title: Isolation of high-affinity single-chain antibodies against Mycobacterium avium subsp. paratuberculosis surface proteins from sheep with Johne's disease.

text: Johne's disease, caused by infection with Mycobacterium avium subsp. paratuberculosis, causes significant economic losses to the livestock farming industry. Improved investigative and diagnostic tools-necessary to understand disease processes and to identify subclinical infection-are much sought after. Here, we describe the production of single-chain antibodies with defined specificity for M. avium subsp. paratuberculosis surface proteins. Single-chain antibodies (scFv) were generated from sheep with Johne's disease by cloning heavy-chain and lambda light-chain variable regions and expressing these in fusion with gene III of filamentous phages. Two scFv clones (designated SurfS1.2 and SurfS2.2) were shown to be immunoreactive against M. avium subsp. paratuberculosis surface targets by flow cytometry, and immunoblotting identified specificity for a 34-kDa proteinase-susceptible determinant. Both antibodies were cross-reactive against Mycobacterium avium subsp. avium but nonreactive against Mycobacterium bovis or Mycobacterium phlei cells and were shown to be capable of enriching M. avium subsp. paratuberculosis cells by a factor of approximately 10(6)-fold when employed in magnetic bead separation of mixed Mycobacterium sp. cultures. Further, magnetic bead separation using SurfS1.2 and SurfS2.2 was capable of isolating as few as 10(3) M. avium subsp. paratuberculosis cells from ovine fecal samples, indicating the diagnostic potential of these reagents. Finally, inclusion of SurfS1.2 or SurfS2.2 in in vitro broth culture with M. avium subsp. paratuberculosis indicated that surface binding activity did not impede bacterial growth, although colony clumping was prevented. These results are discussed in terms of the potential use of single-chain phage display monoclonal antibodies as novel diagnostic reagents.


title: T-cell immune responses in the brain and their relevance for cerebral malignancies.

text: In order that cellular immune responses afford protection without risk to sensitive normal tissue, they must be adapted to individual tissues of the body. Nowhere is this more critical than for the brain, where various passive and active mechanisms maintain a state of immune privilege that can limit high magnitude immune responses. Nevertheless, it is now clear that immune responses are induced to antigens in the brain, including those expressed by cerebral malignancies. We discuss hypotheses of how this can occur, although details such as which antigen presenting cells are involved remain to be clarified. Antitumor responses induced spontaneously are insufficient to eradicate malignant astrocytomas; many studies suggest that this can be explained by a combination of low level immune response induction and tumor mediated immunosuppression. A clinical objective currently pursued is to use immunotherapy to ameliorate antitumour immunity. This will necessitate a high level immune response to ensure sufficient effector cells reach the tumor bed, focused cytotoxicity to eradicate malignant cells with little collateral damage to critical normal cells, and minimal inflammation. To achieve these aims, priority should be given to identifying more target antigens in astrocytoma and defining those cells present in the brain parenchyma that are essential to maintain antitumour effector function without exacerbating inflammation. If we are armed with better understanding of immune interactions with brain tumor cells, we can realistically envisage that immunotherapy will one day offer hope to patients with currently untreatable neoplastic diseases of the CNS.


Interim schedule for pregnant women and children during the COVID-19 pandemic.


title: Performance of single-step gel-based reverse transcription-PCR (RT-PCR) assays equivalent to that of real-time RT-PCR assays for detection of the severe acute respiratory syndrome-associated coronavirus.

text: Simple gel-based one-step reverse transcription-PCR (RT-PCR) assays, used to investigate patients during the 2003 severe acute respiratory syndrome (SARS) outbreak in Singapore, were found to be as sensitive as commercial and in-house real-time RT-PCR assays. The detection limit was approximately 1 genome equivalent (GE) per 5 microl PCR mixture. One PFU of SARS coronavirus was estimated to be 258 +/- 46 GE.


title: Infections, connective tissue diseases and vasculitis.

text: In genetically predisposed individuals, viruses, bacteria, or parasitic infectious agents are suspected of inducing autoimmunity and/or exacerbating autoimmune rheumatic diseases (ARD) once self-tolerance is broken. Although direct evidence for this association is still lacking, numerous data from animal models as well as from humans support the hypothesis of a direct contribution of pathogens to the induction of several ARD. This review focuses on the possible role of infectious agents as triggers of autoimmunity in systemic lupus erythematosus, polymyositis-dermatomyositis, antiphospholipid antibody syndrome, and primary vasculitis. Indeed, vasculitis may be a clinical manifestation of an infectious disease (secondary vasculitis). In addition, immune response abnormalities and immunosuppressive medications may be responsible for the high percentage of infectious complications in ARD patients. Recent therapeutic approaches aimed at lowering doses of cytotoxic agents and shortening duration of treatment with the most toxic drugs, have proved to be as effective as conventional regimens. New drugs and strategies aimed at preventing infections could further improve the outcome of ARD patients.


title: Baculovirus stimulates antiviral effects in mammalian cells.

text: Herein, we report that Autographa californica nucleopolyhedrovirus, a member of the Baculoviridae family, is capable of stimulating antiviral activity in mammalian cells. Baculoviruses are not pathogenic to mammalian cells. Nevertheless, live baculovirus is shown here to induce interferons (IFN) from murine and human cell lines and induces in vivo protection of mice from encephalomyocarditis virus infection. Monoclonal antibodies specific for the baculovirus envelope gp67 neutralize baculovirus-dependent IFN production. Moreover, UV treatment of baculovirus eliminates both infectivity and IFN-inducing activity. In contrast, the IFN-inducing activity of the baculovirus was unaffected by DNase or RNase treatment. These data demonstrate that IFN production can be induced in mammalian cells by baculovirus even though the cells fail to serve as a natural host for an active viral infection. Baculoviruses, therefore, provide a novel model in which to study at least one alternative mechanism for IFN induction in mammalian cells.


title: Full-length open reading frame of a recombinant hepatitis C virus strain from St Petersburg: proposed mechanism for its formation.

text: The full-length ORFs for the hepatitis C virus recombinant RF1_2k/1b (N687) and the non-recombinant 1b strain N589 were sequenced. A single recombination point was found and the sizes of the genes (C, E1, E2, p7, NS2, NS3, NS4 and NS5) were according to the parental subtypes. The PKR-eIF2alpha phosphorylation site homology domain sequence of the E2 protein was identical to those of genotype 2 strains, while the IFN-alpha-sensitivity-determining region of the NS5A protein was identical to those of interferon-resistant 1b strains. For the parental strains, two hairpin structures, HS1 and HS2, were predicted for the plus-strand up- and downstream of the crossover site, which were not present in the recombinant strain. HS2 shared similarity with the motif1 hairpin of turnip crinkle virus RNA that binds to the RNA-dependent RNA polymerase and facilitates 3'-terminal extension during recombination. This study suggests that RF1_2k/1b has emerged by homologous recombination during minus-strand synthesis via template switching because of constraints imposed by the HS1 hairpin of the 3'-parental genome.


title: Assembly of severe acute respiratory syndrome coronavirus RNA packaging signal into virus-like particles is nucleocapsid dependent.

text: The severe acute respiratory syndrome coronavirus (SARS-CoV) was recently identified as the etiology of SARS. The virus particle consists of four structural proteins: spike (S), small envelope (E), membrane (M), and nucleocapsid (N). Recognition of a specific sequence, termed the packaging signal (PS), by a virus N protein is often the first step in the assembly of viral RNA, but the molecular mechanisms involved in the assembly of SARS-CoV RNA are not clear. In this study, Vero E6 cells were cotransfected with plasmids encoding the four structural proteins of SARS-CoV. This generated virus-like particles (VLPs) of SARS-CoV that can be partially purified on a discontinuous sucrose gradient from the culture medium. The VLPs bearing all four of the structural proteins have a density of about 1.132 g/cm(3). Western blot analysis of the culture medium from transfection experiments revealed that both E and M expressed alone could be released in sedimentable particles and that E and M proteins are likely to form VLPs when they are coexpressed. To examine the assembly of the viral genomic RNA, a plasmid representing the GFP-PS580 cDNA fragment encompassing the viral genomic RNA from nucleotides 19715 to 20294 inserted into the 3' noncoding region of the green fluorescent protein (GFP) gene was constructed and applied to the cotransfection experiments with the four structural proteins. The SARS-CoV VLPs thus produced were designated VLP(GFP-PS580). Expression of GFP was detected in Vero E6 cells infected with the VLP(GFP-PS580), indicating that GFP-PS580 RNA can be assembled into the VLPs. Nevertheless, when Vero E6 cells were infected with VLPs produced in the absence of the viral N protein, no green fluorescence was visualized. These results indicate that N protein has an essential role in the packaging of SARS-CoV RNA. A filter binding assay and competition analysis further demonstrated that the N-terminal and C-terminal regions of the SARS-CoV N protein each contain a binding activity specific to the viral RNA. Deletions that presumably disrupt the structure of the N-terminal domain diminished its RNA-binding activity. The GFP-PS-containing SARS-CoV VLPs are powerful tools for investigating the tissue tropism and pathogenesis of SARS-CoV.


title: A nested set of eight RNAs is formed in macrophages infected with lactate dehydrogenase-elevating virus.

text: Total RNA was extracted from primary cultures of mouse macrophages isolated from 10-day-old mice 6 to 12 h postinfection with lactate dehydrogenase-elevating virus (LDV). Poly(A)+ RNA was extracted from spleens of 18-h LDV-infected mice. The RNAs were analyzed by Northern (RNA) blot hybridization with a number of LDV-specific cDNAs as probes. A cDNA representing the nucleocapsid protein (VP-1) gene located at the 3' terminus of the viral genome (E. K. Godeny, D. W. Speicher, and M. A. Brinton, Virology 177:768-771, 1990) hybridized to viral genomic RNA of about 13 kb plus seven subgenomic RNAs ranging in size from about 1 to about 3.6 kb. Two other cDNA clones hybridized only to the four or five largest subgenomic RNAs, respectively. In contrast, two cDNAs encoding continuous open reading frames with replicase and zinc finger motifs hybridized only to the genomic RNA. The replicase motif exhibited 75% amino acid identity to that of the 1b protein of equine arteritis virus (EAV) and 44% amino acid identity to those of the 1b proteins of coronaviruses and Berne virus. Combined, the results indicate that LDV replication involves formation of a 3'-coterminal-nested set of mRNAs as observed for coronaviruses and toroviruses as well as for EAV, with which LDV shares many other properties. Overall, LDV, like EAV, possesses a genome organization resembling that of the coronaviruses and toroviruses. However, EAV and LDV differ from the latter in the size of their genomes, virion size and structure, nature of the structural proteins, and symmetry of the nucleocapsids.


