title: Molecular epidemiology of bovine coronavirus on the basis of comparative analyses of the S gene.

text: Bovine coronavirus (BCoV), a group 2 member of the genus Coronavirus in the family Coronaviridae, is an important pathogen in cattle worldwide. It causes diarrhea in adult animals (winter dysentery), as well as enteric and respiratory diseases in calves. The annual occurrence of BCoV epidemics in Sweden and Denmark led to this investigation, with the aim to deepen the knowledge of BCoV epidemiology at the molecular level. A total of 43 samples from outbreaks in both countries were used for PCR amplification and direct sequencing of a 624-nucleotide fragment of the BCoV S gene. Sequence comparison and phylogenetic studies were performed. The results showed (i) identical sequences from different animals in the same herds and from paired nasal and fecal samples, suggesting a dominant virus circulating in each herd at a given time; (ii) sequence differences among four outbreaks in different years in the same herd, indicating new introduction of virus; (iii) identical sequences in four different Danish herds in samples obtained within 2 months, implying virus transmission between herds; and (iv) that at least two different virus strains were involved in the outbreaks of BCoV in Denmark during the spring of 2003. This study presents molecular data of BCoV infections that will contribute to an increased understanding of BCoV epidemiology in cattle populations.


Learn from influenza epidemiology.


title: Defensive applications of gene transfer technology in the face of bioterrorism: DNA-based vaccines and immune targeting.

text: Gene transfer involves the introduction of an engineered gene into a person's cells with the expectation that the protein expressed from the gene will produce a therapeutic benefit. Strategies based on this principle have led to the approval of > 600 clinical trials and enrollment of approximately 3500 subjects worldwide in attempts to treat diseases ranging from cancer to AIDS to cystic fibrosis. While gene therapy has met with limited success and still has many hurdles to overcome before it sees wide application, it may be useful as a defensive strategy against bioterrorism agents including infectious microbes and toxins. Although many defensive strategies are possible, immunological strategies are currently the most developed and are being actively applied to the development of strategies against several of the most virulent potential bio-weapons. While most of these strategies are not yet ready for human application, DNA-based vaccines appear to be among the most promising in the fight against bioterrorism.


title: Tagging Recombinant Proteins to Enhance Solubility and Aid Purification.

text: Protein fusion technology has had a major impact on the efficient production and purification of individual recombinant proteins. The use of genetically engineered affinity and solubility-enhancing polypeptide "tags" has increased greatly in recent years and there now exists a considerable repertoire of these that can be used to solve issues related to the expression, stability, solubility, folding, and purification of their fusion partner. In the case of large-scale proteomic studies, the development of purification procedures tailored to individual proteins is not practicable, and affinity tags have therefore become indispensable tools for structural and functional proteomic initiatives that involve the expression of many proteins in parallel. Here, the rationale and applications of a range of established and more recently developed solubility-enhancing and affinity tags is described.


docid: gqgtgezg

title: Protective effect of Houttuynia cordata extract on bleomycin-induced pulmonary fibrosis in rats.

text: The present study aimed to examine the antioxidant properties of Houttuynia cordata (HC) and its protective effect on bleomycin-induced pulmonary fibrosis in rats. Results showed that aqueous extract of HC exhibited a different magnitude of antioxidant activities in all model systems tested. Although HC showed weaker free radical scavenging and xanthine oxidase inhibitory activity than vitamin E, its anti-lipid peroxidation activity in rat liver homogenate was close to that of vitamin E. In animal studies, HC significantly decreased the levels of superoxide dismutase, malondialdehyde, hydroxyproline, interferon-gamma, and tumor necrosis factor-alpha. However, an increase in the concentration of catalase was noted in the bronchoalveolar lavage fluid. HC also remarkably improved the morphological appearance of the lung of bleomycin-treated rats. These results suggest that HC possesses a protective effect against bleomycin-induced pulmonary fibrosis. Interestingly, this protective effect was more pronounced than that of vitamin E. In conclusion, the protective effect of HC on pulmonary fibrosis could be partly associated with the reduction of oxidative damage caused by bleomycin.


title: Identification of immunodominant epitopes on the membrane protein of the severe acute respiratory syndrome-associated coronavirus.

text: Similar to other coronaviruses, the membrane (M) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is a major transmembrane glycoprotein with multiple biological functions. To date, limited information is available about its antigenic properties. In this study, we identified two major immunodominant epitopes on the M protein located in the extreme N-terminal region (residues 1 to 31) and the interior C-terminal region (residues 132 to 161), respectively, by Pepscan analyses against convalescent-phase sera from SARS patients and antisera from virus-immunized mice and rabbits. Synthetic peptides M1-31 derived from the N-terminal epitope and M132-161 derived from the C-terminal epitope were highly reactive with all of the convalescent-phase sera from 40 SARS patients but not with 30 control serum samples from healthy blood donors, suggesting their potential application for serologic diagnosis of SARS. We showed that both peptides (M1-31 and M132-161) were able to induce high titers of antibody responses in the immunized rabbits, highlighting their antigenicity and immunogenicity. These findings provide important information for developing SARS diagnostics and vaccines.


title: Highlights and conclusions from the technical consultative meeting on novel coronavirus infection, Cairo, Egypt, 14-16 January 2013.

text: The emergence of a novel strain of coronavirus in the Arabian Peninsula raised a global health concern in 2012, partly because the majority of human infections were fatal and partly due to its presumed animal origin. An urgent meeting of scientific and public health experts was convened by WHO in January 2013 in view of the limited knowledge available on the epidemiological and natural history of infection with this novel virus. The meeting reviewed current evidence and identified critical knowledge gaps to improve better understanding of the public health risk associated with the virus so as to improve preparedness and to safeguard and protect global health.


title: Interpretation of diagnostic laboratory tests for severe acute respiratory syndrome: the Toronto experience.

text: BACKGROUND An outbreak of severe acute respiratory syndrome (SARS) began in Canada in February 2003. The initial diagnosis of SARS was based on clinical and epidemiological criteria. During the outbreak, molecular and serologic tests for the SARS-associated coronavirus (SARS-CoV) became available. However, without a "gold standard," it was impossible to determine the usefulness of these tests. We describe how these tests were used during the first phase of the SARS outbreak in Toronto and offer some recommendations that may be useful if SARS returns. METHODS We examined the results of all diagnostic laboratory tests used in 117 patients admitted to hospitals in Toronto who met the Health Canada criteria for suspect or probable SARS. Focusing on tests for SARS-CoV, we attempted to determine the optimal specimen types and timing of specimen collection. RESULTS Diagnostic test results for SARS-CoV were available for 110 of the 117 patients. SARS-CoV was detected by means of reverse-transcriptase polymerase chain reaction (RT-PCR) in at least one specimen in 59 (54.1%) of 109 patients. Serologic test results of convalescent samples were positive in 50 (96.2%) of 52 patients for whom paired serum samples were collected during the acute and convalescent phases of the illness. Of the 110 patients, 78 (70.9%) had specimens that tested positive by means of RT-PCR, serologic testing or both methods. The proportion of RT-PCR test results that were positive was similar between patients who met the criteria for suspect SARS (50.8%, 95% confidence interval [CI] 38.4%-63.2%) and those who met the criteria for probable SARS (58.0%, 95% CI 44.2%-70.7%). SARS-CoV was detected in nasopharyngeal swabs in 33 (32.4%) of 102 patients, in stool specimens in 19 (63.3%) of 30 patients, and in specimens from the lower respiratory tract in 10 (58.8%) of 17 patients. INTERPRETATION These findings suggest that the rapid diagnostic tests in use at the time of the initial outbreak lack sufficient sensitivity to be used clinically to rule out SARS. As tests for SARS-CoV continue to be optimized, evaluation of the clinical presentation and elucidation of a contact history must remain the cornerstone of SARS diagnosis. In patients with SARS, specimens taken from the lower respiratory tract and stool samples test positive by means of RT-PCR more often than do samples taken from other areas.


Applying rapid genome sequencing technologies to characterize pathogen genomes.


title: [Equipment for SARS control in patient care units and in units that process infected samples. 2003].

text: The information on severe acute respiratory syndrome (SARS) that has been gathered up to this point has made it possible to prepare recommendations concerning the equipment needed to protect health workers responsible for patient care and for processing potentially infected samples. Protecting such personnel is a key element in the strategy to control the spread of SARS. The needed equipment includes clothing; footwear; such protective devices as masks, safety glasses, and gloves; disinfectant solutions; laboratory equipment; and materials for obtaining and transporting samples. Prepared by the Western Pacific Regional Office of the World Health Organization, this list will help implement measures to contain the epidemic. The list gives the recommended quantities, specifications for the items, and possible alternatives for some items. This list of equipment is not exhaustive; it is intended to provide a small emergency supply for a period of 3 or 4 days for a single isolation unit with approximately 50 workers. To guarantee the availability of the equipment in the needed quantities, infection control authorities should take into account the number of isolation units needed and the length of time that containment measures will need to be in place. Adequate reserves should be available 24 hours a day, and plans should be made for rapid access to larger stocks in the event of a larger outbreak.


title: Outcomes and Complications of Simultaneous Laparoscopic Cystectomy and Laparoscopic Nephroureterectomy with Umbilical Reduced Port Surgery

text: Objective: In recent years, although reduced port surgeries (RPS) have been reported for many urological diseases, there have been no reports regarding simultaneous laparoscopic cystectomy and unilateral or bilateral nephroureterectomy with umbilical RPS. Therefore, the aim of this study was to evaluate outcomes and complications of simultaneous laparoscopic cystectomy and unilateral or bilateral nephroureterectomy with umbilical RPS. Methods: We performed a preliminary case series of 4 patients with synchronous upper urinary tract (UUT) tumor and invasive bladder cancer who underwent simultaneous laparoscopic cystectomy and unilateral or bilateral nephroureterectomy with umbilical RPS between 2014 and 2017 at our hospital. Demographic data, pathologic features, the surgical technique, and outcomes were retrospectively analyzed. Result: All 4 patients were men whose median age was 79 years (range 65-85 years) and median body mass index was 24.2 kg/m2 (range 21.5-27.3 kg/m2). The laparoscopic approach was technically successful in all 4 patients without the need for open conversion. The median total operative time was 434 minutes (range 372-481 minutes). The median estimated blood loss was 773 ml (range 153-923 ml), median interval to resuming oral intake was 2 days (range 1-7 days), and median hospital stay was 16 days (range 13-20 days). Conclusion: The reduced port approach is technically feasible in terms of many outcome measures, with significant cosmetic advantages. This method can be performed safely and recommended as a viable option for patients with concomitant UUT and bladder cancer.


