title: Three Eras in Global Tobacco Control: How Global Governance Processes Influenced Online Tobacco Control Networking.

text: Online networks can serve as a platform to diffuse policy innovations and enhance global health governance. This study focuses on how shifts in global health governance may influence related online networks. We compare social network metrics (average degree centrality [AVGD], density [D] and clustering coefficient [CC]) of Globalink, an online network of tobacco control advocates, across three eras in global tobacco control governance; pre-Framework Convention on Tobacco Control (FCTC) policy transfer (1992-1998), global regime formation through the FCTC negotiations (1999-2005), and philanthropic funding through the Bloomberg Initiative (2006-2012). Prior to 1999, Globalink was driven by a handful of high-income countries (AVGD=1.908 D=0.030, CC=0.215). The FCTC negotiations (1999-2005) corresponded with a rapid uptick in the number of countries represented within Globalink and new members were most often brought into the network through relationships with regional neighbors (AVGD=2.824, D=0.021, CC=0.253). Between 2006 and 2012, the centrality of the US in the network increases significantly (AVGD=3.414, D=0.023, CC=0.310). The findings suggest that global institutionalization through WHO, as with the FCTC, can lead to the rapid growth of decentralized online networks. Alternatively, private initiatives, such as the Bloomberg Initiative, can lead to clustering in which a single source of information gains increasing influence over an online network.


The time of cholera.


title: Synthesis of minus-strand copies of a viral transgene during viral infections of transgenic plants.

text: Viral transgenes designed to provide resistance to specific plant viruses frequently consist of the coat protein gene and a contiguous 3' untranslated region (3'UTR) of viral origin. In many RNA viruses the viral 3'UTR establishes a recognition and initiation site for viral RNA replication. Thus the transgenic transcript may contain a functional virus replication site. Experiments were designed to determine if a challenging virus would recognize this replication site on a nuclear derived transcript and synthesize the complementary RNA. These data demonstrate that upon infection by a virus that recognizes the viral replication site, a full-length complement of the transgenic transcript is produced. In these experiments the replication complex of Brome Mosaic bromovirus recognized the transgenic transcript derived from a Cowpea Chlorotic Mottle bromovirus transgene. The resulting RNA may contribute to RNA recombination events.


PreScription: COVID-19 and Compounding Pharmacists.


title: An immunodominant CD4+ T cell site on the nucleocapsid protein of murine coronavirus contributes to protection against encephalomyelitis.

text: The murine coronavirus neurotropic strain JHM (MHV-JHM) nucleocapsid (N) protein induces a strong T-helper cell response in Lewis rats. It has been shown previously that N-specific CD4+ T cells can confer protection against acute disease upon transfer to otherwise lethally infected rats. To define the major antigenic regions that elicit this T cell response, truncated fragments of N protein were expressed from a bacterial expression vector and employed as T cell antigens. Lymphocytes from either MHV-JHM-infected or immunized rats were stimulated in culture with virus antigen, grown and tested for their specificity to the N protein fragments. The carboxy-terminally located C4-N fragment (95 amino acids) induced the most pronounced proliferative response irrespective of whether the lymphocyte culture was derived from immunized or MHV-JHM-infected rats. We established T cell lines specific for the truncated N protein fragments and tested their potential to mediate protection by transfer experiments. Only the T cell line C4-N and the T cell line specific for the full-length N protein were protective. By contrast, all truncated N protein fragments elicited a humoral immune response and contained antigenic sites recognized by antibodies from diseased rats.


The reviewers were thinking wisely.


title: Analysis of age-dependent resistance to murine coronavirus JHM infection in mice.

text: Resistance to intraperitoneal murine coronavirus JHM infection in mice develops with age. C3H mice were found to be fully susceptible up to the age of 20 days and resistant after 23 days of age. Protection of susceptible animals from death due to infection could be achieved by maternal antibodies or by transfer of spleen cells from immunized, but not from nonimmunized, donor mice. Lack of protection by transfer of unprimed adult spleen cells was not related to immunosuppression by the host. Moreover, resistance of adult mice could not be abrogated by application of lymphocytes from suckling mice, although immune suppression by other means did affect the resistance of adult animals. On the other hand, spleen cells from nonimmunized mice could be primed with inactivated JHM virus in suckling mice and protected these mice from death due to a subsequent virus infection. Thus, the outcome of infection with JHM virus in suckling and adult mice can be influenced by immunological events, but is not exclusively due to the different stages of immune competence.


title: Results of Laparoscopic Sleeve Gastrectomy in 541 Consecutive Patients with Low Baseline Body Mass Index (30-35 kg/m2).

text: BACKGROUND Laparoscopic sleeve gastrectomy (LSG) is currently the leading bariatric procedure and targets, among other obesity classes, patients with BMI 30-35 kg/m2, which are reaching alarming proportions. METHODS Between February 2010 and August 2015, data on 541 consecutive patients with BMI 30-35 kg/m2 undergoing LSG were prospectively collected and analyzed. RESULTS Mean age was 32 ± 8 years (13-65) and 419 (77.4 %) were women. Preoperative weight was 92.0 ± 8.8 kg (65-121) and BMI was 32.6 ± 1.5 kg/m2 (30-35). Comorbidities were detected in 210 (39 %) patients. Operative time was 74 ± 12 min (40-110) and postoperative stay was 1.7 ± 0.22 days (1-3). There were no deaths, leaks, abscesses or strictures and the rate of hemorrhage was 1.2 %. At 1 year, 98 % were followed and BMI decreased to 24.7 ± 1.6, the percentage of total weight loss (% TWL) was 24.1 ± 4.7 while the percentage of excess BMI loss (%EBMIL) reached 106.1 ± 24.1. At 5 years, 76 % of followed patients achieved a ≥50 % EBMIL. CONCLUSION With appropriate surgical expertise, LSG in patients with BMI 30-35 kg/m2 achieved excellent outcomes with a zero fistula rate.


title: Human macrophages are susceptible to coronavirus OC43.

text: Adherent adult and cord blood macrophages were infected with human coronavirus OC43 at a multiplicity of 1-1.5 and washed twice to remove unbound virus. Virus progeny was detected in the supernatant on day 1 and peaked at 2-3 days at an average titer of 5 +/- 3.9 x 10(6) pfu/ml from seven samples. Viral RNA was detected by nested set RT-PCR in infected macrophages incubated for 48 hr. Intracellular viral nucleocapsid was detected in 15% of the cells and surface staining for viral spike antigen was observed using monoclonal antibodies. Amplification of infectious virus and detection viral RNA and antigen synthesis in macrophages in vitro indicates susceptibility to OC43 virus.


title: Enhanced virulence mediated by the murine coronavirus, mouse hepatitis virus strain JHM, is associated with a glycine at residue 310 of the spike glycoprotein.

text: The coronavirus, mouse hepatitis virus strain JHM, causes acute and chronic neurological diseases in rodents. Here we demonstrate that two closely related virus variants, both of which cause acute encephalitis in susceptible strains of mice, cause markedly different diseases if mice are protected with a suboptimal amount of an anti-JHM neutralizing antibody. One strain, JHM.SD, caused acute encephalitis, while infection with JHM.IA resulted in no acute disease. Using recombinant virus technology, we found that the differences between the two viruses mapped to the spike (S) glycoprotein and that the two S proteins differed at four amino acids. By engineering viruses that differed by only one amino acid, we identified a serine-to-glycine change at position 310 of the S protein (S310G) that recapitulated the more neurovirulent phenotype. The increased neurovirulence mediated by the virus encoding glycine at position S310 was not associated with a different tropism within the central nervous system (CNS) but was associated with increased lateral spread in the CNS, leading to significantly higher brain viral titers. In vitro studies revealed that S310G was associated with decreased S1-S2 stability and with enhanced ability to mediate infection of cells lacking the primary receptor for JHM ("receptor-independent spread"). These enhanced fusogenic properties of viruses encoding a glycine at position 310 of the S protein may contribute to spread within the CNS, a tissue in which expression of conventional JHM receptors is low.


title: [SARS vaccines].

text: Severe acute respiratory syndrome (SARS) is an emerging disease derived from wild animals and is highly pathogenic with a high mortality. A novel coronavirus SARS coronavirus was identified to be a causative agent for SARS. Since its discovery, many trials have been executed to establish the prophylactic and therapeutics toward SARS using laboratory animals. A number of different types of vaccines, such as inactivated virus vaccine, subunit vaccine, DNA vaccine and vaccine using viral expression vector as well as attenuated live vaccine so far reported indicate that neutralizing antibodies play an important role for protection. However, vaccine applicable to humans with the high efficacy and safety has not yet been reported.


title: Metro Atlanta responds to West Nile virus: a coordinated public health response.

text: Three and a half million people live in metropolitan Atlanta, in multiple counties with varying population bases, resources, issues and separate boards of health. Historically, public health issues have been managed within each county, with very little sharing of information among counties. The 1999 West Nile virus (WNV) outbreak in the Northeast caused public health officials in Atlanta to recognize the potential for the disease to spread to Georgia and the need to develop a coordinated, multi-jurisdictional response plan. This plan would need to address a new disease with little scientific data to predict how it might behave in a new environment and would also require closely coordinated communication among the local/state public health entities and elected officials. In early 2000, staff from the five health districts in the metro Atlanta area and the state health department voluntarily convened the Metro Atlanta Surveillance Task Force (MASTF) to create the Metro Atlanta West Nile Virus Response Plan. This plan utilizes a coordinated effort encompassing public education, surveillance, and mosquito control. With this plan in place, when the first human case of WNV was detected in Atlanta, the public heard consistent health messages about preventive measures to lower their risk of illness and the metro counties were able to carry out a successful uniform approach to mosquito control. This plan has received recognition by the National Association of County and City Health Officials (NACCHO) as a 2004 Model Practice, demonstrating exemplary and replicable qualities in response to a local public health need. Since the early days of the emergence of WNV in the metro Atlanta area, MASTF has continued to be a viable, evolving entity, managing and anticipating health issues. The MASTF plan is a successful effort to develop consistent policies and procedures for disease surveillance in a heavily populated area with multiple local health departments.


title: IFN-induced attrition of CD8 T cells in the presence or absence of cognate antigen during the early stages of viral infections.

