title: Attenuation, safety, and efficacy of an infectious bronchitis virus GA98 serotype vaccine.

text: In 1998, novel strains of infectious bronchitis virus (IBV) were identified in chickens from the southeastern United States and classified as a new serotype designated Georgia 98 (GA98). Because of the widespread nature of the GA98 virus in the southeastern United States and the lack of adequate protection with the DE072 vaccine, we developed a specific vaccine for the GA98 serotype. The GA98/0470/98 isolate of IBV was passaged in embryonating chicken eggs 70 times, and attenuation of the virus was determined in specific-pathogen-free chicks. Pass 70 of the GA98/0470/98 strain of IBV when given at 1 day of age by coarse spray and at 14 days of age in the drinking water at 1 x 10(4.5) 50% embryo infectious dose/bird protected against the homologous GA98 challenge as well as provided good protection against the DE072-type virus. In addition, the vaccine was shown to be adequately attenuated and safe at a 10x dosage.


title: Thrombocytopenia in patients with severe acute respiratory syndrome (review).

text: Severe Acute Respiratory Syndrome (SARS) has been recognized as a new human infectious disease caused by a novel coronavirus (SARS-CoV). Hematological changes in patients with SARS were common, including notably lymphopenia and thrombocytopenia. While the former is the result of decreases in CD4+ or CD8+ T-lymphocytes related to the onset of disease or use of glucocorticoids, the latter may involve a number of potential mechanisms. Although the development of autoimmune antibodies or immune complexes triggered by viral infection may play a significant role in inducing thrombocytopenia, SARS-CoV may also directly infect hematopoietic stem/progenitor cells, megakaryocytes and platelets inducing their growth inhibition and apoptosis. Moreover, the increased consumption of platelets and/or the decreased production of platelets in the damaged lungs are a potential alternative mechanism that can contribute to thrombocytopenia in severe critical pulmonary conditions, which has been rarely revealed and will be discussed.


docid: yjtxtjsh

title: Ligasure hemorrhoidectomy versus Stapled hemorrhoidopexy: a prospective randomized clinical trial.

text: AIM The aim of this study was to compare the outcome of Ligasure hemorrhoidectomy (LH) and Stapled hemorrhoidopexy (SH) for prolapsed hemorrhoids. METHODS Sixty-eight patients with grades III and IV hemorrhoids were randomized into two groups of 34 each, to undergo LH (Group 1) or SH (Group 2). Patient demographics, operative details, postoperative pain score, number of parenteral analgesic injections, hospital stay, and time to return to work were all prospectively collected. Postoperative complications and recurrence of prolapse were also recorded. All patients were regularly followed-up for a total period of 18 months. RESULTS Patient demographics and clinical characteristics were similar between both groups. The mean operating time, postoperative pain score, parenteral analgesics, hospital stay and time off work were not statistically significant between both groups. Likewise, both groups had similar postoperative complications except for a residual prolapse that was observed, at four weeks postoperatively, in eight patients (23.53%) in the SH group as compared to two patients (5.89%) in the LH group (P=0.040). Although hemorrhoid recurrence, at 18 months, was also higher among the SH group as compared to the LH group (11.76% vs. 2.94%, respectively), yet the difference was not statistically significant (P=0.163). CONCLUSION Both LH and SH yield comparable good results and minimal side effects for the treatment of grades III and IV hemorrhoids, with less residual prolapses observed with LH. Further, owing to their low postoperative pain, short hospital stay and rapid return to work, both procedures offer an excellent therapeutic option for prolapsed grades III and IV hemorrhoids.


title: Coronavirus proteins: structure and function of the oligosaccharides of the avian infectious bronchitis virus glycoproteins.

text: The recent finding that the E1 glycoproteins of murine coronaviruses contain only O-linked oligosaccharides suggested that this unusual modification might be a distinguishing feature of coronaviruses and might play an essential role in the life cycle of this family of viruses. To examine these possibilities, we analyzed the oligosaccharide moieties of the membrane proteins of the avian coronavirus infectious bronchitis virus. In addition, we determined the effect of inhibiting the glycosylation of these proteins on viral maturation and infectivity. Infectious bronchitis virus virions contain nine proteins. Four of these proteins, GP36, GP31, GP28, and P23, are closely related structurally and appear to be homologous to the E1 proteins of murine coronaviruses. We found that the oligosaccharides of GP31 and GP28 could be removed with endoglycosidase H and that neither of these glycoproteins was detectable in tunicamycin-treated cells. These two results indicated that GP31 and GP28 contain N-linked oligosaccharides. Therefore, O-linked oligosaccharides are not a universal feature of the small coronavirus membrane glycoproteins. Tunicamycin inhibited glycosylation of all of the viral glycoproteins but did not inhibit production of virions by infectious bronchitis virus-infected cells. The virions released by these cells contained only the three non-glycosylated viral proteins P51, P23, and P14. These particles were not infectious. Therefore, it appears that glycosylated infectious bronchitis virus polypeptides are not required for particle formation. However, the viral glycoproteins are apparently indispensible for viral infectivity.


title: Public health systems research: setting a national agenda.

text: The Institute of Medicine has recommended that policy decisions about improvement of national public health systems be guided by sound scientific evidence. However, to date there is no national research agenda to help guide public health systems. The Centers for Disease Control and Prevention was called upon to lead a collaborative consensus-based process to define key research questions and establish a framework to create opportunities to better coordinate, leverage, and identify public health resources, which are increasingly scarce. The public health systems research agenda that emerged from this process has 14 over-arching priority research themes. This national agenda should stimulate and guide research to meet the urgent need to improve the nation's public health systems.


title: Toward prediction of class II mouse major histocompatibility complex peptide binding affinity: in silico bioinformatic evaluation using partial least squares, a robust multivariate statistical technique.

text: The accurate identification of T-cell epitopes remains a principal goal of bioinformatics within immunology. As the immunogenicity of peptide epitopes is dependent on their binding to major histocompatibility complex (MHC) molecules, the prediction of binding affinity is a prerequisite to the reliable prediction of epitopes. The iterative self-consistent (ISC) partial-least-squares (PLS)-based additive method is a recently developed bioinformatic approach for predicting class II peptide-MHC binding affinity. The ISC-PLS method overcomes many of the conceptual difficulties inherent in the prediction of class II peptide-MHC affinity, such as the binding of a mixed population of peptide lengths due to the open-ended class II binding site. The method has applications in both the accurate prediction of class II epitopes and the manipulation of affinity for heteroclitic and competitor peptides. The method is applied here to six class II mouse alleles (I-Ab, I-Ad, I-Ak, I-As, I-Ed, and I-Ek) and included peptides up to 25 amino acids in length. A series of regression equations highlighting the quantitative contributions of individual amino acids at each peptide position was established. The initial model for each allele exhibited only moderate predictivity. Once the set of selected peptide subsequences had converged, the final models exhibited a satisfactory predictive power. Convergence was reached between the 4th and 17th iterations, and the leave-one-out cross-validation statistical terms--q2, SEP, and NC--ranged between 0.732 and 0.925, 0.418 and 0.816, and 1 and 6, respectively. The non-cross-validated statistical terms r2 and SEE ranged between 0.98 and 0.995 and 0.089 and 0.180, respectively. The peptides used in this study are available from the AntiJen database (http://www.jenner.ac.uk/AntiJen). The PLS method is available commercially in the SYBYL molecular modeling software package. The resulting models, which can be used for accurate T-cell epitope prediction, will be made freely available online (http://www.jenner.ac.uk/MHCPred).


Analysis of the cis-acting elements of coronavirus transcription.


Natural killer (NK) cell activity against enteric murine coronavirus mediated by intestinal leukocytes.


title: Evaluation of ProMED-mail as an electronic early warning system for emerging animal diseases: 1996 to 2004.

text: OBJECTIVE To identify emerging animal and zoonotic diseases and associated geographic distribution, disease agents, animal hosts, and seasonality of reporting in the Program for Monitoring Emerging Diseases (ProMED)-mail electronic early warning system. DESIGN Retrospective study. SAMPLE POPULATION 10,490 disease reports. PROCEDURES Descriptive statistics were collated for all animal disease reports appearing on the ProMED-mail system from January 1, 1996, to December 31, 2004. RESULTS Approximately 30% of reports concerned events in the United States; reports were next most common in the United Kingdom, Canada, Australia, Russia, and China. Rabies, bovine spongiform encephalopathy, and anthrax were reported consistently over the study period, whereas avian influenza, Ebola virus, and Hantavirus infection were reported frequently in approximately half of the study years. Reports concerning viral agents composed more than half of the postings. Humans affected by zoonotic disease accounted for a third of the subjects. Cattle were affected in 1,080 reports, and wildlife species were affected in 825 reports. For the 10,490 postings studied, there was a retraction rate of 0.01 and a correction rate of 0.02. CONCLUSIONS AND CLINICAL RELEVANCE ProMED-mail provided global coverage, but gaps in coverage for individual countries were detected. The value of a global electronic reporting system for monitoring emerging diseases over a 9-year period illustrated how new technologies can augment disease surveillance strategies. The number of animal and zoonotic diseases highlights the importance of animals in the study of emerging diseases.


title: Laparoscopic transperitoneal nephrectomy in non-functioning inflammatory kidneys with or without renal stone.

text: OBJECTIVE To compare our laparoscopic simple nephrectomy results in non-functioning inflammatory kidneys with or without renal stones. PATIENTS AND METHODS Patients, who underwent laparoscopic transperitoneal nephrectomy for non-functioning kidney between June, 2010 and October, 2014 were included to study. Overall, data of 32 patients including 15 patients with renal stone (Group 1) and 17 patients without renal stone (Group 2) were retrospectively reviewed. RESULTS Mean age was 44.4 ± 18.5 years (10-71) in group 1 and 35.2 ± 21 years (9-77) in group 2. Mean operation time was 95.0±25.9 minutes (70-175) in group 1 and 86.7 ± 15.1 minutes (70-125) in group 2. Mean estimated blood loss was found to be 64.13 ± 26.67 ml (30-120) in group 1 and 58.94 ± 24.24 ml (30-100) in group 2. Both groups had inflammatory findings in pathological analysis. There was no significant difference between groups regarding estimated blood loss, operation time, pre-operative and post-operative hemoglobin values, percent hemoglobin decrease, complications and hospitalization times (p > 0.05). CONCLUSIONS Non-functioning kidney with or without renal stone could be operated safely with comparable complication rates and success via laparoscopy in experienced hands.


