title: Nucleotide sequence and expression of the spike (S) gene of canine coronavirus and comparison with the S proteins of feline and porcine coronaviruses.

text: We have cloned, sequenced and expressed the spike (S) gene of canine coronavirus (CCV; strain K378). Its deduced amino acid sequence has revealed features in common with other coronavirus S proteins: a stretch of hydrophobic amino acids at the amino terminus (the putative signal sequence), another hydrophobic region at the carboxy terminus (the membrane anchor), heptad repeats preceding the anchor, and a cysteine-rich region located just downstream from it. Like other representatives of the same antigenic cluster (CCV-Insavc-1 strain, feline infectious peritonitis and enteric coronaviruses, porcine transmissible gastroenteritis and respiratory coronaviruses, and the human coronavirus HCV 229E), the CCV S polypeptide lacks a proteolytic cleavage site present in many other coronavirus S proteins. Pairwise comparisons of the S amino acid sequences within the antigenic cluster demonstrated that the two CCV strains (K378 and Insavc-1) are 93.3% identical, about as similar to each other as they are to the two feline coronaviruses. The porcine sequences are clearly more divergent mainly due to the large differences in the amino-terminal (residues 1 to 300) domains of the proteins; when only the carboxy-terminal parts (residues 301 and on) are considered the homologies between the canine, feline and porcine S polypeptides are generally quite high, with identities ranging from 90.8% to 96.8% . The human coronavirus is less related to the other members of the antigenic group. A phylogenetic tree constructed on the basis of the S sequences showed that the two CCVs are evolutionarily more related to the feline than to the porcine viruses. Expression of the CCV S gene using the vaccinia virus T7 RNA polymerase system yielded a protein of the expected M(r) (approximately 200K) which could be immunoprecipitated with an anti-feline infectious peritonitis virus polyclonal serum and which was indistinguishable from the S protein synthesized in CCV-infected cells.


title: SARS-CoV nucleocapsid protein induced apoptosis of COS-1 mediated by the mitochondrial pathway.

text: To investigate the apoptosis effect of SARS coronavirus nucleocapsid protein on cultured cell lines and to explore the possible pathway of apoptosis. pCDNA3.1(-)/his-myc vector containing the SARS coronavirus nucleocapsid gene (N), matric gene (M), spike gene (S) were transfected into COS-1, Huh-7 and HepG2 cells. Apoptosis induced by SARS coronavirus N protein under starvation of serum of COS-1 cells was monitored by Annexin V and electron microscopy assays. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (DeltaPsim) were determined by flow cytometric assay. Cytochrome C, cleaved caspase (cysteine aspartic acid protease)-3, 9, and poly (ADP-ribose) polymerase (PARP) were detected by Western blot. After removal of serum in COS-1 cells, we observed the loss of DeltaPsim, the increase of ROS and cytochrome C release into cytosol and subsequent activation of caspase-3 and PARP cleavage. The pan-caspase inhibitor z-VAD-fmk can block the activation of caspase 3, 9 and PARP cleavage. In conclusion, SARS coronavirus N protein can induce apoptosis of COS-1 cells by activating mitochondrial pathway. SARS coronavirus M, S protein can not induce apoptosis in COS-1, HepG2 and Huh-7 and SARS coronavirus N protein can not induce apoptosis in HepG2 and Huh-7 by methods used in this study.


title: [Protective effect of specific antibody in serum of convalescent patient with SARS].

text: OBJECTIVE To investigate inhibitory effect of serum severe acute respiratory syndrome (SARS) -specific antibodies from convalescent patients after half an year of onset on SARS-CoV-mediated cytopathic response. METHODS SARS-CoV immunoglobulin G (IgG) antibody was determined by enzyme linked immunoadsorbent assay (ELISA). Twelve serum samples from convalescent patients, diluted by 1:8 with maintenance medium, were mixed with the three dilution supernatants of SARS-CoV. SARS-CoV were isolated, cultured and identified by the Guangzhou Institute of Respiratory Disease, and cultured with Vero E6 cell suspension. The extent of cytopathic response was observed. RESULTS The absorbance (A) value of SARS-CoV IgG antibody ranged from 0.81 to 2.06 in patients after half an year of SARS onset, and form 0.79 to 2.01 in patients before half an year of SARS onset. The extent of cytopathic response was decreased by more than 25% in all 12 convalescent patients, as compared with control serum. CONCLUSION The A value of SARS-CoV IgG antibody in serum of convalescent patients tended to elevate in half an year after SARS onset. SARS-CoV IgG antibody could inhibit SARS-CoV-mediated cytopathic response, indicating it might be one of protective antibodies.


title: The development, optimization and validation of an assay for high throughput antiviral drug screening against Dengue virus.

text: Dengue virus (DENV) is listed as one of the NIAID Category A priority pathogens. Dengue disease is endemic in most tropical countries, with an estimated 2.5 billion people living in areas at risk of DENV infection. Due to the lack of vaccines and antiviral drugs, it is now a huge public health burden around the world. In order to screen large compound libraries for the identification of novel antivirals targeting DENV, it is essential to develop a high throughput screening (HTS) amenable assay. Here, we present the development, optimization and validation of a cytopathic effect-based assay against Dengue virus serotype-2 (DENV-2). The assay conditions, including cell culturing conditions, DMSO tolerance and the multiplicity of infection, were optimized in both 96- and 384-well plates. Assay robustness and reproducibility were determined under the optimized conditions in 96-well plate, including Z'-value of 0.71, signal-to-background ratio of 6.88, coefficient of variation of 6.3% in mock-infected cells and 12.3% in DENV-2 infected cells. This assay was further miniaturized into a 384-well plate format with similar assay robustness and reproducibility comparing with these in the 96-well plate format. This assay was then validated using the LOPAC(1280) compound library, demonstrating its repeatability with comparable assay robustness and reproducibility. This fully developed and validated HTS amenable assay could be used in future studies to screen large compound libraries for the identification of novel antivirals against dengue disease.


title: Use of a reverse-transcriptase polymerase chain reaction for monitoring the shedding of feline coronavirus by healthy cats.

text: The pattern of shedding of feline coronavirus (FCoV) was established in 155 naturally infected pet cats from 29 households over periods of up to five years. Viral RNA was detected in faeces by reverse-transcriptase PCR (RT-PCR), and plasma antiviral antibodies by immunofluorescence. The cats rarely shed FCoV in their saliva. Three patterns of FCoV shedding were observed. Eighteen of the cats shed virus continuously, so were persistent, and possibly lifelong, carriers; none of them developed feline infectious peritonitis. Fifty-six cats ceased shedding virus, although they were susceptible to reinfection, and 44 shed intermittently or were being continuously reinfected. Four of the cats were resistant to infection. Seventy-three per cent of the virus shedding episodes lasted up to three months and 95 per cent up to nine months. There was a correlation between shedding and antibody titre but the cats could remain seropositive for some time after they had ceased shedding virus. One-off testing for FCoV by RT-PCR is inappropriate. Identification of longterm carriers requires that a positive result be obtained by RT-PCR on faecal samples for at least eight consecutive months. A cat should be shown to be negative over five months, or to have become seronegative, to ensure that it has ceased shedding virus.


title: Neurological manifestations in severe acute respiratory syndrome.

text: During the worldwide outbreak of severe acute respiratory syndrome (SARS) in 2002-2003, there were 664 probable SARS patients reported in Taiwan. SARS patients usually present with symptoms related to the respiratory system while neurological manifestations have rarely been described. There were three patients who developed axonopathic polyneuropathy 3-4 weeks after onset of SARS; their clinical condition and electrophysiological studies revealed obvious improvement at follow-up. Two SARS patients have experienced myopathy and three other patients developed rhabdomyolysis. These neuromuscular disorders in SARS patients were considered as critical illness neuropathy and myopathy, but the possibility of direct attack by SARS coronavirus on the nerve and muscle could not be excluded. Large artery ischemic stroke were described in five SARS patients with poor prognosis. Multiple factors contributed to this vascular insult included hypercoagulabe status related to both SARS coronavirous and the usage of intravenous immunoglobulin, septic and cardiogenic shock, and possible vasculitis. The relationship between SARS and above neurological problems still needs further clarification. Pathological and microbiological studies are mandatory to delineate this issue.


title: Palpable Fear: COVID-19's Toll on Physician Morale and Mental Health.

text: As COVID-19 ravaged human lives, it also took a toll on physician morale and mental health.


title: Protection studies on winter dysentery caused by bovine coronavirus in cattle using antigens prepared from infected cell lysates.

text: Cells infected with bovine coronavirus (BCV) were solubilized with Triton X-100 to yield a cell lysate (CL) antigen having high hemagglutinating (HA) titers. The antigen gave high HA titers using rat erythrocytes, suggesting that it contained large amounts of hemagglutinin esterase (HE) antigen. The CL antigen, combined with an oil adjuvant, was tested for protective and antibody-inducing activities in cattle. Four groups (2 cattle/group) of cattle were inoculated with CL antigen having HA titers of 16 000, 4000, 1000, and 250. Another group served as untreated controls. Two intramuscular inoculations were given at an interval of 3 wk. The animals were challenged with virus 1 wk after the second inoculation. The groups immunized with the CL antigen having an HA titer of 4000 or 16 000 produced hemagglutination inhibition (HI) antibody titers of > 320 and serum neutralizing (SN) antibody titers of > 1280. These groups of animals showed no clinical abnormalities after challenge. In the groups immunized with CL antigen at an HA titer of 1000 or 250, HI antibody titers were 40 to 160 and SN titers were 80 to 640. The cattle with HI antibody titers of > or = 160 and the SN titers of > or = 640 showed no clinical signs, but the cattle with the HI antibody titer < 80 and the SN antibody titer < 160 developed watery diarrhea and fever after challenge. These results indicate that CL antigen with high HA titer induces antibody production in cattle that provides effective protection against winter dysentery.


title: Retrospective serological investigation of severe acute respiratory syndrome coronavirus antibodies in recruits from mainland China.

text: Different assays were used to analyze 1,621 serum specimens collected from military recruits from the People's Republic of China in 2002 for severe acute respiratory syndrome (SARS) coronavirus antibodies. The results demonstrated that the subjects either had rarely been exposed to the virus before the 2003 SARS outbreak or had not been exposed but the nucleocapsid protein cross-reacted with other antibodies in humans.


title: Protein nanopatterns and biosensors using gold binding polypeptide as a fusion partner.

