title: Murine coronavirus-induced hepatitis: JHM genetic background eliminates A59 spike-determined hepatotropism.

text: Recombinant murine coronaviruses, differing only in the spike gene and containing the strain A59 (moderately hepatotropic) and JHM (neurotropic) spike genes in the background of the JHM genome, were compared for the ability to replicate in the liver and induce hepatitis in weanling C57BL/6 mice. Interestingly, expression of the A59 spike glycoprotein within the background of the neurotropic JHM strain does not reproduce the A59 hepatotropic phenotype. Thus, the JHM genetic background plays a dominant role over the spike in the determination of hepatotropism.


title: Mouse studies of SARS coronavirus-specific immune responses to recombinant replication-defective adenovirus expressing SARS coronavirus N protein.

text: 1. A recombinant adenovirus encoding SARS coronavirus(SARS-CoV) nucleocapsid protein (rAd-N) was constructed. 2. The ability of the rAd-N to induce anti-SARS-CoV N antibody production and cellular immune responses was evaluated in an HLA-A2.1/Kb transgenic mouse model.


title: Roles of spike protein in the pathogenesis of SARS coronavirus.

text: 1. Infection with SARS coronavirus (SARS-CoV) induces a cellular stress condition known as the unfolded protein response (UPR). UPR induction is mediated primarily by viral spike (S) protein. The modulation of UPR by S protein involves activation of PERK protein kinase. Other branches of the UPR pathways controlled by IRE1 and ATF6 proteins, respectively, are not involved. 2. The protease inhibitor Ben-HCl effectively suppresses SARS-CoV infection by blocking virus entry. Viral infectivity is associated with the cleavage of S protein by the cellular protease factor Xa. 3. Two new aspects of the interaction between SARS-CoV S protein and the cell have been defined. These have important implications in the pathogenesis of SARS, providing opportunities for developing vaccines and antivirals against SARS-CoV. 4. Counteracting the UPR and targeting the cleavage of S protein with small molecule pharmaceutical agents represent two new anti-SARS-CoV strategies. 5. The receptor-binding domain of S protein delivered via adeno-associated virus can efficiently induce mucosal immunity and provide long-term protection against SARS-CoV infection.


title: MultiCode-PLx system for multiplexed detection of seventeen respiratory viruses.

text: The MultiCode-PLx system (EraGen Biosciences, Inc., Madison, WI) for the detection of respiratory viruses uses an expanded genetic alphabet, multiplex PCR chemistry, and microsphere flow cytometry to rapidly detect and specifically identify 17 different respiratory viruses directly in clinical specimens. The MultiCode-PLx system was tested in parallel with direct fluorescent-antibody (DFA) staining and rapid shell vial culture (R-mix cells; Diagnostic Hybrids, Inc. Athens, OH) with 354 respiratory specimens from adult patients that were submitted to the clinical virology laboratory at the Emory University Hospital. Single-target PCRs were performed with retained samples to confirm the positive results obtained with the MultiCode-PLx system for viruses not covered by DFA and R-mix culture (metapneumovirus, coronaviruses [CoV], parainfluenza viruses 4a and 4b, and rhinoviruses) and to resolve any discrepancies between the DFA and R-mix culture and the MultiCode-PLx results for viruses common to both systems. Respiratory viruses were detected in 77 (21.8%) and 116 (32.7%) specimens by DFA and R-mix culture and with the MultiCode-PLx system, respectively. Among the viruses common to both systems, the MultiCode-PLx system detected significantly more influenza A viruses (P = 0.0026). An additional increased diagnostic yield with the MultiCode-PLx system resulted from the detection of human metapneumovirus (HMPV) in 9 specimens, human CoV (HCoV) in 3 specimens, and human rhinovirus (HRV) in 16 specimens. Also, two mixed viral infections were detected by the MultiCode-PLx system (HCoV OC43 and HRV infections and HMPV and HRV infections), but none were detected by DFA and R-mix culture. Single-target PCRs verified the results obtained with the MultiCode-PLx system for 73 of 81 (90.1%) specimens that had discordant results or that were not covered by DFA and R-mix culture. The MultiCode-PLx system provides clinical laboratories with a practical, rapid, and sensitive means for the massively multiplexed molecular detection of common respiratory viruses.


title: 257 incidental appendectomies during total laparoscopic hysterectomy.

text: OBJECTIVE This retrospective observational report analyzes the demographics, blood loss, length of surgical duration, number of days in the hospital, and complications for 821 consecutive patients undergoing total laparoscopic hysterectomy over a 11-year period stratified by incidental appendectomy. METHODS A retrospective chart abstraction was performed. ANOVA and chi-square tests were performed with significance preset at P<0.05. RESULTS Of 821 consecutive patients undergoing total laparoscopic hysterectomy, 257 underwent elective appendectomy with the ultrasonic scalpel, either as part of their staging, treatment for pelvic pain, or prophylaxis against appendicitis. Comparing the 2 groups, no difference existed in mean age of 50+/-10 years or mean BMI of 27.6+/-6.7. Both groups had a similar mean blood loss of 130 mL. Surgery took less time (137 vs 118 minutes, P<0.0012) and the hospital stay was shorter in the appendectomy group (1.5 vs 1.2, P<0.0001) possibly because it was performed incidentally in most cases. No complications were attributable to the appendectomy, and complication types and rates in both groups were similar. Though all appendicies appeared normal, pathology was documented in 9%, including 3 carcinoid tumors. CONCLUSIONS Incidental appendectomy during total laparoscopic hysterectomy is not associated with significant risk and can be routinely offered to patients planning elective gynecologic laparoscopic procedures, as is standard for open procedures.


title: Comparison of transobturator tape surgery using commercial and hand made slings in women with stress urinary incontinence.

text: PURPOSE To compare the complications and success rates of hand-made sling with commercial sling used in transobturator tape (TOT) surgery. MATERIALS AND METHODS From 2008 to 2010, hand-made slings were used in TOT surgery, whereas commercial slings were used from 2010 to 2013 in our clinic. Overall 102 patients were included in the study. Patients were categorized into 2 groups: group 1 had hand-made (polypropylene monofilament) slings, while group 2 had commercial slings (polypropylene monofilament). We retrospectively reviewed 1-year follow-up results of the whole cohort. Ages, body mass indexes, menopausal status, operation time, cost of sling, success of operation and complications were recorded. All these data were compared between the 2 groups. RESULTS There were 41 patients (54.29 ± 9.88 years) in group 1 and 61 patients (52.82 ± 9.85 years) in group 2. Menopausal status and body mass index (28.1 vs. 29.2 kg/m² respectively) were similar for both groups. Previous history of incontinence or pelvic organ prolapse surgery (P = .046), mean duration of the procedure (P = .001), and vaginal extrusion rate (P = .016) were significantly lower in group 2. The cost of the sling was higher in group 2 than in group 1. There was no significant difference in success of operation between the groups (P = .319). CONCLUSION According to our results, hand-made mesh is a viable option in TOT surgery with similar efficacy, but surgeons should be careful in terms of vaginal extrusion.


What should change in nursing practice over the next five years?


title: Serum C-reactive protein in children with adenovirus infection.

text: OBJECTIVES (1) To evaluate serum C-reactive protein (CRP) concentrations in children with adenovirus infection, and (2) to compare CRP concentrations in adenovirus and influenza virus infection. PATIENTS AND METHODS Retrospective, comparative single-center study conducted in Emergency Department patients of a paediatric tertiary care center. Comparison of CRP in adenovirus infection and influenza was performed in patient groups stratified according to age and duration of fever. RESULTS In 87 children with adenovirus infection (median age, 1.5 years; interquartile range, 0.9-3.0), CRP levels of <2 mg/L, <10 mg/L, and <100 mg/L were found in 4 (4%), 12 (13%), and 66 (76%) patients, respectively. Median CRP in the children with adenovirus infection and in 130 children with influenza was 49 mg/L (21-96) and 9 mg/L (3-20), respectively (p = 0.001). A statistically significant difference remained when these 2 patient groups were stratified according to age (=2 vs. >2 years) and duration of fever (=3 vs. >3 days) (p <0.001). In adenovirus infection CRP concentrations were unrelated to age, duration of fever and severity of illness, as judged by the extent of mucosal involvement and by the frequency and duration of hospitalisation. CONCLUSION Paediatric adenovirus infection is associated with substantially elevated CRP concentrations in the absence of secondary bacterial infection. CRP levels were independent of the duration of illness, indicating that adenoviruses trigger an immediate inflammatory host response resembling invasive bacterial infection.


title: How long should we treat community-acquired pneumonia?

text: PURPOSE OF REVIEW The studies reviewed in this article suggest that a shorter duration of antibiotic therapy is comparable to standard therapy in the treatment of community-acquired pneumonia and promotes reduction of adverse events, microbial resistance, cost, and improved patient compliance. RECENT FINDINGS Community-acquired pneumonia has traditionally been treated with a 7-14-day course of antimicrobial therapy. Since there have been few well controlled trials regarding the optimal duration of therapy, however, there has been no consensus on length of therapy among different organizational guidelines. Several recent studies have demonstrated that shorter course antibiotic regimens are effective in the treatment of community-acquired pneumonia. SUMMARY Short-course antibiotic therapy is equivalent to standard length of therapy for clinical cure and bacterial eradication. Minimization of drug exposure, however, reduces selection pressure for resistant strains, strengthens patient compliance, and potentially reduces adverse events such as Clostridium difficile infections.


title: Imaging of unusual diffuse lung diseases.

text: PURPOSE OF REVIEW Computed tomography scans are commonly used in imaging lung diseases. As more information accumulates, patterns of rare or new diseases on CT scans are being increasingly reported. Several pulmonary diseases have distinguishing features, which are better delineated on high resolution CT scans than plain chest radiographs. The radiographic features of unusual diffuse lung diseases published in the past two years are described. RECENT FINDINGS Severe acute respiratory syndrome generally manifests as focal or diffuse bilateral areas of consolidation on chest radiography and reticulation with ground-glass attenuation commonly seen on CT scans. A normal HRCT rules out the diagnosis of Pneumocystis carinii pneumonia while a normal chest radiograph does not. Immunocompromised patients without AIDS, who have CMV pneumonia, generally demonstrate a combination of ground-glass attenuation, air-space consolidation, and small nodules on HRCT. Nodules less than 10 mm in size in immunocompromised patients are highly suggestive of viral infections. Bronchial wall thickening on HRCT associated with cavitating nodules is suggestive of Wegner granulomatosis in the appropriate clinical setting. Small cysts may be seen in a minority of patients with subacute hypersensitivity pneumonitis and centrilobular emphysema in chronic farmer's lung. Reversed halo sign has a high specificity for cryptogenic organizing pneumonia. The triad of ground-glass opacities, ill-defined centrilobular nodules and cysts and focal areas of air trapping is highly suggestive of subacute hypersensitivity pneumonitis. SUMMARY Familiarizing with radiographic and CT scan patterns may help the clinician to exclude certain diagnoses and narrow the differential diagnosis for others.


title: Circulating lupus anticoagulant and probable systemic lupus erythematosus in a cat.

text: An adult domestic short hair cat was presented in a critical condition with icterus. Various investigations demonstrated the presence of haemolytic anaemia and hepatic abnormalities, as well as significant coagulation defects. Systemic lupus erythematosus (SLE) was suggested as a possible cause.


