title: Comparison of conventional viral cultures with direct fluorescent antibody stains for diagnosis of community-acquired respiratory virus infections in hospitalized children.

text: OBJECTIVE Because of the widespread availability of rapid viral antigen testing, many institutions never adopted a routine practice of ordering viral cultures to detect community-acquired respiratory viruses (CRVs). The ease of performing complete viral studies in our on site laboratory allowed us to assess the clinical implications of the absence of conventional culture results in previously healthy hospitalized children with CRV infections. METHODS From June 1997 through May 2000, the results of direct immunofluorescence assay (DFA) of 1069 nasopharyngeal swab (NP) specimens were compared with simultaneously inoculated conventional tube cell cultures for detection of CRVs. In addition the medical records of 140 previously healthy infants and children hospitalized for management of lower respiratory tract infections caused by culture-proved CRVs were reviewed. RESULTS Viruses were isolated or detected by DFA or viral culture or both in 468 (30%) of the 1557 NP samples evaluated. The most common CRV isolated was respiratory syncytial virus (49%), followed by parainfluenza viruses (15%), influenza A viruses (14%), rhinoviruses (8%), adenoviruses (4%), enteroviruses (4%) and influenza B viruses (1%). Of the 1069 NP specimens for which both viral culture and rapid antigen testing were performed, 190 specimens were DFA-positive and culture-positive, 7 specimens were DFA-positive and culture-negative, 35 specimens were DFA-negative and culture-positive and 837 specimens were DFA-negative and culture-negative. The overall sensitivity, specificity, positive predictive value and negative predictive value of DFA were 84, 99, 96 and 96%, respectively. Of the 140 hospitalized patients with culture-proved viral cultures (89 respiratory syncytial virus, 22 influenza A, 20 parainfluenza virus and 9 adenovirus), the mean duration of hospital stay was 3.6 days, and the mean time for viral cultures to become positive was 7.7 days (P < 0.001, signed rank test). One hundred twenty (86%) viral cultures did not become positive until after the patient had been discharged from the hospital. In no case was the clinical decision regarding the patient's treatment or discharge from the hospital based on the results of viral culture. CONCLUSIONS We conclude that positive viral cultures have no impact on clinical decision making and management of healthy children during hospitalization for illness attributable to community-acquired respiratory viruses.


Fc receptor-like activity of mouse hepatitis virus E2 glycoprotein.


title: Clinical and laboratory features in the early stage of severe acute respiratory syndrome.

text: BACKGROUND AND PURPOSE To characterize the clinical and laboratory features of severe acute respiratory syndrome (SARS) in the early stage and to compare them with those of patients initially suspected of having SARS who were later determined to have other febrile diseases. METHODS Between March and June 2003, 122 patients with possible SARS were admitted to the isolation ward of Tri-Service General Hospital. SARS was diagnosed according to the modified World Health Organization case definition (May 1, 2003). Among them, 43 were classified as probable SARS cases and a SARS etiology was excluded in 32 patients. RESULTS Presenting symptoms on admission included fever (97.7% of probable cases, 84.4% of excluded cases), chills (39.5% vs 18.8%), cough with sputum production (16.3% vs 40.6%), dry cough (23.3% vs 9.4%), dyspnea (18.6% vs 9.4%), diarrhea (14.0% vs none), rhinorrhea (2.3% vs none), and myalgia (7.0% vs 6.6%). Common laboratory features included lymphopenia and elevated aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, C-reactive protein and creatine kinase values. Intubation and mechanical ventilation were required in 12 probable cases and 6 excluded cases. Five patients with probable SARS (11.6%) died. A scoring system which was developed to differentiate SARS patients from other febrile patients in the emergency room could differentiate probable cases from excluded cases with a sensitivity of 36.4% and a specificity of 70.6%. CONCLUSIONS The clinical presentation and laboratory features at the early stage do not allow differentiation of patients with SARS-CoV infection from other febrile patients. Thus, it is mandatory for all healthcare workers to strictly follow standard isolation precautions during an outbreak to minimize disease transmission.


Promising of spatial-temporal model in public health.


title: Population-based simulations of influenza pandemics: validity and significance for public health policy.

text: OBJECTIVE To examine the validity and usefulness of pandemic simulations aimed at informing practical decision-making in public health. METHODS We recruited a multidisciplinary group of nine experts to assess a case-study simulation of influenza transmission in a Swedish county. We used a non-statistical nominal group technique to generate evaluations of the plausibility, formal validity (verification) and predictive validity of the simulation. A health-effect assessment structure was used as a framework for data collection. FINDINGS The unpredictability of social order during disasters was not adequately addressed by simulation methods; even minor disruptions of the social order may invalidate key infrastructural assumptions underpinning current pandemic simulation models. Further, a direct relationship between model flexibility and computation time was noted. Consequently, simulation methods cannot, in practice, support integrated modifications of microbiological, epidemiological and spatial submodels or handle multiple parallel scenarios. CONCLUSION The combination of incomplete surveillance data and simulation methods that neglect social dynamics limits the ability of national public health agencies to provide policy-makers and the general public with the critical and timely information needed during a pandemic.


title: Murine coronavirus spike protein determines the ability of the virus to replicate in the liver and cause hepatitis.

text: Recombinant mouse hepatitis viruses (MHV) differing only in the spike gene, containing A59, MHV-4, and MHV-2 spike genes in the background of the A59 genome, were compared for their ability to replicate in the liver and induce hepatitis in weanling C57BL/6 mice infected with 500 PFU of each virus by intrahepatic injection. Penn98-1, expressing the MHV-2 spike gene, replicated to high titer in the liver, similar to MHV-2, and induced severe hepatitis with extensive hepatocellular necrosis. S(A59)R13, expressing the A59 spike gene, replicated to a somewhat lower titer and induced moderate to severe hepatitis with zonal necrosis, similar to MHV-A59. S4R21, expressing the MHV-4 spike gene, replicated to a minimal extent and induced few if any pathological changes, similar to MHV-4. Thus, the extent of replication and the degree of hepatitis in the liver induced by these recombinant viruses were determined largely by the spike protein.


docid: uwmkzhwb

title: Matrix protein mutations contribute to inefficient induction of apoptosis leading to persistent infection of human neural cells by vesicular stomatitis virus.

text: In a model system to study factors involved in the establishment of a persistent viral infection that may lead to neurodegenerative diseases, Indiana and New Jersey variants of vesicular stomatitis virus (VSV) with different capacities to infect and persist in human neural cells were studied. Indiana matrix (M) protein mutants and the wild-type New Jersey strain persisted in the human neural cell line H4 for at least 120 days. The Indiana wild-type virus (HR) and a non-M mutant (TP6), both unable to persist, induced apoptosis more strongly than all the other variants tested, as indicated by higher levels of DNA fragmentation and caspase-3-like activity. Transfection of H4 cells with mRNA coding for the VSV M protein confirmed the importance of this protein in the induction of apoptosis. Furthermore, the pan-caspase inhibitor ZVAD-fmk maintained cell survival to about 80%, whereas inhibition of caspase-8, caspase-9, or both only partially protected the cells against death, consistent with the fact that anti-apoptotic molecules from the Bcl-2 family also protect cells from death only partially. These results suggest that VSV activates many pathways of cell death and that an inefficient induction of caspase-3-related apoptosis participates in the establishment of a persistent infection of human neural cells by less virulent VSV variants.


title: Inactivated SARS-CoV vaccine prepared from whole virus induces a high level of neutralizing antibodies in BALB/c mice.

text: We tested the ability of inactivated SARS-CoV vaccine to induce neutralizing antibodies in BALB/c mice. The inactivated vaccine was prepared by SARS-CoV virus propagation in Vero cells, with subsequent beta-propiolactone inactivation and Sepharose 4FF column chromatography purification. One hundred forty BALB/c female mice were divided into seven groups of 20 mice each. Of the seven groups, three groups were inoculated with 0.1, 1, and 3 microg of the vaccine without adjuvant while three other groups were inoculated at the same three dosages of vaccine with aluminum hydroxide as adjuvant, respectively. The remaining group was set up as a blank control. Each mouse was inoculated twice at an interval of 3 weeks. One week after the second immunization, mice sera were collected to detect serum neutralizing antibodies. An assay for determining neutralizing antibody titers was developed. The results can be summarized as follows: (1) higher dosages of vaccine induced higher levels of neutralizing antibody titer; (2) the level of neutralizing antibodies induced by the inoculation with aluminum hydroxide adjuvant was slightly higher than that without adjuvant, but the difference was not statistically significant.


title: [Comparison of severe acute respiratory syndrome with community-acquired pneumonia].

text: OBJECTIVE To study the features of severe acute respiratory syndrome (SARS) and community acquired pneumonia (CAP). METHODS The clinical presentation, chest radiographs, white blood cell count and outcome of 45 SARS patients from April 5 to 20 2003 were compared, with those of 80 CAP patients from October 1 2002 to April 1 2003 in our hospital, and the clinical features of SARS were summarized. RESULTS (1) Fever above 38.5 centigrade, dry cough, short of breath, headache, myalgia, diarrhea in SARS were more common in SARS patients than those in CAP (P<0.01). (2) Leucopenia was more common and leukocytosis was less common in SARS than those in CAP (P<0.01). (3) 22.2 percent of SARS patients were found abnormal chest radiographs within 24 hours after onset, 71.1 percent of SARS patients were showed progressive pulmonary infiltration via serial chest radiographs, while all CAP patients had abnormal chest radiographs when they were seen first time, and no patient showed progressive pulmonary infiltration, the difference was significant between the two groups (P<0.01). (4) Patients with SARS were lack of a response to the initial antimicrobial therapy, all patients with CAP had an adequate clinical response within three days, the difference was significant between the two groups(P<0.01 ). (5) The ratio of severe pneumonia and mortality in SARS was much higher than in CAP (both P<0.01). CONCLUSION The clinical presentation, chest radiographs, white blood cell count, response to initial antimicrobial therapy, and outcome in SARS is much different from CAP, differentiate them will be helpful to early recognition, prompt isolation, and prevention of its spread.


title: Properties of Coronavirus and SARS-CoV-2.

