of a consequential medical event to each category, identify critical interventions, and rank each (using a scale of 1-10) according to the likelihood of a resultant consequential medical event if a critical intervention is withdrawn or withheld because of discharge. FINDINGS The panellists unanimously agreed on a five-category disposition classification system. The upper limit of risk tolerance for a consequential medical event in the lowest risk group if discharged early was less than 4%. The next categories had upper limits of risk tolerance of about 12% (IQR 8-15%), 33% (25-50%), 60% (45-80%) and 100% (95-100%), respectively. The expert panellists identified 28 critical interventions with a likelihood of association with a consequential medical event if withdrawn, ranging from 3 to 10 on the 10-point scale. INTERPRETATION The disposition classification system allows conceptual classification of patients for suitable disposition, including those deemed safe for early discharge home during surges in demand. Clinical criteria allowing real-time categorisation of patients are awaited.


title: [Genetic variation analysis of SARS coronavirus].

text: Severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) has been verified as the causative agent of a worldwide outbreak of SARS. Since the observed per-base replication error-rate of RNA polymerase is about 3 x 10(-5), RNA virus populations typically contain genetic variants that form a heterogeneous virus pool. This feature confers great adaptability on viruses and is partly responsible for current difficulties of viral disease prevention and control, such as of HIV and HCV infections. SARS-CoV, a newly-identified single-strand RNA virus, has been identified with common variable property of RNA viruses. In this study, genetic variation of SARS-CoV is analyzed based on available documented data and our study. It suggests that SARS-CoV is of similar mutation frequency and dynamic mutant distributions like other RNA viruses, though relatively less identified variation number than those causing chronic infections. In contrast, the viral spike (S) gene exhibits high variation, while nucleocapsid (N) gene shows relative sequence conservation. No obvious correlation is observed between viral variation and clinical disease manifestation from present analyzed data.


Aspirin and cognitive function.


title: Transcriptional profiling of acute cytopathic murine hepatitis virus infection in fibroblast-like cells.

text: Understanding the orchestrated genome-wide cellular responses is critical for comprehending the early events of coronavirus infection. Microarray analysis was applied to assess changes in cellular expression profiles during different stages of two independent, highly controlled murine hepatitis virus (MHV) infections in vitro. Fibroblast-like L cells were infected at high multiplicity in order to study the direct effects of a synchronized lytic coronavirus infection. Total RNA was harvested from MHV- or mock-infected L cells at 3, 5 and 6 h post-infection and hybridized to Affymetrix microarrays representing approximately 12,500 murine genes and expressed sequences. The expression data were compared to their respective mock-infected controls. Quantitative RT-PCR of selected transcripts was used to validate the differential expression of transcripts and inter-experiment reproducibility of microarray analysis. It was concluded that MHV-A59 infection in fibroblast-like cells triggers very few transcriptional cellular responses in the first 3 h of infection. Later, after having established a productive infection, a chemokine response is induced together with other cellular changes associated with RNA and protein metabolism, cell cycle and apoptosis. Interferon responses are not triggered during infection, although the L cells can be readily stimulated to produce interferon by dsRNA, a known potent inducer of interferon. Possibly, the interferon response is actively counteracted by a virus-encoded antagonist as has been described previously for other RNA viruses.


title: VenaTrac™ device for over-the-wire placement of chronic dialysis catheters.

text: PURPOSE The VenaTrac™ is designed for exchange and primary placement of chronic tunneled hemodialysis catheters over a wire. It occludes both lumens of the catheter for the purpose of reducing the risk of air embolism and blood loss. The purpose of this paper is to report our experience with the use of this device. METHODS A retrospective review of chronic hemodialysis dialysis catheters exchanges over a 30-month period was performed. Two hundred and ninety-seven over-the-wire catheter exchanges and 47 primary placements using the VenaTrac™ device were reviewed. In addition, 430 over-the-wire catheter exchanges without the use of the VenaTrac™ were reviewed. RESULTS No insertional complications or air embolism occurred when using the VenaTrac™. Symptomatic air embolism was documented in 5 out of 430 catheter exchanges performed without the use of VenaTrac™ (1.2%). CONCLUSIONS VenaTrac™ over-the-wire insertion device demonstrated safe and reliable use with no incidence of air embolism.


