title: Adolescent twin sisters with severe acute respiratory syndrome (SARS).

text: A novel coronavirus-associated communicable respiratory disease, severe acute respiratory syndrome (SARS), spread worldwide after an outbreak in Guangdong Province of the People's Republic of China in November 2002. Since late February 2003, there has been an epidemic in Hong Kong involving both adult and pediatric patients. The clinical course, intensive care, and outcome of adolescent twin sisters with SARS are described. Adolescents infected with SARS may develop severe illness as adults, and close monitoring for disease progression in terms of both clinical and radiologic deterioration is warranted.


title: Rerouting the traffic from a virus perspective.

text: Viruses are important human and animal pathogens causing disease that affect global health and the economy. One outcome of many virus infections is the regulation of cellular trafficking machinery. Viral proteins recruit and interact with cellular trafficking proteins to divert the normal trafficking of key proteins or to induce the formation of novel membrane structures in the host cell. These alterations often increase replication efficiency by mislocalizing immune regulators or restriction factors ot by creating platforms for replication and assembly of new virus particles. Our knowledge of how viruses interact with the cellular trafficking machinery is still limited and furthering this understanding will be important for the future development of prophylactic and therapeutic treatments. This review provides a glimpse of the types of interplay between viral and cellular factors that result in a disruption of cellular trafficking or modifications to cellular membranes.


title: Coexpression and association of the spike protein and the membrane protein of mouse hepatitis virus.

text: The M and S envelope glycoproteins of mouse hepatitis virus associate in the process of virus assembly. We have studied the intrinsic properties of M/S heterocomplexes by coexpressing M and S in the absence of other coronaviral proteins. The formation of M/S complexes under these conditions indicates that M and S can interact independently of other coronaviral factors. Pulse-chase analysis revealed that M and S associate in a pre-Golgi compartment. M/S complexes are efficiently transported beyond the coronavirus budding compartment to the Golgi complex. The failure to detect complexes at the surface of coexpressing cells demonstrated that they are retained intracellularly. Thus, coexpression of the envelope glycoproteins drastically affects the intracellular transport of the S protein: instead of being transported to the cell surface, S is retained intracellularly by its association with M.


title: Laboratory diagnosis of SARS.

text: The emergence of new viral infections of man requires the development of robust diagnostic tests that can be applied in the differential diagnosis of acute illness, or to determine past exposure, so as to establish the true burden of disease. Since the recognition in April 2003 of the severe acute respiratory syndrome coronavirus (SARS-CoV) as the causative agent of severe acute respiratory syndrome (SARS), enormous efforts have been applied to develop molecular and serological tests for SARS which can assist rapid detection of cases, accurate diagnosis of illness and the application of control measures. International progress in the laboratory diagnosis of SARS-CoV infection during acute illness has led to internationally agreed World Health Organization criteria for the confirmation of SARS. Developments in the dissection of the human immune response to SARS indicate that serological tests on convalescent sera are essential to confirm SARS infection, given the sub-optimal predictive value of molecular detection tests performed during acute SARS illness.


title: European risk assessment guidance for infectious diseases transmitted on aircraft--the RAGIDA project.

text: In order to assist national public health authorities in the European Union to assess the risks associated with the transmission of infectious agents on board aircrafts, the European Centre for Disease Prevention and Control initiated in 2007 the RAGIDA project (Risk Assessment Guidance for Infectious Diseases transmitted on Aircraft). RAGIDA consists of two parts: the production of a systematic review and a series of disease-specific guidance documents. The systematic review covered over 3,700 peer-reviewed articles and grey literature for the following diseases: tuberculosis, influenza, severe acute respiratory syndrome (SARS), invasive meningococcal disease, measles, rubella, diphtheria, Ebola and Marburg haemorrhagic fevers, Lassa fever, smallpox and anthrax. In addition, general guidelines on risk assessment and management from international aviation boards and national and international public health agencies were systematically searched. Experts were interviewed on case-based events by standardised questionnaires. Disease-specific guidance documents on tuberculosis, SARS, meningococcal infections, measles, rubella, Ebola and Marburg haemorrhagic fevers, Lassa fever, smallpox and anthrax were the result of consultations of disease-specific expert panels. Factors that influence the risk assessment of infectious disease transmission on board aircrafts and decision making for contact tracing are outlined.


title: Murine coronavirus requires lipid rafts for virus entry and cell-cell fusion but not for virus release.

text: Thorp and Gallagher first reported that depletion of cholesterol inhibited virus entry and cell-cell fusion of mouse hepatitis virus (MHV), suggesting the importance of lipid rafts in MHV replication (E. B. Thorp and T. M. Gallagher, J. Virol. 78:2682-2692, 2004). However, the MHV receptor is not present in lipid rafts, and anchoring of the MHV receptor to lipid rafts did not enhance MHV infection; thus, the mechanism of lipid rafts involvement is not clear. In this study, we defined the mechanism and extent of lipid raft involvement in MHV replication. We showed that cholesterol depletion by methyl beta-cyclodextrin or filipin did not affect virus binding but reduced virus entry. Furthermore, MHV spike protein bound to nonraftraft membrane at 4 degrees C but shifted to lipid rafts at 37 degrees C, indicating a redistribution of membrane following virus binding. Thus, the lipid raft involvement in MHV entry occurs at a step following virus binding. We also found that the viral spike protein in the plasma membrane of the infected cells was associated with lipid rafts, whereas that in the Golgi membrane, where MHV matures, was not. Moreover, the buoyant density of the virion was not changed when MHV was produced from the cholesterol-depleted cells, suggesting that MHV does not incorporate lipid rafts into the virion. These results indicate that MHV release does not involve lipid rafts. However, MHV spike protein has an inherent ability to associate with lipid rafts. Correspondingly, cell-cell fusion induced by MHV was retarded by cholesterol depletion, consistent with the association of the spike protein with lipid rafts in the plasma membrane. These findings suggest that MHV entry requires specific interactions between the spike protein and lipid rafts, probably during the virus internalization step.


Influenza immunization data: can we make order out of chaos?


title: Cloning and sequence analysis of the spike gene from several feline coronaviruses.

text: The DNA sequence encoding the spike gene from the DF2 strain of Type II feline infectious peritonitis virus (FIPV), a temperature sensitive FIPV virus (TS-DF2) and an isolate of feline enteric coronavirus (FECV 1683) were determined. Comparison of the published WSU 1146 and DF2 FIPV S genes showed that the viruses shared a high degree of homology (99.6%). Likewise, the S gene of the virulent DF2 FIPV virus was closely conserved to that isolated from the vaccine virus strain, TS-DF2 FIPV. In contrast, the FECV S gene had numerous DNA and amino acid differences when compared to the virulent FIPV sequences. Sequence differences among the feline coronavirus isolates were localized to the amino-terminus region of the S gene.


title: Metronidazole; a Potential Novel Addition to the COVID-19 Treatment Regimen.

text: Coronavirus disease 2019 or COVID-19 has rapidly emerged as a global pandemic. This viral infection involves the upper respiratory tract and could lead to severe pneumonia with respiratory distress or even death. Certain studies have found higher initial plasma levels of most pro-inflammatory cytokines during the course of the infection. In this context, both in vitro and in vivo studies have revealed that metronidazole could decrease the levels of several cytokines, which are known to increase during the COVID-19 infection, including interleukin (IL)8, IL6, IL1B, tumor necrosis factor (TNF)α, IL12, IL1α, and interferon (IFN)γ, as well as the levels of C-reactive protein (CRP) and neutrophil count. Furthermore, the drug could decrease neutrophil-generated reactive oxygen species during inflammation. Metronidazole could counteract majority of the immunopathological manifestations of the COVID-19 infection. Therefore, studies with a large sample size are required to determine the efficacy of metronidazole in the treatment of COVID-19 infection.


title: Adoptively transferred acute and chronic relapsing autoimmune encephalomyelitis in the PL/J mouse and observations on altered pathology by intercurrent virus infection.

text: PL/J mice developed chronic relapsing experimental allergic encephalomyelitis (EAE) after receiving syngeneic guinea pig basic protein (GPBP)-immune lymph node cells or spleen cells which were cultured in the presence of GPBP or the encephalitogenic N-terminal peptide of GPBP. The presence of L3T4+ cells and in vitro proliferation in response to GPBP are required for the successful transfer. Pathologically, passively transferred EAE in the virus-free PL/J strain was characterized by an infiltration in the central nervous system by small lymphocytes, followed by the appearance of macrophages, and subsequently by primary demyelination. These findings were similar to those previously observed in chronic relapsing EAE in SJL/J mice. However, in some experiments pathologic examination of the spinal cord showed large demyelinating lesions which were necrotic and infiltrated with eosinophilic polymorphonuclear leukocytes. Sera from mice with this pathology contained antibodies to murine hepatitis virus and extensive search identified a few areas of coronavirus replication. The pathology of autoimmune mediated demyelination may be altered in the presence of coronavirus infection but the clinical pattern of EAE expression did not differ between virus-free and coronavirus-infected mice.