text: were identified beginning with the discovery of SARS-CoV in 2002. With the recent detection of SARS-CoV-2, there are now seven human coronaviruses. Those that cause mild diseases are the 229E, OC43, NL63 and HKU1, and the pathogenic species are SARS-CoV, MERS-CoV and SARS-CoV-2 Coronaviruses (order Nidovirales, family Coronaviridae, and subfamily Orthocoronavirinae) are spherical (125nm diameter), and enveloped with club-shaped spikes on the surface giving the appearance of a solar corona. Within the helically symmetrical nucleocapsid is the large positive sense, single stranded RNA. Of the four coronavirus genera (α,β,γ,δ), human coronaviruses (HCoVs) are classified under α-CoV (HCoV-229E and NL63) and β-CoV (MERS-CoV, SARS-CoV, HCoVOC43 and HCoV-HKU1). SARS-CoV-2 is a β-CoV and shows fairly close relatedness with two bat-derived CoV-like coronaviruses, bat-SL-CoVZC45 and bat-SL-CoVZXC21. Even so, its genome is similar to that of the typical CoVs. SARS-CoV and MERS-CoV originated in bats, and it appears to be so for SARS-CoV-2 as well. The possibility of an intermediate host facilitating the emergence of the virus in humans has already been shown with civet cats acting as intermediate hosts for SARS-CoVs, and dromedary camels for MERS-CoV. Human-to-human transmission is primarily achieved through close contact of respiratory droplets, direct contact with the infected individuals, or by contact with contaminated objects and surfaces. The coronaviral genome contains four major structural proteins: the spike (S), membrane (M), envelope (E) and the nucleocapsid (N) protein, all of which are encoded within the 3' end of the genome. The S protein mediates attachment of the virus to the host cell surface receptors resulting in fusion and subsequent viral entry. The M protein is the most abundant protein and defines the shape of the viral envelope. The E protein is the smallest of the major structural proteins and participates in viral assembly and budding. The N protein is the only one that binds to the RNA genome and is also involved in viral assembly and budding. Replication of coronaviruses begin with attachment and entry. Attachment of the virus to the host cell is initiated by interactions between the S protein and its specific receptor. Following receptor binding, the virus enters host cell cytosol via cleavage of S protein by a protease enzyme, followed by fusion of the viral and cellular membranes. The next step is the translation of the replicase gene from the virion genomic RNA and then translation and assembly of the viral replicase complexes. Following replication and subgenomic RNA synthesis, encapsidation occurs resulting in the formation of the mature virus. Following assembly, virions are transported to the cell surface in vesicles and released by exocytosis.


title: Gill-associated virus of Penaeus monodon prawns: an invertebrate virus with ORF1a and ORF1b genes related to arteri- and coronaviruses.

text: A 20089 nucleotide (nt) sequence was determined for the 5' end of the (+)-ssRNA genome of gill-associated virus (GAV), a yellow head-like virus infecting Penaeus monodon prawns. Clones were generated from a approximately 22 kb dsRNA purified from lymphoid organ total RNA of GAV-infected prawns. The region contains a single gene comprising two long overlapping open reading frames, ORF1a and ORF1b, of 4060 and 2646 amino acids, respectively. The ORFs are structurally related to the ORF1a and ORF1ab polyproteins of coronaviruses and arteriviruses. The 99 nt overlap between ORF1a and ORF1b contains a putative AAAUUUU 'slippery' sequence associated with -1 ribosomal frameshifting. A 131 nt stem-loop with the potential to form a complex pseudoknot resides 3 nt downstream of this sequence. Although different to the G/UUUAAAC frameshift sites and 'H-type' pseudoknots of nidoviruses, in vitro transcription/translation analysis demonstrated that the GAV element also facilitates read-through of the ORF1a/1b junction. As in coronaviruses, GAV ORF1a encodes a 3C-like cysteine protease domain located between two hydrophobic regions. However, its sequence suggests some structural relationship to the chymotrypsin-like serine proteases of arteriviruses. ORF1b encodes homologues of the 'SDD' polymerase, which among (+)-RNA viruses is unique to nidoviruses, as well as metal-ion-binding and helicase domains. The presence of a dsRNA replicative intermediate and ORF1a and ORF1ab polyproteins translated by a-1 frameshift suggests that GAV represents the first invertebrate member of the Order NIDOVIRALES:


title: Public health monitoring tools for multiple data streams.

text: INTRODUCTION In concert with increased concerns regarding both biologic terrorism and new natural infectious disease threats (e.g., severe acute respiratory syndrome [SARS] and West Nile virus), as a result of advances in medical informatics, various data sources are available to epidemiologists for routine, prospective monitoring of public health. The synthesis of this evidence requires tools to find anomalies within various data stream combinations while maintaining manageable false alarm rates. OBJECTIVES The objectives of this report are to establish statistical hypotheses to define the compound multivariate problem of surveillance systems, present statistical methods for testing these hypotheses, and examine results of applying these methods to simulated and actual data. METHODS Canonical problems of parallel monitoring and consensus monitoring are considered in this report. Modified Bonferroni methods are examined for parallel monitoring. Both multiple univariate and multivariate methods are applied for consensus monitoring. A multivariate adaptation of Monte Carlo trials, using the injection of epidemic-curve-like signals in the multiple data streams of interest, is presented for evaluation of the various tests. RESULTS The Monte Carlo test results demonstrate that the multiple univariate combination methods of Fisher and Edgington provide the most robust detection performance across the scenarios tested. As the number of data streams increases, methods based on Hotelling's T2 offer added sensitivity for certain signal scenarios. This potential advantage is clearer when strong correlation exists among the data streams. CONCLUSION Parallel and consensus monitoring tools must be blended to enable a surveillance system with distributed sensitivity and controlled alert rates. Whether a multiple univariate or multivariate approach should be used for consensus monitoring depends on the number and distribution of useful data sources and also on their covariance structure and stationarity. Strong, consistent correlation among numerous sources warrants the examination of multivariate control charts.


title: Virus-induced airway hyperresponsiveness in man.

text: Airway hyperresponsiveness is the most prominent functional abnormality in asthma. Although its aetiology is still unclear, it is well-known that allergen exposure and virus infections can temporarily induce or aggravate airway hyperresponsiveness. Among these environmental factors, virus infections appear to be clinically most relevant, since recent epidemiological studies have shown that most asthma exacerbations in children are associated with positive nasopharyngeal viral identification. The pathogenesis of virus-induced airway hyperresponsiveness has been investigated by experimental virus infections in animals and in man. Intranasal inoculation and/or inhalation of live attenuated influenza virus, or certain strains of rhinovirus, have been shown to induce airway hyperresponsiveness to various bronchoconstrictor stimuli in man. This indicates that experimental virus-infection, like allergen challenge, is an appropriate investigational model of asthma. The mechanisms of virus-induced airway hyperresponsiveness are still unclear, but may, in part, be similar to those involved in the pathogenesis of asthma. Currently investigated hypothesis include: epithelial damage or dysfunction, immunological responses, inflammatory mediator release, cholinergic and/or noncholinergic reflexes, and impaired beta-adrenoceptor function. Careful experimental studies, using modern laboratory techniques, are needed to unravel the role of viruses in the development of airway hyperresponsiveness. The results of such studies can potentially lead to an improvement of future asthma management.


title: Characterization of immune cells and perforin mutations in familiar venous thromboembolism.

text: AIM This study was to carry out exome sequencing in a Han Chinese family with venous thromboembolism. METHODS Three venous thromboembolism (VTE) patients and five members from a Han Chinese family were evaluated by exome sequencing. RESULTS Among the 3 VTE patients, mutations of 2 genes including PRF1 and HTR2A were identified and predicted to be functionally damaged to their encoded proteins. In addition, the PRF1 mutation and the HTR2A mutation identified in our study were absent in 100 non-related controls, indicating that venous thromboembolism has a genetic component. The R357W mutation is located in the membrane attack complex/perforin domain of PRF1 protein, which exists in both the perforin. The steps of killing foreign or pathological antigen cells by NK cells, CD8 (+)T cells and the membrane attack complex include membrane perforation and release of the granzyme, either of which is abnormal can lead to immune dysfunction. CONCLUSIONS The mutations of immune related genes in familial VTE might provide new understanding of the pathogenesis of familial venous thromboembolism.


title: Case-control and two-gate designs in diagnostic accuracy studies.

text: BACKGROUND In some diagnostic accuracy studies, the test results of a series of patients with an established diagnosis are compared with those of a control group. Such case-control designs are intuitively appealing, but they have also been criticized for leading to inflated estimates of accuracy. METHODS We discuss similarities and differences between diagnostic and etiologic case-control studies, as well as the mechanisms that can lead to variation in estimates of diagnostic accuracy in studies with separate sampling schemes ("gates") for diseased (cases) and nondiseased individuals (controls). RESULTS Diagnostic accuracy studies are cross-sectional and descriptive in nature. Etiologic case-control studies aim to quantify the effect of potential causal exposures on disease occurrence, which inherently involves a time window between exposure and disease occurrence. Researchers and readers should be aware of spectrum effects in diagnostic case-control studies as a result of the restricted sampling of cases and/or controls, which can lead to changes in estimates of diagnostic accuracy. These spectrum effects may be advantageous in the early investigation of a new diagnostic test, but for an overall evaluation of the clinical performance of a test, case-control studies should closely mimic cross-sectional diagnostic studies. CONCLUSIONS As the accuracy of a test is likely to vary across subgroups of patients, researchers and clinicians might carefully consider the potential for spectrum effects in all designs and analyses, particularly in diagnostic accuracy studies with differential sampling schemes for diseased (cases) and nondiseased individuals (controls).


title: Y-crossing of braided stents with stents and flow diverters does not cause significant stenosis in bench-top studies.

text: Y-stent placement to treat bifurcation aneurysms requires the second device to cross the confines of the first stent, with concerns regarding the formation of stenosis of the second device at the site of crossing. Various braided stents and flow diverters (FDs) were deployed to cross through a high porosity braided stent, in a Y configuration, with the ends of the devices inserted in plastic tubes of various diameters, leaving the mid-portion free to expand. The ensuing constructs were photographed, paying attention to the degree of stenosis, if any, created where the second device crosses the first stent. Experiments were repeated selecting different zones of the first stent as the site of crossing for the second device, different tube diameters, and changing the angle of the bifurcation. Crossing the first stent did not cause the second stent to become significantly stenosed in any case. Crossing through the transition or expansion zone of the first device had no influence on results. Different bifurcation angles had no influence on the occurrence of stenosis. Y-stent placement to treat arterial bifurcations using braided self-expanding stents and FDs does not lead to significant stenosis in bench-top studies.


title: The responsibility of healthcare institutions to protect global health security.

