title: Tropism of human adenovirus type 5-based vectors in swine and their ability to protect against transmissible gastroenteritis coronavirus.

text: The infection of epithelia] swine testicle and intestinal porcine epithelial (IPEC-1) cell lines by adenovirus type 5 (Ad5) has been studied in vitro by using an Ad5-luciferase recombinant containing the firefly luciferase gene as a reporter. Porcine cell lines supported Ad5 replication, showing virus titers, kinetics of virus production, and luciferase expression levels similar to those obtained in human 293 cells, which constitutively express the 5'-end 11% of the Ad5 genome. The tropism of Ad5-based vectors in swine and its ability to induce an efficient immune response against heterologous antigens expressed by foreign genes inserted in these vectors has been determined. Ad5 vectors replicate and express heterologous antigens in porcine lungs and mediastinal and mesenteric lymph nodes. Significant levels of heterologous antigen expression were also demonstrated in the small intestine (jejunum and ileum), but Ad5 replication in this organ was very poor, suggesting that Ad vectors undergo an abortive replication in the porcine small intestine. The tissues infected by Ad5 were dependent on the inoculation route. The oronasal route appeared to be best for inoculation of bronchus-associated lymphoid tissue infection, while the intraperitoneal route was best for gut-associated lymphoid tissue infection. Epithelial cells of bronchioles, macrophages, type II pneumocytes, and follicular dendritic cells were identified as targets for Ad5, while epithelial cells of the intestine were not infected by Ad5. Viruses with a deletion from 79.5 to 84.8 map units in the E3 region, with or without heterologous inserted genes, replicated to lower levels in porcine tissues than did wild-type Ad5. It was also shown that an Ad5 recombinant expressing the four antigenic sites (A, B, C, and D) of transmissible gastroenteritis coronavirus (TGEV) spike protein induced in swine immune responses which neutralized TGEV infectivity. In addition, porcine serum from Ad-TGEV-immune animals provide passive protection when mixed with fully virulent TGEV and orally administered to highly susceptible newborn piglets. These results taken together indicate that swine may be a good animal model for human Ad5 lung infection to aid in the evaluation of candidate adenovirus vaccines and that Ad5 may be suitable as a recombinant viral vaccine or for other applications in swine.


title: [Analysis of medication characteristics of traditional Chinese medicine in treating coronavirus disease-19 based on data mining].

text: OBJECTIVE To analysis the medication characteristics of the prescriptions issued via open channel by the National and Provincial Health Committee and the State Administration of Traditional Chinese Medicine in treating coronavirus disease 2019 (COVID-19). METHODS We collected the data of traditional Chinese medicine related to treatment plans published by the National and Provincial Health Committee and the State Administration of Traditional Chinese Medicine from the start of COVID-19 outbreak in Wuhan to February 19, 2020. The frequency analysis, cluster analysis and correlation analysis were performed. RESULTS The study collected 4 national and 34 regional prevention and treatment plans, 578 items, 84 traditional Chinese formulations, 60 Chinese patent medicines, and 230 Chinese herbs. The high frequently used herbs were Liquorice, Scutellariabaicalensis, Semen armeniacaeamarae, and Gypsum. The commonly used traditional formulations included Maxing Shigan Decoction, Yin Qiao Powder, and Xuanbai Chengqi Decoction. The Chinese patent drugs included Angong Niuhuang Pill, Xuebijing Injection, and Lianhua Qingwen Capsule. The most common paired medications were Ephedra and Semen armeniacaeamarae, Fructusforsythiae and Liquorice. Two core combinations and one novel formula were discovered in the study. CONCLUSIONS Yin Qiao Powder and Huopo Xialing Decoction are the basic formulations for Weifen syndrome of COVID-19. In addition, Maxing Shigan Decoction, Liang Ge Powder, Qingwen Baidu Decoction and Da Yuan Decoction are the basic formulations for Qifen syndrome of COVID-19. The main medication characteristics are clearing heat, entilating lung, removing toxicity and removing turbidity. It shows that removing toxicity and eliminating evil are the prescription thought in treating epidemic disease of traditional Chinese medicine.


title: SARS: clinical presentation, transmission, pathogenesis and treatment options.

text: SARS (severe acute respiratory syndrome) appeared as the first emerging infectious disease of this century. It is fortunate that the culprit virus can be grown without much difficulty from a commonly used cell line, allowing an unlimited supply of isolates for further molecular studies and leading to the development of sensitive diagnostic assays. How the virus has successfully jumped the species barrier is still a mystery. The superspreading events that occurred within hospital, hotel and high-density housing estate opens a new chapter in the mechanisms and routes of virus transmission. The old practice of quarantine proved to be still useful in controlling the global outbreak. Despite all the available sophisticated tests, alertness with early recognition by healthcare workers and prompt isolation of suspected cases is still the most important step for containing the spread of the infection. Although the rapidly evolving outbreak did not allow the conducting of systematic clinical trails to evaluate treatment options, the accumulated experience on managing SARS patients will improve the clinical outcome should SARS return. Although SARS led to more than 700 deaths worldwide, the lessons learnt have prepared healthcare systems worldwide to face future emerging and re-emerging infections.


title: The organisation of primary care in Europe: Part 2 Agenda--position paper of the European Forum for Primary Care.

text: The contemporary models and trends of European organisational developments in primary care, identified in part 1 of this article, are the subject of analysis and discussion. Four main issues are identified in relation to the future protection and progress of primary care, and a series of policy interventions specified. These are directed at international agencies and action. Two new case study summaries are supplied as illustrations of the dilemmas now being encountered by primary care organisations across the extended Europe. With some supplementary material, the article is an edited version of the 2008 European Forum for Primary Care (EFPC) position paper on the organisation of primary care in Europe.


title: Ulcerative duodenitis in foals.

text: Seven foals aged 18 days to 3 1/2 months had either single or multiple full-circumference segments or long antimesenteric bands of necrotizing duodenitis, sharply delineated from adjacent viable duodenum. Perforation of the necrotic wall had occurred in all foals, leading to acute fibrinous peritonitis. On the mucosal surface severe diffuse, acute inflammation and ulceration involved the anterior half of the duodenum. Two further foals, aged 28 and 30 days, had lesions that are believed to be a chronic form of this disease. Both foals had a thickened duodenal wall, with large areas of mucosa replaced by granulation tissue. In addition one had several strictures associated with firm adhesions between the duodenal serosa and adjacent structures, together with ascending cholangiohepatitis and pancreatitis. Eight foals had gastric ulcers that were considered to be of less significance than the duodenal lesions. No etiologic agent could be found by aerobic or anaerobic bacterial culturing, negative contrast electron microscopy for viruses, or immunofluorescence staining for equine herpesvirus 1, equine adenovirus, or equine coronavirus. The possible involvement of non-steroidal anti-inflammatory drugs is discussed.


title: [Determination of SARS-CoV by simplified nested fluorescent RT-PCR].

text: OBJECTIVE To establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR. METHODS Primers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested. RESULTS The positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160). CONCLUSION The simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.


Cryptosporidiosis in a pup with distemper.


Biologics and Small Molecules in the Treatment of COVID-19.


title: Genetic predisposition to coronavirus-induced retinal disease.

text: PURPOSE Retinal inflammatory and degenerative processes in humans and animals frequently are associated with genetic factors. The murine coronavirus, mouse hepatitis virus (MHV), JHM strain, induces a biphasic retinal disease in adult BALB/c mice. The genetic constitution of the host and the virus serotype can be critical factors in determining the outcome of a virus infection. The purpose of this study was to evaluate the possible role of host genetics in murine coronavirus-induced retinal disease. METHODS JHM virus was inoculated by the intravitreal route into BALB/c, CD-1, and A/J mice. At varying times after inoculation, eye tissues were evaluated histologically. Antibody responses to the virus were evaluated by neutralization assays. RESULTS JHM virus induces a biphasic retinal disease in BALB/c mice. In the early phase, 1 to 7 days after inoculation, retinal vasculitis is observed. The second phase, characterized by retinal degeneration in the absence of inflammation, is seen by day 10 and progresses for several months. There is a similar biphasic disease process in JHM virus-infected A/J mice. However, retinal changes are less severe than those seen in BALB/c mice. Retinal tissue damage induced by JHM virus in CD-1 mice is different. Only the early phase of the disease, consisting of retinal vasculitis, was observed. These CD-1 mice do not develop the retinal degenerative disease. In fact, after day 10, the retina has a normal appearance. These differences in retinal tissue damage are seen over a wide range of infectivity of the virus inocula. Virus concentrations ranging from 10(1.4) to 10(4.4) TCID50/5 microliters were capable of inducing both inflammation and degeneration in BALB/c mice, whereas, the highest concentration of virus (10(4.4) TCID50/5 microliters) in CD-1 mice resulted in only the early inflammatory changes. CONCLUSIONS The authors show that the genetics of the host can profoundly affect the nature of retinal tissue damage. These studies substantiate the concept that a virus can indeed trigger retinal degenerative processes in genetically susceptible hosts.


Requirements for memory maintenance.


Maculopathy in dengue fever.


HIV: A journey just begun.


title: Is Laparoendoscopic Single-Site Adrenalectomy a Feasible Alternative in Treating Aldosterone-Producing Adenoma?

text: Objective. To compare laparoendoscopic single-site (LESS) and conventional multiport adrenalectomy in patients with aldosterone-producing adenoma (APA). Material and Methods. We retrospectively reviewed patients who had been clinically confirmed with unilateral APA and who underwent LESS or multiport adrenalectomy between 2009 and 2014. Perioperative data were obtained for all patients. Blood pressure and the levels of serum aldosterone, renin, and potassium were checked periodically. Results. We identified 45 APA patients in the LESS group and 71 in the multiport group. The baseline characteristics were matched between two groups. All adrenalectomies were completed successfully, except one with laparoscopic conversion in the single-port group and one open conversion in the multiport group. After a mean follow-up around one year, there were no significant group differences in the improvement of hypertension, number of types of medication taken, and cure of hypokalemia after operation. Conclusions. Our study confirm that LESS adrenalectomy achieved similar clinical and functional outcomes as conventional multiport adrenalectomy for management of unilateral APA.


title: [Ligand screening in affinity chromatography and its applications].

text: Affinity chromatography is one of the most powerful techniques in selective purification and isolation of a great number of compounds. Affinity ligand is the most important factor in affinity chromatography, therefore, the selection and the screening of affinity ligand become the first issue to be considered when a novel type of affinity packing or a novel system of affinity chromatography needs to be developed. In combination with the research work of the author' s group, this paper briefly summaries the recent progresses in selection, screening and application of affinity ligands. A new screening method with antisense peptide based combinatorial peptide libraries has been developed to search the affinity ligands for target proteins and peptides. This strategy is simpler and more effective than the conventional combinatorial strategy. The work including syntheses and screening is greatly reduced and the structural determination of the ligands becomes easy by using this method. Furthermore, the new method has been successfully applied to the screening of inhibitors for influenza A virus and SARS coronavirus, as well as the construction of quartz crystal microbalance ( QCM) biosensor for human interferon-beta.


