title: [Perinatal and neonatal management plan for prevention and control of SARS-CoV-2 infection (2nd Edition)].

text: Since late December 2019, SARS-CoV-2 infection has spread to all parts of the country and overseas, and the outbreak continues. At the end of January 2020, the Working Group for the Prevention and Control of Neonatal SARS-CoV-2 Infection in the Perinatal Period of the Editorial Committee of Chinese Journal of Contemporary Pediatrics worked out the perinatal and neonatal management plan for prevention and control of SARS-CoV-2 infection (1st Edition). This plan has been verified by clinical practice for 3 weeks. With the further understanding of SARS-CoV-2 infection, especially the emergence of SARS-CoV-2 infection cases in pregnant women and neonates, it is necessary to update the first edition of the management plan so as to offer a better guide on clinical practice. Therefore, the Working Group has worked out the second edition of the management plan.


Coronavirus?


title: Hydrolysis of angiotensin peptides by human angiotensin I-converting enzyme and the resensitization of B2 kinin receptors.

text: We measured the cleavage of angiotensin I (Ang I) metabolites by angiotensin I-converting enzyme (ACE) in cultured cells and examined how they augment actions of bradykinin B2 receptor agonists. Monolayers of Chinese hamster ovary cells transfected to stably express human ACE and bradykinin B2 receptors coupled to green fluorescent protein (B2GFP) or to express only coupled B2GFP receptors. We used 2 ACE-resistant bradykinin analogues to activate the B2 receptors. We used high-performance liquid chromatography to analyze the peptides cleaved by ACE on cell monolayers and found that Ang 1-9 was hydrolyzed 18x slower than Ang I and &30% slower than Ang 1-7. Ang 1-7 was cleaved to Ang 1-5. Although micromol/L concentrations of slowly cleaved substrates Ang 1-7 and Ang 1-9 inhibit ACE, they resensitize the desensitized B2GFP receptors in nmol/L concentration, independent of ACE inhibition. This is reflected by release of arachidonic acid through a mechanism involving cross-talk between ACE and B2 receptors. When ACE was not expressed, the Ang 1-9, Ang 1-7 peptides were inactive. Inhibitors of protein kinase C-alpha, phosphatases and Tyr-kinase blocked this resensitization activity, but not basal B2 activation by bradykinin. Ang 1-9 and Ang 1-7 enhance bradykinin activity, probably by acting as endogenous allosteric modifiers of the ACE and B2 receptor complex. Consequently, when ACE inhibitors block conversion of Ang I, other enzymes can still release Ang I metabolites to enhance the efficacy of ACE inhibitors.


title: Avian influenza: preparing for a pandemic.

text: Avian influenza A (H5N1) first emerged as a global public health threat in 1997 when it caused a major human outbreak in Hong Kong. Endemic in waterfowl and highly virulent in poultry, H5N1 is capable of incidentally infecting humans and other mammals. Although H5N1 is not yet capable of efficient human-to-human transmission, the protean nature of its genome could transform it into the source of the next human influenza pandemic. In the spring of 2006, migrating birds spread the virus from Asia to Europe and Africa. Preparing for a new influenza pandemic involves increasing global influenza surveillance and developing practical strategies for containing outbreaks at the source. Prompt case recognition, isolation, and treatment will be crucial for disease control. Pharmacologic interventions will focus on streamlining the production of vaccine, extending vaccine supplies, stockpiling antiviral drugs such as oseltamivir, and distributing these agents in a timely manner to persons who have the most need. Nonpharmacologic measures will include the use of masks, social distancing, quarantine, travel restrictions, and increasing the emergency capacity of health care systems.


title: Use of Remdesivir in the Management of COVID-19: A Systematic Review on Current Evidences.

text: The rapid progression of corona virus disease in 2019 (COVID-19) pandemic has become an unprecedented global concern. This systemic review aimed at evaluating the available evidence on efficacy, safety to identify any promising role for compassionate use of remdesivir in patient suffered for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) as re-purposeful use. We searched PubMed, EMBASE, Cochrane Library for randomized controlled trials (RCTs), prospective case series studies and case reports that evaluated use of remdesivir in COVID-19. The outcomes were mortality, recovery rate, length of hospital stay and clinical outcome. Though the drug remdesivir (RDV) is not approved by the FDA, still the "Emergency Use Authorization" (EUA) for compassionate use in severe cases is endorsed. After vigorous searching, screening and sorting of completed and published scientific evidences in electronic database, there were only 2 randomized control trial (RCT), 2 uncontrolled trials found until April 2020. We also included 3 published case reports to analyze the validity use of RDV because of the scarcity of evidence based reports. Remdesivir was thought to be one of the promising options for treating the patients of COVID-19 based on few laboratory experiments and reports from some compassionate use and case reports. The safety and efficacy of this drug in COVID-19 cases require high-quality evidence from well-designed and adequately-powered clinical trials with proper sample size for precise decision.


title: Comparison of Early Results and Complications between Multi-and Single-Port Sleeve Gastrectomy: A Randomized Clinical Study.

text: BACKGROUND In recent years, laparoscopic sleeve gastrectomy (LSG) has become more acceptable for obese patients. Single-port sleeve gastrectomy (SPSG) is more popular since each abdominal incision carries the risk of bleeding, hernia, and internal organ injury as well as exponentially affecting cosmesis. This cross-sectional study aimed at comparing multi-port sleeve gastrectomy (MPSG) and SPSG in terms of their early results and complications. METHODS Out of129 obese patients candidated for LSG, 102 patients were assigned to 2 groups of SPSG and MPSG. Complications and demographic data such as body mass index (BMI), age, gender, operation time, and hospital stay were measured. All surgeries were carried out between2013 and 2015 in Shiraz, Iran. Data analysis was performed using SPSS, version 16 for Windows (SPSS Inc., Chicago, IL). The continuous and categorical variables were compared using the Student t-test and the Chi-square test or the Fisher exact test, respectively. RESULTS The patients' data from both groups were similar in terms of age, intraoperative and postoperative bleeding volume, and length of hospital stay. Mean BMI was 42.8±0.7 in the SPSG group and 45.3±1.2 in the MPSG group. Duration of surgery was significantly lower in the SPSG group (P<0.001). Only 1 patient from the SPSG group and 5 patients from the MPSG group had bleeding as an early complication. CONCLUSION The differences in each complication between the groups were not statistically significant. SPSG seems to be safe and is the same as MPSG in terms of major postoperative complications. TRIAL REGISTRATION NUMBER IRCT201512229936N12 The abstract was presented in the 4th International Congress of Minimally Invasive Surgery, Iran, as a poster and published in Iran J Med Sci Journal as a supplement (May 2015; Vol. 40, No. 3).


title: Clinical features and complete genome characterization of a distinct human rhinovirus (HRV) genetic cluster, probably representing a previously undetected HRV species, HRV-C, associated with acute respiratory illness in children.

text: Although human rhinoviruses (HRVs) are common causes of respiratory illness, their molecular epidemiology has been poorly investigated. Despite the recent findings of new HRV genotypes, their clinical disease spectrum and phylogenetic positions were not fully understood. In this study, 203 prospectively collected nasopharyngeal aspirates (NPAs), negative for common respiratory viruses (83 were human bocavirus [HBoV] positive and 120 HBoV negative), from hospitalized children during a 1-year period were subjected to reverse transcription-PCR for HRV. HRV was detected in 14 NPAs positive and 12 NPAs negative for HBoV. Upon VP4 gene analysis, 5 of these 26 HRV strains were found to belong to HRV-A while 21 belonged to a genetic clade probably representing a previously undetected HRV species, HRV-C, that is phylogenetically distinct from the two known HRV species, HRV-A and HRV-B. The VP4 sequences of these HRV-C strains were closely related to the newly identified HRV strains from the United States and Australia. Febrile wheeze or asthma was the most common presentation (76%) of HRV-C infection, which peaked in fall and winter. Complete genome sequencing of three HRV-C strains revealed that HRV-C represents an additional HRV species, with features distinct from HRV-A and HRV-B. Analysis of VP1 of HRV-C revealed major deletions in regions important for neutralization in other HRVs, which may be signs of a distinct species, while within-clade amino acid variation in potentially antigenic regions may indicate the existence of different serotypes among HRV-C strains. A newly identified HRV species, HRV-C, is circulating worldwide and is an important cause of febrile wheeze and asthmatic exacerbations in children requiring hospitalization.


title: Structural modeling of 2019-novel coronavirus (nCoV) spike protein reveals a proteolytically-sensitive activation loop as a distinguishing feature compared to SARS-CoV and related SARS-like coronaviruses.

text: The 2019 novel coronavirus (2019-nCoV) is currently causing a widespread outbreak centered on Hubei province, China and is a major public health concern. Taxonomically 2019-nCoV is closely related to SARS-CoV and SARS-related bat coronaviruses, and it appears to share a common receptor with SARS-CoV (ACE-2). Here, we perform structural modeling of the 2019-nCoV spike glycoprotein. Our data provide support for the similar receptor utilization between 2019-nCoV and SARS-CoV, despite a relatively low amino acid similarity in the receptor binding module. Compared to SARS-CoV, we identify an extended structural loop containing basic amino acids at the interface of the receptor binding (S1) and fusion (S2) domains, which we predict to be proteolytically-sensitive. We suggest this loop confers fusion activation and entry properties more in line with MERS-CoV and other coronaviruses, and that the presence of this structural loop in 2019-nCoV may affect virus stability and transmission.


title: Conservation medicine and a new agenda for emerging diseases.

text: The last three decades have seen an alarming number of high-profile outbreaks of new viruses and other pathogens, many of them emerging from wildlife. Recent outbreaks of SARS, avian influenza, and others highlight emerging zoonotic diseases as one of the key threats to global health. Similar emerging diseases have been reported in wildlife populations, resulting in mass mortalities, population declines, and even extinctions. In this paper, we highlight three examples of emerging pathogens: Nipah and Hendra virus, which emerged in Malaysia and Australia in the 1990s respectively, with recent outbreaks caused by similar viruses in India in 2000 and Bangladesh in 2004; West Nile virus, which emerged in the New World in 1999; and amphibian chytridiomycosis, which has emerged globally as a threat to amphibian populations and a major cause of amphibian population declines. We discuss a new, conservation medicine approach to emerging diseases that integrates veterinary, medical, ecologic, and other sciences in interdisciplinary teams. These teams investigate the causes of emergence, analyze the underlying drivers, and attempt to define common rules governing emergence for human, wildlife, and plant EIDs. The ultimate goal is a risk analysis that allows us to predict future emergence of known and unknown pathogens.


title: Sequence and immunogenicity of a clinically approved novel measles virus vaccine vector.