text: New public threats that can rapidly cross borders are continuing to challenge global health securityand will require unprecedented levels of co-operation. At the international level, the response to this challenge led to the approval of revised International Health Regulations (IHR). This unanimously approved document outlines how countries are to prepare for and respond to public health emergencies of international concern in a manner that does not unnecessarily impact on travel and trade.


title: Characterization of the heptad repeat regions, HR1 and HR2, and design of a fusion core structure model of the spike protein from severe acute respiratory syndrome (SARS) coronavirus.

text: Severe acute respiratory syndrome coronavirus (SARS-CoV) is a newly emergent virus responsible for a worldwide epidemic in 2003. The coronavirus spike proteins belong to class I fusion proteins, and are characterized by the existence of two heptad repeat (HR) regions, HR1 and HR2. The HR1 region in coronaviruses is predicted to be considerably longer than that in other type I virus fusion proteins. Therefore the exact binding sequence to HR2 from the HR1 is not clear. In this study, we defined the region of HR1 that binds to HR2 by a series of biochemical and biophysical measures. Subsequently the defined HR1 (902-952) and HR2 (1145-1184) chains, which are different from previously defined binding regions, were linked together by a flexible linker to form a single-chain construct, 2-Helix. This protein was expressed in Escherichia coli and forms a typical six-helix coiled coil bundle. Highly conserved HR regions between mouse hepatitis virus (MHV) and SARS-CoV spike proteins suggest a similar three-dimensional structure for the two fusion cores. Here, we constructed a homology model for SARS coronavirus fusion core based on our biochemical analysis and determined the MHV fusion core structure. We also propose an important target site for fusion inhibitor design and several strategies, which have been successfully used in fusion inhibitor design for human immunodeficiency virus (HIV), for the treatment of SARS infection.


title: CXC chemokine ligand 10 controls viral infection in the central nervous system: evidence for a role in innate immune response through recruitment and activation of natural killer cells.

text: How chemokines shape the immune response to viral infection of the central nervous system (CNS) has largely been considered within the context of recruitment and activation of antigen-specific lymphocytes. However, chemokines are expressed early following viral infection, suggesting an important role in coordinating innate immune responses. Herein, we evaluated the contributions of CXC chemokine ligand 10 (CXCL10) in promoting innate defense mechanisms following coronavirus infection of the CNS. Intracerebral infection of RAG1(-/-) mice with a recombinant CXCL10-expressing murine coronavirus (mouse hepatitis virus) resulted in protection from disease and increased survival that correlated with a significant increase in recruitment and activation of natural killer (NK) cells within the CNS. Accumulation of NK cells resulted in a reduction in viral titers that was dependent on gamma interferon secretion. These results indicate that CXCL10 expression plays a pivotal role in defense following coronavirus infection of the CNS by enhancing innate immune responses.


title: Genetic evidence for a structural interaction between the carboxy termini of the membrane and nucleocapsid proteins of mouse hepatitis virus.

text: The coronavirus membrane (M) protein is the most abundant virion protein and the key component in viral assembly and morphogenesis. The M protein of mouse hepatitis virus (MHV) is an integral membrane protein with a short ectodomain, three transmembrane segments, and a large carboxy-terminal endodomain facing the interior of the viral envelope. The carboxy terminus of MHV M has previously been shown to be extremely sensitive to mutation, both in a virus-like particle expression system and in the intact virion. We have constructed a mutant, M(Delta)2, containing a two-amino-acid truncation of the M protein that was previously thought to be lethal. This mutant was isolated by means of targeted RNA recombination with a powerful host range-based selection allowed by the interspecies chimeric virus fMHV (MHV containing the ectodomain of the feline infectious peritonitis virus S protein). Analysis of multiple second-site revertants of the M(Delta)2 mutant has revealed changes in regions of both the M protein and the nucleocapsid (N) protein that can compensate for the loss of the last two residues of the M protein. Our data thus provide the first genetic evidence for a structural interaction between the carboxy termini of the M and N proteins of MHV. In addition, this work demonstrates the efficacy of targeted recombination with fMHV for the systematic genetic analysis of coronavirus structural protein interactions.


title: Passive immunity to transmissible gastroenteritis virus: intramammary viral inoculation of sows.

text: Sows were injected intramammarily with live-attenuated TGE virus, an enteric coronavirus--one sow during pregnancy and three sows during lactation. All sows were TGE antibody seronegative prior to inoculation except for one naturally infected sow inoculated during lactation. The animal injected during pregnancy had primarily IgG TGE antibodies in milk from all glands. By contrast, sows injected during lactation had IgA and IgM initially, and later IgA and IgG TGE antibodies in milk from injected and noninjected glands. The seropositive sow had elevated IgA TGE antibody titers in milk after IMm injection. Both seronegative sows inoculated intramammarily during lactation shed TGE virus in milk from injected glands, and their nursing piglets developed mild diarrhea and shed virus in their feces at three to nine DPE of the sows. Milk from IMm injected glands generally had higher TGE antibody titers than milk from noninjected glands. These results suggest that TGE virus replicates in lactating mammary gland tissue, thereby stimulating IgA immunocytes, leading to secretion of IgA antibodies in milk. Whether the intramammary route presents a natural route of enteric virus exposure in lactating animals (by way of infected nursing piglets), leading to IgA-antibody secretion in milk, requires further investigation.


title: Alveolar epithelial lining fluid cellularity, protein and endothelin-1 in children with congenital heart disease.

text: This study applied bronchoalveolar lavage (BAL) to children with congenital heart disease (CHD) prior to elective cardiac catheterization (n = 48), to determine the influence of pulmonary blood flow and viral infection on the alveolar epithelial lining fluid (ELF) concentration of leucocytes, protein and endothelin-1 (ET-1). Lower respiratory tract (LRT) viral infection was defined as either a positive immunofluorescence for virus, or a virus cultured from the bronchoalveolar lavage fluid (BALF). Haemodynamic status was determined at cardiac catheterization. Normative data for BALF, but not ELF parameters, were obtained from 26 asymptomatic, noninfected normal children undergoing elective surgery. In the absence of LRT infection, the BALF macrophage, lymphocyte and neutrophil differential in CHD was not significantly different from the normal controls. In CHD, both increased pulmonary-to-systemic flow ratio (Q'p/Q's) and increased pulmonary artery-to-left ventricular pressure ratio PAP/LVP were associated with a decrease in ELF protein (rs = -0.59; p < 0.0001; and rs = -0.50; p < 0.0001 respectively). A respiratory virus was isolated from the BALF in 8 (17%) of CHD children. Virus isolation was associated with an increased ELF total protein (p < 0.05 vs no infection), a decreased alveolar macrophage differential count (p < 0.01), and an increased neutrophil differential count (p < 0.05). ET-1 was detected in the BALF of 83% of the noninfected CHD children compared to only 23% of the controls (p < 0.001). ELF ET-1 concentrations did not correlate with haemodynamic status in CHD, but were up to 100 times higher than paired plasma levels. We conclude that, in congenital heart disease, both lower respiratory tract viral infection and increased pulmonary blood flow and/or pulmonary vascular pressure influence the alveolar milieu. High alveolar epithelial lining fluid concentrations of endothelin-1 occur in congenital heart disease, but the stimulus for pulmonary endothelin-1 production is unclear.


docid: fmfqybyz

title: [Forhøjet D-dimer og karakteristiske billeddiagnostiske fund hos patient med COVID-19].


title: Nondisruptive detection of activity of catabolic promoters of Pseudomonas putida with an antigenic surface reporter system.

text: A simple procedure to detect the switching on and off of catabolic promoters of Pseudomonas putida, at the level of single cells based on the immunodetection of a reporter epitope expressed on the surface of bacterial cells, has been developed. To do this, the antigenic sequence Asp-Leu-Pro-Pro-Asn-Ser-Asp-Val-Val-Asp, from a coronavirus, was inserted genetically in the permissive site around amino acid position 153 of the LamB protein (maltose and lambda phage receptor) of Escherichia coli. When the hybrid lamB gene is transcribed, the epitope becomes presented on the surface of the bacterial cells in a configuration available to specific antibodies. To validate this notion in nonenteric bacteria, the expression and correct processing of LamB were confirmed by coupling the lamB gene to the salicylate-responsive Psa1 promoter of the NAH7 (naphthalene degradation) plasmid in Pseudomonas putida. Subsequently, a hybrid lamB gene carrying the sequence of the coronavirus antigen was placed downstream of the m-toluate-responsive Pm promoter of the TOL (toluene degradation) plasmid. Exposure of the epitope on the Pseudomonas cell surface was monitored through fluorescence of whole cells treated with a monoclonal antibody against the heterologous antigen. Fluorescence emission was dependent on the presence of m-toluate in the medium, thus permitting detection of the Pm promoter switching on by simple optical inspection of individual cells, even in situations when these are a very minor component of a complex bacterial community.


