title: Avian flu from an occupational health perspective.

text: Although a pandemic scenario necessarily involves a situation in which a large number of people become infected in a nonoccupational setting, past infectious disease events have highlighted the unique role that occupational exposure may play in the emergence of disease in humans. While small-scale poultry farmers have already demonstrated their high risk of exposure and subsequent infection with H5N1 avian flu, other bird handling occupations may also be at high risk. In this review article, the author highlights the occupational health aspects of a newly emerging infectious disease, H5N1 avian flu, and the efforts being made by various authorities to confront the possibility of the next flu pandemic. Various measures potentially available to protect high-risk workers are outlined, emphasizing a hierarchy of controls in which these measures can be placed. The author postulates that, by reducing the risk of transmission of avian flu to these workers, occupational medicine can play a pivotal role in the world's effort to prepare for a pandemic, and may be uniquely positioned to contribute to the effort to delay or even avert the emergence of the next pandemic.


title: Implementation of an automated, real-time public health surveillance system linking emergency departments and health units: rationale and methodology.

text: In September 2004, Kingston, Frontenac, Lennox and Addington (KFL and A) Public Health, in collaboration with the Public Health Division of the Ontario Ministry of Health and Long-Term Care, Queen's University, the Public Health Agency of Canada, Kingston General Hospital and Hotel Dieu Hospital, began a 2-year pilot project to implement and evaluate an emergency department (ED) chief complaint syndromic surveillance system. Our objective was to evaluate a comprehensive and readily deployable real-time regional syndromic surveillance program and to determine its ability to detect gastrointestinal or respiratory outbreaks well in advance of traditional reporting systems. In order to implement the system, modifications were made to the University of Pittsburgh's Real-time Outbreak and Disease Surveillance (RODS) system, which has been successfully integrated into public health systems, and has enhanced communication and collaboration between them and EDs. This paper provides an overview of a RODS-based syndromic surveillance system as adapted for use at a public health unit in Kingston, Ontario. We summarize the technical specifications, privacy and security considerations, data capture, classification and management of the data streams, alerting and public health response. We hope that the modifications described here, including the addition of unique data streams, will provide a benchmark for future Canadian syndromic surveillance systems.


title: [Bioinformatics-based Design of Peptide Vaccine Candidates Targeting Spike Protein of MERS-CoV and Immunity analysis in Mice].

text: Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as a novel human coronavirus and posed great threat to public health world wide,which calls for the development of effective and safe vaccine urgently. In the study, peptide epitopes tagrgeting spike antigen were predicted based on bioinformatics methods. Nine polypeptides with high scores were synthesized and linked to keyhole limpet hemocyanin (KLH). Female BALB/C mice were immunized with individual polypeptide-KLH, and the total IgG was detected by ELISA as well as the cellular mediated immunity (CMI) was analyzed using ELIs-pot assay. The results showed that an individual peptide of YVDVGPDSVKSACIEVDIQQTFFDKTWPRPIDVSKADGI could induce the highest level of total IgG as well as CMI (high frequency of IFN-γ secretion) against MERS-CoV antigen in mice. Our study identified a promising peptide vaccine candidate against MERS-CoV and provided an experimental support for bioinformatics-based design of peptide vaccine.


title: Serotype identification of avian infectious bronchitis virus by RT-PCR of the peplomer (S-1) gene.

text: The S-1 peplomer gene sequences of 31 strains of avian coronavirus infectious bronchitis virus (IBV) from North America, Europe, and Australia were compared to identify common and unique regions for possible diagnostic applications. S-1 sequences that were conserved among serotypes and sequences that were variable between serotypes were identified. Based on conserved S-1 gene sequences, "general" degenerate oligonucleotide primers were designed that amplified IBV genomic RNA by the reverse transcriptase polymerase chain reaction (RT-PCR) procedure regardless of serotype. Primers specific for IBV serotypes Massachusetts, Connecticut, Arkansas, JMK, Delaware (DE/072/92), and California (CA/633/85) were designed from regions of the S-1 gene exhibiting extensive sequence hypervariability. The ability to identify these six serotypes of IBV by RT-PCR was demonstrated by testing the serotype-specific primers on a panel of unknown samples that included 30 reference strains and field isolates previously characterized by virus neutralization (VN). The use of serotype-specific primers in RT-PCR provides a rapid and accurate means of identifying IBV.


title: Protection of mammalian cells from severe acute respiratory syndrome coronavirus infection by equine neutralizing antibody.

text: The aetiological agent for severe acute respiratory syndrome (SARS) has been determined to be a new type of coronavirus (SARS-CoV) that infects a wide range of mammalian hosts. Up to now, there have been no specific drugs to protect against SARS-CoV infection, thus developing effective strategies against this newly emerged viral infection warrants urgent efforts. Adoptive immune therapy with pathogen-specific heterologous immunoglobulin has been successfully used to control the dissemination of many viral infections. To investigate whether a neutralizing antibody against SARS-CoV raised in an artiodactylous host can have a protective role on primate cells, we prepared serum IgGs and their pepsin-digested F(ab')2 fragments from horses inoculated with purified SARS-CoV (BJ-01 strain). The protective effect of the F(ab')2 fragments against SARS-CoV infection was determined in cultured Vero E6 cells by cytopathic effect (CPE), MTT and plaque-forming assays and in a Balb/c mouse model by CPE and quantitative RT-PCR. The results showed the neutralization titres of F(ab')2 from three horses all reached at least 1:1600, and 50 microg of the F(ab')2 fragments could completely neutralize 1x10(4) TCID50- SARS-CoV in vivo. Additionally, we observed that F(ab')2, against BJ-01 strain could also protect cells from infection by the variant GZ-01 strain in vitro and in vivo. Our work has provided experimental support for testing the protective equine immunoglobulin in future large primate or human trials.


title: Hematology and serum biochemistry of Sumatran rhinoceroses (Dicerorhinus sumatrensis) in a rainforest sanctuary in Way Kambas National Park, Indonesia.

text: There is a paucity of basic biological information for the Sumatran rhinoceros (Dicerorhinus sumatrensis). This information is fundamental to husbandry and management practices for captive animals and for support of in situ conservation efforts. Serial blood samples were collected over an 8-yr period to evaluate patterns in hematology and serum biochemistry values among five Sumatran rhinoceroses housed at the Sumatran Rhino Sanctuary in Way Kambas National Park, Lampung, Indonesia. Understanding the basis for variance in analytes both within and between subjects can allow use of more sensitive subject-based reference values, and is particularly suitable for small populations of endangered animals. Both intra- and intersubject variability was computed for each analyte and the associated index of individuality was determined. Previously published cutoff points for index of individuality indicate where population-based reference intervals can be used with confidence (index > 1.4) or with caution (0.6 < index < 1.4). Interrhino variability was small for the majority of analytes, with 12 of 19 analytes having an index of individuality greater than 1.4 and none having an index of individuality less than 0.6. With the high within-individual variability of most anayltes in the Sumatran rhinoceroses at the sanctuary, subject-based reference intervals offer little advantage over standard population-based reference intervals for monitoring the health of these endangered animals. Differences were noted (but not tested for statistical significance) in serum urea, aspartate aminotransferase (AST), lactate dehydrogenase (LDH), and cholesterol between young and old rhinoceroses, and in hematocrit, AST, alanine aminotransferase (ALT), LDH, and glucose between male and female rhinoceroses. Husbandry practices, animal management, nutrition and habitat factors may also impact hematology and biochemistry results, and these relationships deserve more careful investigation. This study represents the most comprehensive hematology and serum biochemistry comparison of Sumatran rhinoceroses held in natural rainforest conditions outside a traditional zoological setting.


title: Leader sequences of coronavirus are altered during infection.

text: The life cycle of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) involves a unique process called discontinuous transcription by which a set of 3' coterminal subgenomic mRNAs (sgmRNA) with identical 5' leader sequences can be generated. The current study demonstrates that the replication intermediates of minus strand of subgenomic RNA (sgRNA) can be readily recovered from SARS-CoV infected cells. A novel sgmRNA (M-1) was identified as a short version of membrane (M) gene. Transcriptional regulatory sequences (TRS) of SARS-CoV and Mouse Hepatitis Virus (MHV) sgmRNAs contain a species specific core element (CE). The sizes of leader sequences in MHVs vary not only in different viral strains but also among different genes in the same strain. Leader alterations such as deletion and nucleotide substitution were observed in MHVs, while a dynamic one-orientation 'sequential deletion' was found among the leaders of 76 SARS-CoV isolates. These results imply that the leader sequence of coronavirus might be unstable and leader alterations during SARS-CoV transmission in humans might have negative impact on its viral infectivity.


Porcine reproductive and respiratory syndrome.


title: Variation in the spike protein of the 793/B type of infectious bronchitis virus, in the field and during alternate passage in chickens and embryonated eggs.

text: The degree of variation exhibited within the 793/B serotype (also known as 4/91 and CR88 serotypes) was investigated with nine French and 10 British isolates, collected between 1985 and 1994. The S1 part (1644 nucleotides) of the spike protein gene of the first known isolate of this serotype, FR/CR85131/85, had 95.9% to 97% nucleotide identity with the other isolates. Partial sequencing of isolates from Iran and Saudi Arabia, isolated in 2000, revealed approximately 95% nucleotide identity with European isolates, including the two live 793/B vaccinal strains, showing that they were not re-isolations of vaccinal virus. The data indicates that strains within the 793/B serotype have > or =96% nucleotide identity within the whole S1 gene and > or =93% nucleotide identity within the first 560 nucleotides, and > or =92% and > or =86% amino acid identities in the corresponding protein regions. This is similar to the identities exhibited within the Massachusetts serotype. Sequence analysis of a 793/B field isolate after passage in embryonated eggs, then in chickens and then again in eggs revealed selection for a serine and alanine at S1 amino acid position 95 in chicken-passaged and egg-passaged virus, respectively. There was no change in pathogenicity. This is the first demonstration at gene sequence level of host-driven selection for infectious bronchitis virus.


title: Science and technology foresight: a provocative tool for contending with future challenges in food safety and public veterinary medicine.

text: The author describes how foresight methods can provide a provocative approach to some of the key challenges faced by the world of animal health and public veterinary practices while supporting the safety and security of the food system and public health in general. Being provocative is important because the future may be very unfamiliar and demands an approach of critical thinking, which can best be activated by having to consider the prospective reality of one or more substantially different policy operational environments. The factors that are shaping our future may also be quite disruptive (for example from 1989 to 1994 when the Cold War abruptly ended, the Soviet Union dissolved and the internet was born). Consequently it is essential that any forward preparedness efforts explore a range of plausible options and not immediately discount those that may in today's terms appear unlikely. The author reviews the methods through to the point where scenario parameters are defined, and then switches to observations about how the process can influence and provoke policy formulation. The results of the foresight and scenarios employment are described by Willis in another paper in this volume.