Biochemistry of coronaviruses 1980.


title: Current global vaccine and drug efforts against COVID-19: Pros and cons of bypassing animal trials.

text: COVID-19 has become one of the biggest health concern, along with huge economic burden. With no clear remedies to treat the disease, doctors are repurposing drugs like chloroquine and remdesivir to treat COVID-19 patients. In parallel, research institutes in collaboration with biotech companies have identified strategies to use viral proteins as vaccine candidates for COVID-19. Although this looks promising, they still need to pass the test of challenge studies in animal models. As various models for SARS-CoV-2 are under testing phase, biotech companies have bypassed animal studies and moved to Phase I clinical trials. In view of the present outbreak, this looks a justified approach, but the problem is that in the absence of animal studies, we can never predict the outcomes in humans. Since animal models are critical for vaccine development and SARS-CoV-2 has different transmission dynamics, in this review we compare different animal models of SARS-CoV-2 with humans for their pathogenic, immune response and transmission dynamics that make them ideal models for vaccine testing for COVID-19. Another issue of using animal model is the ethics of using animals for research; thus, we also discuss the pros and cons of using animals for vaccine development studies.


title: Single-port laparoscopic surgery by use of a surgical glove port: initial experience with 25 cases.

text: BACKGROUND single-port laparoscopic surgery has gained popularity over the last decade. This technique is used for several surgical procedures. This paper documents a new and cheaper access method known as "surgical glove port" or "homemade single-port", and describes our initial experience with 25 cases. METHOD Eleven cholecystectomies, eight totally extraperitoneal inguinal hernia repairs, three splenectomies, two gastric wedge resections, and one anti-reflux procedure were performed. RESULTS Twenty-four procedures out of twenty-five were completed without conversion to open or multiple port techniques. An individual patient who had both cholelithiasis and gastrointestinal stromal tumor underwent both cholecystectomy and gastric wedge resection in the same session with surgical glove port technique. In another case two additional 5-mm incisions were made for hemostasis. The additional operation was required and performed by using surgical glove port for a hiatal hernia repair case, because of postoperative hemorrhage. CONCLUSION Single-port laparoscopic surgery is an applicable technique for plenty of surgical procedures. Also, surgical glove port is a newer technique and some initial experience shave already published all over the world. Surgical glove port has advantages such as cost-effectiveness, easy and safe port implantation and specimen extraction. SGP can also be used for treating post-operative complications.


title: A nucleolar TAR decoy inhibitor of HIV-1 replication.

text: Tat is a critical regulatory factor in HIV-1 gene expression. It mediates the transactivation of transcription from the HIV-1 LTR by binding to the transactivation response (TAR) element in a complex with cyclin T1. Because of its critical and early role in HIV gene expression, Tat and its interaction with the TAR element constitute important therapeutic targets for the treatment of HIV-1 infection. Based on the known nucleolar localization properties of Tat, we constructed a chimeric small nucleolar RNA-TAR decoy that localizes to the nucleoli of human cells and colocalizes in the nucleolus with a Tat-enhanced GFP fusion protein. When the chimeric RNA was stably expressed in human T lymphoblastoid CEM cells it potently inhibited HIV-1 replication. These results demonstrate that the nucleolar trafficking of Tat is critical for HIV-1 replication and suggests a role for the nucleolus in HIV-1 viral replication.


title: Comparison of the inhibitory effects of interferon alfacon-1 and ribavirin on yellow fever virus infection in a hamster model.

text: Antiviral compounds were evaluated for efficacy against yellow fever virus (YFV) in a hamster model of YFV-induced liver disease. Challenge with a 10(2) 50% cell culture infectious doses of YFV resulted in a 50-80% mortality rate in female hamsters. Virus was detected by quantitative real-time RT-PCR (QRT-PCR) in liver, kidney, spleen and serum with peak titers on 4-6 days post-viral challenge (dpi). Serum levels of alkaline phosphatase, alanine aminotransferase (ALT), bilirubin, blood urea nitrogen, potassium and creatinine were significantly elevated, while serum levels of albumin, amylase, glucose, calcium, globulin, phosphorus, sodium and total protein were significantly reduced. Packed cell volume and white blood cell count were significantly elevated during the course of the infection. Intraperitoneal treatment of hamsters with 0.5-5 microg/kg/day interferon (IFN) alfacon-1, 100mg/kg/day viramidine or 50 mg/kg/day ribavirin, initiated 4h prior to YFV challenge, resulted in significant improvement in survival and serum ALT levels. Treatment with IFN alfacon-1 or ribavirin starting 2dpi, also significantly improved survival and serum ALT levels in hamsters challenged with YFV. Pre- and post-virus exposure treatment with IFN alfacon-1 was efficacious in improving disease in YFV-infected hamsters.


title: Vaccines against Francisella tularensis--past, present and future.

text: Francisella tularensis is a facultative intracellular bacterial pathogen capable of causing a spectrum of human diseases collectively called tularemia. The pathogen is highly infectious and some strains can cause rapidly lethal infection especially when inhaled. The latter were developed as biological weapons in the past and nowadays cause concern as potential bioterrorism agents. A live attenuated strain of the pathogen was developed more that 40 years ago and remains the sole prophylactic measure against the pathogen. Research to develop better live and subunit vaccines is under way. The former will require an understanding of the virulence factors of F. tularensis and a facile means of mutating them and the latter will require identification of the protective antigens of the pathogen. The current vaccine and its potential replacements are the focus of this review.


Pregnancy and Sars-Cov-2: A Novel Virus in a Unique Population


title: How often do you wash your hands? A review of studies of hand-washing practices in the community during and after the SARS outbreak in 2003.

text: We reviewed evidence of hand-washing compliance in community settings during the 2003 SARS outbreak. Literature was searched through PubMed, Cochrane Library, Wan Fang database and Google. English and Chinese papers were reviewed. Studies containing data on hand-washing, self-reported or directly observed, in community settings were selected. Case-control studies and studies in healthcare settings were excluded. Fourteen studies were reviewed. Self-reported hand-washing compliance increased in the first phase of the SARS outbreak and maintained a high level 22 months after the outbreak. The decline of hand-washing in Hong Kong after SARS was relatively slow. A significant gender difference in hand-washing compliance (female > male) was found in eight studies. The importance of family support and 'significant female others' in hand hygiene promotion are noted. The impact of education is uncertain. Perceived susceptibility to and severity of SARS, and perceived efficacy of hand-washing in preventing SARS, also predicted self-reported hand-washing compliance.


title: Neurovirological methods and their applications.

text: Over the last 30 years neurovirology has emerged as a major discipline which has much relevance to both human disease and many aspects of neuroscience. This overview of the field aims to define briefly most of the major neurovirological techniques, both "classical" and more recent, and to indicate how these have been used to gain knowledge about the pathogenesis, clinical investigation, and treatment of viral infections of the central nervous system.


title: Site-directed mutagenesis of the genome of mouse hepatitis virus by targeted RNA recombination.

text: We have genetically characterized a nucleocapsid (N) protein mutant of the coronavirus mouse hepatitis virus (MHV). This mutant, designated Alb4, is both temperature-sensitive and thermolabile, and its N protein is smaller than wild-type N. Sequence analysis of the Alb4 N gene revealed that it contains an internal deletion of 87 nucleotides, producing an in-frame deletion of 29 amino acids. All of these properties of Alb4 made it ideal for use as a recipient in a targeted RNA recombination experiment in which the deletion in Alb4 was repaired by recombination with synthetic RNA7, the smallest MHV subgenomic mRNA. Progeny from a cotransfection of Alb4 genomic RNA and synthetic RNA7 were selected for thermal stability. PCR analysis of candidate recombinants showed that they had regained the material that is deleted in the Alb4 mutant. They also had acquired a five nucleotide insertion in the 3' untranslated region, which had been incorporated into the synthetic RNA7 as a molecular tag. The presence of the tag was directly verified, as well, by sequencing the genomic RNA of purified recombinant viruses. This provided a clear genetic proof that the Alb4 phenotype was due to the observed deletion in the N gene. In addition, these results demonstrated that it is possible to obtain stable, independently replicating progeny from recombination between coronaviral genomic RNA and a tailored, synthetic RNA species. To date, we have constructed three additional mutants by this procedure. For two of these, a second-site point mutation that reverts the Alb4 phenotype has been transduced into a wild type background, which does not contain the Alb4 deletion.(ABSTRACT TRUNCATED AT 250 WORDS)


title: Lactic acid and glucose metabolism in healthy, lactic acid-infused, and diarrheic calves.

text: Kinetics of glucose and lactate metabolism were measured in healthy (normal) calves, healthy calves infused with lactate, and in calves which had received an enteropathogenic coronavirus inoculum and had become diarrheic. Lactate utilization was positively correlated with blood lactate concentration in the healthy calves, but its turnover was markedly reduced in diarrheic animals. The decreased turnover was not due to inhibition of lactate oxidation, but appeared to relate to decreased gluconeogenesis. Glucose entry rate was not affected by diarrhea, but was a function of glucose concentration.


title: HFE gene mutations in patients with acute leukemia.

text: An increased incidence of HFE gene mutations has been described in hematologic malignancies. In the present study, we investigated the allelic frequency of HFE gene mutations in 154 adult patients with acute leukemia (AL) [107 acute myeloid leukemia (AML), 20 acute promyelocytic leukemia (APL) and 27 acute lymphoblastic leukemia (ALL)]. The allelic frequency of the H63D mutation was 29% in AL patients and 25% in the healthy controls [P = 0.41; odds ratio (OR) = 1.20; 95% confidence interval (CI) = 0.77 - 1.93]. No difference was found between controls and AML or APL patients, whereas the H63D mutation was significantly more frequent in ALL than controls (44% vs. 25%, P = 0.04; OR = 2.37; 95% CI = 1.05 - 5.36). The overall comparison of the mutation among the three subtypes of AL demonstrated a higher allelic frequency in ALL (P = 0.02). In conclusion, our data demonstrate a correlation between the presence of the H63D mutation and the occurrence of ALL in adult patients.