Are patients with systemic lupus erythematosus more prone to result false-positive for SARS-CoV2 serology?


title: [COVID-19 experience in a private institution in Buenos Aires during the first month of the pandemic: 26 cases].

text: The first case of coronavirus (COVID-19) infection in Argentina was reported in Buenos Aires on March 3rd, 2020, in a patient who arrived from Milan. The aim of this study is to describe the clinical characteristics and course of illness of patients infected with Covid-19 that were hospitalized in a private clinical setting during March 2020. Sixty three patients suspected of COVID-19 infection, were admitted to our hospital during March 2020. Twenty six of these subjects were diagnosed positive by the RT-PCR for SARS Cov-2 in a nasopharyngeal swab specimen. We recorded data about epidemiological characteristics, clinical signs and symptoms, in addition to comorbidities, laboratory and radiological studies were done. The median age was 40 years (SD ± 15 years); there was an equal number of men and women; all the patients but one were coming from abroad; the most common symptom was fever (24/26 cases) and lymphopenia (n = 8). We found that there were patients in whom fever disappeared within the first 72h (n = 17) and another group in which the fever lasted six or more days (n = 7). One patient out of 26 died. The population of our study was young and almost all were imported cases, in alinement with the stage of the epidemic at the time when the study was conducted. We observed that there were two patterns in the fever curve but the persistence of it did not lead to a worse prognosis. Larger population studies are required in order to confirm these clinical findings.


title: Critical role of nosocomial transmission in the toronto sars outbreak.

text: We develop a compartmental mathematical model to address the role of hospitals in severe acute respiratory syndrome ( SARS ) transmission dynamics, which partially explains the heterogeneity of the epidemic. Comparison of the effects of two major policies, strict hospital infection control procedures and community-wide quarantine measures, implemented in Toronto two weeks into the initial outbreak, shows that their combination is the key to short-term containment and that quarantine is the key to long-term containment.


docid: bdvcwaau

title: What does isolation mean in home healthcare?

text: In the event of an infectious disease outbreak, home health nurses are called upon to care for patients who have been placed on isolation precautions. To protect themselves, patients, and caregivers, nurses must be knowledgeable about infectious diseases and infection control procedures. A checklist is provided to assist the nurse and caregiver with guidance in performing the patient's care.


title: Update: influenza activity--United States, 2003-04 season.

text: Influenza began circulating in the United States unusually early this season, and influenza activity nationwide is expected to increase. Cases of severe disease, including deaths, have been reported in children. This report summarizes influenza activity in the United States during the weeks ending October 4-December 6, 2003. During the week ending December 6, influenza activity was reported to CDC as widespread in 24 states. The early season and the unusually high and persistent demand for vaccine have resulted in a decreasing supply of trivalent inactivated vaccine. Emphasis should be placed on vaccinating persons at high risk for complications from influenza, including healthy children aged 6-23 months. Healthy persons aged 5-49 years who wish to receive vaccine should consider being vaccinated with the intranasally administered live, attenuated influenza vaccine (LAIV), a substantial supply of which remains available.


title: [Necrotizing enterocolitis. A study of forty-six cases seen in a maternity hospital].

text: Forty-six cases of necrotizing enterocolitis were seen at the Baudelocque maternity hospital from august 1978 to october 1980. In eighteen cases, clinical signs associated with roentgenograms showing pneumatosis led to unequivocal diagnosis. A surgical procedure was done in nine of these patients, during the acute stage in four and later on in five. Diagnosis was strongly suspected in twenty-eight cases. Clinical features were less serious and included abdominal distension, bloody stools, and emesis. There were no roentgenological signs. None of these infants underwent surgery. Overall mortality rate was 7% (3/46). In comparison to previously published studies, our series shows lower perinatal risk factors, higher mean birthweight, and lower prematurity rate (14%). No evidence was found to support the responsibility of a specific bacterial agent. Corona virus was found in the stools of five out of eleven cases studied during the second epidemic wave in october 1979, suggesting a possible viral etiology. Several preventive steps have been taken. Breast-feeding has been encouraged. Each mother-infant pair has been isolated instead of grouping infants in night nurseries. Staff members have been given specific information on the means of preventing contamination.


title: Severe respiratory failure in an immunocompetent host with invasive pulmonary aspergillosis and H1N1 influenza.

text: Invasive pulmonary aspergillosis (IPA) is a life-threatening condition that usually occurs in immunocompromised hosts. However, according to recent reports it can affect immunocompetent hosts with severe influenza infection due to viral-dependent disruption of respiratory immune defenses. We present the case of a 61-year-old Caucasian man admitted to the Emergency Department with respiratory failure and fever, who was diagnosed with H1N1 influenza and IPA. Because of his poor general conditions, he was treated with a double antifungal scheme, although this lies outside the suggested treatment guidelines. This choice turned out to be extremely effective. He was discharged after one month and his clinical conditions showed rapid improvement, with nearly complete normalization of the radiological pattern in three months. IPA remains a life-threatening condition, even in immunocompetent hosts, and should therefore always be suspected; if necessary, a combined treatment should rapidly be started. We report this case as the interest in influenza-associated IPA is high, both due to the clinical severity of this condition, which is treatable if identified early, and the emerging importance of respiratory infections caused by viruses belonging to the SARS family, such as SARS-CoV-2.


title: The nature of international health security.

text: Health issues occasionally intersect security issues. Health security has been viewed as an essential part of human security. Policymakers and health professionals, however, do not share a common definition of health security. This article aims to characterize the notions of health security in order to clarify what constitutes the nexus of health and security. The concept of health security has evolved over time so that it encompasses many entities. Analyzing the health reports of four multilateral organizations (the United Nations, World Health Organization, Asia-Pacific Economic Cooperation, and the European Union) produced eight categories of most significant relevance to contemporary health security, allowing comparison of the definitions. The four categories are: emerging diseases; global infectious disease; deliberate release of chemical and biological materials; violence, conflict, and humanitarian emergencies. Two other categories of common concern are natural disasters and environmental change, as well as chemical and radioactive accidents. The final two categories, food insecurity and poverty, are discussed less frequently. Nevertheless, food security is emerging as an increasingly important issue in public health. Health security is the first line of defence against health emergencies. As globalization brings more complexities, dealing with the increased scale and extent of health security will require greater international effort and political support.


title: Crossing the boundaries.

text: In the past, research on zoonotic diseases has been fragmented. Teresa Belcher and Diane Newell discuss the formation of Med-Vet-Net, a "virtual institute", which aims to integrate activity between researchers in human and veterinary medicine across the European Union.


title: In vitro and biodistribution examinations of Tc-99m-labelled doxorubicin-loaded nanoparticles.

text: BACKGROUND Nanoparticles represent promising drug carrier systems. In the case of cytostatics such as doxorubicin, carrier colloid systems as human serum albumin (HSA) nanoparticles, may increase their therapeutic efficiency and decrease their side-effects (toxicity) and any potential multidrug resistance. In the present study, doxorubicin, as a widely used antineoplastic agent, was incorporated into the matrix of human serum albumin and three different particle-sized doxorubicin-loaded HSA nanoparticles were prepared, using a previously described desolvation method. Our objective was to find out if different particle sizes of colloid carriers can allow regarding the given cytostatic agent. MATERIAL AND METHODS The three prepared nanoparticles were labelled using technetium (Tc-99m) and were tested for their physicochemical colloidal quality, fluctuations, and radiochemical stability. Biodistribution of different-sized radiolabelled colloids were determined by means of scintigraphic imaging studies in healthy male Wistar rats. Images were taken by gamma camera at several times and organ uptakes were estimated by quantitative ROI analysis. RESULTS In vitro measurements showed that more than 95% of doxorubicin proportion was permanently adsorbed to human serum albumin. Radiolabelled doxorubicin-loaded particles had high-degree and durable labelling efficiency and particle size stability. Biodistribution results had a close correlation to earlier described results of radiocolloids in similar particle size ranges. In vivo examinations verified that colloid carriers have insignificant size fluctuations after an intravenous application and they show the proper distribution according to their particle size. CONCLUSIONS Our investigations verified that different and stable particle sizes make drug carrier HSA nanoparticles possible to apply different drug targeting in a potential clinical use.


title: Acute and subacute demyelination induced by mouse hepatitis virus strain A59 in C3H mice.

text: Mouse hepatitis virus (MHV, strain A-59), a coronavirus, induced acute and subacute demyelinating disease in the central nervous system of weanling C3H mice that were reported to be genetically resistant to MHV. Subtle clinical signs developed in greater than 90% of animals 5-7 days after intracerebral inoculation of 25 PFU, and foci of demyelination were detected from 1 to 4 weeks postinoculation (p.i.) by histopathology and immunolabeling with antibody directed against myelin basic protein. Infectious virus could be isolated only during the first 10 days p.i., but viral antigens persisted in the cytoplasm of oligodendrocytes in the demyelinating lesions for up to 4 weeks p.i. Serum antibody titers against MHV-A59 rose by 2 weeks p.i. and persisted at high levels for months. This infection of genetically "resistant" animals by a normally hepatotropic strain of MHV can serve as a highly reproducible model for the study of acute and subacute demyelination by a coronavirus.


title: Experimental biomechanical evaluation of polypropylene prostheses used in pelvic organ prolapse surgery.

text: INTRODUCTION AND HYPOTHESIS Although polypropylene (PP) is the most common biomaterial used to repair genital prolapse via vaginal route, its mechanical properties however remain obscure. METHODS An abdominal hernia rabbit model was used to evaluate retraction, solidity, and elasticity of the principal types of PP prostheses currently available, i.e., three large pore size monofilament prostheses, one heavy weight (HWPP), a second low weight (LWPP), and a third coated with atelocollagen (CPP). A small pore size multifilament PP (MPP) implant was also tested. RESULTS In comparison with HWPP (12%), LWPP (15%), and MPP (30%), CPP had less retraction (8% of the original size). Unlike pore size, weight prosthesis is not an influencing factor for retraction. Atelocollagen coating reduced retraction. HWPP and MPP were the most solid prostheses. MPP supported the greatest elastic force. CONCLUSIONS When the biomechanical parameters were comparatively assessed, HWPP was considered to have the most advantageous properties for prolapse surgery.


title: Insertion of a new transcriptional unit into the genome of mouse hepatitis virus.

text: The subgenomic mRNAs of the coronavirus mouse hepatitis virus (MHV) are composed of a leader sequence, identical to the 5' 70 nucleotides of the genome, joined at distant downstream sites to a stretch of sequence that is identical to the 3' end of the genome. The points of fusion occur at intergenic sequences (IGSs), loci on the genome that contain a tract of sequence homologous to the 3' end of the leader RNA. We have constructed a mutant of MHV-A59 containing an extra IGS inserted into the genome immediately downstream of the 3'-most gene, that encoding the nucleocapsid (N) protein. We show that in cells infected with the mutant, there is synthesis of an additional leader-containing subgenomic RNA of the predicted size. Our study demonstrates that (i) an IGS can be a sufficient cis-acting element to dictate MHV transcription, (ii) the relative efficiency of an IGS must be influenced by factors other than the nucleotides immediately adjacent to the 5'AAUCUAAAC3' core consensus sequence or its position relative to the 3' end of the genome, (iii) a downstream IGS can exert a polar attenuating effect on upstream IGSs, and (iv) unknown factors prevent the insertion of large exogenous elements between the N gene and the 3' untranslated region of MHV. These results confirm and extend conclusions previously derived from the analysis of defective interfering RNAs.


title: [A case-control study on the risk factors of severe acute respiratory syndromes among health care workers].