text: Profound lymphopenia has been observed during many acute viral infections, and our laboratory has previously documented a type I IFN-dependent loss of CD8 T cells immediately preceding the development of the antiviral T cell response. Most memory (CD44(high)) and some naive (CD44(low)) CD8 T cells are susceptible to IFN-induced attrition, and we show in this study that the IFN-induced attrition of CD8(+)CD44(high) T cells is associated with elevated activation of caspase-3 and caspase-8. We questioned whether TCR engagement by Ag would render CD8 T cells resistant to attrition. We tested whether a high concentration of Ag (GP33 peptide) would protect lymphocytic choriomeningitis (LCMV)-specific naive CD8 T cells (TCR transgenic P14 cells specific for the GP33 epitope of LCMV) and memory CD8 T cells (GP33-specific LCMV-immune cells) from depletion. Both naive P14 and memory GP33-specific donor CD8 T cells decreased substantially 16 h after inoculation with the Toll receptor agonist and IFN inducer, poly(I:C), regardless of whether a high concentration of GP33 peptide was administered to host mice beforehand. Moreover, donor naive P14 and LCMV-specific memory cells were depleted from day 2 LCMV-infected hosts by 16 h posttransfer. These results indicate that Ag engagement does not protect CD8 T cells from the IFN-induced T cell attrition associated with viral infections. In addition, computer models indicated that early depletion of memory T cells may allow for the generation for a more diverse T cell response to infection by reducing the immunodomination caused by cross-reactive T cells.


title: A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo.

text: A deletion mutant of severe acute respiratory syndrome coronavirus (SARS-CoV) has been engineered by deleting the structural E gene in an infectious cDNA clone that was constructed as a bacterial artificial chromosome (BAC). The recombinant virus lacking the E gene (rSARS-CoV-DeltaE) was rescued in Vero E6 cells. The recovered deletion mutant grew in Vero E6, Huh-7, and CaCo-2 cells to titers 20-, 200-, and 200-fold lower than the recombinant wild-type virus, respectively, indicating that although the E protein has an effect on growth, it is not essential for virus replication. No differences in virion stability under a wide range of pH and temperature were detected between the deletion mutant and recombinant wild-type viruses. Although both viruses showed the same morphology by electron microscopy, the process of morphogenesis seemed to be less efficient with the defective virus than with the recombinant wild-type one. The rSARS-CoV-DeltaE virus replicated to titers 100- to 1,000-fold lower than the recombinant wild-type virus in the upper and lower respiratory tract of hamsters, and the lower viral load was accompanied by less inflammation in the lungs of hamsters infected with rSARS-CoV-DeltaE virus than with the recombinant wild-type virus. Therefore, the SARS-CoV that lacks the E gene is attenuated in hamsters, might be a safer research tool, and may be a good candidate for the development of a live attenuated SARS-CoV vaccine.


title: MicroRNA-Mediated Down-Regulation of Apoptosis Signal-Regulating Kinase 1 (ASK1) Attenuates the Apoptosis of Human Mesenchymal Stem Cells (MSCs) Transplanted into Infarcted Heart.

text: Stem cell therapy using adult stem cells, such as mesenchymal stem cells (MSCs) has produced some promising results in treating the damaged heart. However, the low survival rate of MSCs after transplantation is still one of the crucial factors that limit the therapeutic effect of stem cells. In the damaged heart, oxidative stress due to reactive oxygen species (ROS) production can cause the death of transplanted MSCs. Apoptosis signal-regulating kinase 1 (ASK1) has been implicated in the development of oxidative stress-related pathologic conditions. Thus, we hypothesized that down-regulation of ASK1 in human MSCs (hMSCs) might attenuate the post-transplantation death of MSCs. To test this hypothesis, we screened microRNAs (miRNAs) based on a miRNA-target prediction database and empirical data and investigated the anti-apoptotic effect of selected miRNAs on human adipose-derived stem cells (hASCs) and on rat myocardial infarction (MI) models. Our data indicated that miRNA-301a most significantly suppressed ASK1 expression in hASCs. Apoptosis-related genes were significantly down-regulated in miRNA-301a-enriched hASCs exposed to hypoxic conditions. Taken together, these data show that miRNA-mediated down-regulation of ASK1 protects MSCs during post-transplantation, leading to an increase in the efficacy of MSC-based cell therapy.


title: Psychosocial impact among the public of the severe acute respiratory syndrome epidemic in Taiwan.

text: During the 2003 outbreak, severe acute respiratory syndrome (SARS) spread to more than 30 countries. Not only did it cause severe health problems but it also imposed a great psychological impact on the public. SARS emerged in Taiwan during April 2003. This study investigates the psychosocial impact and the associated factors of depression of the SARS epidemic in Taiwan when the epidemic had just been controlled. A total of 1552 respondents were recruited in the study by random selection from the telephone book. Demographic data, SARS experience, self-perceived health state, neighborhood relationships, and depression were surveyed by telephone interviewing. Respondents were grouped as 'impacted group' and 'non-impacted group' according to whether they or their friends and family had been quarantined, or suspected of being infected. The psychosocial impact and associated factors were compared between the two groups. The 'impacted group' had higher depressive levels, poorer neighborhood relationships, poorer self-perceived health, and a higher economic impact than the 'non-impacted group'. The poorer self-perceived health and economic impact factors were associated with depression. The neighborhood relationship factor was negatively associated with depression for the 'impacted group', but not for the 'non-impacted group'. The 'impacted group' had experienced greater psychosocial impact possibly due to the SARS impact, the economic downturn, poor self-perceived health conditions, and decreased social support systems. An appropriate mental health intervention to improve the self-perceived health condition, to provide instrumental and psychological support for the 'impacted group', and to decrease the stigmatization and discrimination from the public could have buffered the psychological impact from this epidemic disaster.


title: Spatial and temporal analysis of human infection with avian influenza A(H7N9) virus in China, 2013.

text: Descriptive and geographic information system methods were used to depict the spatial and temporal characteristics of the outbreak of human infection with a novel avian influenza A(H7N9) virus in mainland China, the peak of which appeared between 28 March and 18 April 2013. As of 31 May 2013, there was a total of 131 reported human infections in China, with a cumulative mortality of 29% (38/131). The outbreak affected 10 provinces, with 106 of the cases being concentrated in the eastern coastal provinces of Zhejiang, Shanghai and Jiangsu. Statistically significant spatial clustering of cumulative human cases was identified by the Cuzick–Edwards’ k-nearest neighbour method. Three spatio-temporal clusters of cases were detected by space–time scan analysis. The principal cluster covered 18 counties in Zhejiang during 3 to 18 April (relative risk (RR): 26.39;p<0.0001), while two secondary clusters in March and April covered 21 counties along the provincial boundary between Shanghai and Jiangsu (RR: 6.35;p<0.0001) and two counties in Jiangsu (RR: 72.48;p=0.0025). The peak of the outbreak was in the eastern coastal provinces of Zhejiang, Shanghai and Jiangsu that was characterised by statistically significant spatio-temporal aggregation, with a particularly high incidence in March and April 2013.


title: 3C-like proteinase from SARS coronavirus catalyzes substrate hydrolysis by a general base mechanism.

text: SARS 3C-like proteinase has been proposed to be a key enzyme for drug design against SARS. Lack of a suitable assay has been a major hindrance for enzyme kinetic studies and a large-scale inhibitor screen for SARS 3CL proteinase. Since SARS 3CL proteinase belongs to the cysteine protease family (family C3 in clan CB) with a chymotrypsin fold, it is important to understand the catalytic mechanism of SARS 3CL proteinase to determine whether the proteolysis proceeds through a general base catalysis mechanism like chymotrypsin or an ion pair mechanism like papain. We have established a continuous colorimetric assay for SARS 3CL proteinase and applied it to study the enzyme catalytic mechanism. The proposed catalytic residues His41 and Cys145 were confirmed to be critical for catalysis by mutating to Ala, while the Cys145 to Ser mutation resulted in an active enzyme with a 40-fold lower activity. From the pH dependency of catalytic activity, the pK(a)'s for His41 and Cys145 in the wild-type enzyme were estimated to be 6.38 and 8.34, while the pK(a)'s for His41 and Ser145 in the C145S mutant were estimated to be 6.15 and 9.09, respectively. The C145S mutant has a normal isotope effect in D(2)O for general base catalysis, that is, reacts slower in D(2)O, while the wild-type enzyme shows an inverse isotope effect which may come from the lower activation enthalpy. The pK(a) values measured for the active site residues and the activity of the C145S mutant are consistent with a general base catalysis mechanism and cannot be explained by a thiolate-imidazolium ion pair model.


title: Getting more from less: algorithms for rapid protein identification with multiple short peptide sequences.

text: We describe two novel sequence similarity search algorithms, FASTS and FASTF, that use multiple short peptide sequences to identify homologous sequences in protein or DNA databases. FASTS searches with peptide sequences of unknown order, as obtained by mass spectrometry-based sequencing, evaluating all possible arrangements of the peptides. FASTF searches with mixed peptide sequences, as generated by Edman sequencing of unseparated mixtures of peptides. FASTF deconvolutes the mixture, using a greedy heuristic that allows rapid identification of high scoring alignments while reducing the total number of explored alternatives. Both algorithms use the heuristic FASTA comparison strategy to accelerate the search but use alignment probability, rather than similarity score, as the criterion for alignment optimality. Statistical estimates are calculated using an empirical correction to a theoretical probability. These calculated estimates were accurate within a factor of 10 for FASTS and 1000 for FASTF on our test dataset. FASTS requires only 15-20 total residues in three or four peptides to robustly identify homologues sharing 50% or greater protein sequence identity. FASTF requires about 25% more sequence data than FASTS for equivalent sensitivity, but additional sequence data are usually available from mixed Edman experiments. Thus, both algorithms can identify homologues that diverged 100 to 500 million years ago, allowing proteomic identification from organisms whose genomes have not been sequenced.


title: Acute Respiratory Distress Syndrome Secondary to Influenza A(H1N1)pdm09: Clinical Characteristics and Mortality Predictors.