title: Brucella melitensis and Mycobacterium tuberculosis depict overlapping gene expression patterns induced in infected THP-1 macrophages.

text: Pathogens infecting mammalian cells have developed various strategies to suppress and evade their hosts' defensive mechanisms. In this line, the intracellular bacteria that are able to survive and propagate within their host cells must have developed strategies to avert their host's killing attitude. Studying the interface of host-pathogen confrontation can provide valuable information for defining therapeutic approaches. Brucellosis, caused by the Brucella strains, is a zoonotic bacterial disease that affects thousands of humans and animals around the world inflicting discomfort and huge economic losses. Similar to many other intracellular dwelling bacteria, infections caused by Brucella are difficult to treat, and hence any attempt at identifying new and common therapeutic targets would prove beneficial for the purpose of curing infections caused by the intracellular bacteria. In THP-1 macrophage infected with Brucella melitensis we studied the expression levels of four host's genes, i.e. EMP2, ST8SIA4, HCP5 and FRMD5 known to be involved in pathogenesis of Mycobacterium tuberculosis. Our data showed that at this molecular level, except for FRMD5 that was downregulated, the other three genes were upregulated by B. melitensis. Brucella melitensis and M. tuberculosis go through similar intracellular processes and interestingly two of the investigated genes, i.e. EMP2 and ST4SIA8 were upregulated in THP-1 cell infected with B. melitensis similar to that reported for THP-1 cells infected with M. tuberculosis. At the host-pathogen interaction interface, this study depicts overlapping changes for different bacteria with common survival strategies; a fact that implies designing therapeutic approaches based on common targets may be possible.


title: Characterization of a coronavirus isolated from a diarrheic foal.

text: A coronavirus was isolated from feces of a diarrheic foal and serially propagated in human rectal adenocarcinoma (HRT-18) cells. Antigenic and genomic characterizations of the virus (isolate NC99) were based on serological comparison with other avian and mammalian coronaviruses and sequence analysis of the nucleocapsid (N) protein gene. Indirect fluorescent-antibody assay procedures and virus neutralization assays demonstrated a close antigenic relationship with bovine coronavirus (BCV) and porcine hemagglutinating encephalomyelitis virus (mammalian group 2 coronaviruses). Using previously described BCV primers, the N protein gene of isolate NC99 was amplified by a reverse transcriptase PCR (RT-PCR) procedure. The RT-PCR product was cloned into pUC19 and sequenced; the complete N protein of NC99 (446 amino acids) was then compared with published N protein sequences of other avian and mammalian coronaviruses. A high degree of identity (89.0 to 90.1%) was observed between the N protein sequence of NC99 and published sequences of BCV (Mebus and F15 strains) and human coronavirus (strain OC43); only limited identity (<25%) was observed with group 1 and group 3 coronaviruses. Based on these findings, the virus has been tentatively identified as equine coronavirus (ECV). ECV NC99 was determined to have close antigenic and/or genetic relationships with mammalian group 2 coronaviruses, thus identifying it as a member of this coronavirus antigenic group.


title: [Perinatal and neonatal management plan for prevention and control of 2019 novel coronavirus infection (1st Edition)].

text: Since December 2019, the novel coronavirus (2019-nCoV) infection has been prevalent in China. Due to immaturity of immune function and the possibility of mother-fetal vertical transmission, neonates are particularly susceptible to 2019-nCoV. The perinatal-neonatal departments should cooperate closely and take integrated approaches, and the neonatal intensive care unit should prepare the emergency plan for 2019-nCoV infection as far as possible, so as to ensure the optimal management and treatment of potential victims. According to the latest 2019-nCoV national management plan and the actual situation, the Working Group for the Prevention and Control of Neonatal 2019-nCoV Infection in the Perinatal Period of the Editorial Committee of Chinese Journal of Contemporary Pediatrics puts forward recommendations for the prevention and control of 2019-nCoV infection in neonates.


docid: xvdmtedd

title: Infection risk puts the brakes on Canada's biomedical research.


title: Non-structural proteins 2 and 3 interact to modify host cell membranes during the formation of the arterivirus replication complex.

text: The replicase polyproteins of equine arteritis virus (EAV; family Arteriviridae, order Nidovirales) are processed by three viral proteases to yield 12 non-structural proteins (nsps). The nsp2 and nsp3 cleavage products have previously been found to interact, a property that allows nsp2 to act as a co-factor in the processing of the downstream part of the polyprotein by the nsp4 protease. Remarkably, upon infection of Vero cells, but not of BHK-21 or RK-13 cells, EAV nsp2 is now shown to be subject to an additional, internal, cleavage. In Vero cells, approximately 50% of nsp2 (61 kDa) was cleaved into an 18 kDa N-terminal part and a 44 kDa C-terminal part, most likely by a host cell protease that is absent in BHK-21 and RK-13 cells. Although the functional consequences of this additional processing step are unknown, the experiments in Vero cells revealed that the C-terminal part of nsp2 interacts with nsp3. Most EAV nsps localize to virus-induced double-membrane structures in the perinuclear region of the infected cell, where virus RNA synthesis takes place. It is now shown that, in an expression system, the co-expression of nsp2 and nsp3 is both necessary and sufficient to induce the formation of double-membrane structures that strikingly resemble those found in infected cells. Thus, the nsp2 and nsp3 cleavage products play a crucial role in two processes that are common to positive-strand RNA viruses that replicate in mammalian cells: controlled proteolysis of replicase precursors and membrane association of the virus replication complex.


title: Molecular mimicry between Fc receptors and viral antigens.

text: Molecular mimicry has been characterized as the presence of common epitopes, either linear or conformational, shared by host and microbial determinants. Such cross-reactivity may lead to an autoimmune disease. On the other hand molecular mimicry between certain viral proteins and host determinant may protect invading virus to be eliminated by immune system and may promote persistence. In this mini-review I discuss the molecular mimicry of S peplomer protein of mouse hepatitis virus, strain JHM (MHV-JHM) to the host Fc gamma receptor (Fc gamma R). MHV-JHM induces in rodents acute encephalomyelitis and surviving animals develop demyelinating disease with concomitant persistent infection. We have demonstrated that rabbit IgG, but not is F(ab')2 fragments, monoclonal rat and mouse IgG and the rat 2.4G2 anti-Fc gamma R mab immunoprecipitated natural and recombinant S peplomer protein of several strains of MHV. Furthermore, MHV-JHM infected cells formed rosettes with anti-sheep red blood cell (SRBC) - antibody coated SRBC. The 2.4G2 anti-Fc gamma R mab are able to neutralize several strains of MHV, presumably by binding to S peplomer protein. Therefore, the Fc binding site of S is present on the surface of MHV-infected cells. This molecular mimicry between S peplomer protein of MHV-JHM and Fc gamma R has been extended to other members of Coronaviridae, namely bovine coronavirus and transmissible gastroenteritis virus but not to infectious bronchitis virus. The molecular mimicry of viral antigens to Fc receptors has been described also for members of Herpesviridae, namely Herpes simplex, cytomegalovirus and Varicella zoster.(ABSTRACT TRUNCATED AT 250 WORDS)


Laughter is the best medicine.


docid: mtanqncf

title: Postoperative non-steroidal anti-inflammatory drugs and colorectal anastomotic leakage. NSAIDs and anastomotic leakage.

text: Anastomotic leakage (AL) is the most important and one of the most serious complications after colorectal resections with primary anastomosis. Any factors that contribute to increase the risk of AL should be identified and--if possible--eliminated. Non-steroidal anti-inflammatory drugs (NSAIDs) are often used for treating pain after surgical procedures, among these also colorectal resections. The objective of this Ph.d. thesis was to investigate whether the use of NSAIDs in the postoperative period increases the risk of AL, and investigate the effect on pathophysiological mechanisms. In order to achieve this, the following studies were performed. Study I was a retrospective, case-control study in 75 patients undergoing laparoscopic colorectal resection for colorectal cancer. 33 of these patients received the NSAID diclofenac in the postoperative period; the remaining 42 did not receive any NSAID. There were significantly more ALs among the patients receiving diclofenac (7/33 vs. 1/42, p=0.018). In uni- and multivariate logistic regression analyses, diclofenac was the only factor associated with increased AL rate. This study functioned as a hypothesis generating study and laid the ground for the subsequent studies. Study II was an experimental, randomized, case-control study in 32 Wistar rats. The rats had a colonic anastomosis performed and were randomized to diclofenac or placebo treatment. After three days, the rats were sacrificed and the anastomoses were harvested. First, the anastomotic strengths were tested by longitudinal; subsequently, the levels of the enzyme cyclooxygenase-2 (COX-2) in the anastomotic tissues were measured. There was no difference among the groups with regard to anastomotic strength, but the animals treated with diclofenac had significantly lower COX-2 levels (median (range) 1.30 (0.42-3.31) ng/mg vs. 2.44 (0.88 - 18.94) ng/mg, p<0.001). This study showed that the used dose of diclofenac was sufficient and relevant, but did not show a direct damaging effect on the anastomoses due to NSAID treatment. Study III was also an experimental, randomized, case-control study. This time round, 60 Wistar rats were included. Again, colonic anastomoses were performed and the rats were randomized to diclofenac or placebo. Also, expanded polytetrafluoruethylene (ePTFE) tubes were placed under the skin of the rats. In this material, substituents of connective tissue accumulate and the amount of accumulation can be measured. After 7 days, the rats were sacrificed and, again, anastomotic strengths were measured along with collagen content in the ePTFE tubes. Anastomotic strength was similar in the two groups while collagen accumulation was significantly decreased among the rats treated with diclofenac (median (i.q.r.) 0.29 (0.13-0.47) vs. 0.47 (0.28-0.62) mcg/mg, p = 0.03). This study for the first time showed that NSAID inhibit subcutaneous collagen formation and that this formation is reversely correlated to anastomotic strength. This information can be used in further studies in this subject. Study IV was the final experimental case-control study in 40 Wistar rats. This time, in order to more easily extrapolate experimental results to daily clinical life, the colonic anastomoses were sutured with the same type of suture material as used in the clinical setting. Thus, half the anastomoses was performed with resorbable suture; the other half with non-resorbable suture. None of the rats received NSAID. The breaking strength was compared and found similar in the two groups. This study showed that experimental studies can be optimized in order to make comparisons and extrapolations to the clinical setting easier. Study V was a database study based on data from the Danish Colorectal Cancer Group's (DCCG) prospective database and electronically registered medical records. From the database information on demographic, surgical and postoperative variables (including AL) were provided. Information on NSAID consumption was retrieved by individual searches in the patients' medical records. Based on these data, uni- and multivariate logistic regression analyses were performed. These analyses identified NSAID treatment in the postoperative period as an individual risk factor for AL. Other risk factors identified were consistent with the available literature. The detrimental effect of the NSAIDs are possibly due to an effect on collagen metabolism leading to weakened tissue around the anastomosis and/or on the risk of thrombosis formation leading to more thromboses in the vessels supplying the anastomosis, thereby limiting anastomotic blood flow. In conclusion, the studies included in this thesis have elucidated some of the physiological and pathophysiological mechanisms involved in anastomotic healing and leakage, and furthermore have shown that the use of NSAIDs in the postoperative period increase the risk of AL in patients undergoing colorectal surgery with primary anastomosis. Based on the findings in these studies, and based on existing knowledge, it is recommended that NSAIDs be abandoned after colorectal resection with primary anastomosis. It should be investigated whether the NSAIDs are also harmful to other types of anastomoses and after other surgical procedures where early tissue healing is crucial.