title: Antiherpesvirus activity in human sera and urines after administration of adenine arabinoside: in vitro and in vivo synergy of adenine arabinoside and arabinosylhypoxanthine in combination.

text: The minimum inhibitory concentration (MIC) of adenine arabinoside (ara-A) in rabbit kidney microtiter tissue cultures (RK-13) to a prototype strain of herpes simplex virus, type 1 (E115) based upon inhibition of cytopathic effects is 1.5 mug/ml. In this system, the MIC of arabinosylhypoxanthine (ara-Hx), the major in vivo metabolic derivative of ara-A, is 75 mug/ml. Inhibition of cytopathic effects of herpes simplex virus, type 1 (HSV-1) in microtiter wells of RK-13 cells varies directly with the concentrations of ara-A or ara-Hx, and inversely with residual HSV-1. The MIC of ara-A for HSV-1 in RK-13 cells is 5-20 times lower than similar measures with vero renal, mouse embryo, or human foreskin cultures. With RK-13 tissue cultures in microtiter plates, an assay for "ara-A equivalents" in human body fluids was developed which compares in sensitivity with high pressure liquid chromatography and has the advantage of simultaneously measuring combined antiherpesvirus effects of ara-A and its major metabolic derivative, ara-Hx. In vitro checkerboard studies in RK-13 cells confirmed that ara-A and ara-Hx in combination had antiviral effects which are synergistic. The total of the fractional MIC of ara-A plus ara-Hx in combination also varies inversely with residual HSV-1 in microtiter wells. Because virus adsorption is complete at 2 h before specimens to be tested are added in this assay, and because human interferon is not measured in rabbit cells, the antiviral assay is not affected by the presence of type-specific antiherpesvirus antibody or human interferon.Antiviral activity (AVA) was assayed as ara-A equivalents in sera and urines from 10 patients with serious herpesvirus infections who received 2.5-20 mg/kg daily of ara-A by intramuscular or intravenous routes. When a dosage schedule of 10 mg/kg per day or more was used, sustained concentrations of AVA that ranged from 0.8 to 14.4 mug/ml were found. When an inhibitor of adenosine deaminase (covidarabine) was not added to the specimens, mean serum concentrations were congruent with3.0 mug/ml (10 mg/kg per day, i.v.), and 4.1 mug/ml (20 mg/kg per day). However, in a single patient given 20 mg/kg of ara-A daily with covidarabine immediately added to the sera, the mean concentration of AVA was 12.9 mug/ml. Urines contained even higher AVA. Assays of 19 sera were performed both by microbiologic assay for AVA and by high pressure liquid chromatography for ara-A and ara-Hx. AVA was greater by microbiologic assay, and was greater than that which could be accounted for by stoichiometric chromatographic measures of ara-A and ara-Hx. These results with sera of treated patients are consistent both with the in vitro synergy of ara-A and ara-Hx found by checkerboard titrations, and with the beneficial responses to ara-A of patients with herpesvirus infections reported here and elsewhere.


title: Effect of time of exposure to rat coronavirus and Mycoplasma pulmonis on respiratory tract lesions in the Wistar rat.

text: The effects of time of exposure on the progression of pulmonary lesions in rats inoculated with Mycoplasma pulmonis and the rat coronavirus, sialodacryoadenitis virus (SDAV) were studied, using six groups of 18 SPF Wistar rats (n = 108). Rats were inoculated intranasally as follows: Group 1, sterile medium only; Group 2, sterile medium followed one week later by 150 TCID50 SDAV; Group 3, sterile medium followed by 10(5.7) colony forming units of M. pulmonis; Group 4, SDAV followed one week later by M. pulmonis; Group 5, M. pulmonis followed one week later by SDAV; Group 6, M. pulmonis followed two weeks later by SDAV. Six rats from each group were euthanized at one, two and three weeks after the final inoculation. In a separate experiment, six additional animals were inoculated in each of groups 3, 5 and 6 (n = 18) and were sampled at five weeks after they had received M. pulmonis. Bronchoalveolar lavage and quantitative lung mycoplasma cultures were conducted on two-thirds of the rats. Histopathological examination and scoring of lesion severity were performed on all animals. Based on the prevalence and extent of histopathological lesions, bronchoalveolar lavage cell numbers, neutrophil differential cell counts and the isolation of M. pulmonis, the most severe disease occurred in the groups that received both agents. There was no significant difference in lesion severity between the groups receiving both agents other than in those examined during the acute stages of SDAV infection. Based on these results, it is evident that SDAV enhances lower respiratory tract disease in Wistar rats whether exposure occurs at one week prior to or at various intervals following M. pulmonis infections.