title: Virus-like particles in cystic mammary adenoma of a snow leopard.

text: Virus-like particles were observed in the giant cells of a mammary adenoma of a snow leopard kept in captivity. Particles that measured 115 to 125 nm in diameter budded from the lamella of endoplasmic reticulum and were studded on their inner surfaces with dense granules (approximately 12 nm) that gave them their unique ultrastructural morphology. Such particles were not observed extracellularly. Type B or type C particles were not seen in the tumor tissue.


title: Cefuroxime axetil versus clarithromycin in the treatment of acute maxillary sinusitis.

text: Acute maxillary sinusitis is a common condition requiring broad-spectrum therapy to prevent development of chronic disease. A randomised, double-blind, multicentre study was performed to compare the efficacy and safety of cefuroxime axetil 250 mg twice daily (n = 185) and clarithromycin 250 mg twice daily (n = 185), both administered for 10 days, in the treatment of patients with acute sinusitis. Efficacy was determined by assessment of clinical response at post-treatment and follow-up, and by radiological assessment at pre-treatment and follow-up. Assessment of days absent from work due to illness was also made. In the cefuroxime axetil group, 169/185 (91%) patients were cured/improved at post-treatment, as were 172/185 (93%) patients receiving clarithromycin and, of these, 137/169 (81%) and 143/172 (83%) maintained their response at follow-up. Follow-up radiography showed a reduction in incidence of air fluid level and/or opacification from 96% to 15% (cefuroxime axetil) and from 96% to 11% (clarithromycin), and a decrease in frequency of mucosal thickening from 58% to 28% (cefuroxime axetil) and from 56% to 29% (clarithromycin). Only 10% of patients in either group experienced adverse events and days absent from work were comparable. This study demonstrated clinical equivalence between twice-daily cefuroxime axetil and clarithromycin, both treatments being effective and well tolerated.


title: Improving surge capacity for biothreats: experience from Taiwan.

text: This article discusses Taiwan's experience in managing surge needs based on recent events, including the 1999 earthquake, severe acute respiratory syndrome in 2003, airliner crashes in 1998 and 2001, and yearly typhoons and floods. Management techniques are compared and contrasted with U.S. approaches. The authors discuss Taiwan's practices of sending doctors to the scene of an event and immediately recalling off-duty hospital personnel, managing volunteers, designating specialty hospitals, and use of incident management systems. The key differences in bioevents, including the mathematical myths regarding individual versus population care, division of stockpiles, the Maginot line, and multi-jurisdictional responses, are highlighted. Several recent initiatives aimed at mitigating biothreats have begun in Taiwan, but their efficacy has not yet been tested. These include the integration of the emergency medical services and health-facility medical systems with other response systems; the use of the hospital emergency incident command system; crisis risk-communications approaches; and the use of practical, hands-on training programs. Other countries may gain valuable insights for mitigating and managing biothreats by studying Taiwan's experiences in augmenting surge capacity.


Feline infectious peritonitis: a coronavirus disease of cats.


title: The autocatalytic release of a putative RNA virus transcription factor from its polyprotein precursor involves two paralogous papain-like proteases that cleave the same peptide bond.

text: The largest replicative protein of coronaviruses is known as p195 in the avian infectious bronchitis virus (IBV) and p210 (p240) in the mouse hepatitis virus. It is autocatalytically released from the precursors pp1a and pp1ab by one zinc finger-containing papain-like protease (PLpro) in IBV and by two paralogous PLpros, PL1pro and PL2pro, in mouse hepatitis virus. The PLpro-containing proteins have been recently implicated in the control of coronavirus subgenomic mRNA synthesis (transcription). By using comparative sequence analysis, we now show that the respective proteins of all sequenced coronaviruses are flanked by two conserved PLpro cleavage sites and share a complex (multi)domain organization with PL1pro being inactivated in IBV. Based upon these predictions, the processing of the human coronavirus 229E p195/p210 N terminus was studied in detail. First, an 87-kDa protein (p87), which is derived from a pp1a/pp1ab region immediately upstream of p195/p210, was identified in human coronavirus 229E-infected cells. Second, in vitro synthesized proteins representing different parts of pp1a were autocatalytically processed at the predicted site. Surprisingly, both PL1pro and PL2pro cleaved between p87 and p195/p210. The PL1pro-mediated cleavage was slow and significantly suppressed by a non-proteolytic activity of PL2pro. In contrast, PL2pro, whose proteolytic activity and specificity were established in this study, cleaved the same site efficiently in the presence of the upstream domains. Third, a correlation was observed between the overlapping substrate specificities and the parallel evolution of PL1pro and PL2pro. Collectively, our results imply that the p195/p210 autoprocessing mechanisms may be conserved among coronaviruses to an extent not appreciated previously, with PL2pro playing a major role. A large subset of coronaviruses may employ two proteases to cleave the same site(s) and thus regulate the expression of the viral genome in a unique way.


title: Attaching and effacing Escherichia coli infections in calves, pigs, lambs, and dogs.

text: Attaching and effacing Escherichia coli (AEEC) adhere to mucosal epithelium in both small and large intestine and induce a distinctive lesion characterized by an irregular scalloped appearance of the epithelial layer. Infection with attaching and effacing E. coli was detected in 14 calves, 7 pigs, 2 lambs, and 3 dogs. Affected animals were from farms and kennels in South Dakota, Minnesota, Iowa, Nebraska, and Wisconsin. Ages of affected animals were calves, 2 days to 4 months; pigs, 1-6 weeks; lambs, 1 week; and dogs, 7-8 weeks. Clinical signs included diarrhea in all animals, but other nonenteric disease problems were present in some animals. Concurrent infection with other enteropathogens was detected in 9 calves and 5 pigs. Infection with AEEC appeared to be the sole cause of illness and death in some animals. There was evidence of intestinal hemorrhage in 5 of the calves and in all 3 dogs. Attaching and effacing lesions varied from small scattered foci to widespread involvement of large areas of intestinal mucosa. Verotoxin was produced by E. coli strains isolated from 9 calves, but not by strains from pigs, lambs, or dogs.


title: Single-incision laparoscopic cholecystectomy: from four wounds to one.

text: OBJECTIVE To review the initial results and surgical outcomes of single-incision laparoscopic cholecystectomy. DESIGN Prospective case series. SETTING A university teaching hospital and a regional hospital in Hong Kong. PATIENTS All patients undergoing single-incision laparoscopic cholecystectomy from August 2009 to March 2011. RESULTS Fifty patients underwent single-incision laparoscopic cholecystectomy during the study period. The indications for surgery included symptomatic gallstones (n=43) and gallbladder polyps (n=7). The mean operating time was 78 (standard deviation, 24) minutes. Forty-five of the patients successfully underwent single-incision laparoscopic cholecystectomy, giving a success rate of 90%. In the remaining five patients, additional working ports were constructed to obtain better exposure and dissection around Calot's triangle. On comparing the results of the initial 25 cases to the subsequent 25 cases, in the latter group the operating time was significantly shorter (86 vs 71 minutes; P=0.02), and the success rate was higher (80% vs 100%; P=0.05). During the median follow-up period of 6.8 months, four patients had complications, which included: postoperative urinary retention (n=2), one each with a haematoma and an incisional hernia. No patient endured bile duct injury, postoperative bile leakage, or haemorrhage in our series. CONCLUSION Single-incision laparoscopic cholecystectomy is feasible and safe for treatment of uncomplicated gallbladder diseases. There was a reduction in the operating time and increase in success rate with accumulation of experience. Nevertheless, surgeons should be cautious about the potential risks of this new technique.


title: The Global Role of the World Health Organization.

text: The 21st century global health landscape requires effective global action in the face of globalization of trade, travel, information, human rights, ideas, and disease. The new global health era is more plural, comprising a number of key actors, and requiring more coordination of effort, priorities and investments. The World Health Organization (WHO) plays an essential role in the global governance of health and disease; due to its core global functions of establishing, monitoring and enforcing international norms and standards, and coordinating multiple actors toward common goals. Global health governance requires WHO leadership and effective implementation of WHO's core global functions to ensure better effectiveness of all health actors, but achieving this global mission could be hampered by narrowing activities and budget reallocations from core global functions.


Recent exposure to smoking and COVID-19.


title: Terror Australis 2004: preparedness of Australian hospitals for disasters and incidents involving chemical, biological and radiological agents.

text: OBJECTIVE To assess the level of preparedness of Australian hospitals, as perceived by senior emergency department physicians, for chemical, biological and radiological (CBR) incidents, as well as the resources and training available to their departments. METHODS Detailed questionnaires were mailed to the directors of the 86 hospital emergency departments (EDs) in Australia accredited by the Australasian College for Emergency Medicine. Questions covered hospital planning, available resources and training, and perceived preparedness. RESULTS Responses were received from 76 departments (88%): 73 reported that their ED had a disaster plan, with 60 (79%) having a contingency plan for chemical, 57 (75%) for biological, and 53 (70%) for radiological incidents. Specific staff training for managing patients from a conventional mass casualty incident was given in 83% of EDs, falling to 66% for a CBR incident. Forty-three per cent reported that their plan involved staff managing contaminated patients, but availability of personal protective equipment and decontamination facilities varied widely. Although 41% believed their ED could cope with a maximum of 20 patients in the first 2 hours after a conventional incident, this increased to 71% for a CBR incident. Staff training was considered the main funding priority (59%). CONCLUSIONS This survey raises significant questions about the level of preparedness of Australian EDs for dealing with patients from both conventional and CBR incidents. Hospitals need to review their plans and functionality openly and objectively to ensure that their perceived preparedness is consistent with reality. In addition, they urgently require guidance as to reasonable expectations of their capacity. To that end, we recommend further development of national standards in hospital disaster planning and preparedness.


title: Identification and characterization of dominant helper T-cell epitopes in the nucleocapsid protein of severe acute respiratory syndrome coronavirus.

