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.


[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.


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.


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.


title: Characterization of and functional antigen presentation by central nervous system mononuclear cells from mice infected with Theiler's murine encephalomyelitis virus.

text: We examined the phenotype and function of cells infiltrating the central nervous system (CNS) of mice persistently infected with Theiler's murine encephalomyelitis virus (TMEV) for evidence that viral antigens are presented to T cells within the CNS. Expression of major histocompatibility complex (MHC) class II in the spinal cords of mice infected with TMEV was found predominantly on macrophages in demyelinating lesions. The distribution of I-As staining overlapped that of the macrophage marker sialoadhesin in frozen sections and coincided with that of another macrophage/microglial cell marker, F4/80, by flow cytometry. In contrast, astrocytes, identified by staining with glial fibrillary acidic protein, rarely expressed detectable MHC class II, although fibrillary gliosis associated with the CNS damage was clearly seen. The costimulatory molecules B7-1 and B7-2 were expressed on the surface of most MHC class II-positive cells in the CNS, at levels exceeding those found in the spleens of the infected mice. Immunohistochemistry revealed that B7-1 and B7-2 colocalized on large F4/80(+) macrophages/microglia in the spinal cord lesions. In contrast, CD4(+) T cells in the lesions expressed mainly B7-2, which was found primarily on blastoid CD4(+) T cells located toward the periphery of the lesions. Most interestingly, plastic-adherent cells freshly isolated from the spinal cords of TMEV-infected mice were able to process and present TMEV and horse myoglobin to antigen-specific T-cell lines. Furthermore, these cells were able to activate a TMEV epitope-specific T-cell line in the absence of added antigen, providing conclusive evidence for the endogenous processing and presentation of virus epitopes within the CNS of persistently infected SJL/J mice.


title: Sacral mesh pullout strength based on suture location for sacrocolpopexy.

text: BACKGROUND We sought to compare the pullout strengths of the sacral end of the sacrocolpopexy mesh when attached using one suture at the first versus second sacral vertebral level (S1 vs. S2) in female cadaveric pelvises. METHODS The sacral vertebrae were isolated in 9 unembalmed female cadavers, and segments of polypropylene mesh were attached to the sacrum using stitches at either the S1 or S2 level. The free end of the mesh was pulled at a constant rate. Maximum tension prior to system failure was recorded for each specimen. Suture pullout strengths between the two groups were compared using the two-sample Wilcoxon rank-sum test. RESULTS In all but one specimen, failure of the system occurred at the point of suture attachment to the anterior longitudinal ligament. Among the nine specimens, median pullout strengths were 7.49 lb (interquartile range 7.95) at S1 and 3.15 lb (interquartile range 2.975) at S2 (p = 0.028). CONCLUSION The pullout strength of sutures used to attach the sacrocolpopexy mesh to the sacrum is significantly higher at the S1 level than at the S2 level.


title: Severe acute respiratory syndrome in a medical center in Taipei.

text: From April 18 to May 31, 2003, 46 patients with probable severe acute respiratory syndrome were admitted to the negative-pressure isolation rooms of Mackay Memorial Hospital in Taipei, Taiwan. Their demographic, clinical, laboratory, and radiologic characteristics and clinical outcomes were analyzed. There were 15 males and 31 females, in this cohort, 13 of whom were healthcare workers. The latter included 6 hospital staff and 7 medical personnel transferred from other hospitals. The most common symptoms were fever (100%, 46/46), cough (72%, 33/46), shortness of breath (46%, 21/46), and diarrhea (39%, 18/46). Other common findings were lymphopenia (57%, 26/46), thrombocytopenia (39%, 18/46), elevated lactate dehydrogenase (63%, 29/46), and elevated creatine kinase (24%, 11/46). A total of 7 patients (15%) required mechanical ventilation, and 8 (17%) died. Advanced age was an independent significant risk factor for death. Fever followed by rapidly progressive respiratory compromise led to significant morbidity and mortality in this cohort.