The impact of the intracerebral antibody response on the clinical course of a virus-induced demyelination in a rat model system.


title: Respiratory consequences of rhinovirus infection.

text: Rhinoviruses, a genus of the family Picornaviridae, are the cause of more than 50% of respiratory tract infections. Complications of rhinovirus infections, which include otitis media, sinusitis, exacerbations of asthma, and other pulmonary diseases, can be significant in certain populations. Reverse transcriptase-polymerase chain reaction has allowed the identification of rhinoviruses and led to a greater appreciation of the role of this pathogen in upper and lower respiratory tract disease. Furthermore, antiviral agents with broad activity against rhinoviruses have recently been developed, have undergone clinical trials, but have not been approved for clinical use. By altering the clinical course of picornavirus infections, it may be possible to minimize their potential adverse consequences.


docid: obckxglw

title: Emerging zoonoses and pathogens of public health significance--an overview.

text: Emerging zoonotic diseases have assumed increasing importance in public and animal health, as the last few years have seen a steady stream of new diseases, each emerging from an unsuspected quarter and causing severe problems for animals and humans. The reasons for disease emergence are multiple, but there are two main factors--expansion of the human population and globalisation of trade. Current issues such as the increasing movement of a variety of animal species, ecological disruption, uncultivatable organisms, and terrorism, all imply that emerging zoonotic diseases will in all probability, not only continue to occur, but will increase in the rate of their emergence. The recurring nature of the crises dictates that closer integration of veterinary and medical communities is warranted, along with improved education of the general public and policy makers.


title: In vitro characterization of TMPRSS2 inhibition in IPEC-J2 cells.

text: The transmembrane serine protease, TMPRSS2 is an important target in the treatment of seasonal influenza infections and contributes to prostate carcinogenesis and metastasis. In this study, the effect of the synthetic TMPRSS2 inhibitor I-432 on jejunal IPEC-J2 cell monolayers cultured on membrane inserts was characterized. Using a fluorogenic substrate, it was found that the apical addition of I-432 could suppress trypsin-like activity in the supernatants of IPEC-J2 cells. The inhibition of TMPRSS2 did not affect physiologically produced hydrogen peroxide levels in the apical and in basolateral compartments. Loss of expression of the TMPRSS2 serine protease domain (28 kDa) was also observed when cells were pre-exposed to I-432. Partial decrease in immunofluorescent signal intensities derived from the altered distribution pattern of TMPRSS2 was detected after a 48 h long incubation of IPEC-J2 cells with the inhibitor indicating the efficacy of TMPRSS2 inhibition via I-432 administration in vitro.


title: Suppression of viral RNA recombination by a host exoribonuclease.

text: RNA viruses of humans, animals, and plants evolve rapidly due to mutations and RNA recombination. A previous genome-wide screen in Saccharomyces cerevisiae, a model host, identified five host genes, including XRN1, encoding a 5'-3' exoribonuclease, whose absence led to an approximately 10- to 50-fold enhancement of RNA recombination in Tomato bushy stunt virus (E. Serviene, N. Shapka, C. P. Cheng, T. Panavas, B. Phuangrat, J. Baker, and P. D. Nagy, Proc. Natl. Acad. Sci. USA 102:10545-10550, 2005). In this study, we found abundant 5'-truncated viral RNAs in xrn1delta mutant strains but not in the parental yeast strains, suggesting that these RNAs might serve as recombination substrates promoting RNA recombination in xrn1delta mutant yeast. This model is supported by data showing that an enhanced level of viral recombinant accumulation occurred when two different 5'-truncated viral RNAs were expressed in the parental and xrn1delta mutant yeast strains or electroporated into plant protoplasts. Moreover, we demonstrate that purified Xrn1p can degrade the 5'-truncated viral RNAs in vitro. Based on these findings, we propose that Xrn1p can suppress viral RNA recombination by rapidly removing the 5'-truncated RNAs, the substrates of recombination, and thus reducing the chance for recombination to occur in the parental yeast strain. In addition, we show that the 5'-truncated viral RNAs are generated by host endoribonucleases. Accordingly, overexpression of the Ngl2p endoribonuclease led to an increased accumulation of cleaved viral RNAs in vivo and in vitro. Altogether, this paper establishes that host ribonucleases and host-mediated viral RNA turnover play major roles in RNA virus recombination and evolution.


MR diffusion-weighted imaging in a case of West Nile virus encephalitis.


Connection and communication vital to handling a pandemic in Ontario.


title: New TMA Leaders Chosen Amid Historic Pandemic.

text: Despite the suspension of the Texas Medical Association House of Delegates due to the COVID-19 pandemic, special provisions in TMA's bylaws allowed for limited elections to take place on May 2, including that of Edinburg internist E. Linda Villarreal, MD, as president-elect. And Diana L. Fite, MD, took office as planned as TMA's 155th president.


title: [Detection and clinical analysis of acute lower respiratory tract infection with human coronaviruses in children in Beijing area 2007-2015].

text: OBJECTIVE To investigate human coronaviruses (HCoVs) infection in children with acute lower respiratory tract infection(ALRTI)and to explore the clinical features of ALRTI caused by HCoVs in children. METHOD Totally 4 371 children with clinical diagnosis of ALRTI during the period from March 2007 to February 2015 seen in Beijing Children's Hospital were recruited into this study. Patients were divided into 4 groups by age, including 1 890 cases in < 1 year group, 788 cases in 1-3 years group, 553 cases in 3-6 years group, 1140 cases in ≥6 years group. One nasopharyngeal aspirate specimen was collected from each patient. RT-PCR methods were applied to detect 9 common respiratory viruses including HCoVs (including HCoV-OC43, HCoV-229E, HCoV-NL63 and HCoV-HKU1), respiratory syncytial virus (RSV) and so on. Clinical features of ALRTI with single HCoVs infection were analyzed and compared with hospitalized ALRTI cases with single RSV infection in the same period. RESULT (1) Totally 2 895 cases were positive for at least one virus in this study in 4 371 ALRTI patients (positive rate 66.23%), in which 147 cases were positive for HCoVs infection (positive rate 3.36%). (2) Positive rates of HCoVs in each year from 2007 to 2014 were 6.11%, 3.79%, 4.69%, 4.31%, 2.38% 2.10%, 0.77% and 2.65%, respectively. The mean positive rates of HCoVs for each month from January to December were 2.53%, 2.12%, 3.63%, 6.68%, 1.53%, 3.77%, 3.92%, 3.00%, 2.15%, 5.26%, 3.01% and 2.80%. (3) Detection results of each subtypes of HCoVs in total 4 371 pediatric ALRTI patients were: 48 cases positive for HCoV-OC43(1.10%), 32 cases positive for HCoV-229E(0.73%), 25 cases positive for HCoV-NL63 (0.57%), 27 cases positive for HCoV-HKU1 (0.62%). (4) Positive rates of HCoVs infection in <1 year group, 1-3 years group, 3-6 years group and ≥ 6 years group were 4.13%, 5.08%, 2.71% and 1.23%, respectively. There were significant differences in positive rates of HCoV among groups (χ² = 27.218, P<0.01). (5) There were 16 hospitalized cases with single infection of HCoVs in this study, of which 12 cases were diagnosed as bronchopneumonia, 3 cases developed acute laryngeal obstruction, 2 cases had acute bronchial asthma attack. Common clinical manifestations included cough (14 cases), gasping (13 cases), dyspnea (9 cases), fever (6 cases), hoarseness (4 cases), laryngeal stridor (4 cases) and abnormality on chest X-ray (including fuzzy lung texture, patchy shadow and consolidation) (12 cases). (6) There were no significant differences in the incidence of clinical manifestations (including cough, gasping, dyspnea, fever and abnormality on chest X-ray), complications (including respiratory failure, myocardial damage, and acute bronchial asthma attack) and mechanical ventilation between hospitalized ALRTI patients with single HCoV infection and 193 patients with single RSV infection in the same period. CONCLUSION HCoVs are pathogens of ALRTI in children, The overall positive rate of HCoVs was 3.36% in this study. The clinical manifestations and severity of ALRTI caused by single HCoVs was comparable to that of ALRTI with single RSV infection in children.


title: Detection of severe acute respiratory syndrome coronavirus in blood of infected patients.

text: Severe acute respiratory syndrome (SARS) has caused major outbreaks worldwide, resulting in an urgent need to obtain sensitive and accurate diagnosis of this disease. PCR-based detection methods were developed for use on a variety of samples, including blood. Eighteen subjects were investigated, and results indicated that blood samples contain sufficient virus for detection by using quantitative real-time PCR.


title: Structural requirements for O-glycosylation of the mouse hepatitis virus membrane protein.