Taiwanese scientists find genetic link to SARS.


title: [B cell subsets and the expression of CD40 in peripheral blood of patients with severe acute respiratory syndrome].

text: OBJECTIVE To explore the changing patterns of B cell subsets and the expression of CD40 on B cells in the course of severe acute respiratory syndrome (SARS) and their relationship to the severity of the disease. METHODS Peripheral blood from 162 cases of SARS, 30 cases of mycoplasma pneumonia and 30 healthy adult volunteers were measured for the number of total B cells (CD19+ B cell), CD5 positive B cells (B1 cell) and mean fluorescent intensity of CD40 on B cell (CD40MF) using tri-color flow cytometry to analyze the distribution of the above parameters in SARS, mycoplasma pneumonia and healthy adult volunteers and their relation to the severity of SARS. RESULTS The number of total B cells of clinically diagnosed SARS was (292 +/- 181) x 10(6)/L, significantly greater than that of healthy adults which was (200 +/- 65) x 10(6)/L (F = 6.17, P < 0.05) but smaller than that of patients infected with mycoplasma pneumonia which was (359 +/- 168) x 10(6)/L (F = 6.28, P < 0.05). During day 11 - 20 of the disease, the number of B1 cells of SARS patients was (24 +/- 14) x 10(6)/L smaller than that of healthy adults which was (39 +/- 20) x 10(6)/L (F = 4.23, P < 0.05). In the early stage of SARS (within 10 days), the expression of CD40 decreased significantly as compared with healthy adults (F = 5.13, P < 0.05). The number of B cells in severe SARS and mild cases were (347 +/- 156) x 10(6)/L and (268 +/- 211) x 10(6)/L respectively. The difference was statistically significant (F = 7.11, P < 0.01). CONCLUSIONS B cells, B1 cells and the expression of CD40 on B cells were involved in the pathogenesis of SARS. The B cell count correlated with the severity of SARS. The monitoring of B cells, B1 cells and the expression of CD40 on B cells is useful in the differential diagnosis of SARS.


title: [Epstein-Barr virus detection in oral hairy leukoplakia in AIDS patients, in leukoplakias and on normal tongue epithelia in HIV-1-negative patients].

text: In 3 patients seropositive for HIV 1 (2 homosexual men and 1 female drug addict), we found "oral hairy leukoplakia" on the lateral margin of their tongues. DNA of Epstein-Barr virus (EBV) and Human Papillomavirus (HPV) were identified in cellular DNA by molecular-virological means using Southern blot hybridization and/or DNA/DNA filter hybridization on epithelial cells taken by swabs from the epithelium of the tongue. The results were compared with those obtained from the identical anatomical sites of 2 HIV-1-seropositive patients without "oral hairy leukoplakia", 3 clinically healthy HIV-1-seronegative patients, and 2 HIV-1-negative patients suffering from oral leukoplakia on their lateral tongues. In all 3 cases investigated, EBV DNA was detected in cells taken from "oral hairy leukoplakia" by filter in situ hybridization. In addition, EBV DNA was found in cells taken from clinically normal epithelia of the tongue as well as in those taken from the 2 leukoplakias mentioned. EBV DNA was regularly detected on the lateral margin of the tongue, but only in one case on the pharyngeal mucosa. Electronmicroscopy on ultrathin sections revealed particles of herpes virus (EBV), particles of HPV, in one case particles of retrovirus, and in another coronavirus and diplococci.


The revised international health regulations and restraint of national health measures.


title: [Clinical features and therapy of 106 cases of severe acute respiratory syndrome].

text: OBJECTIVE To investigate the clinical characteristics of severe acute respiratory syndrome (SARS) and find out its effective treatment. METHODS A total of 106 cases of SARS were analyzed prospectively. RESULTS In this group, 56 were male and 50 female, aged from 15 to 81 years [average (36 +/- 10) years]. Common symptoms included fever (98.1%), chills (75.5%), cough (71.7%), headache and breathless (both 43.4%), diarrhea (24.5%) and rare rales in the lungs (11.2%). Laboratory test showed leukopenia (34.0%), lymphopenia (81.1%) and an extraordinary decrease of CD(4)(+) T cells (98.1%). Other rare abnormalities included liver injury (elevated alanine aminotransferase in 7.6%) and thrombocytopenia (3.8%). Almost all patients suffered from hypoxemia (PaO(2) less than 90 mm Hg in 90.2%, less than 70 mm Hg in 28.6%). Chest radiographs showed that unilateral focal patchy involvement in 34.0% of the patients, and unilateral multifocal or bilateral involvement were 11.3% and 46.2% respectively. Treatment regimens included small doses of steroids (methylprednisolone 40-80 mg, q12 h recommended) accompanied with broad-spectrum antibiotics such as the second generation of cephalosporins and macrolides and some other antiviral drugs. Meanwhile, emphasis was placed on oxygen support and coping with their underlying diseases. CONCLUSIONS SARS has various presentations of clinical features and laboratory tests. Detection of CD(4)(+) T cell count is beneficial to diagnose SARS in early stage. Effective treatment includes various regimens, oxygen support and small doses of steroids.


title: International adoption: infectious diseases issues.

text: Nearly 220,000 children have been adopted from other countries by American parents since 1986. Approximately 65,000 children have arrived from China and Russia, mostly in the past 6 years. Most of these children reside in orphanages before adoption, where they may experience malnutrition, environmental deprivation, neglect, and exposure to infectious diseases. After arrival to the United States, international adoptees should undergo specialized screening evaluation for infectious diseases and other conditions. Infectious conditions of special concern include hepatitis B and C, syphilis, human immunodeficiency virus infection, tuberculosis, and presence of intestinal parasites. Before the adoption occurs, the infectious disease consultant may be asked to assist the primary care provider and the adoptive family with advice about travel and review of preadoptive medical records. After the adoption, the infectious diseases consultant may be asked to assess the adequacy of the child's vaccination record from the birth country and to assist in screening, evaluation, and management of infectious diseases.


title: A new animal model for relapsing polychondritis, induced by cartilage matrix protein (matrilin-1).

text: Relapsing polychondritis (RP) differs from rheumatoid arthritis (RA) in that primarily cartilage outside diarthrodial joints is affected. The disease usually involves trachea, nose, and outer ears. To investigate whether the tissue distribution of RP may be explained by a specific immune response, we immunized rats with cartilage matrix protein (matrilin-1), a protein predominantly expressed in tracheal cartilage. After 2-3 weeks, some rats developed a severe inspiratory stridor. They had swollen noses and/or epistaxis, but showed neither joint nor outer ear affection. The inflammatory lesions involved chronic active erosions of cartilage. Female rats were more susceptible than males. The disease susceptibility was controlled by both MHC genes (f, l, d, and a haplotypes are high responders, and u, n, and c are resistant) and non-MHC genes (the LEW strain is susceptible; the DA strain is resistant). However, all strains mounted a pronounced IgG response to cartilage matrix protein. The initiation and effector phase of the laryngotracheal involvement causing the clinical symptoms were shown to depend on alphabeta T cells. Taken together, these results represent a novel model for RP: matrilin-1-induced RP. Our findings also suggest that different cartilage proteins are involved in pathogenic models of RP and RA.