title: 18F-2-deoxy-2-fluoro-D-glucose positron emission tomography, computed tomography, and magnetic resonance imaging for the detection of experimental colorectal liver metastases.

text: During the treatment of colorectal liver metastases, evaluation of treatment efficacy is of the utmost importance for decision making. The aim of the present study was to explore the ability of preclinical imaging modalities to detect experimental liver metastases. Nine male Wag/Rij rats underwent a laparotomy with intraportal injection of CC531 tumor cells. On days 7, 10, and 14 after tumor induction, sequential positron emission tomography (PET), computed tomography (CT), and magnetic resonance imaging (MRI) scans were acquired of each rat. At each time point, three rats were euthanized and the metastases in the liver were documented histologically. Topographically, the liver was divided into eight segments and the image findings were compared on a segment-by-segment basis with the histopathologic findings. Sixty-four liver segments were analyzed, 20 of which contained tumor deposits. The overall sensitivity of PET, CT, and MRI was 30%, 25%, and 20%, respectively. For the detection of tumors with a histologic diameter exceeding 1 mm (n = 8), the sensitivity of PET, CT, and MRI was 63%, 38%, and 38%, respectively. The overall specificity of PET, CT, and MRI was 98%, 100%, and 93%, respectively. This study showed encouraging detectability and sensitivity for preclinical imaging of small liver tumors and provides valuable information on the imaging techniques for designing future protocols.


Editorial: Costing the N.H.S.


title: Structural and functional relationship between the receptor recognition and neuraminidase activities of the Newcastle disease virus hemagglutinin-neuraminidase protein: receptor recognition is dependent on neuraminidase activity.

text: The terminal globular domain of the paramyxovirus hemagglutinin-neuraminidase (HN) glycoprotein spike has a number of conserved residues that are predicted to form its neuraminidase (NA) active site, by analogy to the influenza virus neuraminidase protein. We have performed a site-directed mutational analysis of the role of these residues in the functional activity of the Newcastle disease virus (NDV) HN protein. Substitutions for several of these residues result in a protein lacking both detectable NA and receptor recognition activity. Contribution of NA activity, either exogenously or by coexpression with another HN protein, partially rescues the receptor recognition activity of these proteins, indicating that the receptor recognition deficiencies of the mutated HN proteins result from their lack of detectable NA activity. In addition to providing support for the homology-based predictions for the structure of HN, these findings argue that (i) the HN residues that mediate its NA activity are not critical to its attachment function and (ii) NA activity is required for the protein to mediate binding to receptors.


title: [Combating infectious disease using mathematical modelling].

text: When determining interventions against threatening infectious diseases such as HIV-infection, severe acute respiratory syndrome (SARS), smallpox and pandemic influenza, the use of mathematical models of the spread of infectious diseases is becoming increasingly popular. These models contribute to the structuring of the knowledge already available in various disciplines, to finding epidemiological connnections, to demonstrating lacunas within the pool of knowledge and to the comparison of the expected effects and costs of preventative and intervention measures. The use of models leads to a 'made-to-measure' analysis ofthe effects and costs of preventative and intervention measures which takes account of the specific characteristics of infectious diseases. The integration of knowledge from various disciplines can be supported by more research into the theoretical epidemiology of infectious disease and by better integration of mathematical models into policy development. The resulting and better foundations of this policy that are achieved by means of infectious disease modelling translate into more effective combating of infectious disease.


title: [A study of pulmonary inflammatory reaction induced by N-protein of SARS-CoV in rat models and effects of glucocorticoids on it].

text: OBJECTIVE To study the pulmonary inflammatory reaction induced by N-protein of SARS-CoV in rat models and the effects of glucocorticoids on the inflammatory reaction. METHODS The pulmonary inflammatory reaction in rat models were induced by intratracheal instillation of N-protein of SARS-CoV with a dose of 0.2 mg/kg. The rats were randomly divided into four groups: normal saline control group (Nc group), N-protein group 1 (P1 group, 6 h), N-protein group 2 (P2 group, 24 h), and N-protein + dexamethasone group (P + D group, dexamethasone 10 mg/kg intraperitoneally). The blood samples, bronchial alveolar lavage fluid (BALF) and lung tissue were collected after challenge. The cytological and histopathologic changes of lung tissues were observed and the wet/dry ratios (W/D) of lung tissue were determined. The interleukin (IL)-6, IL-10 and transforming growth factor-beta1 (TGF-beta1) of serum and BALF were measured by ELISA. RESULTS (1) Compared with the percentage of peripheral blood lymphocytes in Nc group [(68.42 +/- 13.07)%], that in P2 group [(50.50 +/- 14.36)%] was significantly decreased (P < 0.05); compared with Nc group and P2 group, that in P + D group was furthermore significantly decreased (P < 0.01). Compared with the total WBC of peripheral blood in Nc group [(5.86 +/- 2.25) x 10(9)] and P2 group [(4.83 +/- 1.49) x 10(9)], that in P + D group [(1.96 +/- 1.30) x 10(9)] was significantly decreased (P < 0.01). (2) Compared with the total WBC of BALF in Nc group [(95 +/- 29) x 10(7)], that in P2 group [(160 +/- 60) x 10(7)] was significantly increased (P < 0.05); but compared with P2 group, that in P + D group [(62 +/- 23) x 10(7)] was significantly decreased (P < 0.05). Analysis of BALF differential cell counts showed that the majority of cells were alveolar macrophages in all groups. (3) The W/D ratios of lung tissue in both P1 and P2 group [(5.18 +/- 0.29) and (5.19 +/- 0.34), respectively] after N-protein challenge were significantly increased than that in Nc groups [(4.77 +/- 0.27), P < 0.05]; the W/D ratio in P + D group (4.70 +/- 0.18) was significantly decreased than that in P2 group (P < 0.01). (4) Compared with Nc group, the levels of IL-6, IL-10, TGF-beta1 in both serum and BALF of P1 group were significantly increased (P < 0.01), and the levels of these cytokines in P2 group were significantly higher than those in P1 group (P < 0.01), but significantly lower in P + D group compared with P2 group (P < 0.01). CONCLUSIONS The N-protein of SARS-CoV had pathogenicity and could induce obvious pulmonary inflammatory reaction and acute lung injury, which were related to the increase and imbalance of pro-inflammatory and anti-inflammatory cytokines. Glucocorticoids could effectively alleviate the pulmonary inflammatory reaction induced by N-protein of SARS-CoV.


Transcription, replication, recombination, and engineering of coronavirus genes.


title: Infectivity of human coronavirus strain 229E.

text: The replication of human coronavirus strain 229E was observed by using indirect immunofluorescence in infected monolayers of MRC continuous cells. By 8 h after infection, bright cytoplasmic fluorescence was detected in cells infected with human coronavirus 229E. Discrete foci of infection were observed from 8 to 16 h after infection in cells infected with high dilutions of human coronavirus 229E; each fluorescent focus corresponded to a single virus infection. A fluorescent focus assay is described, using indirect immunofluorescence, which is more sensitive than the established techniques of tube titration and plaque assay. Particle/infectivity ratios for unpurified and purified virus preparations revealed a considerable drop in infectivity on purification.


Induction of apoptosis in MRC-5, diploid human fetal lung cells after infection with human coronavirus OC43.


title: Human rights?

text: The right to a fair trial is a fundamental freedom enshrined in the universal declaration of human rights. moreover there is a UN treaty, the covenant on civil and political rights, which contains detailed provisions concerning rights of fair trial. more than 140 nations have signed this treaty, making it the global yardstick for the protection of suspects.


title: Radioisotope cisternography: a potentially useful tool for headache diagnosis in patients with an IDD pump.

text: BACKGROUND Spontaneous intracranial hypotension (SIH) and related headache after intrathecal pump implantation is a possibility. Knowing the exact location of the cerebrospinal fluid (CSF) leak is mandatory for managing with interventional therapies. OBJECTIVES Radioisotope cisternography (RICG) is a common procedure in nuclear medicine and may be used as an additional procedure in the differential diagnosis of headaches due to intracranial liquor hypotension syndrome caused by microleakage of CSF. STUDY DESIGN We report an imaging case of complex chronic headache in a patient with an implanted drug delivery pump. The symptomatology of the patient was considered typical of spontaneous intracranial hypotension (SIH). Therefore, the initial suspicion was the loss of CSF due to the implanted pump catheter. The negativity of the routine headache test can exclude any leakage of CSF at the site of catheter insertion. SETTING Anesthesia, Critical Care, and Multidisciplinary Pain Management Department, General University General Hospital, Valencia, Spain. RESULTS Many patients are treated with implanted intrathecal infusion systems. In these patients, any neurological symptoms are readily attributed to the invasive neuroaxial system, especially severe cephalalgia. RICG was helpful in this particular case to make a diagnosis of exclusion of the diagnosis of SIH due to CSF leakage. CONCLUSION RICG may be considered as an additional diagnostic technique in patients with chronic pain and implanted spinal infusion devices for whom standard detection procedures have been exhausted.


title: Relationship between exhaled nitric oxide and airway hyperresponsiveness following experimental rhinovirus infection in asthmatic subjects.

text: Exhaled nitric oxide (NO) is elevated in asthmatics, and varies with disease severity. We postulated that a respiratory virus infection increases exhaled NO levels in asthma, and examined the relationship between the virus-induced changes in exhaled NO and in airway hyperresponsiveness to histamine. In a parallel study, seven patients underwent experimental rhinovirus 16 (RV16) inoculation at days 0 and 1, whilst seven patients received placebo. Exhaled NO was measured at baseline (day 0) and at days 1, 2 and 3 after inoculation. Histamine challenges were performed prior to (day -7) and after inoculation (day 3), and were expressed as provocative concentration causing a 20% fall in forced expiratory volume in one second (FEV1) (PC20). Following RV16 infection there was a significant increase in NO at days 2 and 3 as compared to baseline (median change (range): 4.2 (7.5) parts per billion (ppb), p=0.03, and 3.0 (10.1) ppb, p=0.02, respectively). Furthermore, PC20 decreased significantly following RV16 infection (mean+/-SD change in doubling dose: -0.65+/-0.54, p=0.02), whereas PC20 did not change in the placebo group (p=0.1). There was a significant correlation between the RV16-induced changes in exhaled NO levels at day 2 and the accompanying changes in PC20 at day 3 (rank correlation coefficient (rs): 0.86, p=0.01). Hence, the greater the increase in exhaled NO, the smaller the decrease in PC20. We conclude that rhinovirus infection increases exhaled nitric oxide levels in asthmatics, and that this increase is inversely associated with worsening of airway hyperresponsiveness to histamine. These results suggest that viral induction of nitric oxide synthase within the airways may play a protective role in exacerbations of asthma.