Purpose: Brain metastases are a common complication of cancer and occur in about 15 - 40 % of patients with malignancies. The aim of this retrospective study was to differentiate between metastases from different primary tumors/CNS lymphyomas using morphologic criteria, fractional anisotropy (FA) and apparent diffusion coefficient (ADC). Materials and Methods: Morphologic criteria such as hemorrhage, cysts, pattern of contrast enhancement and location were reported in 200 consecutive patients with brain metastases/primary CNS lymphomas. FA and ADC values were measured in regions of interest (ROIs) placed in the contrast-enhancing tumor part, the necrosis and the non-enhancing peritumoral region (NEPTR). Differences between histopathological subtypes of metastases were analyzed using non-parametric tests, decision trees and hierarchical clustering analysis. Results: Significant differences were found in morphologic criteria such as hemorrhage or pattern of contrast enhancement. In diffusion measurements, significant differences between the different tumor entities were only found in ADC analyzed in the contrast-enhancing tumor part. Among single tumor entities, primary CNS lymphomas showed significantly lower median ADC values in the contrast-enhancing tumor part (ADClymphoma 0.92 [0.83 - 1.07] vs. ADCno_lymphoma 1.35 [1.10 - 1.64] P = 0.001). Further differentiation between types of metastases was not possible using FA and ADC. Conclusion: There were morphologic differences among the main subtypes of brain metastases/CNS lymphomas. However, due to a high variability of common types of metastases and low specificity, prospective differentiation remained challenging. DTI including FA and ADC was not a reliable tool for differentiation between different histopathological subtypes of brain metastases except for CNS lymphomas showing lower ADC values. Biopsy, surgery and staging remain essential for diagnosis. Key Points: • Histopathological subtypes of brain metastases/CNS lymphomas show different morphologic features on MRI• Primary CNS lymphomas show significantly reduced ADC values• DTI is not a reliable tool for differentiation between brain metastases Citation Format: • Bette S, Wiestler B, Delbridge C et al. Discrimination of Different Brain Metastases and Primary CNS Lymphomas Using Morphologic Criteria and Diffusion Tensor Imaging. Fortschr Röntgenstr 2016; 188: 1134 - 1143.


title: Over expression of a synthetic gene encoding interferon lambda using relative synonymous codon usage bias in Escherichia coli.

text: Interferon Lambda (IFN-λ) is a type III interferon which belongs to a novel family of cytokines and possesses antiviral and antitumor properties. It is unique in its own class of cytokines; because of the specificity towards its heterodimer receptors and its structural similarities with cytokines of other classes. This renders IFN-λ a better choice for the treatment against many diseases including viral hepatitis and human coronavirus (HCoV-EMC). The present study describes a computational approach known as relative synonymous codon usage (RSCU); used to enhance the expression of IFN-λ protein in a eukaryotic expression system. Manually designed and commercially synthesized IFN-λ gene was cloned into pET-22b expression plasmid under the control of inducible T7-lac promoter. Maximum levels of IFN-λ expression was observed with 0.4 mM IPTG in transformed E. coli incubated for 4 hours in LB medium. Higher concentrations of IPTG had no or negative effect on the expression of IFN-λ. This synthetically over expressed IFN-λ can be tested as a targeted treatment option for viral hepatitis after purification.


title: Pandemic flu: lessons from the Toronto SARS outbreak.

text: A review of the severe acute respiratory syndrome outbreak in Ontario in 2003 found that there had been several faults in healthcare services and that these had contributed to the spread of infection and the deaths of two nurses. This article argues that, six years on, these faults have not been rectified and that Ontario healthcare services may be unprepared for an H1N1 outbreak. The assistant deputy public health minister of Toronto offers her response.


title: Neuropathology of multiple sclerosis-new concepts.

text: Multiple sclerosis is a chronic inflammatory disease of the central nervous system with profound heterogeneity in clinical course, neuroradiological presentation and response to therapy. The pathological analysis of 235 actively demyelinating lesions coming from three centers revealed different structural and immunological features suggesting that different pathogenetic mechanisms are involved in lesion formation. On the basis of the presence or absence of immunoglobulin and complement deposition, myelin protein loss and the patterns of oligodendrocyte degeneration beside a T cell- and macrophage-dominated immune response, four distinct patterns of demyelination have been identified. In this short review, possible paraclinical markers for tissue destruction on the basis of the main features of myelin destruction are discussed. Furthermore, the importance of early axonal damage in multiple sclerosis is highlighted.


title: Identification of antibody against porcine coronavirus in human milk.

text: A total of 239 human milk samples collected from mothers in Chiba City, Japan during 1994-1997 were tested for the presence of anti-coronavirus antibody. Interestingly, twelve human milk samples were positive for IgA against porcine transmissible gastroenteritis coronavirus, and this represented 5%. These findings provided evidence that the interspecies transmission of coronavirus between human and porcine might occur in nature. This report is noteworthy because it is the first, to the best of our knowledge, demonstrating the presence of antibody against porcine transmissible gastroenteritis coronavirus in human milk.


title: Spontaneous pneumomediastinum in patients with severe acute respiratory syndrome.

text: Spontaneous pneumomediastinum (SP) unrelated to assisted ventilation is a newly recognised complication of severe acute respiratory syndrome (SARS). The objective of the present study was to examine the incidence, risk factors and the outcomes of SP in a cohort of SARS victims from a community outbreak. Data were retrieved from a prospectively collected database of virologically confirmed SARS patients. One hundred and twelve cases were analysable, with 13 patients developing SP (11.6%) at a mean +/- SD of 19.6 +/- 4.6 days from symptom onset. Peak lactate dehydrogenase level was associated with the development of SP. SP was associated with increased intubation and a trend towards death. Drainage was required in five cases. For patients who survived, the SP and/or the associated pneumothoraces took a median of 28 days (interquartile range: 15-45 days) to resolve completely. In conclusion, spontaneous pneumomediastinum appeared to be a frequent complication of severe acute respiratory syndrome. Further research is needed to investigate its pathogenesis.


title: [Viral Hepatitis].

text: HBV and HCV infection proceeds to chronic infection, and anti-cancer chemotherapy is contraindicated in patients with decompasated liver cirrhosis caused by both viruses. Reactivation of HBV refers to an increase in the HBV load caused by chemotherapy in a patient with HBV infection. Reactivation of HBV is classified into 2 categories: reactivation in a carrier and reactivation in a patient with resolved HBV infection. Because hepatitis caused by HBV reactivation is more likely to become severe and makes the treatment of original diseases difficult, it is most important to prevent the onset of hepatitis. The basic strategy for prevention and treatment of HBV reactivation associated with powerful chemotherapy regimens should follow the guidelines for the management of hepatitis B virus infection by the Japan Society of Hepatology. Risk of HBV reactivation is determined by HBV infection status and the degree of immunosuppression. HBV infection status is classified into chronic active hepatitis, inactive carrier, and resolved infection. This corresponds, in descending order, to the risk of reactivation. Patients undergoing chemotherapy should be screened for HBV infection. HBsAg levels should be measured in all patients prior to commencement of treatment. In HBsAg-positive patients, HBeAg, anti-HBe antibody, and HBV DNA levels should also be measured. In HBsAg-negative patients, anti-HBc antibody and anti-HBs antibody should be measured. The next step for patients with resolved HBV infection is measurement of HBV DNA levels. In patients with resolved HBV infection whose HBV DNA level is 20 IU/mL or higher, as in inactive carriers, prophylactic therapy should be commenced before chemotherapy. In patients with resolved HBV infection whose HBV DNA level is less than 20 IU/mL, periodic monitoring of HBV DNA should be performed during and after chemotherapy. Preemptive therapy should be started immediately if the HBV DNA level ex- ceeds 20 IU/mL.


title: Short internal sequences involved in replication and virion accumulation in a subviral RNA of turnip crinkle virus.

text: cis-acting sequences and structural elements in untranslated regions of viral genomes allow viral RNA-dependent RNA polymerases to correctly initiate and transcribe asymmetric levels of plus and minus strands during replication of plus-sense RNA viruses. Such elements include promoters, enhancers, and transcriptional repressors that may require interactions with distal RNA sequences for function. We previously determined that a non-sequence-specific hairpin (M1H) in the interior of a subviral RNA (satC) associated with Turnip crinkle virus is required for fitness and that its function might be to bridge flanking sequences (X. Sun and A. E. Simon, J. Virol. 77:7880-7889, 2003). To establish the importance of the flanking sequences in replication and satC-specific virion repression, segments on both sides of M1H were randomized and subjected to in vivo functional selection (in vivo SELEX). Analyses of winning (functional) sequences revealed three different conserved elements within the segments that could be specifically assigned roles in replication, virion repression, or both. One of these elements was also implicated in the molecular switch that releases the 3' end from its interaction with the repressor hairpin H5, which is possibly involved in controlling the level of minus-strand synthesis.


title: Epidemiologic evaluation of diarrhea in dogs in an animal shelter.

text: OBJECTIVES To determine associations among infectious pathogens and diarrheal disease in dogs in an animal shelter and demonstrate the use of geographic information systems (GISs) for tracking spatial distributions of diarrheal disease within shelters. SAMPLE POPULATION Feces from 120 dogs. PROCEDURE Fresh fecal specimens were screened for bacteria and bacterial toxins via bacteriologic culture and ELISA, parvovirus via ELISA, canine coronavirus via nested polymerase chain reaction assay, protozoal cysts and oocysts via a direct fluorescent antibody technique, and parasite ova and larvae via microscopic examination of direct wet mounts and zinc sulfate centrifugation flotation. RESULTS Salmonella enterica and Brachyspira spp were not common, whereas other pathogens such as canine coronavirus and Helicobacter spp were common among the dogs that were surveyed. Only intestinal parasites and Campylobacterjejuni infection were significant risk factors for diarrhea by univariate odds ratio analysis. Giardia lamblia was significantly underestimated by fecal flotation, compared with a direct fluorescent antibody technique. Spatial analysis of case specimens by use of GIS indicated that diarrhea was widespread throughout the entire shelter, and spatial statistical analysis revealed no evidence of spatial clustering of case specimens. CONCLUSIONS AND CLINICAL RELEVANCE This study provided an epidemiologic overview of diarrhea and interacting diarrhea-associated pathogens in a densely housed, highly predisposed shelter population of dogs. Several of the approaches used in this study, such as use of a spatial representation of case specimens and considering multiple etiologies simultaneously, were novel and illustrate an integrated approach to epidemiologic investigations in shelter populations.