text: An efficient strategy for immobilizing proteins on a gold surface was developed by employing the gold binding polypeptide (GBP) as a fusion partner. Using the enhanced green fluorescent protein (EGFP), severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVme), and core streptavidin (cSA) of Streptomyces avidinii as model proteins, specific immobilization of the GBP-fusion proteins onto the gold nanoparticles and generation of protein nanopatterns on the bare gold surface were demonstrated. The GBP-fused SCVme bound to gold nanoparticles successfully interacted with its antibody and showed changes in absorbance and color, allowing efficient diagnosis of SARS-CoV. The fusion proteins could be successfully immobilized on the gold surface by nanopatterning and microcontact printing as examined by atomic force microscopy and surface plasmon resonance analysis. The poly(dimethylsiloxane) microfluidic channels were created on the gold surface and were used for antigen-antibody and DNA-DNA interaction studies. Specific immobilization of GBP-EGFP fusion protein and its interaction with the antibody in the microchannels could be demonstrated. By immobilizing the DNA probe through the use of GBP-fused cSA, specific hybridization of the target DNA prepared from Salmonella could also be achieved. The GBP-fusion method allows immobilization of proteins onto the gold surface without surface modification and in bioactive forms suitable for studying protein-protein, DNA-DNA, and other biomolecular interaction studies. Furthermore, these studies can be carried out in a microfluidic system, which allows high-throughput analysis of biomolecular interactions.


title: Life-threatening human metapneumovirus pneumonia requiring extracorporeal membrane oxygenation in a preterm infant.

text: We present the first report in the literature of a child with human metapneumovirus pneumonia who required extracorporeal membrane oxygenation for survival. This was a 3-month-old premature boy from British Columbia, Canada, who developed severe respiratory failure, experienced failure of high-frequency oscillatory mechanical ventilation, and required extracorporeal membrane oxygenation support for 10 days. This case illustrates the importance of including this newly discovered pathogen among the causes of childhood pneumonia.


title: [SARS-CoV-2 infection and solid organ transplantation].

text: Transplantation has become a valid therapeutic option for an increasing number of patients with end-stage organ disease. The emergence of SARS-CoV-2 coronavirus infection and associated disease (COVID-19) has alarmed the transplant community, since recommendations for adequate follow-up of organ transplant recipients during the acute phase of a pandemic are limited. Furthermore, treatment options against COVID-19 disease and adequate adjustment of immunosuppression in at risk patients remain a concern. This review summarizes current knowledge on the incidence and clinical course of SARS-CoV-2 infection in patients with solid organ transplantation. It also discusses therapeutic strategies and provides general recommendations on how to proceed with transplantation programs in a time when health care resources may become scarce.


title: Incidence and surgical treatment of synthetic mesh-related infectious complications after laparoscopic ventral rectopexy.

text: BACKGROUND Prosthetic-related infection and erosion occurring after a laparoscopic ventral rectopexy (LVR) are rare complications, and their importance is often underestimated. The aim of this study was to compare the incidence rate and surgical management of these complications in LVR patients with polyester (PE) or polypropylene (PP) prostheses. METHODS From January 2004 to June 2012, 149 patients underwent LVR with PE and 176 underwent LVR with PP. Surgical management and rate of infectious and erosive prosthesis-related complications, depending on the type of prosthesis, were described and compared. Functional results after complications were assessed. RESULTS Five patients from the PE prosthesis group (3.3 %), compared with two patients from the PP prosthesis group (1.1 %), experienced prosthesis-related infection or erosion (p = 0.16). The rate of erosion alone was 3.3 % in patients with a PE prosthesis, and 0.55 % in patients with a PP prosthesis (p = 0.06). The average time until clinical diagnosis of a prosthesis-related complication was identical for both groups: 31 months (range 3-62 months). All patients underwent surgical removal of the prosthesis: For the five patients from the PE group, complete removal was performed by laparoscopy associated with a transanal procedure. For the two patients in the PP mesh group, laparoscopy was ineffective in removing the mesh which was partially removed through a subsequent transanal procedure. None of the patients had a protective stoma, and in all patients the complication had resolved 12 months after removal. Only one patient had worsening functional symptoms (fecal incontinence) after prosthesis removal. CONCLUSIONS When a prosthesis-related infection or erosion occurs, treatment consists in the surgical removal of the prosthesis by laparoscopy/and/or a transanal procedure. Functional symptoms do not routinely recur after prosthesis removal.


title: Identification of the coronavirus MHV-JHM mRNA 4 product.

text: A bacterial expression vector was constructed to encode a fusion protein which had, at its carboxy terminus, a polypeptide encoded within the 5' proximal open reading frame of the coronavirus MHV-JHM mRNA 4. This polypeptide was isolated and used to produce an antiserum. The antiserum reacted specifically with a 15,000 Mr polypeptide synthesized in MHV-JHM-infected cells, or in vitro translations of infected cell poly(A) RNA enriched for mRNA 4. These results demonstrate the translational activity of mRNA 4 during infection, identify conclusively the translation product and provide a means to investigate the synthesis and function of this protein.


Global impact of severe acute respiratory syndrome: measures to prevent importation into Saudi Arabia.


Protein microarrays.


The Hong Kong SARS report: an Australian perspective.


title: Transoral stapling for Zenker diverticulum: effect of the traction suture-assisted technique on long-term outcomes.

text: BACKGROUND Long-term outcomes and predictors of success after transoral stapling for Zenker diverticulum are still unclear. METHODS Between 2001 and 2010, 91 patients with Zenker diverticulum underwent transoral stapling under general anesthesia. Since 2008, the technique was modified by applying traction sutures to ease engagement of the common septum inside the stapler jaws. Perioperative variables, distribution of symptoms, and outcome of surgery were analyzed. Long-term results were compared between patients undergoing standard versus modified technique of transoral stapling. RESULTS The transoral approach was successfully completed in 79 (86.8 %) patients with a median age of 74 years. Overall morbidity was 5 %, and there was no mortality. The median length of hospital stay was 2 days. Six patients were lost to follow-up. After a median follow-up of 53 (range, 12-114) months, an improvement of dysphagia and regurgitation scores (p < 0.001) and a reduction in the number of pneumonia episodes per year (p < 0.001) was recorded. The long-term success rate of the procedure was 80.1 %. At a median time of 12 months, 14 patients complained of recurrent symptoms, 7 of whom needed an open (n = 4) or transoral (n = 3) reoperation. Use of traction sutures resulted in a greater long-term success compared with the standard procedure (p = 0.04). CONCLUSIONS Transoral stapling is a safe and effective technique. A repeat procedure is feasible in case of recurrent diverticulum. The use of traction sutures applied at the apex of the common septum before stapling might increase the long-term success of the technique.


title: Artificial Intelligence-Powered Search Tools and Resources in the Fight Against COVID-19.

text: Emerging technologies are set to play an important role in our response to the COVID-19 pandemic. This paper explores three prominent initiatives: COVID-19 focused datasets (e.g., CORD-19); Artificial intelligence-powered search tools (e.g., WellAI, SciSight); and contact tracing based on mobile communication technology. We believe that increasing awareness of these tools will be important in future research into the disease, COVID-19, and the virus, SARS-CoV-2.


title: [Characterization of human coronavirus 229E infection among patients with respiratory symptom in Beijing, Oct-Dec, 2007].

text: OBJECTIVE To know the etiology, prevalence, clinical symptoms associated with the infection of the HCoV-229E in the respiratory specimens sampled from adult patients in Beijing. METHODS 158 nasopharyngeal swab specimens were collected from adult patients with fever in Beijing between October and December, 2007. We performed the screening of HCoV-229E by real-time RT-PCR and sequencing of HCoV-229E gene fragments derived from conventional PCR. At meantime, we also screened the HCoV-229E positive samples for the co-infection with HCoV-NL63, HCoV-HKU1 and HMPV by real-time RT-PCR. Finally, demographic and clinical data associated with HCoV-229E infection were examined retrospectively. RESULTS We detected 103 (62.5%) of 158 specimens were positive for HCoV-229E by real-time RT-PCR. When tested for other respiratory viruses, 26 HCoV-229E positive patients were found to be co-infected with other viruses. Of which HCoV-NL63 was observed in 3 specimens (11.5%), HCoV-HKU1 in 3 (11.5%) and HMPV in 20 (76.9%). The main clinical manifestations were noted as: headache (in 70.9%), sore throat (69%), chills (68%), cough (33%), sputum (21.3%), rhinorrhea (21.4%), nasal obstruction (16.5%), and a few of patients were visible as vomiting (6.8%), dyspnea (3.9%), diarrhea (in 1.9%). The rate of HCoV-229E infection in adult patients was found no relative with age and gender. CONCLUSION Our data showed that HCoV-229E is a common and important pathogen in adult patients with acute respiratory symptoms but usually resulted in milder influenza-like illnesses. There might have a local outbreak of HCoV-229E infection in Beijing, Oct-Dec, 2007.


Emerging diseases fuel health screening.


title: Severe acute respiratory syndrome: radiographic appearances and pattern of progression in 138 patients.

text: PURPOSE To retrospectively evaluate the radiographic appearances and pattern of progression of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS Chest radiographs obtained at clinical presentation and during treatment in 138 patients with confirmed SARS (66 men, 72 women; mean age, 39 years; age range, 20-83 years) were assessed. Radiographic appearances of pulmonary parenchymal abnormality, distribution, and extent of involvement on initial chest radiographs were documented. Recognizable patterns of radiographic progression were determined by comparing the overall mean percentage of lung involvement for each patient on serial radiographs. RESULTS Initial chest radiographs were abnormal in 108 of 138 (78.3%) patients and showed air-space opacity. Lower lung zone (70 of 108, 64.8%) and right lung (82 of 108, 75.9%) were more commonly involved. In most patients, peripheral lung involvement was more common (81 of 108, 75.0%). Unifocal involvement (59 of 108, 54.6%) was more common than multifocal or bilateral involvement. No cavitation, lymphadenopathy, or pleural effusion was demonstrated. Four patterns of radiographic progression were recognized: type 1 (initial radiographic deterioration to peak level followed by radiographic improvement) in 97 of 138 patients (70.3%), type 2 (fluctuating radiographic changes) in 24 patients (17.4%), type 3 (static radiographic appearance) in 10 patients (7.3%), and type 4 (progressive radiographic deterioration) in seven patients (5.1%). Initial focal air-space opacity in 44 of 59 patients (74.6%) progressed to unilateral multifocal or bilateral involvement during treatment. CONCLUSION Predominant peripheral location; common progression pattern from unilateral focal air-space opacity to unilateral multifocal or bilateral involvement during treatment; and lack of cavitation, lymphadenopathy, and pleural effusion are the more distinctive radiographic findings of SARS.


title: [Two genotypes of Canine coronavirus simultaneously detected in the fecal samples of healthy foxes and raccoon dogs].