title: Safety and efficacy of laparoendoscopic single-site surgery for abdominal wall hernias.

text: BACKGROUND Laparoendoscopic single-site surgery has rapidly progressed from the animal laboratory to clinical use since mass production of multichannel ports began in 2007. Indeed, it has now been shown to be feasible and safe for many commonly performed operations. METHODS This study cohort comprised 22 unselected patients with abdominal wall hernias of varying types: multiply recurrent inguinal (n = 2), suprapubic (n = 1), ventral/incisional (n = 17), and parastomal hernias (n = 2), who underwent laparoendoscopic single-site ventral hernia repair between December 2009 and February 2011. Standard dissecting instruments and a 52 cm/5.5 mm/ 30 degree angle laparoscope were used. RESULTS Patients included 14 men and 8 women, with a median age of 56 (range, 32 to 78) years and a mean body mass index of 31.5 +/- 4.7 kg/m2. The mean mesh size was 460 cm2 (range, 225 to 884 cm2). Mean operation time was 125 minutes for ventral/incisional hernias and 270 minutes for parastomal hernias. No conversions to multiport or open surgeries were necessary. There was no mortality or morbidity, and no recurrence at 6- to 18-month follow-up. The mean satisfaction score was 2.7 (range, 2 to 3) with no patients reporting dissatisfaction with the procedure. CONCLUSION This series, though relatively small, represents a diverse group of patients with varying abdominal wall hernias, including parastomal hernias. These successful laparoendoscopic single-site surgeries, with no complications, demonstrate safety and efficacy, albeit in a specialized hernia center. This study is a prelude to the eventual validation of laparoendoscopic single-site hernia surgery with prospective randomized controlled trials.


title: Generation of coronavirus spike deletion variants by high-frequency recombination at regions of predicted RNA secondary structure.

text: Coronavirus RNA evolves in the central nervous systems (CNS) of mice during persistent infection. This evolution can be monitored by detection of a viral quasispecies of spike deletion variants (SDVs) (C. L. Rowe, S. C. Baker, M. J. Nathan, and J. O. Fleming, J. Virol. 71:2959-2969, 1997). We and others have found that the deletions cluster in the region from 1,200 to 1,800 nucleotides from the 5' end of the spike gene sequence, termed the "hypervariable" region. To address how SDVs might arise, we generated the predicted folding structures of the positive- and negative-strand senses of the entire 4,139-nt spike RNA sequence. We found that a prominent, isolated stem-loop structure is coincident with the hypervariable region in each structure. To determine if this predicted stem-loop is a "hot spot" for RNA recombination, we assessed whether this region of the spike is more frequently deleted than three other selected regions of the spike sequence in a population of viral sequences isolated from the CNS of acutely and persistently infected mice. Using differential colony hybridization of cloned spike reverse transcription-PCR products, we detected SDVs in which the hot spot was deleted but did not detect SDVs in which other regions of the spike sequence were exclusively deleted. Furthermore, sequence analysis and mapping of the crossover sites of 25 distinct patterns of SDVs showed that the majority of crossover sites clustered to two regions at the base of the isolated stem-loop, which we designated as high-frequency recombination sites 1 and 2. Interestingly, the majority of the left and right crossover sites of the SDVs were directly across from or proximal to one another, suggesting that these SDVs are likely generated by intramolecular recombination. Overall, our results are consistent with there being an important role for the spike RNA secondary structure as a contributing factor in the generation of SDVs during persistent infection.


No association of p53 codon 72 and p21 codon 31 polymorphisms in Taiwan Chinese patients with pterygium.


title: Antiviral drug discovery against SARS-CoV.

text: Severe Acute Respiratory Syndrome (SARS) is a life-threatening infectious disease caused by SARS-CoV. In the 2003 outbreak, it infected more than 8,000 people worldwide and claimed the lives of more than 900 victims. The high mortality rate resulted, at least in part, from the absence of definitive treatment protocols or therapeutic agents. Although the virus spreading has been contained, due preparedness and planning, including the successful development of antiviral drugs against SARS-CoV, is necessary for possible reappearance of SARS. In this review, we have discussed currently available strategies for antiviral drug discovery and how these technologies have been utilized to identify potential antiviral agents for the inhibition of SARS-CoV replication. Moreover, progress in the drug development based on different molecular targets is also summarized, including 1) Compounds that block the S protein-ACE2-mediated viral entry; 2) Compounds targeting SARS-CoV M(pro); 3) Compounds targeting papain-like protease 2 (PLP2); 4) Compounds targeting SARS-CoV RdRp; 5) Compounds targeting SARS-CoV helicase; 6) Active compounds with unspecified targets; and 7) Research on siRNA. This review aims to provide a comprehensive account of drug discovery on SARS. The experiences with the SARS outbreak and drug discovery would certainly be an important lesson for the drug development for any new viral outbreaks that may emerge in the future.


Characterisation of animal angiotensin-converting enzyme 2 receptors and use of pseudotyped virus to correlate receptor binding with susceptibility of SARS-CoV infection.


Patient-Doctor Telemedicine: Virtual Care in the Era of COVID-19 and Beyond.


Salmonella typhimurium brain abscess.


title: COVID-19 Pneumonia in Asymptomatic Trauma Patients; Report of 8 Cases.

text: We are currently involved in the novel coronavirus 2019 (COVID-19) pandemic. A considerable number of COVID-19 infected cases are asymptomatic but they can transmit the disease to others, especially healthcare workers. In this study, we reported 8 incidentally detected cases of COVID-19 pneumonia in chest computed tomography (CT) scan of patients referred to emergency department following multiple trauma without any respiratory symptoms.


title: Avian and pandemic influenza: an overview.

text: Influenza A/H5N1 (avian influenza) has now caused 258 human infections (as of November 13, 2006), with an approximate 50% mortality rate. Because the virus is novel in terms of antigenic type and causes infection and illness, and because humans have no pre-existing immunity, the conditions for a possible pandemic exist. Additionally, wild migratory birds appear to be spreading the virus across ever larger geographic areas, and newer clade 2 influenza A/H5N1 viruses have begun to emerge. The US Congressional Budget Office has formally modeled the likely consequences of pandemic influenza and estimates that up to 2 million of the US population might die, with up to 40% of all workers ill for as long as 3 or more weeks. This brief overview will review basic virologic, immunologic and epidemiologic information relevant to understanding and preparing for this threat. In particular, the role of avian influenza vaccines will be reviewed.


title: Recovery of 2'-deoxycoformycin-inhibited adenosine deaminase of mouse erythrocytes and leukemia L1210 in vivo.

text: The antibiotic 2'-deoxycoformycin, a potent inhibitor of adenosine deaminase, has potential as a chemotherapeutic agent. Injection of 2'-deoxycoformycin i.v. (0.2 mg/kg) to mice bearing ascites L1210 leukemia cells completely inhibits adenosine deaminase in both erythrocytes and L1210 cells. The recovery of the enzymic activity is markedly different in the two tissues. The recovery is very slow in erythrocytes (13% in 48 hr), whereas 80% recovery occurs during the same time interval in L1210 cells. This marked difference in the recovery of the enzyme in different tissues may play a role in the pharmacological and chemotherapeutic behavior of this drug.


Chinese authorities on alert as SARS breaks out again.


Hiatal Hernia Cruroplasty with a Running Barbed Suture Compared to Interrupted Suture Repair.


title: A 68-nucleotide sequence within the 3' noncoding region of simian hemorrhagic fever virus negative-strand RNA binds to four MA104 cell proteins.

text: The 3' noncoding region (NCR) of the negative-strand RNA [3'(-)NCR RNA] of the arterivirus simian hemorrhagic fever virus (SHFV) is 209 nucleotides (nt) in length. Since this 3' region, designated 3'(-)209, is the site of initiation of full-length positive-strand RNA and is the template for the synthesis of the 5' leader sequence, which is found on both full-length and subgenomic mRNAs, it is likely to contain cis-acting signals for RNA synthesis and to interact with cellular and viral proteins to form replication complexes. Gel mobility shift assays showed that cellular proteins in MA104 S100 cytoplasmic extracts formed two complexes with the SHFV 3'(-)209 RNA, and results from competition gel mobility shift assays demonstrated that these interactions were specific. Four proteins with molecular masses of 103, 86, 55, and 36 kDa were detected in UV-induced cross-linking assays, and three of these proteins (103, 55, and 36 kDa) were also detected by Northwestern blotting assays. Identical gel mobility shift and UV-induced cross-linking patterns were obtained with uninfected and SHFV-infected extracts, indicating that the four proteins detected are cellular, not viral, proteins. The binding sites for the four cellular proteins were mapped to the region between nt 117 and 184 (68-nt sequence) from the 3' end of the SHFV negative-strand RNA. This 68-nt sequence was predicted to form two stem-loops, SL4 and SL5. The 3'(-)NCR RNA of another arterivirus, lactate dehydrogenase-elevating virus C (LDV-C), competed with the SHFV 3'(-)209 RNA in competition gel mobility shift assays. UV-induced cross-linking assays showed that four MA104 cellular proteins with the same molecular masses as those that bind to the SHFV 3'(-)209 RNA also bind to the LDV-C 3'(-)NCR RNA and equine arteritis virus 3'(-)NCR RNA. However, each of these viral RNAs also bound to an additional MA104 protein. The binding sites for the MA104 cellular proteins were shown to be located in similar positions in the LDV-C 3'(-)NCR and SHFV 3'(-)209 RNAs. These data suggest that the binding sites for a set of the cellular proteins are conserved in all arterivirus RNAs and that these cell proteins may be utilized as components of viral replication complexes.


title: Canine distemper and related diseases: report of a severe outbreak in a kennel.