text: The measles virus vaccine (MVbv) is a clinically certified and well-tolerated vaccine strain that has been given both parenterally and mucosally. It has been extensively used in children and has proven to be safe and effective in eliciting protective immunity. This specific strain was therefore chosen to generate a measles viral vector. The genome of the commercial MVbv vaccine strain was isolated, sequenced and a plasmid, p(+)MVb, enabling transcription of the viral antigenome and rescue of MVb, was constructed. Phylogenic and phenotypic analysis revealed that MVbv and the rescued MVb constitute another evolutionary branch within the hitherto classified measles vaccines. Plasmid p(+)MVb was modified by insertion of artificial MV-type transcription units (ATUs) for the generation of recombinant viruses (rMVb) expressing additional proteins. Replication characteristics and immunogenicity of rMVb vectors were similar to the parental MVbv and to other vaccine strains. The expression of the additional proteins was stable over 10 serial virus transfers, which corresponds to an amplification greater than 10 ( 20) . The excellent safety record and its efficient application as aerosol may add to the usefulness of the derived vectors.


title: Clinical canine parvovirus type 2C infection in a group of Asian small-clawed otters (Aonyx cinerea).

text: Despite the occurrence of clinical disease in a wide range of carnivore hosts, only vague accounts of clinical canine parvovirus type 2 (CPV-2) in any otter species have been reported in the literature. Over the course of 25 days, nine Asian small-clawed otters (Aonyx cinerea) presented for evaluation of inappetence, lethargy, vomiting, and diarrhea. A diagnosis of canine parvovirus type 2c was made based on electron microscopy, polymerase chain reaction, and DNA sequencing of group fecal samples. Supportive care was provided based on individual clinical assessment and included subcutaneous crystalline fluid therapy, antiemetics, antibiotics, appetite stimulants, and a neuraminidase inhibitor. Five of the nine otters exhibited moderate to severe disease requiring treatment, and one case was fatal despite supportive efforts. In light of this case report, CPV-2 should be recognized as a potential cause of gastrointestinal disease in Asian small-clawed otters.


Host and environment are key factors.


title: Door-to-Needle Time Under 60 Minutes and Picture-to-Puncture Under 90 Minutes: Initiatives and Outcomes in Reducing Time to Recanalization for Cerebral Major Artery Occlusion.

text: The adoption of stent retrievers has significantly improved outcomes of intravenous treatment for acute stroke due to major artery occlusion, and reducing the time to recanalization may achieve further improvements. We reviewed reductions in "door-to-needle time" (DNT) and "picture-to-puncture time" (P2P), as the results of measures to consolidate stroke response capabilities in our hospital, and compared treatment outcomes in acute recanalization patients. We investigated DNT by the route of admission for 96 consecutive patients who received intravenous tissue plasminogen activator between July 2012 and June 2015. We then retrospectively studied 52 patients with acute stroke who underwent endovascular recanalization within 8 h after stroke onset, grouped according to recanalization before (Group I; n = 23) or after (Group II; n = 29) introduction of stent retrievers. Between 2012 and 2015, mean DNT decreased. Significant differences between Groups I and II were only seen in times required, with significantly shorter DNT, picture-to-puncture time, admission to puncture time, and puncture to guiding catheter placement time in Group II. A considerable difference in DNT was seen according to the route of patient admission, and consolidation of hospital stroke response capability successfully reduced the time from admission to recanalization.


title: Hematology and blood chemistry parameters differ in free-ranging maned wolves (Chrysocyon brachyurus) living in the Serra da Canastra National Park versus adjacent farmlands, Brazil.

text: There has been growing interest in the specific impacts of anthropogenic factors on the health of wildlife. This study examined hematology and serum chemistry status of a prominent carnivore, the maned wolf (Chrysocyon brachyurus), living in, on the boundaries to, or on adjacent farmlands to the Serra da Canastra National Park, Brazil. Twenty-eighty wolves were captured, and values were compared 1) between subadults (n=8 animals) and adults (n=20 animals), 2) males (n=12 animals) and females (n=16 animals), and 3) among wolves living inside the park (n=11), near the park border (n=11 animals), and in neighboring farming areas (n=6 animals). Age, gender, and wolf locations influenced (P<0.05) hematology and serum biochemistry values. Specifically, adults had lower (P<0.05) circulating phosphorus than subadults. Males had lower (P<0.05) serum glucose, creatinine phosphokinase, and cholesterol and higher (P<0.05) potassium than females. Erythrocyte count and serum cholinesterase were lower (P<0.05) in wolves living within the park compared with near the park border or on farmlands. Mean corpuscular volume was lower (P<0.05) in wolves living near the park border than those ranging within the park and on farmlands. Aspartate transaminase and chloride were higher (P<0.05) in wolves living inside the park compared with those ranging near the park border. Creatinine phosphokinase was lower (P<0.05) in wolves living on farmland compared with the other two locations. These results clearly reveal a relationship between age and gender on hematology and serum biochemistry values in free-living maned wolves. More importantly, certain traits indicative of health are potentially compromised in wolves living in areas under anthropogenic pressure. These data lay a foundation for examining the influence of farming and local domestic species on disease susceptibility and fitness in the maned wolf.


title: Pro-calcitonin and inflammation in chronic hemodialysis.

text: UNLABELLED Procalcitonin (PCT) has emerged as a marker of infection, a frequent complication in hemodialysis (HD). We analyzed PCT levels in chronic non-acutely infected HD subjects, assessed its correlation with inflammatory and nutritional markers and propose a PCT reference value for non-infected HD patients. In an observational cross-sectional study, 48 chronic HD patients and 36 controls were analyzed. VARIABLES age, gender, time on HD; diabetes; vascular access, PCT, C-reactive protein (CRP), albumin, malnutrition inflammatory score (MIS), hematocrit, leukocyte count, and body mass index (BMI). Subsequently, control (G1, n = 36, 43%) vs. non-infected patients (G2, n = 48, 57%) groups were compared. In control subjects (G1), age: 54.3 ± 13.7 years, range (r): 30-81; males: 19 (53%); median PCT 0.034 ng/ml (r: 0.02-0.08); median CRP 0.80 mg/ dl (r: 0.36-3.9); p95 PCT level: 0.063 ng/ml. In G2, age: 60.2 ± 15.2 years; males 32 (67%), time on HD: 27.0 ± 24.4; diabetics: 19 (32%); median PCT: 0.26 ng/ml (r: 0.09-0.82); CRP: 1.1 mg/dl (r: 0.5-6.2); p95 PCT level: 0.8 ng/ml. In control subjects, PCT and CRP were significantly lower than in G2: PCT: 0.034 vs. 0.26 ng/ml, p = 0.0001; CRP: 0.8 vs. 1.1 mg/dl, p = 0.0004. PCT-CRP correlation in G2: p = 0.287, p = 0.048. PCT and CRP concentrations are elevated in chronic non-acutely infected HD subjects, independently of infection, diabetes and vascular access. A p95 PCT level of 0.8 ng/ml may be considered as the upper normal reference value in non-acutely infected HD subjects. The PCT cut-off level in HD is yet to be determined in HD.


title: Individualizing the use of negative pressure wound therapy for optimal wound healing: a focused review of the literature.

text: Currently available research suggests that negative pressure wound therapy (NPWT) creates a moist wound healing environment, drains exudate, reduces tissue edema, contracts the wound edges, mechanically stimulates the wound bed, and influences blood perfusion at the wound edge, which may lead to angiogenesis and the formation of granulation tissue. Although no clear evidence is available that NPWT accelerates wound healing compared to other interventions or that one form of NPWT is better than another, preclinical research suggests that the most commonly used dressings, level of negative pressure, and application mode (continuous, intermittent, or variable) may not be optimal for all patients. To summarize available literature related to these NPWT choices, pertinent literature published between 2005 and 2010 was reviewed. Preclinical study results suggest that the maximal biological effect of NPWT at the wound edge often can be achieved at -80 mm Hg and that foam dressings may be advantageous for large defect wounds, whereas gauze dressings may be more suitable for smaller wounds or when scar formation or pain is a concern. Preclinical research results also suggest that intermittent or variable pressure application has a better effect on granulation tissue formation than continuous application. The variable pressure mode maintains a negative pressure environment at lower pressure settings without dramatic fluctuations inherent to intermittent (on-and-off) pressure. Prospective, controlled clinical studies are needed to compare NPWT to other advanced wound care protocols of care and to ascertain the effect of various NPWT methods and regimens on outcomes of care.


title: Biomarker discovery in infectious diseases using SELDI.

text: Surface enhanced laser desorption ionization-time of flight is a mass spectrometric-based method that requires a minimal amount of sample for analysis and can be used for high-throughput screening. It has been used to discover serum or tissue protein signatures and biomarkers for infectious diseases in the fields of virology (hepatitis B and C viruses, severe acute respiratory syndrome, HIV-1, human T-cell leukemia virus-1 and BK virus), parasitology (trypanosomiasis) and bacteriology (intra-amniotic inflammation, tuberculosis and bacterial endocarditis). The protein signatures, or biomarkers, can be used to diagnose infection, predict disease states and to inform on disease processes. Careful attention to experimental design, sample handling and storage, and the use of appropriate internal controls is crucial to success.


title: Molecular cloning and characterization of lysosomal sialic acid O-acetylesterase.

text: O-Acetylation and de-O-acetylation of sialic acids have been implicated in the regulation of a variety of biological phenomena, including endogenous lectin recognition, tumor antigenicity, virus binding, and complement activation. Applying a strategy designed to identify genes preferentially expressed in active sites of embryonic hematopoiesis, we isolated a novel cDNA from the pluripotent hematopoietic cell line FDCPmixA4 whose open reading frame contained sequences homologous to peptide fragments of a lysosomal sialic acid O-acetylesterase (Lse) previously purified from rat liver, but with no evident similarity to endoplasmic reticulum-derived acetylesterases. The expressed Lse protein exhibits sialic-acid O-acetylesterase activity that is not attributable to a typical serine esterase active site. lse expression is spatially and temporally restricted during embryogenesis, and its mRNA levels correlate with differences in O-acetylesterase activity described in adult tissues and blood cell types. Using interspecific backcross analysis, we further mapped the lse gene to the central region of mouse chromosome 9. This constitutes the first report on the molecular cloning of a sialic acid-specific O-acetylesterase in vertebrates and suggests novel roles for the 9-O-acetyl modification of sialic acids during the development and differentiation of mammalian organisms.


