title: Ensuring a broad and inclusive approach: a provincial perspective on pandemic preparedness.

text: The SARS crisis revealed critical gaps in Ontario's health emergency response capacity, and identified, in the starkest terms possible, the need for improved emergency response planning. This article reviews the development of the Ontario Health Plan for an Influenza Pandemic (OHPIP), released in June 2005. Some key points arising from the provincial planning process include the necessity to: ensure a broad and inclusive development process; ensure the pandemic plan identifies: 1) clear roles and responsibilities of federal, provincial/territorial and municipal levels of government, 2) the approach to occupational health and safety issues and ethical decision-making, 3) a communications strategy linking all affected sectors and levels of government and health sector; 4) any commitments to antiviral stockpiling, vaccine and antiviral allocation and use, and an approach for drug delivery from provincial stockpiles to local public health units; 5) health human resource management and supplementation; and 6) key programs/services to be scaled back to maximize surge capacity; address best practices (e.g., involve all sectors of the health care system at the outset, acquire strategic expertise, coordinate/advocate with broader emergency response system, etc); and, outline future stages that include strengthening the delivery of clinical care to influenza cases; clarifying the role of primary care practitioners during a pandemic; leveraging Ontario's significant e-Health investments.


title: No evidence for quasispecies populations during persistence of the coronavirus mouse hepatitis virus JHM: sequence conservation within the surface glycoprotein gene S in Lewis rats.

text: The surface glycoprotein S (spike) of coronaviruses is believed to be an important determinant of virulence and displays extensive genetic polymorphism in cell culture isolates. This led us to consider whether the observed heterogeneity is reflected by a quasispecies distribution of mutated RNA molecules within the infected organ. Coronavirus infection of rodents is a useful model system for investigating the pathogenesis of virus-induced central nervous system (CNS) disease. Here, we investigated whether genetic changes in the S gene occurred during virus persistence in vivo. We analysed the variability of S gene sequences directly from the brain tissue of Lewis rats infected with the coronavirus mouse hepatitis virus (MHV) variant JHM-Pi using RT-PCR amplification methods. The S gene sequence displayed a remarkable genetic stability in vivo. No evidence for a quasispecies distribution was found by sequence analysis of amplified S gene fragments derived from the CNS of Lewis rats. Furthermore, the S gene also remained conserved under the selection pressure of a neutralizing antibody. Only a few mutations predicted to result in amino acid changes were detected in single clones. The changes were not represented in the consensus sequence. These results indicate that to retain functional proteins under the constraints of a persistent infection in vivo, conservation of sequence can be more important than heterogeneity.


Response of the clinical microbiology laboratory to emerging (new) and reemerging infectious diseases.


title: Prevalence of feline leukaemia virus and antibodies to feline immunodeficiency virus and feline coronavirus in stray cats sent to an RSPCA hospital.

text: A total of 517 stray cats at an RSPCA veterinary hospital were tested for feline leukaemia virus (FeLV), feline coronavirus (FCoV) and feline immunodeficiency virus (FIV). The prevalence of FeLV was 3.5 per cent in all the cats, 1.4 per cent in healthy cats and 6.9 per cent in sick cats. FeLV positivity was associated only with disease of non-traumatic origin. Antibodies to FCoV were present in 22.4 per cent of the cats, and their prevalence was significantly higher in cats over two years old and in feral/semiferal cats. The prevalence of antibodies to FIV was 10.4 per cent in all the cats, 4.9 per cent in healthy cats and 16.7 per cent in sick cats. The prevalence of FIV antibodies was significantly higher in entire males and neutered males than in females, in cats over two years old compared with younger cats, and in cats suffering disease of non-traumatic origin rather than in healthy cats or cats suffering only from trauma. Sex, age and health status were each independently highly associated with FIV antibodies.


title: Elderly suicide and the 2003 SARS epidemic in Hong Kong.

text: BACKGROUND Hong Kong was struck by the community outbreak of Severe Acute Respiratory Syndrome (SARS) in 2003. In the same year, the elderly suicide rate in Hong Kong showed a sharp upturn from a previous downward trend. METHODS Secondary analyses using Poisson Regression Models on the suicide statistics from the Census and Statistics Department of the Hong Kong Government were performed. RESULTS In a Poisson Regression Model on the annual suicide rates in elders aged 65 and over in years 1986-2003, 2002 served as the reference year. Suicide rates in 1986-1997 were significantly higher than the reference year, with an Incident Rate Ratio (IRR) of 1.34 to 1.61. However, rates in 1998-2001 did not differ from the reference year significantly, representing stabilization of suicide rates for 4 years after 1997. The elderly suicide rate increased to 37.46/100,000 in 2003, with an IRR of 1.32 (p=0.0019) relative to 2002. Such trend is preserved when female elderly suicide rates in 1993-2003 were analyzed, while suicide rates in elderly men and younger age groups did not follow this pattern. DISCUSSIONS Mechanistic factors such as breakdown of social network and limited access to health care might account for the findings. These factors could have potentiated biopsychosocial risk factors for suicide at individual levels, particularly in elderly. Female elders, by way of their previous readiness to utilize social and health services instituted in the past decade, are thus more susceptible to the effects of temporary suspension of these services during the SARS epidemic. CONCLUSIONS The SARS epidemic was associated with increased risk of completed suicide in female elders, but not in male elders or the population under 65 years of age.


title: Characterization of coronavirus DI RNA packaging.

text: Studies of defective interfering (DI) RNAs of mouse hepatitis virus (MHV), suggest that a 69 nt-long packaging signal, which is located about 20 kb from the 5'-end of the 31 kb-long MHV genomic RNA, is necessary and sufficient for MHV genomic RNA packaging into MHV particles. We demonstrated that use of a low pH culture medium combined with subsequent ultrafiltration increased MHV infectivity about 60 times over MHV preparations grown in neutral medium. Using this virus concentration procedure, we successfully prepared DI particle-rich MHV preparations. Characterization of virus samples released from the cells infected with DI particle-rich MHV revealed that infectious MHV genomic RNA was not required for packaging of DI RNAs. These data suggested that interaction of the DI packaging signal with an unidentified region(s) of helper virus genomic RNA is unlikely, and therefore unlikely to facilitate the packaging of MHV DI RNA into the MHV virion. Rather, both DI RNA and MHV genomic RNA probably use the packaging signal for RNA packaging.


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title: The Danish Ventral Hernia Database - a valuable tool for quality assessment and research.

text: AIM The Danish Ventral Hernia Database (DVHD) provides national surveillance of current surgical practice and clinical postoperative outcomes. The intention is to reduce postoperative morbidity and hernia recurrence, evaluate new treatment strategies, and facilitate nationwide implementation of evidence-based treatment strategies. This paper describes the design and purpose of DVHD. STUDY POPULATION Adult (≥18 years) patients with a Danish Civil Registration Number and undergoing surgery under elective or emergency conditions for ventral hernia in a Danish surgical department from 2007 and beyond. A total of 80% of all ventral hernia repairs performed in Denmark were reported to the DVHD. MAIN VARIABLES Demographic data (age, sex, and center), detailed hernia description (eg, type, size, surgical priority), and technical aspects (open/laparoscopic and mesh related factors) related to the surgical repair are recorded. Data registration is mandatory. Data may be merged with other Danish health registries and information from patient questionnaires or clinical examinations. DESCRIPTIVE DATA More than 37,000 operations have been registered. Data have demonstrated high agreement with patient files. The data allow technical proposals for surgical improvement with special emphasis on reduced incidences of postoperative complications, hernia recurrence, and chronic pain. CONCLUSION DVHD is a prospective and mandatory registration system for Danish surgeons. It has collected a high number of operations and is an excellent tool for observing changes over time, including adjustment of several confounders. This national database registry has impacted on clinical practice in Denmark and led to a high number of scientific publications in recent years.


title: X-ray structures of the N- and C-terminal domains of a coronavirus nucleocapsid protein: implications for nucleocapsid formation.

text: Coronaviruses cause a variety of respiratory and enteric diseases in animals and humans including severe acute respiratory syndrome. In these enveloped viruses, the filamentous nucleocapsid is formed by the association of nucleocapsid (N) protein with single-stranded viral RNA. The N protein is a highly immunogenic phosphoprotein also implicated in viral genome replication and in modulating cell signaling pathways. We describe the structure of the two proteolytically resistant domains of the N protein from infectious bronchitis virus (IBV), a prototype coronavirus. These domains are located at its N- and C-terminal ends (NTD and CTD, respectively). The NTD of the IBV Gray strain at 1.3-A resolution exhibits a U-shaped structure, with two arms rich in basic residues, providing a module for specific interaction with RNA. The CTD forms a tightly intertwined dimer with an intermolecular four-stranded central beta-sheet platform flanked by alpha helices, indicating that the basic building block for coronavirus nucleocapsid formation is a dimeric assembly of N protein. The variety of quaternary arrangements of the NTD and CTD revealed by the analysis of the different crystal forms delineates possible interfaces that could be used for the formation of a flexible filamentous ribonucleocapsid. The striking similarity between the dimeric structure of CTD and the nucleocapsid-forming domain of a distantly related arterivirus indicates a conserved mechanism of nucleocapsid formation for these two viral families.


title: Genetic analysis of feline caliciviruses associated with a hemorrhagic-like disease.

text: Feline calicivirus (FCV) is 1 of the most common causes of upper respiratory tract disease in cats. Other disease syndromes associated with FCV infection have been reported. Recently, calicivirus infection associated with a hemorrhagic-like disease leading to significant mortality in cats has been reported. The clinical signs are similar to those observed with the calicivirus of rabbit hemorrhagic disease. This study characterized 2 FCV isolates associated with hemorrhagic-like disease. Nucleotide sequencing of the complete genome has been done for these 2 isolates as well as for 4 additional isolates representing other disease syndromes. Previously reported sequence data for the entire genome of classical FCV (6 isolates) and a portion of the capsid gene for hemorrhagic-like FCV (3 isolates), isolated in different regions of United States were used in the genetic analysis. Sequence data were used to determine relationships among the isolates and any correlation with phenotype. Nucleotide sequence comparisons of the entire genome and individual open reading frames revealed high homology among all isolates. Data suggest that the virulence may have genetic determinants on the basis of phylogenetic clustering of the isolates associated with hemorrhagic-like disease.


title: Real-time quantitative broad-range PCR assay for detection of the 16S rRNA gene followed by sequencing for species identification.