title: Folding of the SARS coronavirus spike glycoprotein immunological fragment (SARS_S1b): thermodynamic and kinetic investigation correlating with three-dimensional structural modeling.

text: Spike glycoprotein of SARS coronavirus (S protein) plays a pivotal role in SARS coronavirus (SARS_CoV) infection. The immunological fragment of the S protein (Ala251-His641, SARS_S1b) is believed to be essential for SARS_CoV entering the host cell through S protein-ACE-2 interaction. We have quantitatively characterized the thermally induced and GuHCl-induced unfolding features of SARS_S1b using circular dichroism (CD), tryptophan fluorescence, and stopped-flow spectral techniques. For the thermally induced unfolding at pH 7.4, the apparent activation energy (E(app)) and transition midpoint temperature (Tm) were determined to be 16.3 +/- 0.2 kcal/mol and 52.5 +/- 0.4 degrees C, respectively. The CD spectra are not dependent on temperature, suggesting that the secondary structure of SARS_S1b has a relatively high thermal stability. GuHCl strongly affected SARS_S1b structure. Both the CD and fluorescent spectra resulted in consistent values of the transition middle concentration of the denaturant (Cm, ranging from 2.30 to 2.45 M) and the standard free energy change (deltaG(o), ranging from 2.1 to 2.5 kcal/mol) for the SARS_S1b unfolding reaction. Moreover, the kinetic features of the chemical unfolding and refolding of SARS_S1b were also characterized using a stopped-flow CD spectral technique. The obvious unfolding reaction rates and relaxation times were determined at various GuHCl concentrations, and the Cm value was obtained, which is very close to the data that resulted from CD and fluorescent spectral determinations. Secondary and three-dimensional structural predictions by homology modeling indicated that SARS_S1b folded as a globular-like structure by beta-sheets and loops; two of the four tryptophans are located on the protein surface, which is in agreement with the tryptophan fluorescence result. The three-dimensional model was also used to explain the recently published experimental results of S1-ACE-2 binding and immunizations.


title: Epidemiological survey of human metapneumovirus infection in a large pediatric tertiary care center.

text: The human metapneumovirus (hMPV) was recently identified and linked to acute respiratory tract infections (ARTI). To assess the clinical importance of this virus in infants and children, we developed a rapid and efficient reverse transcription-PCR-based screening method for a large volume of samples and tested retrospectively a collection of 1,132 respiratory specimens submitted over a full year period to the virology laboratory of a large tertiary care pediatric center in Montreal, Canada. A total of 41 samples from 37 patients were positive by this method. During the winter months of 2001, up to 8% of specimens submitted for respiratory virus testing were hMPV positive. Sequencing data of the hMPV M gene revealed that two genogroups of the virus, each of which can be divided into two subgroups, cocirculated during this time period. A case-controlled study was conducted to compare the symptoms associated with hMPV infection with those involving other etiologic agents causing ARTI. Symptoms most frequently observed in hMPV-positive patients were cough, wheezing, and dyspnea, although the symptomatology could differ substantially from patient to patient. No distinct symptom profile could be associated with hMPV. Three nosocomial cases of hMPV infection were identified. Together, our data suggest that hMPV is a significant cause of symptomatic respiratory tract infections in infants and children. The incidence of the disease and the morbidity associated with the infection justify adding hMPV to the list of common respiratory viruses routinely screened for by clinical laboratories.


title: Comparison of the Seeplex reverse transcription PCR assay with the R-mix viral culture and immunofluorescence techniques for detection of eight respiratory viruses.

text: This study evaluated the clinical usefulness of a newly introduced multiplex reverse transcription PCR assay (Seeplex RV; Seegene, Inc., Seoul, Korea) in patients with respiratory symptoms. Fifty clinical respiratory specimens (45 from children, 5 from adults) were tested for 8 viruses (influenza virus type A and B, parainfluenza virus type 1, 2, 3, respiratory syncytial virus type A and B, and adenovirus) by Seeplex RV (S-RV) and R-mix viral culture with immunofluorescence (VC-IF). Forty (80%) of the 50 samples showed concordant results between S-RV and VC-IF; 24 of these showed the same positive and 16 showed the same negative results. Among the 10 discrepant samples, 9 were S-RV-positive and VC-IF-negative. Six were obtained in patients with lower respiratory tract infection. Only 1 sample was VC-IF-positive and S-RV-negative. This patient had pneumonia. In 3 cases, more than 1 virus was identified by S-RV. The total running time of S-RV was 6 hr, which shortens the detection time for the viral presence by 2 workdays compared to VC-IF. In conclusion, S-RV is reliable, rapid, relatively easy to perform, and able to detect more than 1 virus simultaneously. Therefore, implementation of the S-RV assay in clinical laboratories will aid rapid diagnosis and treatment of major viral infections of the respiratory tract.


title: Viral pathogens of acute lower respiratory tract infection in China.

text: OBJECTIVES To document the viral etiology of acute lower respiratory tract infection (ALRIs) in Chinese children. SETTING Children Hospital, Zhejiang University, China. STUDY DESIGN Cross-sectional. PARTICIPANTS 34885 children with ALRI between January 2001 to December 2006. METHODS Nasopharyngeal aspirates were collected from all subjects. Respiratory syncytial virus (RSV), adenovirus (ADV), type 1 to 3 parainfluenza viruses (PIV), and type A and B influenza virus (Flu) were detected by direct immunofluorescence. RESULTS Viruses were identified in 32.3% cases, including RSV (23.6%), PIV 3 (4.3%), Flu A (2.0%), ADV (1.7%), PIV I (0.6%), Flu B (0.2%) and PIV 2 (0.1%). RSV and PIV 3 predominated in younger children while Flu A and Flu B predominated in older children (P<0.001, respectively). PIV 1 was more prevalent in children aged 1 to 3 years. The peak frequency of RSV, PIV 3 and Flu A were in early spring, June to August, and August and September, respectively. Flu B had a peak in the winter and spring. Adenovirus infections occurred in all seasons with a relatively constant frequency. CONCLUSIONS Viruses are an important cause of ALRIs in Chinese children constituting 1/3 of total cases. RSV is the most common pathogen.


title: Contamination of transplantable tumors, cell lines, and monoclonal antibodies with rodent viruses.

text: Different biological materials were tested for murine viral contamination by using the mouse/rat antibody production test. Of 297 tumors examined, 75 (25.3%) were contaminated. Considerable differences in the contamination rate became evident when transplantable tumors from in vitro and from in vivo passages were compared. Of 186 tumors that had been propagated in animals, 36.6% were positive, whereas only 7 of 111 (6.3%) tumors propagated in vitro were contaminated. The highest rate of contamination was detected in mouse tumors. Testing of 135 specimens of mouse origin revealed 46.7% were contaminated, and 57 (70.4%) of 81 samples propagated in mice were positive for murine viruses. Moreover, 6.7% of 90 human tumors that had been passaged in athymic nude mice and 3.5% of 57 rat tumors were positive. Lymphocytic choriomeningitis virus was detected in 4 of 14 hamster tumors. The most frequent contaminant was lactic dehydrogenase elevating virus followed by reovirus 3, lymphocytic choriomeningitis virus, minute virus of mice, mouse hepatitis virus, rat coronaviruses, Kilham rat virus, and Mycoplasma pulmonis. Contamination with reovirus 3 and minute virus of mice was found in 4 (3.7%) of 109 cell lines tested, and 2 of 60 monoclonal antibody preparations or hybridoma cells contained lactic dehydrogenase virus. Contamination with two pathogens was detected in four mouse tumors and in one cell line.


Governments in a dilemma over bird flu.


Characterization of [3H]adenosine binding to fat cell membranes.


Murine coronavirus RNA.


title: Perspectives on the basic reproductive ratio.

text: The basic reproductive ratio, R0, is defined as the expected number of secondary infections arising from a single individual during his or her entire infectious period, in a population of susceptibles. This concept is fundamental to the study of epidemiology and within-host pathogen dynamics. Most importantly, R0 often serves as a threshold parameter that predicts whether an infection will spread. Related parameters which share this threshold behaviour, however, may or may not give the true value of R0. In this paper we give a brief overview of common methods of formulating R0 and surrogate threshold parameters from deterministic, non-structured models. We also review common means of estimating R0 from epidemiological data. Finally, we survey the recent use of R0 in assessing emerging diseases, such as severe acute respiratory syndrome and avian influenza, a number of recent livestock diseases, and vector-borne diseases malaria, dengue and West Nile virus.


title: Subsensitivity to beta-adrenergic stimulation in atria from rats infested with Syphacia sp.

text: Syphacia sp. is a common intestinal parasite in conventionally-housed laboratory rodents. Although gross lesions are rare in oxyuriasis, it is possible that more subtle changes may develop, which may affect research results. In this study, we analysed the responsiveness to beta-adrenergic stimulation by isoproterenol (ISO) of left atria isolated from Syphacia-infested (SYPH) and control, non-infested adult male Wistar rats (CONT). In the non-infested animals, ISO pD(2) was not significantly changed by ivermectin treatment. Whereas the maximal inotropic response to ISO was not significantly affected, the pD(2) value was decreased in SYPH (7.61 +/- 0.09, n = 7, vs 8.21 +/- 0.25 in CONT, n = 5, P < 0.05), indicating lower sensitivity to beta-adrenergic stimulation. This change was similar to that caused by a classic stressor, namely repeated immobilization, in non-infested rats (IMMO). In this group, ISO pD(2) was 7.62 +/- 0.14, n = 6 (P < 0.05 with relation to CONT). The results indicate that infestation with Syphacia sp. is as effective as immobilization at diminishing cardiac reactivity to beta-adrenergic stimulation. It is thus possible that oxyuriasis may affect the response of other tissues to physiological modulators.