text: OBJECTIVE To study the factors in relation to severe acute respiratory syndromes (SARS) among health care workers and to develop related protective measures. METHODS Case-control study was applied. A standardized questionnaire was used to collect SARS related information for health care workers who had contacted or treated SARS patients. Univariate analysis was conducted using SPSS 10.0 software package and multivariate logistic regression analysis was conducted using SAS 6.12. RESULTS Twenty-seven of the 49 factors under study were significantly associated with SARS infection, in which 22 factors were protective, and the other 5 were risk factors. 27 factors were included for multivariate logistic regression analysis. Results showed that six factors as wearing eye glasses, wearing protection gowns, exposure to secrets/mode of contact with SARS patients, types of mask and the working years atc, remained significant association with hospital infection of SARS. CONCLUSION SARS infection in heath care workers was related to many factors during the process of diagnoses and/or treatment. It is recommended that adequate masks, eye-protection and protective gowns should be adopted for heath care workers during the process of clinical diagnoses and treatment of SARS patients.


title: Pathophysiologic changes due to coronavirus-induced diarrhea in the calf.

text: Extensive water, sodium, chloride, bicarbonate, and potassium losses occur in the diarrheal calf. The water loss is entirely from the extracellular space. In severe cases, hypovolemic shock occurs with the blood volume decreased by as much as one-half. Acidosis, which results from fecal bicarbonate loss, lactic acidosis, and renal dysfunction, results in tissue buffering, which in turn causes the efflux of cellular potassium ions. Although there is a total body potassium deficit, plasma potassium concentration is increased. This, in conjunction with an intracellular deficit, causes weakness, lethargy, and potassium cardiotoxicosis resulting in death. Hypoglycemia also contributes to the weakness and lethargy seem as calves become moribund. These losses from the body and shifts in fluids and electrolytes must be understood to develop the most effective rationale for supportive therapy.


title: Identifying epitopes responsible for neutralizing antibody and DC-SIGN binding on the spike glycoprotein of the severe acute respiratory syndrome coronavirus.

text: The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) uses dendritic cell-specific ICAM-3 grabbing nonintegrin (DC-SIGN) to facilitate cell entry via cellular receptor-angiotensin-converting enzyme 2. For this project, we used recombinant baculoviruses expressing different lengths of SARS-CoV spike (S) protein in a capture assay to deduce the minimal DC-SIGN binding region. Our results identified the region location between amino acid (aa) residues 324 to 386 of the S protein. We then generated nine monoclonal antibodies (MAbs) against the S protein to map the DC-SIGN-binding domain using capture assays with pseudotyped viruses and observed that MAb SIa5 significantly blocked S protein-DC-SIGN interaction. An enhancement assay using the HKU39849 SARS-CoV strain and human immature dendritic cells confirmed our observation. Data from a pepscan analysis and M13 phage peptide display library system mapped the reactive MAb SIa5 epitope to aa residues 363 to 368 of the S protein. Results from a capture assay testing three pseudotyped viruses with mutated N-linked glycosylation sites of the S protein indicate that only two pseudotyped viruses (N330Q and N357Q, both of which lost glycosylation sites near the SIa5 epitope) had diminished DC-SIGN-binding capacity. We also noted that MAb SIb4 exerted a neutralizing effect against HKU39849; its reactive epitope was mapped to aa residues 435 to 439 of the S protein. We offer the data to facilitate the development of therapeutic agents and preventive vaccines against SARS-CoV infection.


title: Oral vaccination: where we are?

text: As early as 900 years ago, the Bedouins of the Negev desert were reported to kill a rabid dog, roast its liver and feed it to a dog-bitten person for three to five days according to the size and number of bites [1] . In sixteenth century China, physicians routinely prescribed pills made from the fleas collected from sick cows, which purportedly prevented smallpox. One may dismiss the wisdom of the Bedouins or Chinese but the Nobel laureate, Charles Richet, demonstrated in 1900 that feeding raw meat can cure tuberculous dogs - an approach he termed zomotherapy. Despite historical clues indicating the feasibility of oral vaccination, this particular field is notoriously infamous for the abundance of dead-end leads. Today, most commercial vaccines are delivered by injection, which has the principal limitation that recipients do not like needles. In the last few years, there has been a sharp increase in interest in needle-free vaccine delivery; new data emerges almost daily in the literature. So far, there are very few licensed oral vaccines, but many more vaccine candidates are in development. Vaccines delivered orally have the potential to take immunization to a fundamentally new level. In this review, the authors summarize the recent progress in the area of oral vaccines.


title: Ethical issues in pandemic planning.

text: In the event of an influenza pandemic, many ethical issues will arise in terms of health risks, resource allocation, and management decisions. Planning decisions may be controversial, such as rationing of antivirals, resource allocation (including hospital beds and vaccinations), occupational risk, rostering of staff, responsibilities of health care workers, quarantine measures, and governance issues. A clear ethical framework is needed to enable understanding of the decision-making process and optimise acceptance of decisions by health care workers and other members of an affected community. Planning decisions need to start being examined now, and will require input from a broad group of experts: health care providers, infrastructure managers, lawyers, ethicists, public health physicians, and community members. The process will need to be open, honest and dynamic.


docid: qehmiorz

title: Functional characterization of CD8+ lymphocytes during coronavirus MHV IV induced encephalitides in rats.


title: Interferon gamma potentiates human coronavirus OC43 infection of neuronal cells by modulation of HLA class I expression.

text: HCN-1A, a human cerebral cortical neuron cell line, was examined for its susceptibility to human coronaviruses. The 229e strain replicated efficiently, but the OC43 strain did not replicate well, if at all. Treatment of the cells with interferon gamma at 20U/ml for 48 hr markedly increased the susceptibility of the cells to infection with OC43 virus as shown by a 100-fold increase in secretion of infectious virus over a four day period as compared to untreated controls. The increased susceptibility was shown to be due to membrane expression of HLA class I by receptor-blockade with a monoclonal antibody specific for HLA molecules.


WHAT'S your diagnosis?


title: Correlation of Caveolin-1 With Vascular Intimal Thickness for Different Locations of Catheter Tips in a Dog Model.

text: INTRODUCTION Many studies have reported increased intimal thickness around the catheter tip after catheterization. Caveolin-1 is a protein in the endothelial cell that acts as a shear sensor causing vascular remodeling. This study aimed to elucidate the suitability of different catheter locations and determine the role of caveolin-1 in canine models. MATERIALS AND METHODS Tunneled silicone 14.5-F catheters were inserted into the left jugular vein and right femoral vein in 8 dogs. The dogs were separated into 2 groups by catheter location and were followed up for 28 days. All dogs underwent extracorporeal circulation 3 times a week. After animal sacrifice, histological and immunohistochemical assays were performed to measure specific cell populations. RESULTS There were higher catheter dysfunction rates and lower blood flow rates in the right femoral vein group compared to the left jugular vein group. There was intimal hyperplasia around the catheter tip in both groups with no significant difference between the two groups. There were caveolin-1 expression in the intimal layer of venous wall around the catheter tip location sites in both groups. CONCLUSIONS These findings indicate that different catheter tip locations may influence catheter function and specific targeting of caveonlin-1 could be a strategy of possible future novel therapies for catheter-related vein stenosis.


title: [Specific antibodies against the nucleocapsid protein and S1 domain of spike glycoprotein in patients with severe acute respiratory syndrome].

text: OBJECTIVE To explore the laws of the appearance of the specific serum antibodies against the nucleocapsid (N) protein and the S1 domain of spike (S) glycoprotein in patients with severe acute respiratory syndrome (SARS) and to evaluate the value of these two proteins to be used as diagnostic makers for SARS. METHODS The serum samples of 86 patients with SARS confirmed clinically and serologically, 31 males and 55 females, aged 9 approximately 86, with the course of disease of 1 approximately 81 days, and 745 healthy persons were collected during the course. The specific immunoglobulin G (IgG) against N protein, IgG against S1 domain of S protein, and the SARS-CoV IgG in these sera were detected by ELISA. RESULTS The positive rates of anti-N-IgG, anti S1-IgG, and anti-SARS-CoV IgG were 14% (6/44), 5% (2/44), and 14% (6/44) respectively in the first week of the course of disease, 56% (10/18), 39% (7/18), and 56% (10/18) respectively in the second week, and 100% (24/24), S1-IgG 83% (20/24), and 100% (24/24) respectively in the third week. The agreement rates of the results of anti-N-IgG and anti-s1-IgG with that of the anti-SARS-CoV IgG were 88% (76/86) and 83% (71/86) respectively. The anti-SARS-CoV IgG positive rate in the healthy persons was 1.88% (14.745). CONCLUSION The N protein and S1 protein may be used as antigens in the development of serological assay for detection of SARS-CoV infection.


title: [Replication and transmission mechanisms of highly pathogenic human coronaviruses].

text: The three known human highly pathogenic coronaviruses are severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus, (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Human highly pathogenic coronaviruses are composed of non-structural proteins, structural proteins and accessory proteins. Viral particles recognize host receptors via spike glycoprotein (S protein), enter host cells by membrane fusion, replicate in host cells through large replication-transcription complexes, and promote proliferation by interfering with and suppressing the host's immune response. Human highly pathogenic coronaviruses are hosted by humans and vertebrates. Viral particles are transmitted through droplets, contact and aerosols or likely through digestive tract, urine, eyes and other routes. This review discusses the mechanisms of proliferation and transmission of highly pathogenic human coronaviruses based on the results of existing research, providing basis for future study on interrupting the transmission and pathogenicity of human highly pathogenic coronaviruses.