Comparative aspects of togaviruses.


docid: txafcwfn

title: In vitro detection of apoptosis in monocytes/macrophages infected with human coronavirus.

text: Human coronavirus (HCoV) strain 229E infection, but not HCoV strain OC43 infection, of monocytes/macrophages from healthy donors and patients with multiple sclerosis in remission resulted in increased apoptosis, as measured by DNA changes and annexin V staining. Apoptosis correlated with the differential release of infectious virus. HCoV strain 229E titers were 10(3.5) to 10(6) 50% tissue culture-infective doses (TCID(50))/ml, and HCoV strain OC43 titers were only 10(1.2) to 10(2.7) TCID(50)/ml.


[Pandemic influenza].


title: Very late ischemic complications in flow-diverter stents: a retrospective analysis of a single-center series.

text: OBJECTIVE The authors evaluate the rate and discuss the pathomechanisms of very late (≥ 4-month) ischemic complications after flow-diverter stent (FDS) placement for intracranial aneurysms. METHODS The authors retrospectively reviewed the clinical data of the patients treated at Pitié-Salpêtrière Hospital between January 2010 and September 2014, who underwent FDS placement for intracranial aneurysm. The patients received dual-antiplatelet therapy (clopidogrel and aspirin) 5 days before and 3-6 months after the procedure and then aspirin alone for 6-9 months. An ischemic complication was defined as a sudden focal neurological deficit documented on diffusion-weighted images. RESULTS Eighty-six consecutive patients were included. Three (3.5%) patients treated with the Pipeline embolization device experienced a delayed acute ischemic stroke (2 cases of perforator/side-wall branch infarction and 1 case of thromboembolic stroke) with an average delay of 384 days (4 months, 20 months, and 13 months, respectively). The aneurysm locations were the left superior cerebellar artery, the right anterior choroid artery, and the left internal carotid artery (paraclinoid segment), respectively. The complications occurred after the patients had completed the antiaggregation protocol, except for Patient 1, who was receiving aspirin alone because of a spontaneous hematoma. At the acute phase, no in-stent thromboses were found on digital subtraction angiography. In Patient 2, the treated anterior choroid artery was occluded 20 months after the procedure. In Patient 3, a focal stenosis (approximately 40%) of the distal aspect of the FDS, probably caused by intimal hyperplasia, was seen. CONCLUSIONS Very late ischemic complications after FDS treatment were observed in 3.5% of the cases in the authors' series, some of which occurred as late as more than 1 year after placement.


title: Risk of feline infectious peritonitis in cats naturally infected with feline coronavirus.

text: A longitudinal survey of 820 cats in 73 households was conducted over a period of 6 years to establish the fate of pet cats that were seropositive after natural exposure to feline coronavirus (FCoV). In particular, their risk of developing feline infectious peritonitis (FIP) was determined. The seropositive cats were assigned to 1 of 3 groups: cats from households in which FIP had recently been diagnosed; cats from households in which FIP had not been diagnosed, but from which kittens had been relocated and subsequently died of FIP; and cats from households in which FIP had not been diagnosed. Cats in the first group were not at greater risk of developing FIP than were cats in the other 2 groups. Consequently, any household in which seropositive cats live must be considered a potential source of FCoV that can cause FIP. There was no evidence that the enhanced disease, which has been described after experimentally induced infection of seropositive cats, exists in nature. Thus, analysis of the survival of the seropositive cats over periods of up to 36 months indicated that their risk of developing FIP decreased with time, suggesting the development of immunity rather than increased susceptibility to disease. In addition, of 56 cats deemed to have been naturally reinfected because their anti-FCoV antibody titers decreased and subsequently increased, only 3 developed FIP.