docid: kvdeznws

title: Production and characterization of egg yolk antibodies against bovine alimentary tract pathogens.

text: Aim of the study was to evaluate the effect of immunization of hens with bovine vaccines (C, R, T) on the course of IgY antibodies production against selected bovine E. coli strains, rota- and coronaviruses in egg yolk in farm conditions. The hens (40 individuals per group) were vaccinated twice, subcutaneously in four week interval and eggs were harvested once a week. Control group consisted of eggs sampled from non-vaccinated hens located in neighbouring cages. The antibody activity was measured by ELISA. All used vaccines induced the rise of IgY antibody in egg yolks. Based on the duration and the highest level of IgY antibody against bovine alimentary tract pathogens C vaccine was further used in next two trials for vaccination of 1000 hens each time. Double immunization seems to be enough in mounting response against examined pathogens for several weeks. Immunization with C vaccine allowed to harvest eggs with satisfactory levels of E. coli, rotavirus and coronavirus IgY antibodies which may be used to evaluate their protective effect by oral administration in calves.


title: Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.

text: The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein. The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses. It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other. The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides. The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins. The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling. Pulse-chase analysis showed that both proteins were modified, although in different ways. While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme. The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides. O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants. In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars. Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.


docid: vywwiprc

title: Conserved translational frameshift in dsDNA bacteriophage tail assembly genes.

text: A programmed translational frameshift similar to frameshifts in retroviral gag-pol genes and bacterial insertion elements was found to be strongly conserved in tail assembly genes of dsDNA phages and to be independent of sequence similarities. In bacteriophage lambda, this frameshift controls production of two proteins with overlapping sequences, gpG and gpGT, that are required for tail assembly. We developed bioinformatic approaches to identify analogous -1 frameshifting sites and experimentally confirmed our predictions for five additional phages. Clear evidence was also found for an unusual but analogous -2 frameshift in phage Mu. Frameshifting sites could be identified for most phages with contractile or noncontractile tails whose length is controlled by a tape measure protein. Phages from a broad spectrum of hosts spanning Eubacteria and Archaea appear to conserve this frameshift as a fundamental component of their tail assembly mechanisms, supporting the idea that their tail genes share a common, distant ancestry.


title: Genetic control of bacterial suicide: regulation of the induction of PBSX in Bacillus subtilis.

text: PBSX is a phage-like bacteriocin (phibacin) of Bacillus subtilis 168. Bacteria carrying the PBSX genome are induced by DNA-damaging agents to lyse and produce PBSX particles. The particles cannot propagate the PBSX genome. The particles produced by this suicidal response kill strains nonlysogenic for PBSX. A 5.2-kb region which controls the induction of PBSX has been sequenced. The genes identified include the previously identified repressor gene xre and a positive control factor gene, pcf. Pcf is similar to known sigma factors and acts at the late promoter PL, which has been located distal to pcf. The first two genes expressed from the late promoter show homology to genes encoding the subunits of phage terminases.


title: Characterization of a feline infectious peritonitis virus isolate.

text: A virus isolated in cell culture from the spleen of a cat with feline infectious peritonitis was identified by physicochemical, morphological and antigenic criteria as a coronavirus. The feline infectious peritonitis virus was compared in vitro with canine coronavirus, a reported enteric pathogen of dogs. The feline isolate was characterized, by chloroform sensitivity and resistance to 5-iododeoxyuridine, respectively, as containing essential lipid and an RNA genome. Other traits of the isolate included resistance to acidic conditions, heat lability, and resistance to trypsin. Electron microscopy showed viral particles with a structure consistent with that of the prototype of the coronavirus group, infectious bronchitis virus. Indirect immunofluorescence with canine coronavirus monospecific antiserum showed the viral isolate to be antigenically related to canine coronavirus. Specific-pathogen-free cats inoculated by various routes with cell-culture-propagated virus had both clinical symptoms and lesions consistent with feline infectious peritonitis.


title: Monoclonal anti-idiotypic antibodies mimicking the immunodominant epitope of influenza virus haemagglutinin elicit biologically significant immune responses.

text: The MAb IIB4 recognizes an immunodominant epitope on influenza virus haemagglutinin (HA) which is shared by different strains of the human subtype H3. This epitope includes amino acids 198, 199 and 201, as determined by selection of IIB4 escape mutants, and is involved in haemagglutination-inhibition (HI) and virus-neutralization (VN). We have developed anti-idiotypic MAbs (Ab2) that mimic the IIB4 epitope. Two Ab2, 78 and 464, completely inhibited binding of MAb IIB4 to the virus. Nucleotide sequences of VL- and VH-encoding regions were determined for IIB4 and both Ab2. VH of IIB4 and 78 belong to the same IgG family (VII) and show high nucleotide identity (89%). Conversely, VH and VL sequences of both internal image-bearing Ab2 revealed lower degrees of identity (61 and 50%, respectively). Ab2 were used for syngeneic immunization to elicit polyclonal Ab3 responses. Like Ab1, Ab3 immunoprecipitated viral HA and displayed HI and VN activity. The different VN activity of anti-78 and anti-464 in vitro correlated with the affinities of their corresponding Ab2 to IIB4. In vivo immunization with either Ab2 protected 15-37% of mice against lethal influenza infection or delayed dying. In contrast to VN activity of Ab3 in vitro, there was no significant difference between the protection of mice induced by Ab2 78 and 464. We demonstrate, for the influenza model, that active immunization with a single influenza virus HA epitope in the form of its internal image leads to partial protection in vivo.


title: Serological relationships of the subcomponents of human coronavirus strain 229E and mouse hepatitis virus strain 3.

text: Antibodies were raised in rabbits against the structural components of human coronavirus strain 229E and mouse hepatitis virus strain 3, prepared from disrupted virus particles. Hyperimmune sera to the subcomponents showed cross-reactions by enzyme-linked immunosorbent assay between ribonucleoprotein antigens of these viruses, indicating the presence of a common antigen(s). None of the other virus structural components showed any cross-reactivity.


title: Effect of cellular changes and onset of humoral immunity on the replication of porcine reproductive and respiratory syndrome virus in the lungs of pigs.

text: Twenty-two 4- to 5-week-old gnotobiotic pigs were intranasally inoculated with 10(6.0) TCID(50) of porcine reproductive and respiratory syndrome virus (PRRSV) (Lelystad) and euthanized at different time intervals post-inoculation (p.i.). Bronchoalveolar lavage (BAL) cell populations were characterized, together with the pattern of virus replication and appearance of antibodies in the lungs. Total BAL cell numbers increased from 140x10(6) at 5 days p.i. to 948x10(6) at 25 days p.i. and remained at high levels until the end of the experiment. The number of monocytes/macrophages, as identified by monoclonal antibodies 74-22-15 and 41D3, increased two- to fivefold between 9 and 52 days p.i. with a maximum at 25 days p.i. Flow cytometry showed that the population of differentiated macrophages was reduced between 9 and 20 days p.i. and that between the same time interval, both 74-22-15-positive and 41D3-negative cells, presumably monocytes, and 74-22-15- and 41D3-double negative cells, presumably non-phagocytes, entered the alveolar spaces. Virus replication was highest at 7 to 9 days p.i., decreased slowly thereafter and was detected until 40 days p.i. Anti-PRRSV antibodies were detected starting at 9 days p.i. but neutralizing antibodies were only demonstrated in one pig euthanized at 35 days and another at 52 days p.i. The decrease of virus replication in the lungs from 9 days p.i. can be attributed to (i) shortage of susceptible differentiated macrophages, (ii) lack of susceptibility of the newly infiltrated monocytes and (iii) appearance of anti-PRRSV antibodies in the lungs. Neutralizing antibodies may contribute to the clearance of PRRSV from the lungs.


Medical education and COVID-19: a personal view.