text: 61 fecal samples from healthy foxes and 24 from healthy raccoon dogs were examined by RT-nested PCR assays for the presence and genotypic identification of Canine coronaviruses (CCVs). 77.0% fox samples were recognized as CCV positive, 43 of which belonged to type II and 29 to type I, as well as both genotypes were simultaneously detected in 25 samples. Out of the total 24 fecal samples from raccoon dogs, 22 were CCV positive for type II and 16 for type I. M gene fragments of 8 samples were sequenced, 4 of which were confirmed as CCV type I and the other 4 as CCV type II. Sequence analysis showed that the M gene of CCV type I had a high similarity of 96.7% - 98.1% between the fox-and raccoon dog strains and the reported Italian strain from diarrhea dogs. The two genotypes, with an identity of 88.3% - 89.7%, formed two separate branches in phylogenetic tree. Interestingly, the sequence at several nucleic acid sites of CCV type II differed between foxes and raccoon dogs. The co-existence and popularity of the two CCV genotypes in healthy foxes and raccoon dogs were first confirmed in this article.


title: A note on epidemic models with infective immigrants and vaccination.

text: The roles of immigration and vaccination on disease dynamics are explored in a simple setting that considers the possibility of conferred immunity. We focus on SIR and SIS models with a vaccinated class. Conditions for the existence of multiple endemic steady states and a fold bifurcation are discussed.


title: Identification and phylogenetic diversity of parvovirus circulating in commercial chicken and turkey flocks in Croatia.

text: Phylogenetic diversity of parvovirus detected in commercial chicken and turkey flocks is described. Nine chicken and six turkey flocks from Croatian farms were tested for parvovirus presence. Intestinal samples from one turkey and seven chicken flocks were found positive, and were sequenced. Natural parvovirus infection was more frequently detected in chickens than in turkeys examined in this study. Sequence analysis of 400 nucleotide fragments of the nonstructural gene (NS) showed that our sequences had more similarity with chicken parvovirus (ChPV) (92.3%-99.7%) than turkey parvovirus (TuPV) (89.5%-98.9%) strains. Phylogenetic analysis grouped our sequences in two clades. Also, the higher prevalence of ChPV than TuPV in tested flocks was defined. The necropsy findings suggested a malabsorption syndrome followed by a preascitic condition. Further research of parvovirus infection, pathogenesis, and the possibility of its association with poult enteritis and mortality syndrome (PEMS) and runting and stunting syndrome (RSS) is needed to clarify its significance as an agent of enteric disease.


title: Cytokine responses in severe acute respiratory syndrome coronavirus-infected macrophages in vitro: possible relevance to pathogenesis.

text: The pathogenesis of severe acute respiratory syndrome (SARS) remains unclear. Macrophages are key sentinel cells in the respiratory system, and it is therefore relevant to compare the responses of human macrophages to infections with the SARS coronavirus (SARS-CoV) and other respiratory viruses. Primary human monocyte-derived macrophages were infected with SARS-CoV in vitro. Virus replication was monitored by measuring the levels of positive- and negative-strand RNA, by immunofluorescence detection of the SARS-CoV nucleoprotein, and by titration of the infectious virus. The gene expression profiles of macrophages infected with SARS-CoV, human coronavirus 229E, and influenza A (H1N1) virus were compared by using microarrays and real-time quantitative reverse transcriptase PCR. Secreted cytokines were measured with an enzyme-linked immunosorbent assay. SARS-CoV initiated viral gene transcription and protein synthesis in macrophages, but replication was abortive and no infectious virus was produced. In contrast to the case with human coronavirus 229E and influenza A virus, there was little or no induction of beta interferon (IFN-beta) in SARS-CoV-infected macrophages. Furthermore, SARS-CoV induced the expression of chemokines such as CXCL10/IFN-gamma-inducible protein 10 and CCL2/monocyte chemotactic protein 1. The poor induction of IFN-beta, a key component of innate immunity, and the ability of the virus to induce chemokines could explain aspects of the pathogenesis of SARS.


title: T-lymphocyte profiles in FIV-infected wild lions and pumas reveal CD4 depletion.

text: Feline immunodeficiency virus (FIV) is a lentivirus related to human immunodeficiency virus (HIV) that causes feline AIDS in the domestic cat (Felis catus). Serological surveys indicate that at least 25 other species of cat possess antibodies that cross-react with domestic cat FIV. Most infected nondomestic cat species are without major symptoms of disease. Long-term studies of FIV genome variation and pathogenesis reveal patterns consistent with coadaptation of virus and host in free-ranging FIV-Ple-infected African lions (Panthera leo) and FIV-Pco-infected pumas (Puma concolor) populations. This report examined correlates of immunodeficiency in wild and captive lions and pumas by quantifying CD5(+), CD4(+), and CD8(+) T-cell subsets. Free-ranging FIV-Ple-infected lions had immunofluorescence flow cytometry (IFC) profiles marked by a dramatic decline in CD4(+) subsets, a reduction of the CD4(+)/CD8(+) ratio, reduction of CD8(+)beta(high) cells, and expansion of the CD8(+)beta(low) subset relative to uninfected lions. An overall significant depletion in CD5(+) T-cells in seropositive lions was linked with a compensatory increase in total CD5(-) lymphocytes. The IFC profiles were altered significantly in 50% of the seropositive individuals examined. The FIV-Pco-infected pumas had a more generalized response of lymphopenia expressed as a significant decline in total lymphocytes, CD5(+) T-cells, and CD5(-) lymphocytes as well as a significant reduction in CD4(+) T-cells. Like lions, seropositive pumas had a significant decline in CD8(+)beta(high) cells but differed by not having compensatory expansion of CD8(+)beta(low) cells relative to controls. Results from FIV-infected lions and pumas parallel human and Asian monkey CD4(+) diminution in HIV and SIV infection, respectively, and suggest there may be unrecognized immunological consequences of FIV infection in these two species of large cats.


title: Coronavirus replication in the intestinal and respiratory tracts during infection of calves.

text: Coronaviruses have been isolated from cases of neonatal calf diarrhoea in many parts of the world and were revealed in the respiratory tract recently, during a study of calf pneumonia. In experiments on gnotobiotic calves an enteric coronavirus replicated in the nasal epithelium and a respiratory isolate was shed in faeces. The enteric coronavirus prevented subsequent infection with either homologous enteric or the respiratory isolate. A dual tropism for this agent was emphasized by the detection of coronavirus antigen in intestinal and respiratory epithelium of naturally-infected calves.


docid: tnrfquse

title: Environmental factors and seasonal influenza onset in Okayama city, Japan: case-crossover study.

text: Seasonal influenza infection is a major challenge in public health. The term "seasonal influenza" refers to the typical increase in the number of influenza patients in the winter season in temperature zones. However, it is not clear how environmental factors within a single flu season affect influenza infection in a human population. Therefore, we evaluated the effects of temperature and humidity in the 2006-7 flu season on the onset of seasonal influenza using a case-crossover study. We targeted patients who attended one pediatric clinic in Okayama city, Japan and who were diagnosed as being infected with the seasonal influenza virus. Using 2 references (time-stratified and symmetric bidirectional design), we estimated the effects of average temperature and relative humidity from the onset day (lag0) to 10 days before (lag10). The total number of subjects was 419, and their onset days ranged from 26 December 2006 to 30 April 2007. While the onset was significantly associated with lower temperature, relative humidity was not related. In particular, temperatures before the 3-day incubation period had higher-magnitude odds ratios. For example, the odds ratio and 95% confidence interval for average temperature at time lag 8 was 1.12 (1.08-1.17) per 1.0℃ decrease. Low environmental temperature significantly increased the risk of seasonal influenza onset within the 2006-7 winter season.


title: Update: severe acute respiratory syndrome--United States, 2003.

text: CDC continues to work with state and local health departments, the World Health Organization (WHO), and other partners to investigate cases of severe acute respiratory syndrome (SARS). This report updates reported SARS cases worldwide and in the United States, and summarizes changes in travel recommendations for Beijing and Taiwan, where travel advisories have been downgraded to travel alerts.


title: Hepatitis C virus RNA replication occurs on a detergent-resistant membrane that cofractionates with caveolin-2.

text: The mechanism and machinery of hepatitis C virus (HCV) RNA replication are still poorly understood. In this study, we labeled de novo-synthesized viral RNA in situ with bromouridine triphosphate (BrUTP) in Huh7 cells expressing an HCV subgenomic replicon. By immunofluorescence staining using an anti-BrUTP antibody and confocal microscopy, we showed that the newly synthesized HCV RNA was localized to distinct speckle-like structures, which also contain all of the HCV nonstructural (NS) proteins. These speckles are distinct from lipid droplets and are separated from the endoplasmic reticulum (ER), where some HCV NS proteins also reside. Membrane flotation analysis demonstrated that almost all of the NS5A and part of the NS5B proteins and all of the viral RNA were present in membrane fractions which are resistant to treatment with 1% NP-40 at 4 degrees C. They were cofractionated with caveolin-2, a lipid-raft-associated intracellular membrane protein, in the presence or absence of the detergent. In contrast, the ER-resident proteins were detergent soluble. These properties suggest that the membranes on which HCV RNA replication occurs are lipid rafts recruited from the intracellular membranes. The protein synthesis inhibitors cycloheximide and puromycin did not inhibit viral RNA synthesis, indicating that HCV RNA replication does not require continuous protein synthesis. We suggest that HCV RNA synthesis occurs on a lipid raft membrane structure.


docid: nuqjzgfx

title: [Possible unregistered SARS-CoV-2 infection in a young man with anosmia and ageusia].

text: During the SARS-CoV-2 pandemic, testing of suspected cases in Denmark has recently been limited to those who were critically ill. This has left an increasing number of unregistered infections in the society, obscured quantification and impacted lived lives. This case story reports a possible mild SARS-CoV-2 infection in a healthy young man in his twenties. Due to strategic guidelines, he was never tested for SARS-CoV-2, but the medical record was highly suspicious for infection. Besides the well-known symptoms of SARS-CoV-2, this case also reports total anosmia and ageusia.


title: [Prospective clinical study on serum immunity in IgG-antibodies positive patients convalescent from severe acute respiratory syndrome].

text: OBJECTIVE To investigate the serum immunological characteristics in patients convalescent from severe acute respiratory syndrome (SARS). METHODS In the 1 st, 3 rd, 6 th month after their discharge, eg. SARS-IgG, T cell subsets, blood routine, and the blood biochemistry were systemically determined in SARS convalescent patients. RESULTS The SARS-antibodies could be used as the diagnostic evidence. During the 6 months after discharge, the titers of SARS-antibodies were high, but they lowered along with passage of time. At the first recheck, the CD4(+) lymphocyte count was lower than normal level in 55.9% of patients, the CD3(+) lymphocyte count was lower than normal level in 31.2% of patients, and the CD8(+) lymphocyte count was lower than normal level in 14.0% of patients. At the second recheck, the levels of T cell subsets recovered to normal level in the most patients. CONCLUSION T cell subsets, and the number of leukocyte are abnormal in some patients convalescent from SARS. All the indexes examined recover to normal levels half year after discharge. Therefore, it is necessary to follow up the changes in the levels of SARS-antibodies.