text: Here we determined the analytical sensitivities of broad-range real-time PCR-based assays employing one of three different genomic DNA extraction protocols in combination with one of three different primer pairs targeting the 16S rRNA gene to detect a panel of 22 bacterial species. DNA extraction protocol III, using lysozyme, lysostaphin, and proteinase K, followed by PCR with the primer pair Bak11W/Bak2, giving amplicons of 796 bp in length, showed the best overall sensitivity, detecting DNA of 82% of the strains investigated at concentrations of < or =10(2) CFU in water per reaction. DNA extraction protocols I and II, using less enzyme treatment, combined with other primer pairs giving shorter amplicons of 466 bp and 342 or 346 bp, respectively, were slightly more sensitive for the detection of gram-negative but less sensitive for the detection of gram-positive bacteria. The obstacle of detecting background DNA in blood samples spiked with bacteria was circumvented by introducing a broad-range hybridization probe, and this preserved the minimal detection limits observed in samples devoid of blood. Finally, sequencing of the amplicons generated using the primer pair Bak11W/Bak2 allowed species identification of the detected bacterial DNA. Thus, broad-spectrum PCR targeting the 16S rRNA gene in the quantitative real-time format can achieve an analytical sensitivity of 1 to 10 CFU per reaction in water, avoid detection of background DNA with the introduction of a broad-range probe, and generate amplicons that allow species identification of the detected bacterial DNA by sequencing. These prerequisites are important for its application to blood-containing patient samples.


title: Isolation and trypsin-enhanced propagation of turkey enteric (bluecomb) coronaviruses in a continuous human rectal adenocarcinoma cell line.

text: Turkey enteric coronavirus (TCV) from intestinal contents of diarrheal poults was isolated and serially propagated in HRT-18 cells, an established cell line derived from a human rectal adenocarcinoma. In these cells, TCV induced cytopathic changes, including polykaryocytosis, which depended on trypsin in the medium and incubation at 41 C. Viral antigens could be demonstrated in the cytoplasm by immunofluorescence, and extracellular virus was detected by an ELISA and negative electron microscopy. The cell-free virus had characteristics of TCV: shape, surface projections, buoyant density of 1.18 to 1.20 g/ml in sucrose, and hemagglutination of rat RBC. The one-step growth curve was complete by postinoculation hours 14 to 16, and maximal titers reached 9 to 9.5 log10 TCID50/ml during 5 passages, after which the titer remained stable. Electron microscopic examination of infected cell monolayers revealed budding of typical coronavirus particles through intracytoplasmic membranes and accumulation of complete virus in cytoplasmic vesicles. Late in the infection, aggregated progeny vial particles were detected near the outer surface of infected cells. One-day-old turkey poults inoculated orally with tissue culture-adapted TCV isolates developed mild to severe diarrhea.


title: [Calf coronavirus neonatal diarrhea. A literature review (author's transl)].

text: Calf coronavirus neonatal diarrhea. A literature reviewThe importance of the calf coronavirus in the etiology of neonatal diarrhea of calves has been reported many times from various countries. A literature review concerning this virus is presented in this paper. A detailed description of the pathogenesis, clinical signs and lesions of the disease, as well as the morphological, physicochemical, biological and antigenic characteristics of the virus are presented. The immunity of the calf against this virus and the principal diagnosis technics are also discussed.


title: ANN-based mapping of febrile subjects in mass thermogram screening: facts and myths.

text: Severe Acute Respiratory Syndrome (SARS) is a highly infectious disease caused by a coronavirus. Screening to detect potential SARS-infected subjects with elevated body temperature plays an important role in preventing the spread of SARS. Thermography is being used with ANN/AI to analyse the data collected from the designated SARS hospital in Singapore, and conclusive results are drawn. The current work evaluates the correlations (and classifications) between facial skin temperatures, including eye range and forehead, to aural temperature using a neural network (NN) approach, namely training backpropagation (BP) and Kohonen self-organizing map (SOM), to confirm the suitability of thermal imagers for human temperature screening. Both BP and SOM can form an opinion about the type of network that is better able to complement thermogram technology in fever diagnosis. This can produce better parameters for reducing the size of the NN classifier, while maintaining good classification accuracy. We observe that BP performs better than SOM NN. Confusion matrix (CM), an alternative display instrument, is able to process a high volume of input data and show the clustered output rapidly and accurately. The current research application will remain an interesting and useful reference for both local and overseas manufacturers of thermal scanners, users and various government and private establishments. As the elevation of body temperature is a common presenting symptom for many illnesses, including infectious diseases such as SARS, thermal imagers are useful and essential tools for mass screening of body temperature. This is true not only for SARS but also during other public health crises where widespread transmission of infection such as the danger of avian flu pandemic is a concern, in particular at places like hospitals and cross-border checkpoints.


title: Effects of angiotensin II receptor blockers on renal handling of uric acid in rats.

text: Elevated serum uric acid level has been associated with increased cardiovascular risk in hypertensive patients. Several angiotensin II receptor blockers exhibit differential effects on regulation of serum uric acid level in humans. We have demonstrated that some angiotensin II receptor blockers trans-stimulate the uptake of uric acid by human URAT1 and others inhibit the transport of uric acid mediated by human URAT1, OAT1, OAT3 and MRP4 in vitro. This study investigated the effects of candesartan, pratosartan and telmisartan on renal handling of uric acid in rats in vivo and in vitro. Candesartan (0.1 mg/kg) significantly decreased the urinary excretion of uric acid and increased the plasma uric acid concentration. The kidney candesartan level after low-dose treatment is close to that required to trans-stimulate uric acid uptake in vitro. Pratosartan exhibited dose-dependent hypouricemic and uricosuric effects, while telmisartan showed no effects on plasma uric acid level. Furthermore, we confirmed the effects of the tested drugs on uric acid transport by rat renal brush border membrane transporter(s) and basolateral Oat1 and Oat3. Effects of angiotensin II receptor blockers in rats may be mainly determined by their intrinsic effects (cis-inhibition and trans-stimulation) on uric acid reabsorption transporter(s) and their pharmacokinetic properties in rats.


title: A seroepidemiological study of the importance in cow-calf pairs of respiratory and enteric viruses in beef operations from northwestern Quebec.

text: Serum antibody analyses for bovine herpesvirus type 1 (BHV-1), bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), bovine coronavirus (BCV), and bovine rotavirus (BRV) were performed on 527 randomly selected cows, before calving, and on 407 three-week-old calves. In cows and calves, BCV and BRV were the most seroprevalent viruses (80% to 100% according to virus and vaccination status). Bovine respiratory syncytial virus was the least seroprevalent in the cows, independent of the vaccination status. In nonvaccinated cows the seroprevalence to BRSV was 36.7%, and 53.5% in cows vaccinated less than two weeks prior to collecting blood, and 67.6% in cows vaccinated two weeks or more prior to blood collection. In their calves, BHV-1 was the least seroprevalent, independent of the vaccination status. The serological status and antibody titers in calves were generally associated with those of the dam. The occurrence of respiratory diseases in the calves was associated with cow and calf serological profiles (BHV-1, BRSV and BCV in the nonvaccinated group, BHV-1, BVDV and BCV in the vaccinated group). The occurrence of diarrhea was not associated with cow and calf serological profiles but was negatively associated with high level calf serum IgG in the nonvaccinated group (odds ratio = 0.73). Bovine coronavirus and BRV were shed by 1.4% and 4.9% of calves in the nonvaccinated group, and by 0% and 9.9% of calves in the vaccinated group, respectively. Bovine rotavirus shedding was associated with fecal diarrheic consistency at the moment of fecal sampling but not with previous occurrence of diarrhea.


title: Transmission scenarios for Middle East Respiratory Syndrome Coronavirus (MERS-CoV) and how to tell them apart.

text: Detection of human cases of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) infection internationally is a global public health concern. Rigorous risk assessment is particularly challenging in a context where surveillance may be subject to under-ascertainment and a selection bias towards more severe cases. We would like to assess whether the virus is capable of causing widespread human epidemics, and whether self-sustaining transmission is already under way. Here we review possible transmission scenarios for MERS-CoV and their implications for risk assessment and control. We discuss how existing data, future investigations and analyses may help in reducing uncertainty and refining the public health risk assessment and present analytical approaches that allow robust assessment of epidemiological characteristics, even from partial and biased surveillance data. Finally, we urge that adequate data be collected on future cases to permit rigorous assessment of the transmission characteristics and severity of MERS-CoV, and the public health threat it may pose. Going beyond minimal case reporting, open international collaboration, under the guidance of the World Health Organization and the International Health Regulations, will impact on how this potential epidemic unfolds and prospects for control.


title: [Changes in pulmonary function in severe acute respiratory syndrome patients during convalescent period].

text: OBJECTIVE To investigate the changes and pattern of pulmonary function in severe acute respiratory syndrome (SARS) patients during convalescent period. METHODS Pulmonary function tests were performed in 26 SARS convalescent patients regularly every 3 months after their discharge from hospital. The significance of changes in pulmonary function indexes was analyzed. RESULTS Restrictive pulmonary ventilation function and diffusing dysfunction of the lung were found in one third of the patients during third to sixth month from onset. There was a little improvement in forced vital capacity (FVC), one second forced expiratory volume (FEV(1.0)), functional residual capacity (FRC) and residual volume (RV) as convalescent period was prolonged, but no difference was found between different stages (3-6 months, 6-9 months, 9-12 months, 12-15 months and 15-17 months). Meanwhile, the FEV(1.0)/FVC showed no significant change. However, there was an obvious improvement in total lung capacity (TLC) and diffusing capacity of the lung for carbon monoxide (DLCO) with the elapse of time, and also a significant difference was found between the later stage and the earlier stage. CONCLUSION Pulmonary dysfunction is found among some SARS patients after convalescence presenting mainly as restrictive ventilatory function and diffusing capacity abnormality. These dysfunctions would improve gradually with the elapse of time.


title: [Construction of recombinant Lactococcus lactis expressing porcine transmissible gastroenteritis spike glycoprotein and analysis of immunogenicity].

