title: National laboratory surveillance of viral agents of gastroenteritis in Brazil.

text: In 1984 an enzyme immunoassay for rotavirus and adenovirus developed in the Virology Department of the Oswaldo Cruz Foundation was distributed to laboratories in 14 Brazilian states as part of a project to survey viral agents in fecal specimens from children with diarrhea. The ensuing surveillance continued for 3 to 5 years, and in 1991 the results obtained by all laboratories that tested 300 or more fecal specimens were reviewed at a workshop held in Belém. These results showed that rotavirus had been detected in 13% to 20% of the specimens tested, with positive specimens appearing to peak in the May to September "winter" or dry season in Brazil's central and southern states but not in the more tropical northern areas. Adenovirus, detected in 0.7% to 5.5% of the specimens tested for it, showed no seasonal variations. Many other viral agents known to cause gastroenteritis (e.g., astrovirus, small round-structured viruses, calcivirus, and group C rotavirus) were detected at centers that used electron microscopy. Picobirnavirus, a novel agent not yet associated with gastroenteritis in humans, was found by polyacrylamide gel electrophoresis in specimens from a number of centers using this technique. Vaccines to prevent rotavirus that are currently under development would be of great use in Brazil, where rotavirus is the most common cause of childhood diarrhea. Improved diagnostics will be required to assess the importance of the other viral agents.


title: Retroperitoneal LESS donor nephrectomy.

text: Donor nephrectomy with laparo-endoscopic single site (LESS) surgery has been reported via the transperitoneal approach. We describe a novel technique of retroperitoneal donor nephrectomy using a single surgical incision in the groin, below the abdominal skin crease or "bikini line". The LESS groin incision offers superior cosmesis, while the retroperitoneal approach has distinct advantages, such as the ability to identify the renal vessels early. The new procedure has been performed in two obese patients (body mass index 32 and 33 kg/m2, respectively). The operative times were 4 and 5 hours, warm ischemic times 135 and 315 seconds, blood loss 100 and 250 mL, and hospitalization 3 and 2 days, respectively. Retroperitoneal LESS donor nephrectomy through a single, inconspicuous groin incision is feasible and safe. Further evaluation of the technique in a larger patient cohort is indicated.


title: Mortality patterns associated with poult enteritis mortality syndrome (PEMS) and coronaviral enteritis in turkey flocks raised in PEMS-affected regions.

text: Poult enteritis mortality syndrome (PEMS) is an economically devastating disease. To date, many questions about the syndrome remain unanswered, including its cause, transmission of causative agent(s), and control methods. Turkey coronavirus (TCV) infection has been associated with some outbreaks of PEMS, with areas having a higher prevalence of TCV infection also experiencing an increased incidence of PEMS. This study was designed to establish mortality patterns for flocks experiencing excess mortality and TCV infection in PEMS-affected regions and to delineate the possible role of TCV in PEMS-affected flocks. Fifty-four commercial turkey flocks on farms in areas with and without a history of TCV infection were monitored for weekly mortality and for antibodies to TCV. Flocks were chosen on the basis of placement dates and were monitored from day of placement until processing. All flocks were tested for TCV by an indirect fluorescent antibody assay. PEMS status was determined with the use of the clinical definition of mortality greater than 2% during any 3-wk period from 2 wk of age through the end of brooding due to unknown cause. Of the 54 flocks, 24 remained healthy, 23 experienced PEMS, and 7 tested positive for TCV but did not experience PEMS. Ten flocks experienced PEMS and tested positive for TCV, whereas 13 flocks experienced PEMS and did not test positive for TCV. Four health status groups were evident: healthy, PEMS positive, TCV positive, and PEMS + TCV positive. Distinct mortality patterns were seen for each of the four health status groups. Whereas TCV was associated with PEMS in 43% of PEMS cases, 13 cases (57%) of PEMS did not involve TCV. Additionally, 7 out of 17 cases of TCV (41%) did not experience excess mortality (PEMS) at any time during brooding of the flock. The results of this study indicate that TCV can be associated with PEMS but is neither necessary nor sufficient to cause PEMS.


title: Antigenic assessment of coronaviruses isolated from patients with multiple sclerosis.

text: Many studies have either supported or discounted the role of coronaviruses as etiologic agents in multiple sclerosis (MS). Two new approaches were applied to investigate this controversy. First, monoclonal antibodies specific for either murine coronaviruses (mouse hepatitis viruses) or human coronaviruses were used to characterize the antigenic features of MS-derived coronaviruses SK and SD. Both isolates were found to have a mouse hepatitis virus-type profile. Second, serum and cerebrospinal fluid antibodies to different coronaviruses, including SD, were measured in MS and control groups. No significant difference in antibody level to coronaviruses was found between MS and control samples. The results of these antigenic studies do not support a specific association between MS and coronaviruses.


title: Laparoscopic cholecystectomy in the treatment of acute cholecystitis: comparison of outcomes and costs between early and delayed cholecystectomy.

text: BACKGROUND Several trials showed that early laparoscopic cholecystectomy is superior to delayed laparoscopic cholecystectomy for the treatment of acute cholecystitis. However actual practice does not conform to current evidence. The aim of this study is to compare outcomes and total hospital costs between early and delayed laparoscopic cholecystectomy for acute cholecystitis. PATIENTS AND METHODS A retrospective analysis of patients with acute cholecystitis that underwent a laparoscopic cholecystectomy at our institutions was performed. Patients were divided into 2 groups on the basis of the treatment received and statistical analysis was performed. RESULTS The study included 91 patients, 52 female and 39 male, with a mean age of 55. Early surgery was performed in 32 cases and delayed surgery in 59 cases. The two groups were comparable for demographics data and severity of disease on admission. There was a no significant difference (p = 0.174) in the mean operative time between early (54.8 min) and delayed group (47.8 min). Conversion rate was higher in the early group (34.3% vs. 20.3%), but difference was not statistically significant (p = 0.223). The overall complications rate was comparable (18.7% early vs. 16.9% delayed, p = 0.941). Length of postoperative stay (4.3 vs. 3.8 days) was similar (p = 0.437), but total hospital stay was significantly 4 days shorter in the early group (p < 0.0001). The mean total cost was higher for the delayed group (4171 vs. 6041), with a significant difference of 1870 Euro (p < 0.0001). CONCLUSIONS Early laparoscopic cholecystectomy has an outcome comparable to the delayed procedure, with a shorter total hospital stay and lower total costs, and it should be considered as the preferred approach in treatment of acute cholecystitis.


title: The proteome of the infectious bronchitis virus Beau-R virion.

text: Infectious bronchitis is a highly contagious respiratory disease of poultry caused by the coronavirus infectious bronchitis virus (IBV). It was thought that coronavirus virions were composed of three major viral structural proteins until investigations of other coronaviruses showed that the virions also include viral non-structural and genus-specific accessory proteins as well as host-cell proteins. To study the proteome of IBV virions, virus was grown in embryonated chicken eggs, purified by sucrose-gradient ultracentrifugation and analysed by mass spectrometry. Analysis of three preparations of purified IBV yielded the three expected structural proteins plus 35 additional virion-associated host proteins. The virion-associated host proteins had a diverse range of functional attributions, being involved in cytoskeleton formation, RNA binding and protein folding pathways. Some of these proteins were unique to this study, while others were found to be orthologous to proteins identified in severe acute respiratory syndrome coronavirus virions and also virions from a number of other RNA and DNA viruses.


title: Importance of the penultimate positive charge in mouse hepatitis coronavirus A59 membrane protein.

text: The coronavirus membrane (M) protein carboxy tail interacts with the nucleocapsid during virus assembly. Previous studies demonstrated that the two terminal residues are important, and the charged residue (R227) in the penultimate position in the mouse hepatitis coronavirus (MHV) A59 M protein was suggested to participate in intermolecular interactions with negative charges in the nucleocapsid (N) protein. To determine the significance of the positive charge at position 227, we substituted the arginine with lysine (K), aspartic acid (D), glutamic acid (E), or alanine (A) and studied these by reverse genetics in the context of a MHV full-length infectious clone. Viruses with wild-type phenotype were readily recovered with the K or A substitutions. In contrast, negative-charge substitutions were not tolerated as well. In all recovered R227D viruses the negative charge was replaced with heterologous residues resulting from apparent template switching during negative-strand synthesis of subgenomic RNA 7. An additional second-site compensatory V202I substitution was present in some viruses. Recovered R227E viruses had second-site changes within the M protein carboxy tail that were partially compensatory. Significantly, most of the second site changes in the R227E mutant viruses were previously shown to compensate for the removal of negative charges in the N protein. Our results strongly indicate that a positive charge is not absolutely required. It is clear that other regions within the tail must also be involved in helping mediate interactions between the M protein and the nucleocapsid.


Preparing for the next flu pandemic: from SARS to avian flu.


title: [Impact of recent advances in molecular techniques on diagnosing lower respiratory tract infections (LRTIs)].

text: At present there is still a great deficit in the routine aetiological diagnosis of lower respiratory tract infections (LRTIs): in most studies more than 50% of cases have no aetiological diagnosis, resulting in prescribing unnecessary or inappropriate antibiotics. A wide variety of diagnostic procedures and techniques are applied to detect the aetiological pathogens of LRTIs. Traditional diagnostic culture methods lack sensitivity, are not feasible in many contexts, and focus only on a few of the large number of aetiological agents. Molecular methods are revolutionizing the diagnostic procedures for managing patients with LRTIs, resulting from a combination of improved sensitivity and specificity, a potential for automatisation and the production of very rapid results.


