title: Effect of single treatment with the antihypertensive drug eplerenone on hormone levels and anxiety-like behaviour in rats.

text: OBJECTIVE The aim of the study was to investigate the effects of single peripheral administration of mineralocorticoid receptor (MR) antagonist eplerenone on: 1. the hypothalamic-pituitary-adrenocortical (HPA) axis; 2. the renin-angiotensin-aldosterone (RAA) system, 3. anxiety-like behaviour. METHODS Male Wistar rats were injected subcutaneously with eplerenone (100 mg/kg body weight) or vehicle. Two hours following the injections, a half of the animals from each treatment group were decapitated to obtain blood for measurement of hormone levels. The other half of the animals was subjected to behavioural testing in the elevated plus-maze test. To provide comprehensive behavioural profile of eplerenone, standard spatiotemporal and ethologically derived measures of anxiety were assessed. RESULTS Single treatment with eplerenone resulted in a significant increase in plasma aldosterone levels. Plasma concentrations of ACTH and corticosterone were not modified by eplerenone injection. Administration of eplerenone failed to alter classical spatiotemporal measures of anxiety (number of entries and time spent in the open arms). However, ethological parameters related to exploration (head dipping) and risk assessment behaviour (stretched attend postures) were significantly affected by eplerenone injection. CONCLUSIONS Single injection of eplerenone is followed by a reduction of ethological indices of anxiety-like behaviour and by an elevation of plasma aldosterone levels. Acute administration of eplerenone appears to affect the RAA system but not hormones of the HPA axis.


title: [Clinical features of patients with influenza-like illness who went to a third level center in the winter of 2013-2014].

text: BACKGROUND Between October 2013 and April 2014, the Centers for Disease Control and Prevention reported a high incidence of AH1N1 influenza cases. Not all suspicious cases of influenza are confirmed; however, all patients need treatment and generate additional costs to the institutions. Our objective was to describe the characteristics of the patients treated for influenza suspicion at Hospital de Especialidades in Centro Médico Nacional Siglo XXI during a new epidemic. METHODS Cross-sectional study of all cases admitted at the Emergency Service (of the aforementioned hospital) for influenza-like illness, defined according to current guidelines. Based on these guidelines, we analyzed the risk factors that may have increased the severity of the infection. RESULTS We registered a total of 109 patients with a mean age of 44 years, 78 % were under 60 years of age, 62 % were women, 75 % had at least one comorbidity, such as obesity (26 %) or hypertension (27 %). Only 65 patients had results from a confirmatory test, 33.8 % had positive diagnosis, and 21 % of them eventually died. CONCLUSIONS The frequency of confirmed cases for influenza infection is low. The risk factors associated with complications and increased mortality are hypertension, leukocytosis and clinical presentation of severe acute respiratory syndrome.


docid: mvmczbnk

title: Structural characterization of the SARS-coronavirus spike S fusion protein core.

text: The spike (S) glycoprotein of coronaviruses mediates viral entry into host cells. It is a type 1 viral fusion protein that characteristically contains two heptad repeat regions, denoted HR-N and HR-C, that form coiled-coil structures within the ectodomain of the protein. Previous studies have shown that the two heptad repeat regions can undergo a conformational change from their native state to a 6-helix bundle (trimer of dimers), which mediates fusion of viral and host cell membranes. Here we describe the biophysical analysis of the two predicted heptad repeat regions within the severe acute respiratory syndrome coronavirus S protein. Our results show that in isolation the HR-N region forms a stable alpha-helical coiled coil that associates in a tetrameric state. The HR-C region in isolation formed a weakly stable trimeric coiled coil. When mixed together, the two peptide regions (HR-N and HR-C) associated to form a very stable alpha-helical 6-stranded structure (trimer of heterodimers). Systematic peptide mapping showed that the site of interaction between the HR-N and HR-C regions is between residues 916-950 of HR-N and residues 1151-1185 of HR-C. Additionally, interchain disulfide bridge experiments showed that the relative orientation of the HR-N and HR-C helices in the complex was antiparallel. Overall, the structure of the hetero-stranded complex is consistent with the structures observed for other type 1 viral fusion proteins in their fusion-competent state.


title: Molecular approach to find target(s) for oligoclonal bands in multiple sclerosis.

text: OBJECTIVES Oligoclonal bands are a characteristic finding in the CSF of patients with multiple sclerosis, yet their target antigen(s) remain unknown. The objective was to determine whether a filamentous phage peptide library could be employed to allow the oligoclonal bands to select their own target epitopes. METHODS CSF IgG antibody from 14 patients with multiple sclerosis and 14 controls was used to select individual phage clones from a bacteriophage library containing approximately 4 x 10(7) different hexamers expressed on its surface pIII protein. The amino acid sequence selected was deduced by sequencing the DNA of the genetically engineered insert. RESULTS In general, after three rounds of selection, CSF from both patients with multiple sclerosis and controls selected one to two consistent peptide motifs. Five out of 14 patients with multiple sclerosis, and one control, selected the amino acid sequence motif, RRPFF. Given 20 possible amino acids per position, the likelihood of five patients selecting the same linear five amino acid sequence is at most 1.6 x 10(-3), corrected for the number of clones sequenced. A GenBank computer search showed that this sequence is found in the Epstein-Barr Virus nuclear antigen (EBNA-1), and a heat shock protein alphaB crystallin. Human serum antibodies to a synthetic peptide containing RRPFF were virtually exclusively found in patients with prior infection by Epstein-Barr virus. Other studies have suggested a relation between Epstein-Barr virus infection and multiple sclerosis, including nearly 100% Epstein-Barr virus seropositivity among patients with multiple sclerosis and increased concentrations of antibody to EBNA in CSF of patients with multiple sclerosis. By antigen specific immunoblotting, antibodies to the RRPFF motif in the CSF were shown to correspond to a subset of oligoclonal bands in the CSF from the same patient. CONCLUSION This study shows that phage epitope display libraries may be used to select amino acid motifs which are potentially relevant to the pathogenesis of multiple sclerosis.


title: [The virtual ward - the students' clinical education during the current pandemic].

text: On March 17, 2020, the Swedish Government recommended all higher education institutions to move to online and distance learning during the COVID-19 pandemic. The integrated course in Infection, Microbiology, and Immunity at the Programme in Medicine at University of Gothenburg had to be completely transformed. Creative solutions have now replaced the clinical training that normally takes place during the students' clinical education at the hospital. We developed a digital concept entitled "the virtual ward", in which we interact with the students in real time. Here, the students are able to follow their patients on a daily basis during teacher-guided sessions.


title: Coronavirus JHM: coding assignments of subgenomic mRNAs.

text: Protein synthesis in the murine hepatitis virus JHM-infected cells was temporarily inhibited by hypertonic shock. When the cells were returned to isotonic medium the synthesis of six virus-specific polypeptides, 150K, 65K, 60K, 30K, 23K and 14K was reinitiated simultaneously. Polyadenylated RNA isolated from the cytoplasm or polysomes of infected cells was translated in vitro and the products included polypeptides with molecular weights (mol. wt.) of 120,000, 60,000, 30,000, 23,000 and 14,000. Immunoprecipitation and fingerprinting of [35S]methionine-containing tryptic peptides showed that the 60,000 and 23,000 mol. wt. products were identical to the 60K and 23K polypeptides found in infected cells; the 120,000 mol. wt. product showed identity with the 150K intracellular polypeptide and a virus-specific 120K polypeptide synthesized in tunicamycin-treated cells. Two-dimensional polyacrylamide gel electrophoresis strongly suggested that the 30,000 and 14,000 mol. wt. products are equivalent to virus-specific 30K and 14K intracellular polypeptides. [3H]Uridine-labelled polyadenylated virus RNA was isolated from infected cells and sedimented in sucrose gradients containing formamide. The distribution in the gradient of each of the previously identified virus RNAs was determined by gel electrophoresis and gradient fractions enriched for each RNA were translated in vitro. The 120,000, 60,000, 30,000, 23,000 and 14,000 mol. wt. polypeptides were found to be encoded by mRNAs 3, 7, 2, 6, and 4 or 5 respectively. These results demonstrate that the virus-specific polypeptides in JHM-infected cells are encoded in separate subgenomic mRNAs and are translated independently. The assignment of coding functions and the known sequence relationships of JHM RNAs permitted a gene order to be deduced.


title: Covid 19 pandemic and gynaecological laparoscopic surgery: knowns and unknowns.

text: The worldwide impact of COVID 19 continues to be felt as hospitals in all countries reduce elective and non-urgent cases to allow staffing and resources to be deployed elsewhere. Urgent gynaecological and cancer procedures are continuing, and it is imperative all theatre staff are protected and risks of SARS-CoV-2 viral transmission reduced when operating on asymptomatic, suspected or confirmed COVID 19 patients. In particular, there are concerns relating to the transmission of COVID 19 during gynaecological laparoscopic surgery, arising from the potential generation of SARS-CoV-2 contaminated aerosols from CO2 leakage and the creation of smoke from the use of energy devices. The aim of this paper is to review all the up to date evidence, including experiences from China and Italy, to guide the safe management of such patients when undergoing gynaecological procedures.


title: Impact of a spreading epidemic on medical students.

text: The emergence of severe acute respiratory syndrome (SARS) had caused fear and anxiety of unprecedented proportion. To examine the impact of SARS on the medical students in a private medical university, a self-reporting questionnaire study was carried out to assess the factual knowledge, anxiety level and perception of the crisis, among the students. The two-week study (between 12 and 23 May, 2003) was carried out three weeks after the first reported SARS-related death in Malaysia. Ninety-one Phase I (junior) and 113 Phase II (senior) students completed the questionnaires. A large majority of students of Phase I and II were correct in their factual knowledge and were sensible in their perception of the future and the handling of the crisis by government(s). However, phase 1 students expressed significantly greater degree of anxiety compared to Phase II in relation to attendance and personal protection in hospital, and in meeting people coughing in public places. The lesser degree of anxiety expressed by phase II senior students may be due in part, to a more realistic assessment of SARS risk brought about by maturity, time spent in hospital and interaction with clinical lecturers and medical staff.


title: A tale of two cities: community psychobehavioral surveillance and related impact on outbreak control in Hong Kong and Singapore during the severe acute respiratory syndrome epidemic.