title: Severe acute respiratory syndrome (SARS): a year in review.

text: Severe acute respiratory syndrome (SARS) emerged from China as an untreatable and rapidly spreading respiratory illness of unknown etiology. Following point source exposure in February 2003, more than a dozen guests infected at a Hong Kong hotel seeded multi-country outbreaks that persisted through the spring of 2003. The World Health Organization responded by invoking traditional public health measures and advanced technologies to control the illness and contain the cause. A novel coronavirus was implicated and its entire genome was sequenced by mid-April 2003. The urgency of responding to this threat focused scientific endeavor and stimulated global collaboration. Through real-time application of accumulating knowledge, the world proved capable of arresting the first pandemic threat of the twenty-first century, despite early respiratory-borne spread and global susceptibility. This review synthesizes lessons learned from this remarkable achievement. These lessons can be applied to re-emergence of SARS or to the next pandemic threat to arise.


Tetanus in injecting drug users, United Kingdom.


title: A statistical analysis protocol for the time-differentiated target temperature management after out-of-hospital cardiac arrest (TTH48) clinical trial.

text: BACKGROUND The TTH48 trial aims to determine whether prolonged duration (48 hours) of targeted temperature management (TTM) at 33 (±1) °C results in better neurological outcomes compared to standard duration (24 hours) after six months in comatose out-of-hospital cardiac arrest (OHCA) patients. METHODS TTH48 is an investigator-initiated, multicentre, assessor-blinded, randomised, controlled superiority trial of 24 and 48 hours of TTM at 33 (±1) ° C performed in 355 comatose OHCA patients aged 18 to 80 years who were admitted to ten intensive care units (ICUs) in six Northern European countries. The primary outcome of the study is the Cerebral Performance Category (CPC) score observed at six months after cardiac arrest. CPC scores of 1 and 2 are defined as good neurological outcomes, and CPC scores of 3, 4 and 5 are defined as poor neurological outcomes. The secondary outcomes are as follows: mortality within six months after cardiac arrest, CPC at hospital discharge, Glasgow Coma Scale (GCS) score on day 4, length of stay in ICU and at hospital and the presence of any adverse events such as cerebral, circulatory, respiratory, gastrointestinal, renal, metabolic measures, infection or bleeding. With the planned sample size, we have 80% power to detect a 15% improvement in good neurological outcomes at a two-sided statistical significance level of 5%. DISCUSSION We present a detailed statistical analysis protocol (SAP) that specifies how primary and secondary outcomes should be evaluated. We also predetermine covariates for adjusted analyses and pre-specify sub-groups for sensitivity analyses. This pre-planned SAP will reduce analysis bias and add validity to the findings of this trial on the effect of length of TTM on important clinical outcomes after cardiac arrest. TRIAL REGISTRATION ClinicalTrials.gov: NCT01689077 , 17 September 2012.


title: Advances in detecting and responding to threats from bioterrorism and emerging infectious disease.

text: Much progress has been made in recent years to strengthen local, state, national and international capacities to detect and respond to bioterrorism events and naturally occurring outbreaks of disease. New tools and systems are available to estimate the potential impact of a biological event and predict resource needs for effective response, enable earlier detection of an attack or outbreak, enhance diagnostic capacity and facilitate rapid intervention to mitigate the impact of an event on a community. These advances have required new approaches to preparedness, planning and surveillance, as well as new partnerships and collaborations across a range of disciplines. We examine some of these developments, discuss potential uses and limitations of these approaches, and identify priorities for the future.


title: Targeted recombination demonstrates that the spike gene of transmissible gastroenteritis coronavirus is a determinant of its enteric tropism and virulence.