title: Nursing, healthcare, and culture: views from Canada and Taiwan for the year 2050. Interview by Jacqueline Fawcett.

text: This column presents a dialogue with Canadian nurse scholar Mireille Lavoie and Taiwanese nurse scholar Yea-Ing Lotus Shyu, who share their own vision and their understanding of their country's vision of nursing, health-care, and quality of life in the year 2050.


title: Viral entry, lipid rafts and caveosomes.

text: Lipid rafts and caveolae are detergent-insoluble plasma membrane microdomains, involved in cellular endocytic processes and signalling. Several viruses, including a human pathogen, echovirus 1, and an extensively studied simian virus 40 utilize these domains for internalization into the host cells. Interaction of viruses with receptors on the cell surface triggers specific conformational changes of the virus particle and can give rise to signalling events, which determine the mechanisms of virus entry. After internalization via cell surface lipid rafts or caveolae, virus-containing vesicles can fuse with caveosomes, pre-existing cytoplasmic organelles, or dock on other intracellular organelles. These pathways may deliver viruses further to different cellular destinations, where the viral replication cycle then takes place. The information concerning the viral entry processes is important for understanding the details of the infections, for finding new targets for antiviral therapy and for elucidating the cellular internalization pathways in general.


title: Cyclophilin function in Cancer; lessons from virus replication.

text: Cyclophilins belong to a group of proteins that possess peptidyl prolyl isomerase activity and catalyse the cis-trans conversion of proline peptide bonds. Cyclophilin members play important roles in protein folding and as molecular chaperones, in addition to a well-established role as host factors required for completion of the virus life cycle. Members of the cyclophilin family are overexpressed in a range of human malignancies including hepatocellular cancer, pancreatic cancer, nonsmall cell lung cancer, gastric cancer, colorectal cancer and glioblastoma multiforme, however, their precise role in tumourigenesis remains unclear. In recent years, mounting evidence supports a role for prolyl isomerisation during mammalian cell division; a process with striking similarity to plasma membrane remodelling during virus replication. Here, we summarise our current understanding of the role of cyclophilins in cancer. We review the function of cyclophilins during mammalian cell division and during HIV-1 infection, and highlight common processes involving members of the ESCRT and Rab GTPase families.


title: [Feline immunodeficiency virus in Switzerland: clinical aspects and epidemiology in comparison with feline leukemia virus and coronaviruses].

text: Serum samples from 1421 domestic cats (561 healthy, 860 sick) were tested for FIV-, FeLV- and coronavirus infection. The results were stored in a computer data base and compared with epidemiologic data and clinical findings. All 3 infections were significantly more prevalent in sick than healthy cats: FIV was found in 0.7% of healthy and 3.4% of sick cats. For FeLV the prevalence was 3.0% and 13.0% and for coronavirus 21% and 36.2%, respectively. FIV-infected cats were mostly male (73%); no sex predilection was observed in FeLV- and coronavirus infection. In sick cats FIV-infection was significantly more prevalent in cats greater than 2 years of age; no age-dependence was found in FeLV- and coronavirus infections. The prevalence of FIV-infection increased significantly with the number of animals per household. In contrast, the frequency of FeLV infection decreased with the number of animals per household. Prevalence of coronavirus infection did not vary with group size or living conditions. The following clinical symptoms were associated with infection: FIV: general depression, diseases of the urinary tract; FeLV: general depression, fever, rough hair coat, lymphadenopathy, impaired functions of heart and circulation and muscle atrophy; coronavirus: lymphadenopathy and alterations in the abdomen. It was concluded that based on the clinical symptoms alone FIV-infection could not be diagnosed nor differentiated from the other 2 infections.


title: Central nervous system pathology caused by autoreactive CD8+ T-cell clones following virus infection.

text: Theiler's murine encephalomyelitis virus (TMEV) causes a demyelinating disease in infected mice which has similarities to multiple sclerosis. Spleen cells from TMEV-infected SJL/J mice stimulated with antigen-presenting cells infected with TMEV resulted in a population of autoreactive CD8+ cytotoxic T cells that kill uninfected syngeneic cells. We established CD8+ T cell clones that could kill both TMEV-infected and uninfected syngeneic targets, although infected target cells were killed more efficiently. The CD8+ T-cell clones produced gamma interferon when incubated with either infected or uninfected syngeneic target cells. Intracerebral injection of the clones into naïve mice induced degeneration, not only in the brain, but also in the spinal cord. This suggests that CD8+ Tc1 cells could play a pathogenic role in central nervous system inflammation.


Aetiological role of viruses in puppies with diarrhoea.


title: Anesthetic management during surgery for left ventricular aneurysm and false aneurysm occurring in stage: a case report.

text: Left ventricular aneurysm (LVA) and false aneurysm are complications of acute myocardial infarction, trauma, and cardiac surgery. Left ventricular false aneurysm (LVFA) is a particularly catastrophic complication owing to its high propensity for rupture. Surgical resection should be considered for LVFA occurring within three months after myocardial infarction or development of congestive heart failure. In this report, we describe a case of acute heart failure with LVA and LVFA occurring in stage as a complication of myocardial infarction in a 55-year-old man. The patient was also at risk of brain ischemia due to abnormal vessel status and a previous cerebrovascular accident with left-sided weakness. Successful perioperative anesthetic management was achieved by focusing on maintaining marginal upper normal blood pressure to ensure cerebral perfusion and to reduce the risk of false aneurysm rupture.


title: Fatal infection with feline panleukopenia virus in two captive wild carnivores (Panthera tigris and Panthera leo).

text: Two cases of fatal infection caused by parvovirus in a white tiger (Panthera tigris) and an African lion (Panthera leo) at the Lisbon Zoo (Portugal) are described. Gross findings at necropsy were catharral enteritis in the tiger and severe hemorrhagic enteritis in the lion. Histopathologic examination revealed, in both animals, intestinal crypt necrosis and lymphocyte depletion in the germinal centers of the mesenteric lymph nodes. Bacteriologic examination was negative for common bacterial pathogens, including Salmonella. Amplification of the parvovirus VP2 complete gene was achieved in both cases and sequencing analysis identified these isolates as feline panleukopenia virus (FPLV). The nucleotide sequences obtained from these two viruses were genetically indistinguishable. The phylogenetic analysis of FPLV strains from domestic cats obtained in the Lisbon area revealed the circulation of FPLV strains highly similar to those isolated from the tiger and lion, which strongly suggests that stray cats may have been the source of infection.


title: Coronavirus envelope glycoprotein assembly complexes.

text: Protein:protein interactions, and their subcellular localization, play important roles in coronavirus assembly. In this study, we have identified similar envelope glycoprotein complexes that are present in mouse hepatitis coronavirus A59 (MHV-A59) and bovine coronavirus (BCV) infected cells. Complexes consisting of the spike (S) and membrane (M) proteins were identified in cells infected with MHV-A59 or BCV. Kinetic analyses demonstrated that S and M quickly associated after translation, and suggested that both initially interacted in a pre-Golgi site. In addition, the hemagglutinin esterase (HE) was identified as part of a complex with M and S in BCV infected cells. Taken together, our data indicate that similar glycoprotein complexes are present in cells infected with two different coronaviruses, and thus likely represent important prerequisite complexes involved in virus assembly.


title: [The cloning, expression and structural analysis of putative unknown protein Orf 9b in SARS-CoV].

text: Orf 9b was amplified by PCR from SARS-CoV genome and cloned into the Nco I and Bam HI sites of the pET32c expression vector, and then recombinant plasmid pET32c-Orf 9b was constructed. The recombinant fusion protein Orf 9b was expressed by IPTG induction and purifed. After being cleaved by rEK, Orf 9b protein with MW 11 kD was separated and collected. It was demonstrated by ELISA that the purified Orf 9b protein could react with sera of SARS rehabilitaion patients but not with sera from healthy donors. CD and Infrared spectroscopy were used to predict the secondary structure of the purified Orf 9b protein. The distribution percentages for the the secondary structures of alpha-helix, beta-sheet, and random coil in the Orf 9b protein estimated by CD were 12.5%, 40%, and 47.5%, respectively, while the same parameters estimated by Infrared spectroscopy were 13.7%, 47.5%, and 37.9%, respectively. The results obtained by the two methods were substantially identical and showed that the structure of the Orf 9b protein consisted mostly of beta-sheet, and comprised only few alpha-helix. The acquisition of purified protein and the structural information presented herein may provide foundation for further functional study.


title: Diagnosis of Giardia infection in 14 calves.

text: Fourteen calves, 12 days to 12 weeks old, were treated for diarrhea. Fecal examination for parasitologic, bacteriologic, or viral infection revealed Giardia in all calves; rotavirus and coronavirus were found in some calves. Thirteen affected calves were treated orally with dimetridazole (50 mg/kg of body weight, daily, for 5 days), with complete resolution of the diarrhea and elimination of the Giardia. Giardiasis should be considered as an etiologic agent of diarrhea in calves.


title: Coronavirus IBV: further evidence that the surface projections are associated with two glycopolypeptides.

text: The surface projections (peplomers) of avian infectious bronchitis virus (IBV) strain M41 have been separated from the nucleocapsid (N) and matrix (M) proteins by sedimentation in a sucrose gradient after virus disruption by the non-ionic detergent Nonidet P40. The peplomers comprised two glycopolypeptides of mol. wt. 90 X 10(3) (90K; S1) and 84K (S2), shown by analysis of differentially radiolabelled virus to be present in equimolar proportions. Polypeptides of 75K and 110K, which were detected by Coomassie Brilliant Blue staining in similar amounts to S1 and S2 in some unlabelled virus preparations, were absent from peplomer preparations and are probably host cell polypeptides. The S1:S2:N:M polypeptide molar ratio for IBV-M41 was approximately 1:1:6:15.