text: OBJECTIVES To compare the public's knowledge and perception of SARS and the extent to which various precautionary measures were adopted in Hong Kong and Singapore. DESIGN Cross-sectional telephone survey of 705 Hong Kong and 1,201 Singapore adults selected by random-digit dialing. RESULTS Hong Kong respondents had significantly higher anxiety than Singapore respondents (State Trait Anxiety Inventory [STAI] score, 2.06 vs 1.77; P < .001). The former group also reported more frequent headaches, difficulty breathing, dizziness, rhinorrhea, and sore throat. More than 90% in both cities were willing to be quarantined if they had close contact with a SARS case, and 70% or more would be compliant for social contacts. Most respondents (86.7% in Hong Kong vs 71.4% in Singapore; P < .001) knew that SARS could be transmitted via respiratory droplets, although fewer (75.8% in Hong Kong vs 62.1% in Singapore; P < .001) knew that fomites were also a possible transmission source. Twenty-three percent of Hong Kong and 11.9% of Singapore respondents believed that they were "very likely" or "somewhat likely" to contract SARS during the current outbreak (P < .001). There were large differences between Hong Kong and Singapore in the adoption of personal precautionary measures. Respondents with higher levels of anxiety, better knowledge about SARS, and greater risk perceptions were more likely to take comprehensive precautionary measures against the infection, as were older, female, and more educated individuals. CONCLUSION Comparative psychobehavioral surveillance and analysis could yield important insights into generic versus population-specific issues that could be used to inform, design, and evaluate public health infection control policy measures.


title: Development of one-step, real-time, quantitative reverse transcriptase PCR assays for absolute quantitation of human coronaviruses OC43 and 229E.

text: The clinical significance of human coronaviruses in more severe respiratory illnesses has recently been shown to be higher than was previously assumed. Rapid and reliable diagnosis of human coronavirus infections therefore becomes indispensable in a routine clinical setting. In this study, we present a very sensitive and specific TaqMan-based, real-time quantitative reverse transcriptase PCR (qRT-PCR) for the rapid detection and quantitation of human coronaviruses (HCoVs) OC43 and 229E. Absolute viral load measurement in clinical samples was achieved through the construction of in-house HCoV OC43 and 229E cRNA standards for the generation of a standard curve. The HCoV OC43 assay allows quantitation over a range from 20 to 2 x 10(8) RNA copies per reaction mixture (5 microl RNA extract). When this is extrapolated to clinical samples, this corresponds to a detection range of 10(3) to 10(10) viral genome equivalents per ml. By using the HCoV 229E qRT-PCR assay, viral RNA copies ranging from 200 to 2 x 10(9) per reaction mixture can be detected, which corresponds to 10(4) to 10(11) viral genome equivalents per ml sample. A total of 100 respiratory samples screened for the presence of HCoVs OC43 and 229E by using conventional RT-PCR were assessed in parallel by the qRT-PCR assays. By use of the real-time qRT-PCR techniques, the detection rate of HCoVs OC43 and 229E increased from 2.0% to 3.1% and from 0.3% to 2.5%, respectively. The real-time qRT-PCR assays described here allow the rapid, specific, and sensitive laboratory detection and quantitation of human coronaviruses OC43 and 229E.


title: Screening of respiratory pathogens by Respiratory Multi Well System (MWS) r-gene™ assay in hospitalized patients.

text: Novel respiratory viruses have been identified as possible agents of upper and lower respiratory tract infections. Multiplex real-time PCRs have been developed to identify clinically relevant respiratory pathogens. In this study, 178 respiratory samples already screened for influenza virus types A and B by Flu A/B ASR real-time PCR kit were retrospectively analyzed with the Respiratory Multi Well System (MWS) r-gene™ real-time PCR kit which detects a wide spectrum of respiratory pathogens. The goal was to demonstrate the importance of a wide spectrum screening compared to a single diagnostic request. The Flu A/B ASR kit detected influenza B virus in 1.7% of the samples (3/178) and no influenza A virus. The MWS r-gene™ kit detected influenza virus in 6.7% (12/178) of samples (0.6% influenza A, and 6.2% influenza B), while the overall detection rate for respiratory pathogens was 54% (96/178). Co-infections were detected in 8/178 (4.5%) samples. Adenovirus was the infectious agent detected most frequently, followed by respiratory syncytial virus. The risk of being infected by respiratory syncytial virus is almost threefold higher in patients older than 65 years compared to the younger age group (OR:2.7, 95% CI: 1.2-6.2). Wide spectrum screening of respiratory pathogens by real-time PCR is an effective means of detecting clinically relevant viral pathogens.


title: Novel aspects of the renin-angiotensin-aldosterone-system.

text: The renin-angiotensin-aldosterone system (RAAS) play a pivotal role in the progression of renal disease. The RAAS has become much more complex in recent years with the identification of novel peptides that exhibit biological activity. There are novel pathways of angiotensin II (ANG II) generation independent of angiotensin converting enzyme (ACE). ANG II bind to at least two different receptors and prorenin/renin also exerts pathophysiological effects through binding to specific receptor. ANG II itself has emerged as a multifunctional cytokine exhibiting many non-hemodynamic properties such as acting as a growth factor and profibrogenic and proinflammatory cytokine. These profibrogenic and proinflammatory effects are mediated by other factors such as transforming growth-factor beta (TGF-beta) and chemoattractants that are induced in the kidney by ANG II. Increased aldosterone levels contribute to renal injury, independent of blood pressure or ANG II. Numerous experimental and clinical studies have shown that ACE-inhibitors as well as AT1-receptor antagonists can prevent glomerulosclerosis and tubulointerstitial fibrosis. This review will highlight some of these novel insights into the RAAS in regards to renal injury.


title: Echocardiographic assessment of coronary artery flow in normal canines and model dogs with myocardial infarction.

text: This study was conducted to evaluate the usefulness of coronary arterial profiles from normal dogs (11 animals) and canines (six dogs) with experimental myocardial infarction (MI) induced by ligation of the left coronary artery (LCA). Blood velocity of the LCA and right coronary artery (RCA) were evaluated following transthoracic pulsed-wave Doppler echocardiography. The LCA was observed as an infundibular shape, located adjacent to the sinus of Valsalva. The RCA appeared as a tubular structure located 12 o'clock relative to the aorta. In normal dogs, the LCA and RCA mean peak diastolic velocities were 20.84 ± 3.24 and 19.47 ± 2.67 cm/sec, respectively. The LCA and RCA mean diastolic deceleration times were 0.91 ± 0.14 sec and 1.13 ± 0.20 sec, respectively. In dogs with MI, the LCA had significantly (p < 0.01) lower peak velocities (14.82 ± 1.61 cm/sec) than the RCA (31.61 ± 2.34 cm/sec). The RCA had a significantly (p < 0.01) rapid diastolic deceleration time (0.71 ± 0.06 sec) than that found in the LCA (1.02 ± 0.22 sec) of MI dogs. In conclusion, these profiles may serve as a differential factor for evaluating cardiomyopathy in dogs.


title: Factors that influence Medical Reserve Corps recruitment.

text: The Medical Reserve Corps (MRC) is a key strategy used in the United States to assure an adequate surge capacity healthcare workforce for response to disasters. A survey of Hawaiian healthcare providers (n = 1,057) was conducted to identify factors that influence interest, ability, and willingness to join the MRC; 468 (44.3%) healthcare providers responded. Overall, females were more likely to demonstrate an interest in joining the MRC, while physicians and dentists reported lower levels of ability and willingness, in addition to a lower level of interest in joining the MRC than the other professional groups. The most important motivating factor in joining the MRC was altruism and the ability to help one's own community. Respondents reported a number of factors that would influence their decision to join or remain a MRC member. These included: (1) time commitment required; (2) MRC organization and management; (3) provision of MRC-sponsored training or education sessions and continuing education credits; (4) concerns regarding the safety of family members during a disaster; (5) professional liability protection for work performed during MRC operations; and (6) competing personal obligations. Strategies targeting these factors probably will be most effective in recruitment and retention of MRC volunteers as well as members of other public health surge capacity volunteer groups.


title: [Detection of coronavirus in experimental pneumocystic pneumonia].

text: Transmission electron microscopy (of ultrathin sections) was used to examine the biomass of lung tissue in the immunodeficiency minipigs experimentally infected with Pneumocystis carinii. The material was found to contain pneumocysts, bacteria, and coronaviruses. There was a clear coronavirus-pneumocyst structural relationship. The findings suggest the combined effect of microorganisms of different systematic groups on the development of a pathological process in the experimental infection etiologically determined by Pneumocystis carinii.


title: The severe acute respiratory syndrome coronavirus Nsp15 protein is an endoribonuclease that prefers manganese as a cofactor.

text: Nonstructural protein 15 (Nsp15) of the severe acute respiratory syndrome coronavirus (SARS-CoV) produced in Escherichia coli has endoribonuclease activity that preferentially cleaved 5' of uridylates of RNAs. Blocking either the 5' or 3' terminus did not affect cleavage. Double- and single-stranded RNAs were both substrates for Nsp15 but with different kinetics for cleavage. Mn(2+) at 2 to 10 mM was needed for optimal endoribonuclease activity, but Mg(2+) and several other divalent metals were capable of supporting only a low level of activity. Concentrations of Mn(2+) needed for endoribonuclease activity induced significant conformation change(s) in the protein, as measured by changes in tryptophan fluorescence. A similar endoribonucleolytic activity was detected for the orthologous protein from another coronavirus, demonstrating that the endoribonuclease activity of Nsp15 may be common to coronaviruses. This work presents an initial biochemical characterization of a novel coronavirus endoribonuclease.


title: Genetic diversity of avian infectious bronchitis virus isolated from domestic chicken flocks and coronaviruses from feral pigeons in Brazil between 2003 and 2009.

text: To detect the presence of infectious bronchitis virus or avian coronavirus, a nested reverse transcriptase PCR (RT-PCR) method was developed with the aim of amplifying a fragment of 530 bases, comprising the gene coding S1 protein. In the first step, all samples were submitted to RNA extraction, RT-PCR, and nested PCR. Next, only the positive nested-PCR samples were propagated in specific-pathogen-free (SPF) embryonated chicken eggs for virus isolation. Positive samples were then sequenced and analyzed using a molecular phylogeny approach. Tracheal swab samples were collected from 23 different domestic chickens distributed in three regions of Brazil, in the period between 2003 and 2009. Also analyzed were six swab samples (tracheal and cloacal) from asymptomatic pigeons (Columba livia), caught in an urbanized region in southeastern Brazil. The study revealed two major phylogenetic groups: one clustered with the Massachusetts vaccine serotype and another joined with the D207 strain. Interestingly, samples grouped with the Connecticut and Arkansas serotypes were also found. Pigeon isolates clustered with the Massachusetts serotype showed significant similarity (close to 100%) to those obtained from chickens. Only one pigeon isolate was seen to be grouped with the Connecticut serotype, and no correlation was observed between sample grouping and region origin. Understanding the diversity of genotypes and eco-epizootiology of the disease in different environments is expected to be helpful for vaccine production aimed at the main circulating variants. In this respect, one could also expect benefits in the management of other bird species that may act as avian coronavirus reservoirs.