text: By using a series of overlapping synthetic peptides covering 98% of the amino acid sequence of the nucleocapsid protein (NP) of severe acute respiratory syndrome coronavirus (SARS-CoV), four helper T-cell (Th) epitopes (NP11, residues 11 to 25; NP51, residues 51 to 65; NP61, residues 61 to 75; and NP111, residues 111 to 125) in C57BL mice (H-2(b)), four (NP21, residues 21 to 35; NP91, residues 91 to 105; NP331, residues 331 to 345; and NP351, residues 351 to 365) in C3H mice (H-2(k)), and two (NP81, residues 81 to 95; and NP351, residues 351 to 365) in BALB/c mice (H-2(d)) have been identified. All of these peptides were able to stimulate the proliferation of NP-specific T-cell lines or freshly isolated lymph node cells from mice immunized with recombinant NP. Immunization of mice with synthetic peptides containing appropriate Th epitopes elicited strong cellular immunity in vivo, as evidenced by delayed-type hypersensitivity. Priming with the helper peptides (e.g., NP111 and NP351) significantly accelerated the immune response induced by recombinant NP, as determined by the production of NP-specific antibodies. When fused with a conserved neutralizing epitope (SP1143-1157) from the spike protein of SARS-CoV, NP111 and NP351 assisted in the production of high-titer neutralizing antibodies in vivo. These data provide useful insights regarding immunity against SARS-CoV and have the potential to help guide the design of peptide-based vaccines.


title: Biosecurity measures to prevent the incursion of invasive alien species into Japan and to mitigate their impact.

text: The 2004 Japanese 'Invasive Alien Species Act' was enacted to control invasive alien species and prevent the damage that they cause to ecosystems. The Act defines invasive alien species as those recognised as, or suspected of, causing damage to ecosystems, human safety, agriculture, forestry and fisheries. Invasive alien species are carefully regulated: raising, planting, keeping or transporting them is prohibited without the express permission of the relevant minister. The Act represents a revolutionary advance for biological conservation in Japan. However, enforcing the Act is problematic. Dealing with the European bumblebee (Bombus terrestris), for example, involved resolving a bitter dilemma between biological conservation and agricultural productivity. The Act also has a serious loophole; it does not include alien micro-organisms. The incursion of amphibian chytridiomycosis into Japan caused confusion for scientists and the Japanese Government because control of such an alien micro-organism was not anticipated in the Act. Japan faces particular difficulties in attempting to control alien species because of its reliance on imports.


title: Fatal foodborne Clostridium perfringens illness at a state psychiatric hospital--Louisiana, 2010.

text: Clostridium perfringens, the third most common cause of foodborne illness in the United States (1), most often causes a self-limited, diarrheal disease lasting 12-24 hours. Fatalities are very rare, occurring in <0.03% of cases (1). Death usually is caused by dehydration and occurs among the very young, the very old, and persons debilitated by illness (2). On May 7, 2010, 42 residents and 12 staff members at a Louisiana state psychiatric hospital experienced vomiting, abdominal cramps, and diarrhea. Within 24 hours, three patients had died. The three fatalities occurred among patients aged 41-61 years who were receiving medications that had anti-intestinal motility side effects. For two of three decedents, the cause of death found on postmortem examination was necrotizing colitis. Investigation by the Louisiana Office of Public Health (OPH) and CDC found that eating chicken served at dinner on May 6 was associated with illness. The chicken was cooked approximately 24 hours before serving and not cooled in accordance with hospital guidelines. C. perfringens enterotoxin (CPE) was detected in 20 of 23 stool specimens from ill residents and staff members. Genetic testing of C. perfringens toxins isolated from chicken and stool specimens was carried out to determine which of the two strains responsible for C. perfringens foodborne illness was present. The specimens tested negative for the beta-toxin gene, excluding C. perfringens type C as the etiologic agent and implicating C. perfringens type A. This outbreak underscores the need for strict food preparation guidelines at psychiatric inpatient facilities and the potential risk for adverse outcomes among any patients with impaired intestinal motility caused by medications, disease, and extremes of age when exposed to C. perfringens enterotoxin.


title: Workforce integration of new graduate nurses: evaluation of a health human resources employment policy.

text: Historically, economic changes have negatively affected the nursing workforce in Ontario. The trend towards part-time and casual employment emerged from healthcare restructuring in the 1990s. The severe acute respiratory syndrome (SARS) outbreak in 2003 alerted the Ontario government to the issue of part-time and casual nursing. In 2007, the Nursing Graduate Guarantee (NGG), a health human resources employment policy, was developed as a financial incentive for employers to hire and mentor new graduate nurses for a six-month period. The purpose of this study was to examine facilitators and barriers to policy implementation and assess the impact of the NGG on full-time employment and workforce integration of new graduate nurses in Ontario. A mixed-methods approach was used and included surveys, interviews and focus groups. Results indicated that full-time employment of new graduate nurses increased during the study period and that mentorship facilitated workforce integration of new graduate nurses.


Primary Structure of the S peplomer gene of bovine coronavirus and surface expression in insect cells.


Male intergenerational intimacy: historical, socio-psychological, and legal perspectives.


title: [Prokaryotic expression of S2 extracellular domain of SARS coronavirus spike protein and its fusion with Hela cell membrane].

text: OBJECTIVE To construct the expression plasmid of S2 extracellular domain (S2ED) of SARS-coronavirus (SARS- Cov) spike protein (S protein) and enhanced green fluorescent protein (EGFP) to obtain the fusion protein expressed in prokaryotic cells. METHODS S2ED based on bioinformatics prediction and EGFP sequence were amplified by PCR and inserted into pET-14b plasmid. The recombinant protein His-S2ED-EGFP was expressed in E. coli by IPTG induction. After purification by Ni-NTA agarose beads, the soluble fractions of the fusion protein were collected and identified by SDS-PAGE and Western blotting. The fusion of S2ED with Hela cell membranes was observed with fluorescent microscope. RESULTS The pET-14b-S2ED-EGFP plasmid was correctly constructed and highly expressed in BL21 (DE3). When incubated with Hela cells, the purified protein could not internalize through membrane fusion. CONCLUSIONS The expression plasmid containing S2ED of SARS-Cov S protein and EGFP sequence is constructed successfully. Although the recombinant protein obtained has not shown the expected fusion effect with Hela cell membrane, this work may enrich the understanding of the process of membrane fusion mediated by S2 protein and lay the foundation for future study of targeting cell transport system based on cell-specific binding peptide.


Balamuthia amebic encephalitis risk, Hispanic Americans.


title: Sequence changes of infectious bronchitis virus isolates in the 3' 7.3 kb of the genome after attenuating passage in embryonated eggs.

text: Attenuated infectious bronchitis virus (IBV) vaccines are available but the relationship between sequence and virulence is not clear. In this study, the sequences of the 3' 7.3 kb of the genome, amplified using reverse transcription-polymerase chain reaction before and after attenuation, were compared to study the relationship between virulence and the sequences of three IBV strains. After attenuation, two to six amino acid substitutions were found in the spike 1 subunit, and two or three amino acid substitutions were found in the spike 2 subunit. None or one amino acid substitution was found in the small membrane protein, and one or three amino acid substitutions were found in the membrane protein. However, no amino acid substitution was found in the nucleocapsid (N) protein, indicating that the N protein might not be related to this attenuation. The 3' untranslated region after the N gene of one strain was partially deleted after attenuation, and might be correlated with virulence. This study is the first demonstration for IBV comparing sequence changes in the 3' 7.3 kb genome after attenuation. The aforementioned information on amino acid changes might be useful in future virulence studies.


title: [COVID-19 in the Emergency Department of Bernhoven hospital].

text: OBJECTIVE To describe disease presentation and clinical characteristics of patients with COVID-19 presenting to the Emergency Department (ED) of Bernhoven hospital in Uden, the Netherlands. DESIGN Prospective, descriptive study. METHOD The registry focused on atypical symptoms and co-infections. We hypothesized that patients older than 70 years more often have atypical symptoms. The number of co-infections is unknown. Therefore, we prospectively registered medical history, duration of symptoms, symptoms, temperature, lab results and co-infections of patients with confirmed COVID-19 in the period March 4-16th. RESULTS The clinical characteristics of 107 patients were registered. The average age was 71 years and 41% was female. The median duration of symptoms was 5 days. 19% of patients had not been referred to pulmonary or internal medicine. Symptoms were fever (78%), respiratory complaints (78%), chest pain (28%), abdominal pain (13%), and diarrhea (34%). In 54% of the COVID-19 patients at the ED, the temperature was ≥ 38,0°C, CRP ≥ 50 in 51%, leucocytosis in 12% and elevated LD in 61%. Of 31 patients 24 (77%) had an absolute lymphopenia. Co-infections were seen in 16% of patients. The mortality in the ED was 2% and ICU-admission 5%. On March 25th, 2020 the overall mortality was 22% and ICU-admission 15%. CONCLUSION We have seen patients with a very serious disease resulting in a high mortality and ICU-admission. Over 35% of patient did not have the typical symptoms of fever and respiratory complaints; atypical symptoms like chest pain, abdominal pain and diarrhea are frequently seen. There is no difference between patients over and under 70 years. COVID-19 patients can present with atypical symptoms, co-infections and distributed over various medical specialties.


title: Morning and evening plasma melatonin and dexamethasone suppression test in patients with nonseasonal major depressive disorder from northern Greece (latitude 40-41.5 degrees ).

text: INTRODUCTION The present study aimed to search for correlations between melatonin (MT) levels and the dexamethasone suppression test (DST) and clinical variables. METHODS Fifty depressed patients aged 21-60 years took part in the study. The Schedules for Clinical Assessment in Neuropsychiatry, version 2.0, and the International Personality Disorders Examination were used for diagnosis. Psychometric assessment included the Hamilton Depression Rating Scale, the Hamilton Anxiety Scale, the General Assessment of Funtioning Scale, the Newcastle scales and the Diagnostic Melancholia Scale. The DST and 9.00 and 23.00 h MT values were assessed. Statistical analysis included Student's t test, Pearson product moment correlation coefficient and forward stepwise multiple linear regression analysis. RESULTS Melancholic patients had lower 23.00 h MT values in comparison to the rest of the patients and the atypical and 'undifferentiated' patients. CONCLUSION The current study shows that low MT values were closely related to melancholic depression. Distinct quality of mood, psychomotor agitation or retardation and anorexia or weight loss seemed to be responsible for this relationship.


title: Causative pathogens of severe diarrhea in children.