text: The mouse hepatitis virus (MHV) membrane (M) protein contains only O-linked oligosaccharides. We have used this protein as a model to study the structural requirements for O-glycosylation. We show that MHV M is modified by the addition of a single oligosaccharide side chain at the cluster of 4 hydroxylamino acids present at its extreme amino terminus and identified Thr at position 5 as the functional acceptor site. The hydroxylamino acid cluster, which is quite conserved among O-glycosylated coronavirus M proteins, is not in itself sufficient for O-glycosylation. Downstream amino acids are required to introduce a functional O-glycosylation site into a foreign protein. In a mutagenic analysis O-glycosylation was found to be sensitive to some particular changes but no unique sequence motif for O-glycosylation could be identified. Expression of mutant M proteins in cells revealed that substitution of any 1 residue was tolerated, conceivably due to the occurrence of multiple UDP-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltransferases (GalNAc transferases). Indeed, MHV M served as a substrate for GalNac-T1, -T2, and -T3, as was demonstrated using an in situ glycosylation assay based on the co-expression of endoplasmic reticulum-retained forms of the GalNAc transferases with endoplasmic reticulum-resident MHV M mutants. The GalNAc transferases were found to have largely overlapping, but distinct substrate specificities. The requirement for a threonine as acceptor rather than a serine residue and the requirement for a proline residue three positions downstream of the acceptor site were found to be distinctive features.


title: Axium MicroFX coil for the completing endovascular aneurysm surgery study (ACCESS). A prospective evaluation of the safety and durability of Axium MicroFX PGLA coils.

text: Recanalization of previously coiled aneurysms remains a major drawback of endovascular aneurysm therapy. We performed a prospective single arm trial to provide early initial data regarding the safety and angiographic durability of a new coil technology, the Axium MicroFX Polyglycolic/polylactic acid (PGLA) coil, which was designed to lower recanalization rates. Fifteen patients (16 aneurysms) were prospectively enrolled. Demographic and peri-procedural data were collected. Angiographic images of the initial coil embolization and three to six month follow-up angiographic images underwent blinded evaluation. Seven (47%) SAH and eight (53%) elective patients were enrolled. Blinded evaluation of the initial embolization demonstrated that 5/16 (31%) aneurysms achieved Raymond grade 1, 5/16 (31%) grade 2 and 6/16 (38%) grade 3. Three to six month angiography was obtained in 12/15 patients (80%); two patients expired (1 SAH, 1 elective) and one was lost to follow-up (SAH). All patients who underwent follow-up angiography had a mRS ≤1. Blinded evaluation of embolization demonstrated 7/13 aneurysms (54%) improved in Raymond grading, five (38%) were stable and one aneurysm (8%) worsened. One patient developed an asymptomatic peri-aneurysmal parent vessel stenosis. Axium MicroFX coils appear to be safe, though the small number of patients in this series obviates comparative analysis with other series. Further studies are needed with more patients to compare the angiographic durability of Axium MicroFX coils to other coils.


title: [Severe acute respiratory syndrome (SARS)].

text: The severe acute respiratory syndrome (SARS) is a highly infectious respiratory disease, to the best of our knowledge caused by a hitherto unknown corona virus. The virus has spread from South East Asia to many countries of the world. Three case reports of patients from the Prince of Wales Hospital of The Chinese University of Hong Kong demonstrate typical clinical courses. Fever, cough, in most cases non-productive, myalgia, chills, and rigor are the leading symptoms. Leucopenia and thrombocytopenia are the most prominent laboratory parameters, increased values for lactatedehydrogenase (LDH) reflect a more severe clinical course. Advanced age and coexisting conditions seem to influence the prognosis unfavourably. The chest roentgenogram may be normal initially but at a later stage progressive consolidations in the majority of peripheral parts of the lung are observed, which cannot be differentiated from pneumonias of other origin. Even young patients can enter a stage of respiratory compromise rather fast. A therapy against the cause of the disease is not known. Empirical therapy with ribavirin in combination with high dose corticosteroids have proved successful. The disease may progress into respiratory failure comparable with an acute respiratory distress syndrome (ARDS). Mortality is around five to ten per cent. Stringent hygiene and quarantine measures are mandatory to prevent the further spread of this threatening disease.


title: [Management of COVID-19: the Zhejiang experience].

text: The current epidemic situation of coronavirus disease 2019 (COVID-19) still remained severe. As the National Clinical Research Center for Infectious Diseases, the First Affiliated Hospital of Zhejiang University School of Medicine is the primary medical care center for COVID-19 in Zhejiang province. Based on the present expert consensus carried out by National Health Commission and National Administration of Traditional Chinese Medicine, our team summarized and established an effective treatment strategy centered on "Four-Anti and Two-Balance" for clinical practice. The "Four-Anti and Two-Balance" strategy included antivirus, anti-shock, anti-hyoxemia, anti-secondary infection, and maintaining of water, electrolyte and acid base balance and microecological balance. Meanwhile, integrated multidisciplinary personalized treatment was recommended to improve therapeutic effect. The importance of early viralogical detection, dynamic monitoring of inflammatory indexes and chest radiograph was emphasized in clinical decision-making. Sputum was observed with the highest positive rate of RT-PCR results. Viral nucleic acids could be detected in 10%patients' blood samples at acute period and 50%of patients had positive RT-PCR results in their feces. We also isolated alive viral strains from feces, indicating potential infectiousness of feces.Dynamic cytokine detection was necessary to timely identifying cytokine storms and application of artificial liver blood purification system. The "Four-Anti and Two-Balance" strategy effectively increased cure rate and reduced mortality. Early antiviral treatment could alleviate disease severity and prevent illness progression, and we found lopinavir/ritonavir combined with abidol showed antiviral effects in COVID-19. Shock and hypoxemia were usually caused by cytokine storms. The artificial liver blood purification system could rapidly remove inflammatory mediators and block cytokine storm.Moreover, it also favored the balance of fluid, electrolyte and acid-base and thus improved treatment efficacy in critical illness. For cases of severe illness, early and also short period of moderate glucocorticoid was supported. Patients with oxygenation index below 200 mmHg should be transferred to intensive medical center. Conservative oxygen therapy was preferred and noninvasive ventilation was not recommended. Patients with mechanical ventilation should be strictly supervised with cluster ventilator-associated pneumonia prevention strategies. Antimicrobial prophylaxis was not recommended except for patients with long course of disease, repeated fever and elevated procalcitonin (PCT), meanwhile secondary fungal infection should be concerned.Some patients with COVID-19 showed intestinal microbial dysbiosis with decreased probiotics such as Lactobacillus and Bifidobacterium, so nutritional and gastrointestinal function should be assessed for all patients.Nutritional support and application of prebiotics or probiotics were suggested to regulate the balance of intestinal microbiota and reduce the risk of secondary infection due to bacterial translocation. Anxiety and fear were common in patients with COVID-19. Therefore,we established dynamic assessment and warning for psychological crisis. We also integrated Chinese medicine in treatment to promote disease rehabilitation through classification methods of traditional Chinese medicine. We optimized nursing process for severe patients to promote their rehabilitation. It remained unclear about viral clearance pattern after the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Therefore, two weeks' quarantine for discharged patients was required and a regular following up was also needed.The Zhejiang experience and suggestions have been implemented in our center and achieved good results. However, since COVID-19 was a newly emerging disease, more work was warranted to improve strategies of prevention, diagnosis and treatment for COVID-19.


title: Workload and prevalence of open wounds in the community: French Vulnus initiative.

text: OBJECTIVE To calculate the prevalence of open cutaneous wounds presented on a routine working day in community settings in metropolitan France, and to estimate the workload associated with the care of these wounds by nurses, GPs and specialists (dermatologists, diabetologists and phlebologists). METHOD A transversal epidemiological survey was conducted on a randomly selected sample of the above practitioners between June and July 2008. The percentage of patients presenting on a routine working day with open a cutaneous lesion of any origin, location, size and duration was assessed. All local and systemic care performed on the patient during that day because of the wound was also recorded. RESULTS In total, 475 GPs, 453 specialists and 238 nurses participated (n=1166) and saw a total of 29 663 patients, of whom 3037 presented with one or more cutaneous wound. The overall non-weighted prevalence of patients with a wound was 10.2% (95%CI: 9.9%;10.6%). This prevalence was similar for GPs (6.0%) and for specialists (6.9%), but was higher for nurses (22.0%). Forty-three per cent of all wounds had a duration of over 6 weeks. These chronic wounds were predominantly leg ulcers, diabetic foot ulcers or pressure ulcers, but also included wounds of all aetiologies. For 33% of all patients with wounds, the impact on their health status was serious to severe. The overwhelming majority of wounds (95%) required local care, including in 65% of cases cleansing and debridement. CONCLUSION Despite its limitations, this initiative, the first of its type in France, strongly suggests that wound care constitutes an important part of routine care given by health professionals in the community, and for a substantial number of these patients, wounds represent a serious morbidity. DECLARATION OF INTEREST The non-profit organisation 'Association Vivre avec une Plaie' financially supported this study. This association received unrestricted grants from the French Wound Healing Society (SFFPC) and a consortium of private companies (main sponsors: ConvaTec, Genevrier, Hartmann, KCI, Mölnycke, Smith & Nephew, Urgo; minor sponsors: Coloplast, Covidien, HNE) to fund the costs incurred by the methodological process and statistical analysis but had no input into the findings. The National Health Insurance Organisation provided non-financial support to this initiative. J.C. Kerihuel received support for the submitted work from 'Association Vivre avec une Plaie'. S. Meaume, I. Fromantin and L. Téot have no financial relationship with 'Association Vivre avec une Plaie', for either this or any work submitted in the previous 3 years. The authors have no non-financial interests that may be relevant to the submitted work, and their spouses, partners, or children have no financial relationships that may be relevant to the submitted work


title: Coronavirus-like particles as aetiological agents of acute non-bacterial gastroenteritis in humans.

text: The morphology of coronavirus-like particles found in faecal extracts of neonates and infants with acute gastroenteritis show similarities with the structure of human coronaviruses. These coronavirus-like particles can be detected in faeces of sick neonates and infants in higher frequency than in healthy children of the same age, their association with this clinical syndrome is discussed. The significance of enteric coronavirus-like particles in acute gastroenteritis of adults is questionable since the particles can be found in faeces of diseased persons and of healthy adults with equal frequency.


title: Bovine Cryptosporidiosis: Clinical and Pathological Findings in Forty-two Scouring Neonatal Calves.