Respiratory precautions for MERS-CoV: acceptable risk-benefit determination.


title: Flammability of Respirators and other Head and Facial Personal Protective Equipment.

text: Background Personal protective equipment (PPE) is worn by workers in surgical settings to protect them and patients. Food and Drug Administration (FDA) clears some PPE (e.g., surgical masks (SM)) as class II medical devices, and regulates some (e.g. surgical head cover) as class I exempt devices. For respiratory protection, National Institute for Occupational Safety and Health (NIOSH)-approved N95 filtering facepiece respirators (FFRs), and powered air-purifying respirators (PAPRs) are used. One type of PPE, "surgical N95 respirators", is a NIOSH-approved FFR that is also cleared by the FDA for use in medical settings. The surgical environment poses unique risks such as the potential for surgical fires. As part of its substantial equivalence determination process, FDA requests testing of flammability and other parameters for SM and surgical N95 respirators. A lack of data regarding flammability of PPE used in healthcare exists. We hypothesize that commonly used PPE, regardless of whether regulated and/or cleared by FDA or not, will pass an industry standard such as the 16 CFR 1610 flammability test. Methods Eleven N95 FFR models, eight surgical N95 respirator models, seven SM models, five surgical head cover models, and five PAPR hood models were evaluated for flammability with a 45 degree flammability tester using the 16 CFR 1610 method. Three common fabrics were included for comparison. Results All of the PPE samples regulated/and or cleared by FDA or not, passed the flammability test at class 1 (normal flammability), meaning they are less likely to burn. Only one of the three common fabrics, a cotton fabric at the lowest basis weight, was class 3 (high flammability). Conclusions The results obtained in the study suggest that NIOSH-approved N95 FFRs would likely pass the 16 CFR 1610 flammability standard. Moreover, results suggest that NIOSH is capable of undertaking flammability testing using the 16 CFR 1610 standard as the flammability results NIOSH obtained for N95 FFRs were comparable to the results obtained by a third party independent laboratory.


title: Safety and effectiveness of mechanical versus hand suturing of intestinal anastomoses in an animal model of peritonitis.

text: Mechanical stapling for colorectal anastomosis is popular, but the safety of its use for anastomosis in peritonitis is unclear. We evaluated the safety and effectiveness of mechanically stapled vs. hand-sutured anastomosis by comparing wound healing in an animal model of bacterial peritonitis. Male Sprague-Dawley (n=48) rats underwent cecal ligation and puncture. After 24 h, rats were divided into two groups: the stapler group (cecal resection with mechanical stapler, n=24) and the hand-sutured group (cecal resection and stump closure with surgical absorbable suture, n=24). Anastomotic segments were excised and as indicators of wound healing, anastomotic bursting pressure (ABP) and tissue hydroxyproline concentration were determined over time. After harvesting, anastomotic segments were analyzed by quantitative real-time polymerase chain reaction (PCR) to determine relative expression of transforming growth factor-β(1) (TGF-β(1)) and vascular endothelial growth factor (VEGF) normalized to that of a constitutive gene. The operative time was significantly shorter in the stapler vs. the hand-sutured group. Both groups showed progressive increases in ABP over the postoperative period. ABP was significantly higher in the stapler vs. the hand-sutured group on postoperative days (PODs) 0 and 3. Tissue hydroxyproline concentration increased from POD 7 in both groups, but between-group difference was not significant. Both groups showed progressive increases in TGF-β(1) and VEGF expression during the 7-day postoperative period. On POD 5, TGF-β(1) gene expression was higher in the stapler vs. the hand-sutured group. VEGF gene expression was identical in both groups. In conclusion, anastomosis by stapler is safer and more effective than that by hand suturing in bacterial peritonitis, since it requires less operating time and creates stronger anastomoses in the early postoperative period.


title: Patient data, early SARS epidemic, Taiwan.

text: Of the first 10 patients in the epidemic of severe acute respiratory syndrome (SARS) in Taiwan, 4 were closely associated with a SARS patient in an airplane. Loose stools or diarrhea, hemophagocytosis syndrome, and high serum levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-a associated with lung lesions were found in all 10 patients.


title: Public health department training of emergency medical technicians for bioterrorism and public health emergencies: results of a national assessment.

text: HYPOTHESIS The public health system has a specialized body of knowledge and expertise in bioterrorism and public health emergency management that can assist in the development and delivery of continuing medical education programs to meet the needs of emergency medical service providers. METHODS A nationally representative sample of the basic and paramedic emergency medical service providers in the United States was surveyed to assess whether they had received training in weapons of mass destruction, bioterrorism, chemical terrorism, radiological terrorism, and/or public health emergencies, and how the training was provided. RESULTS Local health departments provided little in the way of training in biologic, chemical, or radiological terrorism to responders (7.4%-14.9%). State health departments provided even less training (6.3%-17.3%) on all topics to emergency medical services providers. Training that was provided by the health department in bioterrorism and public health emergency response was associated with responder comfort in responding to a bioterrorism event (OR = 2.74, 95% CI = 2.68, 2.81). CONCLUSIONS Local and state public health agencies should work with the emergency medical services systems to develop and deliver training with an all-hazards approach to disasters and other public health emergencies.


Cholangiohepatitis associated with adenovirus-like particles in a pygmy goat kid.


title: Folding of the mouse hepatitis virus spike protein and its association with the membrane protein.

text: Coronaviruses are assembled by budding into pre-Golgi membranes. Using different approaches we have demonstrated that the spike (S) protein and the membrane (M) protein of mouse hepatitis virus (MHV) associate to form large complexes. Newly synthesized M was found in these complexes almost immediately after its synthesis, whereas the S protein started to appear in heterocomplexes after 10-20 min. This is consistent with the slow rate of folding of S and with the observation that folding of S preceeds its association with M. While the folding of S involves the formation of multiple disulfide bonds, folding of M is disulfide-independent. This contrast was reflected by the differential sensitivity of the two proteins to reduction with dithiothreitol (DTT). Addition of DTT to the culture medium of MHV-infected cells drastically impaired the folding of S, but not of M. Consequently, the S protein was unable to interact with M. Under these conditions, S stayed in the ER while M was transported efficiently beyond the site of budding to the Golgi complex. We conclude that the association of S with M is an essential step in the formation of the viral envelope and in the accumulation of both proteins at the site of virus assembly.


title: Mutagenesis of the genome of mouse hepatitis virus by targeted RNA recombination.

text: Our laboratory has described a method for introducing site-specific mutations into the genome of the coronavirus mouse hepatitis virus (MHV) by RNA recombination between cotransfected genomic RNA and a synthetic subgenomic mRNA. By using a thermolabile N protein mutant of MHV as the recipient virus and synthetic RNA7 (the mRNA for the nucleocapsid protein N) as the donor, engineered recombinant viruses were selected as heat-stable progeny resulting from cotransfection. We have recently reported an optimization of the efficiency of targeted recombination in this process by using a synthetic defective interfering (DI) RNA in place of RNA7. The frequency of recombination is sufficiently high that recombinants can often be directly identified without employing a thermal selection. We present here a progress report on our use of this system to map MHV mutants and to construct N gene mutants which include (1) a mutant in which the internal open reading frame within the N gene (the I gene) has been disrupted, and (2) a series of recombinants in which portions of the MHV N gene have been replaced by the homologous regions from the N gene of bovine coronavirus. We also report on some mutants we have not been able to construct.


title: Canine adenovirus type 1 in a fennec fox (Vulpes zerda).

text: A 10-mo-old female fennec fox (Vulpes zerda) with drooling suddenly died and was examined postmortem. Histologic examination of different tissue samples was performed. Vacuolar degeneration and diffuse fatty change were observed in the liver. Several diagnostic methods were used to screen for canine parvovirus, canine distemper virus, canine influenza virus, canine coronavirus, canine parainfluenza virus, and canine adenovirus (CAdV). Only CAdV type 1 (CAdV-1) was detected in several organs (liver, lung, brain, kidney, spleen, and heart), and other viruses were not found. CAdV-1 was confirmed by virus isolation and nucleotide sequencing.


title: Urolithiasis in rats consuming a dl bitartrate form of choline in a purified diet.

text: Urolithiasis appeared in rats maintained to study the effects of nutrients and methylmercury on development and aging. After a year, the mortality rate was approximately 10%, and by 2 years, it had increased to nearly 30%. Clinical signs and urinary tract pathology were examined as a function of diet, duration on diet, gender, methylmercury exposure, genetics, and other potential risk factors by using survival analyses and qualitative comparisons. Urolithiasis in female rats appeared 15 weeks after beginning a purified diet and after 5 weeks for male rats. After 97 weeks, the mortality rate of female rats was 22% and for male rats was 64%. Lifetime urolithiasis-associated mortality was about 2% in a group of rats that consumed the contaminated diet for < 30 weeks. No urolithiasis occurred in siblings or cohorts of the rats described here that were maintained on a standard rodent chow containing choline chloride. Urolithiasis was traced to racemic, rather than levo-, bitartaric acid in some purified diets shipped in 2001 and 2002. It is unknown when the impurity first appeared in the diet, so estimates of exposure duration are upper limits. Chronic methylmercury exposure increased vulnerability. Some families (dam + offspring) had multiple cases of urolithiasis, but probability models constructed to evaluate familial clustering revealed no evidence for a genetic predisposition to urolithiasis apart from gender. Removing racemic tartaric acid did not decrease mortality once rats had been on the diet for 20 to 30 weeks, but it helped when exposure duration was shorter.


title: The associations of viral and mycoplasmal antibody titers with respiratory disease and weight gain in feedlot calves.

text: Blood samples from 32 groups of calves (n = 700) were taken on arrival and after 28-35 days at the feedlot. Eleven groups were housed in feedlots in Ontario, and 21 groups in feedlots in Alberta. Serum antibody titers to bovine viral diarrhea virus (BVDV), bovine respiratory syncytial virus (BRSV), parainfluenza virus type 3 (PIV-3), infectious bovine rhinotracheitis virus (IBRV), Mycoplasma dispar and M. bovis, plus data on bovine corona virus (BCV) from a previous study were investigated for their association with the risk of bovine respiratory disease (BRD), and with 28-day weight change, both before and after controlling for titers to Pasteurella haemolytica and Haemophilus somnus. Exposure to IBRV and M. bovis was infrequent, and although exposure to PIV-3 was more common, none of these agents had important associations with BRD. Higher titers to BVDV, BRSV, and BCV on arrival were associated with reduced risks of BRD and increased weight gains. However, there was some variation in these relationships and higher arrival titers to BVDV and BRSV in a subset of the calves were associated with increased risks of BRD. Titer increases to BVDV were associated with a higher risk of BRD and lower weight gains. Titer increases to BRSV were not usually associated with the occurrence of BRD, but titer increases to BRSV in a subset of calves that were vaccinated against BRSV, on arrival, were associated with an elevated risk of BRD. Of all the agents studied, BVDV had the most consistent associations with elevated risk of BRD and lower weight gains. Higher BRSV arrival titers were related to lower risk of BRD and higher weight gains; in some instances titer increases to BRSV were associated with higher BRD risk. Higher titers to BCV on arrival were related to reduced risks of BRD. Practical ways of adequately preventing the negative effects of these agents are still needed.