title: Pathogenesis of feline infetious peritonitis: pathologic changes and immunofluorescence.

text: Feline infectious peritonitis (FIP) was experimentally induced in FIP virus (FIPV) antibody-positive and antibody-negative kittens after challenge exposure to live-virus aerosol. Seropositive kittens developed antiviral immunofluorescence and lesions more rapidly after challenge exposure than did seronegative kittens. In seropositive kittens, FIPV antigen was present in macrophages and large mononuclear cells in tracheobronchial lymph nodes, lungs, and trachea on postchallenge-exposure day (PCD) 2; in liver and spleen on PCD 3; in kidneys and omentum on PCD 4; and subsequently in nasal turbinates, thoracic and abdominal lymph nodes, thymus, bone marrow, parotid salivary gland, eyes, and brain. Initial antiviral immunofluorescence on PCD 2 coincided with the onset of viremia and vascular lesions. Systemic lesions characterized by perivascular necrotizing pyogranulomatous inflammation, phlebitis and thrombosis, fibrinous serositis, and generalized lymphoid necrosis developed on PCD 3 and 4. Coronavirus-like particles were observed by electron microscopy in cytoplasmic vacuoles or smooth endoplasmic reticulum of degenerating macrophages in inflammatory lesions. In seronegative kittens, antiviral immunofluorescence in tracheobronchial lymph nodes was first detected on PCD 5, and viremia occurred on PCD 6. Systemic necrotizing lesions, comparable with those observed in seropositive kittens on PCD 3 or 4, did not occur in seronegative kittens until PCD 13 or 16. In both groups of kittens, initial viral infection in regional lymphoreticular tissue was followed by viremia and infection of macrophages in reticuloendothelial organs (liver, spleen, lymph nodes) and perivascular locations. The accelerated onset of infection and lesions indicative of an Arthus-type reaction in challenge-exposed seropositive vs seronegative kittens further supports the immune-mediated pathogenesis of FIP.


Update on emerging infections: news from the Centers for Disease Control and Prevention. Emergence of Cryptococcus gattii--Pacific Northwest, 2004-2010.


title: Seroprevalence of avian influenza virus, infectious bronchitis virus, reovirus, avian pneumovirus, infectious laryngotracheitis virus, and avian leukosis virus in Nigerian poultry.

text: Eight poultry farms in Nigeria, including chickens from nine breeder, 14 broiler, 28 pullet, 11 layer, and three cockerel flocks, were tested for antibody seroprevalence to the following poultry viruses of potential economic importance: infectious bronchitis virus (IBV), avian reovirus, avian pneumovirus (APV), infectious laryngotracheitis virus (ILTV), avian influenza virus (AIV), and avian leukosis virus (ALV). Serum samples were collected between 1999 and 2004 and were tested for antibodies using commercial enzyme-linked immunosorbent assay (ELISA) kits. Seroprevalence was very high for IBV (84%); intermediate for reovirus (41%), APV (40%), and ILTV (20%); and very low for ALV (<5%) antibodies. By commercial ELISA, the seroprevalence of antibodies against AIV was, in some flocks, up to 63%. However, more specific assays did not confirm AIV antibodies, indicating that all flocks tested were free of avian influenza antibodies. Birds seemed to be first infected by IBV (at about 7 wk of age), then by reovirus at 12 wk, before they became infected by APV (week 25) and ILTV (week 30). This is the first report of serological evidence of the above viruses in West Africa. Further studies are necessary to assess economic losses due to these avian viruses and the costs and benefits of countermeasures.


title: Management of upper respiratory tract infections in children.

text: Upper respiratory tract infection (URTI) occurs commonly in both children and adults and is a major cause of mild morbidity. It has a high cost to society, being responsible for absenteeism from school and work and unnecessary medical care, and is occasionally associated with serious sequelae. URTIs are usually caused by several families of virus; these are the rhinovirus, coronavirus, parainfluenza, respiratory syncytial virus (RSV), adenovirus, human metapneumovirus, influenza, enterovirus and the recently discovered bocavirus. This review will mainly focus on the rhinovirus, where significant advances have been made in understanding the epidemiology, natural history and relationship with other pathogens.


title: [Research on coronaviruses in water. I. Adsorption and elution of the coronavirus on glass powder].

text: The purpose of the research was to determine under what conditions enteric coronaviruses from water can be optimally concentrated by acid-pH adsorption on glass-powder followed by alkaline-pH elution. The 14C labelled bovine enteric coronavirus was grown on HRT18 cell monolayers and purified. Virus adsorption was studied in relation to pH: optimal adsorption yield--between 50 to 75% (mean value: 62%)--was obtained at pH less than or equal to 3.5. Elution was made comparatively with a 0.05 M glycine solution and a 3% beef extract solution. Virus elution occurred mostly at alkaline pH with a maximum yield between 38 and 55% for pH values between 9 and 11. Owing to the sensitivity of coronavirus to acid (pH 3) and alkaline pH (pH greater than or equal to 10), adsorption was optimal at pH 3.3 and elution at pH 9. Under such conditions, the overall efficiency of the method appears to stand between 24 and 28%.


title: Fluorescent antibody test for rapid diagnosis of coronaviral enteritis of turkeys (bluecomb).

text: Intestinal tissues obtained from coronavirus-infected embryos and turkeys were examined by fluorescent antibody tissue section technique (FAT). Evidence of viral antigen was demonstrated in the cytoplasm of the intestinal epithelial cells covering the villi. Embryo intestines that were examined from 24 to 96 hours after inoculation were positive for immunofluorescence (IF), whereas bursa of Fabricius was negative. Poults hatched from infected embryos were examined at 2 days of age and were positive for IF. Coronaviral antigen was detected by FAT in the cytoplasm of intestinal epithelial cells of the jejunum, ileum, duodenum, and cecum in all turkeys that were examined from 24 hours to 28 days.


title: Evaluation of a nucleoprotein-based enzyme-linked immunosorbent assay for the detection of antibodies against infectious bronchitis virus.

text: As an immunogen of the coronavirus, the nucleoprotein (N) is a potential antigen for the serological monitoring of infectious bronchitis virus (IBV). In this report, recombinant N protein from the Beaudette strain of IBV was produced and purified from Escherichia coli as well as Sf9 (insect) cells, and used for the coating of enzyme-linked immunosorbent assay (ELISA) plates. The N protein produced in Sf9 cells was phosphorylated whereas N protein from E. coli was not. Our data indicated that N protein purified from E. coli was more sensitive to anti-IBV serum than the protein from Sf9 cells. The recombinant N protein did not react with the antisera to other avian pathogens, implying that it was specific in the recognition of IBV antibodies. In addition, the data from the detection of field samples and IBV strains indicated that using the recombinant protein as coating antigen could achieve an equivalent performance to an ELISA kit based on infected material extracts as a source of antigen(s). ELISAs based on recombinant proteins are safe (no live virus), clean (only virus antigens are present), specific (single proteins can be used) and rapid (to respond to new viral strains and strains that cannot necessarily be easily cultured).


The veterinary profession's duty of care in response to disasters and food animal emergencies.


title: Indomethacin has a potent antiviral activity against SARS coronavirus.

text: UNLABELLED Severe acute respiratory syndrome (SARS) is a newly emerging, highly transmissible and fatal disease caused by a previously unknown coronavirus (SARS-CoV). Existing in non-identified animal reservoirs, SARS-CoV continues to represent a threat to humans because there is no effective specific antiviral therapy for coronavirus infections. OBJECTIVES Starting from the observation that cyclopentenone cyclooxygenase (COX) metabolites are active against several RNA viruses, we investigated the effect of the COX inhibitor indomethacin on coronavirus replication. METHODS Work involving infectious SARS-CoV was performed in biosafety level 3 facilities. SARS-CoV was grown in monkey VERO cells and human lung epithelial A549 cells, while canine coronavirus (CCoV) was grown in A72 canine cells. Antiviral activity was analysed by determining infective virus titres by TCID50, viral RNA synthesis by Northern blot analysis and real-time RT-PCR, and viral protein synthesis by SDS-PAGE analysis after 35S-methionine-labelling. Antiviral efficacy in vivo was determined by evaluating virus titres in CCoV-infected dogs treated orally with 1 mg/kg body weight indomethacin (INDO). RESULTS Unexpectedly, we found that INDO has a potent direct antiviral activity against the coronaviruses SARS-CoV and CCoV. INDO does not affect coronavirus binding or entry into host cells, but acts by blocking viral RNA synthesis at cytoprotective doses. This effect is independent of cyclooxygenase inhibition. INDO's potent antiviral activity (>1,000-fold reduction in virus yield) was confirmed in vivo in CCoV-infected dogs. CONCLUSIONS The results identify INDO as a potent inhibitor of coronavirus replication and suggest that, having both anti-inflammatory and antiviral activity, INDO could be beneficial in SARS therapy.


Bacteraemia caused by Fusobacterium necrophorum: a clinical approach to Lemierre's syndrome.


docid: ocmqatew

title: Cyclodextrins in peptide and protein delivery.

text: The objective of this contribution is to summarize recent findings on the potential of cyclodextrins (CDs) and their derivatives as carriers for therapeutically important peptides, proteins and oligonucleotides. As one of the indices relevant to bioadaptability of CDs in pharmaceutical uses, their interaction with cellular membranes in vitro is outlined. CDs enable the creation of advanced dosage forms for the next generation of drugs that are difficult to formulate and deliver with the existing pharmaceutical excipients. Furthermore, the diagnostic uses of CDs for the direct measurement of cholesterol in high-density and low-density lipoproteins in serum are discussed on the basis of their ability to recognize the surface properties of each lipoprotein particle.


title: Effectiveness of a nosocomial infection control training in improving knowledge in patient-hired attendants and outsourced workers in Taiwan.

text: Nosocomial infection represents an important indicator of healthcare quality and patient safety. Based on the experience gained during the outbreak of Severe Acute Respiratory Syndrome (SARS) in Taiwan in 2003, we gained greater awareness of the potential threat from patient-hired attendants and outsourced workers as bacteria carriers and transmitters. This study built a training model and evaluated training effectiveness in terms of improved knowledge. The training model included nosocomial infection control guidelines, a training CD, training program, evaluation form, and descriptive procedures for conducting training and evaluation. The training course for this study was conducted in 17 hospitals in Taiwan, with a total of 1,467 participants attending 27 seminars and 1,265 effective samples (return rate = 86.2%). Results of the study showed a statistical difference of < .001 before and after the training course on participant understanding of nosocomial infection control. Advantages of applying this training model include knowledge enhancement and reduced implementation time, manpower input and budget outlay. In addition to greater flexibility, the model can also positively influence clinical practice, improve patient safety and reduce hospital nosocomial infection rates. This program is worth promoting in hospitals and other healthcare institutions.