text: Targeted recombination within the S (spike) gene of transmissible gastroenteritis coronavirus (TGEV) was promoted by passage of helper respiratory virus isolates in cells transfected with a TGEV-derived defective minigenome carrying the S gene from an enteric isolate. The minigenome was efficiently replicated in trans and packaged by the helper virus, leading to the formation of true recombinant and pseudorecombinant viruses containing the S proteins of both enteric and respiratory TGEV strains in their envelopes. The recombinants acquired an enteric tropism, and their analysis showed that they were generated by homologous recombination that implied a double crossover in the S gene resulting in replacement of most of the respiratory, attenuated strain S gene (nucleotides 96 to 3700) by the S gene of the enteric, virulent isolate. The recombinant virus was virulent and rapidly evolved in swine testis cells by the introduction of point mutations and in-phase codon deletions in a domain of the S gene (nucleotides 217 to 665) previously implicated in the tropism of TGEV. The helper virus, with an original respiratory tropism, was also found in the enteric tract, probably because pseudorecombinant viruses carrying the spike proteins from the respiratory strain and the enteric virus in their envelopes were formed. These results demonstrated that a change in the tropism and virulence of TGEV can be engineered by sequence changes in the S gene.


title: Laparoscopic minimally invasive total gastrectomy with linear stapled oesophagojejunostomy--experience from the first thirty procedures.

text: BACKGROUND There are only few reports on total gastrectomy by a laparoscopic surgical approach. One explanation is the fear of complications due to anastomotic dehiscence in oesophagojejunal anastomosis known to carry high morbidity and mortality. The introduction of staplers have contributed to making anastomosis safer and easier to perform and has facilitated more advanced laparoscopic surgery. In open surgery, most surgeons use a circular stapler for oesophagojejunal anastomosis or a hand sutured technique. Both techniques are difficult to use in laparoscopic surgery, especially if the oesophagus is narrow. To facilitate the creation of oesophagojejunal anastomoses, we have adopted a technique with a linear stapled anastomosis. Our method is based on a stapling technique where the oesophagus is divided above the gastric cardia followed by a oesophagojejunostomy performed with Covidien's new Endo GIA-60™ Ultra Universal stapler. The residual opening is closed with a 3-0 re-absorbable suture. PATIENTS AND METHODS From June 2009 to May 2012, 14 men and 16 women (median age=66 years, range=39-84 years) underwent laparoscopic total gastrectomy due to gastric cancer. RESULTS One patient died during hospital stay; corresponding to a postoperative mortality of 3,3%. Leakage in the oesophagojejunal anastomosis occurred in three patients (10%). Two of the patients with leakage in the oesophagojejunal anastomosis had an additional duodenal bulb leakage, which might have caused anastomotic dehiscence. The patients had a median postoperative hospital stay of six days (range=3-156 days). Six patients had a re-operation due to complications, including one endoscopic stent application in the anastomosis. CONCLUSION Even though a leakage rate of 10% can be considered high, this study describes a simple method for performing oesophagojejunostomy after gastrectomy by a laparoscopic approach independently of the width of the oesophagus. This study also shows that laparoscopic gastrectomy can be performed in more advanced stages of gastric cancer.


Aetiological role of viruses in multiple sclerosis: a review.


title: Potential subversion of autophagosomal pathway by picornaviruses.

text: The RNA replication complexes of small positive-strand RNA viruses such as poliovirus are known to form on the surfaces of membranous vesicles in the cytoplasm of infected mammalian cells. These membranes resemble cellular autophagosomes in their double-membraned morphology, cytoplasmic lumen, lipid-rich composition and the presence of cellular proteins LAMP 1 and LC3. Furthermore, LC3 protein is covalently modified during poliovirus infection in a manner indistinguishable from that observed during bona fide autophagy. This covalent modification can also be induced by the expression of viral protein 2BC in isolation. However, differences between poliovirus-induced vesicles and autophagosomes also exist: the viral-induced membranes are smaller, at 200-400 nm in diameter, and can be induced by the combination of two viral proteins, termed 2BC and 3A. Experimental suppression of expression of proteins in the autophagy pathway was found to reduce viral yield, arguing that this pathway facilitates viral infection, rather than clearing it. We have hypothesized that, in addition to providing membranous surfaces for assembly of viral RNA replication complexes, double-membraned vesicles provide a topological mechanism to deliver cytoplasmic contents, including mature virus, to the extracellular milieu without lysing the cell.