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title: Diagnosis of severe acute respiratory syndrome (SARS) by detection of SARS coronavirus nucleocapsid antibodies in an antigen-capturing enzyme-linked immunosorbent assay.

text: Recombinant severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein was employed to establish an antigen-capturing enzyme-linked immunosorbent assay (ELISA). Antinucleocapsid protein antibodies could be detected in 68.4% of probable SARS patients 6 to 10 days after illness and in 89.6% of the patients 11 to 61 days after illness. No false-positive results were observed in 20 non-SARS fever patients, 24 non-SARS respiratory illness patients, and 20 health care workers. Among 940 other non-SARS clinical serum samples, only 1 was found to be weakly positive. This method provides a new, sensitive, and specific approach for SARS diagnosis.


title: Therapeutic antibodies.

text: Monoclonal antibodies had the lure of drugs very much since their first description. The ability to bind to a predetermined chemical structure stimulated the imagination of drug discoverers and developers. Nevertheless it took many years before a drug was registered which started to make good on the promise. The complexity of the molecule, made up of four polypeptide chains, its large molecular weight, its multiple and versatile functional domains and its mouse origin initially were obstacles for the production and the utilisation. Also the selection of appropriate target structures on the surface of cells turned out be difficult. Many of these difficulties have been overcome. The replacement of most of the murine sequences with equivalent human sequences and the concomittant decrease in immunogenicity, and the identification of cell surface components which are causative and limiting in cellular transformation have made monoclonal antibodies valuable weapons in the fight against cancer. Multiple mechanisms of monoclonal antibody action are being exploited for this purpose. Antibodies can sequester growth factors and prevent the activation of crucial growth factor receptors. A monoclonal antibody directed against the vascular endothelial growth factor (VEGF) has been shown to be a potent neo-vascularisation inhibitor (bevacizumab). An antibody against the extracellular domain of the EGF receptor prevents the binding of the ligand to the receptor and thereby its activation (cetuximab). EGFR activity, however, is absolutely required for the survival and proliferation of certain human tumour cells. An antibody which interferes with the dimerisation of the ErbB2 and the ErbB3 members of the EGF receptor family prevents the association of a most potent signaling module (pertuxumab). The signals emenating from this dimer determine many phenotypic properties of e.g. human breast cancer cells. A monoclonal antibody also directed against ErbB2 has been most successful, clinically and commercially (trastuzumab). This antibody interferes with signals generated by the receptor and causes the arrest of the cell cycle in tumour cells. In addition, it recruits immune effector cells as cytotoxic agents. Finally, monoclonal antibody derivatives, single chain Fv fragments, have been used as a basis for the construction of recombinant tumour toxins. These molecules harness the exquisite binding specificity of the antibodies and combine them with the toxic principles of bacteria.


title: Human papillomavirus type 16 E7 peptide-directed CD8+ T cells from patients with cervical cancer are cross-reactive with the coronavirus NS2 protein.

text: Human papillomavirus type 16 (HPV16) E6 and E7 oncoproteins are required for cellular transformation and represent candidate targets for HPV-specific and major histocompatibility complex class I-restricted CD8(+)-T-cell responses in patients with cervical cancer. Recent evidence suggests that cross-reactivity represents the inherent nature of the T-cell repertoire. We identified HLA-A2 binding HPV16 E7 variant peptides from human, bacterial, or viral origin which are able to drive CD8(+)-T-cell responses directed against wild-type HPV16 E7 amino acid 11 to 19/20 (E7(11-19/20)) epitope YMLDLQPET(T) in vitro. CD8(+) T cells reacting to the HLA-A2-presented peptide from HPV16 E7(11-19(20)) recognized also the HLA-A2 binding peptide TMLDIQPED (amino acids 52 to 60) from the human coronavirus OC43 NS2 gene product. Establishment of coronavirus NS2-specific, HLA-A2-restricted CD8(+)-T-cell clones and ex vivo analysis of HPV16 E7 specific T cells obtained by HLA-A2 tetramer-guided sorting from PBL or tumor-infiltrating lymphocytes obtained from patients with cervical cancer showed that cross-reactivity with HPV16 E7(11-19(20)) and coronavirus NS2(52-60) represents a common feature of this antiviral immune response defined by cytokine production. Zero of 10 patients with carcinoma in situ neoplasia and 3 of 18 patients with cervical cancer showed > or =0.1% HPV16 E7-reactive T cells in CD8(+) peripheral blood lymphocytes. In vivo priming with HPV16 was confirmed in patients with cervical cancer or preinvasive HPV16-positive lesions using HLA-A2 tetramer complexes loaded with the E6-derived epitope KLPQLCTEL. In contrast, we could not detect E6-reactive T cells in healthy individuals. These data imply that the measurement of the HPV16 E7(11-19(20)) CD8(+)-T-cell response may reflect cross-reactivity with a common pathogen and that variant peptides may be employed to drive an effective cellular immune response against HPV.


title: A serologic survey for viruses and Mycoplasma pulmonis among wild house mice (Mus domesticus) in southeastern Australia.

text: Plasma samples from 267 wild house mice (Mus domesticus) trapped at 14 sites in southeastern Australia were screened for antibody to 14 viruses normally associated with laboratory-reared rodents and to Mycoplasma pulmonis. Serologic prevalence was high for murine cytomegalovirus (99%, n = 94), murine coronavirus (95%), and murine rotavirus (74%). Samples from mice collected at all sites contained antibody to these viruses. The serologic prevalence was lower for mouse adenovirus, strain K87 (37%), parvovirus (33%), and reovirus type 3 (28%), with substantial site-to-site variation. Plasma from mice collected at 12 sites contained mouse adenovirus or reovirus antibody, and samples from mice at eight sites contained parvovirus antibody. Parvovirus-antibody positive mice were typically from high density populations or from low density populations that had recently declined from high density. Antibody to lymphocytic choriomeningitis virus (LCMV) and Sendai virus occurred at only three sites, and the serologic prevalence was very low (9.6% and 1.8%, respectively). All of the LCMV-positive mice were from northeastern New South Wales. The presence of this zoonotic virus in a mouse plague-prone region raises questions about human health risks resulting from cohabitation with large numbers of mice. It appeared that mouse populations at high density or declining from high density had higher prevalence of viral antibody than populations that had been at low or moderate density for some time. Thus, viral epizootics may occur among high-density populations and may be responsible for or precipitate declines in mouse density. These data raise the possibility of rodent viruses having potential as biological control agents.


title: [Initial radiographic manifestations in patients with SARS].

text: OBJECTIVE To observe initial radiographic manifestations and evaluate its value for the diagnosis and differential diagnosis in patients with severe acute respiratory syndrome (SARS). METHODS Initial chest radiographs of 79 patients, 15 to 69 years old (mean age 34.1 years, 40 men and 39 women), were retrospectively reviewed in the study. For all patients, average temperature was 38.8 degrees C and mean time from onset of the fever to the time of initial radiographs taken was 5.8 days. Location, abnormal distribution, appearance of lung abnormalities, effusion of pericadiaum and pleura, and lymphadenopathy were observed. Abnormal distribution and associated findings were compared. Differences of mean age, temperature, and the time were statistically analyzed between patients with and without abnormal findings. RESULTS Initial chest radiographs were normal in 36 of 79 patients (45.5%). Abnormal radiographs in 43 patients (81.3%) showed exudative changes. No significant difference of mean age and day (form onset of the fever to the time of initial radiographs taken) was found between the patients with and without abnormal initial chest radiographs (P > 0.05). No lymphadenopathy, effusion of pericadiaum and pleura were found on the initial chest radiographs. CONCLUSIONS The main abnormal radiographic appearance of lung in patients with SARS is exudative changes. SARS patients with normal initial chest radiographs are not uncommon and abnormal radiographic findings may appear to be late after onset of fever. Therefore, more attention should be paid in the clinical diagnosis of SARS.


title: Organization of two transmissible gastroenteritis coronavirus membrane protein topologies within the virion and core.

text: The difference in membrane (M) protein compositions between the transmissible gastroenteritis coronavirus (TGEV) virion and the core has been studied. The TGEV M protein adopts two topologies in the virus envelope, a Nexo-Cendo topology (with the amino terminus exposed to the virus surface and the carboxy terminus inside the virus particle) and a Nexo-Cexo topology (with both the amino and carboxy termini exposed to the virion surface). The existence of a population of M molecules adopting a Nexo-Cexo topology in the virion envelope was demonstrated by (i) immunopurification of (35)S-labeled TGEV virions using monoclonal antibodies (MAbs) specific for the M protein carboxy terminus (this immunopurification was inhibited only by deletion mutant M proteins that maintained an intact carboxy terminus), (ii) direct binding of M-specific MAbs to the virus surface, and (iii) mass spectrometry analysis of peptides released from trypsin-treated virions. Two-thirds of the total number of M protein molecules found in the virion were associated with the cores, and one-third was lost during core purification. MAbs specific for the M protein carboxy terminus were bound to native virions through the M protein in a Nexo-Cexo conformation, and these molecules were removed when the virus envelope was disrupted with NP-40 during virus core purification. All of the M protein was susceptible to N-glycosidase F treatment of the native virions, which indicates that all the M protein molecules are exposed to the virus surface. Cores purified from glycosidase-treated virions included M protein molecules that completely or partially lost the carbohydrate moiety, which strongly suggests that the M protein found in the cores was also exposed in the virus envelope and was not present exclusively in the virus interior. A TGEV virion structure integrating all the data is proposed. According to this working model, the TGEV virion consists of an internal core, made of the nucleocapsid and the carboxy terminus of the M protein, and the envelope, containing the spike (S) protein, the envelope (E) protein, and the M protein in two conformations. The two-thirds of the molecules that are in a Nexo-Cendo conformation (with their carboxy termini embedded within the virus core) interact with the internal core, and the remaining third of the molecules, whose carboxy termini are in a Nexo-Cexo conformation, are lost during virus core purification.


title: Modelling the utility of body temperature readings from primary care consults for SARS surveillance in an army medical centre.

text: INTRODUCTION There is interest in surveillance systems for outbreak detection at stages where clinical presentation would still be undifferentiated. Such systems focus on detecting clusters of syndromes in excess of baseline levels, which may indicate an outbreak. We model the detection limits of a potential system based on primary care consults for the detection of an outbreak of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS Data from an averaged-sized medical centre were extracted from the Patient Care Enhancement System (PACES) [the electronic medical records system serving the Singapore Armed Forces (SAF)]. Thresholds were set to 3 or more cases presenting with particular syndromes and a temperature reading of >or=38oC (T >or=38). Monte Carlo simulation was used to insert simulated SARS outbreaks of various sizes onto the background incidence of febrile cases, accounting for distribution of SARS incubation period, delay from onset to first consult, and likelihood of presenting with T >or=38 to the SAF medical centre. RESULTS Valid temperature data was available for 2,012 out of 2,305 eligible syndromic consults (87.2%). T >or=38 was observed in 166 consults (8.3%). Simulated outbreaks would peak 7 days after exposure, but, on average, signals at their peak would consist of 10.9% of entire outbreak size. Under baseline assumptions, the system has a higher than 90% chance of detecting an outbreak only with 20 or more cases. CONCLUSIONS Surveillance based on clusters of cases with T >or=38 helps reduce background noise in primary care data, but the major limitation of such systems is that they are still only able to confidently detect large outbreaks.