text: Transmissible gastroenteritis virus (TGEV), is an enteropathogenic coronavirus that causes a highly fatal acute diarrhea in newborn pigs. It's typically clinical manifestations consist of omitting, severe diarrhea, loss water and highly infectious disease. All kinds and ages of pigs can be infected. Particular, the mortality piglets under 3 weeks may reach 100% . The effective protection against TGEV requires the development of vaccines that are able to induce local mucosal immunization. Lactococcus lactis was selected as a bacterial carrier for the expression of TGEV spike glycoprotein. The gene of S glycoprotein was cloned into the Lactococcus lactis vectors pNZ8112. An approximately 2000 bps fragments of TGEV gene S that encompasses all the four major antigenic domains critical for neutralization was transformed into Lactococcus lactis NZ9000 by electroporation, resulting in the recombinant strain pNZ8112-Sa/NZ9000. The recombinant glycoprotein S was detected by SDS-PAGE and Western blot after induced by 1ng/mL nisin. The result indicated that the expressed product maintain the antigenicity of TGEV as expected. In order to detect the location of expressed protein, the yellow and green fluorescence of the recombinant strain pNZ8112-Sa/NZ9000 was detected by the IFA experiments, which indicated that the expressed recombinant protein was secreted and located on the surface of the bacterium cell. Oral immunization of BALB/c mice with recombinant strain that constitutively express the 66kDa fragment of the glycoprotein S, Specific anti-TGEV glycoprotein S secret immunoglobulin A (sIgA) antibodies were detected by indirect enzyme linked immunosorbent assay (ELISA) in the feces after immunization. It was showed that the mice immunized with pNZ8112-Sa/NZ9000 recombinant strain had produced clear antibody level anti TGEV, and which had provided important substance foundation and explored the feasibility of Lactobacillus as oral vaccine.


title: Conversion in laparoscopic cholecystectomy after gastric resection: a 15-year review.

text: BACKGROUND Gastrectomy or truncal vagotomy is known to increase the incidence of cholelithiasis. Many of these patients will become symptomatic, and the adhesions from their gastric resection may make laparoscopic cholecystectomy much more difficult. METHODS We prospectively assessed the data for the 15-year cumulative laparoscopic cholecystectomy experience of 1 surgeon at a university teaching hospital with respect to conversion and postoperative outcomes, with particular attention to patients having had previous gastric resections. RESULTS Patients with previous gastrectomies had similar operative times (mean 81.1, range 45-120 min), a higher conversion rate (64.2%) and a higher complication rate (35.7%) than those who had had other previous upper abdominal surgeries (mean 73.2, range 35-130 min, conversion 25% and complication 11.3%) and those without previous abdominal surgeries (mean 66.5, range 25-250 min, conversion 2.7% and complication 4.5%). CONCLUSION Preoperative knowledge of the increased conversion rate and increased morbidity will inform surgical planning for both the surgeon and the patient.


Plasma enzyme changes in carcinoma of bronchus mimicking myocardial infarction.


title: T cell responses to the structural polypeptides of porcine reproductive and respiratory syndrome virus.

text: The identification of antigens recognized by T cell responses has become fundamental for developing effective immunizations against viral infections. Lymphocyte proliferation and delayed-type hypersensitivity responses to porcine reproductive and respiratory syndrome virus (PRRSV) infection have been demonstrated. However, the polypeptide specificity of T cell responses to PRRSV is unknown. To identify the PRRSV polypeptides recognized by porcine lymphocytes two approaches were employed. First polypeptides of purified virions were separated by SDS-PAGE and particle suspensions obtained from nitrocellulose blots were used as antigens. Second, the polypeptides encoded by ORFs 2, 4, 5, 6, and 7 of the strain VR-2332 were expressed as fusion proteins with a histidine tag in mammalian cells, using vaccinia virus as expression system. Significant antigen-specific proliferation responses to the matrix and envelope proteins from purified virions were obtained. This finding was supported by specific and dose-dependent proliferation responses to the recombinant polypeptides encoded by ORF2, 5 and 6 detected in virus-infected but not in control pigs. These results demonstrate that T-cell responses can be detected to individual PRRSV polypeptides. The greater response to the product of ORF6 than to the other PRRSV polypeptides indicates that the viral matrix polypeptide may have a major role in cellular immunity.


The Common Cold.


title: Duodenal biopsies of HIV-infected patients with diarrhoea exhibit epithelial barrier defects but no active secretion.

text: OBJECTIVES To characterize diarrhoeal mechanisms in HIV-infected patients, epithelial transport and barrier function of the duodenal mucosa was investigated in vitro. PATIENTS Twenty-one HIV-seropositive patients (13 asymptomatic and eight with diarrhoea) and 12 controls from an urban referral-based tertiary care centre in Berlin who underwent duodenoscopy. METHODS A new miniaturized Ussing chamber allowed measurements on duodenal forceps biopsies. Epithelial barrier function was characterized by alternating current impedance analysis, which allows differentiation of epithelial and subepithelial resistance and by 3H-lactulose and 3H-mannitol flux measurements. Na+-glucose cotransport was quantified as phlorizin-sensitive short circuit current (Isc) and active ion secretion by baseline and bumetanide-sensitive Isc. RESULTS Duodenal biopsies from asymptomatic HIV-infected patients were no different from controls, whereas biopsies from HIV-infected patients with diarrhoea showed a decrease in epithelial resistance from 21.2+/-1.9 to 12.9+/-1.3 omega cm2 (P<0.01). Concomitantly, mucosal-to-serosal lactulose flux increased from 0.29+/-0.02 to 0.40+/-0.03 micromol (hcm2) (P<0.01). Phlorizin-sensitive Isc indicating Na+-glucose cotransport, as well as baseline and bumetanide-sensitive Isc indicating active electrogenic chloride secretion were not different between the three groups. CONCLUSIONS A miniaturized Ussing device was developed for electrophysiological investigations of duodenal forceps biopsies, which allowed characterization of active ion transport mechanisms and epithelial barrier function. Duodenum of HIV-infected patients with diarrhoea showed no evidence for active ion secretion or Na+-glucose malabsorption, but showed an impaired epithelial barrier function, which could contribute to diarrhoea by a leak flux mechanism.


title: Targeted disruption of the Ceacam1 (MHVR) gene leads to reduced susceptibility of mice to mouse hepatitis virus infection.

text: The CEACAM1 glycoproteins (formerly called biliary glycoproteins; BGP, C-CAM, CD66a, or MHVR) are members of the carcinoembryonic antigen family of cell adhesion molecules. In the mouse, splice variants of CEACAM1 have either two or four immunoglobulin (Ig) domains linked through a transmembrane domain to either a short or a long cytoplasmic tail. CEACAM1 has cell adhesion activity and acts as a signaling molecule, and long-tail isoforms inhibit the growth of colon and prostate tumor cells in rodents. CEACAM1 isoforms serve as receptors for several viral and bacterial pathogens, including the murine coronavirus mouse hepatitis virus (MHV) and Haemophilus influenzae, Neisseria gonorrhoeae, and Neisseria meningitidis in humans. To elucidate the mechanisms responsible for the many biological activities of CEACAM1, we modified the expression of the mouse Ceacam1 gene in vivo. Manipulation of the Ceacam1 gene in mouse embryonic stem cells that contained the Ceacam1a allele yielded a partial knockout. We obtained one line of mice in which the insert in the Ceacam1a gene had sustained a recombination event. This resulted in the markedly reduced expression of the two CEACAM1a isoforms with four Ig domains, whereas the expression of the two isoforms with two Ig domains was doubled relative to that in wild-type BALB/c (+/+) mice. Homozygous (p/p) Ceacam1a-targeted mice (Ceacam1aDelta4D) had no gross tissue abnormalities and were viable and fertile; however, they were more resistant to MHV A59 infection and death than normal (+/+) mice. Following intranasal inoculation with MHV A59, p/p mice developed markedly fewer and smaller lesions in the liver than +/+ or heterozygous (+/p) mice. The titers of virus produced in the livers were 50- to 100-fold lower in p/p mice than in +/p or +/+ mice. p/p mice survived a dose 100-fold higher than the lethal dose of virus for +/+ mice. +/p mice were intermediate between +/+ and p/p mice in susceptibility to liver damage, virus growth in liver, and susceptibility to killing by MHV. Ceacam1a-targeted mice provide a new model to study the effects of modulation of receptor expression on susceptibility to MHV infection in vivo.


A big push.


title: Sequence analysis and in vitro expression of genes 6 and 11 of an ovine group B rotavirus isolate, KB63: evidence for a non-defective, C-terminally truncated NSP1 and a phosphorylated NSP5.

text: An ovine group B rotavirus (GBR) isolate, KB63, was isolated from faeces of a young goat with diarrhoea in Xinjiang, People's Republic of China. Sequence determination and comparison of genes 6 and 11 with the corresponding sequences of GBR strains ADRV and IDIR showed that they were the cognate genes encoding NSP1 and NSP5, respectively. While the overall identities of nucleotide sequences between these two genes and the corresponding genes of strains ADRV and IDIR were in the range 52.6-57.2%, the identities of deduced amino acid sequences were only 34.9-46.3%. These results demonstrate that the substantial diversity of NSP1 observed among group A rotaviruses (GAR) also exists within GBRs and that a high degree of diversity also exists among NSP5 of GBRs, in contrast to GAR NSP5. The NSP1 gene of KB63 contains three ORFs, whereas the NSP1 genes of other GBR strains contain only two. ORFs 2 and 3 of the KB63 gene may be derived from a single ORF corresponding to ORF2 of other GBR strains by the usage of a stop codon created by an upstream single base deletion and single point mutations. In vitro expression studies showed that ORFs 1 and 2, but not 3, of gene 6 can be translated, suggesting that ORF2 may encode a C-terminally truncated, potentially functional product. It may play a role, together with the product of ORF1, in virus replication, as the virus can be passaged further in kids. Similarly, gene 11 can be translated in vitro. Like its counterpart in GARs, the protein encoded by gene 11 was shown to be phosphorylated in vitro.


title: Equine laparoscopy: gonadectomy.

text: Laparoscopic removal of the gonads from male and female horses is regularly performed for a variety of reasons. In female horses, common indications for ovariectomy include behavioral problems and signs of abdominal pain that are linked to estrus. Another indication is an abnormal, enlarged ovary, typically due to ovarian neoplasia, the most common being granulosa thecal cell tumor. Ovariectomy is also a treatment option for ovarian hematomas and other abnormalities. In addition, ovariectomy is used to sterilize mares intended for artificial insemination or embryo transfer programs.


title: [Over-expression in Escherichia coli and purification of nucleocaspid and membrane protein of SARS coronavirus].