text: An outbreak of canine distemper in a kennel of German shepherds in the province of Bari is reported. Six 42-day-old pups developed typical signs of canine distemper (fever, conjunctivitis, respiratory distress and enteritis) and died within 7-10 days. Neurological symptoms were observed only in one pup. Four additional pups, which had shown no sign of illness, were separated and vaccinated, but two of these developed a severe, fatal nervous form 15 days later. Post-mortem examination, carried out on two pups which died without neurological signs, showed pneumonia and enteritis, more severe in one of the two examined pups. Smears from the brain and the conjunctiva of both dogs tested positive for canine distemper virus (CDV) by an immunofluorescent assay, confirmed by the identification of viral RNA using RT-PCR. Bordetella bronchiseptica and a canine adenovirus strain, characterized as canine adenovirus type 2 by a differential PCR assay, were isolated from the lungs of the pup showing the most pronounced lesions. Furthermore, canine coronavirus was detected by PCR in the intestinal content of this pup, suggesting a multifactorial aetiology of the outbreak.


title: Sixty-two severe and critical patients with 2009 influenza A (H1N1) in Shanghai, China.

text: BACKGROUND Pandemic influenza A (H1N1) emerged rapidly in China in May 2009. Preliminary comparisons with seasonal influenza suggest that pandemic 2009 influenza A (H1N1) disproportionately affects younger ages and causes generally mild disease. To characterize disease progress, comorbidities, and treatment outcomes among consecutive severe and critically ill patients in a hospital served as a reference center for the care of patients with H1N1 in Shanghai, China. METHODS A retrospective study on 62 severe and critically ill patients with 2009 influenza A (H1N1) was conducted in Shanghai Public Health Clinical Center. Demographic data, symptoms, comorbidities, disease progression, treatments, and clinical outcomes were collected for analysis. RESULTS Sixty-two severe or critically ill patients were admitted to the hospital with confirmed 2009 influenza A (H1N1) infection. The median age of the study cohort was 40 years old with a range from 18 years to 75 years, and 67.7% were males. All patients presented with fever and respiratory symptoms. At presentation, 34 patients (54.8%) had comorbidities such as smoking (29.0%), hypertension (29.0%) and hepatitis B virus infection (9.7%). The median time from symptom onset to hospital admission was 6 days (interquartile-range 3 - 14 days) and 23 critically ill patients were admitted to Intensive Care Unit after admission. All the patients received neuraminidase inhibitors (oseltaminir), while 60 patients (96.7%) were treated with antibiotics, and 39 (62.9%) with corticosteroids. Twenty-three critical cases received noninvasive mechanical ventilation on the first day of admission, and 3 of them ultimately required invasive ventilation. Four death reports (6.5%) were filed within the first 14 days from the onset of critical illness with the primary causes of severe acute respiratory distress syndrome, hypoxemia, or complications, secondary infection and sepsis, pyopneumothorax and stroke. CONCLUSIONS Severe illness from 2009 influenza A (H1N1) infection in Shanghai occurred among young individuals. Critical cases were associated with severe hypoxemia, multisystem organ failure, and a requirement for mechanical ventilation. Most patients had a good prognosis.


title: Diarylheptanoids from Alnus japonica inhibit papain-like protease of severe acute respiratory syndrome coronavirus.

text: The papain-like protease (PL(pro)), which controls replication of the severe acute respiratory syndrome coronavirus (SARS-CoV), has been identified as a potential drug target for the treatment of SARS. An intensive hunt for effective anti-SARS drugs has been undertaken by screening for natural product inhibitors that target SARS-CoV PL(pro). In this study, diarylheptanoids 1-9 were isolated from Alnus japonica, and the inhibitory activities of these compounds against PL(pro) were determined. Of the isolated diarylheptanoids, hirsutenone (2) showed the most potent PL(pro) inhibitory activity, with an inhibitory concentration (IC(50)) value of 4.1 µM. Structure-activity analysis showed that catechol and α,β-unsaturated carbonyl moiety in the molecule were the key requirement for SARS-CoV cysteine protease inhibition.


title: Occurrence and Management Strategies for Catheter Entrapment with Onyx Liquid Embolization.

text: In June 2012, Food and Drug Administration (FDA) issued a warning about the risk of catheter entrapment associated with Onyx embolization. We used our experience, literature review, and FDA Manufacturer and User Facility Device Experience (MAUDE) data review to identify five strategies to address catheter entrapment: 1/. Surgical resection of vessel at point of entrapment of catheter and retraction from exterior portion at the femoral region; 2/. Advancing and closing the loop of snare over the entrapped catheter followed by retraction; 3/. Advancing the distal access catheter over the entrapped catheter and retraction with forward movement of the distal access catheters; 4/. Inflation of balloon catheter coaxial to the entrapped catheter with subsequent retraction; and 5/. Intravascular retention and internalization of microcatheter. In the MAUDE data, there were 77 reports of catheter entrapment with Onyx embolization; microcatheter was retracted by surgical excision in 15, endovascular snare or other retriever devices in 5, deliberately entrapped inside the vessel using stent in 1, and left without intervention within intravascular compartment in 27 patients.


title: Evaluation of the efficiency of medical masks and the creation of new medical masks.

text: The effectiveness of medical masks in preventing respiratory infection was investigated by testing bacterial leakage, filtration efficiency, respiratory resistance and oxygen concentration of the enclosed space. Polypropylene (PP) fibres were treated with dimethyldioctadecylammonium bromide to impart a positive electrical charge capable of attracting bacteria. The fluffed PP fibres were used to make a polypropylene mask and to edge standard surgical and N-95 respirators to prevent leakage. A PP napkin was created by melting and blowing PP. The PP edging seal dramatically reduced bacterial leakage of standard masks and was more effective than adhesive paper tape edging in reducing respiratory resistance. Bacterial or viral filtration efficiency was almost 100% for the PP mask and the PP napkin. The specially designed PP mask with a synthetic adhesive at the edge of the mask may be more effective than the standard surgical mask and the N-95 respirator. The PP napkin is an important tool in preventing the spread of pathogens.


title: Use of an autogenous cortical graft in combination with guided tissue regeneration for treatment of an infrabony defect.

text: Infrabony periodontal defects are common findings encountered during complete oral examinations. Treatment options for infrabony lesions are aimed at meeting client demands as well as patient needs. Deciding on how to treat these lesions depends on the nature and degree of disease present as well as having the materials available to improve the chances of achieving the greatest clinical success. Bone grafting of an infrabony defect of the left mandibular first molar tooth of a dog using an autogenous cortical graft harvested with a reusable bone grafter in combination with guided tissue regeneration is described.


title: In vivo and in vitro characterization of an RNA replication enhancer in a satellite RNA associated with turnip crinkle virus.

text: RNA replication enhancers are cis-acting elements that can stimulate replication or transcription of RNA viruses. Turnip crinkle virus (TCV) and satC, a parasitic RNA associated with TCV infections, contain stem-loop structures that are RNA replication enhancers (P. Nagy, J. Pogany, and A. E. Simon, EMBO J. 1999, 18, 5653-5665). We have found that replacement of 28 nt of the satC enhancer, termed the motif1-hairpin, with 28 randomized bases reduced satC accumulation 8- to 13-fold in Arabidopsis thaliana protoplasts. Deletion of single-stranded flanking sequences at either side of the hairpin also affected RNA accumulation with combined alterations at both sides of the hairpin showing the most detrimental effect in protoplasts. In vitro analysis with a partially purified TCV RdRp preparation demonstrated that the motif1-hairpin in its minus-sense orientation was able to stimulate RNA synthesis from the satC hairpin promoter (located at the 3' end of plus strands) by almost twofold. This level of RNA synthesis stimulation is approximately fivefold lower than that observed with a linear promoter, suggesting that a highly stable hairpin promoter is less responsive to the presence of the motif1-hairpin enhancer than a linear promoter. The motif1-hairpin in its plus-sense orientation was only 60% as active in enhancing transcription from the hairpin promoter. Since the motif1-hairpin is a hotspot for RNA recombination during plus-strand synthesis and since satC promoters located on the minus-strand are all short linear sequences, these findings support the hypothesis that the motif1-hairpin is primarily involved in enhancing plus-strand synthesis.


title: [Enhanced-real time PCR: a highly sensitive method for SARS-coronavirus detection].

text: An enhanced real-time polymerase chain reaction (ERT-PCR) assay to detect the coronavirus associated with severe acute respiratory syndrome (SARS-Cov) has been designed for detection of SARS-Cov with high sensitivity and easy-to-interpret results, in which a target gene pre-amplification step preceded TaqMan real-time fluorescent PCR. The limit of detection of the ERT-PCR method was 10(-2) higher than the standard real-time PCR assay and 10(-7) higher than conventional PCR methods. The increased sensitivity of the assay would have major benefits in screening suspected SARS patients rapidly and efficiently and may help control the spread of SARS and other infectious diseases during future outbreak.


title: Restrictive visitor policies: feedback from healthcare workers, patients and families.

text: With the hospital-based transmission of Severe Acute Respiratory Syndrome (SARS) in Ontario, acute care hospitals severely restricted visitor access. Now that the SARS outbreak is under control, hospitals struggle with the balance between adhering to patient- and family-centred care models, and addressing the future threat of infectious diseases. To evaluate the effect of visitor restrictions and to guide future visitation policies, the Ottawa Hospital conducted a preliminary survey of patients, next of kin, staff, physicians and volunteers. Ninety percent of staff surveyed supported some form of visitor restrictions, while 71% indicated that they felt comfortable asking visitors to leave if they had exceeded current restrictions. The majority of patients (80%) and next of kin (76%) were at least moderately satisfied with current restricted limiting hours. A disproportionate number of positive comments on current visiting restrictions were received from both patients and staff. In the absence of evidence on which to base future visitor policy development, objective input from healthcare workers, patients and families is invaluable.


title: Multicenter quality assessment of PCR methods for detection of enteroviruses.