title: Collaboration of antibody and inflammation in clearance of rabies virus from the central nervous system.

text: To investigate the involvement of various cellular and humoral aspects of immunity in the clearance of rabies virus from the central nervous system, (CNS), we studied the development of clinical signs and virus clearance from the CNS in knockout mice lacking either B and T cells, CD8+ cytotoxic T cells, B cells, alpha/beta interferon (IFN-alpha/beta) receptors, IFN-gamma receptors, or complement components C3 and C4. Following intranasal infection with the attenuated rabies virus CVS-F3, normal adult mice of different genetic backgrounds developed a transient disease characterized by loss of body weight and appetite depression which peaked at 13 days postinfection (p.i.). While these animals had completely recovered by day 21 p.i., mice lacking either B and T cells or B cells alone developed a progressive disease and succumbed to infection. Mice lacking either CD8+ T cells, IFN receptors, or complement components C3 and C4 showed no significant differences in the development of clinical signs by comparison with intact counterparts having the same genetic background. However, while infectious virus and viral RNA could be detected in normal control mice only until day 8 p.i., in all of the gene knockout mice studied except those lacking C3 and C4, virus infection persisted through day 21 p.i. Analysis of rabies virus-specific antibody production together with histological assessment of brain inflammation in infected animals revealed that clearance of CVS-F3 by 21 days p.i. correlated with both a strong inflammatory response in the CNS early in the infection (day 8 p.i.), and the rapid (day 10 p.i.) production of significant levels of virus-neutralizing antibody (VNA). These studies confirm that rabies VNA is an absolute requirement for clearance of an established rabies virus infection. However, for the latter to occur in a timely fashion, collaboration between VNA and inflammatory mechanisms is necessary.


title: Assessing the economic burden of avian infectious bronchitis on poultry farms in Brazil.

text: Avian infectious bronchitis (IB), caused by avian infectious bronchitis virus (IBV), is a worldwide endemic disease of chickens that affects all branches of the poultry industry. Multiple geno/serotypes occur, and lowvaccine cross-protection results from the highly divergent IBV types. In view of the lack of consistent data on the economic losses caused by IB and the poor protection resulting from the use of the Massachusetts type as a live vaccine in Brazil, this survey aimed to estimate the losses per 1000 birds in broiler and breeder flocks positive for IBV. Thirty-two different IBV genetic types were found. In breeders, the total loss per 1,000 birds was US $3567.4 and US $4210.8 at 25-26 and 42 weeks old, respectively, whereas in broilers (48 days old), the estimated loss was US $266.3 per 1,000 birds. Taken together, the results show a significant and measurable economic impact on the broiler and breeder industries, with an age-dependent increasing trend and an association with multiple genetic types of the virus.


title: A norovirus protease structure provides insights into active and substrate binding site integrity.

text: Norovirus 3C-like proteases are crucial to proteolytic processing of norovirus polyproteins. We determined the crystal structure of the 3C-like protease from Chiba virus, a norovirus, at 2.8-A resolution. An active site including Cys139 and His30 is present, as is a hydrogen bond network that stabilizes the active site conformation. In the oxyanion hole backbone, a structural difference was observed probably upon substrate binding. A peptide substrate/enzyme model shows that several interactions between the two components are critical for substrate binding and that the S1 and S2 sites appropriately accommodate the substrate P1 and P2 residues, respectively. Knowledge of the structure and a previous mutagenesis study allow us to correlate proteolysis and structure.


title: Totally laparoscopic resection and extraction of specimens via transanal route in synchronous colon and gastric cancer.

text: INTRODUCTION Synchronous colon and gastric cancer is a rare clinical entity. In the present case, it is aimed to show that in a patient with synchronous colon and gastric cancer, laparoscopic resection can be safely performed and both specimens can be delivered through a natural orifice. In our knowledge, this is the first example showing the delivery of the gastric resection specimen through the anus in a human being. CASE REPORT Sixty-six years old male patient with an upper gastrointestinal bleeding and obstruction symptoms was admitted to our department and the evaluation revealed an advanced stage gastric and a synchronous colon cancer. A laparoscopic palliative subtotal gastrectomy with a subtotal colectomy was performed. All anastomoses were performed intracorporeally and colectomy and subtotal gastrectomy specimens were successfully delivered via trans-anal route without any difficulty. Despite major abdominal organ resections, the patients required quite less analgesics in the postoperative period. DISCUSSION Specimen extraction through the natural orifices eliminates the need and problems of performing additional abdominal incisions to the patients which also leads to reduced postoperative pain. Synchronous surgical procedures do not prevent the natural orifice surgery.


title: Efficacy of Human Fibrinogen-Thrombin Patch (TachoSil) Clinical Application in Upper Gastrointestinal Cancer Surgery.

text: BACKGROUND Previous studies investigated the efficacy and applicability of tissue adhesives in gastrointestinal surgery while no evidence is available to date about a novel compound, TachoSil (Takeda, Zurich, Switzerland). The primary aim of this observational study was to assess the effect of new fibrin sealant on the incidence of postoperative complications in a homogeneous group of patients submitted to upper gastrointestinal surgery for cancer. METHODS Two cohorts of 28 and 34 patients undergoing upper gastrointestinal for surgery were compared. In the first cohort, the anastomotic site was treated with TachoSil fibrinogen-thrombin-collagen patches and in the second no collagen sponge or any other hemostatic sealant was used. Postoperative complications and outcomes as well as postoperative biochemical parameters were analyzed. RESULTS Postoperative complications occurred in 12 patients (35.3%) and 2 patients (7.1%) in control and collagen sponge group respectively (χ2 = 3.539, p < 0.05), with no anastomotic leakage in the collagen sponge group. A binary logistic regression analysis showed that the nonuse of collagen sponge [odds ratio (OR) = 0.025, 95% confidence interval (CI) = 0.001-0.457, p = 0.01] was independently associated with postoperative complication occurrence. CONCLUSIONS The addition of fibrinogen-thrombin-collagen sponge patch may reduce postoperative complication rate after upper gastrointestinal surgery for cancer. Further study to delineate the role of TachoSil in gastrointestinal surgery are also needed to demonstrate improved effectiveness and applicability.


title: Isolation of bovine coronavirus from feces and nasal swabs of calves with diarrhea.

text: Fecal and nasal samples were collected from 180 calves with diarrhea and 36 clinically normal co-habitants, and tested for virus using HRT-18 cell cultures derived from human rectal adenocarcinoma. A cytopathic virus was isolated from 5 fecal and 56 nasal samples obtained from diarrheic calves. All calves in which the virus was isolated from diarrheic feces were positive for virus isolation from nasal swabs. The virus was also isolated from the nasal swabs of 10 clinically normal calves that were co-habitants with diarrheic calves. Because they were morphologically similar to coronavirus, agglutinated mouse erythrocytes and serologically identical with the Nebraska calf diarrhea coronavirus, new isolates were identified as bovine coronavirus. The demonstration of viral antigens in nasal epithelial cells by a direct immunofluorescence was in close agreement with the virus isolation in HRT-18 cell cultures. This is the first report on the isolation of bovine coronavirus from newborn calves with diarrhea in Japan. The evidence that the virus was frequently isolated from nasal swabs is of great interest for understanding the pathogenesis of bovine coronavirus infection.


title: Isolation of Helicobacter canis from a colony of bengal cats with endemic diarrhea.

text: On the basis of biochemical, phenotypic, and 16S rRNA analyses, Helicobacter canis was isolated from Bengal cats with and without chronic diarrhea. Because the cats were coinfected with other potential pathogens, including Campylobacter helveticus, and because H. canis was isolated from nondiarrheic cats, the causal role of H. canis in producing the diarrhea could not be proven. Histologically, the colons of the four affected cats were characterized by mild to moderate neutrophilic, plasmacytic, and histocytic infiltrates in the lamina propria. Rare crypt abscesses were also noted for three cats but were a more prominent feature of the colonic lesions noted for the fourth cat. This is the first observation of H. canis in cats and raises the possibility that H. canis, like H. hepaticus and H. bilis in mice, can cause inflammation of the colon, particularly in hosts with immune dysregulation. Further studies are needed to determine the importance of H. canis as a primary enteric pathogen in cats and the role of cats in the possible zoonotic spread of H. canis to humans.


title: Mechanisms and enzymes involved in SARS coronavirus genome expression.

text: A novel coronavirus is the causative agent of the current epidemic of severe acute respiratory syndrome (SARS). Coronaviruses are exceptionally large RNA viruses and employ complex regulatory mechanisms to express their genomes. Here, we determined the sequence of SARS coronavirus (SARS-CoV), isolate Frankfurt 1, and characterized key RNA elements and protein functions involved in viral genome expression. Important regulatory mechanisms, such as the (discontinuous) synthesis of eight subgenomic mRNAs, ribosomal frameshifting and post-translational proteolytic processing, were addressed. Activities of three SARS coronavirus enzymes, the helicase and two cysteine proteinases, which are known to be critically involved in replication, transcription and/or post-translational polyprotein processing, were characterized. The availability of recombinant forms of key replicative enzymes of SARS coronavirus should pave the way for high-throughput screening approaches to identify candidate inhibitors in compound libraries.


title: Activation of the immune coagulation system by murine hepatitis virus strain 3.

text: Viral infections result in alterations in hemostasis and coagulation. It has previously been shown that susceptibility to murine hepatitis virus strain 3 (MHV-3), a coronavirus, correlates directly with the spontaneous, T lymphocyte-instructed expression of a procoagulant monokine that exhibits prothrombin-cleaving activity (procoagulant activity [PCA]). A biologic role for PCA in the pathogenesis of MHV-3 infection is suggested by results of in vivo microscopic observations made during acute MHV-3 infection. Recently, it has been demonstrated that prostaglandin E2 abrogates the induction of PCA by MHV-3 both in vivo and in vitro and prevents hepatic necrosis and the associated microcirculatory changes. These data suggest that MHV-3 induces cellular injury through the activation of the coagulation cascade and provide further evidence for a role for PCA in the pathogenesis of MHV-3 infection.