text: BACKGROUND Acute respiratory distress syndrome secondary to influenza A(H1N1)pdm09 virus is the leading cause of death among this patient population. Expanding the knowledge of its course and predictors of mortality is relevant to decision making. We aimed to describe the clinical characteristics and identify factors associated with mortality in patients with acute respiratory distress syndrome secondary to influenza A(H1N1)pdm09 during the 2013-2014 influenza season. METHODS This is an observational study of a prospective cohort of 70 patients with acute respiratory distress syndrome and influenza A(H1N1) pdm09 seen in an academic medical center. Multivariate logistic regression was used to identify the independent mortality predictors. Bootstrap was used for internal model validation. RESULTS This cohort was represented by young adults (43 ± 11 years old). Obesity was present in 62.5% and was not associated with mortality. Mortality at 28 days and at discharge from the respiratory intensive care unit was 14 and 20%, respectively. All patients met the criteria for acute respiratory distress syndrome, 73% had vasodilatory shock, and 27.1% had acute kidney injury on respiratory intensive care unit admission. We observed a high incidence of intensive care unit-acquired weakness (81.4%). Ventilator-associated pneumonia developed in 47.1% and was not associated with mortality. In multivariate analysis, independent risk factors for intensive care unit mortality were age (odds ratio [OR] = 1.102), white blood cell count (OR = 1.22), and lactate dehydrogenase levels (OR = 1.004) on admission to the intensive care unit. CONCLUSIONS We described the clinical characteristics and course of a cohort of patients with acute respiratory distress syndrome secondary to influenza A(H1N1)pdm09, and developed a predictive model of mortality based on the covariates age, levels of lactate dehydrogenase, and white cell count on admission to the respiratory intensive care unit.


title: Retrieval of distorted pipeline embolic device using snare-loop.

text: BACKGROUND Flow-diverter stents are increasingly being used in the endovascular treatment of intracranial aneurysms. Because of this increased usage, the occurrence of stent malpositioning, intra-arterial distortion, or migration will likely increase as well. METHODS We describe the conformational twisting and deformity of a pipeline embolic device (PED) stent (Covidien, EV3) within the carotid artery during stent placement, with the subsequent immediate removal of the stent by using an endovascular snare-loop to successfully remove the device. RESULTS The PED can be successfully removed using a snare-loop device when indicated. In this case, following removal of the initial PED, the aneurysms can be successfully treated with the placement of a second PED. CONCLUSION It is critical to have a contingency plan for the removal of malpositioned or otherwise deformed stents that could potentially represent significant sources of iatrogenic stroke.


Global health science: a threat and an opportunity for collaborative clinical science.


Opportunistic Infections in Research Rodents: The Challenges Are Great and the Hour is Late.


title: Receptor-dependent coronavirus infection of dendritic cells.

text: In several mammalian species, including humans, coronavirus infection can modulate the host immune response. We show a potential role of dendritic cells (DC) in murine coronavirus-induced immune modulation and pathogenesis by demonstrating that the JAW SII DC line and primary DC from BALB/c mice and p/p mice with reduced expression of the murine coronavirus receptor, murine CEACAM1a, are susceptible to murine coronavirus infection by a receptor-dependent pathway.


title: Primary Stenting for Acute Ischemic Stroke Using the Enterprise Intracranial Stent: 2-Year Results of a Phase-I Trial.

text: BACKGROUND The preliminary results of a prospective consecutive series of 20 patients who underwent Enterprise-assisted recanalization for acute ischemic stroke were recently reported. Recanalization to thrombolysis in myocardial infarction (TIMI) grade 2 (n = 6) or 3 (n = 12) flow was achieved in 18 patients (90% revascularization rate). Good outcome (modified Rankin Scale [mRS] score of ≤2) was obtained in 10 patients (50%) at 30 days. Here, we report the 2-year clinical follow-up data for patients enrolled in that prospective study. METHODS Study patients were scheduled for examinations 2 years postprocedure at which time mRS and Barthel indices were obtained. RESULTS Among 12 survivors at 2 years, 11 of the 20 (55%) study patients improved to mRS score ≤2 and 1 (5%) patient was disabled with an mRS 4. Of the 11 patients with mRS 0-2 scores, 10 patients had a Barthel index of 100, and the 11th had a Barthel index of 95. One patient improved from mRS 3 to 2 during the interval between the 6- and 12-month postintervention evaluations after intervention. Eight of 13 (62%) survivors underwent follow-up imaging at 6 months without evidence of instent stenosis or thrombosis. CONCLUSION At 2 years of follow-up, improvement in quality of life after acute stroke intervention was sustained; and 11 of 12 (92%) survivors had an excellent functional outcome. Improvement in functional status can occur even up to 1 year after stroke intervention. These results 2 years after acute stroke intervention demonstrate sustained benefit from acute intervention. ABBREVIATIONS AISacute ischemic strokeCTcomputed tomographicFDAFood and Drug AdministrationIVintravenousMCAmiddle cerebral arterymRSmodified Rankin ScaleNIHSSNational Institutes of Health Stroke Scale ScoreSWIFTSolitaire FR With the Intention For Thrombectomy (SWIFT)TIMIthrombolysis in myocardial infarctiontPAtissue plasminogen activatorTREVOThrombectomy REvascularization of large Vessel Occlusions.


Vivax Malaria: Benign No More.


title: Dot-blot hybridization using digoxigenin-labeled cDNA probe complementary to the S1 gene of avian infectious bronchitis virus permits discrimination between virus strains.

text: Digoxigenin-dUTP-labeled DNA probe was prepared from a cDNA clone complementary to the gene encoding S1 region of the spike protein of infectious bronchitis coronavirus (IBV) strain M41. The probe exclusively reacted with four strains at 56 degrees C which were grouped to the same serotype as the strain used for the probe. In contrast, at 68 degrees C, the probe reacted only with the homologous strain and did not react even with the strains belonging to the same serotype. The dot-blot hybridization thus appeared serotype-specific at 56 degrees C and strain-specific at 68 degrees C. In addition, it was revealed that the S1 gene has some nucleotide sequence variation even among strains in the same serotype. This technique should be applied to determining serotypes of the virus isolates and to differentiating field isolates from the vaccine strain.


title: [Epidemiology of viral nosocomial infections in pediatrics].

text: Nosocomial viral infections account for at least 5% of the total of NI and reach 23% in pediatric wards. The nosocomial infection (NI) incidence rate varies from 0.59 to 0.72 per 100 patients in pediatric wards. Many viruses have been associated with NI in pediatric wards. Rotavirus and respiratory syncytial virus (RSV) are the most frequent. Other viruses frequently identified are: calicivirus, adenovirus, astrovirus, influenza et para-influenza, rhinovirus and coronavirus. Asymptomatic infections occur frequently. The period of communicability varies and depends on the virus. It often begins before the clinical signs appear and ends after the healing. Viral shedding may be intermittent. Children and hospital environment and less frequently hospital staff are the main source for the virus. Poor handwashing results in direct spread to patient or self-inoculation even for respiratory viruses like RSV and rhinovirus. The main risk factors for NI are prolonged hospital stay, past history of prematurity and low age. Immunocompromised patients constitute a special high-risk group. Understaffing is also a risk factor. Minimal infective doses depend on the route of inoculation and the kind of virus. Low doses are for example sufficient for rotavirus, adenovirus and calicivirus. Viral inactivation is all the more easy when there is an envelope. Handwashing and appropriate isolation (technical and geographical) are the mainstay of prevention of viral NI. Vaccines are promising, especially for rotavirus.


title: International comparisons in critical care: a necessity and challenge.

text: PURPOSE OF REVIEW Understanding variation in critical care resources, and delivery of care between countries will allow for improved disaster planning, evaluation of research findings, and assessment of the utility of critical care itself. This review describes the available data for international comparisons and the many factors that need to be addressed for an appropriate interpretation of results. RECENT FINDINGS Recent studies on subgroups of critical care patients include data from many different countries. These new studies provide important information on the overall incidence of these disease states, but most of these international studies do not take into account the critical care resources of the countries being discussed. For an appropriate interpretation of findings the relevant baseline critical care resources, prevalence of diseases, and cultural practices, need to be quantified. The existence of these other factors prevents the use of a severity of illness scoring system alone to account for differences between countries. SUMMARY Many recent critical care studies include data from multiple countries. With continued movement towards international studies, and improvements in data collection systems, comparisons between countries are becoming easier. These findings need to be interpreted in the context of all the relevant country information.


title: [Expression of the monoclonal antibody against nucleocapsid antigen of SARS-associated coronavirus in autopsy tissues from SARS patients].

text: OBJECTIVE To investigate the presence and distribution of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) in autopsy tissues obtained from patients died of SARS. METHODS Immunohistochemical technique was applied in 4 fatal SARS cases to examine the autopsy tissues including the lungs, spleen, lymph nodes, brain, pituitary, heart, liver, kidney, pancreas, trachea, esophagus, gastrointestinal tract, adrenal glands, parathyroids, skin and bone marrow. RESULTS Immunohistochemistry identified positive monoclonal antibody against SARS-CoV nuceeocapsid (N) protein in the alveolar epithelium and the infiltrating monocytes or macrophages in the lung, spleen and lymph nodes; the presence of the antibody was also detected in the serous gland epithelium of the trachea/bronchus, squamous epithelium of the esophagus, the gastric parietal cells, the epithelium of the intestinal tract, acidophilic cells in the parathyroids and pituitary, acinus cells in the pancreas, adrenal cortical cells, sweat gland cells, small vessel endothelium, bone marrow promyelocytes, epithelial cells of the distal convoluted tubule of the kidney, brain neurons, and the hepatocytes near the central vein. CONCLUSIONS A variety of organs and tissues can be infected by SARS-CoV, and the positive expression of SARS-CoV N protein in the epithelial cells of the gastrointestinal tract, the distal convoluted tubule of the kidney and the sweat gland cells is significant for studying the transmission routes of SARS.


title: ['Severe acute respiratory syndrome' (SARS): epidemiology, clinical signs, diagnosis and prevention].

text: Severe acute respiratory syndrome (SARS) is caused by a recently identified Coronavirus (SARS-CoV). The clinical symptoms are non-specific and during the first few days in particular, are not clinically distinguishable from those of many other viral or bacterial infections. The majority of infected patients develop pneumonia within a week of the first symptoms appearing. Since November 2002 the virus has spread from South China to almost 30 other countries, where about 8500 infected individuals have been registered; about 800 people have already died from the disease (9.5%). The number of infected persons includes a noticeably high percentage of health workers. This fact underlines the importance of good infection prevention measures for each patient contact. The implementation of hygienic measures requires attention, because the infection of personnel in Toronto hospitals still occurred after the virus and transmission routes were known. It appears that transmission can be prevented with relatively simple precautions, as long as these are consistently implemented. Early recognition and isolation of a possible source are an essential part of this. SARS is a group A notifiable disease (report if suspected). In the Netherlands the general practitioner has a prominent role in assessing and treating individuals who are infected or might be infected with SARS-CoV. A protocol and a detailed action plan are available. In addition to this hospitals should be prepared for the initial reception of a patient with SARS, who presents directly to the outpatients' clinic or Casualty Department.