title: The effect of supplementation of docosahexaenoic acid and arachidonic acid on visual acuity and neurodevelopment in larger preterm infants.

text: BACKGROUND Preterm infants may be born with deficits of both docosahexaenoic acid (DHA) and arachidonic acid (AA), but studies on supplementation of DHA and AA for preterm infants are limited. METHODS Preterm infants with a gestational age between 30 and 37 weeks who met all the inclusion criteria were enrolled in this double blind, randomized, comparative study. Infants over 2000 g body weight, over 32 weeks of gestation and in full feeding status would enter into the active intervention period of 6 months. Sixteen infants received Neoangelac Plus with AA and DHA supplementation. Eleven infants received Neoangelac without AA and DHA supplementation. The babies had scheduled physical examinations and their cognitive development, visual acuity, and vital signs to be checked. Adverse events were also recorded. RESULTS The mean Mental Development Index (MDI) scores for the supplementation and non-supplementation groups were 96.1 +/- 8.6 and 91.7 +/- 10.4 respectively at 6 months and 98.7 +/- 8.0 and 90.5 +/- 6.9 respectively at 1 year. The mean Physical Development Index (PDI) scores of these two groups were 102.2 +/- 10.5 and 95.4 +/- 13.2 respectively at 6 months and 98.0 +/- 5.8 and 86.7 +/- 11.1 respectively at 1 year. By repeated measures ANOVA, significant differences existed between groups for MDI and PDI (p = 0.020 and 0.008). However, there were no differences in visual acuity, physical examination variables or vital signs between these two groups. No obvious adverse effects were observed during the study period. CONCLUSION These results showed possible benefits in the neurodevelopment of larger preterm infants given formula supplemented with DHA and AA.


title: Association of mouse fibrinogen-like protein with murine hepatitis virus-induced prothrombinase activity.

text: Previously, we demonstrated induction of a unique macrophage prothrombinase during infection of BALB/cJ mice by mouse hepatitis virus strain 3 (MHV-3). By immunologic screening, a clone representing this prothrombinase was isolated from a cDNA library and sequenced. The sequence identified this clone as representing part of a gene, musfiblp, that encodes a fibrinogen-like protein. Six additional clones were isolated, and one clone, p11-3-1, encompassed the entire coding region of musfiblp. Murine macrophages did not constitutively express musfiblp but, when infected with MHV-3, synthesized musfiblp-specific mRNA. musfiblp mRNA induction was earlier and significantly greater in BALB/cJ than A/J macrophages. Prothrombinase activity was demonstrated when musfiblp was expressed from p11-3-1 in RAW 264.7 cells. These data suggest that musfiblp encodes the MHV-induced prothrombinase.


title: Prevalence of naturally occurring viral infections, Mycoplasma pulmonis and Clostridium piliforme in laboratory rodents in Western Europe screened from 2000 to 2003.

text: In this report prevalence rates of rodent viruses in laboratory animals are presented based on routine serological screening of mouse and rat colonies from European institutes. The prevalences found during the period 2000-2003 are compared with those reported for 1981-1984 and 1990-1993. It is shown that some infections were eliminated from laboratory animal colonies (e.g. K-virus and polyomavirus) by taking preventative measures whereas other infections such as mouse hepatitis virus and parvoviruses remained at a high rate. Further decreases in prevalence rates in the last 10 years were found for infections such as pneumonia virus of mice, reovirus type 3, Sendai virus, sialodacryoadenitis/rat coronavirus and Mycoplasma pulmonis. The introduction of new detection methods showed that mouse parvovirus and rat parvovirus, both members of the Parvoviridae family, remain a major threat to laboratory mice and rats. Guinea pig cytomegalovirus and para-influenza virus appeared to be the most prevalent agents among laboratory guinea pigs. The importance of a standardized, up-to-date screening programme is discussed.