text: Genes encoding nucleocaspid (N) and membrane (M) protein of SARS coronavirus were obtained by RT-PCR and were cloned into expression vector pET22b and pBV222. DNA sequencing showed that the genes cloned from a patient in Beijing were identical to the gene sequences from reported Toronto strain. The genes were over-expressed in E. coli either as inclusion body or as soluble form. The recombinant proteins were purified by ion-exchange, or ion-exchange followed by metal chelate affinity chromatography. The recombinant N protein was demonstrated highly antigenic and could be employed as antigen to detect SARS antibodies in ELISA system for SARS diagnosis.


title: Tunable dual-emitting shell-crosslinked nano-objects as single-component ratiometric pH-sensing materials.

text: Dual-emitting photonic nano-objects that can sense changes in the environmental pH are designed based on shell-crosslinked micelles assembled from amphiphilic block copolymers and crosslinked with pH-insensitive chromophores. The chromophoric crosslinkers are tetra-functionalized pyrazine molecules that bear a set of terminal aliphatic amine groups and a set of anilino amine groups, which demonstrate morphology-dependent reactivities towards the poly(acrylic acid) shell domain of the nano-objects. The extent to which the anilino amine groups react with the nano-object shell is shown to affect the hypsochromic shift (blue-shift). The ratio of fluorescence intensity at 496 nm over that of 560 nm is dependent upon the solution pH. We report, herein, observations on the pH-sensitive dual-emission photophysical properties of rod-shaped or spherical nano-objects, whose shell domains offer two distinct platforms for amidation reactions to occur-through formation of activated esters upon addition of carbodiimide or pre-installation of activated ester groups. We demonstrate that physical manipulations (changes in morphology or particle dimensions) or chemical manipulations of the crosslinking reaction (the order of installation of activated esters) lead to fine tuning of dual-emission over ca. 60 nm in a physiologically relevant pH range. Rod-shaped shell-crosslinked nanostructures with poly(p-hydroxystyrene) core show blue-shift as a function of increasing pH while spherical shell-crosslinked nanostructures with polystyrene core and poly(ethylene oxide) corona exhibit blue-shift as a function of decreasing pH.


title: 5th College of Physicians lecture -- a physician's odyssey: recollections and reflections.

text: Some reminiscences of events since the 1930s are presented chronologically. These include sketches of informal introduction to Medicine at the General Hospital (SGH), the amazing spirit of medical professionals during the "Syonan" years at Tan Tock Seng Hospital (TTSH) and of professors of Medicine and training in Hong Kong, Singapore and Britain. An account of practice of Medicine in the Civil Service, of teaching and research and of evolution of specialist training and examinations in the Academy of Medicine, the University's Post Graduate School (DGMS) and Specialist Accreditation Board (SAB) is given. The over-riding values of our profession are stressed, always contributing to improving the quality and standards of practice in the interests of our patients and fellow men.


title: Heptad repeat sequences are located adjacent to hydrophobic regions in several types of virus fusion glycoproteins.

text: Extensive regions of heptad repeat units consistent with an alpha-helical coiled coil conformation are located adjacent to hydrophobic, potentially fusion-related regions in the amino acid sequences of paramyxovirus fusion and retrovirus envelope glycoproteins. Similar arrangements of hydrophobic peptides and heptad repeat units exist in coronavirus peplomer proteins and influenza virus haemagglutinins. This suggests that there may be similarities in the structures of these proteins and in the functions of the hydrophobic fusion-related regions during virus entry.


title: [Single Incision Laparoscopic Ileocecal Resection for Small Intestinal Malignant Lymphoma - A Case Report].

text: A 76-year-old man with the chiefcomplaint ofmelena visited our hospital. A mass was palpable in the right lower abdomen. We diagnosed the illness as small intestinal malignant lymphoma in the terminal ileum and performed a single incision laparoscopic ileocecal resection "TANKO-ICR" with D3 lymph node dissection with the idea ofobtaining a wide visualization and surgical field. The Aesculap®internal organ retractor(B BRAUN)is very useful in such cases. We grasped the pedicle of the ileocecal artery using the retractor and pulled out the nylon line connected to the retractor by using ENDO CLOSETM (COVIDIEN)to extraabdomen for drawing the pedicle. Single incision laparoscopic surgery can be performed with only an umbilical wound; therefore, it is minimally invasive and has a superior aesthetic outcome. Small intestinal malignant lymphoma is a relatively rare malignant tumor ofthe digestive organs. We report this case demonstrating that single incision laparoscopic surgery was possible and provide a review ofthe relevant literature.


title: Interaction between a 54-kilodalton mammalian cell surface protein and cowpea mosaic virus.

text: Cowpea mosaic virus (CPMV), a plant virus that is a member of the picornavirus superfamily, is increasingly being used for nanotechnology applications, including material science, vascular imaging, vaccine development, and targeted drug delivery. For these applications, it is critical to understand the in vivo interactions of CPMV within the mammalian system. Although the bioavailability of CPMV in the mouse has been demonstrated, the specific interactions between CPMV and mammalian cells need to be characterized further. Here we demonstrate that although the host range for replication of CPMV is confined to plants, mammalian cells nevertheless bind and internalize CPMV in significant amounts. This binding is mediated by a conserved 54-kDa protein found on the plasma membranes of both human and murine cell lines. Studies using a deficient cell line, deglycosidases, and glycosylation inhibitors showed that the CPMV binding protein (CPMV-BP) is not glycosylated. A possible 47-kDa isoform of the CPMV-BP was also detected in the organelle and nuclear subcellular fraction prepared from murine fibroblasts. Further characterization of CPMV-BP is important to understand how CPMV is trafficked through the mammalian system and may shed light on how picornaviruses may have evolved between plant and animal hosts.


title: [Hematological changes and related gene mutation of post-severe acute respiratory syndrome patients with osteonecrosis].

text: OBJECTIVE To investigate the hematological changes and related gene mutation of post-severe acute respiratory syndrome (SARS) patients with osteonecrosis so as to find the sensitive molecular symbols for early screening of the high risk populations. METHODS Fast peripheral venous blood samples were collected from 61 post-SARS patients with osteonecrosis, 25 males and 36 females, aged 30.4 (20 - 60), and 52 sex and age-matched healthy persons as controls. ELISA was used to detect the coagulation and fibrinolysis indicators: activated partial thromboplastin time (APTT), protein C (PC), antithrombin III (AT-III), plasminogen activator inhibitor (PAI), activated protein C resistance (APC-R), plasminogen (PLG), von Willebrand factor (VWF), D-dimer (D-D), and fibrinogen (Fib). Real-time PCR was used to detect the mutation of factor V G1601A (FV Leiden) and prothrombin G20210A. RESULTS The levels of PC, AT-III, and PLG of the osteonecrosis group were 85% +/- 34%, 84 +/- 29%, and 69 +/- 23%, significantly lower than that of the control group (109% +/- 20%, 104% +/- 14%, and 94% +/- 15% respectively, all P < 0.01). PAI of the osteonecrosis group was 16 U/ml +/- 14 U/ml, significantly higher than that of the control group (8.0 U/ml +/- 4.3 U/ml, P < 0.01). The percentage of patients with abnormal indicators was 99.5% (54/61) in the osteonecrosis group, significantly higher than that of the control group (36.5%, 19/52, P < 0.01). The percentage of patients with 3 or more abnormal indicators was 72.1% (44/61) in the osteonecrosis group, significantly higher than that of the control group (17.3%, 9/52, P < 0.01). No mutations of F V Leiden and prothrombin G20210A was found in both groups. CONCLUSION Trends of hypercoagulation and hypofibrinolysis exist in the post-SARS patients with osteonecrosis. APTT, PC, AT-III, and PLG can be used as sensitive indicator for screening high risk populations of osteonecrosis.


title: [Receptor-binding ability of fragments 260-600 and 397-796 of SARS-associated coronavirus spike protein].

text: BACKGROUND To investigate the interaction between the host cell and the truncated S fragments to identify the receptor-binding domain of the spike (S) protein of SARS-associated coronavirus (SARS-CoV). METHODS Two different fragments S260-600 and S397-796 of the SARS-CoV S protein were expressed in Escherichia coli (E.coli) using a pET expression vector, respectively. The two recombinant proteins were separately verified by Western blot, purified by nickel-affinity chromatography, and incubated with Vero cells, a susceptible cell line of SARS-CoV infection, for cell binding assay. After the sequential probing with sera from convalescent SARS-patients and FITC-labeled anti-human IgG, the cells were analyzed by flow cytometry. The NIH 3T3 cell, a non-permissive cell line of SARS-CoV infection, was used as controls. RESULTS The recombinant proteins S260-600 and S397-796 were efficiently expressed in an insoluble form in E.coli. The appropriate expression of the proteins was confirmed by Western blotting using both SARS patients' sera and anti-6 x histidine antibody. The flow cytometry results showed that the both proteins were able to bind Vero cells, but the binding ability of S260-600 was somewhat stronger than that of S397-796. In contrast, the S260-600 protein did not bind NIH3T3 cells. CONCLUSION Both S260-600 and S397-796 exhibited different receptor binding activity. The S260-600 fragment probably contains the important receptor binding domain and could be a potential candidate for the development of SARS vaccine and anti-SARS therapeutics.


title: Vitamin D and Inflammation: Potential Implications for Severity of Covid-19.

text: Background Recent research has indicated that vitamin D may have immune supporting properties through modulation of both the adaptive and innate immune system through cytokines and regulation of cell signalling pathways. We hypothesize that vitamin D status may influence the severity of responses to Covid-19 and that the prevalence of vitamin D deficiency in Europe will be closely aligned to Covid-19 mortality. Methods We conducted a literature search on PubMed (no language restriction) of vitamin D status (for older adults) in countries/areas of Europe affected by Covid-19 infection. Countries were selected by severity of infection (high and low) and were limited to national surveys or where not available, to geographic areas within the country affected by infection. Covid-19 infection and mortality data was gathered from the World Health Organisation. Results Counter-intuitively, lower latitude and typically 'sunny' countries such as Spain and Italy (particularly Northern Italy), had low mean concentrations of 25(OH)D and high rates of vitamin D deficiency. These countries have also been experiencing the highest infection and death rates in Europe. The northern latitude countries (Norway, Finland, Sweden) which receive less UVB sunlight than Southern Europe, actually had much higher mean 25(OH)D concentrations, low levels of deficiency and for Norway and Finland, lower infection and death rates. The correlation between 25(OH)D concentration and mortality rate reached conventional significance (P=0.046) by Spearman's Rank Correlation. Conclusions Optimising vitamin D status to recommendations by national and international public health agencies will certainly have benefits for bone health and potential benefits for Covid-19. There is a strong plausible biological hypothesis and evolving epidemiological data supporting a role for vitamin D in Covid-19.