Summaries for patients. Characteristics and outcomes of patients with severe acute respiratory syndrome (SARS) in Hong Kong.


title: Hybrid Transvaginal NOTES and Mini-Laparoscopic Colectomy: Benefit Through Synergy.

text: BACKGROUND AND OBJECTIVES Hybrid-natural orifice surgery combines the advantages of traditional transabdominal laparoscopic surgery, while limiting surgical trauma to the abdominal wall. Among various routes of intra-abdominal access, the transvaginal method is most appealing because of its utility and proven safety. We describe a series of 4 colonic resections performed with this approach, combined with minilaparoscopy and needlescopic approaches, and discuss the technical aspects, efficacy, and applicability of this technique. METHODS Three patients were selected to undergo hybrid transvaginal natural-orifice right hemicolectomy. A fourth patient, who underwent a segmental resection of a splenic flexure carcinoma, was included. Transvaginal port access was obtained via posterior colpotomy, and was used for dissection, vascular ligation, bowel division, and anastomosis. We used a combination of standard laparoscopic, minilaparoscopic, and needlescopic instruments transabdominally, focusing on reduced size and number of access points. RESULTS Duration of laparoscopy, oncologic outcomes and rate of operative morbidity were comparable to the published literature. Early return of gastrointestinal function and low analgesic requirements was observed in all patients. No morbidity related to transvaginal access was observed and the procedure was performed without difficulty in all cases. CONCLUSION Colonic resection performed by hybrid natural-orifice technique offers several advantages over purely transabdominal laparoscopic procedures. Transvaginal access is easy to perform and offers excellent safety, efficacy, and versatility, especially for right hemicolectomy. Techniques to reduce abdominal wall surgical trauma, such as minilaparoscopy and needlescopic graspers, can be combined effectively in colonic resections, and may act synergistically to reduce postoperative pain and improve outcomes.


title: SARS CoV main proteinase: The monomer-dimer equilibrium dissociation constant.

text: The SARS coronavirus main proteinase (SARS CoV main proteinase) is required for the replication of the severe acute respiratory syndrome coronavirus (SARS CoV), the virus that causes SARS. One function of the enzyme is to process viral polyproteins. The active form of the SARS CoV main proteinase is a homodimer. In the literature, estimates of the monomer-dimer equilibrium dissociation constant, KD, have varied more than 65,0000-fold, from <1 nM to more than 200 microM. Because of these discrepancies and because compounds that interfere with activation of the enzyme by dimerization may be potential antiviral agents, we investigated the monomer-dimer equilibrium by three different techniques: small-angle X-ray scattering, chemical cross-linking, and enzyme kinetics. Analysis of small-angle X-ray scattering data from a series of measurements at different SARS CoV main proteinase concentrations yielded KD values of 5.8 +/- 0.8 microM (obtained from the entire scattering curve), 6.5 +/- 2.2 microM (obtained from the radii of gyration), and 6.8 +/- 1.5 microM (obtained from the forward scattering). The KD from chemical cross-linking was 12.7 +/- 1.1 microM, and from enzyme kinetics, it was 5.2 +/- 0.4 microM. While each of these three techniques can present different, potential limitations, they all yielded similar KD values.


title: Immunodeficiency associated with multiple concurrent infections in captive Pallas' cats (Otocolobus manul).

text: Five neonatal Pallas' cats (Otocolobus manul) at the Oklahoma City Zoo died from toxoplasmosis with concurrent herpesvirus infection. These multiple infections suggested underlying immunodeficiency, perhaps caused by concurrent infection with feline immunodeficiency virus (FIV); so blood samples were collected for serology, serum protein electrophoresis, lymphocyte proliferation assays, and cytokine analysis by reverse transcriptase-quantitative competitive polymerase chain reaction (RT-qcPCR). Resulting data were compared with data from FIV-infected and control domestic short-haired cats. In addition, peripheral blood mononuclear cell cultures were propagated to detect FIV virus by both RT-qcPCR and detection of reverse transcriptase activity. Serum protein electrophoresis showed that four of six Pallas' cats had increased alpha, globulins. At least two Pallas' cats had decreased lymphoproliferation responses to mitogen. and all three tested animals exhibited defective interleukin-12 gene expression. Although these clinical and laboratory findings suggested an immunodeficiency syndrome, FIV infection could not be confirmed. On the basis of repeated blood test results, it can be concluded that nutritional, metabolic, or other systemic problems probably did not contribute to the disease syndrome. Further investigation of other possible causes of immunodeficiency, including a possible genetic component, in this population is needed.


title: Monoclonal antibody capture enzyme-linked immunosorbent assay for detection of bovine enteric coronavirus.

text: Monoclonal antibodies reactive with three different viral polypeptides were evaluated singly and in combination as the capture antibody(s) in an enzyme-linked immunosorbent assay system for the detection of bovine enteric coronavirus. Similar levels of sensitivity were found for all combinations tested. A sensitive, highly specific, and reproducible assay for the detection of bovine enteric coronavirus was developed, using a mixture of two of these monoclonal antibodies reactive with antigenic components either external or internal to the virion. These monoclonal antibodies were bound indirectly to 96-well plates via rabbit anti-mouse immunoglobulin. After sample application and incubation, virus was detected by using rabbit anti-coronavirus peroxidase conjugate followed by enzyme substrate and chromagen. Fecal samples from a single herd of cows were screened for the presence of coronavirus by this assay. Five percent of clinically normal cows were found to be shedding coronavirus.


title: The role of epithelial injury and repair in the origins of asthma.

text: PURPOSE OF REVIEW We currently understand little about the mechanisms that lead to asthma. The bronchial epithelium is the first cell layer of contact with the environment and as such is an especially attractive target in which to identify novel mechanisms and new therapeutic strategies in disease development. We discuss the role of epithelial injury and wound repair in the origins of asthma. RECENT FINDINGS The presence of inflammation, thickening of the basement membrane and angiogenesis have been described in bronchial biopsies from asthmatic children. We and others have demonstrated the utility of bronchial brushings from children for the isolation, characterization and culture of primary epithelial cells. The results of these experiments suggest that intrinsic differences exist between asthmatic and nonasthmatic epithelial cells. SUMMARY It is becoming increasingly clear from studies involving adults and, more recently, children, that the epithelium orchestrates inflammatory and remodeling responses of the airway. Equally clear is that the asthmatic epithelium responds inappropriately to challenge and displays signs of dysregulated repair. Understanding the regulatory mechanisms involved in these processes, including the role of resident/recruited progenitor cells, is crucial if we are to halt the progression of asthma when the disease first manifests in childhood.


title: Expression of the RNA genome of an animal virus in Saccharomyces cerevisiae.

text: The nucleocapsid of vesicular stomatitis virus (VSV) was introduced into the cytoplasm of Saccharomyces cerevisiae by low pH-dependent fusion of the viral envelope with the spheroplast plasma membrane. This led to de novo synthesis of the three major structural proteins of the virus--the G, N, and M proteins--as shown by immunoprecipitation of [35S]methionine-labeled spheroplast lysates. In NaDodSO4/polyacrylamide gel electrophoresis, M and N proteins comigrated with those of the virion, whereas the yeast-made G protein migrated as two bands with apparent molecular sizes of 60 and 70 kDa. Both polypeptides appeared to be N-glycosylated, since only one polypeptide with the apparent molecular mass of approximately equal to 55 kDa was produced in the presence of tunicamycin. Phase separation into Triton X-114 suggested that the unglycosylated G protein was membrane bound. According to immunofluorescent surface staining of live spheroplasts, at least part of the G protein was transported to the plasma membrane. Spheroplasts expressing the VSV genes could be fused together by low pH to form polykaryons, indicating that G protein synthetized by yeast was fusogenic--i.e., biologically active.


Complex febrile seizures associated with influenza A.


title: Virus profile in children with acute respiratory infections with various severities in Beijing, China.

text: BACKGROUND Acute respiratory infection (ARI) is one of the most common infectious diseases in infants and young children globally. This study aimed to determine the virus profile in children with ARI presenting with different severities. METHODS Clinical specimens collected from children with ARI in Beijing from September 2010 to March 2011 were investigated for 18 respiratory viruses using an xTAG Respiratory Viral Panel Fast (RVP Fast) assay. The Pearson chi-square analysis was used to identify statistical significance. RESULTS Of 270 cases from three groups of ARI patients, including Out-patients, In-patients and patients in the intensive care unit (ICU), viruses were detected in 176 (65.2%) specimens with the RVP Fast assay. The viral detection rate from the Out-patients group (50.0%) was significantly lower than that from the In-patients (71.1%) and ICU-patients (74.4%) groups. The virus distribution was different between the Out-patients group and the other hospitalized groups, while the virus detection rate and distribution characteristics were similar between the In-patients and ICU-patients groups. The co-infection rates of the Out-patients group, the In-patients group, and the ICU-patients group were 15.6%, 50.0% and 35.8%, respectively. In addition to respiratory syncytial virus (RSV) and adenovirus (ADV), human rhinovirus (HRV) was frequently detected from children with serious illnesses, followed by human metapneumovirus (hMPV), human bocavirus (HBoV) and coronaviruses. Parainfluenza virus 3 (PIV3) was detected in children with lower respiratory illness, but rarely from those with serious illnesses in the ICU-patient group. CONCLUSION In addition to so-called common respiratory viruses, virus detection in children with ARI should include those thought to be uncommon respiratory viruses, especially when there are severe ARI-related clinical illnesses.


title: Viral etiology of acute respiratory tract infections in children presenting to hospital: role of polymerase chain reaction and demonstration of multiple infections.

text: BACKGROUND Viral lower respiratory tract infections are a leading cause of hospitalization for young children. METHODS We used polymerase chain reaction (PCR) and conventional methods of cell culture and antigen detection to establish the viral etiology of acute respiratory tract infections in 75 hospitalized children. RESULTS One or more viral pathogens were detected in 65 (87%) children, with respiratory syncytial virus being the most commonly identified virus (36 children). Other viruses identified included influenza virus types A and B, parainfluenzavirus type 3, adenovirus, enterovirus, rhinovirus, coronavirus and human metapneumovirus. PCR increased the diagnostic yield significantly compared with antigen detection and culture, with 39 (21%) diagnoses identified by this method. Multiple infections were identified in 20 (27%) children. CONCLUSIONS PCR-based methodologies offer increased sensitivity for the detection of most respiratory viruses in young children. The inclusion of PCR into diagnostic testing strategies is needed to broaden our understanding of the natural ecology of respiratory viruses and the significance of multiple infections.


title: Strategy for systematic assembly of large RNA and DNA genomes: transmissible gastroenteritis virus model.

text: A systematic method was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in nature. The approximately 30-kb genome, coupled with regions of genomic instability, has hindered the development of a full-length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine. Using a novel approach, six adjoining cDNA subclones that span the entire TGEV genome were isolated. Each clone was engineered with unique flanking interconnecting junctions which determine a precise systematic assembly with only the adjacent cDNA subclones, resulting in an intact TGEV cDNA construct of approximately 28.5 kb in length. Transcripts derived from the full-length TGEV construct were infectious, and progeny virions were serially passaged in permissive host cells. Viral antigen production and subgenomic mRNA synthesis were evident during infection and throughout passage. Plaque-purified virus derived from the infectious construct replicated efficiently and displayed similar plaque morphology in permissive host cells. Host range phenotypes of the molecularly cloned and wild-type viruses were similar in cells of swine and feline origin. The recombinant viruses were sequenced across the unique interconnecting junctions, conclusively demonstrating the marker mutations and restriction sites that were engineered into the component clones. Full-length infectious constructs of TGEV will permit the precise genetic modification of the coronavirus genome. The method that we have designed to generate an infectious cDNA construct of TGEV could theoretically be used to precisely reconstruct microbial or eukaryotic genomes approaching several million base pairs in length.