New Zealand's elimination strategy for the COVID-19 pandemic and what is required to make it work.


title: Otitis media with effusion.

text: Otitis media with effusion is common in young children and varies widely in duration and severity. Defined as fluid in the middle ear without signs or symptoms of infection, it is generally asymptomatic. However, it has the potential to adversely affect hearing, speech development, and language acquisition.


title: Coronaviruses associated with outbreaks of transmissible enteritis of turkeys in Quebec: hemagglutination properties and cell cultivation.

text: Coronaviruses were observed by electron microscopy in the intestinal contents of turkeys in Quebec flocks where repeated outbreaks of enteritis occurred. Three isolates could be serially propagated in turkey embryos inoculated by the amniotic route with clarified intestinal contents. Purification and concentration of viral particles contained in intestinal contents of infected embryos were achieved by precipitation with polyethylene glycol and ultracentrifugation on sucrose density gradients. Three particle types were demonstrated: intact virions with a density of 1.18 to 1.20 g/ml and incomplete particles with densities of 1.14 and 1.24 g/ml. Hemagglutination of rabbit and guinea pig erythrocytes was demonstrated with the intact viral particles; the hemagglutinin was not dependent on incubation temperature. All the isolates were antigenically related, as shown by hemagglutination-inhibition. The turkey coronaviruses did not cross-react with antisera against coronaviruses of avian infectious bronchitis, porcine transmissible enteritis, bovine neonatal calf diarrhea, or mouse hepatitis. One of the Quebec isolates was shown to induce syncytia formation on its third passage in primary chicken-embryo kidney cell cultures. Electron-microscopic examination of infected cell-culture fluids revealed characteristics coronavirus particles identical to those found in intestinal contents of infected turkeys.


Intracellular development and mechanism of hemadsorption of a human coronavirus, OC43.


title: Transmissible gastroenteritis virus and porcine respiratory coronavirus: molecular characterization of the S gene using cDNA probes and nucleotide sequence analysis.

text: Two transmissible gastroenteritis virus (TGEV, Miller strain) cDNA clones were identified and their nucleotide sequences determined. The clones were non-overlapping and were located in the 5' region of the S glycoprotein gene. The TGEV clone pE21 contained 381 bp of the S glycoprotein gene and had > 98% nucleotide and amino acid sequence homology with the Purdue (P115) strain of TGEV and over 87% sequence homology with feline infectious peritonitis virus (FIPV). The TGEV clone, pD24, contained 267 bp of the S glycoprotein gene. It had > 98% nucleotide and amino acid sequence homology with P115 but only a 49% nucleotide sequence homology and a 24% amino acid sequence homology with FIPV. Using dot blot hybridization, a probe prepared from pD24 could differentiate TGEV from the antigenically related coronaviruses, FIPV, feline enteric coronavirus and canine coronavirus. This probe could also differentiate TGEV from porcine respiratory coronavirus (PRCV). Using polymerase chain reaction amplified regions of PRCV isolates and nucleotide sequencing, a 681 bp deletion in the 5' region of the S gene from PRCV isolate ISU-1 was identified. This deletion was located in the area of the S glycoprotein gene identified by the pD24 probe.


title: Zinc gluconate lozenges for treating the common cold in children: a randomized controlled trial.

text: CONTEXT The common cold is one of the most frequently occurring illnesses and is responsible for substantial morbidity and economic loss. Biochemical evidence suggests that zinc may be an effective treatment, and zinc gluconate glycine (ZGG) lozenges have been shown to reduce the duration of cold symptoms in adults. OBJECTIVE To determine the efficacy of ZGG treatment of colds in children and adolescents. DESIGN A randomized, double-masked, placebo-controlled study. SETTING Two suburban school districts in Cleveland, Ohio. PATIENTS A total of 249 students in grades 1 through 12 were enrolled within the first 24 hours of experiencing at least 2 of 9 symptoms of the common cold. INTERVENTION Zinc lozenges, 10 mg, orally dissolved, 5 times a day (in grades 1-6) or 6 times a day (in grades 7-12). MAIN OUTCOME MEASURES Time to resolution of cold symptoms based on subjective daily symptom scores for cough, headache, hoarseness, muscle ache, nasal congestion, nasal drainage, scratchy throat, sore throat, and sneezing. RESULTS Time to resolution of all cold symptoms did not differ significantly between students receiving zinc (n = 124) and those receiving placebo (n = 125) (median, 9 days; 95% confidence interval [CI], 8-9 days; median, 9 days, 95% CI, 7-10 days, respectively; P=.71). There were no significant differences in the time to resolution of any of the 9 symptoms studied. Compared with controls, more students in the zinc group reported adverse effects (88.6% vs 79.8%; P=.06); bad taste (60.2% vs 37.9%; P=.001); nausea (29.3% vs 16.1%; P=.01); mouth, tongue, or throat discomfort (36.6% vs 24.2%; P=.03); and diarrhea (10.6% vs 4.0%; P=.05). CONCLUSIONS In this community-based, randomized controlled trial, ZGG lozenges were not effective in treating cold symptoms in children and adolescents. Further studies with virologic testing are needed to clarify what role, if any, zinc may play in treating cold symptoms.


title: Radiofrequency ablation of lung cancer at Okayama University Hospital: a review of 10 years of experience.

text: The application of radiofrequency ablation for the treatment of lung cancer by our group at Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences began in June 2001, and in the present report, we review our 10-year experience with this treatment modality at Okayama University Hospital. The local efficacy of radiofrequency ablation for the treatment of lung cancer depends on tumor size and the type of electrode used, but not on tumor type. An important factor for the prevention of local failure may be the acquisition of an adequate ablative margin. The combination of embolization and radiation therapy enhances the local efficacy. Local failure may be salvaged by repeating the radiofrequency ablation, particularly in small tumors. Survival rates after radiofrequency ablation are quite promising for patients with clinical stage I non-small cell lung cancer and pulmonary metastasis from colorectal cancer, hepatocellular carcinoma, and renal cell carcinoma. The complications caused by radiofrequency ablation can be treated conservatively in the majority of cases. However, attention should be paid to rare but serious complications. This review shows that radiofrequency ablation is a promising treatment for patients with lung cancer.


title: Harnessing fetal and adult genetic reprograming for therapy of heart disease.

text: Heart is the first organ formed during organogenesis. The fetal heart undergoes several structural and functional modifications to form the four-chambered mammalian heart. The adult heart shows different adaptations during compensatory and decompensatory heart failure. However, one common adaptation in the pathological heart is fetal reprogramming, where the adult heart expresses several genes and miRNAs which are active in the fetal stage. The fetal reprogramming in the failing heart raises several questions, such as whether the switch of adult to fetal genetic programming is an adaptive response to cope with adverse remodeling of the heart, does the expression of fetal genes protect the heart during compensatory and/or decompensatory heart failure, does repressing the fetal gene in the failing heart is protective to the heart? To answer these questions, we need to understand the expression of genes and miRNAs that are reprogrammed in the failing heart. In view of this, we provided an overview of differentially expressed genes and miRNAs, and their regulation in this review. Further, we elaborated novel strategies for a plausible future therapy of cardiovascular diseases.


Responses to steroids and bronchodilators in COPD in the ISOLDE trial: the fat lady sings on.


title: Heterogeneity detector: finding heterogeneous positions in Phred/Phrap assemblies.

text: UNLABELLED A modification to Phred and program to detect heterogeneous positions, which is particularly useful in the identification of mutations and other abnormalities in Phred/Phrap genome assemblies. AVAILABILITY The package is made available at http://glscompute.gis.a-star.edu.sg/~charlie/DHetero.html


title: The Impact of Roller Pump vs. Centrifugal Pump on Homologous Blood Transfusion in Pediatric Cardiac Surgery.

text: Centrifugal pumps are considered to be less destructive to blood elements (1) when compared to roller pumps. However, their large prime volumes render them unsuitable as arterial pumps in heart lung machine (HLM) circuitry for children. In November of 2014, the circuit at Arnold Palmer Hospital, a Biomedicus BP-50 with kinetic assist venous drainage (KAVD) and 1/4″ tubing was converted to a roller pump in the arterial position with gravity drainage. Vacuum-assisted venous drainage (VAVD) was mounted on the HLM as a backup, but not used. Tubing was changed to 3/16″ in the arterial line in patients <13 kg. A retrospective study with a total of 140 patients compared patients placed on cardiopulmonary bypass (CPB) with Biomedicus centrifugal pumps and KAVD (Centrifugal Group, n = 40) to those placed on CPB with roller pumps and gravity drainage (Roller Group, n = 100). Patients requiring extra-corporeal membrane oxygenation (ECMO)/cardio-pulmonary support (CPS) or undergoing a hybrid procedure were excluded. Re-operation or circulatory arrest patients were not excluded. Prime volumes decreased by 57% from 456 ± 34 mL in the Centrifugal Group to 197 ± 34 mL in the Roller Group (p < .001). There was a corresponding increase in hematocrit (HCT) of blood primes and also on CPB. Intraoperative homologous blood transfusions also decreased 55% from 422 mL in the Centrifugal Group to 231 mL in the Roller Group (p < .001). The Society of Thoracic Surgeons--European Association for Cardio-Thoracic Surgery (STAT) categorized intubation times and hospital length of stay (LOS) for all infants showed a trend toward reduction, but was not statistically significant. Overall mortality was 5% utilizing the centrifugal configuration and 0% in the roller pump cohort. We demonstrated that the transition to roller pumps in the arterial position of the HLM considerably reduced our priming volume and formed a basis for a comprehensive blood conservation program. By maintaining higher HCTs on CPB, we were able to reduce intraoperative homologous blood transfusions.


title: Nanomaterials and nanoparticles: sources and toxicity.