Multiple sclerosis and viruses.


title: The need for a national emergency health services database.

text: OBJECTIVE In February 2007, the Health Council of Canada, in its third annual report, emphasized the need for pan-Canadian data on our health care system. To date, no studies have examined the strengths and weaknesses of emergency health services (EHS) administrative databases, as perceived by researchers. We undertook a qualitative study to determine, from a researcher's perspective, the strengths and weaknesses of EHS administrative databases. The study also elicited researchers' suggestions to improve these databases. METHODS We conducted taped interviews with 4 Canadian health services researchers. The transcriptions were subsequently examined for common concepts, which were finalized after discussion with all the investigators. RESULTS Five common themes emerged from the interviews: clinical detail, data quality, data linkage, data use and population coverage. Data use and data linkages were considered strengths. Clinical detail, data quality and population coverage were considered weaknesses. CONCLUSION The 5 themes that emerged from this study all serve to reinforce the call from the Health Council of Canada for national data on emergency services, which could be readily captured through a national EHS administrative database. We feel that key stakeholders involved in emergency services across Canada should work together to develop a strategy to implement an accurate, clinically detailed, integrated and comprehensive national EHS database.


title: Clinical disease in children associated with newly described coronavirus subtypes.

text: OBJECTIVES Coronaviruses cause upper respiratory illness and occasionally lower tract disease in susceptible populations. In this study we examined the prevalence of 4 human coronaviruses, including subtypes OC43, 229E, and the recently described NL63 and HKU1 in a pediatric population presenting to a children's hospital. PATIENTS AND METHODS Specimens collected over a 1-year period from pediatric patients presenting with acute respiratory illness were analyzed for the presence of 4 coronavirus subtypes using consensus and subtype-specific real-time reverse-transcription polymerase chain reaction assays. The demographic and clinical characteristics associated with coronavirus infection were examined retrospectively. RESULTS Coronaviruses were detected in 66 of 1043 children. Eight, 11, 19, and 28 specimens were positive for subtypes 229E, NL63, OC43, and HKU1, respectively. Coronaviruses were detected throughout the study period; all 4 of the subtypes were present simultaneously in December. The acute clinical features were similar across subtypes. Of 32 children infected with a coronavirus as the sole respiratory pathogen, 13 had lower respiratory tract disease. Children whose only detectable respiratory virus was a coronavirus were more likely to have underlying chronic disease than were children coinfected with another respiratory virus. CONCLUSIONS Although 4 subtypes of coronavirus were detected, the recently discovered coronavirus subtypes NL63 and HKU1 accounted for the majority of coronaviruses detected in our cohort of mostly hospitalized children with respiratory symptoms. New subtypes likely represent a substantial portion of previously unexplained respiratory illnesses.


title: Characterization of a calf diarrheal coronavirus.

text: A coronavirus-like agent isolated from feces of a calf with diarrhea and attenuated by consecutive passage in a fetal bovine kidney cell line was characterized as a coronavirus. Negatively stained virions were approximately circular, had a mean diameter of 120 nm, and were covered with wide-spaced, petal-shaped projections about 20 nm long. Virions in ultrathin sections of infected cell monolayers had a mean diameter of 80 nm, lacked surface projections, and were found within cytoplasmic vesicles. Viral antigen was demonstrated by immunofluorescence microscopy to occur only in cytoplasm. Growth of the virus was not inhibited by 5-iodo-2'- deoxyuridine and actinomycin D. The virus was sensitive to ether, chloroform, deoxycholate, and heat treatment. However, thermosensitivity was stabilized in the presence of 1 M MgCl2; at pH 3, the virus was stable. Hemadsorption and hemagglutination were observed with erythrocytes of hamsters, mice, and rats but not with erythrocytes of cats, dogs, goats, sheep, cattle, horses, turkeys, chickens, guinea pigs, rabbits, geese, pigs, and man (type O). However, hemadsorption and hemagglutination were shown to be virus specific, since this could be inhibited by specific antiserum. Both infectivity and hemagglutinating activity were maximal at a particle density of 1.18 g/ml by sucrose density gradient centrifugation, indicating that hemagglutinin was part of the virion.


title: Antiapoptotic BCL-2 is required for maintenance of a model leukemia.

text: Resistance to apoptosis, often achieved by the overexpression of antiapoptotic proteins, is common and perhaps required in the genesis of cancer. However, it remains uncertain whether apoptotic defects are essential for tumor maintenance. To test this, we generated mice expressing a conditional BCL-2 gene and constitutive c-myc that develop lymphoblastic leukemia. Eliminating BCL-2 yielded rapid loss of leukemic cells and significantly prolonged survival, formally validating BCL-2 as a rational target for cancer therapy. Loss of this single molecule resulted in cell death, despite or perhaps attributable to the presence of other oncogenic events. This suggests a generalizable model in which aberrations inherent to cancer generate tonic death signals that would otherwise kill the cell if not opposed by a requisite apoptotic defect(s).


title: SARS: a new infectious disease for a new century.

text: BACKGROUND A new infectious disease swept the world in early 2003, causing restrictions on international travel and economic downturn in affected countries. OBJECTIVE This article aims to provide an overview of the epidemic of severe acute respiratory syndrome (SARS). DISCUSSION SARS is caused by a new coronavirus thought to be of animal origin. The virus induces symptoms of atypical pneumonia, clinically indistinguishable from similar syndromes. The severity is such that a 15% mortality rate has been reported. No treatment has yet been identified as reliably successful. Transmission is by droplet spread, requiring close contact. Stringent infection control precautions in health care institutions, broad isolation measures in affected communities and international surveillance with barrier restrictions to travel have led to termination of the epidemic. As of July 11, 2003, 8437 people in 32 countries have been affected, with 813 deaths reported.


title: [Coronavirus and "astrovirus" observed in stools of children with gastroenteritis (author's transl)].

text: In a year's survey (December 1976-November 1977) of 190 children admitted to hospital with acute gastroenteritis, an "aetiological" agent was found in stools by electron microscopy in 41% of cases. Rotavirus were found in 34% of patients. We also found Coronavirus-like particles and Astrovirus-like particles in 25 and 5 stools, respectively. Improvements are awaited to demonstrate the responsibility of these virus in gastroenteritis.


title: Interferon induction in rotavirus and coronavirus infections: a review of recent results.

text: The interferon (IFN) response was studied in two different models of viral enteritis of the neonate. In rotavirus infected calves, IFN synthesis could be detected in the intestine and in the blood at the time of the diarrheic symptoms. A kinetic study on canulated calves suggested that systemic IFN is of intestinal origin. During TGEV infection (Transmissible gastroenteritis virus) of the newborn piglet, an acute disease which leads to 100% mortality, IFN was found at very high titres (1 000-20 000 u/ml) in the intestine, blood, urine and other organs. Intestinal IFN synthesis started some hours after the onset of diarrhea and was very transient, i.e. no more detectable two days p.i. Unlike the calf situation, IFN response in the serum lasted much longer suggesting an extra-intestinal origin. As a confirmation, piglets infected with cell-adapted strains had high levels of circulating IFN before the onset of intestinal IFN and of diarrhea. Virus and IFN were found in the lungs, due to a so far unrecognized tropism of TGEV for the macrophages. These findings indicate that the pathogenesis of TGEV is more complex than previously claimed.


title: Effects of ambient temperature on volume, specialty composition and triage levels of emergency department visits.

text: AIM To evaluate the effects of change of ambient temperature on emergency department (ED) patient visits. METHODS This prospective observational study was conducted in the ED of National Taiwan University Hospital from January 2002 to January 2007. The daily ED patient numbers of different triage levels in different service specialties were collected and correlated with the daily average temperature (T) and change in temperature (deltaT) compared with the previous day. A univariate analysis was performed with the Pearson correlation and a multivariate analysis with multiple linear regression analysis. RESULTS A total of 505,224 patient visits were included in this study. On univariate analysis, there was no significant correlation between T and the ED volume (r = 0.012, p = 0.608), but there was a significant correlation between deltaT and ED volume (r = 0.109, p<0.001). On multivariate analysis, deltaT and holidays were identified as independent predictors of ED volume. We established the following formula in predicting the ED patient number: y = 265.42 + (0.06 x T) + (2.57 x deltaT) + (59.77 x holiday). There was a positive association between T and the trauma patient number, while there was a negative association between T and medical and paediatric patient numbers. On the triage level, a low T was associated with increased patient triage level, while no significant association was noted between deltaT and the proportion in any triage level. CONCLUSIONS Our study demonstrated that ambient temperature had differential effects on ED patient visits of different specialties and severities.


Ripped from the headlines: how can we harness communications to control TB?


title: RNA signals in the 3' terminus of the genome of Equine arteritis virus are required for viral RNA synthesis.

text: RNA virus genomes contain cis-acting sequences and structural elements involved in virus replication. Both full-length and subgenomic negative-strand RNA synthesis are initiated at the 3' terminus of the positive-strand genomic RNA of Equine arteritis virus (EAV). To investigate the molecular mechanism of EAV RNA synthesis, the RNA secondary structure of the 3'-proximal region of the genome was analysed by chemical and enzymic probing. Based on the RNA secondary structure model derived from this analysis, several deletions were engineered in a full-length cDNA copy of the viral genome. Two RNA domains were identified that are essential for virus replication and most likely play a key role in viral RNA synthesis. The first domain, located directly upstream of the 3' untranslated region (UTR) (nt 12610-12654 of the genome), is mainly single-stranded but contains one small stem-loop structure. The second domain is located within the 3' UTR (nt 12661-12690) and folds into a prominent stem-loop structure with a large loop region. The location of this stem-loop structure near the 3' terminus of the genome suggests that it may act as a recognition signal during the initiation of minus-strand RNA synthesis.


title: Action of disinfectants on canine coronavirus replication in vitro.

text: Canine coronavirus (CCoV) is responsible for enteric disease in pups. Infected dogs generally have a rapid recovery, so the virus is highly contagious and the spread of infection is difficult to control. Chemical disinfectants have been widely used in human disease-control programmes to prevent viral infectious diseases from spreading, but to date, there are no studies in the literature on the sensitivity of CCoV to chemical biocides. The present study investigated the sensitivity of CCoV to disinfectants currently used for prophylaxis in kennel and dog breeding locations. The effects of three agents: alkyl-dimethyl-benzyl-ammonium chloride, benzalkonium chloride and didecyl-dimethyl-ammonium chloride, on the infectivity titre of CCoV in A72 cell lines, were studied at different concentrations. Although they may regard a small number of agents, the findings showed that the sensitivity of CCoV to disinfectants varies and the differences are dose correlated. In general, virus inactivation implies a permanent loss of infectivity which can be evaluated in suspensions and hand disinfection tests.