Avoid complacency--prepare for the worst.


title: Japanese Encephalitis Among Three U.S. Travelers Returning from Asia, 2003-2008.

text: Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is a leading cause of encephalitis in Asia. The risk for Japanese encephalitis (JE) for most travelers is low, but varies by travel destination, duration, season, and activities. As part of routine surveillance and diagnostic testing, state health officials or clinicians send specimens from patients with unexplained encephalitis to CDC. To characterize the epidemiologic and clinical features of JE cases, CDC reviewed all laboratory-confirmed cases that occurred during 1992 (when a JE vaccine was first licensed in the United States) to 2008. Four cases were identified, including one previously reported. This report describes the three previously unpublished cases. All were Asian immigrants or family members who traveled to Asia to live or to visit friends or relatives and had not been vaccinated for JE. The three patients experienced fever with mental status changes, but JE was recognized early in the clinical course of only one patient. All recovered, but two patients had residual neurologic deficits. Travelers to Asia might be at increased risk for JE because of rural itineraries and lack of perceived risk. To protect against JE, travelers should seek medical advice on protective measures, including possible JE vaccination, well in advance of departure for Asia. While in Asia, travelers should use personal protective measures to reduce the risk for mosquito bites. Health-care providers should assess the risk for JE in travelers to Asia and provide appropriate preventive or supportive treatment measures.


title: A suture technique for easier reduction and repair of bucket-handle meniscal tears while using the all-inside devices.

text: Arthroscopic repair of bucket-handle meniscal tears is difficult due to their complex pathology. Many meniscal repair techniques such as all-inside, inside-out, and outside-in have been described for the treatment of these tears. Loss of reduction is a likely complication with the use of new-generation, all-inside suture instruments, as the tip of the needle is extracted following advancement of the first implant behind the capsule. The complication may be encountered quite often and renders the use of the meniscus repair instrument unusable and causes an irreparable iatrogenic injury in the meniscus. The application of a simpler and more efficient technique is necessary until surgical experience is gained. The aim of this study was to define a new, simpler, and more efficient combination of suturing method in the treatment of bucket-handle meniscal repairs and minimize the rate of complications which may be caused by this technique.


title: Testing for salmonella spp. In released parrots, wild parrots, and domestic fowl in lowland peru.

text: Wild animal populations face threats from pathogens from both intentionally released captive animals and domestic animals that accompany human settlements. From December 2004 through August 2005, we studied free living macaws and parrots in the Tambopata National Reserve in the Peruvian Amazon and semicaptive domestic fowl in human settlements adjacent to the reserve. In 1992-1993, large macaws (Aras spp.) that were serologically positive for Salmonella Pullorum were released into this reserve, which hosts dense populations of free-living parrots and macaws. We collected cloacal swabs from 64 birds and cultured for Salmonella spp. via standard laboratory methods. All 35 psittacines tested were culture negative for Salmonella spp., while 31% of 29 domestic fowl were culture positive. Our findings suggest that the domestic fowl that accompany human settlement in this region carry and shed Salmonella spp. that could threaten wild bird populations in and around the reserve.


title: Nitric oxide donor drugs: an update on pathophysiology and therapeutic potential.

text: The discovery of the multiple physiological and pathophysiological processes in which nitric oxide (NO) is involved has promoted a great number of pharmacological researches to develop new drugs that are capable of influencing NO production directly and/or indirectly for therapeutic purposes (i.e, NO-releasing drugs, NO-inhibiting drugs, and phosphodiesterase V inhibitors). In particular, the so-called NO donor drugs could actually have an important therapeutic effect in the treatment of many diseases such as arteriopathies (atherosclerosis and its sequelae, arterial hypertension and some forms of male sexual impotence), various acute and chronic inflammatory conditions (colitis, rheumatoid arthritis and tissue remodelling), and several degenerative diseases (Alzheimer's disease and cancer). The old organic nitrates show some well-known pitfalls including the induction of tolerance and acute side effects related to abrupt vasodilation such as cephalea and hypotension, which limit their therapeutic indications. A low therapeutic index (i.e., peroxynitrite toxicity) has always characterised the sydnonimines class. A series of interesting new classes of NO donors are under intense pharmacological investigation and scrutiny (S-nitrosothiols, diazeniumdiolates and NO hybrid drugs), each characterised by a particular pharmacokinetic and pharmacodynamic profile. The most important obstacle in the field of NO donor drugs is represented by the difficulty in targeting NO release, and thereby its effects, to a particular tissue.


title: Recovery of a neurovirulent human coronavirus OC43 from an infectious cDNA clone.

text: This study describes the assembly of a full-length cDNA clone of human coronavirus (HCoV)-OC43 in a bacterial artificial chromosome (BAC). The BAC containing the full-length infectious cDNA (pBAC-OC43(FL)) was assembled using a two-part strategy. The first step consisted in the introduction of each end of the viral genome into the BAC with accessory sequences allowing proper transcription. The second step consisted in the insertion of the whole HCoV-OC43 cDNA genome into the BAC. To produce recombinant viral particles, pBAC-OC43(FL) was transfected into BHK-21 cells. Recombinant virus displayed the same phenotypic properties as the wild-type virus, including infectious virus titers produced in cell culture and neurovirulence in mice.


title: SARS preparedness checklist for state and local health officials.

text: A planning checklist for widespread severe acute respiratory syndrome, modeled on an Association of State and Territorial Health Officials (ASTHO) pandemic influenza planning checklist, was developed jointly by ASTHO, the National Association of County and City Health Officials, and the Centers for Disease Control and Prevention. This checklist, distributed May 2003, has been widely used.


Acute cough in adults.


title: Laparoscopic versus open splenectomy: a single center eleven-year experience.

text: The 11-year experience with open (OS) and laparoscopic (LS) splenectomy at a single center is reported. A total of 201 splenectomies were performed and clinical and demographic data were retrospectively analyzed. Patients were classified according to the type of operation as LS or OS. The mean age of patients was 45.1 +/- 17.1, and 141 patients were male. Out of 43 LS, 40 were done for hematologic causes, and they had a significantly shorter hospital stay compared to OS for hematologic causes (6.87 +/- 2.2 vs. 9.84 +/- 2.9 days; p = 0.000003) and significantly less requirement for blood transfusion (26.2 +/- 93.4 vs. 132.4 +/- 252.3 mL; p = 0.0152). In the OS group, comparison of patients with trauma and those with hematologic causes showed that significantly more males underwent surgery for trauma causes (35 of 43 vs. 16 of 21), hospital stay was longer (18.9 +/- 27.4 vs. 9.8 +/- 2.9 days) and blood requirement higher (708.1 +/- 603.7 mL vs. 132.4 +/- 252.3 mL; p = 0.0004, p = 0.047 and p = 0.000001, respectively). Laparoscopic splenectomy is a safe procedure for spleen removal.


title: Helicases as antiviral drug targets.

text: 1. We have demonstrated for the first time that the helicase of a ribonucleic acid virus, the SARS coronavirus (SARS-CoV), is a valid target for drug development. 2. Using high throughput screen and chemical synthesis, several lead compounds targeting the SARS-CoV helicase have been identified. We have shown that these compounds can inhibit SARS-CoV helicase activity and viral growth in cell culture systems. These compounds can potentially be used to target other viruses.


Telehealth and Outreach in the Post COVID-19 World: The MU Show-Me ECHO Program as a National Model.


title: Through the quarantine looking glass: drug-resistant tuberculosis and public health governance, law, and ethics.

text: The incident in May-June 2007 involving a U.S. citizen traveling internationally while infected with drug-resistant tuberculosis involved the U.S. federal government's application of its quarantine and isolation powers. The incident and the isolation order raised numerous important issues for public health governance, law, and ethics. This article explores many of these issues by examining how the exercise of quarantine powers provides a powerful lens through which to understand how societies respond to and attempt to govern threats posed by dangerous, contagious pathogens. The article considers historical aspects of governmental power to quarantine and isolate individuals and groups; analyzes the current state of quarantine and isolation law in the United States in light of the recent incident with drug-resistant tuberculosis; and explores global aspects of public health governance and law highlighted by this incident.


title: Serum amyloid a is a biomarker of acute exacerbations of chronic obstructive pulmonary disease.

text: RATIONALE Much of the total disease burden and cost of chronic obstructive pulmonary disease (COPD) is associated with acute exacerbations of COPD (AECOPD). Serum amyloid A (SAA) is a novel candidate exacerbation biomarker identified by proteomic screening. OBJECTIVES To assess SAA as a biomarker of AECOPD. METHODS Biomarkers were assessed (1) cross-sectionally (stable vs. AECOPD; 62 individuals) and (2) longitudinally with repeated measures (baseline vs. AECOPD vs. convalescence; 78 episodes in 37 individuals). Event severity was graded (I, ambulatory; II, hospitalized; III, respiratory failure) based on consensus guidelines. MEASUREMENTS AND MAIN RESULTS Presumptively newly acquired pathogens were associated with onset of symptomatic AECOPD. In the cross-sectional study, both SAA and C-reactive protein (CRP) were elevated at AECOPD onset compared with stable disease (SAA median, 7.7 vs. 57.6 mg/L; P < 0.01; CRP median, 4.6 vs. 12.5 mg/L; P < 0.01). Receiver operator characteristics analysis was used to generate area-under-curve values for event severity. SAA discriminated level II/III events (SAA, 0.88; 95% confidence interval, 0.80-0.94 vs. CRP, 0.80; 95% confidence interval, 0.70-0.87; P = 0.05). Combining SAA or CRP with major symptoms (Anthonisen criteria, dyspnea) did not further improve the prediction model for severe episodes. IL-6 and procalcitonin were not informative. CONCLUSIONS SAA is a novel blood biomarker of AECOPD that is more sensitive than CRP alone or in combination with dyspnea. SAA may offer new insights into the pathogenesis of AECOPD.


title: Viral enteritis in calves.