text: This review is presented as a common foundation for scientists interested in nanoparticles, their origin,activity, and biological toxicity. It is written with the goal of rationalizing and informing public health concerns related to this sometimes-strange new science of "nano," while raising awareness of nanomaterials' toxicity among scientists and manufacturers handling them.We show that humans have always been exposed to tiny particles via dust storms, volcanic ash, and other natural processes, and that our bodily systems are well adapted to protect us from these potentially harmful intruders. There ticuloendothelial system, in particular, actively neutralizes and eliminates foreign matter in the body,including viruses and nonbiological particles. Particles originating from human activities have existed for millennia, e.g., smoke from combustion and lint from garments, but the recent development of industry and combustion-based engine transportation has profoundly increased an thropogenic particulate pollution. Significantly, technological advancement has also changed the character of particulate pollution, increasing the proportion of nanometer-sized particles--"nanoparticles"--and expanding the variety of chemical compositions. Recent epidemiological studies have shown a strong correlation between particulate air pollution levels, respiratory and cardiovascular diseases, various cancers, and mortality. Adverse effects of nanoparticles on human health depend on individual factors such as genetics and existing disease, as well as exposure, and nanoparticle chemistry, size, shape,agglomeration state, and electromagnetic properties. Animal and human studies show that inhaled nanoparticles are less efficiently removed than larger particles by the macrophage clearance mechanisms in the lungs, causing lung damage, and that nanoparticles can translocate through the circulatory, lymphatic, and nervous systems to many tissues and organs, including the brain. The key to understanding the toxicity of nanoparticles is that their minute size, smaller than cells and cellular organelles, allows them to penetrate these basic biological structures, disrupting their normal function.Examples of toxic effects include tissue inflammation, and altered cellular redox balance toward oxidation, causing abnormal function or cell death. The manipulation of matter at the scale of atoms,"nanotechnology," is creating many new materials with characteristics not always easily predicted from current knowledge. Within the nearly limitless diversity of these materials, some happen to be toxic to biological systems, others are relatively benign, while others confer health benefits. Some of these materials have desirable characteristics for industrial applications, as nanostructured materials often exhibit beneficial properties, from UV absorbance in sunscreen to oil-less lubrication of motors.A rational science-based approach is needed to minimize harm caused by these materials, while supporting continued study and appropriate industrial development. As current knowledge of the toxicology of "bulk" materials may not suffice in reliably predicting toxic forms of nanoparticles,ongoing and expanded study of "nanotoxicity" will be necessary. For nanotechnologies with clearly associated health risks, intelligent design of materials and devices is needed to derive the benefits of these new technologies while limiting adverse health impacts. Human exposure to toxic nanoparticles can be reduced through identifying creation-exposure pathways of toxins, a study that may someday soon unravel the mysteries of diseases such as Parkinson's and Alzheimer's. Reduction in fossil fuel combustion would have a large impact on global human exposure to nanoparticles, as would limiting deforestation and desertification.While nanotoxicity is a relatively new concept to science, this review reveals the result of life's long history of evolution in the presence of nanoparticles, and how the human body, in particular, has adapted to defend itself against nanoparticulate intruders.


title: LAPAROSCOPIC LEFT LATERAL SECTIONECTOMY AT DR GEORGE MUKHARI ACADEMIC HOSPITAL.

text: BACKGROUND Laparoscopic left lateral sectionectomy has rapidly become a standard of care for patients requiring excision of segments 2 and 3. We present our technique of the resection. METHOD Videos of patients having had left lateral sectionectomies were reviewed. Patients were placed in supine with reverse Trendelenburg position. Three ports were used: one 12 mm at the umbilicus for the camera, one 12 mm placed 5 cm above and to the right of the umbilicus and one 5 mm port to the left. A Ligasure® 5 mm (Covidien®) was used to take down the falciform ligament initially staying close to the abdominal wall and then stopping at the bifurcation of the two layers of peritoneum. The triangular ligament was incised to create a window between diaphragm and liver. Unlike open surgery the entire ligament was not mobilised at this stage. A line was scored on the left of the falciform ligament. Transection of the parenchyma was achieved with Ligasure®. Once the liver parenchyma was thinned, an endostapler - EndoGIA® Gold (Covidien®) - was be used to complete the transection. It was also possible to dissect out individual vessels and ligate with Hem-o-lok® (Teleflex®) clips in one case. The cut surface was examined for bleeding or bile leaks. A small pencil drain was placed. RESULTS Three patients had laparoscopic resections. All were female. Two cases were haemangioma with one diagnosed preoperatively and the other not having typical features on imaging. The third case was a large adenoma in a patient with hepatic adenomatosis. No bile leaks were noted. No patient required blood transfusion. Average length of stay was 1 day. CONCLUSION Laparoscopic left lateral sectionectomy is a stepping stone to more complex laparoscopic liver resections. It is feasible and safe in a tertiary institute in South Africa.


title: Mutant murine hepatitis virus-induced apoptosis in the hippocampus.

text: The mutant virus Mu-3 was isolated from the soluble receptor-resistant mutant 7 virus (srr7), which is a neuropathogenic strain of the mouse hepatitis virus JHMV, and cloned as a soluble receptor-resistant mutant from the highly neuropathogenic JHMV strain cl-2 virus (cl-2). In order to identify specific characteristics of Mu-3, the pathology of Mu-3-infected mice was compared with that of srr7- and cl-2-infected mice. The neuropathology after Mu-3 infection exhibited a mixed pattern comparable to that induced by srr7 and cl-2 infections. In addition, Mu-3 infection caused marked apoptotic lesions in the hippocampal region, particularly in the CA2 and CA3 subregions, in the brains of all infected mice. In contrast, in cl-2 infection, 10-20% of the infected mice exhibited apoptosis in the hippocampus, which was primarily observed in the CA1 subregion. Apoptosis also occurred in the pyramidal neurons and CD11b-bearing cells. The apoptotic cells, indicated by caspase 3-activation, were a mixed population of infected and a higher number of uninfected cells. These data indicated that apoptosis observed in Mu-3 infection could be induced by the indirect effects of infection in addition to direct effects of the infected cells occurring in a cell-autonomous manner.


The RNA and proteins of human coronaviruses.


title: The significance of interleukin-6 and lactate in the synovial fluid for diagnosing native septic arthritis.

text: Aim of this study was to evaluate the role of synovial interleukin-6 and synovial lactate for predicting native septic arthritis. We analyzed retrospectively synovial fluid parameters (interleukin-6, total-protein, glucose, lactate, synovial-fluid-white-blood-cell-count) of 62 patients with culture-verified native septic arthritis and compared them to 57 patients with acute aseptic arthritis. Receiver-Operating-Characteristic-curves were calculated to determine the 'Area-under-the-curves' (AUC), the best thresholds and the corresponding likelihood-ratios. The best parameter for diagnosing septic arthritis was synovial lactate (AUC = 0.864, sensitivity = 74.5%, specificity = 87.2%), followed by synovial interleukin-6 (AUC = 0.803, sensitivity = 92.5%, specificity = 64.1%) and the synovial-fluid-white-blood-cell-count (AUC = 0.782, sensitivity = 71.2%, specificity = 84.9%). Synovial lactate levels above 10 mmol/l almost proofed septic arthritis (interval-Likelihood-Ratio = 20.4), synovial interleukin-6 levels lower than 7000 pg/ml almost ruled out infection (interval-Likelihood-Ratio = 0.12). If none of these thresholds are met, physicians should estimate disease probability by the simultaneous use of the interval-Likelihood-Ratios of synovial lactate, synovial interleukin-6 and synovial-fluid-white-blood-cell-count.


title: Identification of respiratory virus in infants with congenital heart disease by comparison of different methods.

text: Respiratory virus infections are the main cause of infant hospitalization and are potentially severe in children with congenital heart disease (CHD). Rapid and sensitive diagnosis is very important to early introduction of antiviral treatment and implementation of precautions to control transmission, reducing the risk of nosocomial infections. In the present study we compare different techniques in the diagnosis of respiratory viruses in CHD infants. Thirty-nine samples of nasopharyngeal aspirate were obtained from CHD infants with symptoms of respiratory infection. The Multiplex PCR (Seeplex® RV 12 ACE Detection) driven to the detection of 12 respiratory viruses was compared with the direct immunofluorescence assay (DFA) and PCR, both targeting seven respiratory viruses. The positivity found by DFA, Multiplex and PCR was 33.3%, 51.3% and 48.7%, respectively. Kappa index comparing DFA and Multiplex, DFA and PCR and PCR and Multiplex PCR was 0.542, 0.483 and 0.539, respectively. The concordance between techniques was considered moderate. Both Multiplex PCR (p = 0.001) and PCR (p = 0.002) detected significantly more respiratory virus than DFA. As the performance of the tests may vary, the combination of two or more techniques may increase diagnostic sensitivity favoring the diagnosis of co-infections, early introduction of antiviral therapy and implementation of appropriate measures.


Translation of the MHV sM protein is mediated by the internal entry of ribosomes on mRNA 5.


title: The role of superantigens of group A Streptococcus and Staphylococcus aureus in Kawasaki disease.

text: PURPOSE OF REVIEW Since the first suggestion of a superantigen hypothesis for Kawasaki disease over a decade ago, debate on the aetiology remains inconclusive. This article reviews recent publications that address the role of superantigens of group A Streptococcus and Staphylococcus aureus in the pathogenesis of Kawasaki disease. RECENT FINDINGS Over the past few years, new superantigens produced by group A Streptococcus and S. aureus have been increasingly identified, bringing the total known number to more than 30. Several studies on T-cell Vbeta repertoires and seroloepidemiology have demonstrated evidence for the involvement of single or multiple superantigens produced by the two pathogens. The associated superantigens differed in those studies, including streptococcal pyrogenic toxins A and C, staphylococcal enterotoxins A-C, and toxic shock syndrome toxin 1. These disparate findings suggest that the inflammation of Kawasaki disease does not result from a single agent but rather a final common inflammatory pathway in genetically susceptible individuals after numerous infectious agents. SUMMARY Certain staphylococcal and streptococcal superantigens are suggested to be responsible for the development of Kawasaki disease. A better understanding of the precise role of the causative agents will lead to accurate diagnosis, more targeted therapy and an improvement of coronary outcomes.