title: Coronavirus genomic and subgenomic minus-strand RNAs copartition in membrane-protected replication complexes.

text: The majority of porcine transmissible gastroenteritis coronavirus plus-strand RNAs (genome and subgenomic mRNAs), at the time of peak RNA synthesis (5 h postinfection), were not found in membrane-protected complexes in lysates of cells prepared by Dounce homogenization but were found to be susceptible to micrococcal nuclease (85%) or to sediment to a pellet in a cesium chloride gradient (61%). They therefore are probably free molecules in solution or components of easily dissociable complexes. By contrast, the majority of minus-strand RNAs (genome length and subgenomic mRNA length) were found to be resistant to micrococcal nuclease (69%) or to remain suspended in association with membrane-protected complexes following isopycnic sedimentation in a cesium chloride gradient (85%). Furthermore, 35% of the suspended minus strands were in a dense complex (1.20 to 1.24 g/ml) that contained an RNA plus-to-minus-strand molar ratio of approximately 8:1 and viral structural proteins S, M, and N, and 65% were in a light complex (1.15 to 1.17 g/ml) that contained nearly equimolar amounts of plus- and minus-strand RNAs and only trace amounts of proteins M and N. In no instance during fractionation were genome-length minus strands found segregated from sub-genome-length minus strands. These results indicate that all minus-strand species are components of similarly structured membrane-associated replication complexes and support the concept that all are active in the synthesis of plus-strand RNAs.


title: Comparison of intestinal (Illinois strain) and cell culture-adapted (M-HP strain) viral populations of transmissible gastroenteritis of swine.

text: Intestinal and cell culture-adapted viral populations of transmissible gastroenteritis (TGE) of swine were compared by means of sucrose gradient centrifugation, immunnofluorescence, electron microscopy, immune electron microscopy, statistical analysis of the number of plaque-forming units, and ultraviolet sensitivity. Results indicated that the size range and general coronavirus morphologic characteristics were shared by both viral populations. Marked morphologic variations existed among particles from both populations. Unlike the cell culture-adapted virus, the Illinois virus of intestinal origin was infractions representing 2 bands of infectivity which were isolated by the sucrose gradient centrifugation method. The intestinal and cell culture-adapted TGE viruses were similar in antigenicity and in sensitivity to ultraviolet irradiation. There was no indication of a 2nd virus in addition to the coronavirus described as the cause of TGE.


title: Cerebral oximetry to reduce perioperative morbidity.

text: The use of cerebral oximetry to guide intraoperative management has been shown to improve patient outcomes in cardiac surgery. This pilot trial assessed the feasibility of performing a similar study of outcome in patients over the age of 70 years undergoing non-cardiac surgery. Patients over the age of 70 years undergoing total knee or hip arthroplasty or bowel resection surgery were randomly assigned to have cerebral oximetry values monitored (intervention group) or not monitored (control) while under general anaesthesia. Indicators of proof of concept were: rate of complications, rate of cerebral desaturation, relationship between cerebral desaturation and complications, and anaesthetist response to cerebral desaturation. Forty patients were recruited and randomised to a control group (n=20) or an intervention group (n=20). The proportion of the study population who had a complication was 40% (95% confidence interval [CI] 26% to 55%). Cerebral desaturation (<75% of baseline) occurred in only two patients (5.0% (CI 1.4% to 16%)), one in each group, and neither of those patients recorded a complication. Changes to anaesthetic management on the basis of cerebral oximetry values occurred in only two patients in the intervention group (10% (CI 2.8% to 30%)). Maintenance of cerebral oximetry values appeared to be closer to baseline in the intervention group than in the control group but this difference was not significant (P=0.15). Our results indicated that complications occurred frequently in the study population but did not appear to be associated with cerebral desaturation events. These findings do not support a larger intervention study using the current study population.