Childhood obstructive sleep apnoea.


title: [Purification of coronaviruses from contaminated cattle feces].

text: A scheme was worked out for the obtaining of purified Corona viruses from fecal extracts, which include: centrifugation in an discontinued gradient of sucrose 20--45 per cent, gel filtration through a Sephadex 4B column, and centrifugation in a linear gradient of sucrose 10--50 per cent. By means of such purification the virions remain morphologically intact in sucrose density of 1.11--1.14 gr/cm3, containing no ballast proteins. As many as five polypeptides are demonstrated in polyacrylamide electrophoresis.


title: Survey of the year 2006 commercial optical biosensor literature.

text: We identified 1219 articles published in 2006 that described work performed using commercial optical biosensor platforms. It is interesting to witness how the biosensor market is maturing with an increased number of instrument manufacturers offering a wider variety of platforms. However, it is clear from a review of the results presented that the advances in technology are outpacing the skill level of the average biosensor user. While we can track a gradual improvement in the quality of the published work, we clearly have a long way to go before we capitalize on the full potential of biosensor technology. To illustrate what is right with the biosensor literature, we highlight the work of 10 groups who have their eye on the ball. To help out the rest of us who have the lights on but nobody home, we use the literature to address common myths about biosensor technology.


title: Infectious bronchitis virus subpopulations in vaccinated chickens after challenge.

text: Infectious bronchitis coronavirus (IBV) shows extensive genotypic and phenotypic variability. The evolutionary process involves generation of genetic diversity by mutations and recombination followed by replication of those phenotypes favored by selection. In the current study, we examined changes occurring in a wild Arkansas (Ark) challenge strain in chickens that were vaccinated either ocularly with commercially available attenuated ArkDPI-derived vaccines or in ovo with a replication-defective recombinant adenovirus expressing a codon-optimized IBV Ark S1 gene (AdArkIBV.S1(ck)). Commercial IBV Ark vaccines A, B, and C provided slightly differing levels of protection against homologous challenge. Most importantly for the current study, chickens vaccinated with the different vaccines displayed significant differences in specific B-lymphocyte responses in the Harderian gland (i.e., the challenge virus encountered differing immune selective pressure during invasion among host groups). Based on S1 sequences, five predominant populations were found in different individual vaccinated/challenged chickens. Chickens with the strongest immune response (vaccine A) were able to successfully impede replication of the challenge virus in most chickens, and only the population predominant in the challenge strain was detected in a few IBV-positive birds. In contrast, in chickens showing less than optimal specific immune responses (vaccines B and C) IBV was detected in most chickens, and populations different from the predominant one in the challenge strain were selected and became predominant. These results provide scientific evidence for the assumption that poor vaccination contributes to the emergence of new IBV strains via mutation and/or selection. In ovo vaccination with a low dose of AdArkIBV.S1(ck) resulted in a mild increase of systemic antibody and reduced viral shedding but no protection against IBV signs and lesions. Under these conditions we detected only virus populations identical to the challenge virus. Possible explanations are discussed. From a broad perspective, these results indicate that selection is an important force driving IBV evolution.


title: Coronavirus neurovirulence correlates with the ability of the virus to induce proinflammatory cytokine signals from astrocytes and microglia.

text: The molecular and cellular basis of coronavirus neurovirulence is poorly understood. Since neurovirulence may be determined at the early stages of infection of the central nervous system (CNS), we hypothesize that it may depend on the ability of the virus to induce proinflammatory signals from brain cells for the recruitment of blood-derived inflammatory cells. To test this hypothesis, we studied the interaction between coronaviruses (mouse hepatitis virus) of different neurovirulences with primary cell cultures of brain immune cells (astrocytes and microglia) and mouse tissues. We found that the level of neurovirulence of the virus correlates with its differential ability to induce proinflammatory cytokines (interleukin 12 [IL-12] p40, tumor necrosis factor alpha, IL-6, IL-15, and IL-1beta) in astrocytes and microglia and in mouse brains and spinal cords. These findings suggest that coronavirus neurovirulence may depend on a novel discriminatory ability of astrocytes and microglia to induce a proinflammatory response in the CNS.


Characterization of low-virulent mouse coronavirus isolated from faeces in a mouse colony.


title: Effect of time and exercise on the clearance rate of (201)Tl in normal and ischemic myocardium.

text: PURPOSE Simultaneous dual isotope (SDI) acquisition of (201)Tl rest/(99m)Tc-sestamibi stress-myocardial perfusion single photon emission computed tomography is a desirable new procedure in nuclear cardiology. In this protocol (201)Tl is injected at rest but imaging is performed not earlier than after exercise. Therefore, one must be convinced that throughout exercise (201)Tl remains distributed in an identical pattern as at rest. Before SDI can be applied clinically, (201)Tl rest MPS before and after exercise test needs to be compared for equality. The aim of this study was to assess the effect of time and exercise on the clearance of preinjected (201)Tl in normal and ischemic myocardium. METHODS In 122 patients rest (201)Tl and delayed (n =20) or poststress (n= 102) (201)Tl imaging was performed. Quantitative analysis of mean counts-per-pixel was performed for each segment in a 17-segment model. Differences between rest and delayed or poststress (201)Tl MPS were calculated. Patients with a poststress (201)Tl image were divided into normal (N= 66) and ischemic (N= 36) groups. Visual analysis was performed by two independent observers scoring the 17 segments on a scale of 0-4. RESULTS The overall difference between rest (201)Tl and poststress (201)Tl MPS was - 15.4%. Normal and ischemic patients showed 16.2 and 14.0% (P =0.17) washout, respectively. Visual assessment by two independent observers revealed no regional differences between rest (201)Tl and delayed or poststress (201)Tl MPS. CONCLUSION (201)Tl poststress MPS shows significant washout of thallium. This washout is not segmental, but global over the myocardium. No significant differences are found between normal and ischemic myocardium. The poststress (201)Tl MPS is a reliable reflection of rest perfusion. SDI acquisition of (201)Tl rest/(99m)Tc-sestamibi stress-MPS is clinically applicable.


Use of drotrecogin alfa (activated) in a severe acute respiratory syndrome patient with severe sepsis.


title: Randomized controlled trial of foot reflexology for patients with symptomatic idiopathic detrusor overactivity.

text: The aim of this study was to examine whether foot reflexology has beneficial effects on patients with idiopathic detrusor overactivity. One hundred and nine women with symptomatic idiopathic detrusor overactivity were randomized into either foot reflexology treatment group or nonspecific foot massage control group. The primary outcome measure was the change in the diurnal micturition frequency. There was significant change in the number of daytime frequency in the reflexology group when compared with the massage group (-1.90 vs -0.55, p = 0.029). There was also a decrease in the 24-h micturition frequency in both groups, but the change was not statistically significant (-2.80 vs -1.04 p = 0.055). In the reflexology group, more patients believed to have received "true" reflexology (88.9 vs 67.4%, p = 0.012). This reflects the difficulty of blinding in trials of reflexology. Larger scale studies with a better-designed control group and an improved blinding are required to examine if reflexology is effective in improving patients' overall outcome.


title: SARS: UK public health response--past, present and future.

text: The emergence of severe acute respiratory syndrome (SARS) in China, the occurrence of epidemics of SARS in China and a number of Southeast Asian countries, and its spread to countries elsewhere, have presented major challenges to public health systems throughout the world. Although very few true cases of SARS were detected in the United Kingdom, the public health response to the threat of SARS was considerable. The main components of this response were the early detection, isolation and reporting of cases, and the provision of comprehensive information to health professionals, cases, their contacts and the public. The development of the response to SARS raised a number of more general issues relevant to future infectious epidemic threats. Although the World Health Organisation has now declared SARS 'contained', the possibility of re-emergence is ever present. All countries will need to be vigilant and plan their response to the possibility of a renewed SARS epidemic.


title: Spike glycoprotein-mediated fusion in biliary glycoprotein-independent cell-associated spread of mouse hepatitis virus infection.

text: The mouse hepatitis virus (MHV) spike glycoprotein mediates attachment of the virus to the MHV receptor, the murine biliary glycoprotein (BGP) carcinoembryonic antigen. Monoclonal antibody CC1 directed against BGP specifically inhibited infection of DBT, Sac-, GT1-7, and OBL21 cells by wild-type MHV-4 and the neuron-adapted variant OBLV60. Binding to this receptor was necessary to establish infection by cell-free MHV; however, the presence of BGP was not required for infection by cell-associated virus. Cell-associated infectious induced syncytium formation on Vero and BHK cells, which lack murine BGP; this activity was not inhibited by monoclonal antibody CC1. Antibody CC1 also did not prevent syncytium formation on DBT cells, which bear BGP. In infectious center assays, the MHV-4 variant OBLV60, which exhibits acid-dependent fusion, spread to cells lacking BGP only when exposed to acidic media. Therefore, spike-mediated fusion was required for BGP-independent spread of MHV infection. Furthermore, BGP-independent, cell-associated spread of MHV-4 was prevented by monoclonal antibodies 5A13.5 and 5B19.2 directed against the spike glycoprotein, but not by other neutralizing and nonneutralizing anti-spike antibodies. Expression of spike glycoprotein by recombinant vaccinia virus resulted in fusion of BGP-negative cells; monoclonal antibodies 5A13.5 and 5B19.2 strongly inhibited spike-mediated fusion in this assay.


The biology and pathogenesis of coronaviruses.