title: Prevention and control of emerging infections: a challenge for the 3rd millennium.

text: In the last 30 years, several emerging infections due to novel viruses have been identified, from haemorrhagic fever viruses to HIV, from the SARS-Coronavirus to Avian influenza viruses. Ecological and genetic changes are important determinants of the emergence of new viral infections, driving to an increase of R0 (the basic reproductive number) through increasing the probability of transmission. The current H5N1 epidemic may be considered a prepandemic paradigm that needs thorough investigation.


title: Rapid diagnosis by electron microscopy of avian coronavirus infection.

text: The election microscopic examination of allantoic fluid from embryonated hens' eggs inoculated with tissue homogenates from organs of birds suffering with infectious bronchitis, reveals the presence of coronaviruses as early as the first or the second passage. This method permits a rapid diagnosis and is as accurate as the standard technique.


title: Non-technical skills in minimally invasive surgery teams: a systematic review.

text: BACKGROUND Root cause analyses show that up to 70 % of adverse events are caused by human error. Strong non-technical skills (NTS) can prevent or reduce these errors, considerable numbers of which occur in the operating theatre. Minimally invasive surgery (MIS) requires manipulation of more complex equipment than open procedures, likely requiring a different set of NTS for each kind of team. The aims of this study were to identify the MIS teams' key NTS and investigate the effect of training and assessment of NTS on MIS teams. METHODS The databases of PubMed, Cochrane Library, Embase, PsycINFO, and Scopus were systematically searched according to Preferred Reporting Item for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Articles containing outcome measures related to MIS teams' key NTS, training, or assessment of NTS were included. RESULTS The search yielded 1984 articles, 11 of which were included. All were observational studies without blinding, and they differed in aims, types of evaluation, and outcomes. Only two studies evaluated patient outcomes other than operative time, and overall, the studies' quality of evidence was low. Different communication types were encountered in MIS compared to open surgery, mainly due to equipment- and patient-related challenges. Fixed teams improved teamwork and safety levels, while deficient planning and poor teamwork were found to obstruct workflow and increase errors. Training NTS mitigates these issues and improves staff attitudes towards NTS. CONCLUSIONS MIS teams' NTS are important for workflow and prevention of errors and can be enhanced by working in fixed teams. In the technological complex sphere of MIS, communication revolves around equipment- and patient-related topics, much more so than in open surgery. In all, only a few heterogeneous-design studies have examined this. In the future, the focus should shift to systematically identifying key NTS and developing effective, evidence-based team training programmes in MIS.


title: Knowledge, attitude and practice towards SARS.

text: The objective of the study was to explore the knowledge, attitude and practice of the population of Qatar towards severe acute respiratory syndrome (SARS). The study took the form of a cross-sectional community-based survey in Doha International Airport and primary health care centres situated in urban and semi-urban areas of Qatar. The survey was conducted among travellers arriving at Doha International Airport and community residents visiting health centres from April to September 2003. A total of 1,800 subjects were approached for the study of which 1,386 (77.0%) responded to the questionnaire. The study showed that although 79.4% of the total subjects claimed to know about SARS, only 8.0% had accurate knowledge about all of the symptoms of SARS. The respondents' satisfaction with the preventive measures taken by the health authorities was only 66.6%. Overall, 73.0% of the total subjects knew that SARS was highly infectious; 69.9% knew that it could spread through close contact; 67.4% were aware that high fever was the early symptom; and 60.8% thought SARS could be cured. Half of the subjects said they had gained most of their knowledge on SARS through television and radio programmes (52.2%). Three-fifths (60.1%) were afraid to travel for fear of being affected by SARS. The level of awareness of SARS among Qataris was poor (31.7%) compared to that among non-Qataris (68.3%). The level of knowledge about SARS was better among educated subjects. The survey found a high level of stress and fear among the respondents. Health authorities should be on the alert and re-examine their capacity to manage an outbreak like SARS in the event that such a mishap should recur in the future.


title: Infectious diarrhea in developed and developing countries.

text: Diarrhea from gastrointestinal infection remains a common problem. In industrialized countries, management is aimed at reducing morbidity and defining groups that may benefit from further investigation. Most infectious diarrhea is self-limiting and only requires supportive management. Viral agents are increasingly recognized as causative agents of epidemic and sporadic diarrhea. In developing countries, diarrhea is a major cause of mortality in children. Oral rehydration therapy, guided by a clinical assessment of the degree of dehydration, is cheap, simple, and effective and remains the mainstay of management of infant diarrhea. Controversies focus on the optimal formulation of oral rehydration solution. A vaccine against rotavirus has the potential to save millions of lives worldwide.


title: Proteolytic cleavage of the murine coronavirus surface glycoprotein is not required for its fusion activity.

text: The surface glycoprotein (S) of the murine hepatitis coronavirus MHV normally undergoes proteolytic cleavage during transport to the cell surface. To determine whether the cleavage of the MHV-JHM S glycoprotein is required to activate its ability to fuse cellular membranes, the protease recognition sequence in a cDNA copy of the S gene was altered from Arg-Arg-Ala-Arg-Arg into Ser-Val-Ser-Gly-Gly by site directed mutagenesis. The mutated and wild type S genes were expressed by means of recombinant vaccinia viruses and it could be shown that the mutated S protein was not cleaved when it was expressed in mouse DBT cells, in contrast to the wild type S protein. Nevertheless, the non-cleaved S protein induced extensive syncytium formation in mouse DBT cells. These results clearly indicate that the non-cleaved form of the MHV S protein is able to mediate cell membrane fusion. Thus, proteolytic cleavage is not an absolute requirement for its fusion function.


title: High affinity interaction between nucleocapsid protein and leader/intergenic sequence of mouse hepatitis virus RNA.

text: The nucleocapsid (N) protein of mouse hepatitis virus (MHV) is the major virion structural protein. It associates with both viral genomic RNA and subgenomic mRNAs and has structural and non-structural roles in replication including viral RNA-dependent RNA transcription, genome replication, encapsidation and translation. These processes all involve RNA-protein interactions between the N protein and viral RNAs. To better understand the RNA-binding properties of this multifunctional protein, the N protein was expressed in Escherichia coli as a chimeric protein fused to glutathione-S-transferase (GST). Biochemical analyses of RNA-binding properties were performed on full-length and partial N protein segments to define the RNA-binding domain. The full-length N protein and the GST-N protein fusion product had similar binding activities with a dissociation constant (K(d)) of 14 nM when the MHV 5'-leader sequence was used as ligand. The smallest N protein fragment which retained RNA-binding activity was a 55 aa segment containing residues 177-231 which bound viral RNA with a K(d) of 32 nM. A consensus viral sequence recognized by the N protein was inferred from these studies; AAUCYAAAC was identified to be the potential minimum ligand for the N protein. Although the core UCYAA sequence is often tandemly repeated in viral genomes, ligands containing one or more repeats of UCYAA showed no difference in binding to the N protein. Together these data demonstrate a high-affinity, specific interaction between the N protein and a conserved RNA sequence present at the 5'-ends of MHV mRNA.


Australian Senate votes to allow therapeutic cloning.


title: Automated Analysis of Public Health Laboratory Test Results.

text: This study investigates the use of machine learning methods for classifying and extracting structured information from laboratory reports stored as semi-structured point-form English text. This is a novel data format that has not been evaluated in conjunction with machine learning classifiers in previous literature. Our classifiers achieve human-level predictive accuracy on the binary Test Performed and 4-class Test Outcome labels. We consider symbolic approaches for predicting the highly multi-class Organism Genus and Organism Species labels. Results are discussed from the viewpoint of interpretability and generalizability to new incoming laboratory reports. Code has been made public at https://github.com/enchainingrealm/UbcDssgBccdc-Research/tree/master/src.


Is reporting many cases of COVID-19 in Iran due to strength or weakness of Iran's health system?


title: Laparoscopic mesh fixation using laser-assisted tissue soldering in a porcine model.

text: BACKGROUND AND OBJECTIVE Animal studies using open surgical models indicate that collagen solder is capable of fixation of surgical meshes without interfering with tissue integration, increasing adhesions, or increasing inflammation intraperitoneally. This study describes development of instrumentation and techniques for laparoscopic herniorrhaphy using laser-assisted soldering technology. STUDY DESIGN AND METHODS Anesthetized 20 kg to 25 kg female Yorkshire pigs underwent laparoscopy with a 3-trocar technique. Parietex TET, Parietex TEC, and Prolene mesh segments (5 x 5 cm) were embedded in 55% collagen solder. Segments were inserted by using a specially designed introducer and affixed to the peritoneum by using prototype laser devices (1.45 micro, 4.5 W continuous wave, 5-mm spot, 55 degrees C set temperature) and a custom laparoscopic handpiece (IPOM). Parietex PCO mesh was inserted and affixed using the Endo-hernia stapler (Control). Animals were recovered and underwent second-look laparoscopy at 6 weeks. Mesh sites were harvested after animals were euthanized. RESULTS The mesh-solder constructs were easily inserted and affixed in an IPOM approach. Prolene mesh tended to curl at its edges as the solder was melted. Postoperative healing was similar to that in Control segments in all cases. DISCUSSION AND CONCLUSION Collagen-based tissue soldering permits normal wound healing and may mitigate or reduce the use of staples or other foreign bodies for laparoscopic mesh fixation, prevent tissue ischemia and possibly nerve entrapment, which result in severe postoperative pain and morbidity. Laser-assisted mesh fixation is a promising alternative for laparoscopic herniorrhaphy. Further development of this strategy is warranted.