text: Cryptosporidia organisms were identified in 42 of 161 (26%) neonatal, diarrheic calves, over a 32 month period commencing July 1979. Forty of the 161 calves were submitted alive and cryptosporidiosis was diagnosed in 63% (25 of 40) of them. The cryptosporidia infected calves were usually one to two weeks old and came from 26 herds where the typical history was profuse, watery diarrhea in nearly all neonatal calves. The diarrhea usually started around one week of age, was unresponsive to all conventional antidiarrhea therapies, lasted for two or more weeks and was usually fatal. Twenty-nine (69%) of the cryptosporidia infected calves were submitted between December and February. These calves were often hutch reared.Histopatholoical examination revealed large numbers of the coccidial parasite Cryptosporidium sp embedded in the microvilli of jejunal and ileal absorptive enterocytes of all affected calves. The organisms were identified as trophozoites and schizonts (asexual stages) and macrogametes (female sexual stages) with the electron microscope. Microgametes (male sexual stages) were not identified. Occasionally a merozoite (asexual stage) was also seen apparently burrowing into or about to be enveloped by a host microvillus. Observation of the organisms was much easier when diarrheic calves were submitted alive. Enterotoxigenic Escherichia coli were often cultured from intestines of dead calves and occasionally from calves submitted alive. Coronavirus particles were seen in one calf. In the last year of this study, oocysts were identified in fecal smears stained with May-Grünwald-Giemsa stain and fecal samples using a dichromate solution flotation technique.


title: [Prevalence of burnout syndrome in nursing staff in a large hospital in south of Brazil].

text: Burnout syndrome is a chronic adaptive disorder that affects workers and is characterized by three forms of psychological distress: emotional exhaustion, depersonalization, and a sense of low personal fulfillment. A cross-sectional quantitative design was used to apply questionnaires including socio-cultural data and the Maslach Burnout Inventory in 151 nurses and nurse technicians, representing 54.1% of this professional category in a large general hospital in Tubarão, Santa Catarina State, Brazil. Mean scores were: emotional exhaustion (17), depersonalization (7.79), and low personal fulfillment (36.6). According to the criterion proposed by Ramirez et al., no cases of burnout were diagnosed. According to the criteria by Grunfeld et al., 35.7% of the interviewees displayed burnout. The standard profile of workers with burnout identified in the study was: nurse technicians, female gender, age 26-35 years, married, with no children, and with five or more years in the profession. Hospital wards or areas with the highest proportion of nursing staff with burnout were the grouped hospital sectors (42.6%), ICU (25.9%), and neonatal ICU (18.5%). There was no statistically significant association between ward or hospital area and burnout.


The SARS epidemic in Hong Kong.


title: Serum respiratory virus antibodies: predictor of reduced one-second forced expiratory volume (FEV1) in Norwegian adults.

text: BACKGROUND The purpose of this cross-sectional study was to investigate whether the presence of serum complement antibodies was associated with reduced one-second forced expiratory volume (FEV1) in adults. METHODS From a stratified random sample of 18-73 year old adults, we performed measurements of serum complement fixing virus antibodies against influenza type A and B, parainfluenza type 1, 2, and 3, respiratory syncytial virus and adenovirus on 82% (n = 1239). RESULTS In the crude data, subjects having five of the seven virus antibodies had significantly lower lung function, given as sex-, age- and height-standardized residuals of FEV1 (SFEV1), compared with those without. After adjusting in addition for smoking habits, lifetime smoking consumption and season, the lung function levels were significantly lower in subjects with influenza type B and respiratory syncytial virus antibodies compared to those without (P < 0.01). Increasing influenza and respiratory syncytial virus antibody titres and increasing numbers of virus antibodies, respectively, were related to progressively lower lung function. Subjects with respiratory symptoms but without obstructive lung disease had lower antibody levels than subjects with obstructive lung disease, but higher levels than asymptomatic subjects. In a final multiple linear regression analysis adjusting in addition for respiratory symptom and disease status as well as for the other respiratory virus antibodies, the presence of respiratory syncytial virus antibodies was a significant predictor for reduced SFEV1 (regression coefficient: -0.226; SE = 0.112; P = 0.04). The magnitude of the effect on lung function remained after excluding subjects reporting symptoms of respiratory infection within 3 weeks prior to the examination (regression coefficient: -0.252; SE = -0.218; P = 0.25). CONCLUSIONS This cross-sectional community study indicates that respiratory syncytial virus infection or re-infection is an independent predictor for reduced lung function in adults of a wide age range.


docid: qzjbxofn

title: The role of PARP inhibitors in germline BRCA-associated pancreatic ductal adenocarcinoma.

text: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy that remains a challenge to treat. In pursuit of personalized medicine, researchers continue active exploration of the genetic and molecular framework of PDAC to apply novel therapeutics and enhance outcomes. In patients who have PDAC, germline mutations-such as those in the BRCA1/2 and PALB2 genes-are predominantly associated with the DNA damage response and repair pathway. On the basis of studies completed in patients with BRCA-mutated advanced breast and ovarian cancer, the poly(ADP-ribose) polymerase (PARP) inhibitors have been evaluated for safety, tolerability, and efficacy in patients with advanced PDAC who are carrying germline BRCA gene mutations. Results have demonstrated meaningful activity and identified BRCA as a predictive and targetable biomarker in PDAC, and have also identified the role of olaparib as a maintenance therapy in PDAC. On the basis of the principle of synthetic lethality, and to avert resistance to PARP inhibitors, clinical trials of combination therapy with PARP inhibitors and platinum-based chemotherapy have been conducted with an early signal. As we continue to explore the role of PARP inhibitors in the management of PDAC, recent clinical trials are studying the effectiveness of PARP inhibitors in combination with immunotherapy, targeted inhibitors, and angiogenesis inhibitors. The next steps are to understand the role of PARP inhibitors beyond germline BRCA in other homologous recombination repair gene mutations and in other subgroups of patients with PDAC.


title: Neurotropism of human coronavirus 229E.

text: The 299E prototype strain of human coronavirus (HCV-229E) has so far been mainly associated with infections of the respiratory tract. In the present study, we show evidence for infection of the central nervous system (CNS) by HCV-229E, both in vitro and in vivo. Various human cell lines of CNS origin were tested for their susceptibility to infection by HCV-229E. Production of viral antigens was monitored by indirect immunofluorescence with monoclonal antibodies and infectious progeny virions by plaque assay on the L132 human embryonic lung cell line. The SK-N-SH neuroblastoma and H4 neuroglioma cell lines were highly susceptible to infection. The U-87 MG and U-373 MG astrocytoma cell lines were also infectable by HCV-229E. We could also demonstrate infection of the MO3.13 cell line, which was established by fusion of human oligodendrocytes with a thioguanine-resistant mutant of the TE671 (RD) human rhabdomyosarcoma cell line. An apparently more extensive infection of the MO3.13 cells, when compared to the parental cells, supports the notion that human oligodendrocytes are differentially susceptible to infection by this virus. We also tested for HCV-229E gene expression in pathological brain specimens. For that purpose, we developed a reverse transcription-polymerase chain reaction (RT-PCR) assay to amplify a portion of the mRNA encoding the viral nucleocapsid protein. Using stringent laboratory conditions, viral RNA was detectable in brain tissue of 4 of 11 multiple sclerosis patients and none of 6 neurological and 5 normal controls.(ABSTRACT TRUNCATED AT 250 WORDS)


The COVID-19 Intubation and Ventilation Pathway (CiVP); a Commentary.


title: Host deadenylation-dependent mRNA decapping factors are required for a key step in brome mosaic virus RNA replication.

text: The genomes of positive-strand RNA [+RNA] viruses perform two mutually exclusive functions: they act as mRNAs for the translation of viral proteins and as templates for viral replication. A universal key step in the replication of +RNA viruses is the coordinated transition of the RNA genome from the cellular translation machinery to the viral replication complex. While host factors are involved in this step, their nature is largely unknown. By using the ability of the higher eukaryotic +RNA virus brome mosaic virus (BMV) to replicate in yeast, we previously showed that the host Lsm1p protein is required for efficient recruitment of BMV RNA from translation to replication. Here we show that in addition to Lsm1p, all tested components of the Lsm1p-7p/Pat1p/Dhh1p decapping activator complex, which functions in deadenylation-dependent decapping of cellular mRNAs, are required for BMV RNA recruitment for RNA replication. In contrast, other proteins of the decapping machinery, such as Edc1p and Edc2p from the deadenylation-dependent decapping pathway and Upf1p, Upf2p, and Upf3p from the deadenylation-independent decapping pathway, had no significant effects. The dependence of BMV RNA recruitment on the Lsm1p-7p/Pat1p/Dhh1p complex was linked exclusively to the 3' noncoding region of the BMV RNA. Collectively, our results suggest that the Lsm1p-7p/Pat1p/Dhh1p complex that transfers cellular mRNAs from translation to degradation might act as a key regulator in the switch from BMV RNA translation to replication.


title: Astrovirus ribosomal frameshifting in an infection-transfection transient expression system.

text: Different regions of the human astrovirus frameshift signal were cloned into the rhesus rotavirus VP4 gene and evaluated in an infection-transfection transient expression cell culture system. BHK-21 cells, infected with a vaccinia virus that expresses T7 RNA polymerase (vTF7-3), were transfected with the various astrovirus-VP4 constructs. All constructs were driven by a T7 promoter and contained an internal ribosome entry site. Frameshifted and nonframeshifted protein products were immunoprecipitated with VP4 amino- and carboxy-terminal-specific monoclonal antibodies, and their ratios were determined by PhosphorImager analysis. The efficiency of frameshifting was 25 to 28%, significantly greater than the 5 to 7% efficiency reported previously in a cell-free translation system. Coupling of transcription and translation in a cell-free system yielded frameshifting efficiencies threefold greater than that of the uncoupled in vitro system. The presence of the shifty heptamer was an absolute requirement for frameshifting in both cell-free and intact-cell systems, while deletion of the potential downstream pseudoknot region did not affect the efficiency of frameshifting.


title: A new paradigm for quarantine and public health activities at land borders: opportunities and challenges.

text: The Institute of Medicine (IOM) report Quarantine Stations at Ports of Entry: Protecting the Public's Health focused almost exclusively on U.S. airports and seaports, which served 106 million entries in 2005. IOM concluded that the primary function of these quarantine stations (QSs) should shift from providing inspection to providing strategic national public health leadership. The large expanse of our national borders, large number of crossings, sparse federal resources, and decreased regulation regarding conveyances crossing these borders make land borders more permeable to a variety of threats. To address the health challenges related to land borders, the QSs serving such borders must assume unique roles and partnerships to achieve the strategic leadership and public health research roles envisioned by the IOM. In this article, we examine how the IOM recommendations apply to the QSs that serve the land borders through which more than 319 million travelers, immigrants, and refugees entered the U.S. in 2005.