text: OBJECTIVES To investigate the enteropathogens in children with diarrhea attending Salmaniya Medical Complex, Bahrain. METHODS Fecal samples from 805 children up to 15 years were examined for parasites, ova and cysts by direct wet preparation, formol-ether concentration and modified Ziehl-Neelsen stain, during the period November 1998 through to June 2000. Samples were cultured for Salmonella, Shigella, Campylobacter and Enteropathogenic Escherichia coli. Antibiotic sensitivity tests were performed on the relevant clinical isolates by agar disk diffusion method. All stools from children below 3 years of age (653 samples) were processed for adenovirus and rotavirus using a commercially available latex agglutination test (Diarlex). In addition, reverse transcription-polymerase chain reaction was performed on 200 randomly selected samples using oligonucleotide primers for Rotavirus A, B and C. RESULTS Four subjects were found positive for parasites. Eighty-three (10.3%) samples were found positive for Salmonella (46 isolates), Shigella (26 isolates), Campylobacter jejuni (7 isolates), and Enteropathogenic Escherichia coli (4 isolates). Rotavirus was found in 91 (13.9%) samples and 4 samples (0.6%) were found positive for adenovirus. Out of 200 samples examined by reverse transcription-polymerase chain reaction, 73 (36.5%) were positive for group A rotavirus. CONCLUSIONS Rotavirus type A appeared to be the most common single agent in our pediatric population, followed by the classical bacterial pathogens. Adenovirus and parasites appeared to play a very minor role in diarrhea. Thus, we suggest the introduction of rotavirus diagnostic tests in microbiological examination of diarrheic stools of children below 3 years of age.


title: SARS related preventive and risk behaviours practised by Hong Kong-mainland China cross border travellers during the outbreak of the SARS epidemic in Hong Kong.

text: OBJECTIVES To investigate patterns of behaviours and attitudes related to SARS prevention in the Hong Kong cross border traveller population. SETTINGS A survey was carried out at the Hong Kong-China cross border checkpoint in the middle of the epidemic. PARTICIPANTS A total of 839 Hong Kong adult residents returning to Hong Kong from mainland China were surveyed. MAIN OUTCOME MEASURES Practice of preventive measures and relevant behaviours and attitudes. RESULTS Around 40% of the respondents were using masks all or most of the time in public places or washing their hands frequently (>10 times per day) and about one third avoided visiting crowded places in mainland China. Such figures were however lower than those practised by the general public in Hong Kong. SARS related perceptions, such as perceived risk of transmission and efficacy, etc, were associated with mask use and not visiting crowded places, but not with hand washing, which was associated with duration of stay. Gender differences were also observed. Around 70% of the travellers would have delayed medical consultation for influenza-like illness in China; 12.7% would not wear masks during such episodes of illness. Furthermore, about 30% of the respondents used to wear masks in Hong Kong but not in mainland China. CONCLUSIONS The findings have implications on cross border prevention of SARS. It seems that those travelling during the SARS epidemic were a "self selected" group, and they were using less preventive measures. Special attention and intervention need to be provided to travellers to prevent a second wave cross border transmission of the disease.


title: Characterization of a monoclonal antibody and its single-chain antibody fragment recognizing the nucleoside Triphosphatase/Helicase domain of the hepatitis C virus nonstructural 3 protein.

text: We have produced a murine monoclonal antibody (MAb), ZX10, recognizing the NTPase/helicase domain of the hepatitis C virus (HCV) nonstructural 3 protein (NS3), from which we designed a single-chain variable fragment (ScFv). The ZX10 MAb recognized a discontinuous epitope of the NTPase/helicase domain, of which the linear sequence GEIPFYGKAIPL at residues 1371 to 1382 constitutes one part. cDNAs from variable regions coding for the heavy and light chains were cloned, sequenced, and assembled into the NS3-ScFv, which was inserted into procaryotic and eucaryotic expression vectors. Escherichia coli-expressed NS3-ScFv inhibited the binding of the ZX10 MAb to NS3, confirming a retained specificity. However, the ability to bind the peptide 1371-1382 had been lost. In vitro-translated NS3-ScFv and HCV NS3/NS4A were coprecipitated by antibodies to HCV NS4A, confirming the in vitro activity of the NS3 ScFv. Thus, we have designed a functional NS3 NTPase/helicase domain-specific ScFv which should be evaluated further with respect to disturbing enzymatic functions of the NS3 protein.


title: Emerging infections and old friends: remaining prepared in South Dakota.

text: Recent reports of serious infection outbreaks internationally remind us of the importance of accurate information and continual vigilance. The Ebola outbreak in West Africa has captured headlines as the most severe outbreak in the history of this disease. West Nile disease, measles, pertussis and tuberculosis infect South Dakota patients on a yearly basis. A significant rise in syphilis cases has prompted recommendations for increased prenatal screening. The more unusual viral diseases, Ebola, Middle East respiratory syndrome (MERS) and Chikungunha virus, receive media attention but present minimal risk to the state, while the annual influenza epidemic continues to plague us all. We review these infections, both old and emerging, and describe national and local preparedness practices.


Vigorous controls minimise risk.


title: Effect of intergenic consensus sequence flanking sequences on coronavirus transcription.

text: Insertion of a region, including the 18-nucleotide-long intergenic sequence between genes 6 and 7 of mouse hepatitis virus (MHV) genomic RNA, into an MHV defective interfering (DI) RNA leads to transcription of subgenomic DI RNA in helper virus-infected cells (S. Makino, M. Joo, and J. K. Makino, J. Virol. 66:6031-6041, 1991). In this study, the subgenomic DI RNA system was used to determine how sequences flanking the intergenic region affect MHV RNA transcription and to identify the minimum intergenic sequence required for MHV transcription. DI cDNAs containing the intergenic region between genes 6 and 7, but with different lengths of upstream or downstream flanking sequences, were constructed. All DI cDNAs had an 18-nucleotide-long intergenic region that was identical to the 3' region of the genomic leader sequence, which contains two UCUAA repeat sequences. These constructs included 0 to 1,440 nucleotides of upstream flanking sequence and 0 to 1,671 nucleotides of downstream flanking sequence. An analysis of intracellular genomic DI RNA and subgenomic DI RNA species revealed that there were no significant differences in the ratios of subgenomic to genomic DI RNA for any of the DI RNA constructs. DI cDNAs which lacked the intergenic region flanking sequences and contained a series of deletions within the 18-nucleotide-long intergenic sequence were constructed to determine the minimum sequence necessary for subgenomic DI RNA transcription. Small amounts of subgenomic DI RNA were synthesized from genomic DI RNAs with the intergenic consensus sequences UCUAAAC and GCUAAAC, whereas no subgenomic DI RNA transcription was observed from DI RNAs containing UCUAAAG and GCTAAAG sequences. These analyses demonstrated that the sequences flanking the intergenic sequence between genes 6 and 7 did not play a role in subgenomic DI RNA transcription regulation and that the UCUAAAC consensus sequence was sufficient for subgenomic DI RNA transcription.


title: Scrub typhus and tropical rickettsioses.

text: PURPOSE OF REVIEW Recent developments in molecular taxonomic methods have led to a reclassification of rickettsial diseases. The agent responsible for scrub typhus (Orientia tsutsugamushi ) has been removed from the genus Rickettsia and a bewildering array of new rickettsial pathogens have been described. An update of recent research findings is therefore particularly timely for the nonspecialist physician. RECENT FINDINGS An estimated one billion people are at risk for scrub typhus and an estimated one million cases occur annually. The disease appears to be re-emerging in Japan, with seasonal transmission. O. tsutsugamushi has evolved a variety of mechanisms to remain viable in its intracellular habitat. Slowing the release of intracellular calcium inhibits apoptosis of macrophages. Subsets of chemokine genes are induced in infected cells, some in response to transcription factor activator protein 1. Cardiac involvement is uncommon and clinical complications are predominantly pulmonary. Serious pneumonitis occurred in 22% of Chinese patients. Dual infections with leptospirosis have been reported. Standardized diagnostic tests are being developed and attempts to improve treatment of women and children are being made. Of the numerous tick-borne rickettsioses identified in recent years, African tick-bite fever appears to be of particular importance to travellers. The newly described flea-borne spotted fever caused by Rickettsia felis may be global in distribution. SUMMARY Rash and fever in a returning traveler could be rickettsial and presumptive doxycycline treatment can be curative. Recent research findings raise more questions than answers and should stimulate much needed research.


title: Daily Situation Report on Coronavirus disease (COVID-19) in Iran; March 17, 2020.

text: The main strategy of the Ministry of Health (MOH) for the control of COVID-19 is active case finding using electronic tools, taking advantage of the capacity of the national primary health care (PHC) network, and community engagement. To accomplish this, all people are asked to participate in the screening process using the MOH's self-assessment portal (salamat.gov.ir). Health workers and public volunteers actively contact suspected cases using information recorded in this portal, as well as the electronic health record (SIB portal). They actively evaluate individuals, follow-up them, and provide them with necessary information. Where needed, clients are referred to 16-hour treatment clinics, which are specifically set up for the diagnosis and treatment of the disease, to the hospitals, or will be treated at home. The preliminary results of this national campaign (launched nationwide last week) is provided in this factsheet.


Challenges Facing the Profession 2020: Dental hygiene education in the era of COVID-19.


title: Functional characterization of intracellular and secreted forms of a truncated hepatitis C virus E2 glycoprotein.

text: The E2 protein of hepatitis C virus (HCV) is believed to be a virion surface glycoprotein that is a candidate for inclusion in an antiviral vaccine. A truncated soluble version of E2 has recently been shown to interact with CD81, suggesting that this protein may be a component of the receptor for HCV. When expressed in eukaryotic cells, a significant proportion of E2 forms misfolded aggregates. To analyze the specificity of interaction between E2 and CD81, the aggregated and monomeric forms of a truncated E2 glycoprotein (E2(661)) were separated by high-pressure liquid chromatography and analyzed for CD81 binding. Nonaggregated forms of E2 preferentially bound CD81 and a number of conformation-dependent monoclonal antibodies (MAbs). Furthermore, intracellular forms of E2(661) were found to bind CD81 with greater affinity than the extracellular forms. Intracellular and secreted forms of E2(661) were also found to differ in reactivity with MAbs and human sera, consistent with differences in antigenicity. Together, these data indicate that proper folding of E2 is important for its interaction with CD81 and that modifications of glycans can modulate this interaction. Identification of the biologically active forms of E2 will assist in the future design of vaccines to protect against HCV infection.