Cloning of a transmissible gastroenteritis coronavirus full-length cDNA.


title: The economic burden of non-influenza-related viral respiratory tract infection in the United States.

text: BACKGROUND Viral respiratory tract infection (VRTI) is the most common illness in humans. Despite the high incidence, the economic impact of non-influenza-related VRTI has not been rigorously explored. Our objectives were to obtain an updated incidence of non-influenza-related VRTI in the United States and to quantify the health care resource use (direct costs) and productivity losses (indirect costs) associated with these infections. METHODS A nationwide telephone survey of US households (N = 4051) was conducted between November 3, 2000, and February 12, 2001 to obtain a representative estimate of the self-reported incidence of non-influenza-related VRTI and related treatment patterns. Direct treatment costs measured included outpatient clinician encounters, use of over-the-counter and prescription drugs, and associated infectious complications of non-influenza-related VRTI. Absenteeism estimates for infected individuals and parents of infected children were extrapolated from National Health Interview Survey data. RESULTS Of survey respondents, 72% reported a non-influenza-related VRTI within the past year. Respondents who experienced a self-reported non-influenza-related VRTI averaged 2.5 episodes annually. When these rates are extrapolated to the entire US population, approximately 500 million non-influenza-related VRTI episodes occur per year. Similarly, if the treatment patterns reported by the respondents are extended to the population, the total economic impact of non-influenza-related VRTI approaches $40 billion annually (direct costs, $17 billion per year; and indirect costs, $22.5 billion per year). CONCLUSIONS Largely because of the high attack rate, non-influenza-related VRTI imposes a greater economic burden than many other clinical conditions. The pending availability of effective antiviral therapies warrants increased attention be paid to this common and expensive illness.


title: Intracranial epidural mucocele in a cat.

text: An 18-month-old, spayed female, domestic shorthaired cat was presented with clinical signs of depression and reluctance to walk, which progressed to nonambulatory tetraparesis. Increased opacification of both frontal sinuses and a cyst-like abnormality causing compression and displacement of the right frontal lobe were seen on computed tomography. Bilateral frontal sinus trephination and right transfrontal craniotomy revealed clear, viscous fluid in the right frontal sinus and rostral fossa, compatible with an intracranial mucocele. At a 6-month follow-up examination, no signs of recurrence were appreciated.


title: Modes of action of disinfectants.

text: The exact mechanism of action of a disinfectant is not easy to elucidate. The notion of 'target' in the bacterial cell, frequently evoked for the antibiotics, is not clear for disinfectants (except for some, e.g. chlorhexidine). In understanding the mode of action of a disinfectant, it can be difficult to distinguish the primary stage (characteristic of the mode of action) and the secondary stage (consequence of the action). The author describes the actions of disinfectants on the external membrane, cytoplasmic membrane and energy metabolism of cells; these actions include rupture of the membrane, loss of permeability and coagulation of the cytoplasm.


title: The fitness of defective interfering murine coronavirus DI-a and its derivatives is decreased by nonsense and frameshift mutations.

text: The genome of the defective interfering (DI) mouse hepatitis virus DI-a carries a large open reading frame (ORF) consisting of ORF1a, ORF1b, and nucleocapsid sequences. To test whether this fusion ORF is important for DI virus replication, we constructed derivatives of the DI-a genome in which the reading frame was truncated by a nonsense codon or a frameshift mutation. In vitro-transcribed DI RNAs were transfected into mouse hepatitis virus-infected cells followed by undiluted passage of the resulting virus-DI virus stocks. The following observations were made. (i) Truncation of the fusion ORF was not lethal but led to reduced accumulation of DI RNA. (ii) When pairs of nearly identical in-frame and out-of-frame DI RNAs were directly compared by cotransfection, DI viruses containing in-frame genomic RNAs prevailed within three successive passage even when the out-of-frame RNAs were transfected in 10-fold molar excess. (iii) When DI viruses containing out-of-frame genomic RNAs were passaged, mutants emerged and were selected for that had restored the reading frame. We conclude that translation of the fusion ORF is indeed required for efficient propagation of DI-a and its derivatives.


title: Native nucleic acid electrophoresis as an efficient alternative for genotyping method of influenza virus.

text: Influenza viruses are the worldwide major causative agents of human and animal acute respiratory infections. Some of the influenza subtypes have caused epidemics and pandemics among humans. The varieties of methods are available for the rapid isolation and identification of influenza viruses in clinical and environmental samples. Since nucleic acids amplification techniques such as RT-PCR have been adapted, fast and sensitive influenza type and subtype determination is possible. However, in some ambiguous cases other, more detailed assay might be desired. The genetic material of influenza virus is highly unstable and constantly mutates. It is known that single nucleotide polymorphisms (SNPs) results in resistance to commercially available anti-viral drugs. The genetic drift of the virus could also result in weakening of immune response to infection. Finally, in a substantial number of patients co-infection with various virus strains or types has been confirmed. Although the detection of co-infection or presence of minor genetic variants within flu-infected patients is not a routine procedure, a rapid and wide spectrum diagnostics of influenza virus infections could reveal an accurate picture of the disease and more importantly, is crucial for choosing the appropriate therapeutics and virus monitoring. Herein we present the evidences that native gel electrophoresis and MSSCP--a method based on multitemperature single strand conformation polymorphism could furnish a useful technique for minor variants, which escape discovery by conventional diagnostic assays.


title: Effects of osteocytes on vibration-induced reflex muscle activity in postmenopausal women.

text: BACKGROUND/AIM To assess whether osteocytes have an effect on reflex myoelectrical activity during whole-body vibration (WBV) in postmenopausal women. MATERIALS AND METHODS Participants were classified into 2 groups: the low bone mineral density (BMD) group (n = 37) and normal BMD group (n = 43). Hip BMD was measured using dual-energy X-ray absorptiometry. Surface electromyography data recorded from the adductor longus muscle were processed to obtain vibration-induced reflex myoelectrical activity. Changes in plasma sclerostin (SOST) levels with WBV were expressed as a standardized vibration-induced SOST index. RESULTS The standardized vibration-induced SOST index was 1.03 ± 0.24 in the low BMD group and 0.99 ± 0.33 in the normal BMD group. For plasma SOST levels, no group-by-time interaction was found. The resting myoelectrical activities of adductor muscles increased significantly during WBV in both groups. However, there was no significant difference in the main effects of WBV on resting myoelectrical activity between the groups. The standardized vibration-induced plasma SOST index was found to be a significant independent predictor of the standardized vibration-induced reflex myoelectrical activity of the adductor muscle in both groups. CONCLUSION This study suggests that osteocytes serve as mechanoreceptors of reflex electromyography during WBV.


WHO lacks teeth on international health issues, says professor.


title: PFDB: a generic protein family database integrating the CATH domain structure database with sequence based protein family resources.

text: MOTIVATION The PFDB (Protein Family Database) is a new database designed to integrate protein family-related data with relevant functional and genomic data. It currently manages biological data for three projects-the CATH protein domain database (Orengo et al., 1997; Pearl et al., 2001), the VIDA virus domains database (Albà et al., 2001) and the Gene3D database (Buchan et al., 2001). The PFDB has been designed to accommodate protein families identified by a variety of sequence based or structure based protocols and provides a generic resource for biological research by enabling mapping between different protein families and diverse biochemical and genetic data, including complete genomes. RESULTS A characteristic feature of the PFDB is that it has a number of meta-level entities (for example aggregation, collection and inclusion) represented as base tables in the final design. The explicit representation of relationships at the meta-level has a number of advantages, including flexibility-both in terms of the range of queries that can be formulated and the ability to integrate new biological entities within the existing design. A potential drawback with this approach-poor performance caused by the number of joins across meta-level tables-is avoided by implementing the PFDB with materialized views using the mature relational database technology of Oracle 8i. The resultant database is both fast and flexible. This paper presents the principles on which the database has been designed and implemented, and describes the current status of the database and query facilities supported.


title: Modular structure of the receptor binding proteins of Lactococcus lactis phages. The RBP structure of the temperate phage TP901-1.

text: Lactococcus lactis is a gram-positive bacterium widely used by the dairy industry. Several industrial L. lactis strains are sensitive to various distinct bacteriophages. Most of them belong to the Siphoviridae family and comprise several species, among which the 936 and P335 are prominent. Members of these two phage species recognize their hosts through the interaction of their receptor-binding protein (RBP) with external cell wall saccharidices of the host, the "receptors." We report here the 1.65 A resolution crystal structure of the RBP from phage TP901-1, a member of the P335 species. This RBP of 163 amino acids is a homotrimer comprising three domains: a helical N terminus, an interlaced beta-prism, and a beta-barrel, the head domain (residues 64-163), which binds a glycerol molecule. Fluorescence quenching experiments indicated that the RBP exhibits high affinity for glycerol, muramyl-dipeptide, and other saccharides in solution. The structural comparison of this RBP with that of lactococcal phage p2 RBP, a member of the 936 species (Spinelli, S., Desmyter, A., Verrips, C. T., de Haard, J. W., Moineau, S., and Cambillau, C. (2006) Nat. Struct. Mol. Biol. 13, 85-89) suggests a large extent of modularity in RBPs of lactococcal phages.


title: The membrane M protein carboxy terminus binds to transmissible gastroenteritis coronavirus core and contributes to core stability.

text: The architecture of transmissible gastroenteritis coronavirus includes three different structural levels, the envelope, an internal core, and the nucleocapsid that is released when the core is disrupted. Starting from purified virions, core structures have been reproducibly isolated as independent entities. The cores were stabilized at basic pH and by the presence of divalent cations, with Mg(2+) ions more effectively contributing to core stability. Core structures showed high resistance to different concentrations of detergents, reducing agents, and urea and low concentrations of monovalent ions (<200 mM). Cores were composed of the nucleoprotein, RNA, and the C domain of the membrane (M) protein. At high salt concentrations (200 to 300 mM), the M protein was no longer associated with the nucleocapsid, which resulted in destruction of the core structure. A specific ionic interaction between the M protein carboxy terminus and the nucleocapsid was demonstrated using three complementary approaches: (i) a binding assay performed between a collection of M protein amino acid substitution or deletion mutants and purified nucleocapsids that led to the identification of a 16-amino-acid (aa) domain (aa 237 to 252) as being responsible for binding the M protein to the nucleocapsid; (ii) the specific inhibition of this binding by monoclonal antibodies (MAbs) binding to a carboxy-terminal M protein domain close to the indicated peptide but not by MAbs specific for the M protein amino terminus; and (iii) a 26-residue peptide, including the predicted sequence (aa 237 to 252), which specifically inhibited the binding. Direct binding of the M protein to the nucleoprotein was predicted, since degradation of the exposed RNA by RNase treatment did not affect the binding. It is proposed that the M protein is embedded within the virus membrane and that the C region, exposed to the interior face of the virion in a population of these molecules, interacts with the nucleocapsid to which it is anchored, forming the core. Only the C region of the M protein is part of the core.


title: [Development and evaluation of the kit for detection of SARS-associated Coronavirus RNA].

text: AIM To develop a diagnostic kit for detection of SARS (severe acute respiratory syndrome)-related coronavirus RNA based on reverse transcription and polymerase chain reaction and to estimate its specificity and sensitivity. MATERIAL AND METHODS 68 virus and bacterial cultures, 240 clinical samples from people without SARS symptoms and also 22 RNA samples from patients with SARS symptoms received during the epidemic in Beijing were used. RESULTS The specificity of the kit was determined using animal coronaviruses and other bacterial and viral strains, causing acute respiratory and intestinal infections, and was shown to be 100%. The sensitivity of the kit in different clinical samples was 2.2 x 10(3) genome equivalents of recombinant SARS RNA in 1 ml of the specimen. The kit was evaluated in the Institute of Microbiology and Epidemiology of Beijing (China) using SARS-cov viral suspension and clinical samples from patients with suspected SARS. It was shown that kit was able to detect 10 TCID/50 ml of SARS-Cov virus. Testing of clinical samples from patients with suspected SARS showed that diagnostic sensitivity of the kit was 95%. Detection of the SARS-Cov RNA was more effective in feces compared to sputum 990 and 40%, respectively). CONCLUSION The kit "AmpliSens SARS" for qualitative detection of SARS-related coronavirus RNA by reverse transcription and polymerase chain reaction (PCR) in nasopharyngeal wash/aspirates, naso/oropharyngeal swabs, plasma, and extract from feces has been developed in the Central Research Institute for Epidemiology of the RF Ministry of Health. The kit contains reagents for RNA isolation and purification, cDNA synthesis by reverse transcription of RNA, for PCR and for electrophoretic analysis of amplified products. The kit also contains recombinant positive and internal control samples allowing to control efficiency of analysis and showed good analytical and diagnostic characteristics.