SARS: prudence, not panic.


title: Isatin compounds as noncovalent SARS coronavirus 3C-like protease inhibitors.

text: A series of isatin derivatives were synthesized and tested against SARS CoV 3C-like protease. Substitutions at the N-1 and C-5 positions were examined to elucidate the differences in substrate binding sites of the rhinovirus 3C protease and SARS CoV 3C-like protease. Compound 5f shows significant inhibition with an IC(50) of 0.37 microM. Further study showed that, unlike the irreversible covalent binding of isatin derivatives to human rhinovirus 3C protease, the compounds tested in this study are all noncovalent reversible inhibitors.


title: Polypeptides and functions of antigens from human coronaviruses 229E and OC43.

text: Coronaviruses possess three major size classes of polypeptides as judged by molecular weight: approximately 180,000, approximately 50,000, and approximately 23,000. Human coronaviruses 229E and OC43 possess not only three similar size classes of polypeptides but also three distinct antigens, none of which cross-react with the heterologous strain. Polypeptides separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were reacted in rocket immunoelectrophoresis with antiserum monospecific to each of the three strain-specific antigens (excised precipitin lines from crossed immunoelectrophoresis profiles were used for immunogens). Monospecific antiserum with neutralizing ability reacted with a polypeptide of 186,000 daltons for 229E and a polypeptide of 190,000 daltons for OC43. The antigen which elicited neutralizing antibody response was located at the surface, associated with the corona of the virion, glycosylated, and bound by concanavalin A. Another less prominent surface antigen was represented by size classes of 23,000 daltons for 229E and 24,000 for OC43. The core antigens of the viruses had molecular weights of 49,000 and 229E and 52,000 and OC43 virus. Thus, the molecular weights and functions of the antigens of human coronaviruses are similar to those of animal coronaviruses. The polypeptides of coronaviruses 229E and OC43 are nearly identical as judged by molecular weight, but the similar polypeptides of the two viruses represent different immunological specificities.


title: Changes in locations of crossover sites over time in de novo generated RNA recombinants.

text: Recombinant RNAs generated in plants 3 weeks postinoculation with turnip crinkle virus (TCV) genomic RNA and an associated satellite RNA, sat-RNA D, have a majority of TCV crossover sites in a 24-nucleotide repeat (motif IIIA/IIIB) that forms part of a stable hairpin (Carpenter et al., 1995, J. Mol.Biol. 245, 608-622). To determine if parameters other than nucleotide sequence in the crossover region affect junction site selection, recombinants were assayed at various times postinoculation of plants and protoplasts. Populations of recombinants became progressively shorter in plants and larger in protoplasts. Levels of inoculated transcript and age of the plant were not substantial factors in the shifts in crossover site locations. The two most commonly cloned recombinant species were not amplified to detectable levels in protoplasts, suggesting that these molecules are not viable templates for replication. These results suggest that recombination between sat-RNA D and TCV is a very frequent event, and populations of recombinants are likely generated de novo in each infected cell and represent the original recombinant molecules rather than progeny of such molecules. Therefore, factors other than simple selection for recombinants that are more fit to replicate are probably responsible for the differences in junction sites in populations of sat-RNA D/TCV recombinants.


Occult respiratory viral infections in coronial autopsies: a pilot project.


title: Risk factors for SARS infection within hospitals in Hanoi, Vietnam.

text: We investigated a nosocomial infection of severe acute respiratory syndrome (SARS) in Vietnam in 2003 and attempted to identify risk factors for SARS infection. Of the 146 subjects who came into contact with SARS patients at Hospital A, 43 (29.5%) developed SARS, and an additional 16 (11%) were asymptomatic but SARS-coronavirus (CoV) seropositive. The asymptomatic infection rate accounted for 15.5% of the total number of infected patients at Hospital A, which was higher than that of 6.5% observed at Hospital B, to where all patients from Hospital A were eventually transported (P<0.05). At Hospital A, the risk for developing SARS was 12.6 times higher in individuals not using a mask than in those using a mask. The SARS epidemic in Vietnam resulted in numerous secondary infections due to its unknown etiology and delayed recognition at the beginning of the epidemic. The consistent and proper use of a mask was shown to be crucial for constant protection against infection with SARS.


Erratum To: Accounting for Behavioral Responses during a Flu Epidemic Using Home Television Viewing.


title: Bactericidal activity of a Ce-promoted Ag/AlPO4 catalyst using molecular oxygen in water.

text: The catalytic inactivation of Escherichia coli in water by a cerium (Ce)-promoted silver-loaded aluminum phosphate (Ag/ AlPO4) catalyst using molecular oxygen was investigated. With optimum Ce content, the Ag(Ce)/AlPO4 catalyst exhibited strong bactericidal activity. The process of decomposition of the cell wall and cell membrane was directly observed by TEM. The different morphological changes of E. coli cells treated with the Ag(Ce)/AlPO4 catalyst and those treated with Ag+ suggested that the Ag+ eluted from the catalyst surface did not play an important role during the bactericidal process. Results of DMPO spin-trapping measurements by electron spin resonance (ESR) indicated the formation of the reactive oxygen species (ROS) *OH and *O2-, which caused the considerable bactericidal activity. The formation of H2O2 acted as an important intermediate; this was confirmed by addition of catalase as the scavenger. A possible catalytic oxidation bactericidal mechanism using molecular oxygen was proposed for the Ag(Ce)/AlPO4 catalyst.


title: Infection prevention and control in blood purification centers during the COVID-19 epidemic: a single institution experience from Zhejiang, China.

text: not available.


Evidence of torovirus infection in diarrhoeic cattle.


title: Hybridoma antibodies to the murine coronavirus JHM: characterization of epitopes on the peplomer protein (E2).

text: A panel of hybridoma antibodies that react with the surface peplomer glycoprotein (E2) of the murine coronavirus JHM were produced to characterize major antigenic domains associated with functions related to virulence. Three groups of hybridoma antibodies were differentiated by immunoprecipitation of lysates from JHM-infected cells. One group precipitated the virion structural proteins gp170 and gp98 together with the intracellular form of E2, gp150. A second group reacted with gp98 and gp150, and a third group precipitated gp150 only. Competition assays with biotinylated hybridoma antibodies allowed the definition of at least six different epitope groups. Only those antibodies which immunoprecipitated both gp170 and gp98 neutralized infectivity, inhibited cell fusion and protected infected rats against acute disease. Another class of antibodies binding to gp170 and gp98 also neutralized JHM virus, but did not inhibit fusion and did not protect against disease. Antibodies that immunoprecipitated gp150 and gp98 revealed only weak neutralization and did not inhibit cell fusion or protect animals. Four epitopes were defined by antibodies that immunoprecipitated gp150, but revealed no biological activity. These data indicate that the site responsible for cell fusion is associated with an epitope group carried by gp170 and gp98. Neutralizing antibodies bind to this and another epitope. Furthermore, protection of JHM-infected rats against acute disease requires both inhibition of cell fusion and neutralization of virus.


title: Biphasic immune responses of cats under controlled infection with a feline enteric coronavirus-79-1683 strain.

text: Kittens inoculated orally with 10(2) PFU of feline enteric coronavirus developed no antibody to the virus despite the repeated challenges. However, they developed antibody for a long period with 5 x 10(3)-1.6 x 10(5) (mean 3 x 10(4)) and with 2.5 x 10(3)-2 x 10(4) (mean 6 x 10(3)) immunoperoxidase antibody titer when they were challenged with 10(5) and 10(3) PFU of virus following previous challenges, respectively. Viremia was found when kittens were inoculated with 10(5) PFU of virus, but not with 10(3) PFU of virus. The dose of 10(3) PFU of virus seemed to be a lower limit to establish infection. These results indicate that local infection induces a low antibody response and systemic infection induces a high antibody response.


title: Refining estimates of public health spending as measured in national health expenditure accounts: the Canadian experience.

text: The recent focus on public health stemming from, among other things, severe acute respiratory syndrome and avian flu has created an imperative to refine health-spending estimates in the Canadian Health Accounts. This article presents the Canadian experience in attempting to address the challenges associated with developing the needed taxonomies for systematically capturing, measuring, and analyzing the national investment in the Canadian public health system. The first phase of this process was completed in 2005, which was a 2-year project to estimate public health spending based on a more classic definition by removing the administration component of the previously combined public health and administration category. Comparing the refined public health estimate with recent data from the Organization for Economic Cooperation and Development still positions Canada with the highest share of total health expenditure devoted to public health than any other country reporting. The article also provides an analysis of the comparability of public health estimates across jurisdictions within Canada as well as a discussion of the recommendations for ongoing improvement of public health spending estimates. The Canadian Institute for Health Information is an independent, not-for-profit organization that provides Canadians with essential statistics and analysis on the performance of the Canadian health system, the delivery of healthcare, and the health status of Canadians. The Canadian Institute for Health Information administers more than 20 databases and registries, including Canada's Health Accounts, which tracks historically 40 categories of health spending by 5 sources of finance for 13 provincial and territorial jurisdictions. Until 2005, expenditure on public health services in the Canadian Health Accounts included measures to prevent the spread of communicable disease, food and drug safety, health inspections, health promotion, community mental health programs, public health nursing, as well as all the costs for the general administration of government health departments.


title: Polytope vaccines for the codelivery of multiple CD8 T-cell epitopes.

text: Vaccines against a number of diseases, including HIV, Epstein Barr virus, malaria and several cancers, are believed to require the coinduction of multiple alphabeta CD8+ cytotoxic T-lymphocyte responses that are directed towards a number of different target antigens. The difficulties associated with making large recombinant vaccines that contain numerous antigens has led to the development of alphabeta CD8+ cytotoxic T-lymphocyte polyepitope or polytope vaccine approach, where multiple (usually 8-10 amino acids long) alphabeta CD8+ cytotoxic T-lymphocyte epitopes, derived from several antigens are conjoined into single artificial constructs. Such polytope constructs can be delivered using a number of different vaccine vector modalities with each epitope in the construct emerging as individually immunogenic.