The Association of Physicians of Great Britain and Ireland 1992.


title: Estimates of the reproduction numbers of Spanish influenza using morbidity data.

text: BACKGROUND There have been several studies of the transmissibility of the 1918 (Spanish) influenza virus, which has attributed to >20 million deaths. Many of the analyses to date have involved fitting predictions from a transmission model to the observed epidemic curves from different settings. METHODS Using morbidity data from cities in Europe and America and from confined settings during the 1918 influenza pandemic, we contrast the use of several different methods based on the growth rate and final size of the epidemic, which do not rely on transmission models, to estimate the effective and basic reproduction numbers. RESULTS The effective reproduction number (the average number of secondary infectious cases produced by a typical infectious case in a given population) for the 1918 influenza virus was in the range 1.2-3.0 and 2.1-7.5 for community-based and confined settings, respectively. CONCLUSIONS Assuming further that 30 and 50% of individuals were immune to Spanish influenza after the wave in April 1918 and the first subsequent wave, respectively, these findings imply that, in a totally susceptible population, an infectious case could have led to 2.4-4.3 and 2.6-10.6 cases in community-based and confined settings, respectively. These findings for community-based populations confirm the relatively low transmissibility of the 1918 (Spanish) influenza virus, which has been found by other studies using alternative data sources and methods.


title: Patient Age and the Outcomes after Decompressive Hemicraniectomy for Stroke: A Nationwide Inpatient Sample Analysis.

text: BACKGROUND Decompressive hemicraniectomy (DHC) for space-occupying cerebral infarction in older adults remains controversial, and there are limited nationwide data evaluating the outcomes after craniectomy for stroke by patient age. METHODS Patients who underwent DHC for ischemic stroke were extracted from the Nationwide Inpatient Sample (2002-2011). Multivariable logistic regression examined in-hospital mortality and a poor outcome (death, tracheostomy and gastrostomy, or discharge to institutional care). Covariates included year of admission, comorbidities, severity indices, and treatment variables (including the timing of decompression). RESULTS Craniectomy was performed in 1673 patients: 62.4 % were aged 18-60 years, 20.6 % aged 61-70 years, and 17.0 % aged greater than 70 years. DHC was associated with reduced adjusted odds of in-hospital death compared with medical treatment alone among patients with cerebral edema in all age categories, including those older than 70 years (p ≤ 0.008). However, among surgical patients, the adjusted odds of mortality were significantly greater for patients aged 61-70 (30.7 %, p = 0.02) and greater than 70 years (34.5 %, p = 0.02), but not different for patients aged 51-60 (22.8 %), compared to those aged 18-50 years (19.7 %). The adjusted odds of a poor outcome also increased significantly with age, particularly for patients greater than 60 years. CONCLUSION In this nationwide analysis, DHC was associated with reduced mortality regardless of patient age, including among those aged greater than 70 years. However, patients aged greater than 60 years treated surgically experienced higher odds of mortality (32.4 %), discharge to institutional care (47.1 %), and a poor outcome (77.0 %) compared with younger patients.


title: Pancreatic adenocarcinoma, version 2.2014: featured updates to the NCCN guidelines.

text: The NCCN Guidelines for Pancreatic Adenocarcinoma discuss the diagnosis and management of adenocarcinomas of the exocrine pancreas and are intended to assist with clinical decision-making. These NCCN Guidelines Insights summarize major discussion points from the 2014 NCCN Pancreatic Adenocarcinoma Panel meeting. The panel discussion focused mainly on the management of borderline resectable and locally advanced disease. In particular, the panel discussed the definition of borderline resectable disease, role of neoadjuvant therapy in borderline disease, role of chemoradiation in locally advanced disease, and potential role of newer, more active chemotherapy regimens in both settings.


title: Malnutrition and infection: an update.

text: The original Scrimshaw, Taylor and Gordon conceptual framework for the interaction of nutrition and infection has well served the scientific community for almost half a century. At its core is the notion of synergistic (mutually reinforcing) and antagonistic (mutually nullifying) influences of the malnourished state on infectious conditions and vice versa. Research on a series of advancing fronts, however, has allowed the incorporation of both relevant public health issues (parasitosis, emerging infectious diseases, obesity and overweight, etc.) and advancing science (molecular immunology, oxidation biology, multiple micronutrient deficiencies, etc.). The present review is an interpretative update on close to 50 years of demographic and epidemiological evolution in the field of human nutrition and the implications for the interaction in the context of microbiological and immunological developments on the infectious side of the dialectic.


title: [The effect of central anatomical single-bundle versus anatomical double-bundle reconstruction of the anterior cruciate ligament on knee stability. a clinical study].

text: PURPOSE OF THE STUDY A comparison of the efficacy of central anatomical single-bundle (CASB) reconstruction with that of double-bundle (DB) repair of the anterior cruciate ligament (ACL) in relation to knee stability in anteroposterior translation (APT), internal rotation (IR) and external rotation (ER) of the joint. MATERIAL AND METHODS A total of 40 patients were evaluated; 20 had ACL reconstruction by the CASB technique using hamstrings and 20 underwent DB repair surgery. The average age was 31.3 years, and the group included 22 men and 18 women with 19 right and 21 left knees. The KT-1000 test was used to assess the amount of APT in the knee and rotational deviations were measured by the Rolimeter. In the DB patients, measurements were performed before surgery (on joints with ACL injury), then after reconstruction of the anteromedial (AM) or the posterolateral (PL) bundle and subsequently after repair of both ACL bundles. The CASB patients were assessed before and after graft insertion. RESULTS The average APT value was 18.5 mm for the pre-operative knees and it fell to 8.9 mm after AM bundle reconstruction. However, when the PL bundle was inserted in the first place, the average APT value was 13.1 mm only. The average values recorded after the DB and CASB reconstructions were 6.1 mm and 9.1 mm, respectively. The average IR range of motion in the pre-operative joints was 18.6 degrees. After AM bundle reconstruction it was 13.9 degrees and after PL bundle repair it was 15.3 degrees. In DB reconstruction the average IR value achieved 10.4 degrees, and in CASB repair surgery it was 13.7 degrees. The average ER range of motion in the pre-operative joints was 17.8 degrees. After AM bundle reconstruction it was 14.5 degrees and after PL bundle repair it was 14.9 degrees. In DB reconstruction the average ER value achieved 11.4 degrees, and in CASB repair surgery it was 14.5 degrees. DISCUSSION Rotational stability of the knee after ACL reconstruction is one of the most important factors in restoring physiological kinematics of the joint after ACL injury. Since there are not many studies comparing knee rotational stability after CASB with that after DB reconstructions, the results presented here may contribute to selecting the optimal method of ACL reconstruction. CONCLUSIONS The results show that, in ACL reconstruction, the DB technique provides better stability to the knee, in both APT and rotation, than the CASB method. The latter has the same effect on knee stability as the presence of the AM bundle alone. When the PL bundle is added, knee stability, in both APT and internal/external rotation, is increased in comparison with central single-bundle ACL repair. Key words:anterior cruciate ligament, navigation, central anatomical single-bundle reconstruction, double-bundle reconstruction.


title: What next in necrotizing enterocolitis?

text: Necrotizing enterocolitis (NEC) is a devastating disease of premature infants, with a mortality rate of 10-50%. It is uncommon in term infants and in premature infants who have not yet been fed. Most commonly NEC develops suddenly in a preterm infant who was otherwise well, with initial symptoms of abdominal distention, bilious or bloody emesis or gastric aspirates, hematochezia, and pneumatosis intestinalis, and sometimes progresses quickly to include bowel perforation, acidosis, shock, and death. Trigger factors (i.e. perinatal hypoxia, mild infection or formula feeding) cause focal mild intestinal mucosal injury. In the presence of proliferation of commensal bacteria, local breakdown of mucosal barrier may cause entry of bacterial products (e.g. lipopolysaccharides, platelet-activating factor). Endothelial platelet-activating factor and/or tumor necrotizing factor and/or direct stimulating effects of polymorphonuclear leukocytes cause proinflammatory cascade and focal necrosis, which increase the entry of large amounts of bacterial toxins, and then severe NEC, sepsis, and shock develop. Therapies for the prevention of NEC that appear to have some benefit are breastfeeding and antenatal steroids, and probably probiotics. Enteral immunoglobulin, polyunsaturated fatty acids, and arginine or glutamine supplementation are therapies for the prevention of NEC that do not appear to be of benefit. Enteral erythropoietin and enteral granulocyte colony-stimulating factor are promising novel therapies. Treatment options are limited to gut rest, parenteral nutrition, broad-spectrum antibiotics, and surgical interventions for enteral perforation. Two commonly used methods for NEC with intestinal perforation are laparotomy or primary peritoneal drainage ("patch, drain and wait"); however, the preferred method is controversial.


title: Diffuse alveolar lesion in BALB/c mice induced with human reovirus BYD1 strain and its potential relation with SARS.

text: The objective of this study was to investigate the pathogenicity and associated lesions of a new reovirus (ReoV) isolated from patients with Severe Acute Respiratory Syndrome (SARS) in China. Twenty-five four-week-old BALB/c female mice inoculated intranasally with either ReoV (strain BYD1) alone, or ReoV combined with SARS-CoV (strain BJF) displayed ejecting fur and loss of body weight compared with control animals. ReoV and SARS-CoV were isolated from most postmortem tissues. The histopathological features of ReoV infected animals consisted of diffuse alveolar damage, with scattered hemorrhage, hyaline membrane formation and interstitial pneumonia. A typical type II pneumocyte hyperplasia and fibrogranulomatous tissue formation in the alveolar septae were observed both in the animals inoculated simultaneously with these two viruses and in the animals inoculated firstly with SARS-CoV, followed by ReoV. The animals inoculated firstly with ReoV, followed with SARS-CoV displayed scattered hemorrhage in the alveolar septa. Furthermore, other lesions in above two combination groups included depletion of lymphocytes in the germinal center of lymph nodes in the lung hilus and the spleen, hemorrhagic necrosis in white pulp of spleen, hydroid degeneration, and fatty degeneration in the liver and kidney. Mice induced with SARS-CoV alone did not display clinical signs, characteristically hyaline membrane formation, hemorrhage and early pulmonary fibrosis in lung tissue. This study demonstrated that the newly isolated ReoV might be a virulent pathogen for BALB/c mice. Mice infected firstly with SARS-CoV, followed with ReoV developed a typical diffuse alveolar lesion.


title: S1 gene characteristics and efficacy of vaccination against infectious bronchitis virus field isolates from the United States and Israel (1996 to 2000).