title: Mutational analysis of heptad repeats in the membrane-proximal region of Newcastle disease virus HN protein.

text: For most paramyxoviruses, syncytium formation requires the expression of both surface glycoproteins (HN and F) in the same cell, and evidence suggests that fusion involves a specific interaction between the HN and F proteins (X. Hu et al., J. Virol. 66:1528-1534, 1992). The stalk region of the Newcastle disease virus (NDV) HN protein has been implicated in both fusion promotion and virus specificity of that activity. The NDV F protein contains two heptad repeat motifs which have been shown by site-directed mutagenesis to be critical for fusion (R. Buckland et al., J. Gen. Virol. 73:1703-1707, 1992; T. Sergel-Germano et al., J. Virol. 68:7654-7658, 1994; J. Reitter et al., J. Virol. 69:5995-6004, 1995). Heptad repeat motifs mediate protein-protein interactions by enabling the formation of coiled coils. Upon analysis of the stalk region of the NDV HN protein, we identified two heptad repeats. Secondary structure analysis of these repeats suggested the potential for these regions to form alpha helices. To investigate the importance of this sequence motif for fusion promotion, we mutated the hydrophobic a-position amino acids of each heptad repeat to alanine or methionine. In addition, hydrophobic amino acids in other positions were also changed to alanine. Every mutant protein retained levels of attachment activity that was greater than or equal to the wild-type protein activity and bound to conformation-specific monoclonal as well as polyclonal antisera. Neuraminidase activity was variably affected. Every mutation, however, showed a dramatic decrease in fusion promotion activity. The phenotypes of these mutant proteins indicate that individual amino acids within the heptad repeat region of the stalk domain of the HN protein are important for the fusion promotion activity of the protein. These data are consistent with the idea that the HN protein associates with the F protein via specific interactions between the heptad repeat regions of both proteins.


title: Influenza vaccination and hospitalisation in Elderly Health Centres.

text: 1. A cohort of Elderly Health Centres was examined to determine whether influenza vaccination decreased hospitalisation and mortality. 2. In the influenza season, influenza vaccination reduced all-cause mortality by half and cardiorespiratory hospitalisation by a quarter. The extent to which influenza vaccination protects older people from serious morbidity and mortality needs to be confirmed in appropriately designed studies, so that scarce health care resources can be used effectively


title: Feline infectious peritonitis: a review of clinicopathological changes in 65 cases, and a critical assessment of their diagnostic value.

text: In 65 natural cases of feline infectious peritonitis (FIP) the common clinicopathological changes included lymphopenia (77 per cent), neutrophilia (45 per cent), anaemia (37 per cent), hyperproteinaemia (39 per cent) and hyperglobulinaemia (39 per cent). There was no difference in the frequency of these abnormalities between the 38 cases of effusive disease and the 27 cases of non-effusive disease. The most consistent changes shown by serum protein electrophoresis were increases in alpha 2- and gamma-globulins. The protein content of the effusions ranged from 39 to 98 g/litre with the globulins comprising 50 to 82 per cent. Coronavirus serology showed a wide variation in antibody titres (0 to 2560) with 320 the modal titre. The diagnostic value of this information was evaluated by comparing it with data from 65 cats in which FIP was considered as a differential diagnosis, but another disease was diagnosed. None of the laboratory tests, including coronavirus serology, had good sensitivity and specificity for the diagnosis of the disease. The presence of multiple abnormalities compatible with the disease increased the specificity but decreased the sensitivity of the diagnosis.


Avian infectious bronchitis virus: isolation of an apparently new variant in Italy.


title: Infection control measures for operative procedures in severe acute respiratory syndrome-related patients.

text: BACKGROUND Singapore reported its first case of Severe Acute Respiratory Syndrome (SARS) in early March 2003 and was placed on the World Health Organization's list of SARS-affected countries on March 15, 2003. During the outbreak, Tan Tock Seng Hospital was designated as the national SARS hospital in Singapore to manage all known SARS patients. Stringent infection control measures were introduced to protect healthcare workers and control intrahospital transmission of SARS. Work-flow processes for surgery were extensively modified. METHODS The authors describe the development of infection control measures, the conduct of surgical procedures, and the management of high-risk procedures during the SARS outbreak. RESULTS Forty-one operative procedures, including 15 high-risk procedures (surgical tracheostomy), were performed on SARS-related patients. One hundred twenty-four healthcare workers had direct contact with SARS patients during these procedures. There was no transmission of SARS within the operating room complex. CONCLUSIONS Staff personal protection, patient risk categorization, and reorganization of operating room workflow processes formed the key elements for the containment of SARS transmission. Lessons learned during this outbreak will help in the planning and execution of infection control measures, should another outbreak occur.


title: Crossing the species barrier: the threat of an avian influenza pandemic.

text: Avian influenza (H5N1) has recently been recognized as a new emerging infectious disease that may pose a threat to international public health. Most recent developments lead to the belief that H5N1 could become the cause of the next influenza pandemic. This review discusses the characteristics of H5N1 avian influenza virus as an emerging infectious disease with the potential for pandemic development. In addition, the current pandemic influenza alert status and guidelines for pandemic preparedness, treatment, and prevention are discussed.


title: Coronavirus JHM: a virion-associated protein kinase.

text: Coronavirus JHM contains six major proteins, one of which, the 60 000 mol. wt. nucleocapsid protein pp60, is phosphorylated. In JHM-infected cells ip 60K, the intracellular precursor to pp60 is also phosphorylated. Associated with purified JHM virions is a protein kinase which will phosphorylate pp60 and a variety of exogenous substances in vitro. The enzyme has the characteristics of a cyclic nucleotide-independent protein kinase. Both the in vivo reaction and the enzyme activity in vitro transferred the gamma-phosphate of ATP to serine residues on the nucleocapsid protein.


title: A review of evidence for immunosuppression due to porcine reproductive and respiratory syndrome virus.

text: Accounts of field disease and experimental studies involving porcine reproductive and respiratory syndrome (PRRS) are reviewed for evidence of immunomodulation or immunosuppression by the causative virus. The conclusion is that immunomodulation through infection of alveolar macrophages is likely to occur, but that it is transient and at a local level, in the lung. There is some evidence for more subtle effects via more disseminated replication or induction of apoptosis with some isolates, but more definitive studies are needed. There is some emerging evidence of interaction between PRRSV and different cells of the immune system, but its significance for the course of disease or pig health are unclear. Likewise, the current experimental evidence for any interaction of PRRSV with other pathogens is ambiguous and therefore no firm conclusions can yet be drawn. Strains of PRRSV do vary in pathogenicity, which may be related to their degree of ability to cause overt respiratory disease in the absence of other agents. Experimentally, varying degrees of interstitial pneumonia are a common histological finding. There is, as yet, no firm evidence of general immunosuppression--in fact, some contrary evidence exists in the form of observations of a transient enhancement of humoral response, possibly through polyclonal B cell activation. The basis of pathogenicity of PRRSV and of any interaction with other agents is still unknown and is likely to remain unclear. Virus interaction with the pig's immune system must be addressed before any assessment of virulence of any known or emergent strains of PRRSV can be made.


docid: gsjsdkkl

title: Wireless tissue palpation for intraoperative detection of lumps in the soft tissue.

text: In an open surgery, identification of precise margins for curative tissue resection is performed by manual palpation. This is not the case for minimally invasive and robotic procedures, where tactile feedback is either distorted or not available. In this paper, we introduce the concept of intraoperative wireless tissue palpation. The wireless palpation probe (WPP) is a cylindrical device (15 mm in diameter, 60 mm in length) that can be deployed through a trocar incision and directly controlled by the surgeon to create a volumetric stiffness distribution map of the region of interest. This map can then be used to guide the tissue resection to minimize healthy tissue loss. The wireless operation prevents the need for a dedicated port and reduces the chance of instrument clashing in the operating field. The WPP is able to measure in real time the indentation pressure with a sensitivity of 34 Pa, the indentation depth with an accuracy of 0.68 mm, and the probe position with a maximum error of 11.3 mm in a tridimensional workspace. The WPP was assessed on the benchtop in detecting the local stiffness of two different silicone tissue simulators (elastic modulus ranging from 45 to 220 kPa), showing a maximum relative error below 5%. Then, in vivo trials were aimed to identify an agar-gel lump injected into a porcine liver and to assess the device usability within the frame of a laparoscopic procedure. The stiffness map created intraoperatively by the WPP was compared with a map generated ex vivo by a standard uniaxial material tester, showing less than 8% local stiffness error at the site of the lump.


title: Human monoclonal antibodies to the S glycoprotein and related proteins as potential therapeutics for SARS.

text: Polyclonal antibodies have a century-old history of being effective against some viruses and, recently, monoclonal antibodies (mAbs) have also shown some clinical success. Human mAbs to the severe acute respiratory syndrome (SARS) coronavirus spike glycoprotein have been developed by several research groups at an amazing pace. These antibodies potently neutralize infectious virus in tissue cultures and animal models, and, alone or in combination with vaccines and other drugs, may have potential for the prevention and treatment of SARS.


title: The experimental production of diarrhoea in colostrum deprived axenic and gnotoxenic calves with enteropathogenic Escherichia coli, rotavirus, coronavirus and in a combined infection of rotavirus and E. coli.

text: We attempted to produce diarrhoea experimentally in the newborn calf by orally injecting 17 colostrum-deprived calves with two serotypes of Escherichia coli Ent+ K99+, a rotavirus and a coronavirus. With E. coli alone, a dose of 2 x 10(8) bacteria administered 24 hours after birth causes a mild attack of diarrhoea, whereas 1 x 10(10) bacteria leads to dehydration and death. An inoculation of rotavirus is followed by diarrhoea which always contains large quantities of rotavirus. These animals were anorectic for a time, but none was dehydrated or died. With coronavirus, there were large quantities of watery diarrhoea, which led to dehydration and death. The inoculation of rotavirus, not lethal in itself, followed by a similarly non lethal inoculation of E. coli in doses of 3 x 10(8) to 2 x 10(9) led to dehydration and death. The authors conclude that dehydration and death of the animal can caused by large doses of E. coli or coronavirus or by two non-lethal doses of rotavirus and E. coli administered one after the other.