title: Cloning and sequencing of the gene encoding the spike protein of the coronavirus IBV.

text: RNA sequences encoding the surface projection (spike) of the coronavirus infectious bronchitis virus, strain Beaudette, have been cloned into pBR322 using cDNA primed with a specific oligonucleotide. A 5.3 kilobase viral insert in the clone pMB179 has been identified. The region of this clone coding for the spike gene has been sequenced by the chain termination method, and we present here the first report of DNA sequence data for a coronavirus spike protein, the protein which forms the characteristic 'corona' after which the group is named. The amino acid sequence of the primary translation product, deduced from the DNA sequence, predicts a polypeptide of 1162 amino acids with a molecular weight of 127 006. This has many interesting features which confirm and extend our knowledge of this recently characterized membrane glycoprotein. The polypeptide is subsequently cleaved to S1 and S2, and partial amino acid analysis of the amino-terminus of the S1 polypeptide has been employed to locate the position of this terminus of S1 within the large open reading frame. The amino acid analysis also reveals the presence of an 18 amino acid putative signal sequence on the primary translation product which is not present on the mature S1 polypeptide.


title: Laparoscopic Sleeve Gastrectomy Outcomes of 750 Patients: A 2.5-Year Experience at a Bariatric Center of Excellence.

text: Laparoscopic sleeve gastrectomy (LSG) is gaining popularity worldwide. This retrospective cohort study evaluated the outcomes of a large cohort of patients with obesity who underwent LSG in a Bariatric Center of Excellence. All consecutive patients who underwent LSG between July 2013 and April 2016 were identified retrospectively. Preoperative and postoperative variables and comorbidities were recorded. The study consisted of 750 patients. Their mean age was 37.4 years; 72% were women, and the mean body mass index was 42.8 kg/m. The most common preoperative comorbidities were diabetes (23.3%), hyperlipidemia (21.9%), hypertension (21.1%), and obstructive sleep apnea (21.1%). The rates of comorbidity resolution during follow-up were 80.6%, 74.4%, 82.9%, and 94.3%, respectively. The percentage average excess weight loss 1, 3, and 6 months and 1 and 2 years after surgery was 29.4%±11.3%, 54.4%±17.7%, 76.9%±20.9%, 85.5%±23.6%, and 89.7%±27.6%, respectively. There was no mortality. LSG effectively and safely induced weight loss and comorbidity resolution.


title: [Acute gastroenteritis of viral etiology].

text: Retrospective analysis of own experiences and investigations concerning the role of the viral etiology in acute gastroenteritis were presented. Stools of children and soldiers with acute gastroenteritis were examined by electron microscopy. The most frequent causative agents were rotaviruses and much less frequently found were other viral particles of: coronaviruses, adenoviruses, Norwalk-like viruses, astroviruses, caliciviruses and "small round viruses".


ASCOT-LLA: questions about the benefits of atorvastatin.


docid: eaergjhc

title: The S2 subunit of the murine coronavirus spike protein is not involved in receptor binding.

text: The receptor-binding capacity of the S2 subunit of the murine coronavirus S protein was examined by testing the inhibition of virus-receptor binding. Sp-4 virus and S1N(330), which consists of the N-terminal 330 amino acids of the S1 protein, both of which exhibited receptor-binding capacity, were able to prevent the binding of cl-2 virus to the receptor, while the mutant protein S1N(330)-159, which failed to bind to the receptor protein, and S2TM-, which lacks the transmembrane and cytoplasmic domains normally existing in the S2, were unable to prevent the binding of cl-2. By using cultured DBT cells, it was revealed that the infection of cells by cl-2 virus was significantly inhibited by S1N(330) but not by S2TM-. These results indicate that the S2 protein is not involved in the receptor binding of murine coronaviruses.


ABNM: Helping Diplomates and Trainees During the COVID-19 Pandemic.


title: Description and clinical treatment of an early outbreak of severe acute respiratory syndrome (SARS) in Guangzhou, PR China.

text: Severe acute respiratory syndrome (SARS), now known to be caused by a coronavirus, probably originated in Guangdong province in southern China in late 2002. The first major outbreak occurred in Guangzhou, the capital of Guangdong, between January and March 2003. This study reviews the clinical presentation, laboratory findings and response to four different treatment protocols. Case notes and laboratory findings were analysed and outcome measures were collected prospectively. The SARS outbreak in Guangdong province and the outbreak in Guangzhou associated with hospitals in the city are described, documenting clinical and laboratory features in a cohort of 190 patients randomly allocated to four treatment regimens. Patients were infected by close contact in either family or health-care settings, particularly following procedures likely to generate aerosols of respiratory secretions (e.g. administration of nebulized drugs and bronchoscopy). The earliest symptom was a high fever followed, in most patients, by dyspnoea, cough and myalgia, with 24 % of patients complaining of diarrhoea. The most frequent chest X-ray changes were patchy consolidation with progression to bilateral bronchopneumonia over 5-10 days. Thirty-six cases developed adult respiratory distress syndrome (ARDS), of whom 11 died. There was no response to antibiotics. The best response (no deaths) was seen in the group of 60 patients receiving early high-dose steroids and nasal CPAP (continuous airway positive pressure) ventilation; the other three treatment groups had significant mortality. Cross-infection to medical and nursing staff was completely prevented in one hospital by rigid adherence to barrier precautions during contact with infected patients. The use of rapid case identification and quarantine has controlled the outbreak in Guangzhou, in which more than 350 patients have been infected. Early administration of high-dose steroids and CPAP ventilation appears to offer the best supportive treatment with a reduced mortality compared with other treatment regimens.


title: Introduction of microsurgery in Vietnam by a charitable organization: a 15-year experience.

text: BACKGROUND Microsurgical procedures, although equipment- and labor-intensive, allow efficient treatment of selected soft-tissue, bone, and peripheral nerve defects. The precise surgical skills required and the high equipment and institutional costs have been deterrents to initiating programs in developing countries. The authors report their 15-year international effort in facilitating the development of microsurgical techniques in Vietnam. METHODS The authors reviewed their educational, logistical, and operative experience from 11 Operation Smile International missions to Vietnam and the microsurgical procedures performed independently by Vietnamese surgeons at the Central Military Hospital 108 in Hanoi. RESULTS Over 15 years, Operation Smile International missions to Vietnam performed 108 free tissue transfer operations with 15 peripheral nerve transfer procedures and 143 nonmicrosurgical reconstructive operations. Visiting surgeons with specialized expertise taught facial reanimation, flap prefabrication, and perforator flaps. During this same period, Vietnamese surgeons became facile with microsurgical techniques and independently performed a wide array of these procedures in the institutions visited. Vietnamese surgeons have organized microsurgery divisions within some hospital departments and now teach microsurgical techniques. Repeated missions allowed for patient follow-up, staged procedures, educational exchange, and quality control. Several Vietnamese surgeons have traveled abroad to obtain additional training and have set up training programs in other areas of Vietnam. CONCLUSIONS Charitable organizations can help surgeons in developing countries master complex microsurgical techniques through short-term medical missions, donation of equipment and supplies, and the encouragement of institutional support. A continuing education program, including local conferences, microsurgical laboratory training facilities, and study abroad, can aid this introduction.


title: Regulation of closterovirus gene expression examined by insertion of a self-processing reporter and by northern hybridization.

text: A reporter open reading frame (ORF) coding for a fusion of bacterial beta-glucuronidase (GUS) with a proteinase domain (Pro) derived from tobacco etch potyvirus was utilized for tagging individual genes of beet yellows closterovirus (BYV). Insertion of this reporter ORF between the first and second codons of the BYV ORFs encoding the HSP70 homolog (HSP70h), a major capsid protein (CP), and a 20-kDa protein (p20) resulted in the expression of the processed GUS-Pro reporter from corresponding subgenomic RNAs. The high sensitivity of GUS assays permitted temporal analysis of reporter accumulation, revealing early expression from the HSP70h promoter, followed by the CP promoter and later the p20 promoter. The kinetics of transcription of the remaining BYV genes encoding a 64-kDa protein (p64), a minor capsid protein (CPm), and a 21-kDa protein (p21) were examined via Northern blot analysis. Taken together, the data indicated that the temporal regulation of BYV gene expression includes early (HSP70h, CPm, CP, and p21 promoters) and late (p64 and p20 promoters) phases. It was also demonstrated that the deletion of six viral genes that are nonessential for RNA amplification resulted in a dramatic increase in the level of transcription from one of the two remaining subgenomic promoters. Comparison with other positive-strand RNA viruses producing multiple subgenomic RNAs showed the uniqueness of the pattern of closterovirus transcriptional regulation.


Chimeras in the crosshairs.