text: The S1 genes of isolates of avian coronavirus infectious bronchitis virus (IBV) from commercial chickens in the US and Israel (20 isolates from each country) were studied using reverse transcription-polymerase chain reaction restriction fragment length polymorphism and sequencing. Partial sequences spanning the amino terminus region of S1 from amino acid residues 48 to 219, based on the Beaudette strain, were used for analysis. Phylogenetic clustering and high-sequence identity values were used to identify isolates that appeared to be derived from live IBV vaccines used in the two countries. Novel variant strains, unrelated by S1 sequencing and restriction fragment length polymorphism analyses to reference and vaccine strains, were also identified. Based on S1 sequence identity to available vaccines, the potential to use vaccination to control IBV infections was evaluated. Vaccination with commercial live strains Massachusetts (Mass), Arkansas (Ark) or DE/072/92, generally produced immunity against vaccine-related field isolates displaying high S1 sequence similarities (> or = 90%) to the respective vaccine strains. Immunization with a bivalent vaccine containing the Mass and Ark strains provided good cross-protection, averaging 81% against challenge with five variant isolates from the US having amino acid identity values ranging from 62 to 69% to Mass and from 68 to 83% to Ark, respectively. In contrast, the H120 vaccine strain induced low levels of protection, ranging from 25 to 58% against variant field isolates from Israel with amino acid identity values from 65 to 67%.


title: Transmission of viral respiratory infections in the home.

text: Respiratory viruses in the home exploit multiple modes of transmission. RSV is transmitted primarily by contact with ill children and contaminated objects in the environment. Influenza appears to be spread mainly by airborne droplet nuclei. Despite many years of study, from the plains of Salisbury, to the hills of Virginia, to the collegiate environment of Madison, WI, the precise routes rhinovirus takes to inflict the misery of the common cold on a susceptible population remain controversial.


title: Serological, colostral and milk responses of cows vaccinated with a single dose of a combined vaccine against rotavirus, coronavirus and Escherichia coli F5 (K99).

text: Twenty-five Ayrshire/Friesian cows were vaccinated once with a new combined vaccine against rotavirus, coronavirus and Escherichia coli F5 (K99) or given a saline placebo 31 days before the first expected calving date. Blood samples were taken from the cows at intervals from vaccination until seven days after calving and from their calves up to 28 days after birth, and colostrum and milk samples were collected from the cows at intervals for 28 days after calving. There was a significant increase in the mean specific antibody titre against all three antigens in the serum of the vaccinated animals (even in the presence of pre-existing antibody) which was accompanied by increased levels of protective antibodies to rotavirus, coronavirus and E coli F5 (K99) in their colostrum and milk for at least 28 days.


title: Current status of veterinary vaccines.

text: The major goals of veterinary vaccines are to improve the health and welfare of companion animals, increase production of livestock in a cost-effective manner, and prevent animal-to-human transmission from both domestic animals and wildlife. These diverse aims have led to different approaches to the development of veterinary vaccines from crude but effective whole-pathogen preparations to molecularly defined subunit vaccines, genetically engineered organisms or chimeras, vectored antigen formulations, and naked DNA injections. The final successful outcome of vaccine research and development is the generation of a product that will be available in the marketplace or that will be used in the field to achieve desired outcomes. As detailed in this review, successful veterinary vaccines have been produced against viral, bacterial, protozoal, and multicellular pathogens, which in many ways have led the field in the application and adaptation of novel technologies. These veterinary vaccines have had, and continue to have, a major impact not only on animal health and production but also on human health through increasing safe food supplies and preventing animal-to-human transmission of infectious diseases. The continued interaction between animals and human researchers and health professionals will be of major importance for adapting new technologies, providing animal models of disease, and confronting new and emerging infectious diseases.


title: Limitations of current prophylaxis against influenza virus infection.

text: Avian influenza has been a source of worldwide concern since Hong Kong authorities detected the first outbreak in 1997. Mainly as a result of poultry-to-human transmission, more than 200 cases of infection in humans have been attributed to the A/H5, A/H7, and A/H9 viral subtypes, with a case fatality rate for A/H5N1 infections exceeding 50%. A mutant or reassortant virus capable of efficient human-to-human transmission can set off a pandemic. Increased attention to prophylaxis against viral infection has identified several potentially complementary approaches: nonpharmacologic measures (eg, travel restrictions), vaccination, chemotherapeutic agents, and herbal/natural products. All have significant limitations that point out the need for additional modalities. Herbal/natural products, particularly those based on green tea extract, offer promise as adjuncts or alternatives to current interventions and warrant further evaluation in well-controlled human trials.


title: [Epidemiological profile of pandemic influenza A cases in the south of Santa Catarina state, Brazil, in 2009].

text: OBJECTIVE To describe the epidemiological features of influenza A (H1N1) cases in 14 municipalities of Santa Catarina, Brazil, during the 2009 pandemic. METHOD This cross-sectional study focused on suspected cases of H1N1 reported to the 20th Santa Catarina State Health Administration during the 2009 pandemic between July to September. Data were collected by epidemiological surveillance officers from the Brazilian communicable diseases information system SINAN. H1N1 cases were confirmed by laboratory testing (positive RT-PCR) or signs and symptoms characteristic of severe acute respiratory syndrome. RESULTS During the pandemic period, 1 149 suspected cases of influenza A were notified, of which 560 (48.6%) were confirmed. That translated into an incidence of 241.9/100 000 population. Mean age for confirmed cases was 29.5 ± 17.1 years, vs. 32.2 ± 20 years for those ruled out (P = 0.03). Of the total confirmed cases, 37.1% were hospitalized, with a hospital incidence rate of 89.9/100 000 people and lethality rate of 5.6/100 000 population. Age < 30 years, symptoms of fever, cough and dyspnea, and death were independently associated with influenza A infection (P < 0.05). There were no associations between confirmed cases and any comorbidities. CONCLUSION The studied sample differed from the national profile of influenza A cases in Brazil by the absence of associated comorbidities. However, it was similar to the national profile in terms of the young age of cases and the significant association with fever, cough, and dyspnea. An extension of the annual immunization campaign (currently focused on risk groups) to the overall population should be considered.


title: Bacteria as potential tools in bioterrorism, with an emphasis on bacterial toxins.

text: The threat of bioterrorism remains a reality worldwide and, although of low probability, an attack would be a high-consequence event. Microbes are available to individuals with appropriate contacts and even many low-grade bacterial pathogens can severely affect health. Toxins provide bacteria with a system of defence that is often detrimental to humans and their versatility makes them potential tools of bioterrorism. It should be remembered that the aim of terrorism is not always to kill but rather to strike fear into peoples lives. Therefore, agents such as botulinum and cholera toxin could be used, which may not cause significant mortality but would cause widespread panic and potentially high morbidity. Importantly, no state can ever be fully prepared for a response and it is probable that no state ever could be. It is for this reason that biological agents are so attractive as weapons.


title: Impact of a Transfusion-free Program on Patients Undergoing Pancreaticoduodenectomy.

text: Patients undergoing pancreaticoduodenectomy (PD) often require transfusion. However, transfusion-related complications and decreased blood donation in Korea encourage the development of new treatment strategies for PD patients. Although transfusion-free (TF) operation is thought to be beneficial, results supporting its beneficial effects are lacking. The aim of our study was to demonstrate the impact on PD patients of a TF program. From December 2003 to April 2013, 80 consecutive patients with periampullary lesions underwent PD performed. These patients were divided into two groups as follows: 39 PD patients in the "before TF program" (Group 1) and 41 PD patients in the "after TF program" (Group 2). Among patients in Group 2, patients who agreed with the TF program were enrolled and proceed with the TF program prospectively. Participants in the TF program had perioperative blood augmentation and intraoperative acute normovolemic hemodilution. The perioperative data were compared with the two groups. The mean preoperative hemoglobin, operative times, and operative blood loss showed no significance between two groups. The mean postoperative hemoglobin was lower in Group 2 (11.7 g/dL vs 10.9 g/dL, P = 0.038). The mean amount of blood transfusion was significantly lower in Group 2. (950.8 mL vs 124.9 mL, P = 0.009). The TF program considerably decreases the amount of perioperative blood transfusion. The overall perioperative course and complication rate in the TF group were not inferior to those in the non-TF group. The TF program appears safe and should be considered in PD patients.


title: Persistence of viral RNA in the central nervous system of mice inoculated with MHV-4.

text: In order to study the role that viral persistence may play in chronic central nervous system (CNS) disease induced by murine coronaviruses, we have used the reverse transcriptase-polymerase chain reaction (RT-PCR) to study viral RNA in the brains of mice after intracerebral inoculation of JHM virus (JHMV or MHV-4). Quantitative RT-PCR showed that JHMV RNA decreased from approximately 2 ng/ug total brain RNA at day 6 post-inoculation (PI) to 0.1 pg/ug total brain RNA at 360 days PI. Double-stranded viral RNA could be detected up to day 20 PI. By the selective use of upstream or downstream primers during the RT step, it was possible to measure negative sense and positive sense JHMV RNA respectively, and we found that there was a marked rise in the ratio of positive to negative sense JHMV RNA after day 13 PI. Analysis of amplified products by dideoxy DNA sequencing showed that the characteristic mutation of our input virus (at position 3340 of gene 3) is maintained to at least day 42 PI. Taken together, these results favor a model of JHMV persistence in vivo in which viral RNA is present as double stranded forms initially and predominantly as single stranded, positive sense forms at late timepoints. Further analysis of this model in quantitative terms may contribute to our understanding of the biological significance of coronavirus persistence in the CNS.


title: Evolution of an emergency department screening questionnaire for severe acute respiratory syndrome.

text: OBJECTIVES To describe the screening tool that was used to screen for severe acute respiratory syndrome (SARS), the three revisions that were made, and the factors that led to these revisions. On March 13, 2003, on receiving notification of an outbreak of atypical pneumonia, nurses from the study emergency department (ED) started screening patients for the disease that became known as SARS. METHODS The ED nurses started with a simple screening tool that was incorporated into triage. The screening tool was later revised into a questionnaire. An outdoor screening station was set up and patients were subsequently screened before triage. After the patients were screened, they were assigned to different risk areas, where triage and treatment were rendered. Two further revisions were made to the questionnaire. RESULTS From March 13 to May 31, 11457 patients were screened. Version One of the screening questionnaire was used to screen 72 patients from March 13 to 17, Version Two screened 93 patients from March 18 to 21, Version Three screened 2909 patients from March 22 to April 8, and Version Four screened 8383 patients from April 9 to May 31. There was a significant (p < 0.05) downward trend in the proportion of admissions. Among those discharged from the ED, 0.28% reattended and were later confirmed to have SARS. CONCLUSIONS The screening tool underwent three major revisions in response to new information. By keeping it relatively simple, user-friendly, and regularly updated, nurses were able to screen patients rapidly. Risk categorization ensured that no cross-infection occurred among patients and that no one contracted SARS in the ED.