title: Mass spectroscopic characterization of the coronavirus infectious bronchitis virus nucleoprotein and elucidation of the role of phosphorylation in RNA binding by using surface plasmon resonance.

text: Phosphorylation of the coronavirus nucleoprotein (N protein) has been predicted to play a role in RNA binding. To investigate this hypothesis, we examined the kinetics of RNA binding between nonphosphorylated and phosphorylated infectious bronchitis virus N protein with nonviral and viral RNA by surface plasmon resonance (Biacore). Mass spectroscopic analysis of N protein identified phosphorylation sites that were proximal to RNA binding domains. Kinetic analysis, by surface plasmon resonance, indicated that nonphosphorylated N protein bound with the same affinity to viral RNA as phosphorylated N protein. However, phosphorylated N protein bound to viral RNA with a higher binding affinity than nonviral RNA, suggesting that phosphorylation of N protein determined the recognition of virus RNA. The data also indicated that a known N protein binding site (involved in transcriptional regulation) consisting of a conserved core sequence present near the 5' end of the genome (in the leader sequence) functioned by promoting high association rates of N protein binding. Further analysis of the leader sequence indicated that the core element was not the only binding site for N protein and that other regions functioned to promote high-affinity binding.


title: Glycyrrhizin attenuates mucus production by inhibition of MUC5AC mRNA expression in vivo and in vitro.

text: We have previously indicated that glycyrrhizin (GL), a major component of licorice, has glucocorticoid-like anti-inflammatory effects in cultured airway epithelial cells and suggested its usefulness in the treatment of inflammatory respiratory diseases. On the other hand, mucus hypersecretion in the respiratory tract and goblet cell hyperplasia in the airway epithelium contribute to the morbidity and mortality associated with airway inflammatory diseases. This study, therefore, aimed to examine the effects of GL on airway mucus hyperproduction and define the mechanisms behind these effects. In an in vivo model, GL significantly attenuated goblet cell hyperplasia and MUC5AC mRNA expression in mice treated with lipopolysaccharide or interleukin-4. In addition, GL significantly attenuated MUC5AC protein and mRNA expression by tumor growth factor (TGF)-alpha in cultured NCI-H292 cells. GL also attenuated TGF-alpha-stimulated MUC5AC promoter activity in a luciferase reporter gene assay, but did not affect the stability of MUC5AC mRNA. Taken together, we concluded that GL has an inhibitory effect on mucus hyperproduction both in vivo and in vitro and that GL-mediated inhibition may be mediated through the inhibition of MUC5AC gene transcription.


title: Factors and determinants of disease emergence.

text: Emerging infectious diseases can be defined as infections that have newly appeared in a population or are rapidly increasing in incidence or geographic range. Many of these diseases are zoonoses, including such recent examples as avian influenza, severe acute respiratory syndrome, haemolytic uraemic syndrome (a food-borne infection caused by certain strains of Escherichia coli) and probably human immunodeficiency virus/acquired immune deficiency syndrome. Specific factors precipitating the emergence of a disease can often be identified. These include ecological, environmental or demographic factors that place people in increased contact with the natural host for a previously unfamiliar zoonotic agent or that promote the spread of the pathogen. These factors are becoming increasingly prevalent, suggesting that infections will continue to emerge and probably increase. Strategies for dealing with the problem include focusing special attention on situations that promote disease emergence, especially those in which animals and humans come into contact, and implementing effective disease surveillance and control.


title: [An epidemiological study on the index cases of severe acute respiratory syndrome occurred in different cities among Guangdong province].

text: OBJECTIVE To analyze the epidemiological features of the index cases of severe acute respiratory syndrome (SARS) occurred in different cities in Guangdong province and to trace for the source of infection. METHODS Standardized individual case inventory was adopted to conduct investigation on index cases and on persons who had close contact with index cases in Guangdong province. Data on the epidemiological characteristics, secondary cases and the links among index cases were analyzed. RESULTS Between November 16, 2002 and April 16, 2003, there had been 13 index cases of SARS including 3 cadres, 3 farmers, 2 retirees, 2 workers and 1 shop attendant, reported from 13 cities in Guangdong province. Between November 2002 and January 2003, there had been 7 cities reported to have identified index cases of SARS with 6 of them being infected in their own cities and 1 imported from Guangzhou city. All of the cases had no close contacts to similar patients but 6 of them later caused 2nd or even 3rd generation cases of SARS. Most cases hit young people (7/13) with a sex ratio of 1:0.6. The fatality rate of index cases was high (4/13). CONCLUSION No evidence showed that there was direct transmission among the index cases. Data regarding the geographical origin of those index cases led to the assumption that the infection had started in six cities of Pearl river delta region and the Hong Kong Special Administrative Region.


title: Detection of canine distemper virus in dogs by real-time RT-PCR.

text: Canine distemper virus is the etiological agent of a severe disease in dogs and many other carnivores. Clinical diagnosis of canine distemper is difficult due to the broad spectrum of signs that may be confounded with other respiratory and enteric diseases of dogs. Accordingly, a laboratory confirmation is required for suspected cases. In this study a real-time RT-PCR assay was developed for detection and quantitation of canine distemper virus. The assay exhibited high specificity as all the negative controls (no-template-controls and samples from healthy sero-negative dogs) and other canine pathogens were not misdetected. Up to 1 x 10(2) copies of RNA were detected by the TaqMan assay, thus revealing a high sensitivity. Quantitative TaqMan was validated on clinical samples, including various tissues and organs collected from dogs naturally infected by canine distemper virus. Urines, tonsil, conjunctival swabs and whole blood were found to contain high virus loads and therefore proved to be suitable targets for detection of canine distemper virus RNA.


title: Identification and analysis of gp116 and gp64 structural glycoproteins of yellow head nidovirus of Penaeus monodon shrimp.

text: Yellow head virus (YHV) is a major agent of disease in farmed penaeid shrimp. YHV virions purified from infected shrimp contain three major structural proteins of molecular mass 116 kDa (gp116), 64 kDa (gp64) and 20 kDa (p20). Two different staining methods indicated that the gp116 and gp64 proteins are glycosylated. Here we report the complete nucleotide sequence of ORF3, which encodes a polypeptide of 1666 amino acids with a calculated molecular mass of 185 713 Da (pI=6.68). Hydropathy analysis of the deduced ORF3 protein sequence identified six potential transmembrane helices and three ectodomains containing multiple sites for potential N-linked and O-linked glycosylation. N-terminal sequence analysis of mature gp116 and gp64 proteins indicated that each was derived from ORF3 by proteolytic cleavage of the polyprotein between residues Ala(228) and Thr(229), and Ala(1127) and Leu(1128), located at the C-terminal side of transmembrane helices 3 and 5, respectively. Comparison with the deduced ORF3 protein sequence of Australian gill-associated virus (GAV) indicated 83 % amino acid identity in gp64 and 71 % identity in gp116, which featured two significant sequence deletions near the N terminus. Database searches revealed no significant homology with other proteins. Recombinant gp64 expressed in E. coli with and without the C-terminal transmembrane region was shown to react with antibody raised against native gp64 purified from virions.


title: Does this patient have influenza?

text: CONTEXT Influenza vaccination lowers, but does not eliminate, the risk of influenza. Making a reliable, rapid clinical diagnosis is essential to appropriate patient management that may be especially important during shortages of antiviral agents caused by high demand. OBJECTIVES To systematically review the precision and accuracy of symptoms and signs of influenza. A secondary objective was to review the operating characteristics of rapid diagnostic tests for influenza (results available in <30 min). DATA SOURCES Structured search strategy using MEDLINE (January 1966-September 2004) and subsequent searches of bibliographies of retrieved articles to identify articles describing primary studies dealing with the diagnosis of influenza based on clinical signs and symptoms. The MEDLINE search used the Medical Subject Headings EXP influenza or EXP influenza A virus or EXP influenza A virus human or EXP influenza B virus and the Medical Subject Headings or terms EXP sensitivity and specificity or EXP medical history taking or EXP physical examination or EXP reproducibility of results or EXP observer variation or symptoms.mp or clinical signs.mp or sensitivity.mp or specificity.mp. STUDY SELECTION Of 915 identified articles on clinical assessment of influenza-related illness, 17 contained data on the operating characteristics of symptoms and signs using an independent criterion standard. Of these, 11 were eliminated based on 4 inclusion criteria and availability of nonduplicative primary data. DATA EXTRACTION Two authors independently reviewed and abstracted data for estimating the likelihood ratios (LRs) of clinical diagnostic findings. Differences were resolved by discussion and consensus. DATA SYNTHESIS No symptom or sign had a summary LR greater than 2 in studies that enrolled patients without regard to age. For decreasing the likelihood of influenza, the absence of fever (LR, 0.40; 95% confidence interval [CI], 0.25-0.66), cough (LR, 0.42; 95% CI, 0.31-0.57), or nasal congestion (LR, 0.49; 95% CI, 0.42-0.59) were the only findings that had summary LRs less than 0.5. In studies limited to patients aged 60 years or older, the combination of fever, cough, and acute onset (LR, 5.4; 95% CI, 3.8-7.7), fever and cough (LR, 5.0; 95% CI, 3.5-6.9), fever alone (LR, 3.8; 95% CI, 2.8-5.0), malaise (LR, 2.6; 95% CI, 2.2-3.1), and chills (LR, 2.6; 95% CI, 2.0-3.2) increased the likelihood of influenza to the greatest degree. The presence of sneezing among older patients made influenza less likely (LR, 0.47; 95% CI, 0.24-0.92). CONCLUSIONS Clinical findings identify patients with influenza-like illness but are not particularly useful for confirming or excluding the diagnosis of influenza. Clinicians should use timely epidemiologic data to ascertain if influenza is circulating in their communities, then either treat patients with influenza-like illness empirically or obtain a rapid influenza test to assist with management decisions.


title: Impact of zoonoses on human health.

text: By reviewing the most significant zoonotic disease outbreaks that have occurred mostly during the past ten years, the author provides a clear idea of how varied these diseases can be in regard to their aetiological agent, size and direct impact on public health. Most examples involve emerging zoonotic diseases caused by viruses and prions and transmitted to humans by a bite, close contact with affected live animals or carcasses, or through the consumption of their tissues. These outbreaks vary from very small and localised clusters of individual cases to millions of deaths, as reported during the past influenza pandemics. The author also shows that even for the larger outbreaks, the direct impact on public health measured by the morbidity and mortality of zoonoses is largely inferior to that of major communicable diseases that affect only humans, particularly human tuberculosis, malaria, HIV/AIDS. However, it is very difficult to predict the outcome on public health of these emerging zoonotic diseases since transmission patterns are not always sufficiently understood to assess this impact accurately. In addition, new modes of agent transmission may compound the initial impact on public health. Finally, the author indicates additional reasons that explain why these diseases are important by placing special emphasis on the financial losses recorded in both human and animal health and also the societal non-monetary losses these diseases can incur. Lessons learnt following major crises generated by the emergence of zoonotic diseases, such as bovine spongiform encephalopathy, severe acute respiratory syndrome and avian influenza, are provided.