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title: Identification of a protein linked to the genomic and subgenomic mRNAs of feline calicivirus and its role in translation.

text: 125I protein labelling of oligo(dT)-selected RNA from feline calicivirus (FCV)-infected cells revealed that the genomic and 2.4 kb subgenomic RNAs of FCV are linked to a 15 kDa protein (VPg). Proteinase K treatment of FCV RNA, to remove VPg, led to a decrease in the translatability of the RNA, but there was no obvious change in the site of RNA initiation. Addition of the cap analogue 7-methylGTP to in vitro translations had no effect on the translation of FCV RNA, suggesting that FCV RNA is translated by a cap-independent mechanism. Further evidence that FCV RNA is translated by an unusual mechanism was obtained by translating FCV RNA in vitro at a range of K+ concentrations. FCV RNA was able to direct translation at K+ concentrations at which cellular RNA translation was inhibited.


title: Novel European lineages of bat astroviruses identified in Hungary.

text: KEYWORDS bat, astroviruses, coronaviruses, snRT-PCR, Hungary.


title: Crystal structure of nonstructural protein 10 from the severe acute respiratory syndrome coronavirus reveals a novel fold with two zinc-binding motifs.

text: The severe acute respiratory syndrome coronavirus (SARS-CoV) possesses a large 29.7-kb positive-stranded RNA genome. The first open reading frame encodes replicase polyproteins 1a and 1ab, which are cleaved to generate 16 "nonstructural" proteins, nsp1 to nsp16, involved in viral replication and/or RNA processing. Among these, nsp10 plays a critical role in minus-strand RNA synthesis in a related coronavirus, murine hepatitis virus. Here, we report the crystal structure of SARS-CoV nsp10 at a resolution of 1.8 A as determined by single-wavelength anomalous dispersion using phases derived from hexatantalum dodecabromide. nsp10 is a single domain protein consisting of a pair of antiparallel N-terminal helices stacked against an irregular beta-sheet, a coil-rich C terminus, and two Zn fingers. nsp10 represents a novel fold and is the first structural representative of this family of Zn finger proteins found so far exclusively in coronaviruses. The first Zn finger coordinates a Zn2+ ion in a unique conformation. The second Zn finger, with four cysteines, is a distant member of the "gag-knuckle fold group" of Zn2+-binding domains and appears to maintain the structural integrity of the C-terminal tail. A distinct clustering of basic residues on the protein surface suggests a nucleic acid-binding function. Gel shift assays indicate that in isolation, nsp10 binds single- and double-stranded RNA and DNA with high-micromolar affinity and without obvious sequence specificity. It is possible that nsp10 functions within a larger RNA-binding protein complex. However, its exact role within the replicase complex is still not clear.


title: The effectiveness of acupuncture on pain and mobility in patients with osteoarthritis of the knee: a pilot study.

text: Acupuncture has been repeatedly reported to relieve pain associated with osteoarthritis of the knee. As the vast majority of information on the effectiveness of acupuncture on this condition is based on data collected in Western countries, little is known about patients with osteoarthritis of the knee in Asian countries. In this pilot clinical study, acupuncture was incorporated into the standard care for adult patients with osteoarthritis of the knee to determine its contribution to pain relief and improved mobility. In a prospective, non-randomized controlled study, patients with osteoarthritis of the knee were separated into two groups. The first (the experimental group; n = 12) was scheduled for up to 8 sessions of acupuncture in addition to standard care, while the second (the control group; n = 12) received standard care only. Measurements using the six-minute walking distance test, pain visual analogue scale, and osteoarthritis of the knee outcome measurement were taken at baseline and after 4 weeks. Both study and control groups showed significant improvement with respect to time effects in terms of six-minute walking distance, pain visual analogue scale, pain domain and mobility domain scores determined by the osteoarthritis of the knee outcome measurement (p < .01), after adjusting for covariables. However, improvements measured in the study group did not differ significantly from those in the control group. Patients with osteoarthritis of the knee seemed to experience clinical improvements in six-minute walking distance, pain relief and mobility when their standard care was supplemented with acupuncture.


title: Respiratory tract infections increase susceptibility to neurogenic inflammation in the rat trachea.

text: This study reveals that respiratory tract infections make the tracheal mucosa of rats more susceptible to neurogenic inflammation, which is a type of inflammation mediated by neuropeptides released from sensory nerves. Neurogenic inflammation was produced in the tracheas of 2 groups of Long-Evans rats by electrical stimulation of the vagus nerve (5 V, 1 ms, 20 Hz for 5 min) or by an injection of capsaicin (15 to 200 micrograms/kg i.v.) or substance P (0.05 to 5.0 micrograms/kg i.v.). Rats of one group were pathogen-free; the others had serologic evidence of naturally occurring airway infections caused by Sendai virus, coronavirus, and Mycoplasma pulmonis. The stimuli produced neurogenic inflammation in both groups of rats, but the magnitude of this inflammation was much greater in the infected rats. The susceptibility of the infected rats to neurogenic inflammation was manifested by a 2.0 to 3.1 times larger increase in vascular permeability to Monastral blue, 5 times larger increase in number of neutrophils adhering to the endothelium of venules, and conspicuous morphologic changes in the tracheal epithelium. When pathogen-free rats acquired respiratory tract infections, they too became susceptible to neurogenic inflammation. Other experiments showed that infection by Sendai virus was essential for the change, although infection by M. pulmonis or coronavirus may also be necessary. The susceptibility to neurogenic inflammation outlasted the transient pathologic changes caused in the airway mucosa by the viral infections and may have been permanent.


title: An evaluation of Echinacea angustifolia in experimental rhinovirus infections.

text: BACKGROUND Echinacea has been widely used as an herbal remedy for the common cold, but efficacy studies have produced conflicting results, and there are a variety of echinacea products on the market with different phytochemical compositions. We evaluated the effect of chemically defined extracts from Echinacea angustifolia roots on rhinovirus infection. METHODS Three preparations of echinacea, with distinct phytochemical profiles, were produced by extraction from E. angustifolia roots with supercritical carbon dioxide, 60 percent ethanol, or 20 percent ethanol. A total of 437 volunteers were randomly assigned to receive either prophylaxis (beginning seven days before the virus challenge) or treatment (beginning at the time of the challenge) either with one of these preparations or with placebo. The results for 399 volunteers who were challenged with rhinovirus type 39 and observed in a sequestered setting for five days were included in the data analysis. RESULTS There were no statistically significant effects of the three echinacea extracts on rates of infection or severity of symptoms. Similarly, there were no significant effects of treatment on the volume of nasal secretions, on polymorphonuclear leukocyte or interleukin-8 concentrations in nasal-lavage specimens, or on quantitative-virus titer. CONCLUSIONS The results of this study indicate that extracts of E. angustifolia root, either alone or in combination, do not have clinically significant effects on infection with a rhinovirus or on the clinical illness that results from it.


title: Lymphocyte proliferation responses of pigs inoculated with transmissible gastroenteritis virus or porcine respiratory coronavirus.

text: Cell-mediated immunity was evaluated in intestinal, respiratory, and systemic lymphoid tissues of pigs exposed when 11 days old to virulent transmissible gastroenteritis virus (TGEV), attenuated TGEV, or porcine respiratory coronavirus (PRCV), 3 antigenically related porcine coronaviruses with distinct enteric and respiratory tissue tropisms. Mononuclear cells were prepared from mesenteric lymph nodes (MLN), bronchial lymph nodes (BLN), and spleens of pigs and tested for virus-specific responses by use of lymphocyte proliferation assays. Vigorous MLN and BLN proliferation responses to virulent TGEV and PRCV, respectively, at postinoculation days 8 to 24 were strongly associated with prior detection of TGEV in rectal swab samples and PRCV in nasal swab samples. Gastrointestinal disease and intestinal virus replication, assessed on the basis of rectal virus shedding, were almost exclusively found in the virulent TGEV-inoculated pigs, even though virulent TGEV and a high dose of attenuated TGEV elicited the highest proliferation responses in MLN. Pigs exposed to PRCV or attenuated TGEV did not have clinical signs of disease, and only 1 pig given a high dose of attenuated TGEV shed virus in feces. Porcine respiratory coronavirus replicated in the respiratory tract after either oronasal or aerosol inoculation of virus and induced strong BLN, but not MLN, proliferation responses. A high dose of attenuated TGEV (4 x 10(8) plaque-forming units) was more effective than a lower dose of attenuated TGEV (7 x 10(6) plaque-forming units) in eliciting significant lymphocyte proliferation in MLN and BLN.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Transmissible gastroenteritis coronavirus induces programmed cell death in infected cells through a caspase-dependent pathway.

text: In this report, we show that apoptosis (or programmed cell death) is induced in different cell lines infected with a coronavirus, the porcine transmissible gastroenteritis virus (TGEV). Kinetic analysis of internucleosomal DNA cleavage by agarose gel electrophoresis and flow cytometry or cytometric monitoring of the mitochondrial transmembrane potential showed that, for ST cells infected with TGEV, the first overt signs of apoptosis appeared from 10 to 12 h postinfection on. They preceded morphological changes characteristic of cells undergoing apoptosis, as observed by light and electron microscopy. The tripeptide pan-ICE (caspase) inhibitor N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone blocked TGEV-induced apoptosis with no effect on virus production. The thiol agent pyrrolidine dithiocarbamate inhibited apoptosis, suggesting that TGEV infection may lead to apoptosis via cellular oxidative stress. The effect of TGEV infection on activation of NF-kappaB, a transcription factor known to be activated by oxidative stress, was examined. NF-kappaB DNA binding was shown to be strongly and quickly induced by TGEV infection. However, transcription factor decoy experiments showed that NF-kappaB activation is not critical for TGEV-induced apoptosis.


title: Endoscopic radiofrequency ablation of Barrett's esophagus.

text: Recently, extensive data have been published about the safety and efficacy of endoscopic radiofrequency ablation (RFA) of Barrett's esophagus (BE) with early cancer and dysplasia as well as without dysplasia. RFA has been shown to be effective and safe. Circumferential RFA is delivered using the HALO(360+) Ablation System (Covidien, Inc., Mansfield, MA), which consists ofa high-power energy generator, a sizing balloon catheter, and a number of balloon-based ablation catheters with varying outer diameters. Focal RF energy is delivered using the HALO(90) or HALO(60) Ablation Systems (Covidien, Inc., Mansfield, MA), consisting of a radiofrequency energy generator and an endoscope-mounted electrode. Both RFA systems have official approval to be used in the United States, Europe, and other countries for the treatment of BE as well as in patients with gastric antral vascular ectasia and radiation proctopathy. With increasing widespread use of these systems, a full mastery of the equipment and therapeutic technique is essential to maximize eradication rates of BE while maintaining patient safety. A cost-effective patient selection and eradication protocol for RFA is essential to success with this technique in patients with BE. This article will discuss our experience with RFA treatment of BE using the HALO system for effective eradication of Barrett's dysplasia and early Barrett's cancer and review available data especially from the U.S. National Registry.