Rethinking global health training in North America.


title: Two cases of severe obstructive pneumonia associated with an HKU1-like coronavirus.

text: BACKGROUND During the last few years a number of previously undescribed viruses, including human metapneumovirus, coronaviruses SARS, NL63 and HKU1, and bocavirus, were identified in nasopharyngeal samples from patients with signs of respiratory infections. These viruses may cause mild to life-threatening infections. OBJECTIVES Nasopharyngeal samples from hospitalized pediatric patients with respiratory disease were analysed for the presence of coronaviruses and other well known and newly identified respiratory viruses. RESULTS Two clinical cases of a severe obstructive pneumonia, which were associated with the presence of RNA of a novel variant (subtype) of HKU1 coronavirus in the nasopharyngeal aspirates, were identified. DISCUSSION The detection of a HKU1-like coronavirus in pediatric patients in the current study complement the most recent independent finding of similar or closely related coronaviruses in patients with respiratory diseases in France (Vabret et al. 2006) and Norway (Jonassen et al., see accompanying manuscript). These observations indicate a wide dissemination of HKU1-like coronaviruses in Europe.


docid: qmjdyrdl

title: Do we need genomic research for the prevention of common diseases with environmental causes?

text: Concerns have been raised about the value of genomic research for prevention and public health, especially for complex diseases with risk factors that are amenable to environmental modification. Given that gene-environment interactions underlie almost all human diseases, the public health significance of genomic research on common diseases with modifiable environmental risks is based not necessarily on finding new genetic "causes" but on improving existing approaches to identifying and modifying environmental risk factors to better prevent and treat disease. Such applied genomic research for environmentally caused diseases is important, because 1) it could help stratify disease risks and differentiate interventions for achieving population health benefits; 2) it could help identify new environmental risk factors for disease or help confirm suspected environmental risk factors; and 3) it could aid our understanding of disease occurrence in terms of transmission, natural history, severity, etiologic heterogeneity, and targets for intervention at the population level. While genomics is still in its infancy, opportunities exist for developing, testing, and applying the tools of genomics to clinical and public health research, especially for conditions with known or suspected environmental causes. This research is likely to lead to population-wide health promotion and disease prevention efforts, not only to interventions targeted according to genetic susceptibility.


title: In vivo RNA-RNA recombination of coronavirus in mouse brain.

text: RNA-RNA recombination between different strains of the murine coronavirus mouse hepatitis virus (MHV) occurs at a very high frequency in tissue culture. To demonstrate that RNA recombination may play a role in the evolution and pathogenesis of coronaviruses, we sought to determine whether MHV recombination could occur during replication in the animal host of the virus. By using two selectable markers, i.e., temperature sensitivity and monoclonal antibody neutralization, we isolated several recombinant viruses from the brains of mice infected with two different strains of MHV. The recombination frequency was very high, and recombination occurred at multiple sites on the viral RNA genome. This finding suggests that RNA-RNA recombination may play a significant role in natural evolution and neuropathogenesis of coronaviruses.


Emerging and new infectious diseases.


title: Viral-induced rhinitis.

text: Upper respiratory viruses cause self-limited illness characterized by acute rhinitis. In rhinovirus colds the symptoms are thought to be caused by the host response rather than viral damage of the nasal epithelium. Rhinovirus triggers an inflammatory cascade, evidenced by the presence of inflammatory mediators (e.g., IL-8) and proinflammatory cytokines (e.g., kinins) in nasal secretions, which results in symptomatic illness. In contrast to rhinovirus and coronavirus, which do not cause discernible epithelial damage, influenza virus and adenovirus do damage the nasal epithelium. Appropriate antiviral therapy will depend on the causative virus. Treatment of rhinovirus colds may require an antiviral agent (e.g., interferon alpha) in combination with antiinflammatory medication.


title: Plasma vitamin C and tocotrienols changes in response to dietary supplementation among young male adults.

text: BACKGROUND AND AIMS Intake of the antioxidant vitamins C and E lowers the oxidative stress. The study aimed to determine plasma concentrations of vitamin C and tocotrienols after supplementation of both vitamins in young male adults. MATERIALS AND METHODS A total of 64 police recruits were randomly assigned to one of these groups: (a) 500 mg vitamin C (Vitamin C), (b) 200 mg Tocovid (Tocotrienol), (c) combination of 500 mg vitamin C and 200 mg Tocovid (Combination) or (d) placebo (Placebo) for eight-weeks of supplementation followed by six-week washout period. RESULTS In Combination group, mean plasma vitamin C concentration significantly increased from baseline 2.86 +/- 1.19 mg/L to 10.37 +/- 1.29 mg/L and 15.63 +/- 1.27 mg/L after four- and eight-week supplementation, respectively. The corresponding figures for alpha-, delta- and gamma-tocotrienols were 9.9 +/- 2.5 ng/ml to 104.1 +/- 19.8 ng/ml and 112.8 +/- 38.0 ng/ml; 2.5 +/- 0.9 ng/ml to 29.9 +/- 7.0 ng/ml and 17.9 +/- 4.7 ng/ml; 19.2 +/- 3.1 ng/ml to 75.2 +/- 24.1 ng/ml and 161.7 +/- 49.9 ng/ml, respectively. In Vitamin C group, plasma vitamin C concentrations were significantly increased. Conversely, concentration of plasma vitamin C in Tocotrienol group increased from baseline of 2.72 +/- 0.20 mg/L to 6.80 +/- 0.63 mg/L and 8.9 +/- 0.77 mg/L respectively. Plasma concentrations of alpha-, delta- and gamma-tocotrienols in this group were significantly elevated. After 6-week washout period, all the elevated concentrations returned to basal levels. CONCLUSION The study showed a good bioavailability of these vitamins and increment due to supplementation.


title: Utilising a defective IBV RNA for heterologous gene expression with potential prophylactic application.

text: Based on the natural ability of coronaviruses to undergo homologous RNA recombination, we are working to produce infectious bronchitis virus (IBV) recombinants using RNA generated from recombinant fowlpox viruses (FPV). The aim is to replace the spike (S) gene of an existing IBV vaccine strain with the S gene of a heterologous strain. CD-61 is an IBV defective RNA (D-RNA) derived from a naturally occurring IBV D-RNA (CD-91). CD-61 D-RNA is being investigated as an RNA vector for the expression of heterologous genes. T7-derived RNA transcripts of CD-61 can be replicated and passaged in the presence of helper virus, following electroporation into IBV-infected cells. CD-61 cDNA was modified by the addition of the hepatitis delta virus ribozyme plus T7 terminator downstream of the 3'UTR. This allowed the synthesis of discreet RNA transcripts. The complete cassette was cloned into an FPV transfer vector (pEFL10) for generating recombinant fowlpox viruses. FPV/CD-61 recombinants will be assessed for D-RNA production in IBV-infected cells. The luciferase reporter gene sequence has been inserted into the modified CD-61, under the control of the IBV transcription associated sequence (TAS) from gene 5. Luciferase has been successfully expressed from CD-61 in helper virus-infected cells.


title: Infection control, ethics and accountability.

text: Health care-associated infections (HAIs) are a major clinical and economic problem in Australian hospitals, and a significant proportion are preventable. HAIs are the result of complex environmental, microbiological, pathological, behavioural and organisational factors, and prevention requires a multifaceted ("bundled") approach, including appropriate policies, educational programs for health care workers, and adequate resources to implement them effectively. Failure to protect patients from avoidable harm, including HAIs, has significant ethical implications; it often reflects both organisational systems failure and non-compliance of health care workers with evidence-based policies, including hand hygiene. If implemented with appropriate safeguards, infection control "bundles" that include sanctions for poor compliance with hand hygiene and other infection control policies, will achieve sustained improvements where previous approaches have failed.


title: Analysis of endovascular treatment of ruptured aneurysms of the middle cerebral artery compared to other anatomical locations.

text: OBJECTIVE The objective of this article is to compare the results of endovascular treatment of ruptured middle cerebral artery (MCA) aneurysms with ruptured aneurysms of other anatomic locations. METHODS Fifty consecutive ruptured aneurysms of the MCA and 209 aneurysms at other anatomical locations were selected retrospectively. We compared epidemiological, clinical and radiological variables, prognosis and complications. RESULTS The MCA aneurysms had a greater size and a poor dome/neck ratio. There were no significant differences in endovascular technique complications, occlusion rate or rebleeding between the two groups (p > 0.1). There were no significant differences in the mortality and number of dependent patients after one month. CONCLUSION The endovascular treatment of ruptured MCA aneurysms without hematoma is as safe and effective as other aneurysm localizations. Complication rates, occlusion rates and rebleeding of ruptured MCA aneurysms are comparable to other locations.


title: Haemodynamic response associated with both ictal and interictal epileptiform activity using simultaneous video electroencephalography/near infrared spectroscopy in a within-subject study.

text: This paper reports the findings from a pilot study of four patients with medically refractory epilepsy undergoing pre-surgical evaluation with ages ranging from 5 to 17 years. Video electroencephalography recordings and data from a near infrared spectroscopy cerebral/somatic oximeter were gathered and related to electrographic seizure onset and offset as determined by a paediatric epileptologist. All four patients showed haemodynamic changes associated with epileptiform activities. The increased blood flow clearly coincided with epileptiform activity and continued to increase as the epileptiform activity built up. Regional cerebral oxygen saturation increased in the epileptogenic focus, perhaps due to loss of cerebrovascular autoregulation. These findings reinforce that near infrared spectroscopy can potentially be used in a wide spectrum of patients with epilepsy regardless of the underlying brain pathology.


title: Fundamental factors affecting upper-room ultraviolet germicidal irradiation - part II. Predicting effectiveness.

text: Compared with increasing outdoor air ventilation rate, upper-room ultraviolet germicidal irradiation (UVGI) is an attractive technology for lowering the indoor concentration of airborne microorganisms and thereby reducing the risk of airborne transmission of disease. With relatively modest vertical air circulation, most of the air in a room can be irradiated in a relatively brief time period without noise or significant power consumption. The hypothesis tested in this study is that the efficacy of upper-room UVGI to inactivate or kill airborne infectious microorganisms can be determined from an index of UVGI effectiveness, a dimensionless parameter designed to characterize adequacy of vertical air circulation, amount of UVGI provided, and their interaction. This index of effectiveness, which is determined independently of microbiological testing, was found to correlate well with experimental measurements made in a room-size chamber. A comparison of two other dimensionless parameters - the irradiation number and mixing number, from which effectiveness index is calculated - provides insight into whether the quantity of UV provided to the upper room or the intensity of the vertical air circulation is the controlling factor for effective application of upper-room UVGI. The irradiation number is calculated from the UV power output of the fixture(s), a parameter that is fixture specific and much easier to measure than mean fluence rate. An equation was also developed that relates UV fixture power output to mean fluence rate in either the irradiated zone or the entire room. In addition, reductions in viable microorganism concentration due to UVGI predicted from a two-box model are compared with experimental measurements.