Severe acute respiratory syndrome (SARS): over 100 days into the outbreak.


title: Antigenic differentiation between transmissible gastroenteritis virus of swine and a related porcine respiratory coronavirus.

text: The antigenic relationship between a recently isolated porcine respiratory coronavirus (TLM 83) and transmissible gastroenteritis (TGE) virus of swine was studied by neutralization, immunoblotting and radioimmunoassay (RIA) using TGE virus-specific monoclonal antibodies (MAbs) and polyclonal antibodies specific for both viruses. A complete two-way neutralization activity between the two viruses was found. Immunoblotting revealed cross-reactions between TLM 83 and TGE virus antigens at the level of the envelope protein (E1), the nucleoprotein (N) and the peplomer protein (E2). By virus neutralization assays and RIA with TGE virus-specific MAbs, the presence of similar epitopes in the E1 and N proteins and in the neutralization-mediating antigenic site of the E2 protein were demonstrated. E2 protein-specific MAbs, without neutralizing activity and reacting with antigenic sites B, C and D (previously defined), failed to recognize TLM 83. These results indicated a close antigenic relationship and structural similarity between TLM 83 and TGE viruses and also suggested potential ways of differentiating between the two viruses.


title: Occupational exposure to zoonotic agents among agricultural workers in Lombardy Region, northern Italy.

text: OBJECTIVES This study was conducted in Northern Italy with the aim of defining the risk of agricultural workers' contact with biological agents through the determination of serum antibodies against selected zoonotic agents. Immunity against tetanus was also investigated. METHODS Two groups of agricultural workers consisting of 153 animal breeders (exposed) and 46 non- breeders (controls) were included in the study. In a first group of 103 workers (89 exposed and 14 controls) the serum concentrations of antibodies against Hepatitis E Virus (HEV) were measured, whereas in the second group of 96 workers (64 exposed and 32 controls) the serum concentrations of antibodies against Leptospira spp., Coxiella burnetii, Borrelia burgdorferi, Brucella spp. and Salmonella spp. were addressed. Imunization against tretanus was also studied in this group. RESULTS Animal breeders showed higher rates of IgG antibodies against Coxiella burnetii (50% vs. 31.2%), and Leptospira spp. (59.4% vs. 43.7%). Results of logistic regression analysis revealed that breeder workers showed a tendency to have higher prevalence of positivity for antibodies to Leptospira spp.and Coxiella burnetii than non-breeders (ORs ~ 3). Only one exposed subject showed antibodies against hepatitis E (none in controls), but when tested with another commercially available kit the percentage of anti HEV IgG positive subjects increased to 22.3% in the exposed, while none of the controls showed positive. None of the subjects showed antibodies against Salmonella spp. and Brucella spp. Italians and other European workers have better protection against tetanus (91%) compared to non-EU workers (81%). CONCLUSIONS The higher frequency of the presence of serum antibodies to zoonotic agents (e.g. Leptospira spp. and Coxiella burnetii) in animal breeders suggests that they are more exposed to biological agents than workers not involved in animal breeding activities. The risk of contact with HEV deserves further studies because the adoption of different assays can result in significantly different results. The promotion of immunization of agricultural workers might be a priority, in particular for migrants.


title: Multiple sclerosis and Epstein-Barr virus: a growing association.

text: The etiology of multiple sclerosis is an elusive field due to the lack of the usual cause-effect relationship model to explain a disease. In fact, with all the current evidence we start thinking in multiple sclerosis as a multi-factorial disease, where an infective agent on a genetically permissive host can lead to inflammation, demyelination and ultimately to neurological damage with dire consequences for the patients. This review will focus on the role of viruses in multiple sclerosis etiology and give some insight regarding this complicated issue.


title: Longitudinal study on respiratory viral co-infections in the presence or absence of clinical manifestation in infants aged 0-2 years.

text: We conducted a long-term follow-up study between December 2005 and February 2007 on 4 immunocompetent infants, who repeatedly presented with respiratory symptoms, using PCR-based techniques targeting 14 viruses related to acute respiratory tract infection. Of 38 specimens, 30 were collected from symptomatic infants and 8 were collected when respiratory symptoms were absent. Overall, one or more respiratory viruses were detected in 94.7% (36/38) of the specimens. Of the 36 PCR-positive specimens, 77.8% (28/36) were positive for more than one virus. Most of these co-infections were double infections (55.6% or 20/36). Of note, co-infections with 4 and 3 viruses were observed in 3 (8.3% or 3/36) and 5 (13.9% or 5/36) specimens, respectively. Of the 8 specimens collected from the 4 infants when apparent respiratory symptoms were absent, 7 (87.5%) were positive for respiratory viruses. Respiratory viral co-infections were also frequent and found in 5 of the specimens (62.5%). However, apparent correlation between disease severity and co-infection was undetectable due to the limit of the number of cases studied. Taken together, this longitudinal study revealed that respiratory viral co-infections were not infrequent in infants aged 0-2 years, regardless of the presence of respiratory symptoms (62.5-77.8%).


title: Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.

text: The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKK(250) On SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.


Sequence analysis of the 3' end (8740 nucleotides) of BECV genome; comparison with homologous MHV nucleotide sequence.


Caring for the Psychological Well-Being of Healthcare Professionals in the Covid-19 Pandemic Crisis.


title: Endosomal proteolysis by cathepsins is necessary for murine coronavirus mouse hepatitis virus type 2 spike-mediated entry.

text: Most strains of murine coronavirus mouse hepatitis virus (MHV) express a cleavable spike glycoprotein that mediates viral entry and pH-independent cell-cell fusion. The MHV type 2 (MHV-2) strain of murine coronavirus differs from other strains in that it expresses an uncleaved spike and cannot induce cell-cell fusion at neutral pH values. We show here that while infection of the prototype MHV-A59 strain is not sensitive to pretreatment with lysosomotropic agents, MHV-2 replication is significantly inhibited by these agents. By use of an A59/MHV-2 chimeric virus, the susceptibility to lysosomotropic agents is mapped to the MHV-2 spike, suggesting a requirement of acidification of endosomes for MHV-2 spike-mediated entry. However, acidification is likely not a direct trigger for MHV-2 spike-mediated membrane fusion, as low-pH treatment is unable to overcome ammonium chloride inhibition, and it also cannot induce cell-cell fusion between MHV-2-infected cells. In contrast, trypsin treatment can both overcome ammonium chloride inhibition and promote cell-cell fusion. Inhibitors of the endosomal cysteine proteases cathepsin B and cathepsin L greatly reduce MHV-2 spike-mediated entry, while they have little effect on A59 entry, suggesting that there is a proteolytic step in MHV-2 entry. Finally, a recombinant virus expressing a cleaved MHV-2 spike has the ability to induce cell-cell fusion at neutral pH values and does not require low pH and endosomal cathepsins during infection. These studies demonstrate that endosomal proteolysis by cathepsins is necessary for MHV-2 spike-mediated entry; this is similar to the entry pathway recently described for severe acute respiratory syndrome coronavirus and indicates that coronaviruses may use multiple pathways for entry.


title: Intrinsic resistance of feline peritoneal macrophages to coronavirus infection correlates with in vivo virulence.

text: Cats infected with virulent feline coronavirus strains develop feline infectious peritonitis, an invariably fatal, immunologically mediated disease; avirulent strains cause either clinically inapparent infection or mild enteritis. Four virulent coronavirus isolates and five avirulent isolates were assessed by immunofluorescence and virus titration for their ability to infect and replicate in feline peritoneal macrophages in vitro. The avirulent coronaviruses infected fewer macrophages, produced lower virus titers, were less able to sustain viral replication, and spread less efficiently to other susceptible macrophages than the virulent coronaviruses. Thus, the intrinsic resistance of feline macrophages may play a pivotal role in the outcome of coronavirus infection in vivo.


title: VaZyMolO: a tool to define and classify modularity in viral proteins.

text: Viral structural genomic projects aim at unveiling the function of unknown viral proteins by employing high-throughput approaches to determine their 3D structure and to identify their function through fold-homology studies. The 'viral enzyme module localization' (VaZyMolO) tool has been developed, which aims at defining viral protein modules that might be expressed in a soluble and functionally active form, thereby identifying candidates for crystallization studies. VaZyMolO includes 114 complete viral genome sequences of both negative- and positive-sense, single-stranded RNA viruses available from NCBI. In VaZyMolO, a module is defined as a structural and/or functional unit. Modules were first identified by homology search and then validated by the convergence of results from sequence composition analysis, motif search, transmembrane region search and domain definitions, as found in the literature. The public interface of VaZyMolO, which is accessible from http://www.vazymolo.org, allows comparison of a query sequence to all VaZyMolO modules of known function.


title: SARS revisited: managing "outbreaks" with "communications".

text: "Risk communications" has acquired some importance in the wake of our experience of SARS. Handled well, it helps to build mutual respect between a government or an organisation and the target groups with which it is communicating. It helps nurture public trust and confidence in getting over the crisis. The World Health Organization (WHO) has also come to recognise its importance after SARS and organised the first Expert Consultation on Outbreak Communications conference in Singapore in September 2004. This article assesses the context and the key features which worked to Singapore's advantage. Looking at the data now widely available on the Internet of the experience of SARS-infected countries like China, Taiwan, Canada, the article identifies the key areas of strategic communications in which Singapore fared particularly well. Another issue discussed is whether Singapore's experience has universal applicability or whether it is limited because of Singapore's unique cultural, historical and geographical circumstances. Finally, the article also looks at some of the post-SARS enhancements that have been put in place following the lessons learnt from SARS and the need to confront new infectious outbreaks like avian flu.