Studies with human coronaviruses. II. Some properties of strains 229E and OC43.


title: Ocular and respiratory illness associated with an indoor swimming pool--Nebraska, 2006.

text: On December 26, 2006, the Nebraska Department of Health and Human Services (NDHHS) received a report of a child hospitalized in an intensive care unit for severe chemical epiglottitis and laryngotracheobronchitis after swimming in an indoor motel swimming pool. The pool was inspected the same day and immediately closed by NDHHS because of multiple state health code violations. NDHHS initiated an outbreak investigation to identify additional cases and the cause of the illness. This report describes the results of that investigation, which indicated that 24 persons became ill, and the outbreak likely was the result of exposure to toxic levels of chloramines that had accumulated in the air in the enclosed space above the swimming pool. This outbreak highlights the potential health risks from chemical exposure at improperly maintained pools and the need for properly trained pool operators to maintain water quality.


title: Enzyme-linked immunosorbent assays for the detection of Toxoplasma gondii-specific antibodies and antigens in the aqueous humor of cats.

text: Serum and aqueous humor samples, collected from 14 clinically normal cats and 96 cats with clinical evidence of intraocular inflammation, were assayed with ELISA for Toxoplasma gondii-specific immunoglobulin M (IgM), T gondii-specific IgG, T gondii-specific antigens, total IgG, and total IgM. Additionally, serum was assayed with ELISA for feline leukemia virus p27 antigen and antibodies against the feline immunodeficiency virus as well as with an immunofluorescent antibody assay for antibodies against feline coronaviruses. Calculation of the Goldmann-Witmer coefficient (C-value) for the T gondii-specific antibodies detected in aqueous humor established the likelihood of local antibody production. Serologic evidence of present or prior infection by an infectious agent was found in 81.9% of the clinically affected cats from which serologic results were available (77/94 cats). Seropositive results for toxoplasmosis were found in 74.0% of the clinically affected cats. Anterior segment inflammation was found in 93.1% (81/87 cats from which information was available) of the clinically affected cats, most of which were older males. Toxoplasma gondii-specific antibodies were not detected in the aqueous humor of 6 seropositive, clinically normal cats. The C-values for aqueous T gondii antibodies were greater than 1 in 44.8% of the cats and greater than 8 in 24.0% of the cats. Response to treatment with clindamycin HCl was positive in 15/20 (75%) of the T gondii-seropositive, clinically affected cats treated with this drug. In 13/15 (86.7%) T gondii-seropositive, clinically affected cats having a C-value greater than 1, response to treatment with clindamycin HCl was positive.(ABSTRACT TRUNCATED AT 250 WORDS)


The Netherlands Journal of Medicine's hit list: best cited articles in 2003.


title: Evaluation of an immunofluorescent antibody test to detect bovine herpesvirus 1-specific IgM.

text: An indirect immunofluorescent antibody test (IIFAT) was developed to detect bovine herpesvirus 1 (BHV-1)-specific IgM. All sera were treated with protein-G agarose prior to testing to eliminate the possibility of false-positive results due to IgM-isotype rheumatoid factor (IgM-RF). Specific IgM was first detected 8 days after experimental infection of 3 calves free of maternally derived antibody, with peak responses occurring 2-7 days later. Seroconversion was detected in all 3 calves using a single-dilution enzyme-linked immunosorbent assay. Following reinfection at 30 days postinfection, a low-level IgM response was detected in only 1 calf. Seroconversion was detected in 2 calves. There was no evidence of activation of IgM-RF by infection or reinfection with BHV-1. When 87 acute and convalescent serum pairs collected from 21 outbreaks of respiratory disease were tested, specific IgM was detected in 58 animals (66.6%) from 19 (90.5%) outbreaks. Seroconversion was detected in 44 of these animals (50.6%) from 17 outbreaks (81.0%). The correlations between these 2 assays on a calf and outbreak basis were 79.3% and 90.5%, respectively. Specific IgM was detected in 17/20 sera (85.0%) collected from an additional outbreak. No virus was detected by virus isolation or immunofluorescent staining in nasal mucus samples collected at the same time. Detection of specific IgM by IIFAT is a useful technique for the serodiagnosis of BHV-1 infection.


title: L-SIGN (CD209L) isoforms differently mediate trans-infection of hepatoma cells by hepatitis C virus pseudoparticles.

text: L-SIGN is a C-type lectin that is expressed on liver sinusoidal endothelial cells. Capture of Hepatitis C virus (HCV) by this receptor results in trans-infection of hepatoma cells. L-SIGN alleles have been identified that encode between three and nine tandem repeats of a 23 residue stretch in the juxtamembrane oligomerization domain. Here, it was shown that these repeat-region isoforms are expressed at the surface of mammalian cells and variably bind HCV envelope glycoprotein E2 and HCV pseudoparticles. Differences in binding were reflected in trans-infection efficiency, which was highest for isoform 7 and lowest for isoform 3. These findings provide a molecular mechanism whereby L-SIGN polymorphism could influence the establishment and progression of HCV infection.


title: [RNase III-prepared short interfering RNAs induce degradation of SARS-coronavirus mRNAs in human cells].

text: SARS-associated coronavirus has been identified for the cause of Severe Acute Respiratory Syndrome, for which there is no efficacious drugs or vaccines. RNA interference (RNAi) is a process in cell to degradation specific target mRNA by double-stranded RNA. In mammalian cells, RNAi can be triggered by short interfering RNA (siRNA). RNA interference of virus-specific genes has emerged as a potential antiviral mechanism. This work evaluated if RNase III-prepared short interfering RNAs can induce specific degradation of SARS-coronavirus mRNAs in human cells. Three of SARS genes, RNA dependent RNA polymerase (RdRp), spike and nucleocapsid, were amplified with T7 promoter-flanked primers. Long length double-stranded RNA of these genes were transcribed in vitro and then were cleaved to <30bp length short interfering RNA with E. coli RNase III. These siRNAs were termed esiRNA-R, esiRNA-S and esiRNA-N respectively. RdRp, spike and nucleocapsid DNA fragments were inserted into the plasmid pGL3-Control, obtained plasmids pGL-R, pGL-S and pGL-N can express hybrid mRNAs luciferase-RdRp, spike and -nucleocapsid in cells. Above plasmids and esiRNAs were co-transfected to HEK293F cells with reference plasmid pRL-TK. Firefly luciferase and Renilla luciferase activity were measured. Hybrid mRNAs' abundance was measured using reverse transcription real-time PCR. Firefly luciferase expression of pGL-R was reduced to 13% by esiRNA-R. Expression of pGLS was reduced to 11% by esiRNA-S. Expression of pGL-N was reduced to 40% by esiRNA-N. Control esiRNAs didn't affect luciferase expression; Hybrid mRNAs' abundance was dramatically reduced by corresponding esiRNAs. RNase III-prepared short interfering RNAs induce robust and specific degradation of SARS-coronavirus mRNAs in HEK293F cells. These siRNAs could be used to inhibit SARS-coronavirus in future research.


title: The SANDHOG criteria and its validation for the diagnosis of DCS arising from bounce diving.

text: PURPOSE A three-point scale, the SANDHOG (SAN Diego Diving and Hyperbaric Organizations) criteria, was developed to diagnose DCS (decompression sickness), and then it was validated against a known database of diving related injuries. INTRODUCTION There are currently no universally accepted diagnostic criteria for the diagnosis of DCS. The SANDHOG criteria were developed to address the need for a case definition of DCS. METHODS A point scale and entrance criteria were developed for the diagnosis of DCS. Once the entrance criterion had been met, points were awarded based upon the diver's symptoms and their time of onset. The point system and time limits (SANDHOG criteria) were determined based upon US Navy and Royal Canadian diving reports. The SANDHOG criteria were then applied on a post hoc basis to the Duke Hyperbaric database of diving injuries. Sensitivity and specificity were then calculated using three points as the cut off. The ROC (receiver operating characteristic) analysis was performed to determine the area under the curve (AUC). RESULTS The three point SANDHOG criteria had a specificity of 90.3% and a sensitivity of 52.7%. ROC analysis of the original SANDHOG criteria gave an AUC of 0.72. Using different point values for the diagnosis of DCS will subsequently affect the sensitivity and specificity of the SANDHOG criteria. CONCLUSIONS The specificity of the SANDHOG criteria is good, and demonstrates that the SANDHOG criteria are a useful tool for the diagnosis of DCS.


title: Coronavirus multiplication strategy. II. Mapping the avian infectious bronchitis virus intracellular RNA species to the genome.

text: Avian infectious bronchitis virus, a coronavirus, directed the synthesis of six major single-stranded polyadenylated RNA species in infected chicken embryo kidney cells. These RNAs include the intracellular form of the genome (RNA F) and five smaller RNA species (RNAs A, B, C, D, and E). Species A, B, C, and D are subgenomic RNAs and together with the genome form a nested sequence set, with the sequences of each RNA contained within every larger RNA species (D. F. Stern and S. I. T. Kennedy, J. Virol 34:665-674, 1980). In the present paper we show by RNase T1 oligonucleotide fingerprinting that RNA E is also a member of the nested set. Partial alkaline fragmentation of the genome followed by sucrose fractionation, oligodeoxythymidylate-cellulose chromatography, and RNase T1 fingerprinting gave a partial 3'-to-5' oligonucleotide spot order. A comparison of the oligonucleotides of each of the five subgenomic RNAs with this spot order established that all of the RNAs are comprised of nucleotide sequences inward from the 3' end of the genome. This result is discussed in relation to the multiplication strategy both of coronaviruses and of other RNA-containing viruses.