title: Replication-defective bovine adenovirus type 3 as an expression vector.

text: Although recombinant human adenovirus (HAV)-based vectors offer several advantages for somatic gene therapy and vaccination over other viral vectors, it would be desirable to develop alternative vectors with prolonged expression and decreased toxicity. Toward this objective, a replication-defective bovine adenovirus type 3 (BAV-3) was developed as an expression vector. Bovine cell lines designated VIDO R2 (HAV-5 E1A/B-transformed fetal bovine retina cell [FBRC] line) and 6.93.9 (Madin-Darby bovine kidney [MDBK] cell line expressing E1 proteins) were developed and found to complement the E1A deletion in BAV-3. Replication-defective BAV-3 with a 1.7-kb deletion removing most of the E1A and E3 regions was constructed. This virus could be grown in VIDO R2 or 6.93.9 cells but not in FBRC or MDBK cells. The results demonstrated that the E1 region of HAV-5 has the capacity to transform bovine retina cells and that the E1A region of HAV-5 can complement that of BAV-3. A replication-defective BAV-3 vector expressing bovine herpesvirus type 1 glycoprotein D from the E1A region was made. A similar replication-defective vector expressing the hemagglutinin-esterase gene of bovine coronavirus from the E3 region was isolated. Although these viruses grew less efficiently than the replication-competent recombinant BAV-3 (E3 deleted), they are suitable for detailed studies with animals to evaluate the safety, duration of foreign gene expression, and ability to induce immune responses. In addition, a replication-competent recombinant BAV-3 expressing green fluorescent protein was constructed and used to evaluate the host range of BAV-3 under cell culture conditions. The development of bovine E1A-complementing cell lines and the generation of replication-defective BAV-3 vectors is a major technical advancement for defining the use of BAV-3 as vector for vaccination against diseases of cattle and somatic gene therapy in humans.


title: Non Susceptibility of Neonatal and Adult Rats against the Middle East Respiratory Syndrome Coronavirus.

text: The present study examined the susceptibility of rats to the Middle East respiratory syndrome coronavirus (MERS-CoV) and determined whether this animal is a suitable model for MERS-CoV infection. Immunohistochemical analysis identified dipeptidyl peptidase 4 (DPP4), a known receptor for MERS-CoV on type I pneumocytes from infected rats. Whereas adult rats developed antibodies against MERS-CoV spike protein after intranasal inoculation, there was no evidence of viral replication in the lungs of adult, young, or neonatal rats after intranasal inoculation with MERS-CoV. In addition, human DPP4-expressing rat kidney fibroblasts, but not rat DPP4-expressing cells, were susceptible to MERS-CoV. Taken together, these results suggest that the rat is not a useful animal model for studying MERS-CoV infection.


title: Discovery of novel human and animal cells infected by the severe acute respiratory syndrome coronavirus by replication-specific multiplex reverse transcription-PCR.

text: The severe acute respiratory syndrome coronavirus (SARS-CoV) is the causative agent of the recent outbreak of severe acute respiratory syndrome. VeroE6 cells, fetal rhesus monkey kidney cells, and human peripheral blood mononuclear cells were the only cells known to be susceptible to SARS-CoV. We developed a multiplex reverse transcription-PCR assay to analyze the susceptibility of cells derived from a variety of tissues and species to SARS-CoV. Additionally, productive infection was determined by titration of cellular supernatants. Cells derived from three species of monkey were susceptible to SARS-CoV. However, the levels of SARS-CoV produced differed by 4 log(10). Mink lung epithelial cells (Mv1Lu) and R-Mix, a mixed monolayer of human lung-derived cells (A549) and mink lung-derived cells (Mv1Lu), are used by diagnostic laboratories to detect respiratory viruses (e.g., influenza virus); they were also infected with SARS-CoV, indicating that the practices of diagnostic laboratories should be examined to ensure appropriate biosafety precautions. Mv1Lu cells produce little SARS-CoV compared to that produced by VeroE6 cells, which indicates that they are a safer alternative for SARS-CoV diagnostics. Evaluation of cells permissive to other coronaviruses indicated that these cell types are not infected by SARS-CoV, providing additional evidence that SARS-CoV binds an alternative receptor. Analysis of human cells derived from lung, kidney, liver, and intestine led to the discovery that human cell lines were productively infected by SARS-CoV. This study identifies new cell lines that may be used for SARS-CoV diagnostics and/or basic research. Our data and other in vivo studies indicate that SARS-CoV has a wide host range, suggesting that the cellular receptor(s) utilized by SARS-CoV is highly conserved and is expressed by a variety of tissues.


title: Molecular epizootiology of infectious bronchitis virus in Sweden indicating the involvement of a vaccine strain.

text: To improve the detection and molecular identification of infectious bronchitis virus (avian coronavirus), two reverse transcriptase-polymerase chain reaction (PCR) assays were developed. As 'diagnostic#10; PCR', a set of consensus nested primers was selected from highly conserved stretches of the nucleocapsid (N) gene. As 'phylogeny' PCR, a fragment of the spike protein gene (S1) was amplified and the PCR products were directly sequenced. To study the phylogenetic relationships of the viruses from various outbreaks, studies of molecular epizootiology were performed in Sweden, a Nordic region, where the occurrence of natural cases of the disease is relatively low and the occasional use of live vaccine(s) is well recorded and monitored. The disease appeared in the region in 1994, associated with production problems among layers of various ages. During outbreaks in 1995 and 1997, both layers and broilers were affected. To reduce losses, a live attenuated vaccine has been applied since 1997. By examining 12 cases between 1994 and 1998, molecular epizootiology revealed that, before 1997, the viruses had gene sequences very similar to strains of the Massachusetts serotype. However, comparative sequence analysis of the S1 gene revealed that the identity was not 100% to any of the strains of this serotype that we analysed. A virus related to the Dutch-type strain, D274, was also identified on one farm. Surprisingly, from 1997, the year that vaccination commenced with a live Massachusetts serotype vaccine, the majority of viruses detected had S1 sequences identical to the live Massachusetts vaccine strain. This genetic relation to the vaccine virus was also confirmed by N gene sequence analysis. The studies of molecular epizootiology reveal a strong probability that the vaccination had lead to the spread of the vaccine virus, causing various disease manifestations and a confusing epizootiological situation in the poultry population.


title: Global Health Workforce Alliance: increasing the momentum for health workforce development.

text: The Global Health Workforce Alliance was launched in 2006 to provide a joint platform for governments, development partners, international agencies, civil society organizations, academia, private sector, professional associations, and other stakeholders to work together to address a global crisis in human resources for health. Five years later the vision and mandate of the Alliance still remain valid. Despite advances in bringing the health workforce to the fore in international health policy arenas, more available knowledge and tools, and encouraging signs of commitments from many countries, health workforce bottlenecks continue to prevent many health systems from delivering essential and quality health services. Latin America is not spared from the challenges. The 2010 Second Global Forum on Human Resources for Health provided an opportunity to review progress, identify persisting gaps, reach consensus on solutions, and renew the momentum for and commitment to acutely needed investment and actions.


title: The 5' end of coronavirus minus-strand RNAs contains a short poly(U) tract.

text: A radiolabeled oligodeoxynucleotide primer that anneals near the common 5' end of bovine coronavirus minus-strand RNAs was extended with reverse transcriptase, and a major product suggesting poly(U) tracts of 8 to 20 nucleotides was found. The extended primer molecules were ligated head to tail, amplified by the polymerase chain reaction, cloned, and sequenced, and poly(U) tracts of 9 to 26 nucleotides were found. Poly(A) tails of 100 to 130 nucleotides on the 3' end of coronavirus plus-strand mRNAs and genome must, therefore, be generated by a mechanism that uses only a short poly(U) template. This pattern contrasts with that of other cytoplasmic, polyadenylated, plus-strand animal RNA viruses which utilize a full-length poly(U) template for poly(A) synthesis.


title: Renal involvement in patients with hepatitis C virus infection.

text: INTRODUCTION Hepatitis C virus (HCV) infection is a hepatotropic virus causing a variety of extrahepatic immunological manifestations and is a risk factor of a variety of extrahepatic diseases, such as mixed cryoglobulinemia and membranoproliferative glomerulonephritis (MPGN), which is the most common glomerulonephritis. The aim of this study was to evaluate renal involvement in HCV-infected patients. MATERIALS AND METHODS A total of 300 randomly-selected HCV antibody-positive outpatients at the HCV clinic of Shariati hospital were enrolled. Serum creatinine was measured and glomerular filtration rate was estimated accordingly. Urine proteinuria was measured in 24-hour urine samples. RESULTS The patients were 249 men (83.2%) and 51 women (16.8%) with a mean age of 37.8 +/- 11.7 years (range, 18 to 70 years). Proteinuria was found in 12 HCV antibody-positive adults (4%), 1 of whom underwent biopsy. He was a 55- year-old man with a 4-month history of facial and lower extremities edema and 3-g proteinuria with a normal kidney function (glomerular filtration rate, 85 mL/min) and normocomplementemia. Kidney biopsy specimens showed MPGN. The frequency of low glomerular filtration rate was 0.7% (2 patients) in the HCV antibody-positive adults. There was no significant relationship between HCV seropositivity and low glomerular filtration rate. CONCLUSIONS Our observations showed renal involvement in HCV antibody-positive patients. Among immune complex glomerular kidney diseases, MPGN without cryoglobulins is thought to be the most common in these patients.


title: Psychological distress of nurses in Taiwan who worked during the outbreak of SARS.

text: OBJECTIVES This study explored whether nurses who were working during the severe acute respiratory syndrome (SARS) crisis showed symptoms of distress. This study also examined whether nurses experienced stress from being conscripted to work in higher-risk units. METHODS This study was conducted in Taiwan during the peak of the SARS outbreak in 2003. Participants were recruited from nursing staff in a community hospital. Participants were divided into three groups: those who originally worked in units with a high risk of SARS exposure (N=65), those who had been involuntarily conscripted into such units because of manpower demands (N=21), and those who worked in units with a low risk of SARS exposure (N=45). Participants were evaluated with two questionnaires: the Impact of Event Scale and the 90-item Symptom Checklist-Revised. RESULTS The results showed that 11 percent of the nurses surveyed had stress reaction syndrome. The symptoms of psychological stress reactions included anxiety, depression, hostility, and somatization. The highest rate of stress reaction syndrome was observed in the group that originally worked in a high-risk unit, and the conscripted group experienced the most severe distress on average. CONCLUSIONS These findings suggest that the psychological distress of nurses who worked during the SARS outbreak was moderate. The distress was more prominent among the two groups of nurses who were working with patients with SARS.