New data in a 200-year investigation.


title: A serologic survey of Oklahoma cats for antibodies to feline immunodeficiency virus, coronavirus, and Toxoplasma gondii and for antigen to feline leukemia virus.

text: A serologic survey was done on 618 cat sera submitted to the Oklahoma Animal Disease Diagnostic Laboratory between July 1, 1987 and June 30, 1988. The samples were collected from clinically normal and sick cats. The sera were tested for the presence of antibodies to feline immunodeficiency virus by a commercial immunoassay, to a coronavirus by an indirect fluorescent antibody test, and to Toxoplasma gondii by a commercial latex agglutination test and for the presence of feline leukemia virus antigen with one of 3 different commercial assay kits. Ten percent of the sera had antibodies to feline immunodeficiency virus, 35% had antibodies to a coronavirus, and 22% had antibodies to Toxoplasma gondii. Feline leukemia virus antigen was detected in 15% of the sera. Thirty-two percent of the sera had evidence of exposure to 2 or more of the agents.


Characterization of temperature-sensitive (ts) mutants of coronavirus infectious bronchitis virus (IBV).


Informed consent issues hobble cancer genome scheme.


title: Application of Evidence Investigator for the simultaneous measurement of soluble adhesion molecules: L-, P-, E- selectins, VCAM-1, ICAM-1 in a biochip platform.

text: The semiautomated Evidence Investigator has been applied to the simultaneous specific measurement of soluble adhesion molecules: L-, P-, E- selectins, VCAM-1, ICAM-1 using a reduced volume of sample. The biochip is the solid support and vessel where the sandwich immunoassay takes place. Signal detection, imaging, data processing, and storage are fully automated. Calibration curves are generated simultaneously for each analyte, with automatic validation against supplied calibration data. These curves are used for the calculation of the concentrations in multi-analyte controls and human serum samples. Data from the evaluation parameters assessed indicate suitability of the Evidence Investigator system for the application.


title: [Differential diagnosis of the viral etiology of suspected mumps in a population with high vaccine coverage].

text: AIM In this study, buccal swabs from patients with the clinical picture of parotitis epidemica in whom mumps virus (MV) infection was not confirmed by direct detection or serologically were tested. The aim was to detect by molecular methods nucleic acids (NAs) of other respiratory viruses possibly involved in salivary gland swelling. At the same time, paired sera, if available, were tested. MATERIAL AND METHODS The study group consisted of 72 buccal swabs from patients of the Clinic of infectious, tropical, and parasitic diseases, Na Bulovce Hospital. Paired sera were available from ten patients. Samples were collected in 2013 to 2015. Buccal swabs were tested by PCR for the presence of NAs of adenoviruses (AdV), bocaviruses (hBoV), parainfluenza viruses of types 1-4 (HPIV), human metapneumovirus (hMPV), coronaviruses (HCoV: NL63, OC43, HKU1, and 229E), respiratory syncytial virus (RSV), influenza A virus, influenza B virus, and Epstein-Barr virus (EBV). Paired sera were screened by the complement fixation test (AdV and influenza A and B viruses), hemagglutination inhibition test (HPIV types 2 and 3), ELISA (AdV, EBV), and immunofluorescence (EBV). RESULTS NAs from viruses other than the mumps virus were detected in 27 of 72 patients with clinical symptoms of parotitis epidemica, and serological tests revealed etiological links with parainfluenza viruses in three more cases. Overall, 30 (41.7%) of 72 patients with suspected mumps tested positive for one or more viruses from the study panel. The most commonly detected viruses were AdV 11/72 (15.3%), EBV 9/72 (12.5%), and HPIV 3/72 (4.2%), but influenza A virus (H3N2) 1/72 (1.4%) was also found. Some patients tested positive for more than one virus: 2/72 (3%) for AdV plus hBoV and 1/72 (1.4%) for HPIV plus HCoV. In addition, examination of paired sera revealed HPIV positivity in three more patients. PCR and serology detected etiological link with HPIV in six (8.3%) of 72 patients tested. CONCLUSION In our study group, nearly 42% of patients with the clinical picture of parotitis epidemica in whom mumps virus (MV) infection was not confirmed by direct detection or serologically tested positive for viruses other than the mumps virus. Thorough laboratory diagnosis of suspected mumps in vaccinated persons is important not only for the treatment of patients and adoption of isolation and other measures, but also for a better understanding of the epidemiology of the disease and outcomes of the immunisation programmes.


Process and outcome of care for patients with ovarian cancer.


title: The 3a protein of severe acute respiratory syndrome-associated coronavirus induces apoptosis in Vero E6 cells.

text: An outbreak of severe acute respiratory syndrome (SARS) occurred in China and the first case emerged in mid-November 2002. The aetiological agent of this disease was found to be a previously unknown coronavirus, SARS-associated coronavirus (SARS-CoV). The detailed pathology of SARS-CoV infection and the host response to the viral infection are still not known. The 3a gene encodes a non-structural viral protein, which is predicted to be a transmembrane protein. In this study, it was shown that the 3a protein was expressed in the lungs and intestinal tissues of SARS patients and that the protein localized to the endoplasmic reticulum in 3a-transfected monkey kidney Vero E6 cells. In vitro experiments of chromatin condensation and DNA fragmentation suggested that the 3a protein may trigger apoptosis. These data showed that overexpression of a single SARS-CoV protein can induce apoptosis in vitro.


title: [Autoimmune diseases in the context of pandemic COVID-19].

text: Patient suffering from autoimmune diseases (AID) typically have an increased risk of infection, which is attributed to the disease itself, but also to immunosuppressive drugs (IS) and comorbidities. During the current COVID-19 outbreak, the way to manage these diseases remains elusive. Limited data is currently available on AID and IS in the context of this new coronavirus infection. To date, there is no evidence to support an increase in complications of COVID-19 in these patients. In addition, certain drugs that are commonly used to treat AID could be part of the therapeutic arsenal used in COVID-19. The purpose of this article is to review the unique aspects of patients with AID during the COVID-19 outbreak.


[Sårbare patienter i almen praksis under COVID-19].


title: [Covid-19 diagnosis : clinical recommendations and performance of nasopharyngeal swab-PCR].

text: The Covid-19 pandemic imposes new diagnostic strategies in order to optimize the medical care of our patients. The current biblio-graphy, although of low quality, shows a sensitivity of 56 to 83 % for the Covid-19 PCR. Even though one negative test can exclude a Covid-19 in the majority of cases, the NPV (Negative Predictive Value) decreases with increasing prevalence (pre-test probability). This finding suggests the need for strict auto-isolation of patients until the resolution of their symptoms. For patients that present with typical symptoms, who have a presumed Covid-19 prevalence of -40-50 %, a negative test should be interpreted with caution and a repeat test may be needed.


title: International travels and fever screening during epidemics: a literature review on the effectiveness and potential use of non-contact infrared thermometers.

text: Several countries plan to introduce non-contact infrared thermometers (NCIT) at international airports in order to detect febrile passengers, thus to delay the introduction of a novel influenza strain. We reviewed the existing studies on fever screening by NCIT to estimate their efficacy under the hypothesis of pandemic influenza. Three Severe Acute Respiratory Syndrome (SARS) or dengue fever interventions in airports were excluded because of insufficient information. Six fever screening studies in other gathering areas, mainly hospitals, were included (N= 176 to 72,327 persons; fever prevalence= 1.2% to 16.9%). Sensitivity varied from 4.0% to 89.6%, specificity from 75.4% to 99.6%, positive predictive value (PPV) from 0.9% to 76.0% and negative predictive value (NPV) from 86.1% to 99.7%. When we fixed fever prevalence at 1% in all studies to allow comparisons, the derived PPV varied from 3.5% to 65.4% and NPV was >or=99%. The low PPV suggests limited efficacy of NCIT to detect symptomatic passengers at the early stages of a pandemic influenza, when fever prevalence among passengers would be =or<1%. External factors can also impair the screening strategy: passengers can hide their symptoms or cross borders before symptoms occur. These limits should be considered when setting up border control measures to delay the pandemic progression.


title: Videolaryngoscopy versus direct laryngoscopy for adult patients requiring tracheal intubation.