Acute kidney injury in emerging, non-tropical infections.


title: Structural and functional analysis of an anchorless fibronectin-binding protein FBPS from Gram-positive bacterium Streptococcus suis.

text: The anchorless fibronectin-binding proteins (FnBPs) are a group of important virulence factors for which the structures are not available and the functions are not well defined. In this study we performed comprehensive studies on a prototypic member of this group: the fibronectin-/fibrinogen-binding protein from Streptococcus suis (FBPS). The structures of the N- and C-terminal halves (FBPS-N and FBPS-C), which together cover the full-length protein in sequence, were solved at a resolution of 2.1 and 2.6 Å, respectively, and each was found to be composed of two domains with unique folds. Furthermore, we have elucidated the organization of these domains by small-angle X-ray scattering. We further showed that the fibronectin-binding site is located in FBPS-C and that FBPS promotes the adherence of S suis to host cells by attaching the bacteria via FBPS-N. Finally, we demonstrated that FBPS functions both as an adhesin, promoting S suis attachment to host cells, and as a bacterial factor, activating signaling pathways via β1 integrin receptors to induce chemokine production.


title: SARS in newborns and children.

text: The severe acute respiratory syndrome (SARS) is a highly contagious infection caused by a newly discovered strain of coronavirus (SARS-CoV). Infants born to pregnant women with SARS did not appear to acquire the infection through vertical transmission. Some newborn infants, however, developed severe intrauterine growth retardation and life-threatening gastrointestinal complications. It is now known that the clinical course and prognosis are different between paediatric and adult SARS patients. Young children (< 12 years), in general, run a less aggressive clinical course than do teenage and adult patients. Thus far, no fatalities have been reported in the paediatric age group (< or =18 years). This review describes the current understanding of the clinical manifestations, diagnostic tests, immunological profiles, patient management and outcomes of SARS-CoV infection in the paediatric population.


title: Fatal canine adenoviral pneumonia in two litters of Bulldogs.

text: Five Bulldog pups, 4 weeks of age or younger, were presented over a 2-day period for postmortem examination and diagnostic evaluation. The pups originated from 2 different litters but had been cared for at a common facility since their birth. All 5 pups died after exhibiting symptoms consisting of lethargy, dyspnea, nasal discharge, anorexia, vomiting, diarrhea, and abdominal pain. Necropsy examination revealed locally extensive to diffusely red, firm, consolidated lungs in all pups. Histopathologically, the lungs were variably effaced by multifocal areas of necrosis. The alveolar lumens contained fibrin, edema fluid, macrophages, and neutrophils. Many of the bronchioles contained cellular debris and neutrophils admixed with sloughed bronchiolar epithelium, which often contained large intranuclear amphophilic inclusion bodies that peripherally displaced chromatin. Fluorescent antibody testing was positive for Canine adenovirus. An adenovirus isolated via cell culture was positive on direct fluorescent antibody test and was identified as Canine adenovirus serotype 2 via polymerase chain reaction. Electron microscopy revealed typical viral inclusions within bronchiolar epithelial cells. Hemolytic Escherichia coli was also isolated from the lungs in 3 of the 5 pups. The current case demonstrates a natural and rare fatal infection with a viral agent that is typically associated with immunosuppression in both animals and humans.


title: Management of healthcare waste: developments in Southeast Asia in the twenty-first century.

text: In many Southeast Asian countries, significant challenges persist with regard to the proper management and disposal of healthcare waste. The amount of healthcare waste in these countries is continuously increasing as a result of the expansion of healthcare systems and services. In the past, healthcare waste, if it was treated at all, was mainly incinerated. In the last decade more comprehensive waste management systems were developed for Southeast Asian countries and implementation started. This also included the establishment of alternative healthcare waste treatment systems. The developments in the lower-middle-income countries are of special interest, as major investments are planned. Based upon sample projects, a short overview of the current development trends in the healthcare waste sector in Laos, Indonesia and Vietnam is provided. The projects presented include: (i) Lao Peoples Democratic Republic (development of the national environmental health training system to support the introduction of environmental health standards and improvement of healthcare waste treatment in seven main hospitals by introducing steam-based treatment technologies); (ii) Indonesia (development of a provincial-level healthcare waste-management strategy for Province Nanggroe Aceh Darussalam (NAD) and introduction of an advanced waste treatment system in a tertiary level hospital in Makassar); and (iii) Vietnam (development of a healthcare waste strategy for five provinces in Vietnam and a World Bank-financed project on healthcare waste in Vietnam).


title: SEROLOGICAL SURVEY FOR SELECT INFECTIOUS AGENTS IN WILD MAGELLANIC PENGUINS (SPHENISCUS MAGELLANICUS) IN ARGENTINA, 1994-2008.

text: Despite being the most numerous penguin species in South America, exposure of the Magellanic Penguin (Spheniscus magellanicus) to pathogens has not yet been thoroughly assessed. We collected serum from 1,058 Magellanic Penguins at 10 breeding colonies along the entire latitudinal range of this species in Argentina. The work spanned 10 breeding seasons over 15 yr (1994-2008). Sera were tested for antibodies to select infectious agents. Antibodies reacting against 16 pathogens were detected (seroprevalence): Aspergillus sp. (15.1%), Chlamydia psittaci (6.5%), Salmonella Pullorum (3.1%), Salmonella Typhimurium (81.3%), Aviadenovirus sp. (18.1%), Duck atadenovirus A (23.6%), Anatid herpesvirus 1 (0.7%), Avian orthoreovirus (3.3%), Avian coronavirus M41 (43.5%), Avian coronavirus C46 (59.8%), Avian coronavirus A99 (37.4%), Avian coronavirus JMK (40.2%), Tremovirus A (0.3%), Avian avulavirus 1 (44.0%), Avian avulavirus 2 (43.8%), and Avian avulavirus 3 (46.6%). No antibodies were detected against nine infectious agents: Gallid alphaherpesvirus 1, Gallid alphaherpesvirus 2, Infectious bursal disease virus, Avastrovirus 2, West Nile virus, Eastern equine encephalitis virus, Venezuelan equine encephalitis virus, Western equine encephalitis virus, and Influenza A virus. While restricted by limitations inherent to serological methods, our results provide baseline knowledge for a key species in the South Atlantic Ocean. This information is valuable for adaptive conservation management in a time of increasing environmental stressors affecting the Patagonian Sea, one of the world's richest pelagic seabird communities.


title: An improved procedure for rapid determination of viral RNA sequences of avian RNA viruses.

text: A system for rapid determination of viral RNA sequences, RDV, was improved for detection of avian RNA virus in allantoic fluids. We detected avian paramyxovirus nucleotide sequences using RDV method ver 2.0.


title: Pigs with highly prevalent antibodies to human coronavirus and swine haemagglutinating encephalomyelitis virus in the Tohoku District of Japan.

text: From 1985 to 1988, a total of 2496 swine sera from 60 farms in the Tohoku District of the Honshu Island of Japan were examined for antibodies to swine haemagglutinating encephalomyelitis virus (HEV), human coronavirus (HCV) and bovine coronavirus (BCV) by haemagglutination-inhibition (HI) test. Antibodies to HEV 67N strain and HCV OC43 strain were highly prevalent with positivity rates of 82.1 and 91.4%, respectively, while seropositivity rate to BCV Kakegawa strain was 44.2%. No clinical signs of HEV infection were noticed in any farms including farms with relatively high seropositivity. The results suggested that HCV or antigenitically related virus(es) as well as HEV might be perpetuated in swine in the Tohoku District.


title: [Expression of severe acute respiratory syndrome coronavirus receptors, ACE2 and CD209L in different organ derived microvascular endothelial cells].

text: OBJECTIVE To investigate the expression of the 2 severe acute respiratory syndrome coronavirus (SARS-CoV) receptors, angiotensin-converting enzyme 2 (ACE2) and CD209L in different human organ/tissue derived microvascular endothelial cells. METHODS Endothelial cells from the microvessels in human brain, lung, hepatic sennoside, fat adipose tissue, adrenal gland, esophagus, lymph nodes, and bone were culture. RT-PCR, Western blotting and immunocytochemistry were used to detect the expression of ACE2 and CD209L receptors. RESULTS Both SARS-CoV receptors of ACE2 and CD209L were expressed in the 8 organ/tissue-derived endothelial cells. The expression of ACE2 receptor was the highest in the human lung microvascular endothelial cells, and lowest in the lymphatic endothelial cells. The expression of CD209L was relatively higher in the human lymphatic endothelial cells. CONCLUSION The organ derived microvascular endothelial cells are the important target of SARS-CoV. The pathological injury of lung and lymph system induced by SARS-CoV may be mediated respectively by different receptors of SARS-CoV.


title: Consultation-Liaison Psychiatry Literature Database (2003 update). Part I: Consultation - Liaison Literature Database: 2003 update and national lists.

text: Every day there are 10,000 scientific articles published. Since the Consultation-Liaison ("C-L") psychiatrist may be asked to consult on a patient with any medical illness, e.g., severe acute respiratory syndrome (SARS), malaria, cancer, stroke, amytrophic, lateral sclerosis, and a patient who may be on any medical drug, methods need to be developed to review the recent literature and have an awareness of key and essential current findings. At the same time, teachers need to develop a current listing of seminal papers for trainees and practitioners of this newest cross-over subspecialty of psychiatry-now called Psychosomatic Medicine. Experts selected because of their writings and acknowledged contributions to a specific clinical area or problem hope examined thousands of citations to choose those articles, chapters, books, or letters that they regard as most important to Psychosomatic Medicine. In addition, psychiatric specialists in six countries have provided their national Psychosomatic Medicine (Consultation-Liaison) lists as examples of what they regard as the most important teaching materials journals: Australia, Brazil, Greece, Mexico, Portugal, and Taiwan. It is our belief that a cogent, international, systematic review will provide the greatest success in creating a "regionally appropriate" teaching and consultation literature database with world-wide applicability. We review our current progress on this literature database and software, the technical system and data organization involved, the approach used to populate the literature system, and ongoing development plans to bring this system to the physician via mobile technologies.


Budget delays threaten to leave US science in limbo.


title: Natural A(H1N1)pdm09 influenza virus infection case in a pet ferret in Taiwan.

text: Ferrets have demonstrated high susceptibility to the influenza virus. This study discusses a natural 2009 pandemic influenza A (H1N1) (A(H1N1)pdm09) virus infection in a pet ferret (Mustela putorius furo) identified in Taiwan in 2013. The ferret was in close contact with family members who had recently experienced an influenza-like illness (ILI). The ferret nasal swab showed positive results for influenza A virus using one-step RT-PCR. The virus was isolated and the phylogenetic analysis indicated that all of the eight segmented genes were closely related to the human A(H1N1)pdm09 virus linage isolated in Taiwan. This study may provide a perspective view on natural influenza A virus transmission from the local human population into pet ferrets.


title: SARS: coping with the impact at a community hospital.

text: AIM This paper presents the findings of a staff survey conducted at a 350-bed acute care facility located on the periphery of Toronto, Canada. BACKGROUND Toronto's severe acute respiratory syndrome (SARS) crisis resulted in trauma-like effects at hospitals hardest hit by the disease. A systematic examination of the impact on staff working in hospitals that saw relatively few cases, while maintaining the precautions associated with elevated alert levels, has not been undertaken. METHODS A questionnaire was distributed for 1 month commencing 17 April 2003 and 300 completed responses were obtained (approximately one in six staff members). The data collected included demographic and occupational information, in addition to perceptions of SARS' impact on patient care, factors contributing to adverse impacts on patient care, working conditions, decision-making, communication and relations, sources of support, and the impact on workers' lives outside work. Items for these sections were developed by a multi-disciplinary team of health care workers and hospital administrators. RESULTS In the absence of pre-SARS normative data for the survey, demographic and occupational variables were used to look for patterns of differences between relevant subgroups of respondents. Statistically significant differences were found for gender (73.9% women), nurses (24.7%) vs. others, doctors (20.3%) vs. others, older (40 years or older, 60.0%) vs. younger persons, emergency or intensive care unit workers (8.0%) vs. others, and those employed fewer years at the hospital (less than 5 years: 46.2%) vs. five or more years. These differences varied across the following domains: factors adversely affecting patients, communications, support, working conditions, decision-making and, to a lesser extent, impact on life outside work. While all groups found SARS stressful, nurses reported a greater impact on morale and job satisfaction. Nurses relied more on peer support than doctors, felt less informed and less involved in decision-making than doctors felt, and were more likely to report that infection control procedures were not strict enough. CONCLUSIONS The between-group differences and the pattern of these differences clearly illustrate the polarizing and stressful impact SARS had at a hospital with only a small number of probable or suspect cases. The clear differences between groups defined by demographics, professions and clinical roles suggest a subtle and pervasive secondary impact of the SARS outbreak, with repercussions health care facilities must contend with while maintaining increased levels of vigilance in the wake of SARS.