How disease stage impacts the rate of second primary malignancies in patients with diffuse large B-cell lymphoma.


title: The impact of severe acute respiratory syndrome on medical house staff: a qualitative study.

text: OBJECTIVE To explore the impact of severe acute respiratory syndrome (SARS) on a medical training program and to develop principles for professional training programs to consider in dealing with future, similar crises. DESIGN Qualitative interviews analyzed using grounded theory methodology. SETTING University-affiliated hospitals in Toronto, Canada during the SARS outbreak in 2003. PARTICIPANTS Medical house staff who were allocated to a general internal medicine clinical teaching unit, infectious diseases consultation service, or intensive care unit. RESULTS Seventeen medical residents participated in this study. Participants described their experiences during the outbreak and highlighted several themes including concerns about their personal safety and about the negative impact of the outbreak on patient care, house staff education, and their emotional well-being. CONCLUSION The ability of residents to cope with the stress of the SARS outbreak was enhanced by the communication of relevant information and by the leadership of their supervisors and infection control officers. It is hoped that training programs for health care professionals will be able to implement these tenets of crisis management as they develop strategies for dealing with future health threats.


title: Age-dependent resistance to transmissible gastroenteritis of swine. III. Effects of epithelial cell kinetics on coronavirus production and on atrophy of intestinal villi.

text: Coronavirus titers in small intestine, degree of villous atrophy and apparent rates of regeneration of intestinal villi were compared in newborn, 3-week-old and adult pigs for 1 week after they were exposed to the transmissible gastroenteritis virus of swine. The response within the newborn group was homogeneous, resulting in high virus titers, maximal villous atrophy and comparatively slow regeneration. In general, virus titers were lower, villous atrophy was less severe and regeneration more rapid in both older groups than in the newborn pigs. However, the response varied greatly in the older groups. The 3-week-old group was divided into two populations. The major population had low virus titers and developed partial villous atrophy, whereas the minor population had marked villous atrophy and virus titers comparable to those of the newborn pigs. These observations support the hyposthesis that the accelerated replacement of villous epithelium in the small intestine of pigs during the first 3 weeks contributes to the innate age-dependent resistance to transmissible gastroenteritis. The accelerated replacement of villous epithelial cells in older pigs contributes to resistance in two ways. The increased proliferative capacity of crypt epithelium results in a more rapid regeneration of atrophic villi; and the comparatively young villous absorptive cells resulting from accelerated replacement produce less virus per cell than the older ones of the newborn pig.


title: Coronavirus JHM induced demyelinating disease: specific domains on the E2-protein are associated with neurovirulence.

text: Infections of rodents by murine coronaviruses can lead to chronic diseases of the central nervous system. These infections are interesting systems to study mechanisms which could be relevant for the pathogenesis of certain human diseases. One major factor influencing the outcome of infection is related to the virus. To understand the virological basis for neurovirulence we compared JHM-virus isolates with different biological properties. JHM-Wt causes only acute disease, JHM-Ts43 a demyelinating encephalomyelitis and a virus shedded from persistently infected cells (JHM-Pi) is not virulent at all. The spread of these viruses in glial cell cultures reflects their different neurovirulence for animals. The peplomer E2 of these viruses reveals structural and antigenic differences. We characterised the epitopes of E2 with a panel of monoclonal antibodies. Four epitopes are associated with regions important for neutralisation, cell fusion and attachment. More than five epitopes are not related to such functions. Epitopes differ in their location and accessibility on the E2 protein subunits between JHM-Wt, JHM-Ts43 and JHM-Pi. To identify epitopes in regions important for pathogenesis, we performed animal studies with variants selected by monoclonal antibodies. Variants changed in a defined epitope (E2-Ba) induce in Balb/c mice a chronic disease. Variants changed in only one of the other three neutralisation epitopes induce acute disease. These results support and extend the observation that the peplomer protein E2 is a major determinant for virulence and antigenic variability of coronaviruses 1,4,5,6,8,10,17,19,22,23. Increasing evidence had been obtained that certain structural features of this protein are important for the cell tropism of the virus. Furthermore, this protein influences strongly the type and specificity of immune responses against viral and host antigens. The highly advanced knowledge on structure and replication of coronaviruses will be of great value to analyze further mechanisms leading to inflammatory demyelinating diseases associated with a persistent virus infection.