title: Parietex™ Composite mesh versus DynaMesh®-IPOM for laparoscopic incisional and ventral hernia repair: a retrospective cohort study.

text: INTRODUCTION Laparoscopic incisional and ventral hernia repair (LIVHR) is widely accepted and safe but the type of mesh used is still debated. We retrospectively compared postoperative outcomes with two different meshes commonly used in LIVHR. METHODS This is a retrospective study of patients who underwent incisional hernia repair between January 2008 and December 2010. Two meshes were used: Parietex™ Composite (Covidien, New Haven, CT, USA) and the DynaMesh®-IPOM (FEG Textiltechnik mbH, Aachen, Germany). The two groups were compared with respect to recurrence rates, incidence of seroma and intestinal obstruction. RESULTS Among the 88 patients who underwent LIVHR, 75 patients (85.2%) presented with primary incisional hernia, 10 (11.4%) presented with a first recurrence and 3 (3.4%) presented with a second recurrence. Median follow-up was 53.6 months (range 40-61 months). 12.9% of patients had recurrence in the Parietex™ Composite mesh group (n=62) in comparison to 3.8% in the DynaMesh®-IPOM mesh group (n=26; P=0.20). DynaMesh®-IPOM was associated with a significantly higher incidence of intestinal obstruction secondary to adhesions (11.5% vs. 0%, P=0.006) and lower incidence of seroma and haematoma formation compared to Parietex™ composite mesh group (0% vs. 6.4% of patients; P=0.185). CONCLUSIONS LIVHR is a safe and feasible technique. Dynamesh®-IPOM is associated with a significantly higher incidence of adhesion related bowel obstruction, albeit with a lower incidence of recurrence, seroma and haematoma formation compared with Parietex™ Composite mesh. However, there is a need for further well-designed, multicentre randomised controlled studies to investigate the use of these meshes.


title: Lessons from the epidemiological surveillance program, during the influenza A (H1N1) virus epidemic, in a reference university hospital of Southeastern Brazil.

text: INTRODUCTION The case definition of influenza-like illness (ILI) is a powerful epidemiological tool during influenza epidemics. METHODS A prospective cohort study was conducted to evaluate the impact of two definitions used as epidemiological tools, in adults and children, during the influenza A H1N1 epidemic. Patients were included if they had upper respiratory samples tested for influenza by real-time reverse transcriptase polymerase chain reaction during two periods, using the ILI definition (coughing + temperature ≤ 38ºC) in period 1, and the definition of severe acute respiratory infection (ARS) (coughing + temperature ≤ 38ºC and dyspnoea) in period 2. RESULTS The study included 366 adults and 147 children, covering 243 cases of ILI and 270 cases of ARS. Laboratory confirmed cases of influenza were higher in adults (50%) than in children (21.6%) ( p < 0.0001) and influenza infection was more prevalent in the ILI definition (53%) than ARS (24.4%) (p < 0.0001). Adults reported more chills and myalgia than children (p = 0.0001). Oseltamivir was administered in 58% and 46% of adults and children with influenza A H1N1, respectively. The influenza A H1N1 case fatality rate was 7% in adults and 8.3% in children. The mean time from onset of illness until antiviral administration was 4 days. CONCLUSIONS The modification of ILI to ARS definition resulted in less accuracy in influenza diagnosis and did not improve the appropriate time and use of antiviral medication.


title: Long term evaluation of radiographic disease progression in a subset of patients with rheumatoid arthritis treated with leflunomide beyond 2 years.

text: OBJECTIVES To assess the effect of long term (>2 years) leflunomide treatment on radiographic progression in patients with RA. METHODS Patients treated with leflunomide for >2 years in one of three phase III trials and subsequent extensions, for whom paired, evaluable radiographs at baseline and study end point were available, were included. Radiographs of hands and feet were assessed according to the modified Sharp/van der Heijde scoring method, for erosion, joint space narrowing, and total score. Changes from baseline were assessed, and a predicted yearly progression rate estimated for each patient. RESULTS 128 of the original 824 patients were included, with mean disease duration 5.1 years and mean leflunomide treatment duration 4.3 years until the final x ray examination. The mean change from baseline in total score was 8.6 with yearly adjusted rate 1.9, and the median change was 2 with yearly adjusted rate 0.5, compared with 7.9 and 4.9, respectively, before leflunomide treatment. After treatment, the rate improved in 92/128 (72%) patients and deteriorated in 21/128 (16%). In 42 (33%) patients who had a total score >0 at baseline, no radiographic progression occurred after leflunomide treatment. CONCLUSIONS In a subset of patients who continued treatment long term, leflunomide treatment reduced the rate of radiographic damage.


title: Feasibility Assessment of a New Surveillance Tool for Respiratory Protective Devices Used in U.S. Healthcare.

text: Background Respiratory protective devices (RPDs) are used for infection prevention in healthcare settings during routine patient care and public health emergencies. In recent years, healthcare systems have experienced shortages of RPDs during outbreaks of infectious diseases, in part due to a lack of information about their availability. New tools to track RPD inventories may improve accessibility during an emergency. Investigators at Vanderbilt University have identified four major themes that influence RPD use for infection prevention: hospital preparedness, responsiveness to airborne pathogens, potential exposure outcomes, and infection control practices related to respirator effectiveness. Based on these findings, an RPD surveillance tool (RST) was developed to collect and share near real-time data about RPD supplies in healthcare facilities. The objective of this study was to conduct a feasibility assessment of this RST. Methods The new online surveillance tool was implemented at four large, urban, acute care U.S. hospitals in January 2014; data was collected about RPD inventory, tracking systems, hospital characteristics, and utility of gathered information. Results The RST was implemented successfully and without difficulty at hospitals that had 78 to 90 percent occupancy rates. Participating hospitals reported that the RST (1) provided value for benchmarking their RPD supply, (2) promoted understanding about RPD accessibility among hospital systems engaged in infection control, and (3) served as a means to assess RPD program quality. Conclusion Implementation of this newly developed RST is feasible and appears to have utility in U.S. hospitals for tracking and understanding RPD use for routine healthcare delivery and public health emergencies.


title: Progress toward global eradication of dracunculiasis.

text: The plan for the global eradication of dracunculiasis (i.e., Guinea worm disease) was developed in October 1980, when dracunculiasis was known to be endemic in 16 African countries, India, and Pakistan. In 1991, the World Health Assembly adopted a resolution to eradicate dracunculiasis by the end of 1995. This report summarizes the substantial progress toward eradication of dracunculiasis since the beginning of this effort.


title: Recent advances of microfluidics in Mainland China.

text: The history and current status of research on microfluidics in China is summarized in this review. The recent representative contributions in this field by Chinese scientists are cited. A perspective on some trends in future development of this field in China is presented.


title: Acute viral infections of upper respiratory tract in elderly people living in the community: comparative, prospective, population based study of disease burden.

text: OBJECTIVE To evaluate the disease burden of upper respiratory infections in elderly people living at home. DESIGN Prospective surveillance of elderly people. INTERVENTION None. SETTING Leicestershire, England SUBJECTS 533 subjects 60 to 90 years of age. MAIN OUTCOME MEASURES Pathogens, symptoms, restriction of activity, duration of illness, medical consultations, interval between onset of illness and medical consultation, antibiotic use, admission to hospital, and death. RESULTS 231 pathogens were identified for 211 (43%) of 497 episodes for which diagnostic specimens were available: 121 (52%) were rhinoviruses, 59 (26%) were coronaviruses, 22 (9.5%) were influenza A or B, 17 (7%) were respiratory syncytial virus, 7 (3%) were parainfluenza viruses, and 3 (1%) were Chlamydia species; an adenovirus and Mycoplasma pneumoniae caused one infection each. Infections occurred at a rate of 1.2 episodes per person per annum (95% confidence interval 1.0 to 1.7; range 0-10) and were clinically indistinguishable. Lower respiratory tract symptoms complicated 65% of upper respiratory infections and increased the medical consultation rate 2.4-fold (chi 2 test P < 0.001). The median interval between onset of illness and medical consultation was 3 days for influenza and 5 days for other infections. Rhinoviruses caused the greatest disease burden overall followed by episodes of unknown aetiology, coronaviruses, influenza A and B, and respiratory syncytial virus. CONCLUSIONS Respiratory viruses cause substantial morbidity in elderly people. Although respiratory syncytial virus and influenza cause considerable individual morbidity, the burden of disease from rhinovirus infections and infections of unknown aetiology seems greater overall. The interval between onset of illness and consultation together with diagnostic difficulties raises concern regarding the role of antiviral drugs in treating influenza.


AmBisome administration for Candida albicans shunt infections.


Zoonotic potential of emerging animal diseases.