text: BACKGROUND Successful tracheal intubation during general anaesthesia traditionally requires a line of sight to the larynx attained by positioning the head and neck and using a laryngoscope to retract the tongue and soft tissues of the floor of the mouth. Difficulties with intubation commonly arise, and alternative laryngoscopes that use digital and/or fibreoptic technology have been designed to improve visibility when airway difficulty is predicted or encountered. Among these devices, a rigid videolaryngoscope (VLS) uses a blade to retract the soft tissues and transmits a lighted video image to a screen. OBJECTIVES Our primary objective was to assess whether use of videolaryngoscopy for tracheal intubation in adults requiring general anaesthesia reduces risks of complications and failure compared with direct laryngoscopy. Our secondary aim was to assess the benefits and risks of these devices in selected population groups, such as adults with obesity and those with a known or predicted difficult airway. SEARCH METHODS We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and Embase on 10 February 2015. Our search terms were relevant to the review question and were not limited by outcomes. We carried out clinical trials register searches and forward and backward citation tracking. We reran the search on 12 January 2016; we added potential new studies of interest from the 2016 search to a list of 'Studies awaiting classification', and we will incorporate these studies into the formal review during the review update. SELECTION CRITERIA We considered all randomized controlled trials and quasi-randomized studies with adult patients undergoing laryngoscopy performed with a VLS or a Macintosh laryngoscope in a clinical, emergency or out-of-hospital setting. We included parallel and cross-over study designs. DATA COLLECTION AND ANALYSIS Two review authors independently assessed trial quality and extracted data, consulting a third review author to resolve disagreements. We used standard Cochrane methodological procedures, including assessment of risk of bias. MAIN RESULTS We included 64 studies identified during the 2015 search that enrolled 7044 adult participants and compared a VLS of one or more designs with a Macintosh laryngoscope. We identified 38 studies awaiting classification and seven ongoing studies. Of the 64 included studies, 61 included elective surgical patients, and three were conducted in an emergency setting. Among 48 studies that included participants without a predicted difficult airway, 15 used techniques to simulate a difficult airway. Seven recruited participants with a known or predicted difficult airway, and the remaining studies did not specify or included both predicted and not predicted difficult airways. Only two studies specifically recruited obese participants. It was not possible to blind the intubator to the device, and we noted a high level of inevitable heterogeneity, given the large number of studies.Statistically significantly fewer failed intubations were reported when a VLS was used (Mantel-Haenszel (M-H) odds ratio (OR), random-effects 0.35, 95% confidence Interval (CI) 0.19 to 0.65; 38 studies; 4127 participants), and fewer failed intubations occurred when a VLS was used in participants with an anticipated difficult airway (M-H OR, random-effects 0.28, 95% CI 0.15 to 0.55; six studies; 830 participants). We graded the quality of this evidence as moderate on the basis of the GRADE system. Failed intubations were fewer when a VLS was used in participants with a simulated difficult airway (M-H OR, random-effects 0.18, 95% CI 0.04 to 0.77; nine studies; 810 participants), but groups with no predicted difficult airway provided no significant results (M-H OR, random-effects 0.61, 95% CI 0.22 to 1.67; 19 studies; 1743 participants).Eight studies reported on hypoxia, and only three of these described any events; results showed no differences between devices for this outcome (M-H OR, random-effects 0.39, 95% CI 0.10 to 1.44; 1319 participants). Similarly, few studies reported on mortality, noting no differences between devices (M-H OR, fixed-effect 1.09, 95% CI 0.65 to 1.82; two studies; 663 participants), and only one study reporting on the occurrence of respiratory complications (78 participants); we graded these three outcomes as very low quality owing to lack of data. We found no statistically significant differences between devices in the proportion of successful first attempts (M-H OR, random-effects 1.27, 95% CI 0.77 to 2.09; 36 studies; 4731 participants) nor in those needing more than one attempt. We graded the quality of this evidence as moderate. Studies reported no statistically significant differences in the incidence of sore throat in the postanaesthesia care unit (PACU) (M-H OR, random-effects 1.00 (95% CI 0.73 to 1.38); 10 studies; 1548 participants) nor at 24 hours postoperatively (M-H OR random-effects 0.54, 95% CI 0.27 to 1.07; eight studies; 844 participants); we graded the quality of this evidence as moderate. Data combined to include studies of cross-over design revealed statistically significantly fewer laryngeal or airway traumas (M-H OR, random-effects 0.68, 95% CI 0.48 to 0.96; 29 studies; 3110 participants) and fewer incidences of postoperative hoarseness (M-H OR, fixed-effect 0.57, 95% CI 0.36 to 0.88; six studies; 527 participants) when a VLS was used. A greater number of laryngoscopies performed with a VLS achieved a view of most of the glottis (M-H OR, random-effects 6.77, 95% CI 4.17 to 10.98; 22 studies; 2240 participants), fewer laryngoscopies performed with a VLS achieved no view of the glottis (M-H OR, random-effects 0.18, 95% CI 0.13 to 0.27; 22 studies; 2240 participants) and the VLS was easier to use (M-H OR, random-effects 7.13, 95% CI 3.12 to 16.31; seven studies; 568 participants).Although a large number of studies reported time required for tracheal intubation (55 studies; 6249 participants), we did not present an effects estimate for this outcome owing to the extremely high level of statistical heterogeneity (I2 = 96%). AUTHORS' CONCLUSIONS Videolaryngoscopes may reduce the number of failed intubations, particularly among patients presenting with a difficult airway. They improve the glottic view and may reduce laryngeal/airway trauma. Currently, no evidence indicates that use of a VLS reduces the number of intubation attempts or the incidence of hypoxia or respiratory complications, and no evidence indicates that use of a VLS affects time required for intubation.


title: Peritoneal macrophage alterations caused by naturally occurring mouse hepatitis virus.

text: During routine harvest of murine resident peritoneal cells for macrophage function assays the authors recently noted that mice showed a 3-4-fold spontaneous increase in number of peritoneal cells within 1 week of being placed in one of their animal rooms. While the mice appeared clinically normal, the collected macrophages had highly convoluted membranes, showed enhanced spontaneous tumor cell killing, and showed increased erythrophagocytosis. Histopathologic findings included mild peritonitis and occasional foci of individual hepatocyte necrosis. The results of routine murine serologic studies and bacterial cultures of the peritoneal cavity were negative. Immunosuppressed mice placed in this room showed severe hepatic necrosis within 4 days, and ultrastructural particles characteristic of corona virus could be demonstrated in the necrotic foci. Mouse hepatitis virus (MHV) was isolated from these livers. Untreated mice showed positive MHV titers, as detected by the enzyme-linked immunoabsorbent assay (ELISA) after 21 days in the room. This episode demonstrates that MHV have profound effects on macrophage parameters while causing few clinical signs or histopathologic alterations. Secondly, the complement fixation assay for MHV as included in routine viral screens appears relatively insensitive for detecting outbreaks of MHV.


title: The ethics of sharing preliminary research findings during public health emergencies: a case study from the 2009 influenza pandemic.

text: During the 2009 A(H1N1) influenza pandemic, a suite of studies conducted in Canada showed an unexpected finding, that patients with medically attended laboratory-confirmed pandemic influenza were more likely to have received seasonal influenza vaccination than test-negative control patients. Different bodies, including scientific journals and government scientific advisory committees, reviewed the evidence simultaneously to determine its scientific validity and implications. Decision-making was complicated when the findings made their way into the media. The normal trajectory of non-urgent research includes peer-review publication after which decision-makers can process the information taking into account other evidence and logistic considerations. In the situation that arose, however, the congruence of an unexpected finding and the simultaneous review of the evidence both within and outside the traditional peer-review sphere raised several interesting issues about how to deal with emerging evidence during a public health emergency. These events are used in this article to aid discussion of the complex interrelationship between researchers, public health decision-makers and scientific journals, the trade-offs between sharing information early and maintaining the peer-review quality assurance process, and to emphasise the need for critical reflection on the practical and ethical norms that govern the way in which research is evaluated, published and communicated in public health emergencies.


California edges towards farming drug-producing rice.


title: Dissecting the requirement for subgenomic promoter sequences by RNA recombination of brome mosaic virus in vivo: evidence for functional separation of transcription and recombination.

text: Previously, we and others mapped an increased homologous recombination activity within the subgenomic promoter (sgp) region in brome mosaic virus (BMV) RNA3. In order to correlate sgp-mediated recombination and transcription, in the present work we used BMV RNA3 constructs that carried altered sgp repeats. We observed that the removal or extension of the poly(U) tract reduced or increased recombination, respectively. Deletion of the sgp core hairpin or its replacement by a different stem-loop structure inhibited recombination activity. Nucleotide substitutions at the +1 or +2 transcription initiation position reduced recombination. The sgp core alone supported only basal recombination activity. The sites of crossovers mapped to the poly(U) region and to the core hairpin. The observed effects on recombination did not parallel those observed for transcription. To explain how both activities operate within the sgp sequence, we propose a dual mechanism whereby recombination is primed at the poly(U) tract by the predetached nascent plus strand, whereas transcription is initiated de novo at the sgp core.


title: Coronavirus phylogeny based on base-base correlation.

text: With more and more complete genome sequences having been released, phylogenetic analysis is entering a new era--that of phylogenomics. In this paper, a novel phylogenomic method, named as Base-Base Correlation (BBC), has been proposed to infer phylogenetic relationships from complete genomes, with particular emphasis on coronavirus phylogeny. Following the high-profile publicity of SARS outbreaks, a renewed interest in coronavirus has been promoted and two novel human coronaviruses (NL63 and HKU1) have been identified. Coronavirus phylogenomics based on BBC is well consistent with that of previous studies. BBC, to study genome information structure based on information theory, provides a novel alignment-free phylogenomic methodology in postgenome informatics.


title: Addressing hospital nurses' fear of abandonment in a bioterrorism emergency.

text: Most hospitals' disaster plans are extensive and effective at establishing an incident command center, directing material and personnel resources, and triaging patients. However, few organizations have assessed caregivers' needs and fears related to disaster response. When nurses have been interviewed on this topic, findings indicated complex concerns involving fear of loss (e.g., loss of order in their work environment, loss of safe work conditions, loss of freedom to come and go at will, and loss of trust in their hospital's commitment to their best interest). The sobering result of anticipating these losses is fear of abandonment. The purpose of this article is to address factors identified by hospital-based nurses that contribute to their fears of abandonment in a bioterrorism emergency. Hospitals that choose to respond to these concerns will exemplify best practice toward care of the community and care of their own nurses.


title: Performance and Cost evaluation of one commercial and six in-house conventional and real-time reverse transcription-pcr assays for detection of severe acute respiratory syndrome coronavirus.

text: We evaluated seven reverse transcription-PCR (RT-PCR) assays, including six in-house assays and one commercial assay for the detection of severe acute respiratory syndrome coronavirus (SARS-CoV) RNA in clinical specimens. RT-PCR assays targeted different genomic regions and included three conventional assays (one nested and two non-nested) run on a conventional heat block and four real-time assays performed in a LightCycler (LC; Roche Diagnostics). All in-house assays were optimized for assay parameters, including MgCl2, primer, and probe concentrations. The commercial assay was the RealArt HPA CoV RT-PCR assay (Artus), which was run in the LC. Testing serial dilutions of cultured SARS-CoV showed that the analytical sensitivity of the assays ranged from 10(-8) to 10(-6), corresponding to 1 and 100 copies of viral RNA, respectively. Significant differences in analytical sensitivities were observed between assays (P < 0.01, probit regression analysis for 50% sensitivity levels for the top two assays versus the others). Testing 68 clinical specimens (including 17 respiratory tract specimens, 29 urine samples, and 22 stools or rectal swabs) demonstrated that six of the seven assays detected at least 17 of 18 positives (defined as positive in at least two assays), and two of the assays had a sensitivity of 100%. There were no significant differences in sensitivity between the assays (P = 0.5 [Cochrance Q test, least sensitive 15 of 18 versus 18 of 18]). The specificities of the assays ranged from 94.0 to 100% without significant differences (P = 0.25 to 0.5 [McNemar test]). The reagent and technologist cost of performing the in-house PCR assays ranged from $5.46 to $9.81 Canadian dollars (CDN) per test. The commercial assay cost was considerably higher at $40.37 per test. The results demonstrated good performance for all assays, providing laboratories that need to do SARS RNA testing with a choice of assay formats.