title: Dihydrophenanthrenes from Bletilla formosana.

text: Three new dihydrophenanthrenes, 4-methoxy-9,10-dihydrophenanthrene-1,2,7-triol (1), 1-(4-hydroxybenzyl)-4,7-dimethoxy-9,10-dihydrophenanthrene-2-ol (2), and 1,3,6-tri(4-hydroxybenzyl)-4-methoxydihydrophenanthrene-2,7-diol (3) together with seven known phenanthrene derivatives, six known flavonoids, a bibenzyl and three phenolic compounds were isolated from the whole plant of Bletilla formosana. Their structures were elucidated by spectroscopic, mainly 2D NMR spectrometry and chemical methods.


title: The utility of preemptive mass influenza vaccination in controlling a sars outbreak during flu season.

text: During flu season, respiratory infections can cause non-specific influenza-like-illnesses (ILIs) in up to one-half of the general population. If a future SARS outbreak were to coincide with flu season, it would become exceptionally difficult to distinguish SARS rapidly and accurately from other ILIs, given the non-specific clinical presentation of SARS and the current lack of a widely available, rapid, diagnostic test. We construct a deterministic compartmental model to examine the potential impact of preemptive mass influenza vaccination on SARS containment during a hypothetical SARS outbreak coinciding with a peak flu season. Our model was developed based upon the events of the 2003 SARS outbreak in Toronto, Canada. The relationship of different vaccination rates for influenza and the corresponding required quarantine rates for individuals who are exposed to SARS was analyzed and simulated under different assumptions. The study revealed that a campaign of mass influenza vaccination prior to the onset of flu season could aid the containment of a future SARS outbreak by decreasing the total number of persons with ILIs presenting to the health-care system, and consequently decreasing nosocomial transmission of SARS in persons under investigation for the disease.


title: Inhibition of replication and infection of severe acute respiratory syndrome-associated coronavirus with plasmid-mediated interference RNA.

text: Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease caused by a novel coronavirus (SARS-CoV), which spread to over 30 countries in early 2003. Until recently, no specific vaccines and effective drugs have been available to protect patients from infection by this virus. To exploit a new strategy to fight this disease, we investigated the effect of interference RNA (RNAi) on the virus infection and replication with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT), plaque-forming, Western-blot and real-time PCR assays. Results showed that the plasmid-derived siRNAs targeting the non-structural protein 1 (NSP1) sequence of the SARS-CoV genome could specifically inhibit the expression of the NSP1 sequence and effectively suppress the replication and propagation of SARS-CoV in cultured Vero E6 cell lines. The expression of the Spike and Nucleoprotein genes of SARS-CoV at mRNA and protein levels in small interfering (si)RNA-expressing cells was significantly less than that in controls when analysed with PCR and Western-blot assays, 3 days post SARS-CoV infection. Our study provides strong evidence that the NSP1 sequence in the SARS-CoV genome is a valid target for RNAi and the effect of the siRNAs probably mainly resulted from effects on global reduction of subgenome synthesis and subsequent protein expression of SARS-CoV.


title: The minimal leak test technique for endotracheal cuff maintenance.

text: Endotracheal tube (ETT) cuff pressure management is an essential part of airway management in intubated and mechanically ventilated patients. Both under- and over-inflation of the ETT cuff can lead to patient complications, with an ideal pressure range of 20-30 cmH2O defined. A range of techniques are employed to ensure adequate ETT cuff inflation, with little comparative data. We performed an observational cross-sectional study in a tertiary metropolitan ICU, assessing the relationship between the minimal leak test and cuff manometry. Forty-five mechanically ventilated patients, over a three-month period, had ETT cuff manometry performed at the same time as their routine cuff maintenance (minimal leak test). Bedside nurse measurements were compared with investigator measurements. At the endpoint of cuff inflation, 20 of 45 patients (44%) had cuff pressures between 20 and 30 cmH2O; 11 of 45 patients (24%) had cuff pressures <20 cmH2O; 14 of 45 patients (31%) had cuff pressures ≥30 cmH2O. Univariate analysis demonstrated an association between both patient obesity and female gender requiring less ETT cuff volume (P=0.008 and P <0.001 respectively), though this association was lost on multivariate analysis. No association was demonstrated between any measured variables and cuff pressures. Inter-operator reliability in performing the minimal leak test showed no evidence of bias between nurse and investigators (Pearson coefficient = 0.897). We conclude the minimal leak test for maintenance of ETT cuffs leads to both over- and under-inflation, and alternative techniques, such as cuff manometry, should be employed.