Infectious disease epidemiology.


title: Recombinant swine beta interferon protects swine alveolar macrophages and MARC-145 cells from infection with Porcine reproductive and respiratory syndrome virus.

text: Swine beta interferon (swIFN-beta) produced in HEK 293 cells infected with a recombinant, replication-defective human adenovirus 5 (Ad5) encoding the swIFN-beta gene was tested for antiviral activity against Porcine reproductive and respiratory syndrome virus (PRRSV). MARC-145 cells were incubated overnight with dilutions of supernatant fluids from HEK 293 cells infected with Ad5-swIFN-beta or with an Ad5 control virus (Ad5-Blue). Treated cells were infected with PRRSV; MARC-145 cells incubated with Ad5-Blue supernatants developed cytopathic effects (CPE), whereas those incubated with swIFN-beta showed no CPE. To confirm the antiviral activity of swIFN-beta, culture fluids from Ad5-swIFN-beta-infected cells were affinity-purified on a Sepharose-anti-swIFN-beta matrix, and the resulting fractions exhibited antiviral activity upon infection with PRRSV. The antiviral effects were specific, as they were blocked by mAbs against swIFN-beta. Additional cultures of MARC-145 cells treated with swIFN-beta-containing supernatants or affinity-purified swIFN-beta were infected with PRRSV and tested by real-time RT-PCR for viral RNA in culture supernatants at various times post-inoculation. These experiments confirmed the protective effects of swIFN-beta. swIFN-beta was also tested for antiviral activity on porcine alveolar macrophages (PAMs) obtained by bronchoalveolar lavage from PRRSV-negative swine. PAMs were treated with dilutions of swIFN-beta or Ad5-Blue culture fluids, infected with PRRSV and tested for viral RNA by real-time RT-PCR. The viral load data showed a dose-dependent protection in swIFN-beta-treated PAMs, whereas no protection was evident from Ad5-Blue culture fluids. The data demonstrate that swIFN-beta protects both MARC-145 cells and PAMs from PRRSV infection.


title: Cryptogenic organising pneumonia.

text: Organising pneumonia is defined histopathologically by intra-alveolar buds of granulation tissue, consisting of intermixed myofibroblasts and connective tissue. Although nonspecific, this histopathological pattern, together with characteristic clinical and imaging features, defines cryptogenic organising pneumonia when no cause or peculiar underlying context is found. Rapid clinical and imaging improvement is obtained with corticosteroid treatment, but relapses are common after stopping treatment.


title: [Electron microscopy in the diagnosis of enteritis in cats].

text: Over a five-year period (1981-1985) 346 faecal and intestinal samples of cats affected with diarrhoea were studied by electron microscopy. This method revealed the presence of virus in 144 out of 346 (41.6 per cent) samples studied. 117 (81.3 per cent) of these samples contained parvoviruses which were identified by using a specific immune serum (immuno electron microscopy). In addition, rotaviruses (two samples), coronaviruses (thirteen samples), coronavirus-like (ten samples) and picornavirus-like particles (two samples) were detected in the other specimens. The present study shows that electron microscopy is a useful and rapid procedure in the diagnosis of enteritis in cats as well as in other domestic animals.


title: Molecular characterization of the S protein gene of human coronavirus OC43.

text: The gene encoding the spike protein of the OC43 strain of human coronavirus (HCV-OC43) was cloned and sequenced. The complete nucleotide sequence revealed an open reading frame of 4062 nucleotides encoding a protein of 1353 amino acids with a predicted M(r) of 150,078. Structural features include 22 N-glycosylation sites, an N-terminal hydrophobic signal sequence of 17 amino acids, an hydrophilic cysteine-rich sequence of 35 amino acids near the C terminus, and a potential proteolytic cleavage site (RRSR) between amino acid residues 758 and 759, yielding S1 and S2 segments of 84,730 and 65,366 M(r), respectively. The predicted amino acid sequence of the spike protein of HCV-OC43 has 91% identity with that of the Mebus strain of bovine coronavirus, revealing more sequence divergence in the putative bulbous part (S1) than in the predicted stem region (S2).


title: Comparison of electron microscopy, enzyme-linked immunosorbent assay, solid-phase radioimmunoassay, and indirect immunofluorescence for detection of human rotavirus antigen in faeces.

text: Four techniques were compared for their practicability, speed, and sensitivity for the detection of human rotavirus. Radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) were found to be the most sensitive means of identifying rotavirus, and, once processed, up to 40 specimens could be examined daily. Electron microscopy, although less sensitive than these techniques, had the advantage of being able to detect other viral agents present in faecal extracts. Indirect immunofluorescence failed to detect rotavirus as often as the other three methods. In laboratories where routine examination of faecal specimens from patients with gastroenteritis is required, ELISA and RIA are useful alternatives to electron microscopy.


title: Advances in clinical diagnosis and treatment of severe acute respiratory syndrome.

text: It has been proved that severe acute respiratory syndrome (SARS) is caused by SARS-associated coronavirus, a novel coronavirus. SARS originated in Guangdong Province, the People's Republic of China at the end of 2002. At present, it has spread to more than 33 countries or regions all over the world and affected 8 360 people and killed 764 by May 31,2003. Identification of the SARS causative agent and development of a diagnostic test are important. Detecting disease in its early stage, understanding its pathways of transmission and implementing specific prevention measures for the disease are dependent upon swift progress. Due to the efforts of the WHO-led network of laboratories testing for SARS, tests for the novel coronavirus have been developed with unprecedented speed. The genome sequence reveals that this coronavirus is only moderately related to other known coronaviruses. WHO established the definitions of suspected and confirmed and probable cases. But the laboratory tests and definitions are limited. Until now, the primary measures included isolation, ribavirin and corticosteroid therapy, mechanical ventilation, etc. Other therapies such as convalescent plasma are being explored. It is necessary to find more effective therapy. There still are many problems to be solved in the course of conquering SARS.


title: Current treatment for acute viral bronchiolitis in infants.

text: This paper provides an update and critical review of available data on the treatment of acute viral bronchiolitis in previously healthy infants, with special focus on new or promising therapies. The main potential benefits of medical assistance in these patients reside in the careful monitoring of their clinical status, the maintenance of adequate hydration and oxygenation, the preservation of the airway opened and cleared of secretions and the option to perform parental education. There is no convincing evidence that any other form of therapy will reliably provide beneficial effects in infants with bronchiolitis and currently, any treatment beyond supportive care should be prescribed on a case-by-case basis with watchful appraisal of its effects. Therapies such as ribavirin, IFN, vitamin A, antibiotics, mist therapy or anticholinergics, have not demonstrated any measurable clinical effect. Several studies and meta-analyses with beta(2)-agonists and corticosteroids have failed to show any benefit of significant extent, however, physicians keep favouring their use. Presently, adrenaline has received rather consistent support from clinical trials but it is not yet widely prescribed. There are other therapeutic strategies, for instance, heliox, hypertonic saline, noninvasive ventilation, physical therapy techniques, thickened feeds or palivizumab that have shown promising potential benefits, but evidence supporting its use is still limited and further studies should be warranted. In the meantime, infants with acute viral bronchiolitis should be treated following evidence-based clinical practice guidelines, keeping the patient central in the process and being sensitive to social, cultural and familiar influences on their treatment strategy.


title: Predictors of outcome in 216 subjects with ARDS treated with inhaled epoprostenol.

text: BACKGROUND ARDS is an important cause of respiratory failure and continues to be associated with a high mortality rate. Numerous therapeutic interventions have been employed to improve patient outcomes, including inhaled epoprostenol. METHODS We examined subjects with ARDS treated with epoprostenol. We compared hospital survivors with nonsurvivors to identify predictors of mortality. RESULTS Among the cohort (n=216), there were 80 (37%) hospital survivors and 136 (63%) hospital nonsurvivors. Logistic regression revealed 5 variables associated with hospital mortality: trauma as the etiology for ARDS (adjusted odds ratio [AOR] 0.09, 95% CI 0.04–0.22, P=.006), presence of both pulmonary and nonpulmonary sources of sepsis (AOR 3.06, 95% CI 1.98-4.74, P.01), an international normalized ratio of > 1.5 (AOR 3.15, 95% CI 2.19-4.54, P=.002), body mass index (1-unit increments, AOR 0.95, 95% CI 0.936-0.965, P=.001), and an incremental change in PaO2/FIO2 during the first 24 h of treatment with epoprostenol (AOR 0.99, 95% CI 0.988-0.994, P=.002). An analysis for 90-d mortality identified the same predictors, with the addition of cumulative fluid balance during treatment with epoprostenol of > 4 L also being an independent predictor (AOR 2.36, 95% CI 1.66-3.37, P=.02). CONCLUSIONS Although the use of epoprostenol in ARDS remains a therapeutic challenge, we were able to identify predictors of mortality for this important cohort of patients. These predictor variables could be employed in the design of future trials of epoprostenol in ARDS.


Involvement of microtubules and the microtubule-associated protein tau in trafficking of JHM virus and components within neurons.


Antiviral therapy using natural products: introduction.


Canadian surgery and the events of 2003.


Isolation and characterization of the acetylesterase of hemagglutinating encephalomyelitis virus (HEV).


title: A traumatic abdominal wall hernia repair: a laparoscopic approach.

text: BACKGROUND Traumatic abdominal wall hernias from blunt trauma usually occur as a consequence of motor vehicle collisions where the force is tangential, sudden, and severe. Although rare, these hernias can go undetected due to preservation of the skin overlying the hernia defect. Open repairs can be challenging and unsuccessful due to avulsion of muscle directly from the iliac crest, with or without bone loss. A laparoscopic approach to traumatic abdominal wall hernia can aid in the delineation of the hernia and allow for a safe and effective repair. CASE DESCRIPTION A 36-year-old female was admitted to our Level 1 trauma center with a traumatic abdominal wall hernia located in the right flank near the iliac crest after being involved in a high-impact motor vehicle collision. Computed tomography and magnetic resonance imaging of the abdomen revealed the presence of an abdominal wall defect that was unapparent on physical examination. The traumatic abdominal wall hernia in the right flank was successfully repaired laparoscopically. One-year follow-up has shown no sign of recurrence. DISCUSSION A traumatic abdominal wall hernia rarely presents following blunt trauma, but should be suspected following a high-impact motor vehicle collision. Frequently, repair is complicated by the need to have fixation of mesh to bony landmarks (eg, iliac crest). In spite of this challenge, the laparoscopic approach with tension-free mesh repair of a traumatic abdominal wall hernia can be accomplished successfully using an approach similar to that taken for laparoscopic inguinal hernia repair.


title: Single-incision versus conventional laparoscopic cholecystectomy: a case control study.