Strategies for a successful coronavirus (TGE) vaccine for swine.


docid: xetqooic

title: The 17th International Conference on Antiviral Research.

text: The focus of the 17th International Conference on Antiviral Research was the discovery and development of antiviral agents (chemistry, biology, animal models and clinical trial results) against a variety of human infectious agents including HIV, herpes viruses, hepatitis viruses, respiratory viruses and emerging/re-emerging pathogens. The meeting included the symposium 'Clinical Update on Antiviral Drugs', plenary sessions dedicated to each of the individual classes of infectious agents, a symposium on new developments surrounding emerging pathogens, and three special award lectures, which discussed the history of nucleotide antiviral agents, mechanisms of viral persistence and drug resistance, and the therapy of herpes virus infections. Within each infectious agent session the presentations included those describing the development of new and novel anti-infectives, including research based on the preclinical development of new molecules, and the results of animal modelling and clinical studies on advanced-stage antiviral agents. A summary of the meeting highlights, segregated by infectious agent, will be presented in this review.


title: A subset of porcine reproductive and respiratory syndrome virus GP3 glycoprotein is released into the culture medium of cells as a non-virion-associated and membrane-free (soluble) form.

text: The GP3 protein of the IAF-Klop strain of porcine reproductive and respiratory syndrome virus (PRRSV) was expressed in 293 cells by a recombinant human type 5 adenovirus carrying the open reading frame 3 gene. The protein exhibited a molecular mass of 42 kDa and comigrated with GP3 expressed in PRRSV-infected MARC-145 cells. Removal of N-linked glycans from GP3 resulted in a 27-kDa protein (P3), confirming its highly glycosylated nature. Pulse-chase experiments carried out either in the context of PRRSV infection or upon individual expression of GP3 in 293 cells showed that the protein remains completely endo-beta-N-acetylglucosaminidase H-sensitive even after 4 h of synthesis. Thus, the transport of GP3 was restricted to the premedial Golgi compartment, presumably the endoplasmic reticulum (ER). However, a minor fraction of GP3 was found to be secreted in the culture medium as a soluble membrane-free form. This released protein (sGP3) was readily identified upon individual expression of GP3 in 293 cells as well as in the context of PRRSV infection, albeit at lower levels. The sGP3 migrated as a smear and displayed a molecular mass ranging from 43 to 53 kDa. The unglycosylated form of sGP3 comigrated with its intracellular deglycosylated counterpart, suggesting that the release from the cell of a subset of GP3 did not result from cleavage of a putative membrane-anchor sequence. Strikingly, unlike GP3, the sGP3 acquired Golgi-specific modifications of its carbohydrate side chains and folded into a disulfide-linked homodimer. Brefeldin A treatment completely abolished the release of sGP3, suggesting that the ER-to-Golgi compartment is an obligatory step in cellular secretion of sGP3. In contrast, 10 mM monensin did not prevent sGP3 release but inhibited the terminal glycosylation that confers on the protein its diffuse pattern. Since GP3 was found to be nonstructural in the case of the North American strain, secretion of a minor fraction of GP3 might be an explanation for its high degree of immunogenicity in infected pigs. Furthermore, this secreted protein might be relevant as a model for further studies on the cellular subcompartments involved in the sorting of proteins to the extracellular milieu.


title: Emergency Planning in Ontario's Acute Care Hospitals: A Survey of Board Chairs.

text: BACKGROUND Effective hospital governance depends on proactive board leadership to minimize risk. STUDY AIM To survey hospital board chairs about governance practices, particularly with respect to approval processes for oversight of management preparedness for unforeseen emergencies. METHODS A 2004 survey of hospital managers initially suggested greater board leadership in risk management as a desired strategic priority for Ontario's acute care hospitals. Our literature review and panel process defined 34 best practices in board governance, including two practices explicitly addressing the board's role in preparing for risk. RESULTS Our findings revealed that some boards may not be actively engaged in ensuring that adequate processes are in place to protect against risk. More than one-quarter (n=28, 26.9%) of board chairs reported that they had not approved a management plan to address emergencies. Thirty respondents (28.8%) said they had not approved a process to identify, manage and minimize risks to the hospital's sustainability. Forty-seven respondents (45.2%) said they had not approved both of these two processes. A significant association emerged between boards that had approved both risk preparation strategies and boards that had implemented six key governance practices relating to accountability for leadership and stakeholder communication.


title: Unusual infections and novel therapy in the immunocompromised host.

text: Many of the reports of infection in immunocompromised patients over the past year have been caused by unusual bacterial pathogens or fungi, which are often multiresistant and difficult to treat. Although many difficulties still remain some progress has been made over the past year in the development of new antimicrobial agents and alternative therapies.


Virtual dermatology: a COVID-19 update.


title: Isolation of small viruses resembling astroviruses and caliciviruses from acute enteritis of calves.

text: Small round viruses (SRV) were isolated from the faeces of diarrhoeic calves from three farms. All three SRV preparations caused diarrhoea experimentally in gnotobiotic calves. Each preparation contained viral particles of two morphological types, "astrovirus-like" and "calicivirus-like", and from one preparation the two particle types were separated from each other. The calicivirus-like agent ("Newbury agent") was 33 nm in diameter, and caused diarrhoea in gnotobiotic calves with villous atrophy and D-xylose malabsorption. This virus did not infect cell cultures. The astrovirus-like agent did not cause diarrhoea in two gnotobiotic calves; however, it infected cell cultures (primary calf kidney) and the infected cells immunofluoresced with convalescent gnotobiotic-calf antiserum. The astrovirus-like agents in the three preparations were antigenically related. Experiments in calves showed that there was a degree of cross-protection between the three SRV preparations, as judged by the presence or absence of diarrhoea, but that at least three unrelated pathogens were present.


title: High-dose pulse versus nonpulse corticosteroid regimens in severe acute respiratory syndrome.

text: The treatment of atypical pneumonia, subsequently termed severe acute respiratory syndrome (SARS), is controversial, and the efficacy of corticosteroid therapy is unknown. We have evaluated the clinical and radiographic outcomes of 72 patients with probable SARS (median age 37 years, 30 M), who received ribavirin and different steroid regimens in two regional hospitals. Chest radiographs were scored according to the percentage of lung field involved. Seventeen patients initially received pulse steroid (PS) (methylprednisolone > or =500 mg/day) and 55 patients initially received nonpulse steroid (NPS) (methylprednisolone <500 mg/day) therapy. The cumulative steroid dosage; intensive care unit admission, mechanical ventilation, and mortality rates; and hematologic and biochemical parameters were similar in both groups after 21 days. However, patients in the PS group had less oxygen requirement, better radiographic outcome, and less likelihood of requiring rescue PS therapy than their counterparts. There was no significant difference between the two groups in hemolytic anemia, severe secondary infections, or hematemesis, but patients in the PS group had less hyperglycaemia. Initial use of pulse methylprednisolone therapy appears to be a more efficacious and an equally safe steroid regimen when compared with regimens with lower dosage and should be considered as the preferred steroid regimen in the treatment of SARS, pending data from future randomized controlled trials.


title: Diarrhoeal diseases in the White Mountain Apaches: epidemiologic studies.

text: Acute diarrhoeal disease in children is known to be a major public health problem among native Americans living in reservations in the southwestern part of the United States. This study was undertaken to describe the epidemiology and causative agents of diarrhoea more completely, with the expectation that this information may help in the ultimate control of the disease in this population. Three interrelated epidemiologic studies were carried out in the White Mountain Apache Tribe, Whiteriver, Arizona, during 1981-1985: a three-year longitudinal study on a cohort of 112 newborns, a longitudinal two-year study in a cohort of 200 families, and a case-control study on 1,072 children with diarrhoea attending a medical facility. Both epidemiologic and microbiological patterns of diarrhoeal disease were found to be very similar to those seen in developing countries, indicating the need for basic improvements in sanitation and hygiene in this population.


docid: jgzydupt

title: Comparison of bupivacaine and parecoxib for postoperative pain relief after laparoscopic cholecystectomy: a randomized controlled trial.