title: A critical role for the innate immune signaling molecule IRAK-4 in T cell activation.

text: IRAK-4 is a protein kinase that is pivotal in mediating signals for innate immune responses. Here, we report that IRAK-4 signaling is also essential for eliciting adaptive immune responses. Thus, in the absence of IRAK-4, in vivo T cell responses were significantly impaired. Upon T cell receptor stimulation, IRAK-4 is recruited to T cell lipid rafts, where it induces downstream signals, including protein kinase C activation through the association with Zap70. This signaling pathway was found to be required for optimal activation of nuclear factor kappaB. Our findings suggest that T cells use this critical regulator of innate immunity for the development of acquired immunity, suggesting that IRAK-4 may be involved in direct signal cross talk between the two systems.


title: Long-range cooperative interactions modulate dimerization in SARS 3CLpro.

text: Severe acute respiratory syndrome (SARS) is an infectious disease caused by the human coronavirus, SARS-CoV. The main viral protease, SARS 3CLpro, is a validated target for the development of antiviral therapies. Since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. Dimerization inhibitors are usually targeted to the dimerization interface and need to compete with the attractive forces between subunits to be effective. In this paper, we show that the dimerization of SARS 3CLpro is also under allosteric control and that additional and energetically more favorable target sites away from the dimerization interface may also lead to subunit dissociation. We previously identified a cluster of conserved serine residues (Ser139, Ser144, and Ser147) located adjacent to the active site of 3CLpro that could effectively be targeted to inactivate the protease [Bacha, U et al. (2004) Biochemistry 43, 4906-4912]. Mutation of any of these serine residues to alanine had a debilitating effect on the catalytic activity of 3CLpro. In particular, the mutation of Ser147, which does not make any contact with the opposing subunit and is located approximately 9 A away from the dimer interface, totally inhibited dimerization and resulted in a complete loss of enzymatic activity. The finding that residues away from the dimer interface are able to control dimerization defines alternative targets for the design of dimerization inhibitors.


title: Detection of human coronavirus-NL63 in children in Japan.

text: Human coronavirus NL63 recently found in The Netherlands has been detected in Japan with a reverse transcription-polymerase chain reaction technique in clinical specimens from pediatric patients with respiratory symptoms. Of 419 specimens that were negative for common respiratory viruses, 5 were positive for human coronavirus NL63, and these specimens were all collected in the first 3 months of 2003.


Profiling of substrate-specificity and rational design of broad-spectrum peptidomimetic inhibitors for main proteases of coronaviruses.


The role of SARS-CoV protein, ORF-6, in the induction of host cell death.


title: Reovirus apoptosis and virulence are regulated by host cell membrane penetration efficiency.

text: Apoptosis plays an important role in the pathogenesis of reovirus encephalitis and myocarditis in infected animals. Differences in apoptosis efficiency displayed by reovirus strains are linked to the viral mu1-encoding M2 gene segment. Studies using pharmacologic inhibitors of reovirus replication demonstrate that apoptosis induction by reovirus requires viral disassembly in cellular endosomes but not RNA synthesis. Since the mu1 protein functions to pierce endosomal membranes during this temporal window, these findings point to an important role for mu1 in activating signaling pathways that lead to apoptosis. To understand mechanisms used by mu1 to induce apoptosis, a panel of mu1 mutant viruses generated by reverse genetics was analyzed for the capacities to penetrate host cell membranes, activate proapoptotic signaling pathways, evoke cell death, and produce encephalitis in newborn mice. We found that single amino acid changes within the delta region of mu1 reduce the efficiency of membrane penetration. These mutations also diminish the capacities of reovirus to activate proapoptotic transcription factors NF-kappaB and IRF-3 and elicit apoptosis. Additionally, we observed that following intracranial inoculation, an apoptosis-deficient mu1 mutant is less virulent in newborn mice in comparison to the wild-type virus. These results indicate a critical function for the membrane penetration activity of mu1 in evoking prodeath signaling pathways that regulate reovirus pathogenesis.


title: Development of a gonadotrophin-releasing hormone and prostaglandin regimen for the planned breeding of dairy cows.

text: Four studies were carried out to determine the ovarian responses of dairy cows undergoing natural oestrous cycles to sequential injections of gonadotrophin-releasing hormone (GnRH), followed seven days later by prostaglandin and, 48 to 72 hours later, by a second injection of GnRH. In study 1, of 60 cows so treated, 47 were in the intended periovulatory phase when a fixed-time insemination was given 72 hours after the prostaglandin. In study 2, detailed observations were made in 32 cows treated as in study 1, using ultrasound to determine the optimum time to administer the second dose of GnRH. Ovulation was most effectively synchronised by giving GnRH 56 to 60 hours after the prostaglandin. Study 3 investigated the timing of ovulation when no initial dose of GnRH was given. Six cows were injected with prostaglandin on day 12 of the oestrous cycle, followed by GnRH 60 hours later. Five of the six cows ovulated 24 to 36 hours after GnRH, an equivalent timing and synchrony to that in study 2, in which a dose of GnRH had been given seven days before prostaglandin. In study 4, an initial dose of GnRH was given to six cows late (day 17) in the oestrous cycle, and prostaglandin seven days later. The GnRH treatment delayed luteolysis in five of the cows so that they were responsive to the prostaglandin and ovulated 24 to 36 hours after the second dose of GnRH. The use of GnRH (day 0) - prostaglandin (day 7) - GnRH (day 9.5) appears to be an effective means of synchronising ovulation in most cows.


title: Functional domains in the retroviral transmembrane protein.

text: The envelope glycoproteins of the mammalian type C retroviruses consist of two subunits, a surface (SU) protein and a transmembrane (TM) protein. SU binds to the viral receptor and is thought to trigger conformational changes in the associated TM protein that ultimately lead to the fusion of viral and host cell membranes. For Moloney murine leukemia virus (MoMuLV), the envelope protein probably exists as a trimer. We have previously demonstrated that the coexpression of envelope proteins that are individually defective in either the SU or TM subunits can lead to functional complementation (Y. Zhao et al., J. Virol. 71:6967-6972, 1997). We have now extended these studies to investigate the abilities of a panel of fusion-defective TM mutants to complement each other. This analysis identified distinct complementation groups within TM, with implications for interactions between different regions of TM in the fusion process. In viral particles, the C-terminal 16 amino acids of the MoMuLV TM (the R peptide) are cleaved by the viral protease, resulting in an increased fusogenicity of the envelope protein. We have examined the consequences of R peptide cleavage for the different TM fusion mutants and have found that this enhancement of fusogenicity can only occur in cis to certain of the TM mutants. These results suggest that R peptide cleavage enhances the fusogenicity of the envelope protein by influencing the interaction of two distinct regions in the TM ectodomain.


title: Single port VATS resection of a sessile solitary fibrous tumour of the visceral pleura. A case report.

text: UNLABELLED The solitary fibrous tumour of the visceral pleura is a rare neoplasm that derives from mesenchymal cells adjacent to the mesothelial tissue of pleura. Surgical resection is the treatment of choice in benign lesions and healing resulting in half of malignant. Local recurrence can occur in malignant cases, but it is very rare in solitary benign tumors. It may be a result of an incomplete or conservative surgery, lack of identification of a tumor during the operation or a growth of a synchronous neoplasm independent from that removed. Surgical resection is also burdened with some difficulty as the size of the tumor, the relationship with the adjacent structures and identification of the vascular peduncle. We report a case of 72 years-old male with a sessile left solitary fibrous tumour of the visceral pleura. The mass of 10 x 7 x 5 cm was attached, thanks to large planting base, to lateral basal segment of left lower lobe. The patient was treated by single port video assisted approach with a 4 cm skin incision. This case, in our knowledge, represents the first resection by single port VATS of a sessile SFTP with a large plating base and, more generally, the third SFTP resection treated by single portal access. KEY WORDS Solitary fibrous tumour, Uniportal VATS.


title: Engineering the largest RNA virus genome as an infectious bacterial artificial chromosome.

text: The construction of cDNA clones encoding large-size RNA molecules of biological interest, like coronavirus genomes, which are among the largest mature RNA molecules known to biology, has been hampered by the instability of those cDNAs in bacteria. Herein, we show that the application of two strategies, cloning of the cDNAs into a bacterial artificial chromosome and nuclear expression of RNAs that are typically produced within the cytoplasm, is useful for the engineering of large RNA molecules. A cDNA encoding an infectious coronavirus RNA genome has been cloned as a bacterial artificial chromosome. The rescued coronavirus conserved all of the genetic markers introduced throughout the sequence and showed a standard mRNA pattern and the antigenic characteristics expected for the synthetic virus. The cDNA was transcribed within the nucleus, and the RNA translocated to the cytoplasm. Interestingly, the recovered virus had essentially the same sequence as the original one, and no splicing was observed. The cDNA was derived from an attenuated isolate that replicates exclusively in the respiratory tract of swine. During the engineering of the infectious cDNA, the spike gene of the virus was replaced by the spike gene of an enteric isolate. The synthetic virus replicated abundantly in the enteric tract and was fully virulent, demonstrating that the tropism and virulence of the recovered coronavirus can be modified. This demonstration opens up the possibility of employing this infectious cDNA as a vector for vaccine development in human, porcine, canine, and feline species susceptible to group 1 coronaviruses.


title: Murine virus contaminant of Trypanosoma cruzi experimental infection.