title: [Rapid point-of-care serology testing for sars-cov-2].

text: Increasing evidence indicates immunity against severe acute respiratory syndrome coronavirus 2 (sars-cov-2) after covid-19, but it remains unclear for how long the protection remains. Serology testing seems to have a higher sensitivity than molecular diagnostics from 8 days after onset of symtoms, and should be part of risk assessment and epidemiological studies of COVID-19. The performance of commercial serological point-of-care (POC) lateral flow tests are highly manufacturer-dependant. Low sensitivity increases the risk of false negative results and could result in unnecessary quarantine of test persons with developed antibodies. Low specificity increases the risk of false positive results and could lead to false assumptions of immunity. Carefully selected serological POC tests for sars-cov-2 can be used in large scale testing but should only be used by licensed medical staff able to understand their limitations and interpret the results.


title: Structure of aminopeptidase N from Escherichia coli suggests a compartmentalized, gated active site.

text: Aminopeptidase N from Escherichia coli is a major metalloprotease that participates in the controlled hydrolysis of peptides in the proteolytic pathway. Determination of the 870-aa structure reveals that it has four domains similar to the tricorn-interacting factor F3. The thermolysin-like active site is enclosed within a large cavity with a volume of 2,200 A(3), which is inaccessible to substrates except for a small opening of approximately 8-10 A. The substrate-based inhibitor bestatin binds to the protein with minimal changes, suggesting that this is the active form of the enzyme. The previously described structure of F3 had three distinct conformations that were described as "closed," "intermediate," and "open." The structure of aminopeptidase N from E. coli, however, is substantially more closed than any of these. Taken together, the results suggest that these proteases, which are involved in intracellular peptide degradation, prevent inadvertent hydrolysis of inappropriate substrates by enclosing the active site within a large cavity. There is also some evidence that the open form of the enzyme, which admits substrates, remains inactive until it adopts the closed form.


title: Stability of Middle East respiratory syndrome coronavirus (MERS-CoV) under different environmental conditions.

text: The stability of Middle East respiratory syndrome coronavirus (MERS-CoV) was determined at 20°C--40% relative humidity (RH); 30°C--30% RH and 30°C--80% RH. MERS-CoV was more stable at low temperature/low humidity conditions and could still be recovered after 48 hours. During aerosolisation of MERS-CoV, no decrease in stability was observed at 20°C--40% RH. These data suggest the potential of MERS-CoV to be transmitted via contact or fomite transmission due to prolonged environmental presence.


title: [An investigation on nosocomial infection with severe acute respiratory syndrome in health-care workers at 13 key hospitals in Guangdong Province].

text: OBJECTIVE To analysis the risk factors influencing nosocomial infection of severe acute respiratory syndrome (SARS) in health-care workers and to evaluate effectiveness of its control and preventive measures in 13 key hospitals caring for SARS patients. METHODS Number of SARS patients, clinical conditions of them, its attack rate in health-care workers, and characteristics of hospitals, including their environment, isolating measures, etc. were investigated at the 13 hospital in Guangzhou to analyze the risk factors influencing nosocomial infection of SARS and its attack rates in health-care workers before and after implementation of preventive measures and to evaluate their effectiveness. RESULTS Totally, 841 patients with SARS were treated at the 13 hospitals in Guangzhou and 285 health-care workers caring for them infected nosocomially. Attack rate in health-care workers was higher at general hospitals, hospital accepting more cases in critical conditions and hospitals with poor precautious measures, and lower in hospitals with isolated wards or areas, or department of infection, specially caring for SARS patients, and those with effective intervention measures to prevent secondary infection. CONCLUSION Nosocomial infection of SARS in health-care workers was affected by clinical condition of SARS patients, characteristics and environment of hospitals and their personal protective measures adopted.


title: Isolation and identification of infectious bronchitis virus from chickens in Sichuan, China.

text: A nonhemagglutinating virus was isolated from kidneys and lungs of chickens suspected of having infectious bronchitis infection. Specific-pathogen-free embryonated chicken eggs were used as the cultural system. With the use of the ciliary activity of chicken embryo tracheal organ cultures as indicator system, the physicochemical properties of one of the isolated strains (SAIB3) were shown to be similar to infectious bronchitis virus (IBV) strain M41 (standard strain); whereas electron microscopy of the isolate showed coronavirus particles. Virus-neutralization tests were performed in tracheal organ cultures to compare the serotypes of five IBV isolates and six known IBV strains on the basis of reciprocal neutralization titers and euclidean distance. The cross-neutralization pattern indicated that one isolate was of the T-strain IBV serotype, another of the M41 IBV serotype, while others had partial serotype relationship to M41 and T-strains of IBV.


title: A novel transglutaminase substrate from Streptomyces mobaraensis inhibiting papain-like cysteine proteases.

text: Transglutaminase from Streptomyces mobaraensis is an enzyme of unknown function that cross-links proteins to high molecular weight aggregates. Previously, we characterized two intrinsic transglutaminase substrates with inactivating activities against subtilisin and dispase. This report now describes a novel substrate that inhibits papain, bromelain, and trypsin. Papain was the most sensitive protease; thus, the protein was designated Streptomyces papain inhibitor (SPI). To avoid transglutaminase-mediated glutamine deamidation during culture, SPI was produced by Streptomyces mobaraensis at various growth temperatures. The best results were achieved by culturing for 30-50 h at 42 degrees C, which yielded high SPI concentrations and negligibly small amounts of mature transglutaminase. Transglutaminasespecific biotinylation displayed largely unmodified glutamine and lysine residues. In contrast, purified SPI from the 28 degrees C culture lost the potential to be cross-linked, but exhibited higher inhibitory activity as indicated by a significantly lower Ki (60 nM vs. 140 nM). Despite similarities in molecular mass (12 kDa) and high thermostability, SPI exhibits clear differences in comparison with all members of the wellknown family of Streptomyces subtilisin inhibitors. The neutral protein (pI of 7.3) shares sequence homology with a putative protein from Streptomyces lavendulae, whose conformation is most likely stabilized by two disulfide bridges. However, cysteine residues are not localized in the typical regions of subtilisin inhibitors. SPI and the formerly characterized dispase-inactivating substrate are unique proteins of distinct Streptomycetes such as Streptomyces mobaraensis. Along with the subtilisin inhibitory protein, they could play a crucial role in the defense of vulnerable protein layers that are solidified by transglutaminase.


title: Adverse childhood experiences reported by adults --- five states, 2009.

text: Adverse childhood experiences (ACEs) include verbal, physical, or sexual abuse, as well as family dysfunction (e.g., an incarcerated, mentally ill, or substance-abusing family member; domestic violence; or absence of a parent because of divorce or separation). ACEs have been linked to a range of adverse health outcomes in adulthood, including substance abuse, depression, cardiovascular disease, diabetes, cancer, and premature mortality. Furthermore, data collected from a large sample of health maintenance organization members indicated that a history of ACEs is common among adults and ACEs are themselves interrelated. To examine whether a history of ACEs was common in a randomly selected population, CDC analyzed information from 26,229 adults in five states using the 2009 ACE module of the Behavioral Risk Factor Surveillance System (BRFSS). This report describes the results of that analysis, which indicated that, overall, 59.4% of respondents reported having at least one ACE, and 8.7% reported five or more ACEs. The high prevalence of ACEs underscores the need for 1) additional efforts at the state and local level to reduce and prevent child maltreatment and associated family dysfunction and 2) further development and dissemination of trauma-focused services to treat stress-related health outcomes associated with ACEs.


title: The history of ultraviolet germicidal irradiation for air disinfection.

text: Public health concerns such as multi- and extensive drug-resistant tuberculosis, bioterrorism, pandemic influenza, and severe acute respiratory syndrome have intensified efforts to prevent transmission of infections that are completely or partially airborne using environmental controls. One such control, ultraviolet germicidal irradiation (UVGI), has received renewed interest after decades of underutilization and neglect. With renewed interest, however, come renewed questions, especially regarding efficacy and safety. There is a long history of investigations concluding that, if used properly, UVGI can be safe and highly effective in disinfecting the air, thereby preventing transmission of a variety of airborne infections. Despite this long history, many infection control professionals are not familiar with the history of UVGI and how it has, and has not, been used safely and effectively. This article reviews that history of UVGI for air disinfection, starting with its biological basis, moving to its application in the real world, and ending with its current status.


title: Abdominal Wall Endometriosis on the Right Port Site After Laparoscopy: Case Report and Literature Review.

text: BACKGROUND Endometriosis can be intrapelvic or, rarely, extrapelvic. Endometriosis involving the rectus abdominis muscle on the trocar port site is a rare event; until now, only 16 cases have been reported in the literature. The majority of cases were associated with previous abdominal surgery such as diagnostic laparoscopy, cyst excision, appendectomy, myomectomy, or cholecystectomy. We review all the reported cases of this unusual form of extrapelvic endometriosis. CASE REPORT We report a new case of abdominal wall endometriosis at the trocar port site in the rectus abdominis muscle in a woman who had undergone 2 laparoscopies for endometriosis in the 3 years before coming to our attention. The diagnosis was made by sonography. We performed a surgical resection of the lesion with a free macroscopic margin of 5-10 mm. CONCLUSION Endometriosis should be considered in the differential diagnosis of any abdominal swelling. In our experience, surgery is the treatment of choice.


title: Transgenic plants in the biopharmaceutical market.

text: Many of our 'small-molecule-drugs' are natural products from plants, or are synthetic compounds based on molecules found naturally in plants. However, the vast majority of the protein therapeutics (or biopharmaceuticals) we use are from animal or human sources, and are produced commercially in microbial or mammalian bioreactor systems. Over the last few years, it has become clear that plants have great potential for the production of human proteins and other protein-based therapeutic entities. Plants offer the prospect of inexpensive biopharmaceutical production without sacrificing product quality or safety, and following the success of several plant-derived technical proteins, the first therapeutic products are now approaching the market. In this review, the different plant-based production systems are discussed and the merits of transgenic plants are evaluated compared with other platforms. A detailed discussion is provided of the development issues that remain to be addressed before plants become an acceptable mainstream production technology. The many different proteins that have already been produced using plants are described, and a sketch of the current market and the activities of the key players is provided. Despite the currently unclear regulatory framework and general industry inertia, the benefits of plant-derived pharmaceuticals are now bringing the prospect of inexpensive veterinary and human medicines closer than ever before.