title: [Expression of the recombinant SARS coronavirus nucleocapsid protein in Pichia pastoris and identification of its bioactivity].

text: The full length cDNA of SARS coronavirus nucleocapsid (N) protein was amplified by PCR and cloned into yeast expression vector pPIC3.5K to generate expression vector pPIC3.5K-SCoVN. The plasmid was linearized and then transformed into P. pastoris (His- Mut+) by electroporation method. His+ Mut+ recombinant strains were screened on G418-RDB and MM/MD plates, and further confirmed by PCR. The influence of various inducing media, dissolved oxygen(DO) and the different final concentration of methanol was subsequently investigated. The results showed that the FBS medium was optimal for recombinant N protein expression and growth of the recombinant strain. The optimal final concentration of methanol is 1% (V/V), and the DO has a significant effect on recombinant N protein expression and growth of recombinant strain. The recombinant N protein expressed was about 6% of the total cell proteins, 410 mg/L of recombinant N protein and 45 OD600 were achieved in shake flask. Western-blot showed that the recombinant N protein had high specificity against mouse-anti-N protein-mAb and SARS positive sera, but had no cross-reaction with normal human sera. The result of scale-up culture in fermemtator demonstrated that 2.5g/L of recombinant N protein and the maximum cell 345 OD600 of were achieved, which was 6.1 times and 7.7 times higher than that in shake flask. So this study provide a basis for further researches on the early diagnosis of SARS and the virus reproduction and pathology reaction of SARS coronavirus.


title: Consistency and proportionality in policy decision-making in blood safety: the case for an all-apheresis platelet supply in Germany.

text: Recently, German investigators presented the first mathematical model finding a significant increase in the risk of HIV, HCV, and HBV transmission when pools of 4 whole-blood-derived buffy-coat platelets, rather than 1 single-donor (apheresis) component, are used to provide one platelet dose. Based, in both cases, on mathematical models employing the incidence/window-period method, the relative risk of transmission from pooled versus apheresis platelets (2.2 or 2.75 for HIV, 2.7 or 3.375 for HCV, and 3.2 or 4.0 for HBV, with pools of 4 or 5 concentrates, respectively) is similar to the difference in risk before (versus after) introduction of HIV-1 and HCV RNA screening. The absolute increase in the risk from pools (1 to 2 HIV-, HCV-, or HBV-infectious platelet doses annually) is much smaller than the yield from HIV-1 and HCV RNA screening projected in the 1990s, but it becomes similar to that yield (with up to 88 infectious platelet doses intercepted) when we consider the next transfusion-transmitted pathogen to emerge in the future. Although pathogen reduction (PR) of platelets would eliminate the difference in risk between pooled and apheresis platelets vis-a-vis viral transmission, PR is not ready for implementation because the safety of PR needs to be investigated further. German transfusion guidelines should be revised to indicate the difference in risk associated with pooled versus apheresis platelets, and transition toward an all-apheresis platelet supply should commence. These actions are consistent with and proportionate to the action taken in the 1990s when screening for HIV-1 and HCV RNA was implemented.


title: Endovascular treatment of unruptured aneurysms of cavernous and ophthalmic segment of internal carotid artery with flow diverter device Pipeline.

text: BACKGROUND Intra-arterial treatment of aneurysms by redirecting blood flow is a newer method. The redirection is based on a significantly more densely braided wire stent. The stent wall keeps the blood in the lumen of the stent and slows down the turbulent flow in the aneurysms. Stagnation of blood in the aneurysm sac leads to the formation of thrombus and subsequent exclusion of the aneurysm from the circulation. The aim of the study was to evaluate flow diverter device Pipeline for broad neck and giant aneurysm treatment. METHODS Fifteen patients with discovered aneurysm of the internal carotid artery were treated between November 2010 and February 2014. The majority of aneurysms of the internal carotid artery were located intradural at the ophthalmic part of the artery. The patients were treated using a flow diverter device Pipeline, which was placed over the aneurysm neck. Treatment success was assessed clinically and angiographically using O'Kelly Marotta scale. RESULTS Control angiography immediately after the release of the stent showed stagnation of the blood flow in the aneurysm sac. In none of the patients procedural and periprocedural complications were observed. 6 months after the procedure, control CT or MR angiography showed in almost all cases exclusion of the aneurysm from the circulation and normal blood flow in the treated artery. Neurological status six months after the procedure was normal in all patients. CONCLUSIONS Treatment of aneurysms with flow diverter Pipeline device is a safe and significantly less time consuming method in comparison with standard techniques. This new method is a promising approach in treatment of broad neck aneurysms.


title: Heterogeneous nuclear ribonucleoprotein a1 binds to the 3'-untranslated region and mediates potential 5'-3'-end cross talks of mouse hepatitis virus RNA.

text: The 3'-untranslated region (3'-UTR) of mouse hepatitis virus (MHV) RNA regulates the replication of and transcription from the viral RNA. Several host cell proteins have previously been shown to interact with this regulatory region. By immunoprecipitation of UV-cross-linked cellular proteins and in vitro binding of the recombinant protein, we have identified the major RNA-binding protein species as heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1). A strong hnRNP A1-binding site was located 90 to 170 nucleotides from the 3' end of MHV RNA, and a weak binding site was mapped at nucleotides 260 to 350 from the 3' end. These binding sites are complementary to the sites on the negative-strand RNA that bind another cellular protein, polypyrimidine tract-binding protein (PTB). Mutations that affect PTB binding to the negative strand of the 3'-UTR also inhibited hnRNP A1 binding on the positive strand, indicating a possible relationship between these two proteins. Defective-interfering RNAs containing a mutated hnRNP A1-binding site have reduced RNA transcription and replication activities. Furthermore, hnRNP A1 and PTB, both of which also bind to the complementary strands at the 5' end of MHV RNA, together mediate the formation of an RNP complex involving the 5'- and 3'-end fragments of MHV RNA in vitro. These studies suggest that hnRNP A1-PTB interactions provide a molecular mechanism for potential 5'-3' cross talks in MHV RNA, which may be important for RNA replication and transcription.


title: Bracing for the Big Ones: Novel Diseases like COVID-19 Could Test Texas' Public Health System.

text: Novel diseases like COVID-19 could be a tough test for Texas' public health system.


title: [Efficiency of the quarantine system during the epidemic of severe acute respiratory syndrome in Beijing, 2003].

text: OBJECTIVE An epidemic of severe acute respiratory syndrome (SARS) hit Beijing, China, between March and July 2003 with an attack rate of 1.9 per 10 thousand. (2,521 cases). To control the epidemic of SARS, a total of 30,173 residents were quarantined either in their residence or in quarantine sites. In order to understand the personal need of being quarantined and to estimate the risk of developing SARS during the quarantine period, a survey on the quarantined residents of Haidian District, Beijing, China was carried out. METHODS 33 precincts in Haidian District divided into five groups (7 in north, 6 in south, 7 in west, 6 in east and 7 in central of Haidian District) according to the location of the precincts were involved. The director of Center of Disease Control and Prevention of Haidian District was asked to select 1 precinct from each group according to the workload of the precinct quarantine officers. From those 5 precincts we obtained lists of all quarantined persons from the precinct quarantine officers. All quarantinees were asked to complete a self-administered questionnaire. The SARS patients were diagnosed and verified according to the diagnosis criteria released by Chinese Ministry of Health which was equivalent to the SARS 'probable case' definition of WHO. All SARS cases had been reported to the relative authorities. RESULTS By May 23, 2003, 5,186 persons had been quarantined in Haidian district, accounting for 0.23% of all residents. 1,028 of sampled quarantined residents completed the questionnaire. Of those who completed the questionnaire, 2.3% (95% CI: 1.5% - 3.5%) developed SARS while under quarantine. The median quarantine period was 14 days (range 1 day to 28 days). 61% of the quarantinees had a direct contact history with SARS patients, and all secondary SARS patients developed through contact to these quarantinees. The remaining 39% quarantinees who did not have a direct contact history with SARS patients had not developed SARS during the period under quarantine. 37% of the quarantees had direct contact during original patients' symptomatic period with an AR of 6.3%. Persons who looked after the illed SARS patient(s) during their symptomatic period, had an highest attack rate of 31% (95% CI: 20% - 44%). 63% (95% CI: 60% - 66%) of the total quarantined persons did not have direct contact with a SARS patient during his/her symptomatic period, with an attack rate of 0% (95% CI: 0% - 0.73%). CONCLUSION Only those persons having direct contact with ill SARS patients need to be under quarantine. The overall cost for quarantine on SARS prevention could be reduced by as much as 63% if the quarantine program was limited to this group. No evidence was found that SARS patient was infective during the incubation period.


title: Elevated plasma tissue-type plasminogen activator (t-PA) and soluble thrombomodulin in patients suffering from severe acute respiratory syndrome (SARS) as a possible index for prognosis and treatment strategy.

text: OBJECTIVE To detect the presence of endothelial injury in patients with severe acute respiratory syndrome (SARS) via enhanced levels of tissue-type plasminogen activator (t-PA) and soluble thrombomodulin (sTM). METHODS Case patients were from Xuanwu Hospital (Capital University of Medical Sciences, Beijing, China), and all of them met clinical criteria for SARS. Healthy controls were some of the hospital employees. Endothelial injury bio-markers tPA and sTM were detected by commercial ELISA-methods. RESULTS Classic plasma markers of endothelial injury, tPA and sTM significantly elevated in SARS patients in comparison to controls [t-PA: 1.48 +/- 0.16 nmol/L versus 0.25 +/- 0.03 nmol/L (P<0.0001), and sTM: 0.26 +/- 0.06 nmol/L versus 0.14 +/- 0.02 nmol/L (P<0.05)]. The only patient who died had extremely high levels of these endothelial injury markers (t-PA: 2.77 nmol/L and sTM: 1.01 nmol/L). The likelihood ratio analysis indicated the excellent discriminating power for SARS at the optimal cut-point of 0.49 nmol/L for tPA and 0.20 nmol/L for sTM, respectively. Significant numerical correlations were found among these endothelial injury markers in SARS patients. The numerical coefficient of correlation Pearson r between t-PA and sTM was 0.5867 (P<0.05). CONCLUSION Increased plasma concentrations of tPA and sTM in patients with SARS suggest the possibility of endothelial injury. SARS patients might need anticoagulant therapy or fibrinolytic therapy in order to reverse intraalveolar coagulation, microthrombi formation, alveolar and interstitial fibrin deposition. It may not only provide a useful treatment and prognostic index but also allow a further understanding of the pathological condition of the disease.