text: A complex community of bacteria, viruses, fungi, protists, and other microorganisms inhabit the gastrointestinal tract of calves and play important roles in gut health and disease. The viral component of the microbiome (the virome) is receiving increasing attention for its role in neonatal calf diarrhea (NCD). Rotavirus and coronavirus have for a long time been associated with NCD and commercial vaccines have been produced against these agents. Recently, several other viruses which may play a role in diarrhea have been discovered in calf fecal samples, mostly by sequence-based methods. These viruses include torovirus, norovirus, nebovirus, astrovirus, kobuvirus, and enterovirus. Most studies have involved epidemiologic investigations seeking to show association with diarrhea for each virus alone or in combination with potential pathogens. However, determining the contribution of these viruses to calf diarrhea has been challenging and much uncertainty remains concerning their roles as primary pathogens, co-infection agents, or commensals.


title: Factors associated with car seat test failure in late preterm infants: A retrospective chart review.

text: BACKGROUND Professional organizations recommend car seat testing of preterm infants before discharge from hospital. Late preterm infants (LPIs, 34(0/7) to 36(6/7) weeks' gestational age) are at the greatest risk for failure, despite often being well. OBJECTIVE To determine the incidence of car seat testing failure in LPIs and associative factors. METHODS A retrospective chart review was performed of inborn LPIs admitted to all levels of newborn care between July 1, 2012 and June 30, 2013. Data collected included maternal demographics, labour and delivery history, and neonatal course. Data were analyzed using backward logistic regression. RESULTS A total of 511 charts were reviewed and 367 LPIs were eligible for inclusion. Of the 313 LPIs tested (mean [± SD] gestational age 36 weeks ±6 days and birth weight 2614±405 g), 80 (26%) failed (95% CI 21% to 31%). Most failed due to desaturations (≥2) of <88% for ≥10 s (n=33 [41%]). Multiple gestation was associated with failure (adjusted OR 2.45 [95% CI 1.44 to 4.18]; P=0.001), and there was a trend toward statistical significance for the variable postnatal age (0.996 [95% CI 0.99 to 1.00]; P=0.05). Infants who passed their car seat test had higher postnatal ages than those who failed (mean difference 39.4 h [95% CI 12.7 h to 66.0 h]; P=0.004). CONCLUSION Twenty-six percent of LPIs failed car seat testing. Ideally, infants should be tested after an appropriate transitional period. The authors identified factors that may be important in designing future, prospective studies in this area. Future research should evaluate the clinical significance of car seat testing and resource utilization.


title: Health risks at the Hajj.

text: Annually, millions of Muslims embark on a religious pilgrimage called the "Hajj" to Mecca in Saudi Arabia. The mass migration during the Hajj is unparalleled in scale, and pilgrims face numerous health hazards. The extreme congestion of people and vehicles during this time amplifies health risks, such as those from infectious diseases, that vary each year. Since the Hajj is dictated by the lunar calendar, which is shorter than the Gregorian calendar, it presents public-health policy planners with a moving target, demanding constant preparedness. We review the communicable and non-communicable hazards that pilgrims face. With the rise in global travel, preventing disease transmission has become paramount to avoid the spread of infectious diseases, including SARS (severe acute respiratory syndrome), avian influenza, and haemorrhagic fever. We examine the response of clinicians, the Saudi Ministry of Health, and Hajj authorities to these unique problems, and list health recommendations for prospective pilgrims.


title: Unilateral laparoscopic ovariectomy in a red fox (Vulpes vulpes) with an ovarian cyst.

text: Unilateral laparoscopic ovariectomy was attempted in a red fox (Vulpes vulpes) with an ovarian cyst through single portal access. The ovarian cyst was resistant to conservative therapy using gonadotropin-releasing hormone. A 10-mm laparoscope with an operating channel was introduced into the abdomen via a 12-mm umbilical portal. The left ovary and cyst (34.1 x 30.8 mm) were fixed to the left abdominal wall by a transabdominal suspension suture. The ovarian pedicles and ligaments were progressively cauterized and transected with a multifunction bipolar electrocoagulation forceps. The resected cystic ovary was exteriorized through the umbilical portal site. The surgical time was 42 min, and no intra- and postoperative complications were encountered. Two months after the surgery mating was observed, and the fox gave birth to three healthy cubs 56 days after the mating. This is the first report of using laparoscopy in the red fox with an ovarian cyst.


COVID-19 Special Column: COVID-19 Hits Native Hawaiian and Pacific Islander Communities the Hardest.


title: Detection of specific antibodies to severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein for serodiagnosis of SARS coronavirus pneumonia.

text: We report the evaluation of recombinant severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) nucleocapsid protein enzyme-linked immunosorbent assay (ELISA)-based antibody tests for serodiagnosis of SARS-CoV pneumonia and compare the sensitivities and specificities of this ELISA for detection of immunoglobulin G (IgG), IgM, IgA, and their combinations with serum samples from 149 healthy blood donors who donated blood 3 years ago as controls and 106 SARS-CoV pneumonia patients in Hong Kong. The specificities of the ELISA for IgG, IgM, and IgA detection were 95.3, 96.6, and 96.6%, respectively, with corresponding sensitivities of 94.3, 59.4, and 60.4%, respectively. The present ELISA appears to be a sensitive test for serodiagnosis of SARS-CoV pneumonia, is much more economical and less labor-intensive than the indirect immunofluorescence assay, and does not require cultivation of SARS-CoV.


title: Isolation of bovine respiratory coronaviruses from feedlot cattle and comparison of their biological and antigenic properties with bovine enteric coronaviruses.

text: OBJECTIVE To isolate bovine coronaviruses from the respiratory tracts of feedlot cattle and compare antigenic and biological properties of these strains with bovine enteric coronaviruses. ANIMALS 5- to 8-month-old mixed-breed cattle at 4 feedlots. PROCEDURE Samples were obtained from the nasal passages for testing. The 13 samples with the highest magnitude of positive values for bovine coronavirus (BCV) were cultured. Ten strains of bovine respiratory coronavirus (BRCV) were adapted successfully to serial passage. After observation of cytopathic effects (CPE) and confirmation of BRCV by immune electron microscopy and immunofluorescence testing, cell culture-adapted strains were cloned by limiting dilution. These isolates then were compared with a panel of bovine enteric coronaviruses (BECV), using hemagglutination (HA), receptor-destroying enzyme activity (RDE), hemagglutination inhibition (HI), and virus neutralization (VN) assays. Antigenic relatedness values then were calculated. RESULTS The BRCV were detected in 105 of 488 (21.5%) of the cattle tested. Of 13 strains tested, 10 were isolated in cell culture. Six of the BRCV strains were similar to 2 strains obtained from neonatal calves with diarrhea and 2 strains from adult cattle with winter dysentery. The other 4 BRCV isolates had high RDE activity against mouse erythrocytes but differed from other strains of BECV Nine of 10 BRCV isolates had properties similar to the 2 BECV subtypes. CONCLUSIONS AND CLINICAL RELEVANCE The BRCV can be isolated from nasal passages of cattle entering feedlots. Most BRCV were similar to BECV strains, although a few had unique properties. Vaccines developed to protect against enteric strains also may protect against respiratory tract strains.


title: Targeted RNA recombination of the membrane and nucleocapsid protein genes between mouse hepatitis virus and bovine coronavirus.

text: The targeted RNA recombination was attempted to substitute the membrane (M) protein gene and part of the nucleocapsid (N) protein gene of mouse hepatitis virus with the corresponding sequences from bovine coronavirus. Using a defective interfering (DI) RNA-like cDNA construct derived from pMH54, 690 nucleotides representing the entire M gene and the 5' most 915 nucleotides of the N gene of the mouse hepatitis virus Albany 4 mutant were attempted to be replaced. Upon infection of cells with Albany 4 followed by transfection with synthetic RNA transcribed from the DI-like cDNA construct, recombinant mouse hepatitis viruses as the large plaque forming phenotype were isolated by plaque assays at the non-permissive temperature of 391 degrees C. By RT-PCR and sequencing, those large plaque phenotypes were confirmed to have contained the thermostable phenotype marker derived from the transfected RNA, demonstrating that recombination occurred between the Albany 4 genomic RNA and the in vitro RNA transcripts. Further analysis of the recombinant viruses indicated that there combination had taken place within the region of 222 nucleotides between positions 916 and 1,137 of the N gene. This is the region immediately downstream of the replacement sequence and the start of the temperature resistant phenotype marker. The results suggest that the M and part of the N genes of bovine coronavirus may not be able to complement the function of those of mouse hepatitis virus. This study redirects our current approach of utilizing the MHV targeted RNA recombination as a means to study bovine coronavirus genetics towards the construction of an infectious cDNA clone.


Letter: A coronavirus-like agent present in faeces of cows with diarrhoea.


title: Persistent infection of neural cell lines by human coronaviruses.

text: Human coronaviruses (HCV) have been associated mainly with infections of the respiratory tract. Accumulating evidence from in vitro and in vivo observations is consistent with the neurotropism of these viruses in humans. To verify the possibility of a persistent infection within the central nervous system (CNS), various human cell lines of neural origin were tested for their ability to maintain chronic infection by both known strains of HCV, OC43 and 229E. Production of infectious progeny virions was monitored by an immunoperoxydase assay on a susceptible cell line and viral RNA was observed after RT-PCR. Astrocytic cell lines U-373 MG and U-87 MG did not sustain a persistent HCV-229E infection, even though they were susceptible to an acute infection by this virus. On the other hand, these two cell lines could maintain a persistent infection by HCV-OC43 for as many as 25 cell passages (about 130 days of culture). Relatively stable titers of infectious viral particles, as well as apparently constant amounts of viral RNA were detected throughout the persistent infection of U-87 MG cells. However, persistent infection of U-373 MG cells was accompanied by the detection of infectious viral particles from passage 0 to passage 13 and then from passage 20 to the end of the experiment. This gap in the production of infectious virions was correlated by a drop in the apparent amount of viral RNA detected at passages 15 and 20. These results confirm the ability of HCV-OC43 to persistently infect cells of an astrocytic lineage and, together with our previous observations of HCV infection of primary cultures of human astrocytes and the detection of HCV RNA in human brains, are consistent with the possibility that this human coronavirus could persist in the human CNS by targeting astrocytes.


title: Risk of respiratory infections in health care workers: lessons on infection control emerge from the SARS outbreak.