title: Translational readthrough-promoting drugs enhance pseudoknot-mediated suppression of the stop codon at the Moloney murine leukemia virus gag–pol junction.

text: Translational readthrough-promoting drugs enhance the incorporation of amino acids at stop codons and can thus bypass premature termination during protein synthesis. The polymerase (Pol) proteins of Moloney murine leukemia virus (MoMLV) are synthesized as a large Gag–Pol fusion protein, formed by the readthrough of a stop codon at the end of the gag ORF. The downstream pol ORF lacks its own start codon, and Pol protein synthesis is wholly dependent on translation of the upstream gag gene and the readthrough event for expression. Here, we explored the effects of readthrough-promoting drugs – aminoglycoside antibiotics and the small molecule ataluren – on the efficiency of readthrough of the stop codon in the context of the MoMLV genome. We showed that these compounds increased readthrough of the stop codon at the MoMLV gag–pol junction in vivo above the already high basal level and that the resulting elevated gag–pol readthrough had deleterious effects on virus replication. We also showed that readthrough efficiency could be driven to even higher levels in vitro, and that the combination of the small molecules and the RNA structure at the MoMLV stop codon could achieve extremely high readthrough efficiencies.


title: A sero-epizootiological study of porcine respiratory coronavirus in Belgian swine.

text: A porcine respiratory coronavirus (PRCV), antigenically closely related to transmissible gastroenteritis virus (TGEV), appeared in the European swine population in 1984. The present serological study was performed to obtain insight into the epizootiology of PRCV and of TGEV. PRCV-induced neutralizing antibodies were found in 90.6 per cent of the 160 sera collected from sows at slaughter, demonstrating the enzootic appearance of PRCV in the Belgian swine population. A serological study of fattening swine on 33 farms revealed that 11 farms situated in an area with a high farm density (all farms within 4 km2) and 11 on 22 closed breeding-fattening farms situated in areas with a low farm density (only one to four farms per 12 km2) were infected with PRCV throughout the year, whereas the other 11 closed breeding-fattening farms were temporarily free of PRCV. PRCV disappeared from the farms mainly in spring and summer. All the 11 farms became reinfected in autumn or winter, indicating that PRCV is regularly reintroduced in farms in the colder seasons. There was no correlation between the herd size and the temporary disappearance of PRCV from farms. It was observed on some farms that PRCV could infect pigs shortly after weaning in the presence of declining maternal antibodies, indicating that PRCV can persist on a farm by regularly infecting newly weaned pigs. TGEV-specific antibodies were found in 7.6 per cent of the 160 sera from the slaughterhouse sows. TGEV-specific antibodies were also detected in sera from fattening swine of 5 of the above mentioned 33 farms. TGEV-outbreaks were not observed on these farms.(ABSTRACT TRUNCATED AT 250 WORDS)


From the Centers for Disease Control and Prevention. Recommended childhood immunization schedule--United States, January 1995. Advisory committee on Immunization Practices, American Academy of Pediatrics, and American Academy of Family Physicians.


Lessons from SARS.


Conjunctiva-upper respiratory tract irrigation for early diagnosis of severe acute respiratory syndrome.


title: Robotic Minor Hepatectomy Offers a Favorable Learning Curve and May Result in Superior Perioperative Outcomes Compared with Laparoscopic Approach.

text: Minorhepatectomy (MH) is a common type of robotic-assisted liver resection, but few studies compared it with laparoscopic. We compared the perioperative outcomes of patients who underwent robotic (RH) or laparoscopic (LH) minor hepatectomy and evaluated the effect of surgeon's experience on outcomes. A prospective database was used to identify patients from 2009 through 2016 who underwent RH or LH. Two surgeons performed RH starting in 2014, whereas LH had been established before that. Of the 93 patients, 42 were in RH and 51 in LH group. The mean patient age, gender, race, American Society of Anesthesiologists score, proportion of patients with cirrhosis and hepatocellular carcinoma were similar. Operative time, estimated blood loss (EBL), conversion to open, 30-day complication rate, Clavien-Dindo grade ≥ 3 complications, and length of hospital stay (LOS) were similar. There was no difference in average tumor size, specimen volume, or achievement of R0 margin. In RH group, after completing 15 cases, there were no conversions to open. After 25 cases, EBL, LOS, and 30-day complication rate were improved as compared with LH. Perioperative outcomes of robotic MH are equivalent to laparoscopic. After approximately 25 cases, robotic-assisted MH may result in superior outcomes compared with laparoscopic.


title: New and emerging infectious diseases.

text: New emerging infectious diseases include "severe acute respiratory distress syndrome" (SARS) and avian influenza A H5N1. First cases of SARS, induced by a new strain of coronavirus, were described in China in 2002 and by May 2003 8360 cases and 764 deaths were reported by the World Health Organization. The disease can be transmitted from person to person and at the onset is characterized by nonspecific symptoms such as a fever of > 38 degrees C, dry cough, myalgia, and dyspnea. Adults can develop severe hypoxemia requiring intubation, whereas the course of the disease is generally benign in children. Avian influenza A H5N1 is another emerging infectious disease transmitted from avian species to humans, without clear evidence of transmission from human to human. The widespread outbreaks of H5N1 avian influenza in 2003-2004 have caused major problems for the poultry industry in many Asian countries. On January 2004 the disease crossed over to humans, for the first time in Vietnam, causing 74 deaths to date (mortality rate of 50%) in southeastern countries. Unlike SARS, the avian flu occurs in rural areas, where people live in intimate contact with birds, and many of the victims are children < 5 years of age. As for SARS, the World Health Organization has adopted a global action plan to control avian influenza among chickens and ducks and at the same time to limit the threat of a human flu pandemic.


title: [Analyses on one case of severe acute respiratory syndrome 'super transmitter' and chain of transmission].

text: OBJECTIVES To investigate the transmission process of severe acute respiratory syndrome (SARS) and to evaluate the infectiveness of SARS patients in different periods of disease epidemics. METHODS Standardized questionnaire was used to conduct case investigation and contact tracing by combining the field investigation and telephone interview. Transmission process, infectivity, transmission chain and contact history of SARS were studied through data analyses. RESULTS On 25th March 2003, a 91 year old man was admitted to Hospital J in Beijing with stroke and fever. He died on 30th March. From 31st March, there was an outbreak of SARS among his contacts in the family and in the hospital he was admitted to. Contacts would include his relatives, other co-patients and health care workers in the Hospital J. Chinese Field Epidemiology Training Program trainees conducted an investigation of the outbreak. Among the 207 contacts of the index cases through different generations, there were 36 cases of SARS (attack rate 17%) patients with one death. There were 12 cases having directly contact with the index case and 13 cases with one secondary case. The transmission chains of this outbreak could clearly be depicted. All the cases had close contacts during the symptomatic period of their index patients. Among the relatives, 85% of the cases had 3 - 5-day contact with their index patients after the onset of the illnesses. There was no significant difference between the two attack rates-70% for whose who had contact with the patient before and after illness onset) and 67% for those who only had contact after the onset of the illness. Out of the 44 social acquaintances and 38 of the family members who had contacts with the index patients during the incubation period, no one was found ill. Among the close contacts at the hospital who had no protection when providing care to the patient, the attack rate was found over 80%. CONCLUSIONS All the secondary cases of this outbreak had a history of direct and close contacts to the index patients after the onset of the illness. There was no evidence indicating that SARS cases were infectious during their incubation period.


title: [Determination of the frequency of human bocavirus and other respiratory viruses among 0-2 years age group children diagnosed as acute bronchiolitis].

text: Acute bronchiolitis, mostly seen in infants and younger children, is a lower respiratory tract infection frequently caused by viral agents. We aimed to determine the frequency of a broad panel of respiratory viruses including human bocavirus (HBoV) and to assess the clinical characteristics of acute bronchiolitis in a group of children under 24 months of age. A total of 62 children (45 male, 17 female; age range: 0-2 years) with the initial diagnosis of acute bronchiolitis and 33 healthy children (21 male, 12 female; age range: 0-2 years) as control group who were admitted to the Pediatrics Department of Mersin University Hospital, southern Turkey, from January to July 2010 were included in the study. Nasopharyngeal aspirates were collected from the study groups and the detection of respiratory viruses [respiratory syncytial virus (RSV) A & B; rhinovirus (RV); human metapneumovirus (hMPV) A & B; influenza virus type A [H1N1, H3N2, H1N1v], B & C; parainfluenza virus (PIV) type 1, 2, 3 & 4A/B; adenovirus (AdV); HBoV; coronavirus (CoV 229E); enterovirus (EV)], were performed by using a commercial system namely CLART®Pneumovir (Clinical Array Technology, Genomica, Spain) based on the principle of multiplex polymerase chain reaction (M-PCR) and DNA microarray. Demographic features, clinical and laboratory findings of the patients, treatment protocols and the relationship between the length of hospitalization and the viral agents determined were also evaluated. Of the 62 samples collected from bronchiolitis cases, at least one virus was detected in 52 (83.9%) and viral co-infections were detected in 31 (50%) of them. Including the co-infections, RSV was the most commonly identified virus (n= 21; 33.9%), followed by influenza A [H1N1] (n= 18; 29%), RV (n= 18; 29%), hMPV (n= 13; 21%), PIV (n= 10; 16.1%), AdV (n= 5; 8%), HBoV (n= 3; 4.8%) and EV (n= 1; 1.6%). Of the 33 samples from healthy children, at least one virus was detected in 21 (63.6%) and viral co-infections were detected in seven (21.2%) samples. Including the co-infections, the most commonly detected virus was RV (n= 10; 30.3%), followed by influenza A [H1N1] (n= 6; 18.1%), AdV (n= 6; 18.1%), RSV (n= 4; 12.1%) and PIV (n= 3; 9%), however HBoV and hMPV were not detected in the control group. The differences of demographic features (age, gender, history of atopy, exposure of smoking, length of breast-feeding, presence of school-age sibling) and frequency of virus detection (83.9% and 63.6%, respectively) between the patient and control groups were not statistically significant (p> 0.05). The most common complaints of patients on admission were cough (100%), runny nose (82.3%) and respiratory difficulty (71%), whereas fever was present in 21 (33.9%) patients. The most common findings on physical examination were prolonged expirium (98.4%), rhonchi (98.4%), rales (80.6%), tachypnea (71%) and tachycardia (67.7%). Pulmonary graphies revealed that diffuse air trappings were more common in virus-associated bronchiolitis (36/52; 69.2%) cases, on the other hand infiltrations were more common (6/10; 60%) in patients who were virus-negative (p< 0.05). The demographic features, clinical and laboratory findings, clinical severity scores, hospitalization rates and duration of hospitalizations in bronchiolitis cases did not show statistically significant differences between the viral agents (p> 0.05 for each parameter). However the rates of antibiotic and steroid use in hospitalized patients (24/34 and 5/34, respectively) were significantly higher than those of outpatients (7/28 and 0/28, respectively) (p= 0.001 and p= 0.03). Our data indicated a high rate (~84%) of respiratory viruses in children with bronchiolitis in the Mersin province and the detection of hMPV (21%) and HBoV (4.8%) only in the patient group encouraged their roles in the etiology of acute brochiolitis. It was concluded that viral etiology should be investigated in selected cases to prevent unnecessary antibiotic treatment and to initiate appropriate antiviral therapy when necessary.