RNA virus replication, transcription and recombination.


title: The use of the Veress needle to drain mammary periprosthetic fluid.

text: BACKGROUND The most common early complications associated with mammary implants are seroma and haematoma formation, and acute infection. The aspiration of fluid around the prosthesis can help to diagnose and correctly treat these complications. The use of a needle with a sharp tip can damage the implant. We present our experience in removing periprosthetic fluid using the atraumatic Veress needle. PATIENTS AND METHODS Twelve patients with breast implants presented with a progressive monolateral breast enlargement. Ultrasound examination revealed the collection of fluid around the implants. Ultrasound-guided percutaneous needle aspiration of the fluid was performed using the Veress needle. RESULTS No complications were reported. None of the implants was found to be damaged at the ultrasound assessment. CONCLUSION Our proposed use of the Veress needle is similar to the use for which it was designed. Indeed, we used the device to remove some fluid from a real cavity. This procedure proved to be effective and safe.


title: Detection of U.S., Lelystad, and European-like porcine reproductive and respiratory syndrome viruses and relative quantitation in boar semen and serum samples by real-time PCR.

text: Transmission of porcine reproductive and respiratory syndrome virus (PRRSV) via boar semen has been documented. Since semen is widely disseminated for artificial insemination and the virus can cause significant health and economic consequences, it is essential to have well-validated, rapid diagnostic techniques to detect and quantitate the virus for diagnostic and research purposes. Previously, boar semen was tested by a nested PCR (nPCR) assay which was compared to the "gold standard" swine bioassay. A correlation of 94% was observed, indicating that, most of the time, PCR detected infectious virus. Subsequently, a real-time PCR targeting the 3' untranslated region of the PRRSV genome was compared with nPCR by testing 413 serum and semen samples from PRRSV-inoculated and control boars. There was 95% agreement between the results of the two tests, with the majority of samples with discordant results containing virus at the lower range of detection by the assays. The virus in all samples was quantitated by using a standard curve obtained by serial dilution of an in vitro transcript. By using the in vitro transcript, the lower limit of sensitivity was observed to be approximately 33 copies/ml. Reactivity with a panel of more than 100 PRRSV isolates from various geographical regions in the United States was also documented. No reactivity with nine nonrelated swine viruses was noted. A real-time PCR was also developed for the detection of the European Lelystad virus and the European-like PRRSV now found in the United States. In six of six PRRSV-inoculated boars, peak levels of viremia occurred at 5 days postinoculation (DPI) and were most consistently detectable throughout 22 DPI. In five of six boars, PRRSV was shed in semen for 0 to 2 days during the first 10 DPI; however, one of six boars shed the virus in semen through 32 DPI. Therefore, in general, the concentration and duration of PRRSV shedding in semen did not correlate with the quantity or duration of virus in serum. These differences warrant further studies into the factors that prevent viral replication in the reproductive tract.


title: Team Epi-Aid: graduate student assistance with urgent public health response.

text: Team Epi-Aid provides graduate students with practical public health experience through participation in outbreak investigations and other applied projects with state and local health departments in North Carolina. It is an initiative of the North Carolina Center for Public Health Preparedness in the North Carolina Institute for Public Health at the University of North Carolina School of Public Health. The program allows state and local health departments access to volunteers and technical expertise from the university when they need assistance. It requires close collaboration with state and county health departments. Team Epi-Aid provides the opportunity for integrated learning with students and faculty within the departments of the School of Public Health, and through recent expansion, within the schools of Medicine and Pharmacy. Orientations are conducted each semester and formal training is provided as needed. Team Epi-Aid has been popular, with 58 active student participants contributing 1,465 hours of service during the initiative's first 21 months.


title: SARS in Singapore--key lessons from an epidemic.

text: The rapid containment of the Singapore severe acute respiratory syndrome (SARS) outbreak in 2003 involved the introduction of several stringent control measures. These measures had a profound impact on the healthcare system and community, and were associated with significant disruptions to normal life, business and social intercourse. An assessment of the relative effectiveness of the various control measures is critical in preparing for future outbreaks of a similar nature. The very "wide-net" surveillance, isolation and quarantine policy adopted was effective in ensuring progressively earlier isolation of probable SARS cases. However, it resulted in nearly 8000 contacts being put on home quarantine and 4300 on telephone surveillance, with 58 individuals eventually being diagnosed with probable SARS. A key challenge is to develop very rapid and highly sensitive tests for SARS infection, which would substantially reduce the numbers of individuals that need to be quarantined without decreasing the effectiveness of the measure. Daily temperature monitoring of all healthcare workers (HCWs) in hospitals was useful for early identification of HCWs with SARS. However, daily temperature screening of children in schools failed to pick up any SARS cases. Similarly, temperature screening at the airport and other points of entry did not yield any SARS cases. Nevertheless, the latter 2 measures probably helped to reassure the public that schools and the community were safe during the SARS outbreak. Strong political leadership and effective command, control and coordination of responses were critical factors for the containment of the outbreak.


title: Conformational changes in the spike glycoprotein of murine coronavirus are induced at 37 degrees C either by soluble murine CEACAM1 receptors or by pH 8.

text: The spike glycoprotein (S) of the murine coronavirus mouse hepatitis virus (MHV) binds to viral murine CEACAM receptor glycoproteins and causes membrane fusion. On virions, the 180-kDa S glycoprotein of the MHV-A59 strain can be cleaved by trypsin to form the 90-kDa N-terminal receptor-binding subunit (S1) and the 90-kDa membrane-anchored fusion subunit (S2). Incubation of virions with purified, soluble CEACAM1a receptor proteins at 37 degrees C and pH 6.5 neutralizes virus infectivity (B. D. Zelus, D. R. Wessner, R. K. Williams, M. N. Pensiero, F. T. Phibbs, M. deSouza, G. S. Dveksler, and K. V. Holmes, J. Virol. 72:7237-7244, 1998). We used liposome flotation and protease sensitivity assays to investigate the mechanism of receptor-induced, temperature-dependent virus neutralization. After incubation with soluble receptor at 37 degrees C and pH 6.5, virions became hydrophobic and bound to liposomes. Receptor binding induced a profound, apparently irreversible conformational change in S on the viral envelope that allowed S2, but not S1, to be degraded by trypsin at 4 degrees C. Various murine CEACAM proteins triggered conformational changes in S on recombinant MHV strains expressing S glycoproteins of MHV-A59 or MHV-4 (MHV-JHM) with the same specificities as seen for virus neutralization and virus-receptor activities. Increased hydrophobicity of virions and conformational change in S2 of MHV-A59 could also be induced by incubating virions at pH 8 and 37 degrees C, without soluble receptor. Surprisingly, the S protein of recombinant MHV-A59 virions with a mutation, H716D, that precluded cleavage between S1 and S2 could also be triggered to undergo a conformational change at 37 degrees C by soluble receptor at neutral pH or by pH 8 alone. A novel 120-kDa subunit was formed following incubation of the receptor-triggered S(A59)H716D virions with trypsin at 4 degrees C. The data show that unlike class 1 fusion glycoproteins of other enveloped viruses, the murine coronavirus S protein can be triggered to a membrane-binding conformation at 37 degrees C either by soluble receptor at neutral pH or by alkaline pH alone, without requiring previous activation by cleavage between S1 and S2.


title: Coronavirus isolation from nasal swab samples in cattle with signs of respiratory tract disease after shipping.

text: OBJECTIVE To monitor the prevailing viral respiratory tract infections in cattle after transportation to feedlots. ANIMALS 100 cattle with signs of respiratory tract disease on arrival at 2 feedlots. PROCEDURES Nasal swab samples were obtained from each animal and were used for inoculation of defined cell culture systems that detected bovine viruses known to cause respiratory tract infections, as well as viruses previously not recognized as respiratory pathogens for cattle. RESULTS Bovine respiratory coronaviruses were isolated from 38 of the 100 cattle, including 6 of 50 cattle from California, 22 of 31 cattle from Oklahoma, 6 of 11 cattle from Texas, and 4 of 8 cattle of unknown origin. Parainfluenza 3 viruses also were isolated from 5 California cattle, but other bovine viruses were not detected. CLINICAL IMPLICATIONS The high rate of coronavirus isolations from feedlot cattle with signs of respiratory tract disease implied wide distribution and high susceptibility among cattle to this infection, which had not been detected by use of viral isolation systems in previous etiologic evaluations of feedlot cattle affected with bovine respiratory disease complex.


title: Neutrophil/lymphocyte and C-reactive protein/mean platelet volume ratios in differentiating between viral and bacterial pneumonias and diagnosing early complications in children.

text: OBJECTIVE To test the usability of neutrophil/lymphocyte (N/L) and C-reactive protein/mean platelet volume (CRP/MPV) ratios for the differential diagnosis of bacterial versus viral pneumonia, and the early diagnosis of complications related to pneumonia. METHODS This retrospective study was conducted on 31 patients diagnosed with bacterial pneumonia and 21 patients diagnosed with viral pneumonia from January 2011 to December 2012 in the Department of Pediatrics, Faculty of Medicine, Abant Izzet Baysal University, Bolu, Turkey. We investigated the clinical characteristics, radiological, and laboratory findings of patients from their medical records. RESULTS The female/male ratio of patients with bacterial was 1.0/1.8, and with viral pneumonias was 1.0/2.0. The mean patient age was 59+/-51 months. There was a statistically significant difference in the neutrophil/lymphocyte ratio (2.7 versus 0.6, p<0.001) and CRP/MPV ratio (11.0 versus 9.3, p<0.001) in the cases with bacterial pneumonia versus those who had viral pneumonia. Nine of the patients were identified as having complications. There was a statistically significant difference in the N/L ratio (3.5 versus 1.2, p=0.01) and CRP/MPV ratio (11.1 versus 3.9, p=0.001) in the cases that developed complications compared with those that did not. When the neutrophil/lymphocyte and CRP/MPV ratios were used jointly, the diagnosis of bacterial pneumonia could be correctly estimated in 28 (90.3%) cases (odds ratio [OR]=0.06, 95% confidence interval [CI]: 0.01-0.29, p<0.001) and pneumonia-related complications were predicted in 8 (88.9%) cases (OR=13.5, 95% CI: 1.5-118.1, p=0.005). CONCLUSION It was observed that the combined use of N/L and CRP/MPV ratios might be used in both the differential diagnosis of bacterial versus viral pneumonia, and the prediction of complications.


title: Mapping of medical acronyms and initialisms to Medical Subject Headings (MeSH) across selected systems.

text: INTRODUCTION Given the common use of acronyms and initialisms in the health sciences, searchers may be entering these abbreviated terms rather than full phrases when searching online systems. The purpose of this study is to evaluate how various MEDLINE Medical Subject Headings (MeSH) interfaces map acronyms and initialisms to the MeSH vocabulary. METHODS The interfaces used in this study were: the PubMed MeSH database, the PubMed Automatic Term Mapping feature, the NLM Gateway Term Finder, and Ovid MEDLINE. Acronyms and initialisms were randomly selected from 2 print sources. The test data set included 415 randomly selected acronyms and initialisms whose related meanings were found to be MeSH terms. Each acronym and initialism was entered into each MEDLINE MeSH interface to determine if it mapped to the corresponding MeSH term. Separately, 46 commonly used acronyms and initialisms were tested. RESULTS While performance differed widely, the success rates were low across all interfaces for the randomly selected terms. The common acronyms and initialisms tested at higher success rates across the interfaces, but the differences between the interfaces remained. CONCLUSION Online interfaces do not always map medical acronyms and initialisms to their corresponding MeSH phrases. This may lead to inaccurate results and missed information if acronyms and initialisms are used in search strategies.


title: Detection of the K99 antigen of Escherichia coli in calf faeces by enzyme-linked immunosorbent assay (ELISA).

text: An enzyme-linked immunosorbent assay (ELISA) is presented for the detection of the K99 antigen of Escherichia coli in calf faeces. False-positive reactions were not observed with K99-negative strains and with several viral antigens. Only bovine coronavirus caused slight positive reactions which could be eliminated by a blocking test. As compared with the conventional procedure for the detection of the K99 antigen, ELISA seemed to be a least as sensitive and had the advantage that samples could be stored at --20 degrees C before testing. In addition many samples could be handled at the same time and the results became available quickly. By carrying out the assay as a blocking test, specific antibody against K99 in serum or colostrum could be detected and titrated.


title: Interspecies aminopeptidase-N chimeras reveal species-specific receptor recognition by canine coronavirus, feline infectious peritonitis virus, and transmissible gastroenteritis virus.

text: We report that cells refractory to canine coronavirus (CCV) and feline infectious peritonitis virus (FIPV) became susceptible when transfected with a chimeric aminopeptidase-N (APN) cDNA containing a canine domain between residues 643 and 841. This finding shows that APN recognition by these viruses is species related and associated with this C-terminal domain. The human/canine APN chimera was also able to confer susceptibility to the porcine transmissible gastroenteritis virus (TGEV), whereas its human/porcine homolog failed to confer susceptibility to CCV and FIPV. A good correlation was observed between the capacity of CCV, FIPV, and TGEV to recognize the different interspecies APN chimeras and their ability to infect cells derived from the relevant species. As an exception, TGEV was found to use a human/bovine APN chimera as a receptor although itself unable to replicate in bovine cells.