text: Close proximity of persons together with handling of human secretions (eg respiratory secretions) make health care workers (HCW) particularly vulnerable to transmission of droplet-transmitted respiratory infections. This was tragically highlighted during the international outbreak of severe acute respiratory syndrome (SARS) in 2003 with attack rates of more than 50% in HCW. The purpose of this article is to review common airborne and droplet-transmitted bacterial and viral respiratory tract infections with regard to their impact on health care workers. Lessons need to be learned from the SARS epidemic. The three main strategies to prevent or control occupationally acquired infections are relatively simple and cost-effective-droplet and contact precautions and for some pathogens also vaccination. Enforced implementation of stringent droplet precautions during the SARS crisis should be maintained; and this will most likely have a major additional impact on other nosocomial infections. Employee health services should proactively and creatively devise delivery systems that enhance compliance with vaccination programs for all health care workers. Hospital surveillance should be expanded to all respiratory diseases to facilitate early detection of nosocomial outbreaks, and this should also include surveillance of all HCW. Integrated syndromic and virological surveillance systems set up during the SARS epidemic will also further our understanding of other respiratory infections in the hospital setting. Even if pursuing early diagnosis for unspecific respiratory illnesses is expensive, identification of the causative organism may reduce unnecessary isolation, contact tracing and anxiety, in particular during an outbreak situation. We have a duty to protect our health care workers.


title: Porcine reproductive and respiratory syndrome virus strains of exceptional diversity in eastern Europe support the definition of new genetic subtypes.

text: Porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 and ORF7 sequences from Belarus were found to be of the European (EU) genotype, but grouped separately from all other EU genotype sequences described so far, including live-attenuated EU genotype PRRSV vaccines and Italian EU genotype sequences, some of which have been associated with reduced vaccine efficacy. Also, the Belarusian EU-PRRSV exhibited extreme ORF7 size polymorphism, ranging from 375 nt (the smallest EU genotype ORF7 yet described) to 393 nt (the largest ORF7 yet described for any arterivirus). With the Belarusian sequences, the diversity of EU genotype PRRSV now exceeds that of the North American (US) genotype PRRSV, suggesting a European origin of PRRSV. Finally, a very sharp geographical demarcation of highly diverse EU genotype PRRSV was observed along the eastern Polish border. The new Belarusian sequences have relevance for vaccine and diagnostic-antigen design and show that sequence analysis of PRRSV from more eastern parts of Europe may offer further insights into the emergence and evolution of PRRSV.


title: Kinetics of in vivo proliferation and death of memory and naive CD8 T cells: parameter estimation based on 5-bromo-2'-deoxyuridine incorporation in spleen, lymph nodes, and bone marrow.

text: To study naive and memory CD8 T cell turnover, we performed BrdU incorporation experiments in adult thymectomized C57BL/6 mice and analyzed data in a mathematical framework. The following aspects were novel: 1) we examined the bone marrow, in addition to spleen and lymph nodes, and took into account the sum of cells contained in the three organs; 2) to describe both BrdU-labeling and -delabeling phase, we designed a general mathematical model, in which cell populations were distinguished based on the number of divisions; 3) to find parameters, we used the experimentally determined numbers of total and BrdU(+) cells and the BrdU-labeling coefficient. We treated mice with BrdU continuously via drinking water for up to 42 days, measured by flow cytometry BrdU incorporation at different times, and calculated the numbers of BrdU(+) naive (CD44(int/low)) and memory (CD44(high)) CD8 T cells. By fitting the model to data, we determined proliferation and death rates of both subsets. Rates were confirmed using independent sets of data, including the numbers of BrdU(+) cells at different times after BrdU withdrawal. We found that both doubling time and half-life of the memory population were approximately 9 wk, whereas for the naive subset the doubling time was almost 1 year and the half-life was roughly 7 wk. Our findings suggest that the higher turnover of memory CD8 T cells as compared with naive CD8 T cells is mostly attributable to a higher proliferation rate. Our results have implications for interpreting physiological and abnormal T cell kinetics in humans.


title: Feline infectious peritonitis virus-infected monocytes internalize viral membrane-bound proteins upon antibody addition.

text: Feline infectious peritonitis virus (FIPV) may cause a highly lethal infection in cats, in spite of a usually strong humoral immune response. Antibodies seem unable to identify infected cells and mediate antibody-dependent cell lysis. In this study, the effect of antibodies on Feline coronavirus (FCoV)-infected monocytes was investigated. Upon addition of FCoV-specific antibodies, surface-expressed viral proteins were internalized through a highly efficient process, resulting in cells without visually detectable viral proteins on their plasma membrane. The internalization was also induced by mAbs against the Spike and Membrane proteins, suggesting that both proteins play a role in the process. The internalization did not occur spontaneously, as it was not observed in cells incubated with medium or non-specific antibodies. Further, the internalization could not be reproduced in feline cell lines, indicating its cell-type specificity. This study sheds new light on the immune-evasive nature of FIPV infections.


title: Epizootic coronaviral typhlocolitis in suckling mice.

text: Multiple epizootics of typhlocolitis associated with high morbidity and mortality occurred among suckling mice in an arbovirology research laboratory. Affected mice had necrosis and hyperplasia of cecal, colonic, and less often, small intestinal mucosa. Epithelial syncytia were present throughout the affected areas. Other organs generally were not involved. The etiologic agent was a coronavirus antigenically related to mouse hepatitis virus strains 1 and S. Sera from dams of affected litters and recovered animals did not contain detectable complement fixing antibody to coronavirus antigen. The lesions and the causative agent differed from previously reported coronaviral syndromes in mice. The source of the infection was not definitely found.


title: The Immunobiology of SARS*.

text: Severe acute respiratory syndrome (SARS) presented as an atypical pneumonia that progressed to acute respiratory distress syndrome in approximately 20% of cases and was associated with a mortality of about 10%. The etiological agent was a novel coronavirus (CoV). Angiotensin-converting enzyme 2 is the functional receptor for SARS-CoV; DC-SIGN and CD209L (L-SIGN) can enhance viral entry. Although the virus infects the lungs, gastrointestinal tract, liver, and kidneys, the disease is limited to the lungs, where diffuse alveolar damage is accompanied by a disproportionately sparse inflammatory infiltrate. Pro-inflammatory cytokines and chemokines, particularly IP-10, IL-8, and MCP-1, are elevated in the lungs and peripheral blood, but there is an unusual lack of an antiviral interferon (IFN) response. The virus is susceptible to exogenous type I IFN but suppresses the induction of IFN. Innate immunity is important for viral clearance in the mouse model. Virus-specific neutralizing antibodies that develop during convalescence prevent reinfection in animal models.


title: Angiotensin-converting enzyme inhibitor captopril attenuates ventilator-induced lung injury in rats.

text: We hypothesized that lung inflammation and parenchymal apoptosis in ventilator-induced lung injury (VILI) are related to ANG II and assessed the ability of the angiotensin-converting enzyme inhibitor captopril to attenuate VILI in rats. Adult male Sprague-Dawley rats were randomized to receive two ventilation strategies for 2 h: 1) tidal volume of 40 ml/kg, respiratory rate of 25 breaths/min, and inspiratory O2 fraction of 0.21 [high-volume, 0 positive end-expiratory pressure (HVZP) group] and 2) injection of captopril (100 mg/kg ip) 30 min before HVZP ventilation (HVZP+CAP group). Another group, which did not receive ventilation, served as the control. Mean arterial pressure was significantly lower in the HVZP+CAP group than in the HVZP group at 2 h of ventilation. Total protein levels were significantly higher in bronchoalveolar lavage fluid (BALF) recovered from HVZP-ventilated rats than from controls. BALF macrophage inflammatory protein-2 and lung ANG II were significantly higher in the HVZP group than in the control and HVZP+CAP groups. Lung ANG II levels correlated positively with BALF protein and macrophage inflammatory protein-2. The number of apoptotic airway and alveolar wall cells was significantly higher in the HVZP and HVZP+CAP groups than in the control group and significantly lower in the HVZP+CAP group than in the HVZP group. These results suggest that the efficiency of captopril to attenuate VILI is related to reduction of inflammatory cytokines and inhibition of apoptosis and indicate that VILI is partly mediated by the local angiotensin system.


title: Involvement of host cellular multivesicular body functions in hepatitis B virus budding.

text: Hepatitis B virus (HBV) is a major human pathogen that chronically infects approximately 350 million people, causing liver disease and liver cancer. HBV virions bud into an endoplasmic reticulum (ER)-associated intracellular compartment, but the mechanisms of HBV assembly, budding, and release remain poorly understood. Budding of retroviruses and some other enveloped RNA viruses from plasma membranes requires host functions involved in protein sorting into late endosomal multivesicular bodies (MVBs). To determine whether budding of DNA-containing HBV virions at intracellular membranes also involves MVB functions, we used immunofluorescence to show that, in human hepatoma cells, HBV envelope protein colocalizes with MVB proteins AIP1/ALIX and VPS4B. We also found that a dominant negative (DN) AIP1 mutant inhibited production and/or release of enveloped virions without significant effects on intracellular nucleocapsid formation, whereas DN VPS4B inhibited both nucleocapsid production and budding. By contrast, DN AIP1 and VPS4 had no effect on the efficiency of release of enveloped, nucleocapsid-lacking HBV subviral particles, which are produced in vast excess over virions, and dramatically increased the release of unenveloped, naked nucleocapsids by an apparently nonlytic route. Thus, host MVB functions are required for efficient budding and release of enveloped HBV virions and may be a valuable target for HBV control. Moreover, HBV enveloped virions, enveloped subviral particles, and unenveloped nucleocapsids are all released by distinct pathways with separate host factor requirements.


title: [Small molecular agents against MERS-CoV infection].

text: Middle East respiratory syndrome coronavirus (MERS-CoV) has caused outbreaks of SARS-like disease with 35% case-fatality rate, mainly in the Middle East. A more severe outbreak of MERS occurred recently in the Republic of Korea, where 186 people contracted the infections, causing great concern worldwide. So far, there has been no clinically available drug for the treatment of MERS-CoV infection. The potential drugs against MERS-CoV mainly consist of monoclonal antibodies, peptides and small molecular agents. Small molecular agents have an advantage of easier synthesis, lower cost in production and relatively higher stability. There is better chance for those candidates to gain a quick development. This article reviews the progress of developing small molecular MERS-CoV agents.


title: Variation in volumes and characteristics of trauma patients admitted to a level one trauma centre during national level 4 lockdown for COVID-19 in New Zealand.

text: AIM The aims of this study were to describe the variation in volumes and types of injuries admitted to a level one trauma centre in New Zealand over two 14-day periods before and during the national level 4 lockdown for COVID-19; and highlight communities at risk of preventable injury that may impact negatively on hospital resources. METHOD A retrospective, descriptive study of prospectively collected data in the Midland Trauma Registry in New Zealand. RESULTS Overall there was a reduction of 43% in all injury-related admissions with significant reductions seen in major injury (50% reduction), males (50% reduction) and children aged 0-14 years (48% reduction). Results for ethnicity and persons aged over 14 years were within 3% deviation of this overall 43% reduction. Injuries at home, particularly falls, predominate. CONCLUSION Despite the significant reduction in admissions during level 4 lockdown, hospitals should continue to provide full services until resource limitations are unavoidable. Immediate messaging is recommended to reduce rates of injury on the farm and at home, specifically falls prevention. Ongoing attention of road users to road safety is essential to reduce the incidence of preventable major injury. These immediate measures can potentially reduce unnecessary pressure on hospital beds and resources during the pandemic.