title: A simple approximate mathematical model to predict the number of severe acute respiratory syndrome cases and deaths.

text: BACKGROUND Severe acute respiratory syndrome (SARS) is currently spreading in many countries. This paper proposes a simple approximate mathematical model for public health practitioners to predict the number of SARS cases and deaths. METHODS The model is based on four parameters: R(o) (basic reproductive number), F (case-fatality rate), i (incubation period), and d (duration of disease). The calculations can be done by hand or by using a computer spreadsheet. RESULTS The best parameters to fit Canadian data as of 6 April 2003 (before infection controls took effect) are R(o) = 1.5, F = 30%, i = 5 days, d = 14 days. On 6 April (day 40) there were 74 cases and 7 deaths. If this trend continues, SARS numbers in Canada are predicted to be as follows: 387 cases and 34 deaths by 26 April (day 60), 4432 cases and 394 deaths by 26 May (day 90), and 50 500 cases and 4489 deaths by 25 June (day 120). By comparison, the best parameters to fit Hong Kong data as of 10 April 2003 are R(o) = 2.0, F = 20%, i = 5 days, d = 14 days. CONCLUSIONS Using the proposed mathematical model, it was estimated that about 1.5 to 2 new infectious cases were produced per infectious case every five days. Also, about 20% to 30% of the cases die within 14 days. The case-fatality may therefore be considerably higher than initially thought. The model indicates that SARS can spread very fast when there are no interventions.


title: Soluble receptor potentiates receptor-independent infection by murine coronavirus.

text: Mouse hepatitis virus (MHV) infection spreads from MHV-infected DBT cells, which express the MHV receptor CEACAM1 (MHVR), to BHK cells, which are devoid of the receptor, by intercellular membrane fusion (MHVR-independent fusion). This mode of infection is a property of wild-type (wt) JHMV cl-2 virus but is not seen in cultures infected with the mutant virus JHMV srr7. In this study, we show that soluble MHVR (soMHVR) potentiates MHVR-independent fusion in JHMV srr7-infected cultures. Thus, in the presence of soMHVR, JHMV srr7-infected DBT cells overlaid onto BHK cells induce BHK cell syncytia and the spread of JHMV srr7 infection. This does not occur in the absence of soMHVR. soMHVR also enhanced wt virus MHVR-independent fusion. These effects were dependent on the concentration of soMHVR in the culture and were specifically blocked by the anti-MHVR monoclonal antibody CC1. Together with these observations, direct binding of soMHVR to the virus spike (S) glycoprotein as revealed by coimmunoprecipitation demonstrated that the effect is mediated by the binding of soMHVR to the S protein. Furthermore, fusion of BHK cells expressing the JHMV srr7 S protein was also induced by soMHVR. These results indicated that the binding of soMHVR to the S protein expressed on the DBT cell surface potentiates the fusion of MHV-infected DBT cells with nonpermissive BHK cells. We conclude that the binding of soMHVR to the S protein converts the S protein to a fusion-active form competent to mediate cell-cell fusion, in a fashion similar to the fusion of virus and cell membranes.


title: Prevalence of antibodies against transmissible gastroenteritis virus and porcine respiratory coronavirus among pigs in six regions in Japan.

text: A total of 2,703 pig sera from 171 farms in six regions in Japan were screened for virus-neutralizing (VN) antibody against transmissible gastroenteritis virus (TGEV). Although none of the farms had clinical signs of transmissible gastroenteritis (TGE) or vaccination against TGEV, VN antibody was detected in 14.4% of sera at 30 farms (17.5%) across all six regions. On testing of 263 VN antibody-positive sera from 27 farms with a commercial blocking ELISA to distinguish TGEV and porcine respiratory coronavirus (PRCV) antibodies, 78.3% were positive for PRCV antibody only, while 12.5% were positive for TGEV antibody only or both TGEV and PRCV antibodies. Seven of the eight TGEV antibody-positive farms were also positive for PRCV antibody. Five months after the antibody examination, a TGE outbreak occurred at one of these seven farms. These results suggest that most of the detected VN antibodies were to PRCV, and that TGEV infection might be present at some PRCV-positive farms in Japan.


title: Olfactory threshold and nasal mucosal changes in experimentally induced common cold.

text: Common cold is a most common disease in man accompanied by an experience of a diminished sense of smell. Controlled studies of the sense of smell are lacking and the cause as well as degree of impairment of smell in common cold has not been satisfactorily evaluated. We have investigated whether impaired olfactory ability accompanies common cold and if the loss is related to nasal blockage and to the amount of nasal discharge. For this purpose we measured the threshold of olfaction in volunteers before and after nasal inoculation with coronavirus 229E. The absolute olfactory threshold was assessed by a modified discrimination step test with dilutions of butanol. Symptoms of common cold were registered using a four-grade scale. Nasal obstruction was measured by nasal peak expiratory flow and by acoustic rhinometry and nasal discharge by weight of pre-weighted handkerchiefs. Individuals with a cold had impaired olfaction and the change in smelling ability correlated to the nasal congestion but not to the nasal discharge in analysis of multiple linear regression.


title: The link between environmental toxicant exposure and endometriosis.

text: Endometriosis is an estrogen-dependent disease characterized by the growth of endometrial cells in ectopic locations. Although the etiology of endometriosis is unknown, several hypotheses have been proposed to explain its origin. Retrograde menstruation of endometrial cells into the peritoneum is the most widely accepted theory, however, this phenomenon occurs in approximately 90% of women while the prevalence of endometriosis is much lower. Hence, other factors are thought to contribute to the development of this disease, including exposure to environmental toxicants. Although the epidemiological evidence is equivocal, animal and experimental investigations provide a basis for the proposed association between dioxin and dioxin-like chemical exposure and endometriosis. However, the mechanism(s) underlying this potential association are poorly understood. Development of novel animal models that more reliably recapitulate the pathogenesis and pathophysiology of this disease provide exciting opportunities to further test the link between exposure to these chemicals and endometriosis. Moreover, differential expression of several novel genes that may be important in the disease provides new targets to test the actions of environmental toxicants in the pathobiology of endometriosis.


title: Factors associated with development of Canine Infectious Respiratory Disease Complex (CIRDC) in dogs in 5 Canadian small animal clinics.

text: This study investigated the association between presence of respiratory pathogens and development of Canine Infectious Respiratory Disease Complex (CIRDC) in dogs in 5 Canadian small animal clinics. In total, 86 dogs were tested using a commercial PCR respiratory panel; 64 dogs were considered as cases and 22 were control dogs matched by veterinary clinic. No control animals (0/22) were positive for canine parainfluenza virus (CPIV), whereas 27/64 (42%) CIRDC cases were positive. Furthermore, 81% of case dogs tested positive for Mycoplasma cynos, compared with 73% of control dogs. Canine respiratory corona virus (CRCoV) was detected in no control dogs compared with 9.4% of clinical dogs. No animals were positive for any influenza virus type A present in the diagnostic panel. Presence of CPIV was associated (P < 0.01) with the occurrence of CIRDC after adjustment for demographic factors and presence of CRCoV (P = 0.09).


title: The Baycrest SARS experience: the human side.

text: Toronto, in the province of Ontario, Canada was one of the cities severely impacted by Severe Acute Respiratory Syndrome (SARS). SARS required the health care system to respond quickly and efficiently. This paper describes the situation and response at a large public academic aged care centre.


Sex, SARS, and the Holy Grail.


title: Characterization of an enteropathogenic bovine calicivirus representing a potentially new calicivirus genus.

text: Bovine enteric caliciviruses (BEC) are associated with diarrhea in young calves. The BEC strains detected in Europe form a third genogroup within the genus "Norwalk-like viruses" (NLV) of the family Caliciviridae. In this report, we present sequence, clinical, and histological data characterizing a novel enteropathogenic BEC strain, NB, detected in fecal specimens from calves in the United States. The complete RNA genome of the NB virus is 7,453 bases long and is organized into two open reading frames (ORFs). ORF-1 is 2,210 amino acids long and encodes a large nonstructural polyprotein contiguous with the major capsid protein (VP1), similar to the lagoviruses and "Sapporo-like viruses" (SLV). The conserved calicivirus motifs were identified in the nonstructural proteins. ORF-2 is located at the 3' end of the genome and encodes a small basic protein (VP2) of 225 amino acids. The 5' and 3' untranslated regions are 74 and 67 bases long, respectively. Among caliciviruses, NB virus shows amino acid identities of 14.1 to 22.6% over the entire ORF-1 nonstructural-protein sequence with NLV, SLV, vesivirus, and lagovirus strains, while the overall sequence identity of the complete NB VP-1 with other caliciviruses is low, varying between 14.6 and 26.7%. Phylogenetic analysis of the complete VP1 protein, including strains from all four calicivirus genera, showed the closest grouping of NB virus to be with viruses in the genus Lagovirus, which cause liver infections and systemic hemorrhage in rabbits. In gnotobiotic calves, however, NB virus elicited only diarrhea and intestinal lesions that were most severe in the upper small intestine (duodenum and jejunum), similar to the NLV BEC strains. The tissues of major organs, including the lung, liver, kidney, and spleen, had no visible microscopic lesions.


title: Modulation of in vivo immune response by selective depletion of neutrophils using a monoclonal antibody, RP-3. II. Inhibition by RP-3 treatment of mononuclear leukocyte recruitment in delayed-type hypersensitivity to sheep red blood cells in rats.

text: We demonstrated in our previous paper that treatment of rats with RP-3, a mAb that depletes in vivo neutrophils selectively, resulted in inhibition of both the priming and effector phases of delayed type hypersensitivity (DTH) to SRBC. In order to clarify the mechanisms of this phenomenon, we examined the effect of RP-3 treatment on mononuclear leukocyte (MNL) recruitment in DTH. When rats had been treated with RP-3 at the time of Ag priming, MNL migration, which accompanies DTH, was inhibited. The prior infiltration of neutrophils was also partially required for MNL recruitment because migration was inhibited when the immune rats were treated with RP-3 at the time of DTH elicitation.