Chinese QX strain of infectious bronchitis virus isolated in the UK.


title: Respiratory viruses and exacerbations of asthma in adults.

text: OBJECTIVE To study the role of respiratory viruses in exacerbations of asthma in adults. DESIGN Longitudinal study of 138 adults with asthma. SETTING Leicestershire Health Authority. SUBJECTS 48 men and 90 women 19-46 years of age with a mean duration of wheeze of 19.6 years. 75% received regular treatment with bronchodilators; 89% gave a history of eczema, hay fever, allergic rhinitis, nasal polyps, or allergies; 38% had been admitted to hospital with asthma. MAIN OUTCOME MEASURES Symptomatic colds and asthma exacerbations; objective exacerbations of asthma with > or = 50 l/min reduction in mean peak expiratory flow rate when morning and night time readings on days 1-7 after onset of symptoms were compared with rates during an asymptomatic control period; laboratory confirmed respiratory tract infections. RESULTS Colds were reported in 80% (223/280) of episodes with symptoms of wheeze, chest tightness, or breathlessness, and 89% (223/250) of colds were associated with asthma symptoms. 24% of 115 laboratory confirmed non-bacterial infections were associated with reductions in mean peak expiratory flow rate > or = 50 l/min through days 1-7 and 48% had mean decreases > or = 25 l/min. 44% of episodes with mean decreases in flow rate > or = 50 l/min were associated with laboratory confirmed infections. Infections with rhinoviruses, coronaviruses OC43 and 229E, influenza B, respiratory syncytial virus, parainfluenza virus, and chlamydia were all associated with objective evidence of an exacerbation of asthma. CONCLUSIONS These findings show that asthma symptoms and reductions in peak flow are often associated with colds and respiratory viruses; respiratory virus infections commonly cause or are associated with exacerbations of asthma in adults.


title: [Prognostic analysis of lung function and chest X-ray changes of 258 patients with severe acute respiratory syndrome in rehabilitation after discharge].

text: OBJECTIVE To analyze the clinical characteristics and prognostic changes of rehabilitating severe acute respiratory syndrome (SARS) patients through regular lung function tests and lung imaging studies after discharge and to retrospectively analyze the treatment data of these patients. METHODS 258 discharged SARS patients received regular SARS-Co virus IgG test, lung function test and chest X-ray and/or high resolution computerized tomography (HRCT) examination at General Hospital of PLA two months after discharge, and the treatment data of these patients were retrospectively analyzed. RESULTS 80.6% patients (208 of 258 patients) were positive for SARS-Co virus IgG. 21.3% patients (55 of 258 patients) showed lung diffusion abnormity (D(LCO) < 80%pred). Compared to 155 SARS-Co virus IgG positive patients without lung diffusion abnormity and 50 SARS-Co virus IgG negative patients, the 53 SARS-Co virus IgG positive patients with lung diffusion abnormity had longer fever course, higher dosages of glucocorticoid therapy, higher percentage of oxygen therapy and non-invasive ventilation. 51 of the 53 patients with lung diffusion abnormity received lung function test after one month, and the results of D(LCO) improved in 80.4% patients (41 of 51 patients). 40 of 51 patients with lung diffusion abnormity showed lung fibrosis, and the fibrosis decreased in 55% patients (22 of 40 patients) after one month. CONCLUSIONS This finding suggests that lung fibrosis caused by SARS mostly occurs in severe patients, and it can resolve spontaneously. D(LCO) may be more sensitive than HRCT in evaluating the fibrotic changes.