text: BACKGROUND Single-incision laparoscopic surgery (SILS) is a minimally invasive technique which can be used for treatment of gallbladder disease. To evaluate our initial experience, patients treated with SILS cholecystectomy were compared with a comparable group of patients treated with a conventional four trocart technique (LC). METHODS Between May 2009 and April 2010, 60 SILS cholecystectomies were performed. These patients were matched by Body Mass Index (BMI) with 60 cases of conventional LC. RESULTS The operative time was significantly longer in the SILS group 55 min (range, 25-126 min) compared to 49 min (range, 28-75 min) for the LC group. Excluding the first 15 SILS cases the operative time became comparable to the conventional technique, with a mean operative time of 51 min (range, 25-90 min). No patients were converted to open cholecystectomy. In the SILS group 3 patients developed a wound infection, in the conventional LC group 2. CONCLUSION SILS cholecystectomy seems to be a safe and feasible procedure when performed by an experienced laparoscopic surgeon. Complication rates are comparable to conventional LC. With a learning curve of around 10 to 15 procedures, operative times approach those of conventional LC.


title: [Acetone-celloidin method for the cytological and immunofluorescent study of cell cultures infected with various viruses].

text: A method was worked out for the extraction of a cell monolayer from test tubes with the use of 10 per cent acetone solution of celloidin. This made it possible to carry out parallel cytologic and immunofluorescent studies of cultures infected with various viruses. In contrast to the alcohol-ether celloidin used in histology the use of the acetone solution of celloidin, suggested by us, preserved to a larger extent the viruses of different structure and composition, and, therefore, was shown to be more suitable in immunofluorescent investigations. The method was employed for the cytologic and the immunofluorescent identification of newly isolated virus agents. It was also used in the controlled replication of noncytopathogenic and slightly cytopathogenic strains of bovine rotaviruses, corona viruses, respiratory-syncytial virus, pestivirus, and others that required roller cultivation. The celloidin method makes it possible to obtain from one to four preparations with material of the cell monolayer of each test tube, and renders it unnecessary to maintain permanently cultures on glass lamellas.


title: Retrospective evidence that the MHC (B haplotype) of chickens influences genetic resistance to attenuated infectious bronchitis vaccine strains in chickens.

text: Infectious bronchitis is a respiratory disease of chickens that is caused by the coronavirus infectious bronchitis virus (IBV). Virtually all broiler and layer breeder flocks are routinely vaccinated against IBV. Two hatches of 1-day-old chicks from four lines were mistakenly vaccinated for infectious bronchitis using a moderately attenuated vaccine designed for chicks of an older age. The vaccination resulted in high mortality, and chicks from three of four lines died with signs typical of infectious bronchitis. The mortality that occurred using this less-attenuated vaccine was significantly influenced by the genetic line, and the MHC (B) haplotype in chickens of three B congenic lines. B congenic chickens possessing the B*15 haplotype were resistant in contrast to chickens possessing the B*13 or B*21 haplotypes. Chicks from two further hatches of the four lines were vaccinated appropriately with a more attenuated IBV vaccine, and only limited chick mortality was seen. These retrospective data from two repeated hatches confirm earlier data indicating chicken genes influence resistance to IBV, and indicate for the first time that genes tightly linked to the B haplotype are relevant in resistance to IBV. Due to extenuating circumstances it was not possible to verify results with chicks from F2 matings. Factors that may enhance definition of the role of the B haplotype in immune response to IBV, and the desirability for further analysis of a B haplotype-linked influence on immunity to IBV are discussed.


title: Immunoglobulin derived from bovine plasma as a replacement for colostrum in newborn lambs.

text: Newborn lambs (n = 45) at the Agricultural Research Institute of Northern Ireland were fed either 50 grams of commercial lamb milk replacer or 50 grams of commercial colostrum replacer (bovine origin) in 200 ml of water four times during the first 24 hours of life or were given ad libitum access to the ewe. Total plasma protein at 24 hours of age was highest in lambs allowed to suckle the ewe (76.9 g/L). However, by 14 days of age, there were no differences in plasma protein levels among the three treatments. Bovine IgG was measured in lambs fed colostrum replacer and ovine IgG was measured in other lambs. Mean plasma IgG concentrations at 24 hours of age were 0.7 (milk replacer), 18.0 (colostrum replacer), and 26.6 (dam's milk) g/L. Bovine IgG administered orally to newborn lambs was adequately absorbed, and circulating IgG concentrations were sufficiently maintained throughout this study.


title: [Cell entry mechanism of coronaviruses: implication in their pathogenesis].

text: Coronaviruses infect many species of animals, including humans. Among them, murine coronavirus, mouse hepatitis virus (MHV) has been well studied as a model of human diseases, such as hepatitis and demyelinating disease. An agent causing severe acute respiratory disease (SARS), SARS coronavirus (SARS-CoV), is a newcomer in this genus, however, it is now one of the most studied coronaviruses due to its medical impact. The receptors of those two viruses have been identified and their cell entry mechanism has been actively investigated. Recently, SARS-CoV cell entry mechanism is shown to be different from that of other enveloped viruses, including MHV. In this review, cell entry mechanism of those two viruses is described, stressing on the difference and similarity found between those two viruses.


title: Comparative viral frequency in Mexican children under 5 years of age with and without upper respiratory symptoms.

text: In Mexico, there is a lack of up-to-date published data that show viruses to be the main cause of acute respiratory infection (ARI). The objective of this study was to estimate the comparative viral frequency between children under the age of 5 years with and without ARI (n = 179 in each group) in a suburban community (Nezahualcóyotl City). A nasopharyngeal sample was collected for viral culture and identification was carried out by indirect immunofluorescence (IIF) using mAbs. There were no sex differences between the two groups. Children under 1 year of age with ARI showed a higher frequency (56 %) of viral infections; this was statistically significant (P < 0.05) when compared with the same age group in ARI-free children (17 %). Respiratory syncytial virus (RSV) was the most prevalent type of virus isolated from both groups (38 vs 18 %). A statistically significantly higher number of subjects with ARI (33/179) than without (12/179) were infected with RSV (P < 0.003). Prevalences of four other viruses studied were similar in the two groups. The highest viral incidence of ARI in children was detected in the winter-spring seasonal period.


title: Characterization of the coronavirus mouse hepatitis virus strain A59 small membrane protein E.

text: The small envelope (E) protein has recently been shown to play an essential role in the assembly of coronaviruses. Expression studies revealed that for formation of the viral envelope, actually only the E protein and the membrane (M) protein are required. Since little is known about this generally low-abundance virion component, we have characterized the E protein of mouse hepatitis virus strain A59 (MHV-A59), an 83-residue polypeptide. Using an antiserum to the hydrophilic carboxy terminus of this otherwise hydrophobic protein, we found that the E protein was synthesized in infected cells with similar kinetics as the other viral structural proteins. The protein appeared to be quite stable both during infection and when expressed individually using a vaccinia virus expression system. Consistent with the lack of a predicted cleavage site, the protein was found to become integrated in membranes without involvement of a cleaved signal peptide, nor were any other modifications of the polypeptide observed. Immunofluorescence analysis of cells expressing the E protein demonstrated that the hydrophilic tail is exposed on the cytoplasmic side. Accordingly, this domain of the protein could not be detected on the outside of virions but appeared to be inside, where it was protected from proteolytic degradation. The results lead to a topological model in which the polypeptide is buried within the membrane, spanning the lipid bilayer once, possibly twice, and exposing only its carboxy-terminal domain. Finally, electron microscopic studies demonstrated that expression of the E protein in cells induced the formation of characteristic membrane structures also observed in MHV-A59-infected cells, apparently consisting of masses of tubular, smooth, convoluted membranes. As judged by their colabeling with antibodies to E and to Rab-1, a marker for the intermediate compartment and endoplasmic reticulum, the E protein accumulates in and induces curvature into these pre-Golgi membranes where coronaviruses have been shown earlier to assemble by budding.


title: Distinct signature type I interferon responses are determined by the infecting virus and the target cell.

text: BACKGROUND Type I interferons (IFN) include multiple IFN-alpha subtypes which exhibit considerable amino acid identity and activate the same cell-surface receptor. The promoter regions of the IFN-alpha genes, however, have different transcription factor binding sites, implying differential transcriptional activation. Evolutionary conservation of multiple subtypes may have resulted from external pressures associated with the crucial nature of an IFN response, namely that different viruses that are tropic for different target tissues determine the nature and extent of an IFN response, specifically the IFN-alpha subtype profile. METHODS Studies were undertaken to examine inducible IFN gene expression profiles in response to infection with single-stranded RNA viruses: Sendai virus (SeV), murine hepatitis virus (MHV-1) and coxsackie virus B3 (CVB3). RESULTS In vitro, distinct signature profiles of SeV and MHV-1-inducible gene expression for IFN-alpha2, IFN-alpha4 and IFN-alpha5 subtypes in L2 and L929 mouse fibroblast cells, in relation to the extent and kinetics of their induction, were identified. In vivo, whereas A/J mice are highly permissive for both MHV-1 and CVB3 infections and mount a poor IFN response, C57B1/6 mice are relatively resistant to both virus infections and mount a vigorous IFN response. CONCLUSIONS These data suggest that the infecting virus and the target cell type dictate the extent and signature of inducible type I IFN gene expression. The extent of IFN response to viral infection influences the subsequent biological outcome: a robust IFN response prescribes a level of resistance, whereas a poor IFN response contributes towards a permissive phenotype for infection.


title: Pandemic influenza and the hospitalist: apocalypse when?

text: Beginning with a cluster of human cases in Hong Kong in 1997, avian influenza (H5N1) has spread progressively through, and beyond, Asia in poultry and other birds; and has resulted in sporadic cases of human disease associated with high mortality. The potential for H5N1 influenza to cause a pandemic of human disease continues to be the subject of intense scrutiny by both the media and the scientific community. While the likelihood of such a prospect is uncertain, the inevitability of future pandemics of influenza is clear. Planning for the eventuality of a virulent influenza pandemic at the local, national and global level is critical to limiting the mortality and morbidity of such an occurrence. Hospitalists have a key role to play in institutional efforts to prepare for a influenza pandemic, and should be aware of lessons that my be applied from both the response to Hurricane Katrina, as well as the severe acute respiratory syndrome (SARS) epidemic.


title: Genetic recombination of poliovirus in vitro and in vivo: temperature-dependent alteration of crossover sites.

text: Genetic recombination that occurs with high frequency during poliovirus genome replication is a process whose molecular mechanism is poorly understood. Studies of genetic recombination in a cell-free system in vitro and in infected tissue culture cells in vivo have led to the unexpected observation that temperature strongly influences the loci at which cross-over between the two recombining RNA strands occurs. Specifically, cross-over between two genetically marked RNA strands in vitro and in vivo at 34 degrees C occurred over a wide range of the genome. In contrast, recombination in vivo at 37 and 40 degrees C yielded cross-over patterns that had shifted dramatically to a region encoding nonstructural proteins. Preferential selection of recombinants at 37 and 40 degrees C was ruled out by analyses of the growth kinetics of the recombinants. During the studies of recombination in the cell-free system we found that there is a direct correlation between the ability of a poliovirus RNA molecule to replicate in the cell-free system and its capacity to complement de novo virus synthesis programmed by another viral RNA.