The structure of the canine coronavirus.


title: Preclinical evaluation of 68Ga-DOTA-minigastrin for the detection of cholecystokinin-2/gastrin receptor-positive tumors.

text: In comparison to somatostatin receptor scintigraphy, gastrin receptor scintigraphy using 111In-DTPA-minigastrin (MG0) showed added value in diagnosing neuroendocrine tumors. We investigated whether the 68Ga-labeled gastrin analogue DOTA-MG0 is suited for positron emission tomography (PET), which could improve image quality. Targeting of cholecystokinin-2 (CCK2)/gastrin receptor-positive tumor cells with DOTA-MG0 labeled with either 111In or 68Ga in vitro was investigated using the AR42J rat tumor cell line. Biodistribution was examined in BALB/c nude mice with a subcutaneous AR42J tumor. In vivo PET imaging was performed using a preclinical PET-computed tomographic scanner. DOTA-MG0 showed high receptor affinity in vitro. Biodistribution studies revealed high tumor uptake of 68Ga-DOTA-MG0: 4.4 ± 1.3 %ID/g at 1 hour postinjection. Coadministration of an excess unlabeled peptide blocked the tumor uptake (0.7 ± 0.1 %ID/g), indicating CCK2/gastrin receptor-mediated uptake (p = .0005). The biodistribution of 68Ga-DOTA-MG0 was similar to that of 111In-DOTA-MG0. Subcutaneous and intraperitoneal tumors were clearly visualized by small-animal PET imaging with 5 MBq 68Ga-DOTA-MG0. 111In- and 68Ga-labeled DOTA-MG0 specifically accumulate in CCK2/gastrin receptor-positive AR42J tumors with similar biodistribution apart from the kidneys. AR42J tumors were clearly visualized by microPET. Therefore, 68Ga-DOTA-MG0 is a promising tracer for PET imaging of CCK2/gastrin receptor-positive tumors in humans.


title: [A three-year follow-up study on sera specific antibody in severe acute respiratory syndrome cases after the onset of illness].

text: OBJECTIVE To study the variation of specific antibody among convalescent of severe acute respiratory syndrome (SARS) patients through a three-year program. METHODS Sera samples were collected from SARS cases in the 5th, 20th and 35th month after onset of the illness. The SARS-CoV specific antibody was detected for all of them by ELISA and neutralized test simultaneously. The titer of neutralizing antibodies was calculated using Reed-Muench method, and the comparison between different time groups was analyzed regarding the variance of data on repeated measures after logarithm conversion. RESULTS 13, 17 and 13 sera samples were collected in the 5th, 20th and 35th month after onset. Results showed that despite the fact that the positive rates of ELISA antibody were 100%, 82.4% and 84.6% respectively,the neutralizing antibody was still positive for all the samples. The average neutralizing antibody titers were 1:43 (1:16-1:203), 1:36 (1:17-1:59) and 1:21 (1:10-1:39) on the 5th, 20th and 35th month after onset, and the differences were statistically significant (F = 60.419, P < 0.001). On the 35th month after the onset, 30.8% (4/13) of the patients were still having the neutralizing antibody level of above 1:36, but the neutralizing antibody level in another 30.8% (4/13) of the patients had decreased to as low as 1:10, when the cut-off level was set as 1:8. CONCLUSION Results of the study indicated that the neutralizing antibody of SARS cases could last for at least three years, but the sera specific antibody in SARS cases decreased gradually when time went by. However, neutralizing antibody in some of the cases decreased to a lower level on the 35th month. Further follow-up study was worthwhile to observe the long-lasting profile of antibody existence on SARS cases.


title: Molecular biology of umbraviruses: phantom warriors.

text: The genomes of umbraviruses differ from those of most other viruses in that they do not encode a coat protein, and thus no virus particles are formed in infected plants. Protection of umbraviral RNA outside the host plant, during vector transmission, utilizes the coat protein of an assistor luteovirus, but this review focuses on the mechanisms that compensate for the lack of a coat protein in processes within the host plant. As well as an RNA-dependent RNA polymerase, umbravirus genomes encode two other proteins from almost completely overlapping open reading frames. One of these is a cell-to-cell movement protein that can mediate the transport of homologous and heterologous viral RNAs through plasmodesmata without the participation of a coat protein. The other, the ORF3 protein, binds to viral RNA to form filamentous ribonucleoprotein particles that have elements of helical structure. It serves to stabilize the RNA and facilitates its transport through the vascular system of the plant. It may also be involved in protection of the viral RNA from the plant's defensive RNA-silencing response, although it is not a suppressor of silencing. The ORF3 protein also enters the cell nucleus, specifically targeting the nucleolus. Although the function of this localization is unknown, the ORF3 protein may provide a valuable tool for investigating plant nucleolar function.


title: Sialic acid binding activity of transmissible gastroenteritis coronavirus affects sedimentation behavior of virions and solubilized glycoproteins.

text: The sedimentation behavior of transmissible gastroenteritis coronavirus (TGEV) was analyzed. Upon sucrose gradient centrifugation, the major virus band was found at a density of 1.20 to 1.22 g/cm(3). This high density was observed only when TGEV with a functional sialic acid binding activity was analyzed. Mutants of TGEV that lacked sialic acid binding activity due to a point mutation in the sialic acid binding site of the S protein were mainly recovered at a lower-density position on the sucrose gradient (1.18 to 1.19 g/cm(3)). Neuraminidase treatment of purified virions resulted in a shift of the sedimentation value from the higher to the lower density. These results suggest that binding of sialoglycoproteins to the virion surface is responsible for the sedimentation behavior of TGEV. When purified virions were treated with octylglucoside to solubilize viral glycoproteins, ultracentrifugation resulted in sedimentation of the S protein of TGEV. However, when neuraminidase-treated virions or mutants with a defective sialic acid binding activity were analyzed, the S protein remained in the supernatant rather than in the pellet fraction. These results indicate that the interaction of the surface protein S with sialoglycoconjugates is maintained after solubilization of this viral glycoprotein by detergent treatment.


title: Mapping of the coronavirus membrane protein domains involved in interaction with the spike protein.

text: The coronavirus membrane (M) protein is the key player in virion assembly. One of its functions is to mediate the incorporation of the spikes into the viral envelope. Heterotypic interactions between M and the spike (S) protein can be demonstrated by coimmunoprecipitation and by immunofluorescence colocalization, after coexpression of their genes in eukaryotic cells. Using these assays in a mutagenetic approach, we have mapped the domains in the M protein that are involved in complex formation between M and S. It appeared that the 25-residue luminally exposed amino-terminal domain of the M protein is not important for M-S interaction. A 15-residue deletion, the insertion of a His tag, and replacement of the ectodomain by that of another coronavirus M protein did not affect the ability of the M protein to associate with the S protein. However, complex formation was sensitive to changes in the transmembrane domains of this triple-spanning protein. Deletion of either the first two or the last two transmembrane domains, known not to affect the topology of the protein, led to a considerable decrease in complex formation, but association was not completely abrogated. Various effects of changes in the part of the M protein that is located at the cytoplasmic face of the membrane were observed. Deletions of the extreme carboxy-terminal tail appeared not to interfere with M-S complex formation. However, deletions in the amphipathic domain severely affected M-S interaction. Interestingly, changes in the amino-terminal and extreme carboxy-terminal domains of M, which did not disrupt the interaction with S, are known to be fatal to the ability of the protein to engage in virus particle formation (C. A. M. de Haan, L. Kuo, P. S. Masters, H. Vennema, and P. J. M. Rottier, J. Virol. 72:6838-6850, 1998). Apparently, the structural requirements of the M protein for virus particle assembly differ from the requirements for the formation of M-S complexes.


title: Renal cell carcinoma and COVID-19 pandemia: Management strategies adapted to current practice.

text: The COVID-19 pandemic caused by SARS-CoV-2 virus has caused an important health impact that has affected renal cell carcinoma management, among other urology areas. The high cancellation rate of surgeries, including those related to renal cancer, will cause an inevitable healthcare overload and probably a potential negative impact on its oncological outcomes, especially in locally advanced and metastatic renal cancer. Kidney cancer scenarios are quite different depending on their stage, distinguishing mainly between low priority of localized disease or high priority of locally advanced and metastatic under active treatment. The unknown pandemic duration and possibly fluctuating prevalence of the virus are likely to force an adaptation in the management of renal cell carcinoma among urology and oncology departments, ideally individualized ona case-by-case basis within multidisciplinary units. To this end, we present algorithms and tables regarding renal cell carcinoma management adapted to the COVID-19 period and stratified according to oncological stage, which might be useful for specialists dedicated to this uro-oncology area.


title: Helminthic eosinophilic meningitis: emerging zoonotic diseases in the South.

text: Today most emerging infectious diseases, such as West Nile virus and severe acute respiratory syndrome (SARS), arise in the natural environment as zoonoses and are often imported into the United States (US). The most common helminthic infections that can cause eosinophilic meningitis (EoM) in the US, neuroangiostrongyliasis and baylisascariasis, share many of the characteristics of emerging infectious diseases. Neuroangiostrongyliasis, a rodent zoonosis caused by the rat lungworm, Angiostrongylus cantonensis, is now endemic in the US following the importation of infected rats on container ships and African land snails, the parasite's intermediate hosts, as biological controls and exotic pets. Baylisascariasis, a raccoon zoonosis, caused by the raccoon roundworm, Baylisascaris procyonis, has extended its US distribution range from suburban neighborhoods in the northern US to the Southeast and West Coast since the 1980s. Both A. cantonensis and B. procyonis are now enzootic in Louisiana and have caused EoM in humans. This review analyzes scientific articles selected by MEDLINE search, 1966-2008, in order to assess the evolving epidemiology of EoM in the US, and specifically in Louisiana; and to alert Louisiana clinicians to populations at increased risk of helminthic EoM as a result of age, ethnicity, lifestyle, food choices, location of permanent residence, or recent travel in the Americas or Caribbean. Most parasitic diseases causing EoM are no longer confined to tropical countries; they are now endemic in the US and in Louisiana and more cases may be anticipated.


title: Unintentional poisoning deaths--United States, 1999-2004.

text: In 2004, poisoning was second only to motor-vehicle crashes as a cause of death from unintentional injury in the United States . Nearly all poisoning deaths in the United States are attributed to drugs, and most drug poisonings result from the abuse of prescription and illegal drugs. Previous reports have indicated a substantial increase in unintentional poisoning mortality during the 1980s and 1990s. To further examine this trend, CDC analyzed the most current data from the National Vital Statistics System. This report summarizes the results of that analysis, which determined that poisoning mortality rates in the United States increased each year from 1999 to 2004, rising 62.5% during the 5-year period. The largest increases were among females (103.0%), whites (75.8%), persons living in the southern United States (113.6%), and persons aged 15-24 years (113.3%). Larger rate increases occurred in states with mostly rural populations. Rates for drug poisoning deaths increased 68.3%, and mortality rates for poisonings by other substances increased 1.3%. The largest increases were in the "other and unspecified," psychotherapeutic, and narcotic drug categories. The results suggest that more aggressive regulatory, educational, and treatment measures are necessary to address the increase in fatal drug overdoses.


title: Simultaneous detection of North American and European porcine reproductive and respiratory syndrome virus using real-time quantitative reverse transcriptase-PCR.

text: Porcine reproductive and respiratory syndrome (PRRS) is 1 of the most economically important diseases of swine. Detection of the etiologic agent, PRRS virus (PRRSV), represents a diagnostic challenge due to the heterogeneity of field isolates as well as the propensity for swine to develop persistent infection in which virus is difficult to detect. Recently European (EU) lineage PRRSV isolates, which are genetically divergent from North American (NA) isolates, have been introduced into NA swine further complicating efforts to diagnose this disease. In this study, real-time (TaqMan) reverse transcriptase (RT)-PCR assays were developed for multiplex detection, differentiation, and quantification of NA and EU PRRSV field isolates. Oligonucleotide primers and dual-labeled probes were selected from conserved regions of open-reading frame 7 and the 3'-untranslated region. The real-time RT-PCR assays described for the NA or EU genotype of PRRSV detected viral RNA from 83/83 strains (74 NA; 9 EU) previously isolated by cell culture between 1992 and 2003. The analytical sensitivity of both assays was consistently found to be less than a single TCID50, which corresponded to 5-10 RNA molecules, and was not significantly reduced when the reactions were performed in a multiplex format. When performing multiplex reactions, sensitive detection was possible even when 1 viral RNA concentration was up to 5,000-fold higher than the second. The diagnostic sensitivity and specificity of the multiplex reaction was found to be at a minimum equivalent to that of both nested RT-PCR and virus isolation.