title: Antigenic and plaque variations of serotype II feline infectious peritonitis coronaviruses.

text: Three feline coronavirus (FCoV) isolates KUK-H, M91-266, and M91-267 were examined to elucidate their biological and antigenic properties as well as disease potential in cats. Immune stainings of virus-infected cells by using FCoV type-specific monoclonal antibodies indicated that their antigenic specificity was serotype II. However, antigenic variations among these serotype II FCoVs were detected by neutralization assay with hyperimmune antisera against FCoVs and canine coronaviruses, and with experimentally infected cat sera; there were two subtypes in serotype II FCoVs. The isolates efficiently grew in fcwf-4 cell culture showing lytic CPE enough to form distinct plaques: when measured 48 hr after infection, plaque sizes of both M91-266 and M91-267 were approximately 1 mm in diameter, and a mixture of small (less than 1 mm in diameter) and large (approximately 3 mm in diameter) plaques were produced in the case of KUK-H. Strains KUK-H, M91-266 and M91-267 produced feline infectious peritonitis (FIP) in 50%, 67% and 89% of experimentally inoculated kittens, respectively. Furthermore, 80% of the kittens inoculated with the small plaque former of KUK-H developed FIP accompanied by more prominent clinical signs as well as pathological changes when compared with 28.6% of kittens inoculated with the large plaque former. These results suggest that serotype II FIPVs producing smaller size of plaques are more virulent than those producing larger size of plaques.


title: Coronavirus envelope assembly is sensitive to changes in the terminal regions of the viral M protein.

text: Recently we demonstrated that the co-expressed coronavirus membrane proteins have the capacity to assemble viral envelopes which are similar to normal virus particles in dimensions and appearance, and which can form independent of a nucleocapsid (Vennema et al., 1996). For the formation of these particles only the M and the E protein are required; the S protein is dispensable but is incorporated when present. As we illustrate here, this virus-like particle assembly system is an ideal tool to study the interactions between the essential assembly partners M and E in molecular detail. Taking a mutagenetic approach we demonstrate that envelope assembly is critically sensitive to changes in the primary structure of both terminal domains of the M protein. The effects were most dramatically observed after mutation of the carboxy-terminal domain where the deletion of just one single amino acid at the extreme terminus abolished particle formation almost completely. But also some subtle mutations in the amino-terminal domain were severely inhibitory to the assembly process. Interestingly, mutant M proteins that were themselves incompetent to support particle formation appeared to inhibit, in a concentration dependent manner, the assembly of particles by wild-type M and E protein.


title: Optimization of immunohistochemical methods using two different antigen retrieval methods on formalin-fixed paraffin-embedded tissues: experience with 63 markers.

text: Formalin fixation produces cross-links between the proteins and the fixative that alter the ability of some antibodies to recognize antigens. We used formalin-fixed, paraffin-embedded tissues to compare two different antigen retrieval methods for 63 antibodies used in the diagnosis of infectious and neoplastic diseases of animal species. Eighty-four percent of the antibodies needed some type of antigen retrieval for optimal results. Of those antibodies, 67.7% were monoclonal and 32.3% were polyclonal. Steam heat was the method of choice for 31 antibodies. Ten antibodies reacted only with steam heat, but 9 antibodies did not react when steam heat was used. Optimal results were obtained with enzyme digestion for 22 antibodies. Only 10 antibodies yielded optimal results without antigen retrieval; 64% of these antibodies were polyclonal. All antibodies against cytokeratins were optimally retrieved with proteinese K. Antigen retrieval appears to be necessary for the majority of antibodies when used with formalin-fixed, paraffin-embedded tissues.


title: Transumbilical single-incision total laparoscopic hysterectomy: technique and initial experience in Turkey.

text: OBJECTIVES The aim of the study is to determine the results of our initial experience of 32 cases who underwent single incision total laparoscopic hysterectomy MATERIALS AND METHODS Thirty-two patients who underwent transumbilical single incision total laparoscopic hysterectomy between March 2009 and February 2011 were reviewed retrospectively at the Department of Obstetrics and Gynecology Ege University Faculty of Medicine, Izmir, Turkey Articulating and rigid instruments, 30 degrees 10 mm telescope, SILS port and advanced bipolar and mechanical energy modalities were used during the procedure. Duration of surgery length of hospital stay mean blood loss and postoperative complications were assessed. RESULTS Mean age of patients and mean operation time were 48 years (ranged 42-55) and 108 minutes (ranged 80-180), respectively. In all cases vaginal cuffs were closed with either intracorporeal or extracorporeal separate sutures. There were no intraoperative complications. All patients were discharged within 48 hours postoperatively The mean length of hospital stay and mean blood loss were as 1.6 days (range 1-2) and 45 ml (range 30-100), respectively CONCLUSIONS Total laparoscopic hysterectomy performed through transumbilical single incision is technically feasible and safe. Development of advanced flexible instrumentation and visualization platform may facilitate this new operative approach.


title: A review of the research literature on evidence-based healthcare design.

text: OBJECTIVE This report surveys and evaluates the scientific research on evidence-based healthcare design and extracts its implications for designing better and safer hospitals. BACKGROUND It builds on a literature review conducted by researchers in 2004. METHODS Research teams conducted a new and more exhaustive search for rigorous empirical studies that link the design of hospital physical environments with healthcare outcomes. The review followed a two-step process, including an extensive search for existing literature and a screening of each identified study for the relevance and quality of evidence. RESULTS This review found a growing body of rigorous studies to guide healthcare design, especially with respect to reducing the frequency of hospital-acquired infections. Results are organized according to three general types of outcomes: patient safety, other patient outcomes, and staff outcomes. The findings further support the importance of improving outcomes for a range of design characteristics or interventions, including single-bed rooms rather than multibed rooms, effective ventilation systems, a good acoustic environment, nature distractions and daylight, appropriate lighting, better ergonomic design, acuity-adaptable rooms, and improved floor layouts and work settings. Directions for future research are also identified. CONCLUSIONS The state of knowledge of evidence-based healthcare design has grown rapidly in recent years. The evidence indicates that well-designed physical settings play an important role in making hospitals safer and more healing for patients, and better places for staff to work.


docid: szjsouyr

title: Helper-dependent adenoviral vectors in experimental gene therapy.

text: In the majority of potential applications gene therapy will require an effective transfer of a transgene in vivo resulting in high-level and long-term transgene expression, all in the absence of significant toxicity or inflammatory responses. The most efficient vehicles for delivery of foreign genes to the target tissues are modified adenoviruses. Adenoviral vectors of the first generation, despite the high infection efficacy, have an essential drawback: they induce strong immune response, which leads to short term expression of the transgene, and limits their usefulness in clinical trials. In contrast, helper-dependent adenoviral vectors (HdAd) lacking all viral coding sequences display only minimal immunogenicity and negligible side-effects, allowing for long-term transgene expression. Thus, HdAd vehicles have become the carrier of choice for adenoviral vector-mediated experimental gene therapy, effectively used in animal models for delivery of transgenes into the liver, skeletal muscle, myocardium or brain. Strong and long-lasting expression of therapeutic genes has allowed for successful treatment of dyslipidemias, muscular dystrophy, obesity, hemophilia, and diabetes. Additionally, the large cloning capacity of HdAd, up to 37 kb, facilitates the use of physiologically regulated, endogenous promoters, instead of artificial viral promoter sequences. This enables also generation of the single vectors expressing multiple genes, which can be potentially useful for treatment of polygenic diseases. In this review we characterize the basic features of HdAd vectors and describe some of their experimental and potential clinical applications.


Unjustified restrictions on letters to the editor.


Induction of an immune response to transmissible gastroenteritis coronavirus using vectors with enteric tropism.