Inactivated influenza vaccine is safe for children and adults with stable asthma.


title: Two coronaviruses isolated from central nervous system tissue of two multiple sclerosis patients.

text: Two coronaviruses were isolated from brain material obtained at autopsy from two multiple sclerosis patients. The viruses were neutralized by serum and spinal fluid from these patients. Although most of the population have antibody to these virus isolates, multiple sclerosis patients have slightly higher concentrations of serum antibody than controls. The results suggest that coronaviruses should be considered as one additional virus with a potential implication in the etiology of multiple sclerosis.


title: Virologic and serologic identification of minute virus of canines (canine parvovirus type 1) from dogs in Japan.

text: Minute virus of canines (MVC), also known as canine parvovirus type 1, was initially believed to be a nonpathogenic agent, since it was first isolated from canine fecal specimens in the late 1960s. However, subsequent pathological as well as epidemiological studies suggested that MVC is a pathogen of neonatal puppies and is widely distributed among domestic dogs in the United States. The virus also has been shown to cause fetal deaths. Nevertheless, the virus was not detected in dogs outside the United States until recently, presumably because of a lack of widespread availability of the only susceptible canine cell line, WRCC/3873D, used for MVC isolation. We examined 470 clinical specimens from 346 dogs by PCR and detected MVC-specific gene fragments from four diseased puppies (positive rate, 1.2%). Viruses were recovered from three PCR-positive rectal specimens by using WRCC/3873D and MDCK cells. The isolates possessed antigenic and genomic properties similar to those of the U.S. reference strain GA3 and were identified as MVC. In addition, seroepidemiological evidence that 5.0% of dogs possessed anti-MVC antibodies also indicated the presence of MVC infection among dogs in Japan. From this study and several recent European reports describing MVC field cases, it is evident that MVC is distributed among domestic dogs worldwide.


title: Network Medicine Framework for Identifying Drug Repurposing Opportunities for COVID-19.

text: The COVID-19 pandemic demands the rapid identification of drug-repurpusing candidates. In the past decade, network medicine had developed a framework consisting of a series of quantitative approaches and predictive tools to study host-pathogen interactions, unveil the molecular mechanisms of the infection, identify comorbidities as well as rapidly detect drug repurpusing candidates. Here, we adapt the network-based toolset to COVID-19, recovering the primary pulmonary manifestations of the virus in the lung as well as observed comorbidities associated with cardiovascular diseases. We predict that the virus can manifest itself in other tissues, such as the reproductive system, and brain regions, moreover we predict neurological comorbidities. We build on these findings to deploy three network-based drug repurposing strategies, relying on network proximity, diffusion, and AI-based metrics, allowing to rank all approved drugs based on their likely efficacy for COVID-19 patients, aggregate all predictions, and, thereby to arrive at 81 promising repurposing candidates. We validate the accuracy of our predictions using drugs currently in clinical trials, and an expression-based validation of selected candidates suggests that these drugs, with known toxicities and side effects, could be moved to clinical trials rapidly.


title: Correlation of susceptibility of immature mice to fungal infection (blastomycosis) and effector cell function.

text: Immature mice are highly susceptible to blastomycosis, which is similar to other mycoses and has parallels in humans. The murine susceptibility is noteworthy in that it persists beyond the development of resistance to other, nonfungal pathogens and the maturation of most immune functions. As the susceptibility to blastomycosis appeared to be related to an early event after infection, primary effector cell function was studied. We found that peritoneal inflammatory cells, enriched for neutrophils, from immature (3-week-old) mice killed nonphagocytizable Blastomyces dermatitidis cells less (25%) than did cells from mature (8-week) mice (70%) (P<0.01), a defect intrinsic to the neutrophils. This correlated with an impaired immature cell oxidative burst. Killing of phagocytizable Candida albicans was not significantly different, 73 versus 87%. Thioglycolate-elicited cells were more impaired; killing of B. dermatitidis was insignificant, and killing of C. albicans was more impaired in immature (16% killing) than in mature (45%) cells (P<0.02). Peripheral blood neutrophils from mature animals killed B. dermatitidis (41%) more than did those from immature animals (10%) (P<0.02); C. albicans was killed efficiently by both. Resting or activated peritoneal macrophages from both types of animals showed no differences in B. dermatitidis killing. These results suggest that the susceptibility of immature mice is related at least in part to the depressed capacity of their neutrophils to kill B. dermatitidis.


title: Advances in the value of eggs and egg components for human health.

text: The avian egg is an important source of nutrients, containing all of the proteins, lipids, vitamins, minerals, and growth factors required by the developing embryo, as well as a number of defense factors to protect against bacterial and viral infection. Moreover, eggs are now understood to contain substances with biological functions beyond basic nutrition, and extensive research has been undertaken to identify and characterize these biologically active components. This review mainly focused on biological activities of proteins and peptides derived from egg components. Several biological activities have now been associated with egg components, including novel antimicrobial activities, antiadhesive properties, immunomodulatory, anticancer, and antihypertensive activities, antioxidant properties, protease inhibitors, nutrient bioavailability, and functional lipids, highlighting the importance of egg and egg components in human health and in disease prevention and treatment. Continued research to identify new and existing biological functions of hen egg components will help to define new methods to further improve the value of eggs as a source of numerous biologically active compounds with specific benefits for human and animal health and secure their role in the therapy and prevention of chronic and infectious disease.


title: Receptor-binding domain of SARS-Cov spike protein: soluble expression in E. coli, purification and functional characterization.

text: AIM Spike protein of coronavirus is responsible for virus binding, fusion and entry, and is a major inducer of neutralizing antibodies. This paper was to find a soluble and functional recombinant receptor-binding domain of severe acute respiratory syndrome-associated coronavirus (SARS-Cov), and to analyze its receptor binding ability. METHODS Three fusion tags (glutathione S-transferase, GST; thioredoxin, Trx; maltose-binding protein, MBP), which preferably contributes to increasing solubility and to facilitating the proper folding of heteroprotein, were used to acquire the soluble and functional expression of RBD protein in Escherichia coli (BL21(DE3) and Rosetta-gamiB(DE3) strains). The receptor binding ability of the purified soluble RBD protein was then detected by ELISA and flow cytometry assay. RESULTS RBD of SARS-Cov spike protein was expressed as inclusion body when fused as TrxA tag form in both BL21 (DE3) and Rosetta-gamiB (DE3) under many different cultures and induction conditions. And there was no visible expression band on SDS-PAGE when RBD was expressed as MBP tagged form. Only GST tagged RBD was soluble expressed in BL21(DE3), and the protein was purified by AKTA Prime Chromatography system. The ELISA data showed that GST/RBD antigen had positive reaction with anti-RBD mouse monoclonal antibody 1A5. Further flow cytometry assay demonstrated the high efficiency of RBD's binding ability to ACE2 (angiotensin-converting enzyme 2) positive Vero E6 cell. And ACE2 was proved as a cellular receptor that meditated an initial-affinity interaction with SARS-Cov spike protein. The geometrical mean of GST and GST/RBD binding to Vero E6 cells were 77.08 and 352.73 respectively. CONCLUSION In this paper, we get sufficient soluble N terminal GST tagged RBD protein expressed in E.coli BL21(DE3); data from ELISA and flow cytometry assay demonstrate that the recombinant protein is functional and binding to ACE2 positive Vero E6 cell efficiently. And the recombinant RBD derived from E.coli can be used to developing subunit vaccine to block S protein binding with receptor and to neutralizing SARS-Cov infection.


title: [Management of corona virus disease-19 (COVID-19): the Zhejiang experience].

text: The current epidemic situation of corona virus disease-19 (COVID-19) still remained severe. As the National Clinical Research Center for Infectious Diseases, the First Affiliated Hospital of Zhejiang University School of Medicine is the primary medical care center for COVID-19 inZhejiang Province. Based on the present expert consensus carried out by National Health Commission and National Administration of Traditional Chinese Medicine, our team summarized and established an effective treatment strategy centered on "Four-Anti and Two-Balance" for clinical practice. The "Four-Anti and Two-Balance"strategy included antivirus, anti-shock, anti-hyoxemia, anti-secondary infection, and maintaining of water, electrolyte and acid base balance and microecological balance. Meanwhile, integrated multidisciplinarypersonalized treatment was recommended to improve therapeutic effect. The importance of early viralogical detection, dynamic monitoring of inflammatory indexes and chest radiograph was emphasized in clinical decision-making. Sputum was observed with the highest positive rate of RT-PCR results. Viral nucleic acids could be detected in10% patients'blood samples at acute periodand 50% of patients had positive RT-PCR results in their feces. We also isolated alive viral strains from feces, indicating potential infectiousness of feces.Dynamic cytokine detection was necessary to timely identifyingcytokine storms and application of artificial liver blood purification system. The "Four-Anti and Two-Balance"strategyeffectively increased cure rate and reduced mortality. Early antiviral treatment could alleviate disease severity and prevent illness progression, and we found lopinavir/ritonavir combined with abidol showed antiviraleffects in COVID-19. Shock and hypoxemia were usually caused by cytokine storms. The artificial liver blood purification system could rapidly remove inflammatory mediators and block cytokine storm.Moreover, it also favoredthe balance of fluid, electrolyte and acid-base and thus improved treatment efficacy in critical illness. For cases of severe illness, early and also short periods of moderate glucocorticoid was supported. Patients with oxygenation index below 200 mmHg should be transferred to intensive medical center. Conservative oxygen therapy was preferred and noninvasive ventilation was not recommended. Patients with mechanical ventilation should be strictly supervised with cluster ventilator-associated pneumonia prevention strategies. Antimicrobial prophylaxis should be prescribed rationally and was not recommended except for patients with long course of disease, repeated fever and elevated procalcitonin (PCT), meanwhile secondary fungal infection should be concerned.Some patients with COVID-19 showed intestinal microbialdysbiosis with decreasedprobiotics such as Lactobacillus and Bifidobacterium. Nutritional and gastrointestinal function should be assessed for all patients.Nutritional support and application of prebiotics or probiotics were suggested to regulate the balance of intestinal microbiota and reduce the risk of secondary infection due to bacterial translocation. Anxiety and fear were common in patients with COVID-19. Therefore, we established dynamic assessment and warning for psychological crisis. We also integrated Chinese medicine in treatment to promote disease rehabilitation through classification methods of traditional Chinese medicine. We optimized nursing process for severe patients to promote their rehabilitation. It remained unclear about viral clearance pattern after the SARS-CoV-2 infection. Therefore, two weeks' quarantine for discharged patients was required and a regular following up was also needed.The Zhejiang experience above and suggestions have been implemented in our center and achieved good results. However, since COVID-19 was a newly emerging disease, more work was warranted to improve strategies of prevention, diagnosis and treatment for COVID-19.