Restriction endonuclease analysis of feline herpesvirus 1 DNA isolated from wild felids.


title: [Analysis on the cost and its related factors of clinically confirmeds severe acute respiratory syndrome cases in Beijing].

text: OBJECTIVE To explore the medical cost and its related factors of clinically confirmed severe acute respiratory syndrome (SARS) cases in Beijing. METHODS The average medical cost per person/day was calculated on 1272 SARS cases with complete hospitalization data from the Beijing SARS clinical database. Factors associated with medical expenditure were explored by both single factor and multi-factor analyses. RESULTS The medical costs of SARS patients in Beijing were 18 767.50 RMB/case and 632.02 RMB/day/case. Expenses on medication accounted for the highest proportion (0.66%). The increase in expenses on medication was significantly associated to the increase in total expenditure. Analyses by single factor, factor stratification as well as multi-factor analyses showed that: the expenditure increased significantly with the following factors of the patients: prolonged days of hospitalization, severe symptoms and signs, hospitalization at the early stage of disease onset, being medical staff themselves, with baseline diseases, being applied mechanical ventilation, and with medical insurance. CONCLUSIONS The medical expenditure on SARS patients at the hospital was higher than that on patients with common diseases which was associated to the following factors as: days of hospitalization, severity of disease, stage of the disease break-out, patient's occupation, baseline disease status, medical insurance status etc.


title: [Detection of free cancer cells in peritoneal cavity in patients surgically treated for gastric adenocarcinoma].

text: BACKGROUND/AIM Peritoneal metastasis is a leading cause of therapeutic failure after an operative treatment of patients with gastric adenocarcinoma. Free cancer cells might induce or indicate an early peritoneal seeding with a subsequent peritoneal metastasis. The aim of this study was to determine the frequency of the presence of free cancer cells in the peritoneal cavity in the patients surgically treated for gastric adenocarcinoma, and its relation to certain clinical, operative and pathohistological paramethers. METHODS Inside a period from April 2000, and April 2004, the total of 100 patients underwent intraoperative peritoneal lavage for cytological examination. Immediately after the laparotomy, 200 ml physiologic saline, heated to 37 degrees C, was introduced into the abdominal cavity, mannualy dispersed and collected from the region around the gastric tumor and the pouch of Douglas. The nucleated cell layer was smeared on four glass slides for every patient and dyed with May-Grünwald-Giemsa stain. The cytological findings were defined as positive or negative according to the presence of cancer cells. The frequency of positive cytological findings was compared to the location and the diameter of the cancer, pathohistological type of carcinoma, pathohistological stage of the disease, lymph node and the liver and/or peritoneal metastases and the type of surgical procedure. RESULTS Free cancer cells were found in 24 (24%) of the patients, while in 76 (76%) of them cytological findings were negative. A statistically highly significant difference (p < or = 0.001) in the frequency of positive cytological finding was found between the groups of patients with and without cancer invasion of serosa, with cancer diameters > 5 cm and < or = 5 cm, in the stage of disease I, II and III, IV, with macroscopically present and without metastases, with re section and D2 lymphadenectomy and palliative procedure. Free cancer cells were statistically more frequently (p < or = 0.05) detected in the patients with lymph nodes metastases comparing to the patients with out lymph nodes involvement. The results of the univariate analysis showed that the cancer diameter > 5 cm, tumor invasion of serosa, pathohistological stage of the disease III and IV and macroscopically visible metastases were the most important risk factors for the free cancer cells detection. CONCLUSION Peritoneal lavage cytology was shown to be a useful tool for the detection of the group of patients with greatest risk of peritoneal dissemination. The frequency of positive cytological findings was highly associated with the diameter of the tumor and the cancer invasion of serosa. Cytological examination of peritoneal lavage fluid improved the accuracy of staging and selection of patients who might have benefit from neoadjuvant chemotherapy.


title: Detection of respiratory and enteric shedding of bovine coronaviruses in cattle in Northwestern Turkey.

text: Bovine coronavirus (BCoV) is an important cause of diarrhoea in calves, winter dysentery in adult cattle and respiratory tract disease in feedlot cattle. Serum, faecal and nasal swab samples were collected from a total of 96 cattle with clinical signs in 29 barns of 23 villages in Northwestern Turkey. The cattle were subdivided into 3 distinct age groups (0-30 days old, 4-12 months old and 2-7 years old). An indirect antigen-capture ELISA and an antibody-detection ELISA as well as geometric mean BCoV antibody titres were used to detect BoCV shed in the faeces and in the nasal secretions, respectively. Relationships between BCoV shedding and age group, seroconversion and clinical signs in cattle were also analysed. The rate of faecal shedding of BoCV was 37.1% (13/35) in 0-30 days old calves, 25.6% (10/39) in 4-12 months old feedlot cattle and 18.2% (4/22) in 2-7 years old cows. The overall rate of BCoV faecal shedding was 28.1% (27/96) in the cattle examined. Only one animal in the 4-12 months old age group was found to shed BoCV nasally. The analysis showed that there was a significant difference (P < 0.0001) with respect to faecal shedding between the clinical signs and the age groups. BCoV antibody titre in 50% of all cattle was < or =100 as detected by ELISA while 27.1% of the cattle had high titres ranging between 1,600 and 25,600. The seroconversion rate was 7.3% (7/96) in animals shedding BoCV in the faeces and 42.7% (41/96) in cattle negative for faecal shedding as detected by ELISA, and 20.8% of cattle with no seroconversion shed BCoV in the faeces. There was no statistically significant association between seroconversion and nasal or faecal BCoV shedding. These findings confirm the presence of BCoV infections in Turkey. Further studies are needed to isolate BCoV strains in Turkey and to investigate their antigenic and genetic properties.


Plague in Madagascar:overview of the 2014-2015 epidemic season.


title: "Imaging Evaluation of Collaterals in the Brain: Physiology and Clinical Translation"

text: The cerebral collateral circulation is a network of blood vessels designed to preserve cerebral blood flow when primary routes fail. Though recognized for hundreds of years, the beneficial influence of collateral flow has now gained significant attention due to widely available, rapid, and real-time non-invasive imaging techniques. Multimodal CT and MRI based techniques, with angiographic and perfusion assessments, are becoming mainstays in the care of patients with ischemic brain disease. These methods allow for precise delineation of the structural and functional aspects of cerebral blood flow and as such provide valuable information that can inform the diagnosis and treatment of cerebral ischemia, in both the acute and chronic setting.


title: Serologic response of domestic ferrets (Mustela putorius furo) to canine distemper and rabies virus vaccines.

text: Nine unrelated 12-week-old naive domestic ferrets (Mustela putorius furo) were used to evaluate the serologic responses to commercial canine distemper virus (CDV) and rabies virus (RV) vaccines. Five of the ferrets (group 1) were inoculated 3 times at 2-week intervals with a multivalent modified-live virus vaccine of canine cell-line origin, containing CDV and an inactivated RV vaccine. Four of the ferrets (group 2) were inoculated once with the multivalent modified-live virus vaccine containing CDV and were not inoculated with the RV vaccine. Both group-1 and group-2 ferrets seroconverted to the CDV component of the vaccine. Group-1 ferrets also seroconverted after RV vaccination and maintained serum antibody titers to both CDV and RV for at least 7 months. Domestic ferret sera were found to have IgG epitopes similar to sera of domestic dogs and cats. Domestic ferret sera did not contain antibodies to feline coronavirus or FeLV antigens.


SARS in Northern Vietnam.


Difference in response of susceptible mouse strains to a small plaque mutant (JHM-CC) of mouse coronavirus.


title: The SARS coronavirus nucleocapsid protein induces actin reorganization and apoptosis in COS-1 cells in the absence of growth factors.

text: In March 2003, a novel coronavirus was isolated from patients exhibiting atypical pneumonia, and was subsequently proven to be the causative agent of the disease now referred to as SARS (severe acute respiratory syndrome). The complete genome of the SARS-CoV (SARS coronavirus) has since been sequenced. The SARS-CoV nucleocapsid (SARS-CoV N) protein shares little homology with other members of the coronavirus family. In the present paper, we show that SARS-CoV N is capable of inducing apoptosis of COS-1 monkey kidney cells in the absence of growth factors by down-regulating ERK (extracellular-signal-regulated kinase), up-regulating JNK (c-Jun N-terminal kinase) and p38 MAPK (mitogen-activated protein kinase) pathways, and affecting their downstream effectors. SARS-CoV N expression also down-regulated phospho-Akt and Bcl-2 levels, and activated caspases 3 and 7. However, apoptosis was independent of the p53 and Fas signalling pathways. Furthermore, activation of the p38 MAPK pathway was found to induce actin reorganization in cells devoid of growth factors. At the cytoskeletal level, SARS-CoV N down-regulated FAK (focal adhesion kinase) activity and also down-regulated fibronectin expression. This is the first report showing the ability of the N protein of SARS-CoV to induce apoptosis and actin reorganization in mammalian cells under stressed conditions.