title: Collaborative colorectal cancer screening: a successful quality improvement initiative.

text: PROBLEM Low screening and referral rates for colorectal cancer at a primary care clinic suggest the need for alternative methods to identify patients and complete the screening process. DESIGN A review of >5000 medical charts established baseline screening and referral data. After a 3-month trial of a screening protocol, the research team conducted a follow-up medical chart review to determine referral levels. BACKGROUND AND SETTING The clinic is an 8-physician primary care facility in Southlake, Texas, and is one of 36 clinics affiliated with HealthTexas Provider Network. KEY MEASURES FOR IMPROVEMENT The goal was to increase referrals for colorectal cancer to at least 85% among patients aged 50 to 75 years. STRATEGIES FOR IMPROVEMENT The entire staff of the primary care clinic and the gastroenterology office became involved in the referral process. The team used simple tools such as chart stickers to draw attention to patients requiring screening, generation of referral forms that were numbered for follow-up and faxed to the gastroenterologists, and patient educational material on colorectal cancer screening. These tools were designed to overcome specific barriers to successful screening that the team had identified. EFFECTS OF CHANGE Referrals for sigmoidoscopy, colonoscopy, and double-contrast barium enema increased from 47% to 86%. Fecal occult blood testing was arranged for additional patients through the primary care office. Revenues related to colonoscopies increased by about 50% for the gastroenterologist group, the hospital, and the pathology group affiliated with Southlake Family Medicine. LESSONS LEARNED This colorectal cancer screening protocol succeeded in its 3-month trial because it was collaborative, opportunistic, simple, and made good business sense. The protocol is now being implemented at other HealthTexas Provider Network offices.


title: Nucleic-acid-based antiviral agents against positive single-stranded RNA viruses.

text: Positive single-stranded RNA viruses constitute a broad and prevalent group of pathogens that threaten human health and life worldwide. While effective vaccines have been developed for some, such as poliovirus and hepatitis A, others such as coxsackievirus, severe acute respiratory syndrome coronavirus (SARS-CoV) and West Nile virus have no accredited drug treatments. Antisense technologies, which encompass small interfering RNA, antisense oligonucleotides, ribozymes and their chemically modified analogs, involve small sequence-specific nucleic-acid-based molecules that inhibit viral replication at the level of translation. Many antisense oligomers are proven antiviral agents in vitro. In this review, iwe provide an overview of the antiviral antisense field, highlighting specific studies of interest over the past several years, using our experience with coxsackievirus B3 as a reference point. Overall, both the challenges and successes of existing antisense therapies for positive single-stranded RNA viruses can be paralleled to those for other virus groups, and vice versa.


title: Reverse transcriptase polymerase chain reaction-based diagnosis and molecular characterization of a new rat coronavirus strain.

text: BACKGROUND AND PURPOSE Rat coronaviruses (RCV) are highly infectious and spread rapidly through laboratory rat colonies, causing sneezing, nasal and ocular discharges, photophobia, and cervical swelling. Current diagnostic methods include serologic testing and histologic examination. During a recent rat coronavirus outbreak, we tested a rapid, noninvasive method of RCV diagnosis that involved use of reverse transcriptase-polymerase chain reaction (RT-PCR) analysis to detect RCV RNA on cages housing infected rats. METHODS The RT-PCR was used to detect RCV RNA in tissues from infected rats and on cages housing infected rats and to amplify portions of the RCV N, M, and S genes for molecular characterization. RESULTS The RT-PCR detected RCV RNA on cages and in tissues from infected rats. The RCV-NJ N gene is most closely related to the MHV-Y N gene. The M proteins of RCV-NJ and RCV-SDA are 99% homologous, and the six RCV S protein fragments are 97 to 100% homologous. CONCLUSIONS Use of RT-PCR with cage-swab specimens was capable of diagnosing RCV infection in and viral excretion from rats. Additionally, molecular characterization of the N, M, and S genes of RCV-NJ provided baseline information that can be used in performing further epidemiologic studies.


title: Prevalence and genetic diversity of coronaviruses in bats from China.

text: Coronaviruses can infect a variety of animals including poultry, livestock, and humans and are currently classified into three groups. The interspecies transmissions of coronaviruses between different hosts form a complex ecosystem of which little is known. The outbreak of severe acute respiratory syndrome (SARS) and the recent identification of new coronaviruses have highlighted the necessity for further investigation of coronavirus ecology, in particular the role of bats and other wild animals. In this study, we sampled bat populations in 15 provinces of China and reveal that approximately 6.5% of the bats, from diverse species distributed throughout the region, harbor coronaviruses. Full genomes of four coronavirues from bats were sequenced and analyzed. Phylogenetic analyses of the spike, envelope, membrane, and nucleoprotein structural proteins and the two conserved replicase domains, putative RNA-dependent RNA polymerase and RNA helicase, revealed that bat coronaviruses cluster in three different groups: group 1, another group that includes all SARS and SARS-like coronaviruses (putative group 4), and an independent bat coronavirus group (putative group 5). Further genetic analyses showed that different species of bats maintain coronaviruses from different groups and that a single bat species from different geographic locations supports similar coronaviruses. Thus, the findings of this study suggest that bats may play an integral role in the ecology and evolution of coronaviruses.


title: Essential veterinary education in zoological and wildlife medicine: a global perspective.

text: The current veterinary curriculum leaves graduates ill-equipped for careers in the field of zoological and wildlife medicine. Further postgraduate training is required to be an effective zoo or wildlife veterinarian. However, whether or not students choose to specialise in this field at a later date, the veterinary curriculum should cover several issues that are related to wildlife and zoo animals, including conservation biology, zoology, behaviour, physiology and conservation medicine. These subjects are essential, as we are preparing students to work in a world in which there is a global trade in wild animals, an increasing number of emerging infectious diseases and numerous environmental threats (habitat fragmentation, climate change) linked to anthropogenic change. Veterinary students should also be exposed to new opportunities to identify field and laboratory tools for the management and possible treatment of diseases in captive and wild populations and ecosystems using both in situ and ex situ approaches to conservation.


title: The impact of the Hand Hygiene New Zealand programme on hand hygiene practices in New Zealand's public hospitals.

text: AIM To detail the progress made by Hand Hygiene New Zealand (HHNZ) since 2011 and also describe the challenges experienced along the way and the factors required for delivery of a successful hand hygiene programme at a national level. METHOD HHNZ is a multimodal culture-change programme based on the WHO '5 moments for hand hygiene' approach. The key components of the programme include clinical leadership, auditing of hand hygiene compliance with thrice yearly reporting of improvement in hand hygiene practice, biannual reporting of the outcome marker, healthcare-associated Staphylococcus aureus bacteraemia (HA-SAB), effective communication with key stakeholders and the use of the front-line ownership (FLO) principles for quality improvement. RESULTS The nationally aggregated hand hygiene compliance has increased from 62% in June 2012 to 81% in March 2016. There has been improvement across all 'moments', all healthcare worker groups and a range of different clinical specialties. The rate of HA-SAB has remained stable. CONCLUSION The HHNZ programme has led to significant improvements in hand hygiene practice in DHBs throughout New Zealand. The principles of FLO are now widely used to drive hand hygiene improvement in New Zealand DHBs.


Temporal regulation of RNA synthesis of bovine coronavirus.


title: Computed tomography screening for lung cancer: where are we now?

text: Low-dose computed tomography (LDCT) screening has been shown to result in detection of earlier-stage lung cancers, with a 20% reduction in cancer-related deaths. LDCT screening offers significant potential benefits to selected patients; however, many questions remain, including questions about the applicability of lung cancer screening in clinical practice.


title: Genetic, geographical and temporal variation of porcine reproductive and respiratory syndrome virus in Illinois.

text: Porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 gene sequences were generated by RT-PCR from 55 field isolates collected in Illinois and eastern Iowa. Spatial and temporal patterns of genetic variation in the virus were examined on a local geographical scale in order to test the hypothesis that the genetic similarity of PRRSV isolates (measured as their percentage pairwise ORF5 nucleotide similarity) was positively correlated with their geographical proximity. Levels of genetic variability in the Illinois/eastern Iowa PRRSV sample were similar to levels of variability seen across broader geographical regions within North America. The genetic similarity of isolates did not correlate with their geographical distance. These results imply that the movement of PRRSV onto farms does not generally occur via distance-limited processes such as wind or wildlife vectors, but more typically occurs via the long-distance transport of animals or semen. Genetic distances between PRRSV isolates collected from the same farms at different times increased as the time separating the collection events increased. This result implies rapid movement of new genetic types of PRRSV into and out of farms. PRRSV ORF5 displayed a pattern of third-codon-position diversity bias that was not evident in a geographically comparable sample of pseudorabies virus (a swine alphaherpesvirus) gC gene sequences. This result provides evidence that PRRSV ORF5 is experiencing stabilizing selection against structural novelty. Despite high genetic variability at all geographical levels, PRRSV ORF5 nevertheless contained potentially antigenic regions that were invariant at the amino acid level. These regions should make effective vaccine targets if they prove to be immunogenic.


title: Incidence of upper respiratory tract Mycoplasma pneumoniae infections among outpatients in Rhône-Alpes, France, during five successive winter periods.

text: In this prospective study, nasal swab samples from patients with acute respiratory infections were evaluated for the presence of Mycoplasma pneumoniae. This PCR-plus-hybridization-based detection was associated with the detection of other viral agents. During the five winter surveillance periods, 3,897 samples were collected by 75 medical practitioners participating in the Groupe Régional d'Observation de la Grippe surveillance network in Rhône-Alpes (France). M. pneumoniae was detected in 283 samples (7.3%); its rate of detection ranged from 10.1 to 2.0% over the five periods, and it was the second most frequently isolated pathogen during the survey, after influenza A. Three high-prevalence winters were observed, yielding an early winter peak of M. pneumoniae infection which was observed in all age groups. No statistically significant difference was detected between rates of infections in the different age groups, but M. pneumoniae infection was significantly related to lower respiratory tract infection during periods of high prevalence. This study defined the frequency of M. pneumoniae detection from nasal swab specimens in patients with acute respiratory infections, confirming its high prevalence and the presence of large outbreaks due to this pathogen.