title: Efficacy of a deletion mutant bovine herpesvirus-1 (BHV-1) vaccine that allows serologic differentiation of vaccinated from naturally infected animals.

text: Fifteen bovine herpesvirus-1 (BHV-1)-negative calves were vaccinated intramuscularly with 10(7.4) plaque-forming units of a double-deletion BHV-1 mutant (IBRV(NG)dltkdlgIII), and 6 remained as nonvaccinated controls. Thirty days after vaccination, the animals were challenged by nasal instillation of 10(8.2) CCID50 of a virulent BHV-1 strain (Cooper). The vaccinated calves were protected against wildtype virus challenge as demonstrated by clinical evaluation. Most of the vaccinates developed only a mild rhinitis (lasting an average of 6.5 days) with almost no systemic symptoms, whereas the controls developed a serious illness characterized by rhinitis (mean = 11.5 days), conjunctivitis, hyperthermia, apathy, loss of appetite, and dyspnea. The vaccinates also shed significantly less virus and for a shorter period of time (mean = 5.5 days) than the controls (mean = 9 days). Thirty days after vaccination, the vaccinates were negative in an anti-gIII specific blocking enzyme-linked immunosorbent assay (ELISA), despite the fact that most of them had developed neutralizing antibodies (serum neutralization titers ranging from 1:2 to 1:16). Seroconversion to gIII was detected as early as 7 days postinfection (dpi). Fourteen days after the challenge, all the animals exposed to wildtype BHV-1 had developed anti-gIII antibodies and were positive in this differential serologic test. Six controls plus 8 vaccinates kept in isolation were still positive to gIII when tested at 75 dpi. The use of the IBRV(NG)dltkdlgIII strain in conjunction with an anti-gIII specific blocking ELISA kit represents a powerful tool for BHV-1 control/eradication programs.


title: A case of giant mucinous cystic neoplasm of the pancreas resected with laparoscopic surgery.

text: We report a case of giant mucinous cystic neoplasm (MCN) of the pancreas that was successfully treated with laparoscopic surgery. A 29-year-old woman was admitted to our hospital for evaluation of an abdominal tumor that had been detected during a routine medical examination. The tumor was diagnosed as an MCN of the pancreas. We performed laparoscopic distal pancreatectomy and splenectomy. The patient could begin oral feeding 3 days after the operation. No complications occurred either during or after surgery. The outcome in this patient was excellent, with cosmetically acceptable small scars in the skin. Therefore, laparoscopic distal pancreatectomy may become a therapeutic option for patients with large MCNs of the pancreas.


title: Anogenital neoplasia in AIDS.

text: PURPOSE OF REVIEW We review the recent literature on anogenital neoplasms in AIDS, with emphasis on cancers associated with HPV infection. Immune reactivity to HPV as well as novel immunotherapeutic and preventative strategies are discussed. RECENT FINDINGS Many AIDS-associated neoplasms are associated with HPV infection. Whether cervical cancer is truly an AIDS-associated neoplasm has recently been questioned, while the association of anal cancer with AIDS in both males and females is more convincing. Recent reports cast doubt on the efficacy of HAART therapy for HPV-induced anogenital neoplasms, despite efficacy in improving disease caused by other infectious agents. We include here new data on humoral and cellular immune responses to HPV. VLP serology has been reported to be associated with outcome of cervical cancer. VLP seropositivity has been reported to be a favorable prognostic sign in women with HPV 16 positive cervical carcinoma. Several investigators have questioned the immunogenicity of the oncogenic HPV type 16 compared with other HPV types. It has recently been found that in HIV-infected patients, lymphoproliferative cellular immune responses (CMI) to HPV 16 peptides are not associated with CD4 counts, whereas responses to recall antigens and mitogens are associated with CD4 counts. CD4 + T cells may not be responsible for protective cellular immune responses to HPV. VLP serology and CMI responses may be the future intermediate surrogate biomarkers for HPV-associated anogenital neoplasia trials. Several new therapeutic vaccine strategies for management of HPV-induced neoplasia are reviewed. SUMMARY Most anogenital neoplasms occurring with increased frequency in patients with HIV/AIDS are associated with HPV infections. Current treatment strategies are not effective in clearing anogenital HPV infection and need improvement. Immunotherapy with novel vaccines will provide both prevention and therapy for these common malignancies.


title: [Comments on treatment of severe acute respiratory syndrome by integrated traditional Chinese and western medicine].

text: There are obvious advantages of the integrated traditional Chinese and western medicine on the treatment of severe acute respiratory syndrome (SARS). Combining the ancient and present literature with the special epidemic patterns, pathological changes and clinical symptoms of SARS, the paper discussed the methods of application of the integrated traditional Chinese and western medicine on the treatment of SARS, and some matters needing attention in clinic.


title: Photolabeling probes of ribavirin and EICAR.

text: Ribavirin, the only small molecule available so far for treating hepatitis C virus infection, was recently used in an emergency context to treat patients with severe acute respiratory syndrome in the early stages of the disease. EICAR, one of the most potent congeners of ribavirin, has 10 to 100 times greater antiviral potency than ribavirin. The mechanisms underlying the antiviral effects of ribavirin and EICAR have not yet been definitely elucidated, but they seem to be similar. In order to study the mechanisms responsible for their antiviral effects using a photolabeling approach, we have developed photolabeling probes of ribavirin and EICAR, in which an azido group was introduced into the pseudobases of triazole and imidazole, respectively. The ribavirin photoprobes were obtained by directly coupling the azidotriazole to the protected ribose sugar, while the EICAR probe was prepared by diazotizing AICAR and subsequently substituting with NaN3. All these probes showed a fast, clear-cut photochemical reaction, which suggests that they are promising tools for use in photolabeling studies.


title: Paramyxovirus infection in pigs with interstitial pneumonia and encephalitis in the United States.

text: In the last few years, newly recognized paramyxoviruses have been associated with severe disease in several animal species, including swine, as well as in human beings. Recently, a paramyxovirus was isolated from a swine herd in the northcentral United States that experienced an epizootic of respiratory and central nervous system disease. Affected pigs had interstitial pneumonia with necrotizing bronchiolitis and encephalitis characterized by lymphocytic perivasculitis and diffuse gliosis. Germ-free pigs inoculated with this isolate developed mild clinical illness and similar but less severe histopathologic lesions in lungs and brain.


title: Molecular mechanism of membrane protein integration into the endoplasmic reticulum.

text: As proteins are integrated into the membrane of the endoplasmic reticulum, some hydrophilic polypeptide segments are transported through the translocation channel, others remain in the cytosol, and hydrophobic transmembrane sequences are released into the lipid phase. We have addressed the molecular mechanism by which these events occur. We demonstrate that both the lumenal and the cytosolic domains of a membrane protein are synthesized while the ribosome is membrane bound, so that even cytosolic domains come in contact with the translocation channel. We also find that, before translation of the protein is terminated, transmembrane sequences can laterally exit the translocation channel and enter the lipid environment. These results have significant implications for the folding and assembly of membrane proteins.


title: Hepatitis B vaccination and first central nervous system demyelinating event: a case-control study.

text: OBJECTIVE To investigate the relationship between hepatitis B (HB) vaccination and a first central nervous system (CNS) demyelinating event in adults. METHODS In 1998, we conducted a multicentre, hospital-based case-control study which enrolled 402 cases of first CNS demyelinating event occurring between 1994 and 1995 and 722 controls matched for centre, age, sex and date of admission. An independent expert committee validated the diagnoses of cases and controls. Data on vaccinations were obtained from a standardized phone interview. Forty percent of eligible cases and 50% of eligible controls could not be localized or were excluded because they did not satisfy inclusion or matching criteria. RESULTS Conditional logistic regression performed on 236 and 355 matched controls showed that adjusted odds ratios for the first CNS demyelinating event within 2 months following an injection of HB vaccine were 1.8 [95% confidence interval (CI), 0.7-4.6] in the whole group and 1.4 (95% CI, 0.4-4.5) in the subgroup of cases (n = 152) and controls (n = 253) referring to vaccination certificates during the phone interview. Restricting the analyses to the cases with definite or probable multiple sclerosis, these odds ratios were 2.0 (95% CI, 0.8-5.4) and 1.6 (95% CI, 0.4-5.6), respectively. Odds ratios tend towards 1 for a longer interval between HB vaccine and demyelinating event. CONCLUSIONS This study was sufficiently powerful to rule out a strong association between HB vaccine exposure and a subsequent demyelinating event. However, it could not provide a clear indication of a moderately increased risk of a CNS demyelinating event shortly after HB vaccination in adults.


title: [Epidemiological features of severe acute respiratory syndrome in Beijing].

text: OBJECTIVE To describe the epidemiologic features of severe acute respiratory syndrome (SARS) in Beijing. METHOD Database of the 2 521 probable cases of SARS in Beijing Center for Disease Prevention and Control was used. RESULTS The course of SARS epidemic in Beijing could be divided into five phases: import and spreading-from 1 to 31 in March, rising-from April 1 to April 15, peak-from April 16 to May 4, declining-from 5 to 18 in May, terminating-from 19 to 28 in May. The proportions of portable cases of SARS in each phase were 2.7%, 13.6%, 71.0%, 11.6% and 1.1%, respectively. Totally, 2 521 portable cases were diagnosed and verified according to the diagnostic criteria of SARS issued by the Ministry of Health. Among them, 192 died from SARS. The incidence and mortality rates of SARS were 18.57 per 100,000 and 1.41 per 100,000 with the fatality of 7.6%. The ratio of male to female with SARS was 1:0.97. The highest incidence rate of SARS was in the group of 20 - 29 years (30.85 per 100,000), and the lowest was in the group of 0 - 14 years (2.54 per 100,000). People aged 20 - 49 accounted for 72.3% of all SARS cases. The incidence rates in urban, suburb and far-suburb were 32.25/100,000, 20.57/100,000 and 8.90/100,000, respectively, decreasing according to the population density. Health care providers (17.3%), staff (12.9%), retirees (11.4%), workers (9.7%) and house-hold unemployees (8.8%) appeared to be at the five top risk populations being infected. The fatality increased significantly with age. CONCLUSION Beijing was the most severe epidemic region of SARS in the world, but the fatality was the lowest.