Making the next steps the right ones: progress towards the Smokefree Aotearoa 2025 Goal.


title: Evaluation of serum neutralizing antibodies to bovine coronavirus in cows & their calves using Hmlu-1 cells.

text: Between 1992 and 1993, 75 paired serum samples from Holstein dairy cows and their calves were collected from Aomori, Tochigi and Okinawa Prefectures, and the neutralizing antibody titres to bovine coronavirus (BCV) were determined using hamster lung (Hmlu)-1 cells. The anti-BCV antibody positive rate in the maternal serum samples was significantly lower (P < 0.001) in Okinawa (72%) than in Aomori (100%) or Tochigi (100%). The geometric mean tire (GMT) of anti-BCV neutralizing antibody was also significantly lower (P < 0.05) in maternal sera from Okinawa (89) than that of Aomori (229) or Tochigi (264). The anti-BCV neutralizing antibody titres in the sera of calves which had ingested the colostrum, significantly correlated with the antibody concentration of the maternal serum samples (P < 0.05). These results suggest an extensive BCV infection among the dairy cattle in these prefectures, with a varied pattern of distribution between the prefectures. Anti-BCV neutralizing antibody in the sera of newborn calves appeared to be transferred from their dams through colostrum.


title: Electron microscopy of human immunodeficiency virus.

text: Two cell lines of human T lymphocytes (H9 and CEM) chronically infected with isolates of human or simian immunodeficiency viruses were examined by electron microscopy. Scanning electron microscopy of H9 cells showed characteristic morphological changes in the cells after infection with human T cell lymphotropic virus type III (HTLV-IIIB); these included loss of cell microvilli and their replacement by rounded surface protrusions. Virus particles were present over the whole cell surface, but were more numerous between, rather than on, surface protrusions. In contrast, CEM cells infected with three other isolates of the virus showed little change in morphology compared with uninfected cells; they typically had a single large aggregate of virus particles at the posterior end of the cell. Transmission electron microscopy of sections of infected cells showed budding virus in only a small proportion of cells although many had mature virus particles on their surface. The immature particles released by budding had a ring-like morphology in contrast to the mature virus with its characteristic elongated nucleoid. Surface spikes were rarely seen on sectioned virus particles, but were more obvious on the simian immunodeficiency virus than on human isolates. Negative staining of purified virus preparations showed that the peripheral dense material of immature particles had a striated structure. Surface spikes were not seen by negative staining on either immature or mature virus particles. A third type of smaller particle with surface spikes was found in all negatively stained preparations.


Bird flu: pandemic flu preparation: an unheeded lesson from SARS.


title: A serologic assessment of exposure to viral pathogens and Leptospira in an urban raccoon (Procyon lotor) population inhabiting a large zoological park.

text: In urban environments, raccoons (Procyon lotor) may act as reservoirs for an array of pathogenic organisms, presenting spillover risks for human, domestic animal, and captive (zoo) animal populations. Over 5 yr, 159 raccoons from a high-density raccoon population in St. Louis, Missouri (USA), were surveyed for exposure to canine distemper virus (CDV), canine adenovirus 1 (CAV-1); feline parvovirus (FPV; =feline panleukopenia), and several serovars of Leptospira interrogans. Exposure to each of the viruses and two Leptospira serovars (grippotyphosa and icterohemorrhagiae) was detected (prevalence of CDV = 54.1%; FPV = 49.7%; CAV-1 = 6.9%; L. interrogans icterohemorrhagiae = 8.9%; L. interrogans grippotyphosa = 6.3%). Eighty percent of raccoons showed evidence of exposure to at least one of the five primary pathogens, and 39% were positive for multiple species. Among the viruses, there was a significant co-occurrence of CDV and CAV-1. Longitudinal data on a subset of animals revealed that among individuals who were diagnosed as seropositive on first capture, 33-100% became seronegative for the pathogen of interest when reexamined at a later date. Thus, free-ranging urban raccoons have been exposed to multiple infectious agents, some of which may pose risks to humans and to nonvaccinated domestic and captive animal populations.


title: Laparoscopic Roux-en-Y gastric bypass in super obese Göttingen minipigs.

text: BACKGROUND The specific mechanisms behind weight loss and comorbidity improvements in obese patients after Roux-en-Y gastric bypass (RYGBP) are still poorly understood. The aim of this study was to establish and evaluate the feasibility of a long-term survival RYGBP model in super obese Göttingen minipigs in order to improve the translational potential relative to current animal models. METHODS Eleven Göttingen minipigs with diet-induced obesity underwent laparoscopic RYGBP and were followed up to 9 months after surgery. Intra- and post-operative complications, body weight (BW), food intake and necropsy data were recorded. RESULTS Five minipigs survived without complications to the end of the study. Four minipigs developed surgical related complications and were euthanized while two minipigs died due to central venous catheter related complications. BW and food intake is reported for the six minipigs surviving longer than 4.5 months post-surgery. Weight loss and reduced food intake was seen in all minipigs. After 2-3 months of weight loss, weight regain was evident in all but two minipigs which seemed to continue losing weight. Necropsy revealed some variation in the length of the alimentary, biliary and common limb between minipigs. CONCLUSION The use of obese Göttingen minipigs as a translational RYGBP model is feasible and has potential for the study of RYGBP-related changes in gut function, type-2 diabetes and appetite regulation. Still, the surgical procedure is technically highly demanding in obese Göttingen minipigs and the peri-operative animal care and follow up requires close monitoring.


title: Induction of apoptosis by the severe acute respiratory syndrome coronavirus 7a protein is dependent on its interaction with the Bcl-XL protein.

text: The severe acute respiratory syndrome coronavirus (SARS-CoV) 7a protein, which is not expressed by other known coronaviruses, can induce apoptosis in various cell lines. In this study, we show that the overexpression of Bcl-XL, a prosurvival member of the Bcl-2 family, blocks 7a-induced apoptosis, suggesting that the mechanism for apoptosis induction by 7a is at the level of or upstream from the Bcl-2 family. Coimmunoprecipitation experiments showed that 7a interacts with Bcl-XL and other prosurvival proteins (Bcl-2, Bcl-w, Mcl-1, and A1) but not with the proapoptotic proteins (Bax, Bak, Bad, and Bid). A good correlation between the abilities of 7a deletion mutants to induce apoptosis and to interact with Bcl-XL was observed, suggesting that 7a triggers apoptosis by interfering directly with the prosurvival function of Bcl-XL. Interestingly, amino acids 224 and 225 within the C-terminal transmembrane domain of Bcl-XL are essential for the interaction with the 7a protein, although the BH3 domain of Bcl-XL also contributes to this interaction. In addition, fractionation experiments showed that 7a colocalized with Bcl-XL at the endoplasmic reticulum as well as the mitochondria, suggesting that they may form complexes in different membranous compartments.


title: Immunological detection of severe acute respiratory syndrome coronavirus by monoclonal antibodies.

text: In order to establish immunological detection methods for severe acute respiratory syndrome coronavirus (SARS-CoV), we established monoclonal antibodies directed against structural components of the virus. B cell hybridomas were generated from mice that were hyper-immunized with inactivated SARS-CoV virion. By screening 2,880 generated hybridomas, we established three hybridoma clones that secreted antibodies specific for nucleocapsid protein (N) and 27 clones that secreted antibodies specific for spike protein (S). Among these, four S-protein specific antibodies had in vitro neutralization activity against SARS-CoV infection. These monoclonal antibodies enabled the immunological detection of SARS-CoV by immunofluorescence staining, Western blot or immunohistology. Furthermore, a combination of monoclonal antibodies with different specificities allowed the establishment of a highly sensitive antigen-capture sandwich ELISA system. These monoclonal antibodies would be a useful tool for rapid and specific diagnosis of SARS and also for possible antibody-based treatment of the disease.


title: Travel health nursing: expanding horizons for occupational health nurses.

text: Pre-travel care, with its focus on disease prevention and health promotion, is the cornerstone of travel health nursing practice. Pre-travel nursing services include risk assessment, immunizations, health counseling, and referrals. Individual travel health and safety risks vary and depend upon a number of different factors. These include: traveler health status, destination, trip duration, season of travel, type of accommodation, trip activities, and prevention efforts. More than 35% of U.S. international travel is business travel. Occupational health nurses have an important role to play to keep traveling employees, including frequent flyers, long stay expatriates, and special needs travelers, healthy, safe, and productive in the "global marketplace."


title: Surveillance of avian influenza A(H7N9) virus infection in humans and detection of the first imported human case in Taiwan, 3 April to 10 May 2013.

text: On 3 April 2013, suspected and confirmed cases of influenza A(H7N9) virus infection became notifiable in the primary care sector in Taiwan, and detection of the virus became part of the surveillance of severe community-acquired pneumonia. On 24 April, the first imported case, reported through both surveillance systems, was confirmed in a man returning from China by sequencing from endotracheal aspirates after two negative throat swabs. Three of 139 contacts were ill and tested influenza A(H7N9)-negative.


title: Maternal pertussis is hazardous for a newborn: a case report.

text: Pertussis, or whooping cough, a highly contagious disease caused by Bordetella pertussis, is making a comeback globally and nationally in spite of reasonable vaccination coverage. Worldwide, there have been increasing reports of Bordetella pertussis infection among adolescents and adults, but the peak incidence and highest mortality occur among infants. We report a 19-day-old female infant presenting with progressive respiratory failure. The mother was the only familial contact who complained of mild cough. However, occasional apneic episodes with cyanosis and peripheral lymphocytosis prompted us to examine the presence of Bordetella pertussis, which remains a significant cause of morbidity and mortality in unimmunized infants. Understanding the source of pertussis transmission to infants may provide new approaches to prevent pertussis in the most vulnerable infants. Various potential strategies have been reviewed or recommended in countries with the aim of better protecting infants against pertussis. Public health measures to prevent the disease could be strengthened and booster vaccinations against pertussis considered.


title: [Experience with empirical treatment of severe acute respiratory syndrome due to coronavirus, genotype IV].

text: Foreign experience with empirical management of severe acute respiratory syndrome (SARS) due to coronavirus, genotype IV is described. It is indicated that the data on the efficacy of ribavirin with respect to SARS in the patients are contradictory. The efficacy of two chemotherapeutics, i. e. lopinavir and ritonavir used in the SARS foci is confirmed. The drugs are at present applicable all over the world in retrovirus therapy of HIV infected subjects. The search for efficient Russian unspecific medicines for control of SARS is actual.


title: Cohort Study of the Impact of High-Dose Opioid Analgesics on Overdose Mortality.