Is Australia prepared for the next pandemic?


docid: hdtwugxx

title: Changes to management of a non-pandemic illness during the COVID-19 pandemic: case study of invasive management of acute coronary syndrome.

text: The coronavirus 2019 (COVID-19) pandemic requires significant changes to standard operating procedures for non-COVID-19 related illnesses. Balancing the benefit from standard evidence-based treatments with the risks posed by COVID-19 to patients, healthcare workers and to the population at large is difficult due to incomplete and rapidly changing information. In this article, we use management of acute coronary syndromes as a case study to show how these competing risks and benefits can be resolved, albeit incompletely. While the risks due to COVID-19 in patients with acute coronary syndromes is unclear, the benefits of standard management are well established in this condition. As an aid to decision making, we recommend systematic estimation of the risks and benefits for management of any condition where there is likely to be an increase in non-COVID-19 related mortality and morbidity due to changes in routine care.


title: Selection of CTL escape mutants in mice infected with a neurotropic coronavirus: quantitative estimate of TCR diversity in the infected central nervous system.

text: Variant viruses mutated in the immunodominant cytotoxic T cell epitope surface (S) glycoprotein S-510-518 are selected in mice chronically infected with mouse hepatitis virus, strain JHM. We determined whether this selection occurred in the presence of an oligoclonal or polyclonal T cell response using soluble MHC/peptide tetramers in direct ex vivo analyses of CNS-derived lymphocytes. A total of 42% (range, 29-60%) of CD8 T cells in the CNS of mice with acute encephalitis recognized epitope S-510-518. A total of 34% (range, 18-62%) of cells from mice with hind limb paralysis (and chronic demyelination) were also epitope specific, even though only virus expressing mutated epitope is detected in these animals. Sequence analysis of the beta-chain CDR3 of 487 tetramer S-510-518-positive cDNA clones from nine mice showed that a majority of clonotypes were identified in more than one mouse. From these analyses, we estimated that 300-500 different CD8 T cell clonotypes responsive to epitope S-510-518 were present in each acutely infected brain, while 100-900 were present in the CNS of each mouse with chronic disease. In conclusion, a polyclonal CD8 T cell response to an epitope does not preclude the selection of T cell escape mutants, and epitope-specific T cells are still present at high levels even after RNA-encoding wild-type sequence is no longer detectable.


title: Feline coronavirus type II strains 79-1683 and 79-1146 originate from a double recombination between feline coronavirus type I and canine coronavirus.

text: Recent evidence suggests that the type II feline coronavirus (FCoV) strains 79-1146 and 79-1683 have arisen from a homologous RNA recombination event between FCoV type I and canine coronavirus (CCV). In both cases, the template switch apparently took place between the S and M genes, giving rise to recombinant viruses which encode a CCV-like S protein and the M, N, 7a, and 7b proteins of FCoV type I (K. Motowaka, T. Hohdatsu, H. Hashimoto, and H. Koyama, Microbiol. Immunol. 40:425-433, 1996; H. Vennema, A. Poland, K. Floyd Hawkins, and N. C. Pedersen, Feline Pract. 23:40-44, 1995). In the present study, we have looked for additional FCoV-CCV recombination sites. Four regions in the pol gene were selected for comparative sequence analysis of the type II FCoV strains 79-1683 and 79-1146, the type I FCoV strains TN406 and UCD1, the CCV strain K378, and the TGEV strain Purdue. Our data show that the type II FCoVs have arisen from double recombination events: additional crossover sites were mapped in the ORF1ab frameshifting region of strain 79-1683 and in the 5' half of ORF1b of strain 79-1146.


title: Screening and identification of linear B-cell epitopes and entry-blocking peptide of severe acute respiratory syndrome (SARS)-associated coronavirus using synthetic overlapping peptide library.

text: A 10-mer overlapping peptide library has been synthesized for screening and identification of linear B-cell epitopes of severe acute respiratory syndrome associated coronavirus (SARS-CoV), which spanned the major structural proteins of SARS-CoV. One hundred and eleven candidate peptides were positive according to the result of PEPscan, which were assembled into 22 longer peptides. Five of these peptides showed high cross-immunoreactivities (approximately 66.7 to 90.5%) to SARS convalescent patients' sera from the severest epidemic regions of the China mainland. Most interestingly, S(471-503), a peptide located at the receptor binding domain (RBD) of SARS-CoV, could specifically block the binding between the RBD and angiotensin-converting enzyme 2, resulting in the inhibition of SARS-CoV entrance into host cells in vitro. The study demonstrated that S(471-503) peptide was a potential immunoantigen for the development of peptide-based vaccine or a candidate for further drug evaluation against the SARS-CoV virus-cell fusion.


docid: ivqrmdof

title: A study of naturally occurring feline coronavirus infections in kittens.

text: Feline coronavirus is a common infection in cats, as indicated by the high prevalence of antibodies against the virus, especially in multicat households. Approximately 5 to 12 per cent of seropositive cats develop classical feline infectious peritonitis. A survey of kittens born into households of seropositive cats demonstrated the existence of healthy coronavirus carriers. Seronegative animals did not appear to excrete virus. No specific antibody titre could be linked to carrier status and some carrier cats subsequently became seronegative. The management of the kittens strongly influenced whether they became infected, and some degree of protection appeared to be conferred by maternally derived antibody. At present, feline infectious peritonitis virus and feline enteric coronavirus can only be differentiated by their different clinical histories in infected catteries. In this survey, cases of feline infectious peritonitis occurred in kittens from households where the initial presentation had been enteritis and vice versa. Therefore no difference in epidemiology could be found.


docid: synpynwo

title: Virology and immunology of the common cold.

text: The common cold is a complex infectious syndrome caused by any one of a large number of antigenitically distinct viruses found in four groups. These groups are the myxo- and paramyxoviruses, the adenoviruses, the rhinoviruses and the coronaviruses. The members of the different groups differ in their physical, biochemical, and immunologic characteristics. With currently available methods, it is possible to determine the cause of 60-70% of colds. The large rhinovirus group is the most important of the known common cold viruses, accounting for approximately 30% of colds. These small RNA viruses have a genome of 7000 nucleotides, which shares considerable homology with poliovirus. The capsid of the rhinovirus is loosely packed, resulting in a relative acid sensitivity compared to the enteroviruses. Although there are at least 89 different antigenic types, all rhinoviruses attach to either one of two cellular receptors. Immunity to rhinovirus is type-specific and associated with neutralizing antibody in nasal secretions and serum. There is a steady acquisition of antibody to the rhinoviruses during childhood and adolescence. The rhinoviruses may be undergoing slow antigenic drift.


title: Are there characteristics of infectious diseases that raise special ethical issues?

text: This paper examines the characteristics of infectious diseases that raise special medical and social ethical issues, and explores ways of integrating both current bioethical and classical public health ethics concerns. Many of the ethical issues raised by infectious diseases are related to these diseases' powerful ability to engender fear in individuals and panic in populations. We address the association of some infectious diseases with high morbidity and mortality rates, the sense that infectious diseases are caused by invasion or attack on humans by foreign micro-organisms, the acute onset and rapid course of many infectious diseases, and, in particular, the communicability of infectious diseases. The individual fear and community panic associated with infectious diseases often leads to rapid, emotionally driven decision making about public health policies needed to protect the community that may be in conflict with current bioethical principles regarding the care of individual patients. The discussion includes recent examples where dialogue between public health practitioners and medical-ethicists has helped resolve ethical issues that require us to consider the infected patient as both a victim with individual needs and rights and as a potential vector of disease that is of concern to the community.


title: Infectious diseases and governance of global risks through public communication and participation.

text: In recent years a succession of health emergencies connected with the threat of new, possibly global, infectious diseases has stimulated the attention of the mass media, the scientific community, and international public opinion, setting a tough test for the institutions whose job is to manage the risks. On the basis of experience in the fields of AIDS, BSE, SARS and bird flu, this study discusses the strong and weak points of governance procedures for health risks. In particular, the paper illustrates how risk management can be improved by adopting practices and procedures which actively involve the public in dealing with the emergency, by taking a transparent and accessible approach to communication with the public (including the provision of information about the risks) and by fostering the unrestricted exchange of scientific knowledge among researchers. Lastly, the text shows how the analysis of these themes provides starting points for understanding the crisis in the current relationship between science and society.


title: [Recommendations of antimicrobial treatment in patients allergic to beta-lactam antibiotics].

text: Beta-lactam antibiotics are the cornerstone of most of the severe bacterial infections. However, their use can be limited by resistances and allergic reactions. Allergic reactions to beta-lactam antibiotics account for only a small proportion of reported adverse drug reactions, but they are related with an important morbidity, mortality and increase of the health care costs. Drug-specific IgE antibodies cause early reactions, whereas T cells play a predominant role in delayed hypersensitivity reactions. For penicillin a major antigenic determinant and several minor determinants have been identified. Clinical assessment is mandatory by medical history, skin and other testing, including provocation. If the beta-lactam should be avoided or a desensitization procedure should be performed depends on the nature and severity of the reaction. Several new antibiotics are currently available (tigecycline, linezolid, daptomycin, etc.) that are as effective and safe as beta-lactams. In this article we have developed a few recommendations for the management of patients with allergy to beta-lactams on the basis of evidence and expert opinion.


docid: lsorlpcr

title: Implementation of a quality systems approach for laboratory practice in resource-constrained countries.

text: Under the direction of the US Global AIDS Coordinator's Office, Department of Health and Human Services, the CDC Global AIDS programme helps resource-constrained countries to address the global HIV/AIDS pandemic. Activities include laboratory capacity and laboratory infrastructure development in 25 resource-constrained countries. Medical practitioners and public health programme leaders in industrialized countries rely on the use of quality laboratory data for evidence-based medical decision-making to determine policy for the implementation of disease control measures, to monitor disease to determine the impact of control programmes, and to support surveillance activities. In these countries, laboratory data to support decision-making processes have a level of quality attributable to laws, regulations and guidelines developed over many years. However, resource-constrained countries have not had similar experiences. Few countries have developed laws, regulations or guidelines, nor is there a data-use culture (e.g. evidence-based medicine) for those in the decision-making environment in resource-constrained countries. The strengthening of laboratory capability and capacity in resource-constrained countries is an important goal to improve accurate and reliable data for the diagnosis, treatment and monitoring of disease.A process for the implementation of a quality systems approach for a laboratory is presented: (i) acknowledgement of the need to improve the laboratory programme in the country at the Ministry of Health and at all decision-making levels within the provinces/states of the country; (ii) assessment of capabilities, capacities, infrastructure, and training needs; (iii) implementation of a national meeting of laboratorians; (iv) designation of a national Quality Assurance Office and leadership within that office; (v) the development and provision of technical training.