text: BACKGROUND Pain is the most common complaint of patients on the first day after laparoscopic cholecystectomy (LC). The aim of this study was to compare the efficacy of local anesthesia with bupivacaine and intravenous parecoxib on postoperative abdominal pain relief up to 24 h after surgery. METHODS One hundred and eighty patients who underwent LC were randomized to one of three groups with sixty patients each: Group A received 50 mg 0.5% bupivacaine subcutaneously at trocar sites before incision closure; Group B received intravenous parecoxib (40 mg) after entering the recovery room; Group C did not receive postoperative analgesia unless needed and was served as control. The postoperative pain at 1, 2, 4, 8, 12, and 24 h after the operation was assessed using a visual analog scale (VAS). Secondary outcomes, including intraoperative and postoperative complications, the incidence of shoulder pain, pethidine requirements, postoperative nausea and vomiting, and hospital stay were also recorded. RESULTS At 1, 2, and 4 hours after surgery, VAS pain scores were significantly lower in group A and B compared with group C (P < 0.05 for all). There was no significant difference among the three groups at 8, 12, and 24 hours after the procedure (P > 0.05 for all). A repeated-measures ANOVA analysis revealed that VAS pain scores over the first 24 hours after LC were significantly lower in group A and B compared with group C (P = 0.014 and P = 0.029 for between-group comparison, respectively). Furthermore, the percentage of patients requiring postoperative rescue analgesics was significantly higher in group C as compared with group A and group B (P = 0.018). CONCLUSION Local anesthesia with bupivacaine and intravenous parecoxib are both effective at decreasing postoperative pain and pethidine requirements after LC.


title: Four proteins processed from the replicase gene polyprotein of mouse hepatitis virus colocalize in the cell periphery and adjacent to sites of virion assembly.

text: The replicase gene (gene 1) of the coronavirus mouse hepatitis virus (MHV) encodes two co-amino-terminal polyproteins presumed to incorporate all the virus-encoded proteins necessary for viral RNA synthesis. The polyproteins are cotranslationally processed by viral proteinases into at least 15 mature proteins, including four predicted cleavage products of less than 25 kDa that together would comprise the final 59 kDa of protein translated from open reading frame 1a. Monospecific antibodies directed against the four distinct domains detected proteins of 10, 12, and 15 kDa (p1a-10, p1a-12, and p1a-15) in MHV-A59-infected DBT cells, in addition to a previously identified 22-kDa protein (p1a-22). When infected cells were probed by immunofluorescence laser confocal microscopy, p1a-10, -22, -12, and -15 were detected in discrete foci that were prominent in the perinuclear region but were widely distributed throughout the cytoplasm as well. Dual-labeling experiments demonstrated colocalization of the majority of p1a-22 in replication complexes with the helicase, nucleocapsid, and 3C-like proteinase, as well as with p1a-10, -12, and -15. p1a-22 was also detected in separate foci adjacent to the replication complexes. The majority of complexes containing the gene 1 proteins were distinct from sites of accumulation of the M assembly protein. However, in perinuclear regions the gene 1 proteins and nucleocapsid were intercalated with sites of M protein localization. These results demonstrate that the complexes known to be involved in RNA synthesis contain multiple gene 1 proteins and are closely associated with structural proteins at presumed sites of virion assembly.


title: [Analysis of acute physiology and chronic health evaluation III in the 54 patients with severe acute respiratory syndrome].

text: OBJECTIVE To evaluate the acute physiology and chronic health evaluation III (APACHE III) in the severe acute respiratory syndrome (SARS) patients and its significance in prognosis. METHODS The clinical data of 54 SARS patients, including survivors (43 cases) and nonsurvivors (11 cases) were collected and evaluated with APACHE III scoring system. The correlation of scores and prognosis was evaluated. RESULTS The scores of the nonsurvivors were higher remarkably than those of the surviving group(P<00.01). The scores of the acute physiology score (APS) and year score (YS) in the death group were higher obviously than those of the surviving group(both P<0.01). Elderly patients with severe disease had a high mortality. The scores of APACHE III had positive correlation with the over all fatality rate. When the scores of APACHE III was higher than 60, the fatality rate increased obviously. CONCLUSION The scores of APACHE III in the SARS are correlated with the patient's condition and prognosis.


title: Pneumonia in the elderly.

text: PURPOSE OF REVIEW This review provides an update on new data about the etiology, clinical characteristics, outcomes, and prevention of community-acquired pneumonia. The impact of severe acute respiratory syndrome in older persons is also addressed. RECENT FINDINGS Streptococcus pneumoniae remains the most important cause of community-acquired pneumonia in the very elderly (80 years and over). Pneumococcus is also an important etiologic agent of pneumonia in residents of long-term care facilities. Clinical signs and symptoms of community-acquired pneumonia are less distinct in this group compared with younger patients. New data about influenza immunization reveal better mucosal immunity with the intranasal vaccine compared with the intramuscular vaccine. A review of clinical trial evidence differs from observational studies that demonstrate a clear benefit of the polysaccharide pneumococcal vaccine in the elderly. The prospect of severe acute respiratory syndrome in older adults is the most important new issue to emerge, as older adults are at increased risk from complications and death. SUMMARY There are many challenges in preventing and managing community-acquired pneumonia in the elderly. S. pneumoniae remains the most important cause. The clinical presentation of pneumonia in the very elderly can be nonspecific. Given the safety profile of the vaccine and supportive observational evidence, public health policymakers should continue to support use of the polysaccharide pneumococcal vaccine. However, research into new vaccine strategies, including use of conjugate or common antigen vaccines, is a priority.


Viral aetiology of acute respiratory illnesses in patients with a suspicion of severe acute respiratory syndrome (SARS) in Switzerland.


title: Viruses and bacteria in the etiology of the common cold.

text: Two hundred young adults with common colds were studied during a 10-month period. Virus culture, antigen detection, PCR, and serology with paired samples were used to identify the infection. Viral etiology was established for 138 of the 200 patients (69%). Rhinoviruses were detected in 105 patients, coronavirus OC43 or 229E infection was detected in 17, influenza A or B virus was detected in 12, and single infections with parainfluenza virus, respiratory syncytial virus, adenovirus, and enterovirus were found in 14 patients. Evidence for bacterial infection was found in seven patients. Four patients had a rise in antibodies against Chlamydia pneumoniae, one had a rise in antibodies against Haemophilus influenzae, one had a rise in antibodies against Streptococcus pneumoniae, and one had immunoglobulin M antibodies against Mycoplasma pneumoniae. The results show that although approximately 50% of episodes of the common cold were caused by rhinoviruses, the etiology can vary depending on the epidemiological situation with regard to circulating viruses. Bacterial infections were rare, supporting the concept that the common cold is almost exclusively a viral disease.


Misdiagnostic analysis of clinically diagnosed severe acute respiratory syndrome after following up 197 convalescent patients.


title: Structure of a proteolytically resistant core from the severe acute respiratory syndrome coronavirus S2 fusion protein.

text: A coronavirus (CoV) has recently been identified as the causative agent of the severe acute respiratory syndrome (SARS) in humans. CoVs enter target cells through fusion of viral and cellular membranes mediated by the viral envelope glycoprotein S. We have determined by x-ray crystallography the structure of a proteolytically stable core fragment from the heptad repeat (HR) regions HR1 and HR2 of the SARS-CoV S protein. We have also determined the structure of an HR1-HR2 S core fragment, containing a shorter HR1 peptide and a C-terminally longer HR2 peptide that extends up to the transmembrane region. In these structures, three HR1 helices form a parallel coiled-coil trimer, whereas three HR2 peptides pack in an oblique and antiparallel fashion into the coiled-coil hydrophobic grooves, adopting mixed extended and alpha-helical conformations as in postfusion paramyxoviruses F proteins structures. Our structure positions a previously proposed internal fusion peptide adjacent to the N-terminus of HR1. Peptides from the HR2 region of SARS-CoV S have been shown to inhibit viral entry and infection in vitro. The structures presented here can thus open the path to the design of small-molecule inhibitors of viral entry and candidate vaccine antigens against this virus.


title: Comparative analysis of innate immune responses following infection of newborn calves with bovine rotavirus and bovine coronavirus.