Defective interfering particles of coronavirus.


docid: pveldpqy

title: Severe acute respiratory syndrome and the innate immune responses: modulation of effector cell function without productive infection.

text: Severe acute respiratory syndrome (SARS) caused by a novel human coronavirus (CoV), designated SARS-CoV, is a highly contagious respiratory disease with the lungs as a major target. Although the exact mechanism of SARS-CoV pathogenesis remains unknown, an intense, ill-regulated local inflammatory response has been suggested as partially responsible for the devastating lung pathology. We investigated the interaction of SARS-CoV with human macrophages (Mphi) and dendritic cells (DC), two key innate immune cells of the host immune system, by focusing on their susceptibility to viral infection and subsequent responses (e.g., phenotypic maturation, T cell-priming activity, phagocytosis, and cytokine production). We found neither cell to be permissive for SARS-CoV replication. However, incubation of Mphi and DC with live, but not gamma irradiation-inactivated, viruses appeared to better sustain their viability. Also, exposure to infectious SARS-CoV led to the phenotypic and functional maturation of DC, with regard to MHC class II and costimulatory molecule expression, T cell-stimulatory capacity, and cytokine production, respectively. Cytokine production was also observed for Mphi, which were refractory to cell surface phenotypic changes. Strikingly, live SARS-CoV could further prime cell types to respond to a suboptimal dose of bacterial LPS (100 ng/ml), resulting in massive release of IL-6 and IL-12. However, the endocytic capacity (e.g., Ag capture) of Mphi was significantly compromised upon exposure to infectious SARS-CoV. This study is the first demonstration that although SARS-CoV does not productively infect human Mphi or DC, it appears to exert differential effects on Mphi and DC maturation and functions, which might contribute to SARS pathogenesis.


title: An interactive Internet-based system for tracking upper limb motion in home-based rehabilitation.

text: In this paper, we introduce an interactive telecommunication system that supports video/audio signal acquisition, data processing, transmission, and 3D animation for post stroke rehabilitation. It is designed for stroke patients to use in their homes. It records motion exercise data, and immediately transfers this data to hospitals via the internet. A real-time videoconferencing interface is adopted for patients to observe therapy instructions from therapists. The system uses a peer-to-peer network architecture, without the need for a server. This is a potentially effective approach to reducing costs, allowing easy setup and permitting group-rehabilitation sessions. We evaluate this system using the following steps: (1) motion detection in different movement patterns, such as reach, drink, and reach-flexion; (2) online bidirectional visual telecommunication; and (3) 3D rendering using a proposed offline animation package. This evaluation has subjectively been proved to be optimal.


title: Health evaluation of free-ranging Humboldt penguins (Spheniscus humboldti) in Peru.

text: As part of ongoing ecological studies of Humboldt penguins (Spheniscus humboldti) at Punta San Juan, Ica Department, Peru, health surveys were conducted in November 1992, 1993, and 1994. In the three surveys, 98 birds in total were handled for examination, and blood was collected for laboratory analysis from 90 of these birds. All birds seemed to be in good condition. Body weights of females were significantly lower in 1994 than in the other years. Fleas (Parapsyllus humboldti) and ticks (Ornithodoros amblus) were found on the penguins and in their nests. Females had significantly higher plasma calcium and phosphorus levels, and they had lower weights than males. No other differences were found between the sexes. Hematology, plasma chemistries, and plasma mineral levels varied between years. Positive antibody titers for Chlamydophila psittaci (62%), avian adenovirus (7%; 1994 only), paramyxovirus-2 (7%; 1993 only), and Salmonella Pullorum (7%) were found. Plasma chemistry and mineral levels differed between individuals testing positive vs. negative on serologic tests for avian adenovirus and Salmonella Pullorum. Serologic tests for antibodies to avian influenza A virus, avian encephalomyelitis virus, infectious bronchitis virus, avian reovirus, duck viral enteritis virus, equine encephalitis (eastern, western, and Venezuelan) viruses, infectious bursal disease virus, infectious laryngotracheitis virus, Aspergillus sp., and paramyxovirus-1 and -3 were negative. All chlorinated pesticide and polychlorinated biphenyl analyses were below detectable limits.


title: [Dynamic changes of immunoglobulin G in convalescents who have suffered from severe acute respiratory syndrome patients].

text: OBJECTIVE To investigate the dynamic changes of the antibody specific to severe acute respiratory syndrome (SARS) coronavirus in convalescents who have suffered from SARS. METHODS Samples of peripheral blood were collected twice during the first 2 weeks after discharge and then once every 2 - 4 weeks from 310 convalescents of SARS, 131 males and 179 females, aged 18 - 74, discharged from hospitals in Beijing April 3 to June 20 2003 with the average discharge date of June 10, to detect the level of immunoglobulin G (IgG) by ELISA. A curve of level of IgG was drawn to describe the change by the specific month in the year. RESULTS Most of the 310 convalescents underwent successive or non-successive testing for 2 - 4 months and 15 were tested for 5 months, 15 for 6 months, and 2 for 7 months. IgG was detected in each sample with the mean of A value of 0.97 +/- 0.37 (0.197 - 1.849). The mean IgG level peaked in July (1.203), about 35 days after discharge, and then gradually declined to 0.857 in December, a decline by 27.3%. CONCLUSION All SARS patients generate specific antibody against the coronavirus. However, the antibody level gradually decline with the lapse of time during the convalescence. Long-term surveillance of the change of antibody is necessary.


title: Investigation of immunogenic T-cell epitopes in SARS virus nucleocapsid protein and their role in the prevention and treatment of SARS infection.

text: 1. A novel HLA-A2.1-specific SARS coronavirus (SARS-CoV) nucleocapsid (N) protein epitope (N220-N228 LALLLLDRL) able to activate cytotoxic T cells in vitro has been identified. 2. When used with a single-chain-trimer system, the SARS-CoV N protein epitope (N220-N228 LALLLLDRL) can stimulate a cytotoxic T-cell response against N-protein expressing cells in the HLA-A2.1K(b) transgenic mouse model.


title: Integrative genomic, transcriptional, and proteomic diversity in natural isolates of the human pathogen Burkholderia pseudomallei.

text: Natural isolates of pathogenic bacteria can exhibit a broad range of phenotypic traits. To investigate the molecular mechanisms contributing to such phenotypic variability, we compared the genomes, transcriptomes, and proteomes of two natural isolates of the gram-negative bacterium Burkholderia pseudomallei, the causative agent of the human disease melioidosis. Significant intrinsic genomic, transcriptional, and proteomic variations were observed between the two strains involving genes of diverse functions. We identified 16 strain-specific regions in the B. pseudomallei K96243 reference genome, and for eight regions their differential presence could be ascribed to either DNA acquisition or loss. A remarkable 43% of the transcriptional differences between the strains could be attributed to genes that were differentially present between K96243 and Bp15682, demonstrating the importance of lateral gene transfer or gene loss events in contributing to pathogen diversity at the gene expression level. Proteins expressed in a strain-specific manner were similarly correlated at the gene expression level, but up to 38% of the global proteomic variation between strains comprised proteins expressed in both strains but associated with strain-specific protein isoforms. Collectively, >65 hypothetical genes were transcriptionally or proteomically expressed, supporting their bona fide biological presence. Our results provide, for the first time, an integrated framework for classifying the repertoire of natural variations existing at distinct molecular levels for an important human pathogen.


title: SARS preventive and risk behaviours of Hong Kong air travellers.

text: This study aims to investigate Severe Acute Respiratory Syndrome (SARS)-related behaviours of travellers returning to Hong Kong by air. A total of 820 travellers returning to Hong Kong by air were interviewed about their SARS-related behaviours in April 2003. Three quarters of the respondents wore a mask most/all of the time on board, 15% did so in public places at the travel destination. Perceived susceptibility to SARS at the destination predicted mask-wearing in public places and avoidance of crowded places, and perceived efficacy was a predictor for mask-wearing during flight. Approximately 16% of the respondents stated that they would delay their medical consultation for flu-like symptoms until returning to Hong Kong. Nearly 18.2% stated that they would not wear a mask in public places at the destination if they had flu-like symptoms. Education programmes, special services and effective thermal screening are required to minimize the chance of the spread of SARS by air travellers.


title: Pleiotropic costs of niche expansion in the RNA bacteriophage phi 6.

text: Natural and experimental systems have failed to universally demonstrate a trade-off between generalism and specialism. When a trade-off does occur it is difficult to attribute its cause to antagonistic pleiotropy without dissecting the genetic basis of adaptation, and few previous experiments provide these genetic data. Here we investigate the evolution of expanded host range (generalism) in the RNA virus phi6, an experimental model system allowing adaptive mutations to be readily identified. We isolated 10 spontaneous host range mutants on each of three novel Pseudomonas hosts and determined whether these mutations imposed fitness costs on the standard laboratory host. Sequencing revealed that each mutant had one of nine nonsynonymous mutations in the phi6 gene P3, important in host attachment. Seven of these nine mutations were costly on the original host, confirming the existence of antagonistic pleiotropy. In addition to this genetically imposed cost, we identified an epigenetic cost of generalism that occurs when phage transition between host types. Our results confirm the existence in phi6 of two costs of generalism, genetic and environmental, but they also indicate that the cost is not always large. The possibility for cost-free niche expansion implies that varied ecological conditions may favor host shifts in RNA viruses.


title: Genetic diversity of avian infectious bronchitis coronavirus in recent years in China.

text: Fifty-six isolates of avian infectious bronchitis virus (IBV) were obtained from different field outbreaks in China in 2010, and they were genotyped by comparison with 19 reference strains in the present study. The results showed that LX4-type isolates are still the predominant IBVs circulating in chicken flocks in China, and these isolates could be grouped further into two clusters. Viruses in each cluster had favored amino acid residues at different positions in the S1 subunit of the spike protein. In addition, a recombination event was observed to have occurred between LX4- and tl/CH/LDT3/03I-type strains, which contributed to the emergence of a new strain. The most important finding of the study is the isolation and identification of Taiwan II-type (TW II-type) strains of IBV in mainland China in recent years. The genome of TW II-type IBV strains isolated in mainland China has experienced mutations and deletions, as demonstrated by comparison of the entire genome sequence with those of IBV strains isolated in Taiwan. Pathogenicity testing and sequence analysis of the 3' terminal untranslated region revealed that TW II-type IBV strains isolated in mainland China have a close relationship with the embryo-passaged, attenuated TW2296/95.


title: Effect of tracheostomy tube on work of breathing: Comparison of pre- and post-decannulation.

text: OBJECTIVE To describe and compare the work of breathing (WOB) during spontaneous breathing under four conditions: (1) breathing through a tracheostomy tube with an inflated cuff, (2) breathing through the upper airway (UA) with a deflated cuff and occluded tube, (3) breathing through the UA with an occluded cuffless tube, and (4) postdecannulation. PATIENTS AND METHODS Patients who tolerated an occluded cuffless tube were included. Ventilatory variables and esophageal pressure were recorded. The pressure-time product (PTP), PTP/min, and PTP/min/tidal volume (PTP/min/VT) were measured. Each condition was measured for 5 min with a 15 min time interval between evaluations. Quantitative data are expressed as mean ± standard deviation. Single-factor analysis of variance was used, and the Games-Howell test was used for post hoc analysis of comparisons between group means (P ≤ 0.05). RESULTS Eight patients were studied under each of the four conditions described above. Statistically significant differences were found for PTP, PTP/min, and PTP/min/VT. In the post hoc analysis for PTP, significant differences among all conditions were found. For PTP/min, there was no significant difference between Conditions 2 and 4 (P = 0.138), and for PTP/min/VT, there was no significant difference between Conditions 1 and 2 (P = 0.072) or between Conditions 2 and 3 (P = 0.106). A trend toward a higher PTP, PTP/min, and PTP/min/VT was observed when breathing through a cuffless tracheostomy tube. CONCLUSION The four conditions differed with respect to WOB. Cuff inflation could result in a reduced WOB because there is less dead space. Cuffless tracheostomy tubes generate increased WOB, perhaps due to the material deformity caused by body temperature.