text: We conducted a multicenter evaluation of commercial and in-house PCR methods for the detection of enteroviruses. Three coded panels of test and control RNA samples, artificial clinical specimens, and representative enterovirus serotypes were used to assess amplification methods, RNA extraction methods, and reactivities with different enterovirus serotypes. Despite several differences between PCR methods, there was good agreement, although some variation in sensitivity was observed. Most PCR methods were able to detect enterovirus RNA derived from 0.01 50% tissue culture infective dose (TCID50) and were able to detect at least 1 TCID50 of enterovirus in cerebrospinal fluid, stool, or throat swab specimens. Most were also able to detect a wide range of enterovirus serotypes, although serotypic identification was not possible. Some laboratories experienced false-positive results due to PCR contamination, which appeared to result mainly from cross-contamination of specimens during RNA extraction. Provided that this problem is overcome, these PCR methods will prove to be a sensitive and rapid alternative to cell culture for the diagnosis of enterovirus infection.


title: [Electron microscopic detection rate of enteral viruses in diarrhea of dogs, cats, calves, swine and foals in the year 1988--electron microscopic study results].

text: During 1988 fecal and gut samples of 641 dogs, 198 cats, 576 calves, 108 piglets and 64 foals with diarrhoea were investigated for virus infections by electron microscopy. In samples of dogs and cats parvovirus was detected at a proportion of 21.9% and 16.7%, respectively; rotavirus alone or together with coronavirus was found only in 0.3-1.5% of the specimens. In samples of calves rotavirus, as well as coronavirus dominated with a detection rate amounting to 17.4% and 26.6% respectively (including 4.5% of mixed infections); parvovirus was present in a ratio of 0.5%. Specimens of piglets mainly contained coronavirus (25.0%), and in lower percentages rotavirus (2.8%), rota- and coronaviruses (0.9%) and parvovirus (0.9%). In feces of foals rotavirus was detected in 6.3% and particles resembling picornavirus in 4.7% of cases. Not identifiable virus particles resembling corona-or picornaviruses were rarely found (between 0.6-2.5) also in specimens of the other animal species.


title: Specific inhibition of coxsackievirus B3 translation and replication by phosphorothioate antisense oligodeoxynucleotides.

text: The 5' and 3' untranslated regions (UTRs) of coxsackievirus B3 (CVB3) RNA form highly ordered secondary structures that have been confirmed to play important regulatory roles in viral cap-independent internal translation initiation and RNA replication. We previously demonstrated that deletions in different regions of the 5' UTR significantly reduced viral RNA translation and infectivity. Such observations suggested strongly that viral RNA translation and replication could be blocked if highly specific antisense oligodeoxynucleotides (AS-ODNs) were applied to target crucial sites within the 5' and 3' UTRs. In this study, seven phosphorothioate AS-ODNs were synthesized, and the antiviral activity was evaluated by Lipofectin transfection of HeLa cells with AS-ODNs followed by infection of CVB3. Analysis by Western blotting, reverse transcription-PCR, and viral plaque assay demonstrated that viral protein synthesis, genome replication, and infectivity of CVB3 were strongly inhibited by the AS-ODNs complementary to different regions of the 5' and 3' UTRs. The most effective sites are located at the proximate terminus of the 5' UTR (AS-1), the proximate terminus of the 3' UTR (AS-7), the core sequence of the internal ribosome entry site (AS-2), and the translation initiation codon region (AS-4). These AS-ODNs showed highly sequence-specific and dose-dependent inhibitory effects on both viral protein synthesis and RNA replication. It is noteworthy that the highest inhibitory activities were obtained with AS-1 and AS-7 targeting the termini of the 5' and 3' UTRs. The percent inhibition values of AS-1 and AS-7 for CVB3 protein VP1 synthesis and RNA replication were 70.6 and 79.6 for AS-1 and 73.7 and 79.7 for AS-7, respectively. These data suggest that CVB3 infectivity can be inhibited effectively by AS-ODNs.


title: Isolation of Tettnang coronavirus from man?

text: A virus, identified as Tettnang virus, was isolated from the cerebrospinal fluid (CSF) of an 18 months old child with pharyngitis accompanied by an encephalitic reaction. The isolation of virus was followed by seroconversion. The aetiological role of the virus in the given disease is discussed.


title: Evaluation of a multiplexed PCR assay for detection of respiratory viral pathogens in a public health laboratory setting.

text: There are numerous viral and bacterial causes of respiratory disease. To enable rapid and sensitive detection of even the most prevalent causes, there is a need for more-simplified testing systems that enable researchers and clinicians to perform multiplexed molecular diagnostics quickly and easily. To this end, a new multiplexed molecular test called the MultiCode-PLx respiratory virus panel (PLx-RVP) was developed and then implemented in a public health laboratory setting. A total of 687 respiratory samples were analyzed for the presence of 17 viruses that commonly cause respiratory disease. As a comparator, the samples were also tested using a standard testing algorithm that included the use of a real-time influenza virus A and B reverse transcription-PCR test and routine viral culture identification. The standard testing algorithm identified 503 (73%) samples as positive and 184 as negative. Analyzing the same 687 samples, the PLx-RVP assay detected one or more targets in 528 (77%) samples and found 159 samples negative for all targets. There were 25 discordant results between the two systems; 14 samples were positive for viruses not routinely tested for by the Wisconsin State Laboratory of Hygiene, and 13 of these were confirmed by real-time PCR. When the results of the standard testing algorithm were considered "true positives," the PLx-RVP assay showed an overall sensitivity of 99% and an overall specificity of 87%. In total, the PLx-RVP assay detected an additional 40 viral infections, of which 11 were mixed infections.


title: Four major antigenic sites of the coronavirus transmissible gastroenteritis virus are located on the amino-terminal half of spike glycoprotein S.

text: Four major antigenic sites have been delineated on the spike protein (S) of the porcine enteric coronavirus transmissible gastroenteritis virus (TGEV) in previous topological studies using monoclonal antibodies (MAbs). Correlation of these sites with the physical structure of the protein was achieved by use of different approaches. Recombinant pEX plasmids directing the synthesis of various fused S polypeptides were constructed. A hybrid protein containing nine S-specific residues (363 to 371) was shown to express site C epitopes. The other sites were localized through study of the antigenic activity of fragments generated by controlled cleavage of the native protein with different endopeptidases. Two identified cleavage products of 26K and 13K, immunoreactive to site A-B- and site D-specific MAbs respectively, could be aligned on the S primary structure according to N-terminal sequence data. This led us to propose that the major neutralization domain A-B is contained in a region of approximately 200 residues with residue 506 as its N boundary. Similarly, site D epitopes should be located within a stretch of 130 residues, starting at 82 residues from the N terminus. Point mutations identified by direct RNA sequencing of neutralization-resistant mutants were consistent with the proposed location of these sites.


title: One-year epidemiology of fever at the Emergency Department.

text: BACKGROUND Although fever is recognised as a major presentation symptom at Emergency Departments (EDs) and is often used as a rationale for the institution of antibiotics, few studies describing patients with fever as the sole inclusion criterion at the ED of a general hospital have been performed. The objective of this study is to describe epidemiology of non-surgical febrile patients at the ED and to identify risk factors for adverse outcome. METHODS Blood, sputum, urine and faeces cultures, urine sediments and throat swaps for viral diagnostics were obtained from febrile ED patients. Outcome parameters were bacterial/viral infection, non-bacterial/non-viral infection, non-infectious febrile disease; mortality, hospital admission, admission to the intensive care unit (ICU) and length of hospital stay. RESULTS 213 Patients were included (87.8% were hospitalised, 8.5% were admitted to ICU, 4.2% died). In 75 patients (35.2%), bacterial infection was confirmed; in 78 patients (36.6%) bacterial infection was suspected. In nine patients (4.2%), viral diagnosis was confirmed; in six patients (2.8%), a viral condition was suspected. The most frequently encountered infection was bacterial pneumonia (58 patients, 27.2%). Only older age was correlated with mortality (ρ=0.176, p=0.01). CONCLUSION A majority of the febrile patients were admitted to the hospital, mostly for bacterial infection. An overall mortality rate of 4.2% was registered. Only a few risk factors for adverse outcome could be identified in this cohort. Overall, the outcome of patients presenting with fever at the ED is rather benign.


title: treatment and prevention of otitis media.

text: OBJECTIVE To review and summarize recent advances in the treatment and prevention of otitis media (OM). DATA SOURCES A MEDLINE search (1996-March 2000) was performed to identify relevant primary and review articles. References from these articles were also reviewed if deemed important. STUDY SELECTION AND DATA EXTRACTION English-language primary and review articles focusing on the treatment and prevention of acute otitis media (AOM) were included. Studies focusing exclusively on OM with effusion or serous OM and chronic suppurative OM were excluded. Information regarding prevention and drug therapy was reviewed, with an emphasis placed on advances made in the last two years. DATA SYNTHESIS Recently, an expert panel of the Centers for Disease Control and Prevention recommended use of only three of 16 systemic antibiotics approved by the Food and Drug Administration for treatment of AOM: amoxicillin, cefuroxime axetil, and ceftriaxone. Controversy exists over the importance of key selection factors used by the expert panel in determining which antibiotics to recommend in a two-step treatment algorithm, that is, in vitro data, pharmacodynamic profiles, and necessity for coverage of drug-resistant Streptococcus pneumoniae at all steps of empiric treatment. Additional antibiotic and patient selection factors useful for individualizing therapy include clinical efficacy, adverse effects, frequency and duration of administration, taste, cost, comorbid infections, and ramifications should bacterial resistance develop to the chosen antibiotic. Presumed or past patient/caregiver adherence (especially when antibiotic failure has occurred) is also paramount in selecting antibiotic therapy. A three-step treatment algorithm for refractory AOM that employs amoxicillin, trimethoprim/sulfamethoxazole (TMP/SMX), or high-dose amoxicillin/clavulanate (depending on the prior dose of and adherence to amoxicillin therapy), and ceftriaxone or tympanocentesis at steps 1, 2, and 3, respectively, appears rational and cost-effective. The recent upsurge in antimicrobial resistance is highlighted, and recommendations are presented for the treatment of AOM and prevention of recurrent otitis media (rAOM). CONCLUSIONS Amoxicillin remains the antibiotic of choice for initial empiric treatment of AOM, although the traditional dosage should be increased in patients at risk for drug-resistant S. pneumoniae. In cases refractory to high-dose amoxicillin, TMP/SMX should be prescribed if adherence to prior therapy seemed good or complete, or high-dose amoxicillin/clavulanate if adherence was incomplete or questionable. Ceftriaxone should be reserved as third-line treatment. The increasing prevalence of drug-resistant S. pneumoniae emphasizes the importance of alternative medical approaches for the prevention of OM, as well as judicious antibiotic use in established cases. Removal of modifiable risk factors should be first-line therapy for prevention of rAOM. We support the use of conjugate pneumococcal vaccine per guidelines for prevention of rAOM from the Advisory Committee on Immunization Practice of the Centers for Disease Control and Prevention, with consideration given to influenza vaccine for cases of rAOM that historically worsen during the flu season. Sulfisoxazole prophylaxis should be reserved for children who are immunocompromised, have concurrent disease states exacerbated by AOM, or meet the criteria of rAOM despite conjugate pneumococcal and influenza vaccination. Therapy should be intermittent, beginning at the first sign of an upper respiratory infection, and should continue for 10 days. The invasive nature and risks of anesthesia relegate myringotomy, tympanostomy tubes, and adenoidectomy to last-line therapies for rAOM.