text: Bovine rotavirus (BRV) and bovine coronavirus (BCV) are important causes of diarrhoea and death in newborn calves. Although these viruses belong to distinct viral classes, they both infect intestinal epithelial cells and induce similar clinical symptoms. Rotavirus usually causes an acute infection, but coronavirus infection can persist and reoccur in adults. Differences in viral structure and clinical outcome prompted us to postulate that innate mucosal immune responses would be markedly different following rotavirus and coronavirus infections. To address this hypothesis, gene expression following BRV and BCV infection was analysed in surgically prepared intestinal loops from 1-day-old colostrum-deprived calves. Gene expression was profiled at 18 h post-infection using bovine cDNA microarrays; the majority of differentially expressed significant genes were associated with the cell cycle and innate immune responses. A select group of these genes was validated by quantitative real-time PCR (qRT-PCR). The expression of genes associated with interferons (IFNs), cytokines and Toll-like receptors, which were not present on the microarray, was analysed further by qRT-PCR. Strong activation of TLR3, IL-6 and p65 was observed in BRV-infected host tissues, but not in tissues infected with BCV. Both viruses also downregulated IFN- and pro-inflammatory cytokine-associated pathways. In vitro studies confirmed that IFN inhibited viral replication. All of these results together suggested either that very early events of host responses at 18 h post-infection were being observed, or that both viruses have unique effective strategies to evade host immune responses.


docid: vftisquo

title: Information control in time of crisis: the framing of SARS in China-based newspapers and Internet sources.

text: A content analysis examined 727 SARS news stories from a commercial China-based Web portal, a national Chinese newspaper, and a Chinese regional newspaper. Coding included news frames (health severity, human interest, economic, attribution of responsibility, denial) and the type of cited sources (domestic and international). MANOVA analyses indicated that the national newspaper used the health severity frame more often than the regional newspaper. The China-based Web portal used the economic frame and international sources significantly more often than either newspaper. Results are discussed in terms of the future of the Internet in China as a tool to mitigate the spread of infectious diseases through rapid information dissemination.


title: A cDNA clone from a defective RNA of citrus tristeza virus is infective in the presence of the helper virus.

text: A naturally occurring defective RNA of 2379 nt (D2.3) from the VT strain of citrus tristeza closterovirus (CTV) was cloned and sequenced. The D2.3 RNA is a fusion of two regions of 1521 and 858 nt from the 5' and 3' ends of the CTV genome, respectively. A cDNA clone of D2.3 RNA was tagged by the insertion of a 0.47 kb chimeric DNA fragment and the recombinant cDNA was inserted downstream of the cauliflower mosaic virus 35S promoter. The resulting construct was bombarded into CTV-infected tissue, which was then grafted onto virus-free plants. The presence of recombinant RNA in systemically infected leaves was demonstrated by RT-PCR. Sequencing the RT-PCR products synthesized from double-stranded RNA confirmed the presence of the chimeric segment used for tagging. This is the first report of an infectious cDNA molecule derived from CTV D-RNA.


title: Risks of transfusion-transmitted infections: 2003.

text: PURPOSE OF REVIEW While the risks of transfusion-transmitted human immunodeficiency virus, hepatitis C virus, and human T-cell leukemia virus I/II continue to decrease, additional threats to transfusion safety are posed by emerging "new" infectious diseases. RECENT FINDINGS Following the introduction of nucleic acid testing for human immunodeficiency virus and hepatitis C virus, the American Red Cross estimates the risk of transfusion-transmitted human immunodeficiency virus to be 1:1,215,000 (per unit transfused) and 1:1,935,000 for transfusion-transmitted hepatitis C virus. Hepatitis B virus nucleic acid testing has not been implemented, and the risk of transfusion-transmitted hepatitis B virus in the United States remains relatively high at an estimated 1:205,000. The risk of transfusion-transmitted human T-cell leukemia virus I/II is 1:2,993,000, based on Red Cross estimates. Nucleic acid testing for West Nile virus began in the United States in 2003 under an investigational new drug program. No approved laboratory tests are available to screen the blood for Chagas disease, malaria, severe acute respiratory syndrome, or variant Creutzfeldt-Jakob disease. SUMMARY Prevention of these potential transfusion-transmitted infections is addressed by deferring potential donors whose personal behaviors or travel histories place them at risk.


COVID-19: Has the Standard of Care Changed and Are Providers Immune from Liability?


title: Tracing Airline Travelers for a Public Health Investigation: Middle East Respiratory Syndrome Coronavirus (MERS-CoV) Infection in the United States, 2014.

text: OBJECTIVE CDC routinely conducts contact investigations involving travelers on commercial conveyances, such as aircrafts, cargo vessels, and cruise ships. METHODS The agency used established systems of communication and partnerships with other federal agencies to quickly provide accurate traveler contact information to states and jurisdictions to alert contacts of potential exposure to two travelers with Middle East Respiratory Syndrome Coronavirus (MERS-CoV) who had entered the United States on commercial flights in April and May 2014. RESULTS Applying the same process used to trace and notify travelers during routine investigations, such as those for tuberculosis or measles, CDC was able to notify most travelers of their potential exposure to MERS-CoV during the first few days of each investigation. CONCLUSION To prevent the introduction and spread of newly emerging infectious diseases, travelers need to be located and contacted quickly.


title: [Cryptosporidium: a "new" factor in the aetiology of neonatal diarrhoea in calves].

text: In recent years, Cryptosporidium, a coccidian parasite of the intestine, has been associated with neonatal diarrhoea in calves. Current knowledge of the organism with regard to classification, host-specificity, life cycle, pathology, incidence, clinical treatment, prevention and diagnosis is reviewed in the present paper. In addition, Cryptosporidium infection in calves in the Netherlands is reported for the first time. The typical 4 micrometer oocysts were observed on microscopic examination of te faeces of a three-week-old calf with severe diarrhoea and in the faeces of three other clinical normal calves of the same group. Tests performed simultaneously for the presence of rotaviruses, bovine coronaviruses or K99-positive E. coli were negative. Histological and electron-microscopic studies of the intestines of orally inoculated baby mice served to verify the diagnosis of cryptosporidiosis.


Addition of a single glycosylation site to hAPN blocks human coronavirus-229E receptor activity.


title: [COVID-19 associated severe psychotic relapse].

text: This is a case report of a 42-year-old patient, who was admitted to hospital with clinical pneumonia. The only positive test result was for SARS-CoV-2. After a few days of treatment, the patient was discharged in stable condition for isolation at home. The patient had previously had contact with the mental health services and was diagnosed with catatonic schizophrenia but had been without outpatient contact or treatment for the last four years. After seven days he was admitted in a state of severe psychosis. This begs the question if the psychotic relapse was triggered by a neuroinflammatory response caused by SARS-CoV-2.


title: Transmissible gastroenteritis virus infection: a vanishing specter.

text: About twenty years ago, a new coronavirus, porcine respiratory coronavirus (PRCoV), was detected in swine herds. This virus is related to transmissible gastroenteritis virus (TGEV); however, it is not enteropathogenic but causes only minor respiratory symptoms. As PRCoV shares some epitopes for neutralizing antibodies with TGEV, it acts like a nature-made vaccine against TGEV resulting in a drastic reduction of TGE outbreaks in Europe. A major difference between the two porcine coronaviruses is a large deletion in the surface protein S gene of PRCoV. Because of this structural difference, TGEV but not PRCoV has a sialic acid binding activity that allows the attachment to mucins and mucin-type glycoproteins. The sialic acid binding activity may allow TGEV to overcome the mucus barrier in the gut and to get access to the intestinal epithelium for initiation of infection.


title: Surgical vs conventional therapy for weight loss treatment of obstructive sleep apnea: a randomized controlled trial.