title: [Development of the cDNA chip for SARS virus and a primary study on the possible molecular mechanism of interferon alpha2b inhibiting the SARS virus replication].

text: BACKGROUND To study the molecular mechanism of interferon alpha2b(IFNalpha2b) inhibiting the SARS virus replication. SARS-associated coronavirus (SARS virus) cDNA chip was developed and applied to detect the virus RNA transcription levels in the interferon-treated and untreated cell cultures, and the mechanism of anti-SARS virus activity of interferon alpha2b in cell culture system was explored. METHODS SARS virus cDNA chip was prepared by comparing the published SARS virus genome sequence, and the cDNA chip was used to study the interferon alpha2b function during SARS virus replication. RESULTS SARS virus cDNA chip was successfully prepared by using PCR method. The results showed that the cDNA chip could be used to detect the viral RNA transcription level. Interferon alpha2b could inhibit almost all the SARS virus gene transcription. An unknown gene at the position 28130-28426 bp, named as U gene, may play an important role during the viral replication. CONCLUSION A SARS virus whole genome cDNA chip was established. It could be used to study the virus molecular biology and antiviral drug screening. The results also showed that interferon alpha2b could inhibit almost the whole virus gene transcription by using the cDNA chip.


title: Renal denervation in the most serious form of resistant arterial hypertension.

text: The aim of our observation was to establish whether or not renal sympathetic denervation (RSD) may help control blood pressure (BP) levels in patients with severe hypertension refractory to pharmacological therapy. Out of a group of 12 patients, candidates for RSD, with uncontrolled hypertension and a systolic BP over 190 mm Hg on repeated measurements despite optimal medication, four patients were excluded for multiple renal arteries and one for hyperaldosteronism. Seven patients had RSD using a Symplicity device (5M, 2 F) with a mean age of 64.9 years. While all were followed up for a minimum of 6 months, follow-up duration in the majority of them was substantially longer (12-20 months). At six months post-RSD, six of the seven patients showed a decrease in systolic BP by at least 15 mm Hg while receiving the same or fewer doses of antihypertensive agents. A similar response was seen in diastolic BP. The BP decrease was maintained throughout whole follow-up. In a small group of patients with severe hypertension, we demonstrated that renal sympathetic denervation is capable of reducing blood pressure even in patients with severe hypertension.


title: Trocar site herniation of the small bowel after laparoscopic bilateral adnexectomy despite fascial closure: a case report.

text: In this paper we report a case of an incarcerated hernia occurring through the peritoneal and muscular defect caused by a previous trocar insertion. The patient developed the hernia eight days after bilateral laparoscopic adnexectomy and presented small bowel obstruction signs. This hernia occurred despite correct closure of the internal oblique fascia. The patient was re-operated and exploratory laparoscopy confirmed the diagnosis. The hernia was reduced via a small extension of the previous incision, and the defect was repaired by separated stitches. This case shows that a trocar site hernia can appear despite correct closure of the fascia, which is poorly described except for obese patients. It suggests the need for careful closure of the abdominal wall including the peritoneum after a laparoscopic procedure. Trocar site hernia has to be considered in cases of post laparoscopic small bowel obstruction. We reviewed the literature and found no randomized control study concerning this problem: only reviews, retrospective studies, case reports and technical notes. These papers are discussed and compared with our case.


docid: uqbertgl

title: Identification of porcine mycoplasmas using commercial enzyme assay systems.


title: Viruses in acute gastroenteritis in children in Pune, India.

text: A 2-year study from January 1981 to December 1982 was undertaken to determine the role of viruses in the causation of diarrhoea in hospitalized children in Pune, Maharashtra State, India. The stool samples of 426 children (213 diarrhoeal and 213 non-diarrhoeal controls) were investigated by electron microscopy and ELISA for the presence of viruses. Six morphologically distinct viruses were visualized: rotavirus, coronavirus-like particles (CVLP), adenovirus, astrovirus, calicivirus and small round virus-like particles (SRV). Rotavirus was detected in 28.6% of the diarrhoeal patients and in 1.4% of the controls. The frequency of infection with rotavirus was highest in the children aged less than 5 years. The mean age of rotavirus-positive patients was 11 months. Although rotavirus was detected in almost every month, there has a seasonal trend for colder months when CVLP cases were fewest. However, the prevalence of CVLP was greater in the control group (23.0%) rather than in those with diarrhoea (8.9%). In the control group, CVLP were detected more frequently during the summer months. An inverse relationship between CVLP and rotavirus was observed in children. Adenovirus, astrovirus, calicivirus and SRV were detected in a small proportion of children with and without clinical symptoms of gastroenteritis.


title: 1H NMR assignment and melting temperature study of cis-syn and trans-syn thymine dimer containing duplexes of d(CGTATTATGC).d(GCATAATACG).

text: The preparation and spectroscopic characterization of duplex decamers containing site-specific cis-syn and trans-syn thymine dimers are described. Three duplex decamers, d(CGTATTATGC).d(GCATAATACG), d(CGTAT[c,s]TATGC).d(GCATAATACG), and d(CGTAT[t,s]TATGC).d(GCATAATACG), were prepared by solid-phase phosphoramidite synthesis utilizing cis-syn and trans-syn cyclobutane thymine dimer building blocks (Taylor et al., 1987; Taylor & Brockie, 1988). NMR spectra (500 MHz 2D 1H and 202 MHz 1D 31P) were obtained in "100%" D2O at 10 degrees C, and 1D exchangeable 1H spectra were obtained in 10% D2O at 10 degrees C. 1H NMR assignments for H5, H6, H8, CH3, H1', H2', and H2" were made on the basis of standard sequential NOE assignment strategies and verified in part by DQF COSY data. Comparison of the chemical shift data suggests that the helix structure is perturbed more to the 3'-side of the cis-syn dimer and more to the 5'-side of the trans-syn dimer. Thermodynamic parameters for the helix in equilibrium coil equilibrium were obtained by two-state, all or none, analysis of the melting behavior of the duplexes. Analysis of the temperature dependence of the T5CH3 1H NMR signal gave delta H = 44 +/- 4 kcal and delta S = 132 +/- 13 eu for the trans-syn duplex. Analysis of the concentration and temperature dependence of UV spectra gave delta H = 64 +/- 6 kcal and delta S = 178 +/- 18 eu for the parent duplex and delta H = 66 +/- 7 kcal and delta S = 189 +/- 19 eu for cis-syn duplex. It was concluded that photodimerization of the dTpdT unit to give the cis-syn product causes little perturbation of the DNA whereas dimerization to give the trans-syn product causes much greater perturbation, possibly in the form of a kink or dislocation at the 5'-side of the dimer.


title: Aetiological role of viral and bacterial infections in acute adult lower respiratory tract infection (LRTI) in primary care.

text: BACKGROUND Lower respiratory tract infections (LRTI) are a common reason for consulting general practitioners (GPs). In most cases the aetiology is unknown, yet most result in an antibiotic prescription. The aetiology of LRTI was investigated in a prospective controlled study. METHODS Eighty adults presenting to GPs with acute LRTI were recruited together with 49 controls over 12 months. Throat swabs, nasal aspirates (patients and controls), and sputum (patients) were obtained and polymerase chain reaction (PCR) and reverse transcriptase polymerase chain reaction (RT-PCR) assays were used to detect Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila, influenza viruses (AH1, AH3 and B), parainfluenza viruses 1-3, coronaviruses, respiratory syncytial virus, adenoviruses, rhinoviruses, and enteroviruses. Standard sputum bacteriology was also performed. Outcome was recorded at a follow up visit. RESULTS Potential pathogens were identified in 55 patients with LRTI (69%) and seven controls (14%; p<0.0001). The identification rate was 63% (viruses) and 26% (bacteria) for patients and 12% (p<0.0001) and 6% (p = 0.013), respectively, for controls. The most common organisms identified in the patients were rhinoviruses (33%), influenza viruses (24%), and Streptococcus pneumoniae (19%) compared with 2% (p<0.001), 6% (p = 0.013), and 4% (p = 0.034), respectively, in controls. Multiple pathogens were identified in 18 of the 80 LRTI patients (22.5%) and in two of the 49 controls (4%; p = 0.011). Atypical organisms were rarely identified. Cases with bacterial aetiology were clinically indistinguishable from those with viral aetiology. CONCLUSION Patients presenting to GPs with acute adult LRTI predominantly have a viral illness which is most commonly caused by rhinoviruses and influenza viruses.


title: Functional characterization of the placental fusogenic membrane protein syncytin.

text: Syncytin is an envelope protein of the human endogenous retrovirus family W (HERV-W). Syncytin is specifically expressed in the human placenta and mediates trophoblast cell fusion into the multinucleated syncytiotrophoblast layer. It is a polypeptide of 538 amino acids and is predicted to be posttranslationally cleaved into a surface (SU) subunit and a transmembrane (TM) subunit. Functional characterization of syncytin protein can aid understanding of the molecular mechanism underlying syncytin-mediated cell fusion. In this report, we studied the structure-function relationship of syncytin in 293T and HeLa cells transiently expressing wild-type syncytin or syncytin mutants generated by linker scanning and deletion mutagenesis. Of the 22 linker-inserted mutants, mutants InS51, InV139, InE156, InS493, InA506, and InL529 were fusogenic, suggesting that regions around amino acids S51, V139, and E156 in the SU subunit and S493, A506, and L529 in the cytoplasmic domain (CTM) of syncytin are flexible in conformation. Of the 17 deletion mutants, nine mutants with deletions in the region from amino acids 479 to 538 were fusogenic. The deletion mutant DelI480, containing only the first four amino acid residues in the cytoplasmic domain, had enhanced fusogenic activity in comparison with the wild-type. In addition, two heptad repeat regions (HRA and B) were defined in the TM subunit of syncytin. A peptide inhibitor derived from the C-terminal heptad repeat region (HRB) was shown to potently inhibit syncytin-mediated cell fusion. Our results suggest that the cytoplasmic domain of syncytin is not essential for syncytin-mediated fusion but may play a regulatory role, and an intramolecular interaction between HRA and B is involved in the fusion process.


docid: bpavqshc

title: COVID-19 is possibly a consequence of the anthropogenic biodiversity crisis and climate changes.