Infection prevention and control guidelines for patients with Middle East Respiratory Syndrome Coronavirus (MERS-CoV) infection.


title: The Nucleocapsid Protein of Coronaviruses Acts as a Viral Suppressor of RNA Silencing in Mammalian Cells.

text: RNA interference (RNAi) is a process of eukaryotic posttranscriptional gene silencing that functions in antiviral immunity in plants, nematodes, and insects. However, recent studies provided strong supports that RNAi also plays a role in antiviral mechanism in mammalian cells. To combat RNAi-mediated antiviral responses, many viruses encode viral suppressors of RNA silencing (VSR) to facilitate their replication. VSRs have been widely studied for plant and insect viruses, but only a few have been defined for mammalian viruses currently. We identified a novel VSR from coronaviruses, a group of medically important mammalian viruses including Severe acute respiratory syndrome coronavirus (SARS-CoV), and showed that the nucleocapsid protein (N protein) of coronaviruses suppresses RNAi triggered by either short hairpin RNAs or small interfering RNAs in mammalian cells. Mouse hepatitis virus (MHV) is closely related to SARS-CoV in the family Coronaviridae and was used as a coronavirus replication model. The replication of MHV increased when the N proteins were expressed in trans, while knockdown of Dicer1 or Ago2 transcripts facilitated the MHV replication in mammalian cells. These results support the hypothesis that RNAi is a part of the antiviral immunity responses in mammalian cells. IMPORTANCE RNAi has been well known to play important antiviral roles from plants to invertebrates. However, recent studies provided strong supports that RNAi is also involved in antiviral response in mammalian cells. An important indication for RNAi-mediated antiviral activity in mammals is the fact that a number of mammalian viruses encode potent suppressors of RNA silencing. Our results demonstrate that coronavirus N protein could function as a VSR through its double-stranded RNA binding activity. Mutational analysis of N protein allowed us to find out the critical residues for the VSR activity. Using the MHV-A59 as the coronavirus replication model, we showed that ectopic expression of SARS-CoV N protein could promote MHV replication in RNAi-active cells but not in RNAi-depleted cells. These results indicate that coronaviruses encode a VSR that functions in the replication cycle and provide further evidence to support that RNAi-mediated antiviral response exists in mammalian cells.


title: Cost Differences Between Open and Minimally Invasive Surgery.

text: PURPOSE To analyze the cost difference between minimally invasive surgery (MIS) and open surgery from a commercial payer perspective for colectomy, ventral hernia repair, thoracic resection (resection of the lung), and hysterectomy. DESIGN A retrospective claims data analysis was conducted using the 2011 and 2012 Truven Health Analytics MarketScan Commercial Claims and Encounter Database. Study eligibility criteria included age 18-64 years, pharmacy coverage, ≥ 1 month of eligibility in 2012, and a claim coded with 1 of the 4 surgical procedures of interest; the index year was 2012. METHODOLOGY Average allowed facility and professional costs were calculated during inpatient stay (or day of surgery for outpatient hysterectomy) and the 30 days after discharge for MIS vs open surgery. Cost difference was compared after adjusting for presence of cancer, geographic region, and risk profile (age, gender, and comorbidities). RESULTS In total, 46,386 cases in the 2012 MarketScan database represented one of the surgeries of interest. The difference in average allowed surgical procedure cost (facility and professional) between open surgery vs adjusted MIS was $10,204 for colectomy; $3,721, ventral hernia repair; $12,989, thoracic resection; and $1,174, noncancer hysterectomy (P < .001 for all comparisons).The difference in average allowed cost in the 30 days after surgery between open surgery vs adjusted MIS was $1,494 for colectomy, $1,320 for ventral hernia repair, negative $711 for thoracic resection, and negative $425 for noncancer hysterectomy (P < .001 for all comparisons, except P = .487 for thoracic resection). CONCLUSION MIS was associated with statistically significantly lower costs than open surgery for all 4 analyzed surgeries.


title: A personalist approach to public-health ethics.

text: First we give an overview of the historical development of public health. Then we present some public-health deontology codes and some ethical principles. We highlight difficulties in defining ethics for public health, with specific reference to three of them that concern: (i) the adaptability to public health of the classical principles of bioethics; (ii) the duty to respect and safeguard the individual while acting within the community perspective that is typical of public health; and (iii) the application-oriented nature of public health and the general lack of attention towards the ethical implications of collective interventions (compared with research). We then mention some proposals drafted from North American bioethics "principles" and utilitarian, liberal and communitarian views. Drawing from other approaches, personalism is outlined as being the theory that offers a consistent set of values and alternative principles that are relevant for public health.


title: Climate change and animal diseases in South America.

text: Climate strongly affects agriculture and livestock production and influences animal diseases, vectors and pathogens, and their habitat. Global warming trends predicted in the 2007 Intergovernmental Panel on Climatic Change (IPCC) report for South America are likely to change the temporal and geographical distribution of infectious diseases, including those that are vector-borne such as bluetongue, West Nile fever, vesicular stomatitis and New World screwworm. Changes in distribution will be partially modulated by El Niño Southern Oscillation events, which will become more frequent and lead to a greater frequency of droughts and floods. Active disease surveillance for animal diseases in South America, particularly for vector-borne diseases, is very poor. Disease reporting is often lacking, which affects knowledge of disease distribution and impact, and preparedness for early response. Improved reporting for animal diseases that may be affected by climate change is needed for better prevention and intervention measures in susceptible livestock, wildlife and vectors in South America. This requires contributions from multidisciplinary experts, including meteorologists, epidemiologists, biologists and ecologists, and from local communities.


title: Production, characterization, and uses of monoclonal antibodies against recombinant nucleoprotein of elk coronavirus.

text: This is the first report of the production of monoclonal antibodies against elk coronavirus. The nucleoprotein gene of elk coronavirus was amplified by PCR and was cloned and expressed in a prokaryotic expression vector. Recombinant nucleocapsid protein was used to immunize mice for the production of hybridomas. Twelve hybridomas that produced monoclonal antibodies against the nucleocapsid protein of elk coronavirus were selected by an indirect fluorescent-antibody test, an enzyme-linked immunosorbent assay, and a Western blot assay. Ten of the monoclonal antibodies were of the immunoglobulin G1 (IgG1) isotype, one was IgG2a, and one was IgM. All had kappa light chains. By immunohistochemistry four monoclonal antibodies detected bovine coronavirus and elk coronavirus in formalin-fixed intestinal tissues. Antinucleoprotein monoclonal antibodies were found to be better at ruminant coronavirus detection than the anti-spike protein monoclonal antibodies. Because nucleoprotein is a more abundant antigen than spike protein in infected cells, this was not an unexpected finding.


title: A Pathophysiological Perspective on COVID-19's Lethal Complication: From Viremia to Hypersensitivity Pneumonitis-like Immune Dysregulation.

text: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the coronavirus responsible for our recent coronavirus disease 2019 pandemic, is driving a lung immunopathology that strongly resembles a severe form of hypersensitivity pneumonitis (HP). A review of recent Severe acute respiratory syndrome-related coronavirus (SARS-CoV) and SARS-CoV-2 medical reports, as well as described characteristics of HP, lead us to postulate a theory for SARS-CoV-2 severe disease. We propose that the novel SARS-CoV-2 can act as a trigger and substrate of an HP-like severe immune reaction especially in genetically vulnerable individuals in addition to those with immune senescence and dysregulation. Accordingly, the purpose of our letter is to shift the emphasis of concern surrounding immune activity from viral infection to an HP-like severe immune reaction. We review similarities in disease presentation between infection and allergy, relevant immunopathology, and outline phases of SARS-CoV-2 disease with perspectives on therapy and critical care. Altogether, the favored course is to begin treatments that address the disease at the earliest phase before immune dysregulation leading to uncontrolled pulmonary inflammation.


title: Clinical research in a public health crisis: the integrative approach to managing uncertainty and mitigating conflict.

text: In order to advance science while preserving social solidarity and institutional trust, clinical research must carefully manage ethical tensions created by the two overlapping dynamics of conflict and uncertainty. One of these dynamics is inherent in the research enterprise itself and the other arises in the particular context of a public health emergency. One test for both the moral soundness and practical utility of a framework for research ethics is its ability to help stakeholders understand and manage these ethical tensions as much as possible. After clarifying the dynamics that give rise to these tensions, this paper argues that two common approaches to evaluating clinical research have significant shortcomings in this regard. This paper then sketches and defends the integrative approach to managing these tensions.


docid: gzcwbvjm

title: Influence of a new monofilament polyester mesh on inflammation and matrix remodeling.

text: ABSTRACT Synthetic mesh is widely used for hernia repairs, but mesh-induced chronic inflammatory responses may lead to postoperative complications. We previously showed an elevated response to multifilament polyester (PE) versus monofilament polypropylene (PP) and polytetrafluoroethylene (PTFE) meshes, but it is unclear whether this discrepancy is due to the differences in chemical composition or filament structure. This study compares the influence of a newly available monofilament PE mesh to that of multifilament PE, monofilament PP, and monofilament PTFE on the expression of genes important in inflammation and extracellular matrix remodeling in a rat model. Full thickness abdominal wall defects were corrected with onlay repair or suture repair with no mesh. Explants were harvested 7 or 90 days after repair and divided for histology and mRNA analyses using real-time quantitative polymerase chain reaction arrays to profile expression at the tissue-mesh interface. Monofilament PE elicited a reduced foreign body reaction compared to multifilament PE, corresponding with reduced mRNA expression of important inflammatory cytokines and matrix metalloproteinases (MMPs). Unexpectedly, monofilament PE also resulted in markedly reduced mRNA expression of tumor necrosis factor and MMPs 3 and 9 compared to the widely-used monofilament PP mesh. Findings from this study revealed that both chemical composition and filament structure are important mesh characteristics that may affect a patient's wound healing response and clinical outcome, and should be considered by the surgeon when choosing a particular mesh. Although clinical studies are warranted, results in a rodent model suggest that monofilament PE may be more beneficial than the multifilament form for certain hernia repairs.


title: Neonatal pertussis diagnosis: low procalcitonin level and high lymphocyte count are able to discriminate pertussis from bacterial and viral infections.

text: Pertussis is quite frequent and severe among infants; therefore, rapid diagnosis and timely targeted therapy are essential. Although a molecular test for etiological diagnosis is now available, it may not be available everywhere, and therefore adjunctive diagnostic tests are still useful for presumptive diagnosis. We describe the use of procalcitonin (PCT) and lymphocyte count to discriminate among pertussis, bacterial and viral infections. Fourteen infants per group were studied. The decision tree, built considering all available variables, showed a major role of PCT in predicting the different groups. A PCT value equal to or greater than 0.75 ng/ml selected for bacterial infections. A PCT value lower than 0.75 ng/ml and a lymphocyte count equal to or greater than 10,400/mm3 selected the subjects with pertussis, while a lymphocyte count lower than 10,400/mm3 selected for viral etiology. PCT should be used in the diagnosis of infants suspected of having pertussis.


title: [High-isolation preparedness for very contagious and potential severely ill patients].

text: Ten and seven years ago respectively, the two high-isolation units in Denmark became prepared to receive highly contagious and potential severely ill patients. The units are located in the departments of infectious diseases in Hvidovre Hospital and Aarhus University Hospital. Both departments had little experience until the Ebola outbreak in West Africa, 2013-2015. Altogether, 15 possible Ebola patients were examined in the two departments, but the diagnosis was ruled out for all of them. Besides, a few possible candidates for Middle East respiratory syndrome patients were examined. Always being prepared in a busy clinical everyday environment is a challenge, and it takes regular training and exercises involving all personnel.


title: HMGB1-induced endothelial cell pyroptosis is involved in systemic inflammatory response syndrome following radiofrequency ablation of hepatic hemangiomas.