title: Modeling the early events of severe acute respiratory syndrome coronavirus infection in vitro.

text: The clinical picture of severe acute respiratory syndrome (SARS) is characterized by pulmonary inflammation and respiratory failure, resembling that of acute respiratory distress syndrome. However, the events that lead to the recruitment of leukocytes are poorly understood. To study the cellular response in the acute phase of SARS coronavirus (SARS-CoV)-host cell interaction, we investigated the induction of chemokines, adhesion molecules, and DC-SIGN (dendritic cell-specific ICAM-3-grabbing nonintegrin) by SARS-CoV. Immunohistochemistry revealed neutrophil, macrophage, and CD8 T-cell infiltration in the lung autopsy of a SARS patient who died during the acute phase of illness. Additionally, pneumocytes and macrophages in the patient's lung expressed P-selectin and DC-SIGN. In in vitro study, we showed that the A549 and THP-1 cell lines were susceptible to SARS-CoV. A549 cells produced CCL2/monocyte chemoattractant protein 1 (MCP-1) and CXCL8/interleukin-8 (IL-8) after interaction with SARS-CoV and expressed P-selectin and VCAM-1. Moreover, SARS-CoV induced THP-1 cells to express CCL2/MCP-1, CXCL8/IL-8, CCL3/MIP-1alpha, CXCL10/IP-10, CCL4/MIP-1beta, and CCL5/RANTES, which attracted neutrophils, monocytes, and activated T cells in a chemotaxis assay. We also demonstrated that DC-SIGN was inducible in THP-1 as well as A549 cells after SARS-CoV infection. Our in vitro experiments modeling infection in humans together with the study of a lung biopsy of a patient who died during the early phase of infection demonstrated that SARS-CoV, through a dynamic interaction with lung epithelial cells and monocytic cells, creates an environment conducive for immune cell migration and accumulation that eventually leads to lung injury.


title: Identification of coronaviruses by the use of indirect protein A-gold immunoelectron microscopy.

text: Concentration by airfuge and protein A-colloidal gold immunoelectron microscopy (PAG-IEM) offered a rapid and sensitive method for detection and identification of coronaviruses from various species. The method was applied to partially purified tissue culture-adapted or egg-adapted mammalian and avian coronaviruses and to clarified fecal samples from diarrheic calves and turkey poults for detection of enteric coronaviruses. Aggregates of virus coated with specific antibody were seen in virus samples mixed with homologous antiserum but not in control samples containing preexposure serum. At least a 10-50-fold enhancement of the sensitivity of direct EM for virus detection was obtained using protein A-colloidal gold complex as an electron-dense marker. The PAG-IEM method demonstrated low nonspecific background labeling and permitted detection of soluble and particle-associated antigen. Reciprocal cross-reactivity was detected among the subgroup of mammalian hemagglutinating coronaviruses, and antisera to 4 members of other subgroups only recognized their homologous virus.


title: Malaysia collaborates with the new york academy of sciences to develop an innovation-based economy.

text: If Malaysia is to become a high-income country by 2020, it will have to transform into a knowledge-based, innovation economy. This goal will be achieved by developing an atmosphere conducive to experimentation and entrepreneurship at home; while reaching out to partners across the globe. One of Malaysia's newest partnerships is with the New York Academy of Sciences. The Academy has expertise in innovation and higher education and a long history of promoting science, education, and science-based solutions through a global network of scientists, industry-leaders, and policy-makers. Malaysia's Prime Minister, Dato' Sri Mohd Najib Tun Abdul Razak, leveraged the Academy's network to convene a science, technology, and innovation advisory council. This council would provide practical guidance to establish Malaysia as an innovation-based economy. Three initial focus areas, namely palm-oil biomass utilisation, establishment of smart communities, and capacity building in science and engineering, were established to meet short-term and long-term targets.


title: Serological evidence of coronavirus infection in mithuns (Bos frontalis) from India.

text: Studies conducted on mithuns at the National Research Centre on Mithun, and mithuns found in free-ranging conditions in Nagaland, India, revealed that the overall prevalence of antibodies to coronavirus was 80% (95% confidence interval [CI] 77-83) when detected by a commercially available enzyme-linked immunosorbent assay. The highest seroprevalence (92%; 95% CI 89-95) was found in mithuns above three years of age, and the lowest (53%; 95% CI 36-70) in mithuns of 7 to 12 months old (P = 0.00005). The study also revealed a higher degree of positivity (++++) in mithuns above three years old than in those below one year. No statistically significant difference in seroprevalence was observed between male (80%; 95% CI 75-85) and female mithuns (80%; 95% CI 76-84). The seroprevalence was found to be higher (92%; 95% CI 89-95) in mithuns reared in semi-intensive systems compared with those kept under free-ranging conditions (56%; 95% CI 49-63; P = 0.003). This is probably the first serological evidence of coronavirus infection in mithuns. The possible role of adults in the epidemiology of coronavirus infection in mithuns is also discussed.


title: Wellbeing.

text: We all know a well-balanced diet is important for our general health and wellbeing, but how easy is it to maintain when working rotating shifts and dealing with busy workloads?


title: Artificial oxygen carriers as a possible alternative to red cells in clinical practice.

text: Fluid resuscitation is intended to eliminate microcirculatory disorders and restore adequate tissue oxygenation. The safety limits for a restrictive transfusion policy are given by patients' individual tolerance of acute normovolemic anemia. Artificial oxygen carriers based on perfluorocarbon or hemoglobin are attractive alternatives to allogenic red blood cells. There are many risks involved in allogenic blood transfusions and they include transmission of infections, delayed postoperative wound healing, transfusion reactions, immunomodulation and cancer recurrence. Regardless of whether artificial oxygen carriers are available for routine clinical use, further studies are needed in order to show the safety and efficacy of these substances for clinical practice.


title: Solitary Fibrous Tumour A rare cause of acute abdomen.

text: Solitary Fibrous Tumours(SFT), previously known as haemangiopericytoma, are rarely encountered in surgery. They arise from mesenchyme tissue and can occur at several sites in the body - head and neck, extremities, thorax, abdomen and retroperitoneal space. In the thorax, where they arise from the pleura, and abdomen they may attain a large size before giving rise to symptoms. Most SFT behave in a benign manner. However a number of them recur locally or metastasize. Recurrences can occur several years after excision of the primary tumour. Complete surgical excision remains the primary modality of treatment. But, in sites where complete excision is not possible, other modalities have been tried with varying success. Here, we describe a SFT of the mesentery of the small intestine, an uncommon manifestation of the tumour, recurring after a period of 19 years in a 55 year old female, and presenting to the Emergency Department as an acute abdomen caused by acute intestinal obstruction. Surgical excision of the tumour was performed together with primary anastomosis of the small intestine. KEY WORDS Acute abdomen, Late recurrence, Mesentery, Solitary Fibrous tumour.


title: The relationship between the occurrence of undifferentiated bovine respiratory disease and titer changes to bovine coronavirus and bovine viral diarrhea virus in 3 Ontario feedlots.

text: Serological evidence of previous viral exposure (titer at arrival) and current viral exposure (titer increase) during a 28-day study period, was used to determine if bovine coronavirus (BCV) or bovine viral diarrhea virus (BVDV) was associated with the occurrence of undifferentiated bovine respiratory disease (UBRD) in feedlot calves. Neutralizing antibody titers to BCV and BVDV were determined for 852 animals from 3 Ontario feedlots. Calves at 2 of the 3 feedlots (n = 753) received a modified live 4-way viral vaccine containing BVDV. On arrival at the feedlots, 90% of animals were seropositive for BCV, while 39% of animals were seropositive for BVDV. This evidence of previous exposure to both viruses was associated with reduced subsequent UBRD risk. Evidence of exposure to BCV during the study period was common, as 50% of animals showed a 16-fold or greater titer increase; however, treatment for UBRD was not associated with titer change. Although the majority of animals were vaccinated for BVDV at arrival, within a feedlot, animals treated for UBRD had larger titer increases to BVDV than non-treated animals. Based on our findings we infer that BCV was not causally related to UBRD occurrence, however consistent with other literature, BVDV may be causally related to UBRD occurrence.


title: Nanotechnology: a new frontier in virus detection in clinical practice.

text: Researchers are expanding the applications of nanotechnology in the field of medicine since mid-2000. These technologies include nanoarrays, protein arrays, nanopore technology, nanoparticles as a contrivance in immunoassays and nanosensors, among others. Nanobiotechnologies are clinically applicable and possess the potential to be useful in laboratory diagnosis of infections in general and viral infections in particular. Nanotechnology is a significant advance in molecular diagnostics. The technology strengthens and expands the DNA and protein microarray methods. In particular, the waveguide technology is an emergent area with many diagnostic applications. Nanosensors are the new contrivance for detection of bioterrorism agents. All these new technologies would have to be evaluated in clinical settings before their full import is appreciated and accepted.


title: Infectious RNA transcribed in vitro from a cDNA copy of the human coronavirus genome cloned in vaccinia virus.

text: The coronavirus genome is a positive-strand RNA of extraordinary size and complexity. It is composed of approximately 30000 nucleotides and it is the largest known autonomously replicating RNA. It is also remarkable in that more than two-thirds of the genome is devoted to encoding proteins involved in the replication and transcription of viral RNA. Here, a reverse-genetic system is described for the generation of recombinant coronaviruses. This system is based upon the in vitro transcription of infectious RNA from a cDNA copy of the human coronavirus 229E genome that has been cloned and propagated in vaccinia virus. This system is expected to provide new insights into the molecular biology and pathogenesis of coronaviruses and to serve as a paradigm for the genetic analysis of large RNA virus genomes. It also provides a starting point for the development of a new class of eukaryotic, multi-gene RNA vectors that are able to express several proteins simultaneously.