text: CONTEXT Obstructive sleep apnea (OSA) is strongly related to obesity. Weight loss is recommended as part of the overall management plan for obese patients diagnosed with OSA. OBJECTIVE To determine whether surgically induced weight loss is more effective than conventional weight loss therapy in the management of OSA. DESIGN, SETTING, AND PATIENTS A randomized controlled trial of 60 obese patients (body mass index: >35 and <55) with recently diagnosed (<6 months) OSA and an apnea-hypopnea index (AHI) of 20 events/hour or more. These patients had been prescribed continuous positive airway pressure (CPAP) therapy to manage OSA and were identified via accredited community sleep clinics. The trial was conducted between September 2006 and March 2009 by university- and teaching hospital-based clinical researchers in Melbourne, Australia. Patients with obesity hypoventilation syndrome, previous bariatric surgery, contraindications to bariatric surgery, or significant cardiopulmonary, neurological, vascular, gastrointestinal, or neoplastic disease were excluded. INTERVENTIONS Patients were randomized to a conventional weight loss program that included regular consultations with a dietitian and physician, and the use of very low-calorie diets as necessary (n = 30) or to bariatric surgery (laparoscopic adjustable gastric banding; n = 30). MAIN OUTCOME MEASURES The primary outcome was baseline to 2-year change in AHI on diagnostic polysomnography scored by staff blinded to randomization. Secondary outcomes were changes in weight, CPAP adherence, and functional status. RESULTS Patients lost a mean of 5.1 kg (95% CI, 0.8 to 9.3 kg) in the conventional weight loss program compared with 27.8 kg (95% CI, 20.9 to 34.7 kg) in the bariatric surgery group (P < .001). The AHI decreased by 14.0 events/hour (95% CI, 3.3 to 24.6 events/hour) in the conventional weight loss group and by 25.5 events/hour (95% CI, 14.2 to 36.7 events/hour) in the bariatric surgery group. The between-group difference was -11.5 events/hour (95% CI, -28.3 to 5.3 events/hour; P = .18). CPAP adherence did not differ between the groups. The bariatric surgery group had greater improvement in the Short Form 36 physical component summary score (mean, 9.3 [95% CI, 0.5 to 18.0]; P = .04). CONCLUSION Among a group of obese patients with OSA, the use of bariatric surgery compared with conventional weight loss therapy did not result in a statistically greater reduction in AHI despite major differences in weight loss. TRIAL REGISTRATION anzctr.org Identifier: 12605000161628.


Advances in infectious diseases.


title: [Chest X-ray changes after discontinuation of glucocorticosteroids treatment on severe acute respiratory syndrome (5 cases report)].

text: OBJECTIVE To investigate the effect of glucocorticosteroids on severe acute respiratory syndrome (SARS) and chest X ray changes after discontinuation of glucocorticosteroids treatment. METHODS To retrospectively study the clinical characteristics of 5 cases with SARS and chest X ray changes after discontinuation of glucocorticosteroids. RESULTS 5 cases were medical workers and had exposure to SARS patients. The incubation periods were 2 to 7 days. They had high fever from the onset of the disease. There were lung infiltrates 1 to 6 days after fever and then methylprednisolone was given to them. Two or three days after glucocorticosteroids treatment, clinical symptoms in these patients were disappeared. The clinical symptoms in case 1, cases 2, case 4 and case 5 weren't deteriorated after discontinuation of methylprednisolone while the presentation in chest X ray was progressed 3-6 days later. No more methylprednisolone was given to the patients and the infiltrates in chest X ray was gradually absorbed in 5-11 days. The temperature in case 3 was elevated again and the shadow in chest X ray was progressed after discontinuation of methylprednisolone due to the low dosage of glucocorticosteroids. When the dosage of methylprednisolone was increased, the temperature tended to be normal and the lung infiltrates began to absorb gradually. CONCLUSION For the patients with non-severe SARS, if the symptoms were disappeared after discontinuation of glucocorticosteroids, the lung shadow may be progressed. we can closely observe the condition of the disease and no more glucocortisteroids needed to be given to the patients. The lung shadow could be absorbed gradually.


title: Specific plant terpenoids and lignoids possess potent antiviral activities against severe acute respiratory syndrome coronavirus.

text: In this study, 221 phytocompounds were evaluated for activity against anti-severe acute respiratory syndrome associated coronavirus (SARS-CoV) activities using a cell-based assay measuring SARS-CoV-induced cytopathogenic effect on Vero E6 cells. Ten diterpenoids (1-10), two sesquiterpenoids (11 and 12), two triterpenoids (13 and 14), five lignoids (15-19), curcumin (20), and reference controls niclosamide (21) and valinomycin (22) were potent inhibitors at concentrations between 3.3 and 10 microM. The concentrations of the 22 compounds to inhibit 50% of Vero E6 cell proliferation (CC50) and viral replication (EC50) were measured. The selective index values (SI = CC50/EC50) of the most potent compounds 1, 5, 6, 8, 14, and 16 were 58, >510, 111, 193, 180, and >667, respectively. Betulinic acid (13) and savinin (16) were competitive inhibitors of SARS-CoV 3CL protease with Ki values = 8.2 +/- 0.7 and 9.1 +/- 2.4 microM, respectively. Our findings suggest that specific abietane-type diterpenoids and lignoids exhibit strong anti-SARS-CoV effects.


title: Onstep repair of inguinal hernias.

text: Inguinal hernias are a protrusion of the peritoneum through a weakening in the groin in which abdominal content (intestines or fat) can herniate and cause a bulge. Inguinal hernias can be painful and require surgery. Worldwide, approximately 20 million patients are operated each year, with 10,000 in Denmark. The repair of inguinal hernias causes pain and 16% of patients experience chronic pain six months after the standard, open, mesh-based Lichtenstein technique. Therefore, surgeons are trying to improve the techniques by finding new ways of operating. The Onstep method was a new method for the repair of inguinal hernias, presented along with excellent results regarding pain, recurrence and complications. However, the technique had not been tested outside the department of the inventors. The overall aim was to clarify whether the Onstep technique should be implemented on a larger scale outside the departments of the inventors. Six papers are included in this thesis: a systematic review, a protocol article, three reports on the Onstep versus Lichtenstein trial, and finally a focus group interview. The systematic review identified nine different methods of placing a preperitoneal mesh through and open anterior approach. In general, the techniques seem to provide good results regarding pain and discomfort, but more studies are needed. The protocol article describes the randomized, double blinded Onstep versus Lichtenstein study, with focus on the statistical analysis and sample size calculations. Four separate sample size calculations were conducted, making several primary outcomes possible. The three reports of the Onstep versus Lichtenstein study reported on early postoperative outcomes, on chronic pain, and lastly on sexual dysfunction. The overall findings from the trial demonstrated that there were no differences between the Onstep and the Lichtenstein technique regarding early and chronic pain (30 days, six months, and 12 months). However, for the group of patients operated with the Onstep technique, fewer patients experienced pain during sexual activity. The focus group interview was done with experienced surgeons teaching the Onstep technique. They described their experience, thoughts, and concerns regarding teaching the technique. The results from the focus group interview can be used to guide future trainings sessions. In this thesis the Onstep technique has been investigated in comparison with the Lichtenstein technique, but the results have not been as promising as the initial studies from the inventors. However, implementation of the Onstep technique outside the departments of the inventors is unlikely to result in increased risk of complications. Furthermore, the Onstep technique could possibly benefit patients by reducing the risk of pain during sexual activity.


title: Long-term outcome of acute respiratory distress syndrome caused by severe acute respiratory syndrome (SARS): an observational study.

text: OBJECTIVE We examined long-term outcome of pulmonary function, exercise capacity and health-related quality of life (HRQoL) in patients with acute respiratory distress syndrome (ARDS) caused by severe acute respiratory syndrome (SARS). METHODS 59 critically ill patients with ARDS caused by SARS between March and July 2003 were studied prospectively and followed up for 1 year. Thirty-six underwent pulmonary function testing and a 6-minute walk test, and 35 underwent HRQoL evaluation by Short Form-36 questionnaire at 3, 6, and 12 months after illness onset. RESULTS Mean age was 47 (SD, 15.7) years. Median APACHE II score was 10 (interquartile range [IQR], 7-12). Only 47% required invasive mechanical ventilation. Median admission and worst PaO2/FIO2 ratio were 142 (IQR, 94-177) mmHg and 86 (IQR, 66-122) mmHg, respectively. Median stay in ICU and hospital were 9 (IQR, 5-20) and 31 (IQR, 20-54) days, respectively. Mortality was 24% at hospital discharge and at 1 year. Mean lung volumes and spirometric measurements were nearly normal by 6 months. Except for diffusion capacity adjusted for haemoglobin concentration (DLCO) measured at 12 months, there was no significant difference in pulmonary function measurement between those who had mechanical ventilation and those who did not. The 6-minute walk distance (6MWD) improved from 3 to 6 months, with no further significant change. Younger patients had near normal HRQoL by 6 months. Those aged over 40 years had impaired HRQoL at multiple domains even at 12 months. At 12 months, forced expiratory volume in 1 second, forced vital capacity and 6MWD correlated significantly with multiple SF-36 domain scores. DLCO correlated significantly only with domain scores reflecting physical function. CONCLUSIONS The mortality of SARS-related ARDS is similar to the mortality of ARDS from other causes. A substantial number of patients with SARS-related ARDS survived without receiving mechanical ventilation. Patients had good recovery of pulmonary function and HRQoL.