title: Efficacy of isolated colostral IgA, IgG, and IgM(A) to protect neonatal pigs against the coronavirus of transmissible gastroenteritis.

text: To ascertain what class of immunoglobulin (Ig; IgA, IgG, or IgM) is most efficacious in protection, a large quantity of colostrum from sows immunized with virulent transmissible gastroenteritis (TGE) virus was fractionated by chromatographic and gel filtration methods. The isolated IgG, IgA, and IgM(A) had specific virus-neutralizing activities of 1:7.6, 1:342, and 1:302 per milligram of protein, respectively. Each Ig was fed to groups of hysterectomy-derived colostrum-deprived neonatal pigs before and after exposure (challenge) with virulent TGE virus. The 7 pigs fed IgG survived the challenge exposure, but 2 of 7 fed IgA and 1 of 7 fed IgM(A) died of TGE. Three of the survivor pigs that had been fed IgG and 2 of the survivor pigs that had been fed IgA had increased serum antibody titers between 8 and 19 days after challenge exposure, but none of the survivor pigs fed IgM(A) had TGE antibody. In contrast, 12 of 14 virus-control pigs died of TGE and the 2 survivors had antibody conversion. The data show that all 3 Ig classes in immune colostrum will protect neonatal pigs against exposure with virulent TGE virus.


title: Specific binding of host cellular proteins to multiple sites within the 3' end of mouse hepatitis virus genomic RNA.

text: The initial step in mouse hepatitis virus (MHV) RNA replication is the synthesis of negative-strand RNA from a positive-strand genomic RNA template. Our approach to begin studying MHV RNA replication is to identify the cis-acting signals for RNA synthesis and the proteins which recognize these signals at the 3' end of genomic RNA of MHV. To determine whether host cellular and/or viral proteins interact with the 3' end of the coronavirus genome, an RNase T1 protection/gel mobility shift electrophoresis assay was used to examine cytoplasmic extracts from mock- and MHV-JHM-infected 17Cl-1 murine cells for the ability to form complexes with defined regions of the genomic RNA. We demonstrated the specific binding of host cell proteins to multiple sites within the 3' end of MHV-JHM genomic RNA. By using a set of RNA probes with deletions at either the 5' or 3' end or both ends, two distinct binding sites were located. The first protein-binding element was mapped in the 3'-most 42 nucleotides of the genomic RNA [3' (+42) RNA], and the second element was mapped within an 86-nucleotide sequence encompassing nucleotides 171 to 85 from the 3' end of the genome (171-85 RNA). A single potential stem-loop structure is predicted for the 3' (+)42 RNA, and two stem-loop structures are predicted for the 171-85 RNA. Proteins interacting with these two elements were identified by UV-induced covalent cross-linking to labeled RNAs followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The RNA-protein complex formed with the 3'-most 42 nucleotides contains approximately five host polypeptides, a highly labeled protein of 120 kDa and four minor species with sizes of 103, 81, 70, and 55 kDa. The second protein-binding element, contained within a probe representing nucleotides 487 to 85 from the 3' end of the genome, also appears to bind five host polypeptides, 142, 120, 100, 55, and 33 kDa in size, with the 120-kDa protein being the most abundant. The RNA-protein complexes observed with MHV-infected cells in both RNase protection/gel mobility shift and UV cross-linking assays were identical to those observed with uninfected cells. The possible involvement of the interaction of host proteins with the viral genome during MHV replication is discussed.


title: Genome organization and reverse genetic analysis of a type I feline coronavirus.

text: In this study we report the complete sequence and genome organization of the serotype I feline coronavirus (FCoV) strain Black. Furthermore, a reverse genetic system was established for this FCoV strain by cloning a full-length cDNA copy into vaccinia virus. This clone served as basis for the generation of recombinant FCoV (recFCoV) that was shown to bear the same features in vitro as the parental FCoV. Using this system, accessory 3abc genes in the FCoV genome were replaced by green fluorescent protein (recFCoV-GFP) and Renilla luciferase genes (recFCoV-RL). In addition, we showed that feline CD14(+) blood monocytes and dendritic cells can be easily detected after infection with recFCoV-GFP. Thus, our established reverse genetic system provides a suitable tool to study the molecular biology of serotype I FCoV.


title: American College of Rheumatology 2012 recommendations for the use of nonpharmacologic and pharmacologic therapies in osteoarthritis of the hand, hip, and knee.

text: OBJECTIVE To update the American College of Rheumatology (ACR) 2000 recommendations for hip and knee osteoarthritis (OA) and develop new recommendations for hand OA. METHODS A list of pharmacologic and nonpharmacologic modalities commonly used to manage knee, hip, and hand OA as well as clinical scenarios representing patients with symptomatic hand, hip, and knee OA were generated. Systematic evidence-based literature reviews were conducted by a working group at the Institute of Population Health, University of Ottawa, and updated by ACR staff to include additions to bibliographic databases through December 31, 2010. The Grading of Recommendations Assessment, Development and Evaluation approach, a formal process to rate scientific evidence and to develop recommendations that are as evidence based as possible, was used by a Technical Expert Panel comprised of various stakeholders to formulate the recommendations for the use of nonpharmacologic and pharmacologic modalities for OA of the hand, hip, and knee. RESULTS Both “strong” and “conditional” recommendations were made for OA management. Modalities conditionally recommended for the management of hand OA include instruction in joint protection techniques, provision of assistive devices, use of thermal modalities and trapeziometacarpal joint splints, and use of oral and topical nonsteroidal antiinflammatory drugs (NSAIDs), tramadol, and topical capsaicin. Nonpharmacologic modalities strongly recommended for the management of knee OA were aerobic, aquatic, and/or resistance exercises as well as weight loss for overweight patients. Nonpharmacologic modalities conditionally recommended for knee OA included medial wedge insoles for valgus knee OA, subtalar strapped lateral insoles for varus knee OA, medially directed patellar taping, manual therapy, walking aids, thermal agents, tai chi, self management programs, and psychosocial interventions. Pharmacologic modalities conditionally recommended for the initial management of patients with knee OA included acetaminophen, oral and topical NSAIDs, tramadol, and intraarticular corticosteroid injections; intraarticular hyaluronate injections, duloxetine, and opioids were conditionally recommended in patients who had an inadequate response to initial therapy. Opioid analgesics were strongly recommended in patients who were either not willing to undergo or had contraindications for total joint arthroplasty after having failed medical therapy. Recommendations for hip OA were similar to those for the management of knee OA. CONCLUSION These recommendations are based on the consensus judgment of clinical experts from a wide range of disciplines, informed by available evidence, balancing the benefits and harms of both nonpharmacologic and pharmacologic modalities, and incorporating their preferences and values. It is hoped that these recommendations will be utilized by health care providers involved in the management of patients with OA.


title: The avian coronavirus infectious bronchitis virus undergoes direct low-pH-dependent fusion activation during entry into host cells.

text: Coronaviruses are the causative agents of respiratory disease in humans and animals, including severe acute respiratory syndrome. Fusion of coronaviruses is generally thought to occur at neutral pH, although there is also evidence for a role of acidic endosomes during entry of a variety of coronaviruses. Therefore, the molecular basis of coronavirus fusion during entry into host cells remains incompletely defined. Here, we examined coronavirus-cell fusion and entry employing the avian coronavirus infectious bronchitis virus (IBV). Virus entry into cells was inhibited by acidotropic bases and by other inhibitors of pH-dependent endocytosis. We carried out fluorescence-dequenching fusion assays of R18-labeled virions and show that for IBV, coronavirus-cell fusion occurs in a low-pH-dependent manner, with a half-maximal rate of fusion occurring at pH 5.5. Fusion was reduced, but still occurred, at lower temperatures (20 degrees C). We observed no effect of inhibitors of endosomal proteases on the fusion event. These data are the first direct measure of virus-cell fusion for any coronavirus and demonstrate that the coronavirus IBV employs a direct, low-pH-dependent virus-cell fusion activation reaction. We further show that IBV was not inactivated, and fusion was unaffected, by prior exposure to pH 5.0 buffer. Virions also showed evidence of reversible conformational changes in their surface proteins, indicating that aspects of the fusion reaction may be reversible in nature.


title: Assessing the quality of a web-based learning system for nurses.

text: Continuing professional education is essential for nurses to update their clinical skills and knowledge to meet the complex demands of current patient care. Compared to traditional in-class continuing education, a web-based learning system is efficient for nurses with a three shift-working schedule and is timely to deliver knowledge about newly emerging epidemics such as Severe Acute Respiratory Syndrome. Many studies reveal the advantages of various web-based learning systems but seldom evaluate them from the perspective of quality. This study develops an instrument to assess the quality of a web-based learning system for nurses' continuing education based on the quality dimensions of a mature information systems success model. The research results show that all indicators of the instrument provide a fit to the quality measurement of a web-based learning system and have high reliability and validity. Based on the research findings, implications and limitations are discussed.


title: Zinc-binding of the cysteine-rich domain encoded in the open reading frame of 1B of the RNA polymerase gene of coronavirus.

text: We cloned and sequenced the second open reading frame of the RNA polymerase gene, ORF1b, of bovine coronavirus. In the region representing nucleotide positions 4919-5677 upstream from the initiation codon of the 32K non-structural protein gene, we identified two putative functional domains. One of these domains contained four leucine residues repeated exactly in every seventh position, and the other domain represented a cluster of cysteine and histidine residues. The DNA sequence representing these domains was cloned and expressed in Escherichia coli as fusion proteins with glutathione S-transferase from Schistosoma japonicum. A high level expression of the cysteine-rich domain was achieved as a fusion protein when the bacterial culture was induced with IPTG. In a solid phase zinc binding assay using the recombinant fusion protein, we found that the protein containing the cysteine-rich domain was able to bind to radioactive zinc in vitro, demonstrating that the polypeptide encoded by the ORF1b of coronavirus is a zinc-binding protein.