Pediatric Dentistry During and After COVID-19.


title: Innovation and challenges in funding rapid research responses to emerging infectious diseases: Lessons learned from the outbreak of severe acute respiratory syndrome.

text: Although the local public health response to the severe acute respiratory syndrome outbreak in Canada was critical to the diagnosis, management and treatment of patients, such a rapid research response required a national effort to engage the research and stakeholder communities. The Canadian research effort, coordinated through the Institute of Infection and Immunity of the Canadian Institutes of Health Research and the Michael Smith Foundation for Health Research, has provided insight into the mechanisms required to ensure the rapid development of strategical initiatives in response to emerging infectious diseases. It has also provided a rational basis to set up a national network to be engaged if needed in the future.


The new International Health Regulations: an historic development for international law and public health.


New WHO chief pledges to expand network to stem disease outbreaks.


title: Fgl2 deficiency causes neonatal death and cardiac dysfunction during embryonic and postnatal development in mice.

text: We hypothesized that cardiac dysfunction was responsible for the high perinatal lethality that we previously reported in fibrinogen-like protein 2 (Fgl2) knockout (KO) mice. We therefore used ultrasound biomicroscopy to assess left ventricular (LV) cardiac structure and function during development in Fgl2 KO and wild-type (WT) mice. The only deaths observed between embryonic day (E)8.5 (onset of heart beating) and postnatal day (P)28 (weaning) were within 3 days after birth, when 33% of Fgl2 KO pups died. Histopathology and Doppler assessments suggested that death was due to acute congestive cardiac failure without evidence of valvular or other obvious cardiac structural abnormalities. Heart rates in Fgl2 KO embryos were significantly reduced at E8.5 and E17.5, and irregular heart rhythms were significantly more common in Fgl2 KO (21/26) than WT (2/21) embryos at E13.5. Indexes of systolic and/or diastolic cardiac function were also abnormal in KO mice at E13.5 and E17.5, in postnatal mice studied at P1, and in KO mice surviving to P28. M-mode analysis showed no difference in LV diastolic chamber dimension, although posterior wall thickness was thinner at P7 and P28 in Fgl2 KO mice. We conclude that Fgl2 deficiency is not associated with obvious structural cardiac defects but is associated with a high incidence of neonatal death as well as contractile dysfunction and rhythm abnormalities during embryonic and postnatal development in mice.


title: Structure and intracellular assembly of the transmissible gastroenteritis coronavirus.

text: Coronaviruses have been described as pleomorphic, round particles with a helical nucleocapsid as the unique internal structure under the virion envelope. Our studies on the organization of the transmissible gastroenteritis coronavirus (TGEV) have shown that the structure of these viruses is more complex. Different electron microscopy techniques, including cryomicroscopy of vitrified viruses, revealed the existence of an internal core, most probably icosahedral, in TGEV virions. Disruption of these cores induced the release of elongated ribonucleoprotein complexes. Ultrastructural analysis of freeze-substituted TGEV-infected swine testis (ST) cells showed characteristic intracellular budding profiles as well as two types of virions. While large virions with an electron-dense internal periphery are seen at perinuclear regions, smaller viral particles exhibiting compact internal cores of poligonal contours are more abundant in areas closer to the plasma membrane of the cell. These data strongly suggest that maturation events following the budding process are responsible for the formation of the internal core shell, the new structural element that we have recently described in extracellular infectious TGEV virions.


title: Severe acute respiratory syndrome: overview with an emphasis on the Toronto experience.

text: OBJECTIVE To provide an overview of the severe acute respiratory syndrome (SARS) outbreak in Toronto, Ontario, which experienced the largest outbreak outside Asia, and to review what has been learned during the past year. DATA SOURCES MEDLINE search of all studies related to SARS, including review of the Centers for Disease Control and Prevention, World Health Organization (WHO), and Health Canada Web sites. DATA SYNTHESIS During the SARS outbreak in Toronto, 438 people had been diagnosed as having suspected or probable SARS and 44 people died. Elderly people and those with comorbid illnesses were at greatest risk of complications or death. Transmission was via direct contact with respiratory secretions. The use of gloves, gowns, N95 masks, and eye protection was effective in preventing transmission. No transmission occurred before symptom onset or after recovery. Serologic tests suggest that antibodies may not appear until 28 days after illness onset. Molecular tests give their greatest yield during the second week of illness. The value of ribavirin treatment remains questionable. The combination of interferon plus corticosteroids appears to be better than corticosteroids alone. Postmortem examination revealed pulmonary edema and evidence of diffuse alveolar damage. Very few morphological changes were noted in other organs despite the presence of viral RNA as detected by polymerase chain reaction. CONCLUSION On July 5, 2003, the WHO declared that the SARS outbreak was over. Since then, new cases of SARS have been reported in Asia. With global travel, the disease can rapidly spread throughout the world. Therefore, we must remain vigilant to prevent another pandemic.


title: COVID-19: Striking a Balance between Health Care Provider Safety and Patient Care in the Indian Hospital Settings.

text: Background The COVID-19 pandemic has taken its toll on the health care systems all over the world. The global shortage of resources has adversely affected the safety of healthcare personnel as well patients. This has forced the health care facilities to adopt various measures and improvisation in infection control practices, in addition to those in personnel protective equipments (PPEs) adapting to their need and their resources. Objectives To review the literature on the infection control practices in health care facilities against COVID-19 and put forward practical solutions, in addition to personnel protective equipments that may be adopted for health care personnel and patient safety in the Indian setting. Conclusions We are now amidst of COVID-19 pandemic and we need to focus on 'Risk reduction as much as possible'. The facilities and resources should transform to meet the challenges within the shortest time frame. The concept of patient care needs to balance with the safety of the healthcare, patient and society. This is achievable only by a combination of engineering controls, administrative controls and PPEs. It involves a lot of committed team work while managing an outbreak like COVID - 19 attached with a lot of social stigma.


title: Structures of proteases for ubiqutin and ubiquitin-like modifiers.

text: Post-translational modifiers can alter the function of proteins in many different ways. The conjugation of ubiquitin (Ub) and ubiqutin-like modifiers (Ubls) to proteins has been shown to be especially crucial in regulating a variety of cellular processes including the cell cycle, growth control, quality control, localization and many more. It is a highly dynamic process and involves a number of enzymes called E1, E2 and E3. Ub and Ubls are removed from the target proteins by deubiquitinating enzymes (DUBs) or Ubl-specific proteases (ULPs), thereby deconjugation can act as an additional level of control over the ubiquitin-conjugation system. In addition, DUBs and ULPs are responsible for activating Ub and Ubls from their inactive corresponding precursor forms. Here we review recent progress in molecular details of these deconjugating enzymes of Ubls.


title: Organizational culture influences health care workers' influenza immunization behavior.

text: BACKGROUND AND OBJECTIVES Low rates of influenza immunization among health care workers (HCWs) pose a potential health risk to patients in primary care practices. Despite previous educational efforts and programs to reduce financial barriers, HCW influenza immunization rates remain low. Variation in practice-level organizational culture may affect immunization rates. To explore this relationship, we examined organizational cultures and HCWs' influenza immunization behaviors in three family medicine practices. METHODS We used a multi-method comparative case study. A field researcher used participant observation, in-depth interviews, and key informant interviews to collect data in each practice in November-December 2003. A diverse team used grounded theory to analyze text data. RESULTS Organizational culture varied among practices and differing HCW immunization rates were observed. The most structured and business-like practice achieved immunization of all HCWs, while the other two practices exhibited greater variation in HCW immunization rates. Physicians in the practices characterized as chaotic/disorganized or divided were immunized at higher rates than other members of the practices. CONCLUSIONS In these practices, organizational culture was associated with varying rates of influenza immunization for HCWs, especially among nonphysicians. Addressing elements of organizational culture such as beliefs regarding influenza immunization and office policies may facilitate the immunization of all staff members.


title: The ethics of responding to a novel pandemic.

text: Recent epidemics and pandemics have highlighted a number of ethical concerns about the response to the increasing threat of emerging infectious diseases. Some of these ethical concerns are very fundamental. They include why a pandemic was declared, how much clinical information can be collected for public health without threatening patient confidentiality and how to ensure fairness in the distribution of resources. We discuss these issues and suggest approaches to resolve these dilemmas as we anticipate the next pandemic.


title: Comparison of targeting F and G protein genes to detect bovine and ovine respiratory syncytial viruses.

text: In this study, 2 reverse transcriptase-polymerase chain reaction (RT-PCR) assays were developed and compared for simultaneous detection of bovine and ovine respiratory syncytial viruses (RSVs). One assay was based on a set of primers, which amplified a 426-bp fragment of either bovine or ovine RSV F gene (RT-PCR F). The F products could be distinguished by EcoRI or BstYI restriction endonuclease cleavage. In the other assay, a set of primers amplified a 542-bp fragment of either ovine or bovine RSV G gene (RT-PCR G). EcoO1091 and RsaI restriction enzymes were used to differentiate between the ovine and bovine PCR-G products. Sequencing of the PCR products confirmed the fidelity of both assays. The 2 assays were evaluated using 18 bovine RSV isolates, 1 ovine RSV, 1 bighorn sheep RSV isolate, 1 caprine RSV isolate, 2 human RSV isolates, and several other viruses associated with bovine respiratory tract disease. RT-PCR G may be more sensitive in detecting viral RNA. Because the target sequence of the F gene is more conserved than that of the G gene, RT-PCR F followed by the appropriate restriction enzyme cleavage may be superior to RT-PCR G to discriminate between the 2 ruminant RSV subgroups. This assay should prove useful for determining the relative contribution of ovine and bovine RSV to the pathogenesis of bovine respiratory tract disease.