text: The possibility that some virus contaminants could be altering host response to Trypanosoma cruzi experimental infection was investigated. Data obtained showed that CBA/J mice infected with stocks of parasite maintained in mice (YIUEC) presented higher level of parasitemia and shorter survival times than those infected with a stock (YITC) which was also maintained in mice but had been previously passaged in cell culture. Mouse antibody production tests, performed with the filtered plasma of mice infected with YIUEC, indicated the presence of mouse hepatitis virus (MHV) while no virus was detected when testing the plasma of YITC infected mice. Filtered plasma of YIEUC infected mice was shown to contain a factor able to enhance the level of parasitemia and to reduce the mean survival time of mice challenged with 10(5) YITC. This factor, that could be serially passaged to naïve mice was shown to be a coronavirus by neutralization tests.


title: CXCR2 signaling and host defense following coronavirus-induced encephalomyelitis.

text: Inoculation of the neurotropic JHM strain of mouse hepatitis virus (JHMV) into the central nervous system (CNS) of susceptible strains of mice results in wide-spread replication within glial cells accompanied by infiltration of virus-specific T lymphocytes that control virus through cytokine secretion and cytolytic activity. Virus persists within white matter tracts of surviving mice resulting in demyelination that is amplified by inflammatory T cells and macrophages. In response to infection, numerous cytokines/chemokines are secreted by resident cells of the CNS and inflammatory leukocytes that participate in both host defense and disease. Among these are the ELR-positive chemokines that are able to signal through CXC chemokine receptors including CXCR2. Early following JHMV infection, ELR-positive chemokines contribute to host defense by attracting CXCR2-expressing cells including polymorphonuclear cells to the CNS that aid in host defense through increasing the permeability the blood-brain-barrier (BBB). During chronic disease, CXCR2 signaling on oligodendroglia protects these cells from apoptosis and restricts the severity of demyelination. This review covers aspects related to host defense and disease in response to JHMV infection and highlights the different roles of CXCR2 signaling in these processes.


title: Coronaviruses' sugar shields as vaccine candidates.

text: A successful global healthcare response relies on versatile vaccines and production of broadly virus-neutralizing antibodies by the immune system to protect us from emerging infectious diseases. The present 2019 severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic highlights the urgent need for development of anti-viral biodefense. Due to the genetic and proteomic diversities of viral pathogens, establishing versatile anti-viral vaccines or therapeutic agents is highly challenging. Carbohydrate antigens represent an important class of immunological targets for vaccine development and immunotherapy against microbial infections. In this mini review, some concepts and strategies for exploring the potential of immunogenic sugar moieties as CoV vaccine candidates are presented.


title: Interaction of hepatitis C virus core protein with viral sense RNA and suppression of its translation.

text: To clarify the binding properties of hepatitis C virus (HCV) core protein and its viral RNA for the encapsidation, morphogenesis, and replication of HCV, the specific interaction of HCV core protein with its genomic RNA synthesized in vitro was examined in an in vivo system. The positive-sense RNA from the 5' end to nucleotide (nt) 2327, which covers the 5' untranslated region (5'UTR) and a part of the coding region of HCV structural proteins, interacted with HCV core protein, while no interaction was observed in the same region of negative-sense RNA and in other regions of viral and antiviral sense RNAs. The internal ribosome entry site (IRES) exists around the 5'UTR of HCV; therefore, the interaction of the core protein with this region of HCV RNA suggests that there is some effect on its cap-independent translation. Cells expressing HCV core protein were transfected with reporter RNAs consisting of nt 1 to 709 of HCV RNA (the 5'UTR of HCV and about two-thirds of the core protein coding regions) followed by a firefly luciferase gene (HCV07Luc RNA). The translation of HCV07Luc RNA was suppressed in cells expressing the core protein, whereas no significant suppression was observed in the case of a reporter RNA possessing the IRES of encephalomyocarditis virus followed by a firefly luciferase. This suppression by the core protein occurred in a dose-dependent manner. The expression of the E1 envelope protein of HCV or beta-galactosidase did not suppress the translation of both HCV and EMCV reporter RNAs. We then examined the regions that are important for suppression of translation by the core protein and found that the region from nt 1 to 344 was enough to exert this suppression. These results suggest that the HCV core protein interacts with viral genomic RNA at a specific region to form nucleocapsids and regulates the expression of HCV by interacting with the 5'UTR.


title: Microvascular exudative hyperresponsiveness in human coronavirus-induced common cold.

text: BACKGROUND The inflammatory response of the airway microcirculation in rhinitis and asthma may be recorded as luminal entry of plasma macromolecules (mucosal exudation). This study examines the exudative responsiveness of the subepithelial microvessels in subjects with and without common cold after inoculation with coronavirus. METHODS The airway mucosa was exposed to exudative concentrations of histamine (40 and 400 micrograms/ml) before and six days after inoculation. To assess whether mucosal penetration of a topically applied agent was altered, nasal absorption of chromium-51 labelled ethylene diamine tetraacetic acid (51Cr-EDTA, MW 372) was also examined. A nasal pool technique kept the challenge and tracer solutes in contact with the same ipsilateral mucosal surface. Concentrations of albumin in lavage fluids were measured as an index of mucosal exudation of plasma. Nasal absorption of 51Cr-EDTA was determined by the cumulated 24 hour urinary excretion of radioactivity. RESULTS Nine subjects developed common cold after coronavirus inoculation and 10 remained healthy. Histamine produced concentration dependent mucosal exudation of plasma in all subjects before and after coronavirus inoculation. In subjects with common cold, however, the histamine-induced mucosal exudation was significantly augmented compared with the group without common cold. This exudative hyperresponsiveness is not explained by an increased baseline exudation because the lavage regimen used produced comparably low baseline exudation in both groups of subjects, nor is it explained by an increased penetration of topical histamine because the ability of the nasal mucosa to absorb 51Cr-EDTA was not significantly increased in the subjects with common cold. CONCLUSIONS An increased proclivity of the airway subepithelial microcirculation to respond with plasma exudation develops during coronavirus-induced common cold. This specific exudative hyperresponsiveness may be a feature of inflammatory airway diseases.


title: Progressive Vascular Functional and Structural Damage in a Bronchopulmonary Dysplasia Model in Preterm Rabbits Exposed to Hyperoxia.

text: Bronchopulmonary dysplasia (BPD) is caused by preterm neonatal lung injury and results in oxygen dependency and pulmonary hypertension. Current clinical management fails to reduce the incidence of BPD, which calls for novel therapies. Fetal rabbits have a lung development that mimics humans and can be used as a translational model to test novel treatment options. In preterm rabbits, exposure to hyperoxia leads to parenchymal changes, yet vascular damage has not been studied in this model. In this study we document the early functional and structural changes of the lung vasculature in preterm rabbits that are induced by hyperoxia after birth. Pulmonary artery Doppler measurements, micro-CT barium angiograms and media thickness of peripheral pulmonary arteries were affected after seven days of hyperoxia when compared to controls. The parenchyma was also affected both at the functional and structural level. Lung function testing showed higher tissue resistance and elastance, with a decreased lung compliance and lung capacity. Histologically hyperoxia leads to fewer and larger alveoli with thicker walls, less developed distal airways and more inflammation than normoxia. In conclusion, we show that the rabbit model develops pulmonary hypertension and developmental lung arrest after preterm lung injury, which parallel the early changes in human BPD. Thus it enables the testing of pharmaceutical agents that target the cardiovascular compartment of the lung for further translation towards the clinic.


title: Nationwide rubella epidemic--Japan, 2013.

text: Rubella usually is a mild, febrile rash illness in children and adults; however, infection early in pregnancy, particularly during the first 16 weeks, can result in miscarriage, stillbirth, or an infant born with birth defects (i.e., congenital rubella syndrome [CRS]). As of 2013, goals to eliminate rubella have been established in two World Health Organization regions (the Region of the Americas by 2010 and the European Region by 2015), and targets for accelerated rubella control and CRS prevention have been established by the Western Pacific Region (WPR). In 1976, Japan introduced single-antigen rubella vaccine in its national immunization program, targeting girls in junior high school. In 1989, a measles-mumps-rubella (MMR) vaccine was introduced, targeting children aged 12-72 months. However, adult males remain susceptible to rubella. From January 1 to May 1, 2013, a total of 5,442 rubella cases were reported through the rubella surveillance system in Japan, with the majority (77%) of cases occurring among adult males. Ten infants with CRS were reported during October 2012-May 1, 2013. Countries and regions establishing a goal of accelerated control or elimination of rubella should review their previous and current immunization policies and strategies to identify and vaccinate susceptible persons and to ensure high population immunity in all cohorts, both male and female.


title: [Evaluation of incidence and risk profile for suffering Covid-19 infection by underlying conditions among middle-aged and older adults in Tarragona.]

text: OBJECTIVE Population-based data on the current Covid-19 pandemic is scarce. This study investigated incidence and risk to suffer Covid-19 by baseline underlying conditions in people ≥50 years in Tarragona region across march-april 2020. METHODS Population-based retrospective cohort study involving 79,071 adults ≥50 years-old in Tarragona region (Southern Catalonia, Spain). Cohort characteristics (age, sex, residence, vaccinations history and comorbidities) were established at baseline, and Covid-19 cases occurring between 01/03/2020-30/04/2020 were registered. Cox regression analysis calculating Hazard ratios (HRs) adjusted by age, sex and comorbidities was used to estimate risk for Covid-19. RESULTS Across study period, 1,547 cohort members were PCR tested (22.6% positive) and 367 were presumptive cases without PCR tested. Considering PCR-confirmed Covid-19, incidence (per 100,000 persons-period) was 441 overall (248, 141, 424, 1,303 and 3,135 in 50-59, 60-69, 70-79, 80-89 and ≥90 years-old, respectively; 380 in men and 497 in women; 259 in community-dwelling and 10,571 in nursing-home). By comorbidities, maximum incidence emerged among persons with neurological disease (2,723), atrial fibrillation (1,348), chronic renal failure (1,050), cardiac disease (856), respiratory disease (798) and diabetes (706). Lower incidence appeared in rheumatic diseases (230) and smokers (180). In multivariable analysis focused on community-dwelling individuals (N=77,671), only cardiac disease (HR: 1.47; 95% CI: 1.01-2.15; p=0.045) and respiratory disease (HR: 1.75; 95% CI: 1.00-3.02; p=0.051) were associated with an increased risk, whereas smoking (HR:0.43; 95% CI: 0.25-0.74; p=0.002) and influenza vaccinated (HR: 0.63; 95% CI: 0.43-0.92; p=0.015) appeared associated with a decreased risk. CONCLUSIONS Apart of increasing age and nursing-home residence, chronic respiratory and cardiac disease appear at increased risk for suffering covid19. This study investigated population-based incidence of Covid-19 infection by underlying conditions among adults ≥50 years in Tarragona (Southern Catalonia, Spain) across two first months pandemic period.