text: OBJECTIVE Previous studies examining opioid dose and overdose risk provide limited granularity by milligram strength and instead rely on thresholds. We quantify dose-dependent overdose mortality over a large spectrum of clinically common doses. We also examine the contributions of benzodiazepines and extended release opioid formulations to mortality. DESIGN Prospective observational cohort with one year follow-up. SETTING One year in one state (NC) using a controlled substances prescription monitoring program, with name-linked mortality data. SUBJECTS Residential population of North Carolina (n = 9,560,234), with 2,182,374 opioid analgesic patients. METHODS Exposure was dispensed prescriptions of solid oral and transdermal opioid analgesics; person-years calculated using intent-to-treat principles. Outcome was overdose deaths involving opioid analgesics in a primary or additive role. Poisson models were created, implemented using generalized estimating equations. RESULTS Opioid analgesics were dispensed to 22.8% of residents. Among licensed clinicians, 89.6% prescribed opioid analgesics, and 40.0% prescribed ER formulations. There were 629 overdose deaths, half of which had an opioid analgesic prescription active on the day of death. Of 2,182,374 patients prescribed opioids, 478 overdose deaths were reported (0.022% per year). Mortality rates increased gradually across the range of average daily milligrams of morphine equivalents. 80.0% of opioid analgesic patients also received benzodiazepines. Rates of overdose death among those co-dispensed benzodiazepines and opioid analgesics were ten times higher (7.0 per 10,000 person-years, 95 percent CI: 6.3, 7.8) than opioid analgesics alone (0.7 per 10,000 person years, 95 percent CI: 0.6, 0.9). CONCLUSIONS Dose-dependent opioid overdose risk among patients increased gradually and did not show evidence of a distinct risk threshold. There is urgent need for guidance about combined classes of medicines to facilitate a better balance between pain relief and overdose risk.


title: Antibodies to selected pathogens in free-ranging terrestrial carnivores and marine mammals in Canada.

text: Antibody titres to selected pathogens (canine adenovirus [CAV-2], feline herpesvirus [FHV], phocine herpesvirus [PHV-1], canine distemper virus, dolphin morbillivirus [DMV], phocine distemper virus [PDV], parainfluenza virus type 3 [PI3], rabies virus, dolphin rhabdovirus [DRV], canine coronavirus, feline coronavirus, feline leukaemia virus, Borrelia burgdorferi and Toxoplasma gondii) were determined in whole blood or serum samples from selected free-ranging terrestrial carnivores and marine mammals, including cougars (Fellis concolor), lynxes (Fellis lynx), American badgers (Taxidea taxus), fishers (Martes pennanti), wolverines (Gulo gulo), wolves (Canis lupus), black bears (Ursus americanus), grizzly bears (Ursus arctos), polar bears (Ursus maritimus), walruses (Odobenus rosmarus) and belugas (Delphinapterus leucas), which had been collected at several locations in Canada between 1984 and 2001. Antibodies to a number of viruses were detected in species in which these infections have not been reported before, for example, antibodies to CAV-2 in walruses, to PDV in black bears, grizzly bears, polar bears, lynxes and wolves, to DMV in grizzly bears, polar bears, walruses and wolves, to PI3 in black bears and fishers, and to DRV in belugas and walruses.


title: [Severe acute respiratory syndrome].

text: OBJECTIVE Severe acute respiratory syndrome(SARS), caused by SARS- associated coronavirus(SCV), is the first severe infectious disease in this century. SARS is pathologically characterized by interstitial exudative inflammation of lung with the formation of hyaline membrane in acute phase. Haemorrhagic inflammation exists in extrapulmonary organs. Clinical diagnosis is a dynamic process and includes the suspected case, probable case and definite case. Diagnostic standard of SARS will be revised with further understanding of the disease. Chinese term of SARS has been recommended in the paper.


title: The predicted metal-binding region of the arterivirus helicase protein is involved in subgenomic mRNA synthesis, genome replication, and virion biogenesis.

text: Equine arteritis virus (EAV), the prototype Arterivirus, is a positive-stranded RNA virus that expresses its replicase in the form of two large polyproteins of 1,727 and 3,175 amino acids. The functional replicase subunits (nonstructural proteins), which drive EAV genome replication and subgenomic mRNA transcription, are generated by extensive proteolytic processing. Subgenomic mRNA transcription involves an unusual discontinuous step and generates the mRNAs for structural protein expression. Previously, the phenotype of mutant EAV030F, which carries a single replicase point mutation (Ser-2429-->Pro), had implicated the nsp10 replicase subunit (51 kDa) in viral RNA synthesis, and in particular in subgenomic mRNA transcription. nsp10 contains an N-terminal (putative) metal-binding domain (MBD), located just upstream of the Ser-2429-->Pro mutation, and a helicase activity in its C-terminal part. We have now analyzed the N-terminal domain of nsp10 in considerable detail. A total of 38 mutants, most of them carrying specific single point mutations, were tested in the context of an EAV infectious cDNA clone. Variable effects on viral genome replication and subgenomic mRNA transcription were observed. In general, our results indicated that the MBD region, and in particular a set of 13 conserved Cys and His residues that are assumed to be involved in zinc binding, is essential for viral RNA synthesis. On the basis of these data and comparative sequence analyses, we postulate that the MBD may employ a rather unusual mode of zinc binding that could result in the association of up to four zinc cations with this domain. The region containing residue Ser-2429 may play the role of "hinge spacer," which connects the MBD to the rest of nsp10. Several mutations in this region specifically affected subgenomic mRNA synthesis. Furthermore, one of the MBD mutants was replication and transcription competent but did not produce infectious progeny virus. This suggests that nsp10 is involved in an as yet unidentified step of virion biogenesis.


title: S1 and N gene analysis of avian infectious bronchitis viruses in Taiwan.

text: The disease caused by infectious bronchitis virus (IBV) produces great economic for the poultry industry. The purpose of this study is to investigate the molecular epidemiology of IBV in Taiwan. An old IBV strain isolated in 1964 and another 31 strains isolated from 1991 to 2003 were selected for N-terminal S1 gene analysis. Based on their phylogenetic tree, 13 strains were selected for sequencing the entire S1 and partial nucleocapsid (N) genes. The results indicated that Taiwanese IBV strains could be divided into two distinct lineages, Taiwan Group I and Taiwan Group II, with one Massachusetts strain and one Chinese strain. No recombination was found between H120 and the Taiwanese strains in the S1 gene. However, the S1 gene showed a noticeably higher divergence than the N gene. The phylogenetic trees constructed from the S1 and N genes indicate that intergenic recombination has occurred. Since most local strains are in Taiwanese clusters, developing vaccines from local strains is necessary for IBV control in Taiwan.


New drugs for Clostridium difficile infection.


title: Infection of retinal neurons during murine cytomegalovirus retinitis.

text: PURPOSE Previous results suggest that retinal neurons are infected early during murine cytomegalovirus (MCMV) infection of the inner retina. The purposes of this study were to identify which retinal neurons are infected and to determine the routes by which MCMV spreads in the retina. METHODS Immunosuppressed (IS) BALB/c mice were inoculated with 5 x 10(3) PFU of MCMV (k181) through the supraciliary route. Injected eyes were collected at several times after inoculation, sectioned, and examined by electron microscopy and by staining for retinal cell antigens and for MCMV early (EA) or late (LA) antigen. RESULTS MCMV-infected cells were observed in the choroid and RPE by day 3 after infection (PI) and in the inner retina beginning at day 5 PI. At this time, many horizontal and bipolar cells were MCMV-antigen-positive but only rare MCMV-infected amacrine cells (glycine positive or gamma-aminobutyric acid [GABA] positive) or MCMV-infected ganglion cells (NF positive) were observed in the inner retina. At day 10 PI, most virus-infected cells were glial fibrillary acidic protein (GFAP)- and GABA-positive glia. Virions were observed by electron microscopy in the choroid, RPE, and inner nuclear layer of the retina. Although virions were observed in the endothelium of the retinal vessels and the nearby retinal cells, the endothelial cell lining of the retinal vessels remained intact. Both apoptotic cells and necrotic cells were seen in the inner retina. CONCLUSIONS In the inner retina, horizontal and bipolar cells were the early (< or = day 7 PI) targets of MCMV infection. Virus spread from the RPE and the photoreceptor layer to the inner retina through infected Muller cells and within the inner retina horizontally through infected horizontal cells.


Neonatal intensive care and parental participation in decision making.


title: Psychological distress and negative appraisals in survivors of severe acute respiratory syndrome (SARS).

text: BACKGROUND Severe acute respiratory syndrome (SARS) is a novel disease. The authors have limited knowledge of its impact on mental health. The present study aimed to examine the level and extent of psychological distress of SARS survivors following 1-month recovery, to explore patients' negative appraisals of the impact of SARS, and to evaluate the associations between psychological distress and negative appraisals. METHOD The Beck Anxiety Inventory, the Beck Depression Inventory, and a newly developed measure, the SARS Impact Scale (SIS), were mailed to 453 Hong Kong Chinese SARS survivors discharged from hospital for 4 weeks or more. RESULTS A total of 425 patients received the questionnaires and 180 (mean age 36.9 years; 120 women) gave valid replies. The response rate was 42.4 %. The participants also represented 13.6 % of all adult survivors in Hong Kong. About 35 % of respondents reported 'moderate to severe' or 'severe' ranges of anxiety and/or depressive symptoms. It was found that those working as healthcare workers or having family members killed by SARS were more prone to develop subsequent high levels of distress. Factor analyses extracted three meaningful factors of the SIS, namely 'survival threat', 'physical impact', and 'social impact'. Negative appraisals at the acute phase and 1-month recovery significantly accounted for substantial portions of variances for anxiety and depressive symptoms, after the effects of other psychosocial variables were controlled. CONCLUSIONS Psychological distress of SARS survivors at 1-month recovery is real and significant. Negative appraisals may play a pivotal role in the development of psychological distress for SARS survivors, at least in the short term.


title: [The immunological and molecular diagnostics of respiratory tract infections].

text: Diagnostic strategies in upper and lower respiratory tract infections include conventional procedures and the new molecular tools. Serology plays a role in certain viral and atypical infections of lower respiratory track but is not reliable in small children and in immunocompromised subjects. Antigen detection can be also carried out. If a potential pathogen is non detectable by simply assay or in severe or complicated lower respiratory tract infections, the use of molecular tools is highly indicated.


title: Serologic diagnosis of canine leishmaniasis by dot-ELISA.

text: A dot-enzyme-linked immunosorbent assay (ELISA) using protein A-peroxidase was evaluated as a diagnostic test for canine leishmaniasis. The test results were in agreement with parasitologic diagnosis and indirect immunofluorescence assay results. The sensitivity of the test calculated on 31 dogs with positive parasitologic examination was 90% when a titer of 1/800 was established as a cutoff and 100% when a titer of 1/400 was established. The specificity calculated on the canine population from nonendemic areas was 100% when both titers were established. Nevertheless, in endemic areas titers near the cutoff need careful interpretation. The results of this study demonstrate that dot-ELISA protein A using a bio-dot apparatus is highly suitable for seroepidemiologic field work.