title: Role of the mitochondrial signaling pathway in murine coronavirus-induced oligodendrocyte apoptosis.

text: A previous study demonstrated that infection of rat oligodendrocytes by mouse hepatitis virus (MHV) resulted in apoptosis, which is caspase dependent (Y. Liu, Y. Cai, and X. Zhang, J. Virol. 77:11952-11963, 2003). Here we determined the involvement of the mitochondrial pathway in MHV-induced oligodendrocyte apoptosis. We found that caspase-9 activity was 12-fold higher in virus-infected cells than in mock-infected cells at 24 h postinfection (p.i.). Pretreatment of cells with a caspase-9 inhibitor completely blocked caspase-9 activation and partially inhibited the apoptosis mediated by MHV infection. Analyses of cytochrome c release further revealed an activation of the mitochondrial apoptotic pathway. Stable overexpression of the two antiapoptotic proteins Bcl-2 and Bcl-xL significantly, though only partially, blocked apoptosis, suggesting that activation of the mitochondrial pathway is partially responsible for the apoptosis. To identify upstream signals, we determined caspase-8 activity, cleavage of Bid, and expression of Bax and Bad by Western blotting. We found a drastic increase in caspase-8 activity and cleavage of Bid at 24 h p.i. in virus-infected cells, suggesting that Bid may serve as a messenger to relay the signals from caspase-8 to mitochondria. However, treatment with a caspase-8 inhibitor only slightly blocked cytochrome c release from the mitochondria. Furthermore, we found that Bax but not Bad was significantly increased at 12 h p.i. in cells infected with both live and UV-inactivated viruses and that Bax activation was partially blocked by treatment with the caspase-8 inhibitor. These results thus establish the involvement of the mitochondrial pathway in MHV-induced oligodendrocyte apoptosis.


title: [Pregnant women complicated with COVID-19: a clinical analysis of 3 cases].

text: OBJECTIVE To analyze the clinical characteristics and pregnancy outcomes of pregnant women complicated with coronavirus disease 2019 (COVID-19). METHODS The clinical data of 3 pregnant women with COVID-19 admitted to the First Affiliated Hospital of Zhejiang University School of Medicine from January 19 to February 10, 2020 were retrospectively analyzed. RESULTS There was one case in the first-trimester pregnancy (case 1), one in the second-trimester pregnancy (case 2) and one in third-trimester pregnancy (case 3). Cough, fever, fatigue, lung imaging changes were the main manifestations. The white cell count, lymphocyte percentage had no significantly changes in case 1 and case 3, while the levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), IL-6 and IL-10 elevated. The lymphocyte count and lymphocyte percentage decreased and the inflammatory indicators significantly increased in case 2. All patients were treated with antiviral, antitussive, oxygen inhalation; case 3 received glucocorticoids, case 2 with severe illness received glucocorticoids and additionally gamma globulin. All three cases were cured and discharged. Case 1 with early pregnancy chose to terminate pregnancy after discharge; case 2 chose to continue pregnancy without obstetric complications; and case 3 had cesarean section delivery due to abnormal fetal heart monitoring. CONCLUSIONS The report shows that COVID-19 in pregnancy women could be cured with active treatment, and the maternal and fetal outcomes can be satisfactory.


title: Analysis of a recombinant mouse hepatitis virus expressing a foreign gene reveals a novel aspect of coronavirus transcription.

text: We have inserted heterologous genetic material into the nonessential gene 4 of the coronavirus mouse hepatitis virus (MHV) in order to test the applicability of targeted RNA recombination for site-directed mutagenesis of the MHV genome upstream of the nucleocapsid (N) gene and to develop further genetic tools for site-directed mutagenesis of structural genes other than N. Initially, a 19-nucleotide tag was inserted into the start of gene 4a of MHV strain A59 with the N gene deletion mutant Alb4 as the recipient virus. In further work, the entire gene for the green fluorescent protein (GFP) was inserted in place of gene 4, creating the currently largest known RNA virus. The expression of GFP was demonstrated by Western blot analysis of infected cell lysates; however, the level of GFP expression was not sufficient to allow detection of fluorescence of viral plaques. Northern blot analysis of transcripts of GFP recombinants showed the expected alteration of the pattern of the nested MHV subgenomic mRNAs. Surprisingly, though, GFP recombinants also produced an RNA species that was the same size as wild-type mRNA4. Analysis of the 5' end of this species revealed that it was actually a collection of mRNAs originating from 10 different genomic fusion sites, none possessing a canonical intergenic sequence. The finding of these aberrant mRNAs suggests that long-range RNA or the ribonucleoprotein structure of the MHV genome can sometimes be the sole determinant of the site of initiation of transcription.


title: Local health department and academic partnerships: education beyond the ivy walls.

text: The National Association of County and City Health Officials (NACCHO) is the national organization representing local health departments. NACCHO supports efforts that protect and improve the health of all people and all communities by promoting national policy, developing resources and programs, seeking health equity, and supporting effective local public health practice and systems.


title: Bacteria and viruses in maxillary sinuses of patients with primary hypogammaglobulinemia.

text: OBJECTIVE To study bacteria and viruses in maxillary sinuses of patients with primary hypogammaglobulinemia receiving immunoglobulin therapy. DESIGN Prospective cross-sectional study during 6 months. SETTING Tertiary care university hospital. PATIENTS Seventeen patients with primary hypogammaglobulinemia (10 males and 7 females; mean age, 39 years [age range, 11-71 years]). Sixteen patients had common variable immunodeficiency, and 1 patient had X-linked agammaglobulinemia. MAIN OUTCOME MEASURES Magnetic resonance imaging and x-ray imaging of paranasal sinuses when patients did not have signs of acute infection and reevaluation 6 months later. Maxillary sinus aspiration and lavage were performed at a follow-up visit. Sinus fluid analysis for bacteria and viruses was performed by culture and by polymerase chain reaction. A questionnaire on symptoms related to sinusitis was administered during the follow-up period. RESULTS Among 17 patients, 9 (53%) had radiologically defined sinusitis without subjective symptoms at study enrollment. At reevaluation 6 months later, radiological findings remained unchanged in two thirds of the patients. Among 15 patients, bacteria were found in sinus lavage samples from 13 patients, and viruses were found in samples from 7 patients. Eight patients had 2 pathogens or more on bacterial culture. Rhinovirus was identified from sinus lavage samples in 5 patients (33%), enterovirus in 3 patients (20%), and respiratory syncytial virus in 1 patient (7%). Pathogenic bacteria were found in maxillary sinuses of all patients who tested positive for rhinovirus and enterovirus. No fungi were found. During the follow-up period, 6 patients reported mucopurulent drainage. CONCLUSIONS Bacteria and viruses were commonly found in maxillary sinuses of patients with primary hypogammaglobulinemia. Yearly evaluation by an ear, nose, and throat surgeon is recommended.


title: Ultrastructure of the replication site in Taura syndrome virus (TSV)-infected cells.

text: Taura syndrome virus (TSV) is a member of the family Dicistroviridae that infects Pacific white shrimp Litopenaeus vannamei (also called Penaeus vannamei), and its replication strategy is largely unknown. To identify the viral replication site within infected shrimp cells, the viral RNA was located in correlation with virus-induced membrane rearrangement. Ultrastructural changes in the infected cells, analyzed by transmission electron microscopy (TEM), included the induction and proliferation of intracellular vesicle-like membranes, while the intracytoplasmic inclusion bodies and pyknotic nuclei indicative of TSV infection were frequently seen. TSV plus-strand RNA, localized by electron microscopic in situ hybridization (EM-ISH) using TSV-specific cDNA probes, was found to be associated with the membranous structures. Moreover, TSV particles were observed in infected cells by TEM, and following EM-ISH, they were also seen in close association with the proliferating membranes. Taken together, our results suggest that the membranous vesicle-like structures carry the TSV RNA replication complex and that they are the site of nascent viral RNA synthesis. Further investigations on cellular origins and biochemical compositions of these membranous structures will elucidate the morphogenesis and propagation strategy of TSV.


title: Inhibition of coronavirus 229E replication by actinomycin D.

text: The yields of human coronavirus 229E grown in L132 cells were markedly inhibited by actinomycin D, the 50% inhibitory dose being 0.1 micron/ml. Inhibition was maximal during the early phase of virus replication, did not appear to involve viral RNA synthesis per se, and was shown to be dependent on the input multiplicity of infection.


Detection of rotavirus in fecal samples from calves by a cell culture indirect immunofluorescence, an Ag-capture ELISA, a tissue culture ELISA, and a commercial Ag-capture ELISA.


Response by Menon et al to Letter Regarding Article, "Analysis of Workflow and Time to Treatment on Thrombectomy Outcome in the Endovascular Treatment for Small Core and Proximal Occlusion Ischemic Stroke (ESCAPE) Randomized, Controlled Trial".