title: Is It Rational to Perform Liver Resection for Patients with Intermediate and Advanced Stages of Hepatocellular Carcinoma?

text: This study aimed to investigate clinical characteristics of hepatocellular carcinoma and the outcome of our aggressive treatment policy which follows the Barcelona Clinic Liver Cancer (BCLC) guidance. In this study, we retrospectively analyzed data of 102 patients who were treated for hepatocellular carcinoma between January 2007 and October 2016. Male predominance (81.4%) and a median age of 61 years were observed. Cirrhosis was evident in 88.2 per cent of patients. Viral hepatitis (77.5%) was the most common underlying etiology. The majority of our patients (71.6%) were in BCLC B and C stages. Liver resection was performed in 53.4 per cent of patients in those stages. Transarterial chemoembolization was the leading interventional treatment. Overall survival rates at three and five years were 75 per cent and 75 per cent in BCLC 0, 69 per cent and 58 per cent in BCLC A, 50 per cent and 41 per cent in BCLC B, and 11 per cent and 11 per cent in BCLC C, respectively. The BCLC treatment algorithm should consider the role of liver resection also for intermediate stages.


title: Effects of different antidepressant treatments on the core of depression.

text: Core symptoms of depression are a combination of psychological and somatic symptoms, often combined with psychomotor and cognitive disturbances. Diagnostic classification of depression including the concepts of melancholic, endogenous, or severe depression describe severely depressed patients suffering from most of the core symptoms, together with clinical characteristics of a cyclic unipolar or bipolar course, lower placebo response rates, higher response rates to electroconvulsive therapy, to antidepressant treatments with dually or mixed modes of action, or to lithium augmentation. Higher rates of hypothalamic-pituitary-adrenal axis hyperactivity and specific electroencephalographic patterns have also been shown in this patient group. Summarizing the symptomatology of depression in these patients, a broad overlap between the abovementioned subgroups can be suggested. Because the positive diagnosis of those core symptoms of depression may include clinical consequences, it would be of use to integrate all the mentioned concepts in the upcoming new versions of the diagnostic systems DSM-V and ICD-11.


title: Treatment of malignant lymphoma in an African lion (Panthera leo).

text: A 14 yr-old male, vasectomized African lion (Panthera leo) exhibited mild weight loss despite adequate appetite. Splenomegaly was diagnosed on physical examination. On the basis of hematology and clinical pathology, malignant lymphoma with chronic lymphocytic leukemia was diagnosed. Abdominal exploratory surgery and splenectomy were performed. Histologic examination and immunohistochemistry confirmed a small cell peripheral T-cell lymphoma. Initial treatments consisted of doxorubicin and prednisone, with later addition of lomustine. The lion remained in clinical remission at 2 mo, 6 mo, and 12 mo postchemotherapy physical examinations. The lion survived 504 days from initial diagnosis. At necropsy, the only lesions consistent with lymphoma were localized epitheliotrophic infiltrates of small neoplastic T lymphocytes within the nasopharyngeal epithelium and the underlying submucosa observed on microscopic examination.


docid: uoqkfjie

title: Severe acute respiratory syndrome coronavirus group-specific open reading frames encode nonessential functions for replication in cell cultures and mice.

text: SARS coronavirus (SARS-CoV) encodes several unique group-specific open reading frames (ORFs) relative to other known coronaviruses. To determine the significance of the SARS-CoV group-specific ORFs in virus replication in vitro and in mice, we systematically deleted five of the eight group-specific ORFs, ORF3a, OF3b, ORF6, ORF7a, and ORF7b, and characterized recombinant virus replication and gene expression in vitro. Deletion of the group-specific ORFs of SARS-CoV, either alone or in various combinations, did not dramatically influence replication efficiency in cell culture or in the levels of viral RNA synthesis. The greatest reduction in virus growth was noted following ORF3a deletion. SARS-CoV spike (S) glycoprotein does not encode a rough endoplasmic reticulum (rER)/Golgi retention signal, and it has been suggested that ORF3a interacts with and targets S glycoprotein retention in the rER/Golgi apparatus. Deletion of ORF3a did not alter subcellular localization of the S glycoprotein from distinct punctuate localization in the rER/Golgi apparatus. These data suggest that ORF3a plays little role in the targeting of S localization in the rER/Golgi apparatus. In addition, insertion of the 29-bp deletion fusing ORF8a/b into the single ORF8, noted in early-stage SARS-CoV human and civet cat isolates, had little if any impact on in vitro growth or RNA synthesis. All recombinant viruses replicated to wild-type levels in the murine model, suggesting that either the group-specific ORFs play little role in in vivo replication efficiency or that the mouse model is not of sufficient quality for discerning the role of the group-specific ORFs in disease origin and development.


Multiple congenital anomalies in a calf.


title: Luxury at a cost? Recombinant mouse hepatitis viruses expressing the accessory hemagglutinin esterase protein display reduced fitness in vitro.

text: Group 2 coronaviruses encode an accessory envelope glycoprotein species, the hemagglutinin esterase (HE), which possesses sialate-O-acetylesterase activity and which, presumably, promotes virus spread and entry in vivo by facilitating reversible virion attachment to O-acetylated sialic acids. While HE may provide a strong selective advantage during natural infection, many laboratory strains of mouse hepatitis virus (MHV) fail to produce the protein. Apparently, their HE genes were inactivated during cell culture adaptation. For this report, we have studied the molecular basis of this phenomenon. By using targeted RNA recombination, we generated isogenic recombinant MHVs which differ exclusively in their expression of HE and produce either the wild-type protein (HE+), an enzymatically inactive HE protein (HE0), or no HE at all. HE expression or the lack thereof did not lead to gross differences in in vitro growth properties. Yet the expression of HE was rapidly lost during serial cell culture passaging. Competition experiments with mixed infections revealed that this was not due to the enzymatic activity: MHVs expressing HE+ or HE0 propagated with equal efficiencies. During the propagation of recombinant MHV-HE+, two types of spontaneous mutants accumulated. One produced an anchorless HE, while the other had a Gly-to-Trp substitution at the predicted C-terminal residue of the HE signal peptide. Neither mutant incorporated HE into virion particles, suggesting that wild-type HE reduces the in vitro propagation efficiency, either at the assembly stage or at a postassembly level. Our findings demonstrate that the expression of "luxury" proteins may come at a fitness penalty. Apparently, under natural conditions the costs of maintaining HE are outweighed by the benefits.


BK virus vireamia and viruria among a group of post kidney transplant patients in Sri Lanka


title: trans-encapsidation of a poliovirus replicon by different picornavirus capsid proteins.

text: A trans-encapsidation assay was established to study the specificity of picornavirus RNA encapsidation. A poliovirus replicon with the luciferase gene replacing the capsid protein-coding region was coexpressed in transfected HeLa cells with capsid proteins from homologous or heterologous virus. Successful trans-encapsidation resulted in assembly and production of virions whose replication, upon subsequent infection of HeLa cells, was accompanied by expression of luciferase activity. The amount of luciferase activity was proportional to the amount of trans-encapsidated virus produced from the cotransfection. When poliovirus capsid proteins were supplied in trans, >2 x 10(6) infectious particles/ml were produced. When coxsackievirus B3, human rhinovirus 14, mengovirus, or hepatitis A virus (HAV) capsid proteins were supplied in trans, all but HAV showed some encapsidation of the replicon. The overall encapsidation efficiency of the replicon RNA by heterologous capsid proteins was significantly lower than when poliovirus capsid was used. trans-encapsidated particles could be completely neutralized with specific antisera against each of the donor virus capsids. The results indicate that encapsidation is regulated by specific viral nucleic acid and protein sequences.


docid: vessmdue

title: The biology of NKT cells.

text: Recognized more than a decade ago, NKT cells differentiate from mainstream thymic precursors through instructive signals emanating during TCR engagement by CD1d-expressing cortical thymocytes. Their semi-invariant alphabeta TCRs recognize isoglobotrihexosylceramide, a mammalian glycosphingolipid, as well as microbial alpha-glycuronylceramides found in the cell wall of Gram-negative, lipopolysaccharide-negative bacteria. This dual recognition of self and microbial ligands underlies innate-like antimicrobial functions mediated by CD40L induction and massive Th1 and Th2 cytokine and chemokine release. Through reciprocal activation of NKT cells and dendritic cells, synthetic NKT ligands constitute promising new vaccine adjuvants. NKT cells also regulate a range of immunopathological conditions, but the mechanisms and the ligands involved remain unknown. NKT cell biology has emerged as a new field of research at the frontier between innate and adaptive immunity, providing a powerful model to study fundamental aspects of the cell and structural biology of glycolipid trafficking, processing, and recognition.


title: Proprotein convertases: lessons from knockouts.

text: The physiological role of the subtilisin/kexin-like proprotein convertases (PCs) in rodents has been examined through the use of knockout mice. This review will summarize the major in vivo defects that result from the disruption of the expression of their genes. This includes abnormal embryonic development, hormonal disorder, infertility, and/or modified lipid/sterol metabolism. Members of the PC family play a central role in the processing of various protein precursors ranging from hormones and growth factors to bacterial toxins and viral glycoproteins. Proteolysis occurring at basic residues is mediated by the basic amino acid-specific proprotein convertases, namely: PC1/3, PC2, furin, PACE4, PC4, PC5/6, and PC7. In contrast, proteolysis at nonbasic residues is performed by the subtilisin/kexin-like isozyme-1 (SKI-1/S1P) and the newly identified neural apoptosis-regulated convertase-1 (PCSK9/NARC-1). In addition to their requirement for many physiological processes, these enzymes are also involved in various pathologies such as cancer, obesity, diabetes, lipid disorders, infectious diseases, atherosclerosis and neurodegenerative diseases.


The Problems of Virology.