title: Lupus pneumonitis or severe acute respiratory syndrome?

text: Severe acute respiratory syndrome (SARS) is a serious respiratory illness caused by a novel human coronavirus. The disease is highly infectious and carries significant mortality and morbidity. There was a major outbreak of SARS in Guangdong, Taiwan, Beijing, Hong Kong and Toronto between March and June 2003. Common presenting features of SARS are high fever, chills, rigor, malaise, nonproductive cough, lymphopenia and pulmonary infiltrates, followed by rapidly progressive respiratory failure in some cases. We describe two patients with systemic lupus erythematosus (SLE) who presented with fever, systemic upset and pulmonary infiltrates between April and June, 2003. One patient was confirmed to have coronavirus pneumonia while the other had active SLE with lung involvement. Our cases illustrate the difficult diagnostic dilemma in the evaluation of febrile SLE patients during the SARS epidemic.


title: Unintended medication discrepancies at the time of hospital admission.

text: BACKGROUND Prior studies suggest that unintended medication discrepancies that represent errors are common at the time of hospital admission. These errors are particularly worthy of attention because they are not likely to be detected by computerized physician order entry systems. METHODS We prospectively studied patients reporting the use of at least 4 regular prescription medications who were admitted to general internal medicine clinical teaching units. The primary outcome was unintended discrepancies (errors) between the physicians' admission medication orders and a comprehensive medication history obtained through interview. We also evaluated the potential seriousness of these discrepancies. All discrepancies were reviewed with the medical team to determine if they were intentional or unintentional. All unintended discrepancies were rated for their potential to cause patient harm. RESULTS After screening 523 admissions, 151 patients were enrolled based on the inclusion criteria. Eighty-one patients (53.6%; 95% confidence interval, 45.7%-61.6%) had at least 1 unintended discrepancy. The most common error (46.4%) was omission of a regularly used medication. Most (61.4%) of the discrepancies were judged to have no potential to cause serious harm. However, 38.6% of the discrepancies had the potential to cause moderate to severe discomfort or clinical deterioration. CONCLUSIONS Medication errors at the time of hospital admission are common, and some have the potential to cause harm. Better methods of ensuring an accurate medication history at the time of hospital admission are needed.


docid: jmkwgngf

title: [Reflections on translational health research and the COVID-19 case in Argentina].

text: CONICET's Translational Health Research Network is coordinating efforts to advance in translational medicine. Health researchers initiate and focus their research with the aim of improving the health and quality of life of the population. An efficient research system should address health problems relevant to the population resulting in interventions and outcomes important for patients and health professionals. Recommendations to achieve this involve large thematic areas like (a) to set research priorities; (b) to improve research methodology; (c) to make research management and regulation transparent; (d) to increase accessibility to all results; and (e) to improve research dissemination. The recent COVID-19 pandemic has been a clear demonstration of how the country's research system has united the most diverse disciplines to jointly provide solutions to address it. An active and transparent mechanism to identify priorities in the country and to unite funding and research efforts to provide solutions to those priorities is proposed. Translational health research means the joint work of the most diverse health research disciplines in order to jointly obtain efficient and effective interventions to improve the health and quality of life of the population.


title: Ribavirin is not a functional mimic of the 7-methyl guanosine mRNA cap.

text: Ribavirin is a guanosine ribonucleoside analog that displays broad-spectrum anti-viral activity and is currently used for the treatment of some viral infections. Ribavirin has recently been proposed to also be a mimic of the 7-methyl guanosine cap found at the 5' end of mRNAs. To obtain supporting functional data for this hypothesis, we assessed the ability of ribavirin triphosphate to interfere with the interaction between eIF4E and 7-methyl guanosine capped mRNA. In chemical cross-linking assays, cap-affinity chromatography, and cap-dependent translation assays, ribavirin was unable to function as a cap analog.


title: Transmission dynamics of the etiological agent of SARS in Hong Kong: impact of public health interventions.

text: We present an analysis of the first 10 weeks of the severe acute respiratory syndrome (SARS) epidemic in Hong Kong. The epidemic to date has been characterized by two large clusters-initiated by two separate "super-spread" events (SSEs)-and by ongoing community transmission. By fitting a stochastic model to data on 1512 cases, including these clusters, we show that the etiological agent of SARS is moderately transmissible. Excluding SSEs, we estimate that 2.7 secondary infections were generated per case on average at the start of the epidemic, with a substantial contribution from hospital transmission. Transmission rates fell during the epidemic, primarily as a result of reductions in population contact rates and improved hospital infection control, but also because of more rapid hospital attendance by symptomatic individuals. As a result, the epidemic is now in decline, although continued vigilance is necessary for this to be maintained. Restrictions on longer range population movement are shown to be a potentially useful additional control measure in some contexts. We estimate that most currently infected persons are now hospitalized, which highlights the importance of control of nosocomial transmission.


title: Burnout and risk of type 2 diabetes: a prospective study of apparently healthy employed persons.

text: OBJECTIVE This prospective study was designed to test the extent to which the onset of type 2 diabetes in apparently healthy individuals was predicted by burnout, a unique affective response to combined exposure to chronic stressors. METHODS The study participants were 677 employed men and women who were followed up for 3 to 5 years (mean = 3.6 years) for the onset of diagnosed type 2 diabetes. Burnout was assessed by the Shirom-Melamed Burnout Measure with its three subscales: emotional exhaustion, physical fatigue, and cognitive weariness. RESULTS The burnout symptoms were remarkably consistent over the follow-up period irrespective of changes in place of work and in employment status. During the follow-up period, 17 workers developed type 2 diabetes. Logistic regression results indicated that burnout was associated with a 1.84-fold increased risk of diabetes (95% confidence interval [CI] = 1.19-2.85) even after adjusting for age, sex, body mass index, smoking, alcohol use, leisure time physical activity, initial job category, and follow-up duration. In a subsample of 507 workers, the relative risk of diabetes was found to be much higher after additional control for blood pressure levels (odds ratio = 4.32, 95% CI = 1.75-10.67), available only for this subsample. CONCLUSIONS These findings suggest that chronic burnout might be a risk factor for the onset of type 2 diabetes in apparently healthy individuals.


title: Characterization of severe acute respiratory syndrome coronavirus genomes in Taiwan: molecular epidemiology and genome evolution.

text: Since early March 2003, the severe acute respiratory syndrome (SARS) coronavirus (CoV) infection has claimed 346 cases and 37 deaths in Taiwan. The epidemic occurred in two stages. The first stage caused limited familial or hospital infections and lasted from early March to mid-April. All cases had clear contact histories, primarily from Guangdong or Hong Kong. The second stage resulted in a large outbreak in a municipal hospital, and quickly spread to northern and southern Taiwan from late April to mid-June. During this stage, there were some sporadic cases with untraceable contact histories. To investigate the origin and transmission route of SARS-CoV in Taiwan's epidemic, we conducted a systematic viral lineage study by sequencing the entire viral genome from ten SARS patients. SARS-CoV viruses isolated from Taiwan were found closely related to those from Guangdong and Hong Kong. In addition, all cases from the second stage belonged to the same lineage after the municipal hospital outbreak, including the patients without an apparent contact history. Analyses of these full-length sequences showed a positive selection occurring during SARS-CoV virus evolution. The mismatch distribution indicated that SARS viral genomes did not reach equilibrium and suggested a recent introduction of the viruses into human populations. The estimated genome mutation rate was approximately 0.1 per genome, demonstrating possibly one of the lowest rates among known RNA viruses.


title: Clinical Progression of COVID-19 Patient with Extended Incubation Period, Delayed RT-PCR Time-to-positivity, and Potential Role of Chest CT-scan.

text: Coronavirus Disease 2019 (COVID-19), previously called 2019-nCoV, is a novel disease caused by SARS- CoV-2 which was first identified as outbreak of unknown respiratory illness in Wuhan, China. COVID- 19 was declared as global health emergency by WHO on March 11, 2020 and quickly elevated to global pandemic on 11 March 2020. COVID-19 symptom is highly various in each patient, with fever, fatigue, shortness of breath, and cough as the main presenting symptoms. Patient with COVID-19 may shows severe symptom with severe pneumonia and ARDS, mild symptom resembling simple upper respiration tract infection, or even completely asymptomatic. Approximately 80% of cases is mild. However the number may changes as more people are getting tested. Some experts are estimating that up to 50% of all cases may be asymptomatic carrier.


title: Early start of DOAC after ischemic stroke: Risk of intracranial hemorrhage and recurrent events.

text: OBJECTIVE In patients with recent acute ischemic stroke (AIS) and atrial fibrillation, we assessed the starting time of direct, non-vitamin K antagonist oral anticoagulants (DOACs) for secondary prevention, the rate of intracranial hemorrhage (ICH), and recurrent ischemic events during follow-up. METHODS We included consecutive patients with nonvalvular atrial fibrillation admitted to our hospital for AIS or TIA (index event) who received secondary prophylaxis with DOAC or vitamin K antagonists (VKAs). Follow-up was at least 3 months. In the primary analysis, we compared rates of ICH and recurrent ischemic events (AIS or TIA) between patients with early (≤7 days since event; DOACearly) and those with late (>7 days, DOAClate) start of DOAC. RESULTS Two hundred four patients were included (median age 79 years, 89% AIS) and total follow-up time was 78.25 patient-years. One hundred fifty-five patients received DOAC with a median delay of 5 days after the index event (interquartile range 3-11) and 49 received VKA. DOAC was started early in 100 patients (65%). We observed one ICH (1.3%/y) and 6 recurrent AIS (7.7%/y). The ICH occurred in a patient taking VKA. No significant difference in the rate of recurrent AIS between DOACearly (5.1%/y) and DOAClate (9.3%/y, p = 0.53) was observed. CONCLUSIONS Even if DOACs are often started early after an index event, the risk of ICH appears to be low. Among all patients receiving anticoagulation, the rate of recurrent events was 6 times higher than the rate of ICH.