title: A test for constant fatality rate of an emerging epidemic: with applications to severe acute respiratory syndrome in Hong Kong and Beijing.

text: The etiology, pathogenesis, and prognosis for a newly emerging disease are generally unknown to clinicians. Effective interventions and treatments at the earliest possible times are warranted to suppress the fatality of the disease to a minimum, and inappropriate treatments should be abolished. In this situation, the ability to extract most information out of the data available is critical so that important decisions can be made. Ineffectiveness of the treatment can be reflected by a constant fatality over time while effective treatment normally leads to a decreasing fatality rate. A statistical test for constant fatality over time is proposed in this article. The proposed statistic is shown to converge to a Brownian motion asymptotically under the null hypothesis. With the special features of the Brownian motion, we are able to analyze the first passage time distribution based on a sequential tests approach. This allows the null hypothesis of constant fatality rate to be rejected at the earliest possible time when adequate statistical evidence accumulates. Simulation studies show that the performance of the proposed test is good and it is extremely sensitive in picking up decreasing fatality rate. The proposed test is applied to the severe acute respiratory syndrome data in Hong Kong and Beijing.


docid: rymidhit

title: Impact of respiratory virus infection in patients with chronic chest disease.

text: This study investigated the morbidity associated with respiratory virus infections in patients with well-documented chest disease, and the risk of transmission between close contacts. Patients informed the study team if they were exposed to a family member or colleague with a cold. Patients and symptomatic index cases recorded respiratory symptoms during the study period. Acute nasopharyngeal swabs and paired sera were obtained for viral diagnosis. Twenty-five (43%) of 58 recorded exposures resulted in a symptomatic illness and 16 (28%) patients developed lower respiratory tract symptoms. Sixteen (64%) of the 25 symptomatic patients contacted their general practitioner, 14 (56%) received antibiotics and 4 (16%) were hospitalized. Mean duration of illness was 10.6 days in symptomatic patients and 5.7 days in their corresponding index cases (P < 0.005). Mean symptom scores were 100.6 in symptomatic patients and 62.2 in index cases (P < 0.01). Respiratory viruses were identified in 19 (33%) episodes. Rhinovirus, coronavirus and respiratory syncytial virus infections were all associated with lower respiratory tract exacerbations. Respiratory tract symptoms following exposure to a cold were comparatively severe in these patients with chronic chest disease. This group of patients might gain particular benefit from the introduction of effective vaccines or antiviral therapy.


title: Characterization of an essential RNA secondary structure in the 3' untranslated region of the murine coronavirus genome.

text: We have previously identified a functionally essential bulged stem-loop in the 3' untranslated region of the positive-stranded RNA genome of mouse hepatitis virus. This 68-nucleotide structure is composed of six stem segments interrupted by five bulges, and its structure, but not its primary sequence, is entirely conserved in the related bovine coronavirus. The functional importance of individual stem segments of this stem-loop was characterized by genetic analysis using targeted RNA recombination. We also examined the effects of stem segment mutations on the replication of mouse hepatitis virus defective interfering RNAs. These studies were complemented by enzymatic and chemical probing of the stem-loop. Taken together, our results confirmed most of the previously proposed structure, but they revealed that the terminal loop and an internal loop are larger than originally thought. Three of the stem segments were found to be essential for viral replication. Further, our results suggest that the stem segment at the base of the stem-loop is an alternative base-pairing structure for part of a downstream, and partially overlapping, RNA pseudoknot that has recently been shown to be necessary for bovine coronavirus replication.


title: Murine coronavirus nonstructural protein ns2 is not essential for virus replication in transformed cells.

text: Two isolates of the murine hepatitis virus (MHV) strain JHM, which differed in their ability to express the nonstructural gene product ns2, were characterized. The MHV Wb3 isolate encodes a 30,000-molecular-weight ns2 protein that can be readily detected in infected cells by using a specific monoclonal antibody, MAb 2A. The MHV Wb1 isolate is a deletion mutant that lacks a functional ns2 gene and the transcriptional signals required for the synthesis of an ns2 mRNA. However, there are no obviously significant differences in the growth of the MHV Wb1 and MHV Wb3 isolates in continuous cell lines or in the synthesis of viral mRNAs or proteins in infected cells. These results demonstrate that the ns2 gene product is not essential for MHV replication in transformed murine cells and suggests that the function of the ns2 gene may only be manifest in vivo.


docid: qgyokrsr

title: Electrocardiographic changes following rabbit coronavirus-induced myocarditis and dilated cardiomyopathy.

text: Rabbit Coronavirus (RbCV) infection was divided into two phases based upon day of death and pathologic findings. During the acute phase (days 2-5) heart weights (HW) and heart weight-to-body weight (HW/BW) ratios were increased with striking dilation of the right ventricle. These changes as well as increased dilation of the left ventricle were especially pronounced during the subacute phase (days 6-12). Myocytolysis, pulmonary edema, and degeneration and necrosis of myocytes, were seen during both phases. Myocarditis, pleural effusion, calcification of myocytes, and congestion in the liver and lungs were seen in the subacute phase. Electrocardiograms (ECGs) exhibited low voltage, nonspecific ST-T wave changes, sinus tachycardia, occasional ventricular and supraventricular premature complexes and 2(0) AV block consistent with myocarditis and heart failure. Forty-one percent of the survivors exhibited increased HW and HW/BW ratios, biventricular dilation, interstitial and replacement fibrosis, myocyte hypertrophy and myocarditis. ECGs exhibited nonspecific ST-T wave changes, sinus arrhythmia, occasional ventricular and supraventricular premature complexes and 2(0) AV block. These data suggest that RbCV infection may result in viral myocarditis and heart failure with a proportion of survivors progressing into DCM.


title: Splenic Bed Laparoscopic Splenectomy Approach for Massive Splenomegaly Secondary to Portal Hypertension and Liver Cirrhosis.

text: This study was performed to evaluate the feasibility of the splenic bed laparoscopic splenectomy approach (SBLS) for massive splenomegaly (≥30 cm) in patients with hypersplenism secondary to portal hypertension and liver cirrhosis. Patients who underwent laparoscopic splenectomy (LS) from January 2012 to December 2016 were retrospectively reviewed. We performed LS in 83 patients with massive splenomegaly (≥30 cm) secondary to portal hypertension and liver cirrhosis. Of these patients, 37 underwent the SBLS and 46 underwent anterior LS (ALS). Five patients in the ALS group and none in the SBLS group underwent conversion to open surgery. The operation time, intraoperative blood loss volume, transfusion volume, frequency of transfusion, hemorrhage of short gastric vessels, conversion rate, postoperative hospital stay, and incidence of pancreatic fistula were all significantly lower in the SBLS than ALS group (all P < 0.05). No death or postoperative bleeding occurred in the two groups, and there were no significant differences in age, gender, spleen size, hemoglobin level, platelet count, prothrombin time, Child-Pugh class, hypoproteinemia, or ascites (all P > 0.05). The SBLS is more feasible and effective than ALS in patients with massive splenomegaly (≥30 cm) secondary to portal hypertension and liver cirrhosis.


title: [Healthcare, European Stability Mechanism and public funding following the Covid-19 pandemic].

text: The aim of this editorial is to illustrate the new public funding framework of the Italian National Health System following the Covid-19 pandemic. The document reviews the measures put in place by the Italian Government and European Institutions such as the European Commission (EC), the European Central Bank (ECB) and the European Stability Mechanism (ESM) to deal with this health crisis and subsequent severe economic recession, with particular reference to sources and uses of resources. The use of new budgetary financial spaces in deficit entails greater attention to the assessment of interventions and makes it necessary to keep expenditure under strict control. At the same time, the remodeling of expenditure within its aggregates, public investment in innovation, and the removal of administrative obstacles can strengthen the capacity of the healthcare system to meet the extraordinary needs deriving from the spread of Covid-19 and its resilience to future health shocks.


title: [Clinical observation of 6 severe COVID-19 patients treated with plasma exchange or tocilizumab].

text: OBJECTIVE To observe the clinical effect of plasma exchange and tocilizumab in treatment of patients with severe coronavirus disease 2019 (COVID-19). METHODS Six patients with severe COVID-19 admitted in First Affiliated Hospital of Bengbu Medical College from January 25 to February 25, 2020. Three patients were treated with plasma exchange and three patients were treated with tocilizumab. The effect on excessive inflammatory reaction of plasma exchange and tocilizumab was observed. RESULTS The C-reactive protein (CRP) and IL-6 levels were significantly decreased and the lymphocyte and prothrombin time were improved in 3 patients after treatment with plasma exchange; while inflammation level was not significantly decreased, and lymphocyte and prothrombin time did not improve in 3 patients treated with tocilizumab. CONCLUSIONS For severe COVID-19 patients with strong inflammatory reaction, plasma exchange may be preferred.


title: Detection of beet yellows closterovirus methyltransferase-like and helicase-like proteins in vivo using monoclonal antibodies.

text: In the positive-stranded RNA genome of beet yellows closterovirus (BYV), the 5'-terminal ORF 1a encodes a 295 kDa polyprotein with the domains of papain-like cysteine proteinase, methyltransferase (MT) and helicase (HEL), whereas ORF 1b encodes an RNA-dependent RNA polymerase. Eleven and five hybridoma cell lines secreting monoclonal antibodies (MAbs) were derived from mice injected with the bacterially expressed fragments of the BYV 1a product encompassing the MT and HEL domains, respectively. On immunoblots of protein from BYV-infected Tetragonia expansa plants, four MAbs against the MT recognized a approximately 63 kDa protein, and two MAbs against the HEL recognized a approximately 100 kDa protein. Both the methyltransferase-like protein and the helicase-like protein were found mainly in the fractions of large organelles (P1) and membranes (P30) of the infected plants. These data clearly indicate that (i) the BYV methyltransferase-like and helicase-like proteins, like other related viral enzymes, are associated with membrane compartments in cells, and (ii) the 1a protein, apart from the cleavage by the leader papain-like proteinase that is expected to produce the 66 kDa and 229 kDa fragments, undergoes additional processing by a virus-encoded or cellular proteinase.