title: Modulation of the suppressive effect of corticosterone on adult rat hippocampal cell proliferation by paroxetine.

text: OBJECTIVE The literature has shown that cognitive and emotional changes may occur after chronic treatment with glucocorticoids. This might be caused by the suppressive effect of glucocorticoids on hippocampal neurogenesis and cell proliferation. Paroxetine, a selective serotonin reuptake transporter, is a commonly used antidepressant for alleviation of signs and symptoms of clinical depression. It was discovered to promote hippocampal neurogenesis in the past few years and we wanted to investigate its interaction with glucocorticoid in this study. METHODS Adult rats were given vehicle, corticosterone, paroxetine, or both corticosterone and paroxetine for 14 d. Cell proliferation in the dentate gyrus was quantified using 5-bromo-2-deoxyuridine (BrdU) immunohistochemistry. RESULTS The corticosterone treatment suppressed while paroxetine treatment increased hippocampal cell proliferation. More importantly, paroxetine treatment could reverse the suppressive effect of corticosterone on hippocampal cell proliferation. CONCLUSION This may have clinic application in preventing hippocampal damage after glucocorticoid treatment.


title: Sequence of the spike protein of the porcine epidemic diarrhoea virus.

text: The complete sequence of the spike (S) gene of the Br1/87 isolate of porcine epidemic diarrhoea virus (PEDV) was determined from cDNA clones. The predicted polypeptide was 1383 amino acids long, contained 29 potential N-linked glycosylation sites and showed structural features similar to those of the coronavirus spike protein. The PEDV S protein, like that of the members of the transmissible gastroenteritis virus (TGEV)-related subset, lacks a proteolytic site to yield cleaved amino and carboxy subunits S1 and S2. Viral polypeptide species of the expected M(r), i.e. 170K/190K, were observed in PEDV-infected cells. Sequence comparison confirmed that, within the subset, PEDV was most closely related to the human respiratory coronavirus HCV 229E. However, PEDV S protein has an additional 250 residue N-terminal domain which is absent from HCV 229E and porcine respiratory coronavirus, the respiratory variant of TGEV. Alignment of the S1 regions revealed a second domain of about 90 residues with increased sequence divergence which might possibly express virus-specific determinants.


title: The causes and diagnosis of influenza-like illness.

text: BACKGROUND Influenza and other respiratory viruses circulate between spring and autumn in temperate climates and all year in tropical climates. These viruses cause symptoms often referred to as influenza-like illness (ILI), but are not generally distinguishable on clinical grounds alone. OBJECTIVE This article provides a brief review of the surveillance, viral causes and current diagnostic methods used to identify viruses causing ILI. DISCUSSION Influenza-like illness surveillance with laboratory support, conducted in most Australian stats and territories, aims to define the impact of influenza seasons in the community and provide virus strains that may be used in future vaccine formulations. Surveillance may also be useful in the early stages of an influenza pandemic. In addition to influenza, viruses known to cause ILI include respiratory syncytial virus, rhinovirus, adenovirus, parainfluenza viruses, human coronaviruses (including the virus that causes severe acute respiratory syndrome) and the recently recognised human metapneumovirus. Polymerase chain reaction assays are the most common diagnostic tests now used for the differential diagnosis of ILI.


Medical considerations for supporting elite athletes during the post-peak phase of the New Zealand COVID-19 pandemic: a New Zealand sporting code consensus.


title: [Viral zoonoses].

text: In different areas of the World, whether in industrialized or developing countries, zoonoses represent an emerging Public Health concern either as known agents appear in areas or species in which they had not been reported or as new pathogens parasite Human as consequence of successful host switching. Epidemiological data outlined that vectorborne viral zoonoses are of particular relevance. This paper review the changes in epidemiology of bat rabies and arbovirus encephalitis.


title: Case management of ill travelers with infectious diseases.

text: Recent increase of international travelers raises awareness of travel-related infections. Travelers' diarrhea is the most common and caused by diarrheagenic E. coli and other patho- gens. Undifferentiated fever challenges physicians to correctly diagnose and includes ma- laria, dengue, and enteric fever. The pre-test probability for a disease which causes undiffer- entiated fever largely depends on the travel destination. Malaria is most common in travelers returning from sub-Saharan Africa while dengue is most common those returning from Southeast Asia and Latin America. Screening for asymptomatic travelers is not generally rec- ommended, but for long-term travelers and immigrants. Following outbreaks of highly infectious diseases in the past decades, hospital prepared- ness for travel-related infections focuses on infection prevention and control. Emerging viral pneumonias such as Middle East respiratory syndrome (MERS) and avian influenza H7N9 are major concerns. As seen in the largest outbreak of Ebola virus disease in West Africa, imported viral hemorrhagic fever such as Lassa fever is another concern. Since multidrug resistant bacteria silently spread globally by infected travelers, it would be of help to install appropriate isolation precaution for travelers with history of hospital admission abroad.


title: Ostrich diseases.

text: Scientific knowledge of ostrich diseases is incomplete and very fragmented, with specific details on technical aspects of diagnostic and/or screening tests completely absent in most cases. Salmonella Typhimurium is common in multispecies collections and causes mortality in chicks younger than three months on commercial farms, but is rarely found in chicks older than six months, or slaughter birds of twelve to fourteen months in southern Africa. Campylobacter jejuni and Chlamydia psittaci are occasionally reported, mainly in young ostriches, but both remain a diagnostic challenge. Crimean-Congo haemorrhagic fever is transmitted to domestic animals including ostriches, principally by ticks of the genus Hyalomma. In the ostrich, the disease causes no clinical symptoms during a viraemia of approximately four days. Spongiform encephalopathy has not been reliably reported in ostriches, while anthrax has occurred rarely in modern times but was reportedly an important cause of death approximately 100 years ago in South Africa. Salmonella Gallinarum and S. Pullorum are unknown in ostriches. Pasteurella multocida occurs but is easily contained with antibiotics. Mycoplasma spp. are regularly found in an upper respiratory disease syndrome complicated by opportunistic bacterial pathogens. Ostriches of all ages are susceptible to challenge by velogenic Newcastle disease virus (NDV), but standard inactivated La Sota poultry vaccines can stimulate protective immunity lasting over six months. The viraemic period in vaccinated slaughter ostriches is between nine and eleven days and there are no indications of a carrier state or presence of the virus in the meat or any other tissues after this period, with peak immunoglobulin G response reached on day fourteen post infection. Haemagglutination inhibition tests are significantly less sensitive and less specific than enzyme-linked immunosorbent assays. Cloacal and choanal swabs used for direct virological screening in clinically affected cases (field and experimental) could not detect NDV. All avian influenza isolates reported from ostriches have been non-pathogenic to poultry, even the H5 and H7 subtypes. Some of the latter have been associated with mortality of ostrich chicks in localised outbreaks during periods of inclement weather and with significant wild bird (waterfowl) contact. Borna disease causes a nervous syndrome in ostrich chicks, but to date, has only been reported in Israel. Eastern and Western equine encephalomyelitides cause fatal disease in ostriches and other ratites, with mortality ranging from less than 20% to over 80% in affected flocks. These diseases are present in North, Central and South America where the associated ornithophilic mosquito vectors occur. Equine and human vaccines are apparently safe and efficacious in ratites. Wesselsbron disease, infectious bursal disease (type 2), adenovirus and coronavirus infections have been reported from ostriches but the significance of these diseases is unclear. Due to the paucity of data regarding ostrich diseases and the unvalidated state of most poultry tests in this unique group of birds, strict observation of a pre-slaughter quarantine of thirty days is strongly advised, whilst live exports and fertile eggs should be screened through the additional use of sentinel chickens and/or young ostriches.


title: [Simultaneous multiple assay (Luminex xTAG respiratory viral panel FAST assay) efficacy in human respiratory virus detection].

text: Luminex xTAG respiratory viral panel FAST (RVP FAST) assay detects 17 human respiratory virus strains per measurement. Studying RVP FAST efficacy in detecting respiratory viruses in 67 aspirate samples from the nasal cavities of children with acute respiratory infection, we compared RVP FAST results to those of conventional nucleic acid amplification tests (NAT), e.g., real-time PCR, targeting 8 strains. RVP FAST assay detected 13 strains (98 isolates) in 59 of 67 samples. Of these, 8--influenza virus (Inf.V)-AH1, Inf. V-AH3, novel Inf.V-AH1, and Inf.V-B, and adenovirus, RS virus, metapneumovirus, and bocavirus--were compared to NAT results. RVP FAST showed higher sensitivity (83.3-100%) and specificity (98.2-100%) than NAT. RVP FAST also detected coronavirus (CoV) 229E, OC43, NL63, and HKU1 from 10 virus strain samples and enterovirus and/or rhinovirus from 35. RVP FAST assay thus comprehensively detects clinically important viruses in a single measurement, making RVP FAST assay useful in detecting causative respiratory tract viruses.


Extensively overlapping reading frames in a second mammalian gene.


title: Increased epitope-specific CD8+ T cells prevent murine coronavirus spread to the spinal cord and subsequent demyelination.

text: CD8+ T cells are important for clearance of neurotropic mouse hepatitis virus (MHV) strain A59, although their possible role in A59-induced demyelination is not well understood. We developed an adoptive-transfer model to more clearly elucidate the role of virus-specific CD8+ T cells during the acute and chronic phases of infection with A59 that is described as follows. C57BL/6 mice were infected with a recombinant A59 virus expressing the gp33 epitope, an H-2Db-restricted CD8+ T-cell epitope encoded in the glycoprotein of lymphocytic choriomeningitis virus, as a fusion with the enhanced green fluorescent protein (RA59-gfp/gp33). P14 splenocytes (transgenic for a T-cell receptor specific for the gp33 epitope) were transferred at different times pre- and postinfection (p.i.). Adoptive transfer of P14 splenocytes 1 day prior to infection with RA59-gfp/gp33, but not control virus lacking the gp33 epitope, RA59-gfp, reduced weight loss and viral replication and spread in the brain and to the spinal cord. Furthermore, demyelination was significantly reduced compared to that in nonrecipients. However, when P14 cells were transferred on day 3 or 5 p.i., no difference in acute or chronic disease was observed compared to that in nonrecipients. Protection in mice receiving P14 splenocytes prior to infection correlated with a robust gp33-specific immune response that was not observed in mice receiving the later transfers. Thus, an early robust CD8+ T-cell response was necessary to reduce virus replication and spread, specifically to the spinal cord, which protected against demyelination in the chronic phase of the disease.