Bright light treatment an effective addition to antidepressants for non-seasonal depression.


title: Histopathology of fasting and bluecomb disease in turkey poults and embryos experimentally infected with bluecomb disease coronavirus.

text: The histopathology of fasting and bluecomb disease in one-day-old turkey poults inoculated with bluecomb disease coronavirus (BCDCV) was studied. Uninoculated fasting poults produced clinical signs similar to those observed in BCDCV-inoculated poults. No histological changes in the intestines were observed in the fasted poults whereas definite lesions were observed in the BCDCV-inoculated poults. The lesions did not differ significantly with whether they were fed or fasted. The severity of the lesions in the intestinal epithelium was in decreasing order in the jejunum, ileum, and cecum. The lesions first appeared 24 hours postinoculation (PI) and progressed through 96 hours PI, as marked shortening of the villi, loss of microvilli, granular appearance of the cytoplasm of epithelial cells with nuclear margination of chromatin, and accentuation of the nucleolus. Similar lesions were observed in the jejunum, ileum, and cecum of turkey embryos inoculated at 24 days old as well as poults from these embryos. Signs of healing were first seen at 120 hours PI. No histopathological changes were observed in the pancreas, brain, kidneys, liver, adrenal, and bursa of Fabricius. The intestinal lesions observed should be a useful histological technique for differentiating fasting from bluecomb disease in turkey poults.


title: Factors controlling coronavirus infections and disease of the central nervous system. A review.

text: There is a correlation between specificity of tropism of JHMV for O-2A lineage cells from the rat and demyelination of white matter, associated with chronic disease. Susceptibility to infection, which can occur in O-2A cells before terminal differentiation may be influenced by cytokines. During the normal, age-related or rapidly induced maturation/differentiation of rat oligodendrocytes, suppression of JHMV replication is correlated with upregulation of the subunit R1 of the cAMP-dependent protein kinase. Virus inhibition occurs at a stage between penetration and initiation of genome expression. Regulation over coronavirus infection of oligodendroglia is strictly controlled by the host cell. There is evidence that induction of R1 subunit of protein kinase A influences uncoating, illustrated in Figure 9, by suppression of dephosphorylation during penetration. Our former working hypothesis, now borne out by recent data predicts that the infection in mature oligodendrocytes is blocked because specific dephosphorylation of the capsid protein N, required for uncoating, etc., is suppressed.


title: Genomic RNA of the murine coronavirus JHM.

text: Genomic RNA extracted from the purified murine coronavirus JHM sediments between 52S and 54S in aqueous sucrose gradients. The RNA is single-stranded and has an apparent mol. wt. of 5.4 to 6.5 X 10(6), as determined by electrophoresis in polyacrylamide agarose gels of different concentrations. The presence of polyadenylate sequences in the RNA is demonstrated by binding to oligo-)dT) cellulose and digestion with ribonucleases A and T1. The purified RNA does not dissociate into subunits at high temperatures or in high concentrations of DMSO and is infectious.


title: Coronavirus antibody detection in cats by computer-assisted kinetics-based enzyme-linked immunosorbent assay (KELA): field studies.

text: A total of 2238 feline serum samples submitted to the New York State Diagnostic Laboratory over a 1-year period were tested for the presence of coronavirus antibodies, using the computer-assisted, kinetics-based enzyme-linked immunosorbent assay (KELA). Cats from which sera were obtained were categorized by sex, age, breed, and disease status, and variations in mean antibody titers for different sub-classifications within each category were analyzed by computerized statistical analysis. As expected, higher mean antibody titers were recorded for cats with feline infectious peritonitis, and for cats with a recent history of possible coronavirus exposure. However, an unexpected inverse relationship between coronavirus antibody titer and age was also found. Certain cattery-oriented pure breeds appeared to have higher mean antibody titers, because their sample populations contained a higher percentage of younger cats and cats of unknown age-groups which, over-all, had higher mean titers. Taken together, the data substantiated the efficacy of the computer-assisted KELA for routine detection of serum coronavirus antibodies in cats.


title: Competition ELISA, using monoclonal antibodies to the transmissible gastroenteritis virus (TGEV) S protein, for serologic differentiation of pigs infected with TGEV or porcine respiratory coronavirus.

text: Monoclonal antibodies (MAB) to subsite A (25C9) and subsite D (44C11) of the S protein of transmissible gastroenteritis virus (TGEV) were used in a blocking ELISA on fixed TGEV-infected swine testis cells to differentiate sera from pigs experimentally inoculated with either TGEV or porcine respiratory coronavirus (PRCV). Serum samples were obtained from pigs at various intervals from postinoculation day (PID) 0 through at least PID 22 to 40. Eleven-day-old pigs, seronegative for TGEV-neutralizing antibodies at the time of inoculation, were inoculated orally and nasally with either the virulent Miller (M5C) strain or the attenuated Purdue (P115) strain of TGEV, or with the ISU-1 strain of PRCV. Gastroenteritis was observed in 100% of the M5C-TGEV-inoculated pigs; but clinical signs of disease were not observed in either the P115-TGEV- or PRCV-inoculated pigs. Virus-neutralization (VN) antibody titer in sera was determined by use of a plaque-reduction assay. Blocking ELISA antibody titer for subsites A and D was determined from the serum dilution that produced 50% reduction in the absorbance values when it competed with biotinylated MAB 25C9 and 44C11, respectively. In sera from the inoculated pigs, the VN antibody titer began to increase by PID 7 and reached maximum by PID 15 to 16. For pigs inoculated with TGEV M5C, subsite A and subsite D blocking antibody titers in the serum paralleled the VN antibody titer, began to increase after PID 7, and reached maximum by PID 15 to 16.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Redirecting coronavirus to a nonnative receptor through a virus-encoded targeting adapter.

text: Murine hepatitis coronavirus (MHV)-A59 infection depends on the interaction of its spike (S) protein with the cellular receptor mCEACAM1a present on murine cells. Human cells lack this receptor and are therefore not susceptible to MHV. Specific alleviation of the tropism barrier by redirecting MHV to a tumor-specific receptor could lead to a virus with appealing properties for tumor therapy. To demonstrate that MHV can be retargeted to a nonnative receptor on human cells, we produced bispecific adapter proteins composed of the N-terminal D1 domain of mCEACAM1a linked to a short targeting peptide, the six-amino-acid His tag. Preincubation of MHV with the adapter proteins and subsequent inoculation of human cells expressing an artificial His receptor resulted in infection of these otherwise nonsusceptible cells and led to subsequent production of progeny virus. To generate a self-targeted virus able to establish multiround infection of the target cells, we subsequently incorporated the gene encoding the bispecific adapter protein as an additional expression cassette into the MHV genome through targeted RNA recombination. When inoculated onto murine LR7 cells, the resulting recombinant virus indeed expressed the adapter protein. Furthermore, inoculation of human target cells with the virus resulted in a His receptor-specific infection that was multiround. Extensive cell-cell fusion and rapid cell killing of infected target cells was observed. Our results show that MHV can be genetically redirected via adapters composed of the S protein binding part of mCEACAM1a and a targeting peptide recognizing a nonnative receptor expressed on human cells, consequently leading to rapid cell death. The results provide interesting leads for further investigations of the use of coronaviruses as antitumor agents.


docid: drltidre

title: Detection of monokines in paraffin-embedded tissues of pigs using polyclonal antibodies.

text: Monokines are glycoproteins, synthesised by macrophages, which exert various effects on the organism. The most important monokines are interleukin (IL)-1alpha, IL-1beta, tumor necrosis factor (TNF)-alpha and IL-6. This paper reports on immunohistochemical techniques developed for the detection of IL-alpha, IL-1beta, IL-6 and TNF-alpha in fixed and paraffin-embedded pig tissues (spleen, lymph nodes, thymus, liver and kidney). Different fixatives (buffered formalin, acetic formalin, paraformadehyde-lysine-periodate and Bouin solution), and antigen unmasking techniques (permeabilisation with Tween 20, pronase enzymatic digestion and microwave-citrate buffer) were used. We describe different protocols for detection of monokines using polyclonal antibodies against the studied monokines. No signal was obtained with monoclonal antibodies against pig-TNF-alpha and human IL-1alpha. Bouin solution was shown to be the best fixative for immunohistochemical detection of IL-1alpha, TNF-alpha, and IL-6, using permeabilisation with Tween 20 as an unmasking antigen method. Acetic formalin was shown to be the best fixative for IL-1beta detection, not needing antigen retrieval techniques. Macrophages were identified as the main cytokine-producing cells, although other types of cells also stained positively to some cytokines. These techniques represent valuable tools for studies of the pathogenesis of viral and bacterial diseases, and of the immune system of the pigs.


title: Successful Conservative Management of Tracheal Injury After Forceful Coughing During Extubation: A Case Report.

text: A-56-year-old woman underwent carpal tunnel release surgery under general anesthesia. Thirty minutes after extubation, the patient complained of chest discomfort with dyspnea. Swelling of the neck and upper anterior chest was observed. Computed tomography of the chest showed tracheal rupture at the brachiocephalic trunk level, and bronchoscopy demonstrated a 5 cm linear tracheal defect in the posterior membranous wall, 6 cm proximal to the carina. Surgical repair of the tracheal injury was impossible due to its location. The patient was managed with intubation, mechanical ventilator care, and antibiotics. She made a full and uncomplicated recovery and was discharged 18 days after the original injury. When suspicious symptoms appear in patients receiving mechanical ventilation support, an immediate and accurate diagnostic process should be undertaken to rule out endotracheal tube-related tracheal injuries and to avoid potentially lethal complications.


title: Genetic diversity of equine arteritis virus.

text: Equine arteritis viruses (EAV) from Europe and America were compared by phylogenetic analysis of 43 isolates obtained over four decades. An additional 22 virus sequences were retrieved from GenBank. Fragments of the glycoprotein G(L) and the replicase genes were amplified by RT-PCR, prior to sequencing and construction of phylogenetic trees. The trees revealed many distinctive lineages, consistent with prolonged diversification within geographically separated host populations. Two large groups and five subgroups were distinguished. Group I consisted mainly of viruses from North America, whilst group II consisted mainly of European isolates. In most instances, where the geographic origin of the viruses appeared to be at variance with the phylogenetically predicted relationships, the horses from which the viruses were recovered had been transported between Europe and America or vice versa. Analysis of the replicase gene revealed similar phylogenetic relationships although not all of the groups were as clearly defined. Virus strains CH1 (Switzerland, 1964) and S1 (Sweden, 1989) represented separate 'outgroups' based on analysis of both genomic regions. The results of this study confirm the value of the G(L) gene of EAV for estimating virus genetic diversity and as a useful tool for tracing routes by which EAV is spread. In addition, computer-assisted predictions of antigenic sites on the G(L) protein revealed considerable variability among the isolates, especially with respect to regions associated with neutralization domains.