Contrasting experiences of state public health law reform pursuant to the Turning Point Model State Public Health Act.


title: Characteristics of patients with COVID-19 pneumonia at Hvidovre Hospital, March-April 2020.

text: INTRODUCTION The first case of coronavirus disease 2019 (COVID-19) disease caused by severe acute respiratory syndrome coronavirus-2 occurred in Denmark on 27 February 2020. On 10 March, the first case of COVID-19 pneumonia was admitted to Hvidovre Hospital. METHODS Retrospective case review of individuals 18 years or older who were admitted consecutively to Hvidovre Hospital from 10 March through 23 April 2020. RESULTS A total of 175 individuals were admitted with COVID-19 pneumonia. The median age was 71 years, 48.6% were male and 71% had at least one co-morbidity. The most commonly presenting symptoms were dyspnoea, dry cough, and fever. The majority of patients had lymphopenia, elevated liver function tests and C-reactive protein. Nearly two in three presented with multilobar infiltration by chest X-ray. Respiratory failure leading to invasive mechanical ventilation developed in 27 patients (15.4%). By 20 April, 23 of 175 (13.1%) patients remained hospitalised, 43 (24.6%) had died and 109 (62.3%) had been discharged. CONCLUSIONS The manifestations of COVID-19 at presentation were similar to those seen in other reports. Our population was older, slightly overrepresented by women and had a high level of co-morbidity. COVID-19 admittance was associated with frequent need of intensive care and mechanical ventilation that was associated with a very high mortality. FUNDING none. TRIAL REGISTRATION not relevant.


title: Construction, characterization, and immunogenicity of an attenuated Salmonella enterica serovar typhimurium pgtE vaccine expressing fimbriae with integrated viral epitopes from the spiC promoter.

text: Transmissible gastroenteritis virus (TGEV) is a porcine coronavirus that causes diarrhea, leading to near 100% mortality in neonatal piglets with corresponding devastating economic consequences. For the protection of neonatal and older animals, oral live vaccines present the attractive property of inducing desired mucosal immune responses, including colostral antibodies in sows--an effective means to passively protect suckling piglets. Newly attenuated Salmonella vaccine constructs expressing TGEV S protein epitopes were studied and evaluated for improved humoral immune response to TGEV. The macrophage-inducible Salmonella ssaH and spiC/ssaB promoters were compared for their ability to express the TGEV C and A epitopes in the context of the heterologous 987P fimbriae on Salmonella vaccines. Compared to the ssaH promoter, the Salmonella cya crp vector elicited significantly higher levels of mucosal and systemic antibodies in orally immunized mice when the chimeric fimbriae were expressed from the spiC promoter. The Salmonella spiC promoter construct induced the highest level of chimeric fimbriae after being taken up by the J774A.1 macrophagelike cells. The Salmonella cya crp vaccine vector was shown to incorporate into 987P partially degraded chimeric subunits lacking the TGEV epitopes. In contrast, its isogenic pgtE mutant produced fimbriae consisting exclusively of intact chimeric subunits. Mice immunized orally with the Salmonella pgtE vaccine expressing chimeric fimbriae from the spiC promoter elicited significantly higher systemic and mucosal antibody titers against the TGEV epitopes compared to the parental vaccine. This study indicates that the Salmonella cya crp pgtE vector and the spiC promoter can be used successfully to improve immune responses toward heterologous antigens.


title: RNA-protein interactions: involvement of NS3, NS5, and 3' noncoding regions of Japanese encephalitis virus genomic RNA.

text: The mechanism of replication of the flavivirus Japanese encephalitis virus (JEV) is not well known. The structures at the 3' end of the viral genome are highly conserved among divergent flaviviruses, suggesting that they may function as cis-acting signals for RNA replication and, as such, might specifically bind to cellular or viral proteins. UV cross-linking experiments were performed to identify the proteins that bind with the JEV plus-strand 3' noncoding region (NCR). Two proteins, p71 and p110, from JEV-infected but not from uninfected cell extracts were shown to bind specifically to the plus-strand 3' NCR. The quantities of these binding proteins increased during the course of JEV infection and correlated with the levels of JEV RNA synthesis in cell extracts. UV cross-linking coupled with Western blot and immunoprecipitation analysis showed that the p110 and p71 proteins were JEV NS5 and NS3, respectively, which are proposed as components of the RNA replicase. The putative stem-loop structure present within the plus-strand 3' NCR was required for the binding of these proteins. Furthermore, both proteins could interact with each other and form a protein-protein complex in vivo. These findings suggest that the 3' NCR of JEV genomic RNA may form a replication complex together with NS3 and NS5; this complex may be involved in JEV minus-strand RNA synthesis.


title: Prevalence and awareness of HIV infection among men who have sex with men --- 21 cities, United States, 2008.

text: Men who have sex with men (MSM) are at increased risk for infection with human immunodeficiency virus (HIV). In 2006, 57% of new HIV infections in the United States occurred among MSM. To estimate and monitor risk behaviors, CDC's National HIV Behavioral Surveillance system (NHBS) collects data from metropolitan statistical areas (MSAs) using an anonymous cross-sectional interview of men at venues where MSM congregate, such as bars, clubs, and social organizations. This report summarizes NHBS data from 2008, which indicated that, of 8,153 MSM interviewed and tested in the 21 MSAs participating in NHBS that year, HIV prevalence was 19%, with non-Hispanic blacks having the highest prevalence (28%), followed by Hispanics (18%), non-Hispanic whites (16%), and persons who were multiracial or of other race (17%). Of those who were infected, 44% were unaware of their infection. Men who know their current HIV infection status can be linked to appropriate medical care and prevention services. Once linked to prevention services, men can learn ways to avoid transmitting the virus to others. Young MSM (aged 18--29 years) (63%) and minority MSM (other than non-Hispanic white) (54%) were more likely to be unaware of their HIV infection. Efforts to ensure at least annual HIV testing for MSM should be strengthened, and HIV testing and prevention programs should increase their efforts to reach young and minority MSM.


Breath of life.


title: Antibodies to coronaviruses OC43 and 229E in multiple sclerosis patients.

text: Multiple sclerosis (MS) and matched control sera had similar antibody titers to coronaviruses OC43 and 229E when tested by a radioimmunoassay method. In contrast, cerebrospinal fluid from MS patients contained coronavirus antibodies more frequently and in higher titers than matched controls. Intrathecal antibody synthesis to OC43 and 229E viruses was detected in 41% (9/22) and 26% (7/27) of MS patients, respectively, but was not found in any of the neurologic control patients. This intrathecal antibody synthesis may mean that coronaviruses play an etiologic or pathogenic role in MS. Alternatively, intrathecal synthesis of coronavirus antibodies may be but part of a generalized and variable intrathecal antibody synthesis that is typical for MS patients.


title: Viral haemagglutination of glutaraldehyde-fixed sheep erythrocytes.

text: Sheep erythrocytes stabilized by treatment with glutaraldehyde were used successfully in haemagglutination tests with Newcastle disease, and equine influenza viruses. The fixation enabled storage of the cells for at least 21 days without altering their agglutinable properties. Stability was best when the cells were stored at -114 degrees C in the vapour phase of liquid nitrogen.


title: Binding of the influenza A virus to cell-surface receptors: structures of five hemagglutinin-sialyloligosaccharide complexes determined by X-ray crystallography.

text: The structures of five complexes of the X-31 influenza A (H3N2) virus hemagglutinin with sialyloligosaccharide receptor analogs have been determined from 2.5 to 2.8 A resolution by X-ray crystallography. There is well-defined electron density for three to five saccharides in all five complexes and a striking conformational difference between two linear pentasaccharides with the same composition but different linkage [alpha(2-->6) or alpha(2-->3)] at the terminal sialic acid. The bound position of the terminal sialic acid (NeuAc) is the same in all five complexes and is identical to that reported previously from the study of mono- and trisaccharides. The two oligosaccharides with NeuAc alpha(2-->6)Gal linkages and GlcNAc at the third position have a folded conformation with the GlcNAc doubled back to contact the sialic acid. The pentasaccharide with a terminal NeuAc alpha(2-->3)Gal linkage and GlcNAc at the third position has an extended (not folded) conformation and exits from the opposite side of the binding site than the alpha(2-->6)-linked molecule of the same composition. The difference between the conformation of the pentasaccharide with a 2,6 linkage and the trisaccharide 2,6-sialyllactose suggests that 2,6-sialyllactose is not, as previously believed, an appropriate analog of natural influenza A virus receptors. The oligosaccharides studied are NeuAc alpha(2-->3)Gal beta(1-->4)Glc, NeuAc alpha(2-->6)Gal beta(1-->4)Glc, NeuAc alpha(2-->3)Gal beta(1-->3)GlcNAc beta(1-->3)Gal beta(1-->4)Glc, NeuAc alpha(2-->6)Gal beta(1-->4)GlcNAc beta(1-->3)Gal beta(1-->4)Glc, and [NeuAc alpha(2-->6)Gal beta(1-->4)GlcNAc]2 beta(1-->3/6)Gal-beta-O-(CH2)5-COOCH3.


title: De novo initiation of viral RNA-dependent RNA synthesis.

text: RNA viruses use several initiation strategies to ensure that their RNAs are synthesized in appropriate amounts, have correct termini, and can be translated efficiently. Many viruses with genomes of single-stranded positive-, negative-, and double-stranded RNA initiate RNA synthesis by a de novo (primer-independent) mechanism. This review summarizes biochemical features and variations of de novo initiation in viral RNA replication.


title: Full robot-assisted gastrectomy: surgical technique and preliminary experience from a single center.

text: The treatment of gastric cancer requires a multidisciplinary approach in which surgery plays the main role. The diffusion of minimally invasive surgery for gastric cancer treatment is limited by the complexity of performing an extended lymphadenectomy. This surgical step can be facilitated through the use of a robot-assisted system. To date, there are few published articles discussing a full robotic approach that precisely show the different surgical steps. The aim of this study is to describe our experience, surgical techniques and the short-term results of a consecutive series of full robotic gastrectomies using the Da Vinci Surgical System. From November 2011 to January 2015, 17 patients with gastric cancer underwent curative resection by robotic approach for locally advanced tumors. In summary, there were 15 total gastrectomies with a Roux-en-Y esophagojejunostomy, one total gastrectomy with transverse colectomy and one sub-total gastrectomy. Resection margins were negative in all cases. Conversions occurred in two patients. Robot-assisted gastrectomy with extended lymphadenectomy is a safe technique and successfully allows an adequate lymph node harvest and optimal R0-resection rates with low postoperative morbidity. The learning curve appears to be shorter than in laparoscopic surgery. Further follow-up and randomized clinical trials are required to confirm the role of a robotic approach in gastric cancer surgery.


title: [SARS-CoV-2 infection with gastrointestinal symptoms as the first manifestation in a neonate].

text: Since December 2019, the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has occurred in Wuhan, Hubei Province, China. The infected cases were noted mostly in adults, but rarely reported in children, especially neonates. Most children with SARS-CoV-2 infection present mainly with respiratory symptoms, but less commonly with gastrointestinal symptoms, and tend to have mild clinical symptoms. A neonate with SARS-CoV-2 infection, who had vomiting and milk refusal as the first symptom, was recently admitted to Wuhan Children's Hospital. After two weeks of treatment, the patient recovered gradually and was discharged. Here, this case is reported to improve the understanding of SARS-CoV-2 infection in neonates.