title: Correlation between maternal serum antibodies and protection against bovine rotavirus diarrhea in calves.

text: The correlation between maternal serum antibodies in beef calves at 2 days old and protection against diarrhea induced by natural bovine rotavirus (BRV) infection was examined. Virus neutralizing (VN) antibody titers against BRV in sera from calves that developed diarrhea by BRV infection within 14 days of age (BRV-diarrheal calves) were significantly lower than those from calves that had no diarrhea. In the BRV-diarrheal calves, a positive correlation was found between the VN antibody titers and age of the onset of diarrhea. There were negative correlations between the VN antibody titers and duration of the diarrhea, VN antibody titers and cumulative diarrhea scores, and the VN antibody titers and duration of virus shedding. These results suggest that the VN antibody titers against BRV in newborn calf serum could be an indicator of protection against BRV-induced diarrhea.


title: Molecular characterization of infectious bronchitis virus strains isolated from the enteric contents of Brazilian laying hens and broilers.

text: Infectious bronchitis virus (IBV) is the causative agent of avian infectious bronchitis, which is characterized by respiratory, reproductive, and renal signs. However, the role of IBV as an enteric pathogen in still controversial. In Brazil, antigenic groups of IBV divergent from the Massachusetts serotype used for vaccination schedules in that country have already been demonstrated. The present study aimed to assess the different genotypes of IBV in Brazilian commercial poultry flocks by partial sequencing of the S1 amino-terminus coding region using enteric contents as samples and examine their relationship with the vaccine serotype currently in use. Samples of enteric contents were taken as pools of five birds from each of 18 poultry farms (17 broiler and one laying farm) from five Brazilian states between 2002 and 2006. Birds were presenting watery diarrhea and poor general condition but were without respiratory, renal, or reproductive signs. Conventional antibacterial and anticoccidial therapies were unsuccessful and, furthermore, all samples proved negative for rotavirus, reovirus, and astrovirus. Eleven IBV samples were isolated in embryonated eggs and resulted in S1 sequences. Phylogenetic analysis showed that these segregated into an exclusive cluster, close to serotype D274, but distant from Massachusetts. Mean amino acid identity amongst these Brazilian strains was 94.07%; amongst these and serotypes D274, 4/91, and Massachusetts, mean amino acid identity was 77.17%, 69.94%, and 68.93%, respectively. In conclusion, the presence of genotype variant strains of IBV in Brazilian poultry flocks has been demonstrated and might be the reason for the unsuccessful control of IBV in Brazil. Furthermore, these results also strengthen the implications of IBV in enteric diseases of poultry.


Dermatophilosis in captive tortoises.


title: Dissemination of MHV4 (strain JHM) infection does not require specific coronavirus receptors.

text: In this report, we demonstrate the syncytial spread of MHV4 (strain JHM) infection through non-murine cell cultures which lack a specific MHV4 receptor and are therefore resistant to infection by free virions. This was achieved by allowing infected murine cells to settle onto confluent monolayers of non-murine cells in a straightforward infectious center assay. Receptor-independent syncytium formation induced by cells expressing the MHV4 spike (S) from recombinant vaccinia viruses (VV) indicated that spread was mediated by this coronavirus glycoprotein. We conclude that the S protein of MHV4 is so potently fusogenic that it does not require prior binding to a virus-specific surface receptor to induce fusion of closely-opposed plasma membranes.