text: The goal of this study was to assess the role of high-mobility group box 1 (HMGB1)-induced endothelial cell (EC) pyroptosis in systemic inflammatory response syndrome (SIRS) following radiofrequency (RF) ablation of hepatic hemangiomas. We enrolled 76 patients with hepatic hemangioma who underwent RF ablation. Serum concentrations of HMGB1, interleukin (IL)-1β, IL-18 and lactate dehydrogenase (LDH) were determined at different time points. Immunohistochemistry staining (IHC) was performed to evaluate the expressions of HMGB1, NLRP3, caspase-1, GSDMD, IL-18 and IL-1β in hepatic hemangioma and sub-ablated hemangioma tissues. In vitro experiments used human umbilical vein endothelial cells (HUVECs) treated with sub-ablative hyperthermia to mimic insufficient RF ablation of hepatic hemangiomas. ELISA and western blotting were performed to quantify HMGB1, NLRP3, caspase-1, GSDMD, IL-18, IL-1β and LDH levels with or without the addition of ethyl pyruvate (EP), a HMGB1 inhibitor, in the medium. Flow cytometry and fluorescent staining were performed to assess pyroptosis of HUVECs. Twenty-nine patients experienced SIRS after RF ablation (29/76, 38.2%). HMGB1, IL-1β and IL-18 levels were significantly correlated with SIRS. IHC staining revealed an obvious increase in HMGB1, NLRP3, caspase-1, GSDMD, IL-18, and IL-1β in the ECs of sub-ablated hemangioma but not in hepatic hemangioma. In vitro experiments showed that subablative hyperthermia led to HMGB1-induced pyroptosis of HUVECs and EP attenuated the pyroptosis of HUVECs. Taken together, these data demonstrate HMGB1-induced ECs pyroptosis may occur during SIRS following RF ablation of hepatic hemangiomas.


title: Correlation between neonatal calf diarrhea and the level of maternally derived antibodies.

text: Passively acquired antibodies through colostrum will protect calves against etiological agents of neonatal calf diarrhea. Among them enteric diseases due to strains of Enterotoxigenic Escherichia coli (ETEC) are the most commonly occurring form of colibacillosis in newborn calves. Specific antibodies against whole ETEC cells and total immunoglobulin G in dam serum, colostrum and calf serum were determined. There were significant differences (P=0.0005) between antibody titers in normal and diarrheic groups, in which diarrheic group had a higher titer. Total IgG concentration in diarrheic calves (20.86 ± 0.49), their dams (23.48 ± 0.54) and colostrum (33.40 ± 0.50) was less than normal group (P=0.0005). There was a highly significant positive correlation between dam total IgG with calf total IgG (r=0.022; ratio=52.11). Colostral anti-E. coli antibody had a highly significant positive correlation with anti-E. coli in calf serum (r=0.345; ratio=0.62). Anti-E. coli antibody in calf serum had a highly significant negative correlati with total IgG of dam serum, colostrum and calf serum. While the level of anti-E. coli antibodies in diarrheic group was considerably higher than normal group, our findings reported here are in agreement that immunity to diarrhea also might be correlated with maternal cells or cellular components as well as cytokines which are transferred by colostrum to neonatal calves. Nevertheless, the level of maternally derived antibodies is a promising indicator for passive immunity and protection against diarrhea in neonatal calves.


title: Three intergenic regions of coronavirus mouse hepatitis virus strain A59 genome RNA contain a common nucleotide sequence that is homologous to the 3' end of the viral mRNA leader sequence.

text: cDNA clones that represent various portions of the coronavirus mouse hepatitis virus strain A59 genome RNA have been constructed. cDNAs were synthesized by transcription of genome RNA by using either oligo(dT) or random oligomers of calf thymus DNA as primers. These cDNAs were converted into double-stranded DNA and cloned into pBR322 by standard techniques. The resulting cloned viral DNA fragments were mapped to viral genes by hybridization with Northern blots of intracellular RNA from mouse hepatitis virus strain A59-infected cells. These cDNA clones map in six of the seven viral genes. Clone g344, 1.8 kilobases, is the largest and encompasses gene 5 (which encodes a nonstructural protein) and gene 6 (which encodes the E1 viral glycoprotein) as well as the intergenic regions preceding genes 5, 6, and 7. Sequencing of parts of this cloned DNA show that these three intergenic regions contain a common 11-nucleotide sequence. This sequence shares homology with the 3' end of the viral mRNA leader sequence. Thus, this common intergenic sequence may contain a binding site for a leader RNA that hybridizes to negative-strand viral RNA at the beginning of each gene to prime mRNA synthesis. The different degrees of homology between the leader and its putative binding site may influence the differential rates of transcription of the various viral mRNAs.


title: Homologous RNA recombination allows efficient introduction of site-specific mutations into the genome of coronavirus MHV-A59 via synthetic co-replicating RNAs.

text: We describe a novel strategy to site-specifically mutagenize the genome of an RNA virus by exploiting homologous RNA recombination between synthetic defective interfering (DI) RNA and the viral RNA. Marker mutations introduced in the DI RNA were replaced by the wild-type residues during replication. More importantly, however, these genetic markers were introduced into the viral genome: even in the absence of positive selection MHV recombinants could be isolated. This finding provides new prospects for the study of coronavirus replication using recombinant DNA techniques. As a first application, we describe the rescue of the temperature sensitive mutant MHV Albany-4 using DI-directed mutagenesis. Possibilities and limitations of this strategy are discussed.


title: Protection of dogs against canine distemper by vaccination with a canarypox virus recombinant expressing canine distemper virus fusion and hemagglutinin glycoproteins.

text: OBJECTIVES To evaluate the safety and efficacy of a live canarypox virus recombinant-canine distemper virus (CDV) combination vaccine against virulent CDV challenge exposure, and to document lack of interference among the other modified-live virus (MLV) components. ANIMALS 33 specific-pathogen-free (SPF) Beagle pups (7 to 10 weeks old). PROCEDURE A canarypox virus recombinant-CDV combination vaccine was tested for safety and efficacy along with MLV components (canine adenovirus type 2, canine coronavirus, canine parainfluenza virus, and canine parvovirus) in 26 SPF Beagle pups. The combination vaccine was rehydrated with either Leptospira canicola-L icterohaemorrhagiae combination bacterin (vaccine 1) or sterile diluent (vaccine 2). An additional group of 7 seronegative SPF pups received the control MLV components devoid of the combination vaccine (vaccine 3). Two vaccinations were administered 21 days apart, either IM or SC. The dose of the combination vaccine used to inoculate these pups was 40 times lower than the recommended commercial dose. At 21 days after the booster vaccination, all pups were challenge exposed with a virulent CDV strain, then were observed for 21 days to record morbidity and mortality. RESULTS Adverse local or generalized reactions were not induced by vaccinations. All vaccinates seroconverted to CDV. Serum antibody titers to MLV components were not different, with or without inclusion of the combination vaccine. After challenge exposure, morbidity and mortality in vaccinates were 0% (0/26); in control dogs, values were 100% morbidity and 86% mortality (6/7). Brain impression smear slides made from all dogs that did not survive challenge exposure were CDV positive by use of a direct fluorescein isothiocyanate method. CONCLUSIONS The canarypox virus-CDV combination vaccine, administered SC or IM, is a safe product that elicits CDV seroconversion, does not interfere with other vaccine components, and protects vaccinated pups against virulent CDV challenge exposure.


title: [Cardiopulmonary exercise testing--its application in cardiology and occupational medicine].

text: Cardiopulmonary exercise testing is a method used to assess the exercise capacity. It is used in cardiology to define the diagnostic and prognostic information, the treatment and its effectiveness. This method is also useful in sport medicine and in occupational medicine. The cardiopulmonary exercise test involves measuring of gas exchange during exercise testing. The article presents the main parameters assessed during the test and the indications and contraindications for conducting the test. It also reveals the results of recently published clinical trials on the use of cardiopulmonary exercise test in patients with cardiovascular disease and in the working population. The study included variability of respiratory parameters during the cardiopulmonary exercise test and after its completion, as well as their impact on the prognostic value. In addition, the results of a study involving an optimal choice of interval training on the basis of oxygen consumption at peak exercise are summarized.


Using blended learning in training the public health workforce in emergency preparedness.


docid: gbzhdlnj

title: Absence of psychosis may influence linkage results for bipolar disorder.


title: Antioxidant and antimicrobial activities of native Rosa sp. from British Columbia, Canada.

text: Indigenous traditional knowledge and western science have revealed the potential for significant nutritional and therapeutic benefits among natural antioxidants. We investigated antioxidant and antimicrobial activity of rose hip extracts (Rosa nutkana, Rosa pisocarpa and Rosa woodsii) from wild British Columbia populations using liposome oxidation and disc diffusion assays. All extracts exhibited strong antioxidant activity. R. nutkana pericarp extracts contained high phenolic concentrations and showed greater antioxidant and antimicrobial activity than seed extracts. R. woodsii seed extracts had a higher phenolic concentration and greater antioxidant activity than pericarp extracts. Antioxidant activity was correlated with antimicrobial activity, and both extracts showed antimicrobial activity against yeast and Gram-positive bacteria. Our study is the first to demonstrate the antioxidant and antimicrobial properties of wild British Columbia roses. The effectiveness of selected species compared with standards demonstrates the significance of this natural resource to the continued health of human populations, and the need for conservation practices.


docid: mzghtnji

title: Early X-ray and CT appearances of severe acute respiratory syndrome: an analysis of 28 cases.

text: OBJECTIVE To study the early X-ray and CT findings of patients with severe acute respiratory syndrome (SARS). METHODS Chest radiography and CT were performed in 28 patients with SARS within one to three days after onset of the disease. CT examinations included conventional spiral CT and high-resolution CT (HRCT). The radiographic and CT findings of these patients were analyzed retrospectively. RESULTS Abnormal CT findings were noted in all the patients, but abnormal chest radiographic findings in 17 cases (60.7%, 17/28). CT showed single small focal patchy opacities in 23 patients (82.1%, 23/28), including oval ground-glass opacities in 20 patients, lobular distribution ground-glass opacities in 2 and small patchy consolidation in one. Multi-focal ground-glass opacities were found in 2 patients and extensive opacities in three. In the 28 patients, a total of 31 lesions were found in the upper (7, 22.6%), middle (3, 9.7%), and lower lobes (21, 67.7%). The diameter of the lesions ranged from 20 to 35 mm. CONCLUSION The dominant feature of early SARS patients is focal patchy opacity in the lung, and oval small ground-glass opacities are the common morphological findings on CT.


WHO inquiry into SARS leaves questions unanswered.