Severe osteochondrosis in two 10-month-old beef calves.


title: [Acute respiratory failure and noninvasive positive pressure ventilation treatment in patients with severe acute respiratory syndrome].

text: OBJECTIVE To describe the blood gas features and short-term outcomes of noninvasive positive pressure ventilation (NPPV) treatment for acute respiratory failure (ARF) in patients with severe acute respiratory syndrome (SARS) in this retrospective case series study sought. METHODS Between April 22 and May 1, 2003, 120 clinically compatible SARS patients were admitted to a special hospital for infectious diseases in Beijing. All patients' records were reviewed. The outcome variables of NPPV therapy were analyzed. RESULTS At 6 weeks of SARS onset, 25% (30/120) patients had respiratory failure with ALI/ARDS. 16 (53%) had hypercapnia (PaCO(2) > 45 mm Hg) during the course of SARS. 28 of them received NPPV therapy. One was intolerable to NPPV treatment. In the remaining 27 patients, NPPV was initiated 1.2 +/- 1.6 (0 - 10) days after onset of respiratory failure. One hour of NPPV therapy led to significant increases in PaO(2), SpO(2) and PaO(2)/FiO(2), and decrease in respiratory rate (all P < 0.01). 18 of the 27 patients were weaned successfully from NPPV. The mean duration of NPPV use was 10 +/- 6 (5 - 30) days. In addition to one patient who was intolerable to NPPV treatment, intubation was required in other 9 patients who initially had a favorable response to NPPV. Remarkable pulmonary barotrauma was noted in 7 of the 120 (5.8%) patients with SARS, among which 1 developed 2 days after intubation, and 6 occurred while the patients were on NPPV, the incidence being 22% (6/27) in patients using NPPV. The fatality rate at 13 weeks of SARS onset was 6.7% (8/120) in the patient cohort, and 26.7% (8/30) in the ALI/ARDS patients. No SARS occurred in medical staff taking care of NPPV patients. CONCLUSIONS Some of the SARS patients with ALI/ARDS could experience CO(2) retention, which might be related to the impairment of respiratory muscles. It is proved to be feasible and appropriate for NPPV treatment of respiratory failure in SARS patients who were at high risk of intubation related complications. As there was a high incidence of remarkable barotrauma, a careful lung protective strategy is necessary during the administration of NPPV as well as invasive mechanical ventilation.


title: Health policy in Asia and the Pacific: Navigating local needs and global challenges.

text: Asia and the Pacific are undergoing a remarkable economic transformation which is occurring at an exceptional pace. There is clear evidence of an equally rapid epidemiological transition in the region. This paper sets out the policy challenges of building healthy societies in the context of rapid economic change. The region's location at the crossroads of contemporary globalization, resulting in intensified population mobility, large-scale trade and investment, and pressures to take collective action on shared problems, adds to the complexity of this task. The paper argues that health is integral to building stable and sustainable societies, and that there are opportunities to develop more holistic approaches that bring together hitherto separate policy spheres.


Another case of use of the ProSeal laryngeal mask airway in a difficult obstetric airway.


title: Update: severe acute respiratory syndrome--United States, June 4, 2003.

text: CDC continues to work with state and local health departments, the World Health Organization (WHO), and other partners to investigate cases of severe acute respiratory syndrome (SARS). This report updates SARS cases reported worldwide and in the United States and summarizes changes in CDC's recommendations for travel to Singapore and Hong Kong and the resulting modification to the interim U.S. case definition for SARS.


title: History is repeating itself, a probable zoonotic spillover as a cause of an epidemic: the case of 2019 novel Coronavirus.

text: Pathogen transmission from a vertebrate animal to a human, also known as zoonotic spillover, represents a global public health burden, which while associated with multiple outbreaks, still remains a poorly understood phenomenon. Coronaviruses, like influenza viruses, circulate in nature in various animal species. Alpha-coronaviruses and beta-coronaviruses can infect mammals and gamma-coronaviruses and delta-coronaviruses tend to infect birds, but some of them can also be transmitted to mammals. Although still preliminary, current data suggest that bats are the most probable initial source of the current 2019 novel CoV (2019nCoV) outbreak, that begun on December 2019 in Wuhan, China, apparently spreading from a "wet market" to multiple cities and provinces in China. This epidemic of 2019nCoV, already reaching more than 6,000 cases to-day (end of January 2020) (>90% in China), will not be the last one linked to zoonotic spillover events.


title: [Identification and characterization of new and unknown coronaviruses using RT-PCR and degenerate primers].

text: A modified method of the reverse transcription followed by polymerase chain reaction (RT-PCR) was developed in order to examine the genome of a recently discovered virus, the porcine epidemic diarrhoea virus (PEDV), which resembled morphologically the coronaviruses. The published sequences of the genomes of various coronaviruses were compared. On the level of the amino acid sequence, conserved regions, common to all coronaviruses, were found in the gene encoding the nonstructural protein 1b as well as in the genes coding for the major structural proteins (S, M, and N). Due to the degeneration of the genetic code, some amino acids may be encoded by different nucleotide triplets. In order to compensate for this degeneration, mixtures of primers were synthesized, containing a variety of nucleotide sequences which together represented all possible codons for the conserved amino acid sequences. This method allowed to amplify and clone approximately 4000 base pairs of the genome of PEDV. An analysis of the genomic sequences revealed that PEDV holds an interesting intermediate position between human coronavirus 229E and Transmissible Gastroenteritis virus. We postulate that the method presented in this contribution may be useful to study and characterize other unknown viruses, especially viruses for which no cell cultures for propagation are available.


title: Social Media's Initial Reaction to Information and Misinformation on Ebola, August 2014: Facts and Rumors.

text: OBJECTIVE We analyzed misinformation about Ebola circulating on Twitter and Sina Weibo, the leading Chinese microblog platform, at the outset of the global response to the 2014-2015 Ebola epidemic to help public health agencies develop their social media communication strategies. METHODS We retrieved Twitter and Sina Weibo data created within 24 hours of the World Health Organization announcement of a Public Health Emergency of International Concern (Batch 1 from August 8, 2014, 06:50:00 Greenwich Mean Time [GMT] to August 9, 2014, 06:49:59 GMT) and seven days later (Batch 2 from August 15, 2014, 06:50:00 GMT to August 16, 2014, 06:49:59 GMT). We obtained and analyzed a 1% random sample of tweets containing the keyword Ebola. We retrieved all Sina Weibo posts with Chinese keywords for Ebola for analysis. We analyzed changes in frequencies of keywords, hashtags, and Web links using relative risk (RR) and c(2) feature selection algorithm. We identified misinformation by manual coding and categorizing randomly selected sub-datasets. RESULTS We identified two speculative treatments (i.e., bathing in or drinking saltwater and ingestion of Nano Silver, an experimental drug) in our analysis of changes in frequencies of keywords and hashtags. Saltwater was speculated to be protective against Ebola in Batch 1 tweets but their mentions decreased in Batch 2 (RR=0.11 for "salt" and RR=0.14 for "water"). Nano Silver mentions were higher in Batch 2 than in Batch 1 (RR=10.5). In our manually coded samples, Ebola-related misinformation constituted about 2% of Twitter and Sina Weibo content. A range of 36%-58% of the posts were news about the Ebola outbreak and 19%-24% of the posts were health information and responses to misinformation in both batches. In Batch 2, 43% of Chinese microblogs focused on the Chinese government sending medical assistance to Guinea. CONCLUSION Misinformation about Ebola was circulated at a very low level globally in social media in either batch. Qualitative and quantitative analyses of social media posts can provide relevant information to public health agencies during emergency responses.


title: Isolation and characterization of sialodacryoadenitis virus (coronavirus) from rats by established cell line LBC.

text: The outbreak of sialoadenitis occurred in a laboratory rat colony and the causative agent was isolated from the affected salivary glands of diseased rats using the established cell line LBC. The isolate readily multiplied, producing clear cytopathic effects with syncytium formation, and it was identified virologically and serologically as rat sialodacryoadenitis virus. In attempts to isolate the virus by primary rat kidney (PRK) cells and suckling mice as well as LBC cells, the LBC cells showed higher susceptibility for the virus growth as compared with PRK cells or the brain of suckling mice. The isolation rate of virus was 100% (5/5) in LBC, 40% (2/5) in PRK cells and 60% (3/5) in suckling mice. After four passages in the LBC cells, the virus did not produce disease in adult rats, while the mouse brain-passaged virus did.


On the role of different lymphocyte subpopulations in the course of coronavirus MHV IV (JHM)-induced encephalitis in Lewis rats.


title: Outbreak of human metapneumovirus detected by use of the Vero E6 cell line in isolates collected in Yamagata, Japan, in 2004 and 2005.

text: A number of epidemiological studies have shown human metapneumovirus (hMPV) to be one of the most important viral agents associated with acute respiratory infections in humans. However, due to the difficulty in growing the virus, all epidemiological studies of hMPV infection have been performed on the basis of the molecular method. Thus, the development of a cell line suitable for the isolation of hMPV from clinical specimens is a crucial step for further research. Using the Vero E6 cell line, which could be stably maintained for 1 month without passage or medium change, we succeeded in isolating 79 strains from 4,112 specimens obtained in Yamagata, Japan, in 2004 and 2005. The total isolation rate was 1.9% (79/4,112). The monthly distribution revealed that hMPV infections occurred between February and April in 2004 and throughout most of the year in 2005. Phylogenetic analysis indicated that subgenogroup B2 was predominant in 2004, whereas three subgenogroups, A2, B1, and B2, had cocirculated in 2005. Although multiple subgenogroups cocirculated in 2005, each individual subgenogroup strain was found to predominate at specific sites. An infectivity assay of hMPV strains also indicated that the infection efficiency in Vero E6 cells was better than that in LLC-MK2 cells. Finally, we found that Vero E6 cells are useful for the isolation of hMPVs and that this utility might aid further research into hMPVs beyond the epidemiological data shown in this study.


title: Effect of Right Insular Involvement on Death and Functional Outcome After Acute Ischemic Stroke in the IST-3 Trial (Third International Stroke Trial).

text: BACKGROUND AND PURPOSE In patients with acute ischemic stroke, whether involvement of the insular cortex influences outcome is controversial. Much of the apparent adverse outcome may relate to such strokes usually being severe. We examined the influence of right and left insular involvement on stroke outcomes among patients from the IST-3 study (Third International Stroke Trial) who had visible ischemic stroke on neuroimaging. METHODS We used multiple logistic regression to compare outcomes of left versus right insular and noninsular strokes across strata of stroke severity, on death, proportion dead or dependent, and level of disability (ordinalized Oxford Handicap Score) at 6 months, with adjustment for the effects of age, lesion size, and presence of atrial fibrillation. RESULTS Of 3035 patients recruited, 2099 had visible ischemic strokes limited to a single hemisphere on computed tomography/magnetic resonance scans. Of these, 566 and 714 had infarction of right and left insula. Six months after randomization, right insular involvement was associated with increased odds of death when compared with noninsular strokes on the left side (adjusted odds ratio, 1.83; 95% confidence interval, 1.33-2.52), whereas the adjusted odds ratio comparing mortality after insular versus noninsular strokes on the left side was not significant. Among mild/moderate strokes, outcomes for right insular involvement were worse than for left insular, but among more severe strokes, the difference in outcomes was less substantial. CONCLUSIONS We found an association between right insular involvement and higher odds of death and worse functional outcome. The difference between right- and left-sided insular lesions on outcomes seemed to be most evident for mild/moderate strokes. CLINICAL TRIAL REGISTRATION URL: http://www.isrctn.com. Unique identifier: ISRCTN25765518.