title: Coronavirus and Pasteurella infections in bovine shipping fever pneumonia and Evans' criteria for causation.

text: Respiratory tract infections with viruses and Pasteurella spp. were determined sequentially among 26 cattle that died during two severe epizootics of shipping fever pneumonia. Nasal swab and serum samples were collected prior to onset of the epizootics, during disease progression, and after death, when necropsies were performed and lung samples were collected. Eighteen normal control cattle also were sampled at the beginning of the epizootics as well as at weekly intervals for 4 weeks. Respiratory bovine coronaviruses (RBCV) were isolated from nasal secretions of 21 and 25 cattle before and after transport. Two and 17 cattle nasally shed Pasteurella spp. before and after transport, respectively. RBCV were isolated at titers of 1 x 10(3) to 1.2 x 10(7) PFU per g of lung tissue from 18 cattle that died within 7 days of the epizootics, but not from the lungs of the remaining cattle that died on days 9 to 36. Twenty-five of the 26 lung samples were positive for Pasteurella spp., and their CFU ranged between 4.0 x 10(5) and 2.3 x 10(9) per g. Acute and subacute exudative, necrotizing lobar pneumonia characterized the lung lesions of these cattle with a majority of pneumonic lung lobes exhibiting fibronecrotic and exudative changes typical of pneumonic pasteurellosis, but other lung lobules had histological changes consisting of bronchiolitis and alveolitis typical of virus-induced changes. These cattle were immunologically naive to both infectious agents at the onset of the epizootics, but those that died after day 7 had rising antibody titers against RBCV and Pasteurella haemolytica. In contrast, the 18 clinically normal and RBCV isolation-negative cattle had high hemagglutinin inhibition antibody titers to RBCV from the beginning, while their antibody responses to P. haemolytica antigens were delayed. Evans' criteria for causation were applied to our findings because of the multifactorial nature of shipping fever pneumonia. This analysis identified RBCV as the primary inciting cause in these two epizootics. These viruses were previously not recognized as a causative agent in this complex respiratory tract disease of cattle.


title: Coronaviruses as vectors: position dependence of foreign gene expression.

text: Coronaviruses are the enveloped, positive-stranded RNA viruses with the largest RNA genomes known. Several features make these viruses attractive as vaccine and therapeutic vectors: (i) deletion of their nonessential genes is strongly attenuating; (ii) the genetic space thus created allows insertion of foreign information; and (iii) their tropism can be modified by manipulation of the viral spike. We studied here their ability to serve as expression vectors by inserting two different foreign genes and evaluating systematically the genomic position dependence of their expression, using a murine coronavirus as a model. Renilla and firefly luciferase expression cassettes, each provided with viral transcription regulatory sequences (TRSs), were inserted at several genomic positions, both independently in different viruses and combined within one viral genome. Recombinant viruses were generated by using a convenient method based on targeted recombination and host cell switching. In all cases high expression levels of the foreign genes were observed without severe effects on viral replication in vitro. The expression of the inserted gene appeared to be dependent on its genomic position, as well as on the identity of the gene. Expression levels increased when the luciferase gene was inserted closer to the 3' end of the genome. The foreign gene insertions generally reduced the expression of upstream viral genes. The results are consistent with coronavirus transcription models in which the transcription from upstream TRSs is attenuated by downstream TRSs. Altogether, our observations clearly demonstrate the potential of coronaviruses as (multivalent) expression vectors.


title: Renin-angiotensin system, hypertension, and chronic kidney disease: pharmacogenetic implications.

text: About 80% of CKD (chronic kidney disease) patients are hypertensive, and kidney function and blood pressure are clearly related to both physiologic and pathologic conditions in a "vicious cycle". In this pathologic scenario, there is a renin-angiotensin system (RAS) hyperactivity associated to progression of renal damage. Current guidelines indicate as the first choice of antihypertensive intervention, the pharmacologic blockade of the RAS. Nonetheless, both response to treatment and renal protection have considerable inter-individual variability. The main aims of this review are to describe the genetic characteristics of RAS components and to identify the possible pharmacogenetic implications for RAS-blocker drugs in the hypertension-CKD scenario. To date, RAS polymorphisms have not been consistently associated to antihypertensive response and studies focusing on CKD are scarce. Nonetheless, pharmacogenetic studies for the RAS-blocker drugs could still be further explored, especially with new generation tools and focusing not only on the antihypertensive response, but also on renal protection as well.


Severe acute respiratory syndrome (SARS)--150 days on.


title: Laparoscopic and mini-invasive surgery.

text: BACKGROUND JGCA Gastric Cancer Treatment Guidelines (2004) include Laparoscopic Assisted Distal Gastrectomy (LADG) within the chapter of modified surgery. A metanalysis published in 2010 shows that LADG is significantly superior to Open Distal Gastrectomy (ODG) if comparing short term outcomes. Oncologic results prove to be comparable to ODG by one RCT and 2 retrospective studies. Little evidence is available on Laparoscopic Total Gastrectomy and concerns are raised about long-term oncologic outcomes. MATERIALS AND METHODS Laparoscopic Subtotal Gastrectomy is carried out with 4 to 5 ports in the periumbilical region (Hasson trocar for laparoscope) and upper quadrants. After exploration of the abdominal cavity surgical steps include coloepiploic detachment, omentectomy, dissection of the gastrocolic ligament, division of the left gastroepiploic vessels, division of right gastroepiploic vessels, division of pyloric vessels. The duodenum is transected with a linear stapler. Incision of the lesser omentum and dissection of the hepatoduodenal ligament allows completion of D2 lymphadenectomy. The 4/5ths of the stomach are transected starting from the greater curve at the junction of left and right gastroepiploic arcades by linear stapler. Roux-en-Y loop reconstruction is performed through a stapled side-to-side gastro-jejunal anastomosis and a side-to-side jejuno-jejunal anastomosis. Reconstruction after Laparoscopic Total Gastrectomy is performed preferably by a side-to-side esophago-jejunal anastomosis according to Orringer. DISCUSSION AND CONCLUSION A robotic assisted approach adds precision on lymphadenectomy and reconstructive techniques.


title: Assuring public health professionals are prepared for the future: the UAB public health integrated core curriculum.

text: In response to calls to improve public health education and our own desire to provide a more relevant educational experience to our Master of Public Health students, the University of Alabama at Birmingham (UAB) School of Public Health designed, developed, and instituted a fully integrated public health core curriculum in the fall of 2001. This curriculum combines content from discipline-specific courses in biostatistics, environmental health, epidemiology, health administration, and the social and behavioral sciences, and delivers it in a 15 credit hour, team-taught course designed in modules covering such topics as tobacco, infectious diseases, and emergency preparedness. Weekly skills-building sessions increase student competence in data analysis and interpretation, communication, ethical decision-making, community-based interventions, and policy and program planning. Evaluations affirm that the integrated core is functioning as intended: as a means to provide critical content in the core disciplines in their applied context. As public health education continues to be debated, the UAB public health integrated core curriculum can serve as one model for providing quality instruction that is highly relevant to professional practice.


title: Regulation of transcription of coronaviruses.

text: To study factors involved in regulation of transcription of coronaviruses, we constructed defective interfering (DI) RNAs containing sg RNA promoters at multiple positions. Analysis of the amounts of sg DI RNA produced by these DIs resulted in the following observations: (i) a downstream promoter downregulates an upstream promoter; (ii) an upstream promoter has little or no effect on the activity of a downstream promoter. Our data suggest that attenuation of upstream promoter activities by downstream promoter sequences plays an important role in regulating the amounts of sg RNAs produced by coronaviruses. Our observations are in accordance with the models proposed by Konings et al. and Sawicki and Sawicki.


title: Severe acute respiratory syndrome: management and reconfiguration of a radiology department in an infectious disease situation.

text: Severe acute respiratory syndrome, or SARS, is a new infectious disease pandemic with important public health concerns. The high infectivity rate by means of droplet transmission places health care workers at substantial risk of contracting the disease. Radiology departments are particularly affected, since imaging plays a vital role in both diagnosis and follow-up of this disease. The authors outline their experience in infection control and isolation procedures during this outbreak. Barrier precautions, reconfiguration of the department, separation of imaging equipment, cleaning procedures, personal protective equipment, and staff safety are discussed.


Use of defective RNAs containing reporter genes to investigate targeted recombination for avian infectious bronchitis virus.


title: Evidence for emergence of an amphibian iridoviral disease because of human-enhanced spread.

text: Our understanding of origins and spread of emerging infectious diseases has increased dramatically because of recent applications of phylogenetic theory. Iridoviruses are emerging pathogens that cause global amphibian epizootics, including tiger salamander (Ambystoma tigrinum) die-offs throughout western North America. To explain phylogeographical relationships and potential causes for emergence of western North American salamander iridovirus strains, we sequenced major capsid protein and DNA methyltransferase genes, as well as two noncoding regions from 18 geographically widespread isolates. Phylogenetic analyses of sequence data from the capsid protein gene showed shallow genetic divergence (< 1%) among salamander iridovirus strains and monophyly relative to available fish, reptile, and other amphibian iridovirus strains from the genus Ranavirus, suggesting a single introduction and radiation. Analysis of capsid protein sequences also provided support for a closer relationship of tiger salamander virus strains to those isolated from sport fish (e.g. rainbow trout) than other amphibian isolates. Despite monophyly based on capsid protein sequences, there was low genetic divergence among all strains (< 1.1%) based on a supergene analysis of the capsid protein and the two noncoding regions. These analyses also showed polyphyly of strains from Arizona and Colorado, suggesting recent spread. Nested clade analyses indicated both range expansion and long-distance colonization in clades containing virus strains isolated from bait salamanders and the Indiana University axolotl (Ambystoma mexicanum) colony. Human enhancement of viral movement is a mechanism consistent with these results. These findings suggest North American salamander ranaviruses cause emerging disease, as evidenced by apparent recent spread over a broad geographical area.


title: A contribution for the forensic odontologist's safety in the autopsy room.

text: The Covid 19 pandemic we are currently facing all over the world is undoubtly an emergency condition which exposes all workers to the risks of a serious infection. The personnel working in the autoptic room is at high risk for the possible contact with infected material from corpses, splashes and aerosols diffusion, and must be always prepared to stay safe and healthy. We offer to the forensic odontologists' community a short summary of some old and new hints on the treatment of dead bodies in the autopsy room during this pandemic emergency time.