title: Carboxypeptidase D is an avian hepatitis B virus receptor.

text: The receptor molecules for human and animal hepatitis B viruses have not been defined. Previous studies have described a 170 to 180 kDa molecule (p170 or gp180) that binds in vitro to the pre-S domain of the large envelope protein of duck hepatitis B virus (DHBV); cDNA cloning revealed the binding protein to be duck carboxypeptidase D (DCPD). In the present study, the DCPD cDNA was transfected into several nonpermissive human-, monkey-, and avian species-derived cell lines. Cells transfected with a plasmid encoding the full-length DCPD protein bound DHBV particles, whereas cells expressing truncated versions of DCPD protein that fail to bind the pre-S protein did not. The DHBV binding to DCPD-reconstituted cells was blocked by a monoclonal antibody that neutralizes DHBV infection of primary duck hepatocytes (PDH) and also by a pre-S peptide previously shown to inhibit DHBV infection of PDH. In addition to promoting virus binding, DCPD expression was associated with internalization of viral particles. The entry process was prevented by incubation of reconstituted cells with DHBV at 4 degrees C and by the addition of energy-depleting agents known to block DHBV entry into PDH. These results demonstrated that DCPD is a DHBV receptor. However, the lack of complete viral replication in DCPD-reconstituted cells suggested that additional factors are required for postentry events in immortalized cell lines.


title: p63: the phantom of the tumor suppressor.

text: The last twenty years of research into p53 function has revealed some fascinating discoveries into the orchestration of tumor suppressor pathways with a multitude of putative drug targets being investigated. However, it was not until 1998 that the ancestral mother of p53 was documented. The eldest evolutionary conserved homolog of the p53 family is known today as p63. Originally, it was thought p63 was another tumor suppressor that could function in a similar capacity to p53. However, elegant demonstrations of the divergent roles that p63 plays as a key transcriptional regulator of the proliferation and differentiation cascade in stratified epithelia are documented. These data link deltaNp63alpha to adult tissue stem cell regulation and possibly "cancer stem cells". p63 lacks mutation in cancer development, which is in stark contrast to the classically high mutation status of p53 in a large compendium of cancer types. Perhaps suggesting a selective preference for p53 mutation. Why is p63 rarely mutated despite being part of the same gene family? Interestingly, p63 is often over-expressed and amplified in cancer, thus revealing a paradox. Is p63 required to provide cancer cell populations with a selective advantage as much as a loss of p53 function by mutation? Has p53 been masking a "phantom" with promising features as a target for drug development? Can we exploit the biochemical know how gained from the mass of p53 research to further elucidate deltaNp63alpha gene function? In this review, we will summarise the emerging advances that are elucidating deltaNp63alpha as a promising drug target.


Inhibition of coronavirus MHV-A59 replication by proteinase inhibitors.


title: RNA recombination in a coronavirus: recombination between viral genomic RNA and transfected RNA fragments.

text: Mouse hepatitis virus (MHV), a coronavirus, has been shown to undergo a high frequency of RNA recombination both in tissue culture and in animal infection. So far, RNA recombination has been demonstrated only between genomic RNAs of two coinfecting viruses. To understand the mechanism of RNA recombination and to further explore the potential of RNA recombination, we studied whether recombination could occur between a replicating MHV RNA and transfected RNA fragments. We first used RNA fragments which represented the 5' end of genomic-sense sequences of MHV RNA for transfection. By using polymerase chain reaction amplification with two specific primers, we were able to detect recombinant RNAs which incorporated the transfected fragment into the 5' end of the viral RNA in the infected cells. Surprisingly, even the anti-genomic-sense RNA fragments complementary to the 5' end of MHV genomic RNA could also recombine with the MHV genomic RNAs. This observation suggests that RNA recombination can occur during both positive- and negative-strand RNA synthesis. Furthermore, the recombinant RNAs could be detected in the virion released from the infected cells even after several passages of virus in tissue culture cells, indicating that these recombinant RNAs represented functional virion RNAs. The crossover sites of these recombinants were detected throughout the transfected RNA fragments. However, when an RNA fragment with a nine-nucleotide (CUUUAUAAA) deletion immediately downstream of a pentanucleotide (UCUAA) repeat sequence in the leader RNA was transfected into MHV-infected cells, most of the recombinants between this RNA and the MHV genome contained crossover sites near this pentanucleotide repeat sequence. In contrast, when exogenous RNAs with the intact nine-nucleotide sequence were used in similar experiments, the crossover sites of recombinants in viral genomic RNA could be detected at more-downstream sites. This study demonstrated that recombination can occur between replicating MHV RNAs and RNA fragments which do not replicate, suggesting the potential of RNA recombination for genetic engineering.


docid: bkdbxbzi

title: Health imperatives in foreign policy: the case of Malaysia.

text: Malaysia's global, regional and bilateral international health relations are surveyed against the historical backdrop of the country's foreign policy. Malaysia has always participated in multilateral agencies, most notably the World Health Organization, as such agencies are part of the longstanding fabric of "good international citizenship". The threats of infectious diseases to human health and economic activity have caused an intensification and an organizational formalization of Malaysian health diplomacy, both regionally and bilaterally. Such diplomacy has also established a basis for developing a wider set of cooperative relationships that go beyond responding to the threat of pandemics. As Malaysia approaches "developed" status, its health sector is becoming increasingly integrated into the global economy through joint research and development ventures and transnational investment. At the same time, it will have the technological, financial and human resources to play an expanded altruistic role in global and regional health.


SARS and Carlo Urbani.


title: Probing for the human coronavirus OC43 in multiple sclerosis.

text: Tissues acquired at autopsy from four patients with MS and biopsies from one patient with a probable diagnosis of MS were probed for the presence of OC43 RNA. This human coronavirus was not detected.


title: Ecology and conservation: contributions to One Health.

text: Although One Health is widely promoted as a more effective approach towards human, animal and ecosystem health, the momentum is still driven largely by health professionals, predominantly from the veterinary sector. While few can doubt the merits of interdisciplinary One Health approaches to tackle complex health problems, operating across the disciplines still presents many challenges. This paper focuses on the contributions of partners from ecology and conservation to One Health approaches, and identifies four broad areas which could act as a focus for practical engagement and bring ecological and conservation objectives more to the forefront of the One Health agenda: i) developing initiatives with shared conservation and health objectives, particularly in and around protected areas and including programmes addressing human reproductive health and mental health; ii) broadening concepts of health to extend beyond indicators of disease to include the assessment of ecological impacts; iii) the integration of ecological and epidemiological monitoring systems within protected areas to support conservation management and wildlife disease surveillance; iv) building partnerships to bring conservation, health, development and animal welfare agencies together to combat threats to global biodiversity and health from the international trade in wildlife and wildlife products.


docid: khgnlkvg

title: Severe acute respiratory syndrome: a challenge for public health practice in Hong Kong.

text: Severe acute respiratory syndrome (SARS) is now a global challenge affecting more than 8000 patients in different continents. The dictum of public health practice especially for infectious disease is "prevention better than cure". It is especially true for SARS as the treatment strategies remain diverse and experimental. Maintaining a healthy and hygienic environment can be one of the effective public health measures to combat infectious disease. The major challenge is that some of the most important public health measures are to be taken outside the health sector. The community also needs to be strengthened and equipped with the health skills to promote better health and hygiene. There is also the need to create a supportive environment conducive to health for long term sustainability. The WHO approach of promoting health through setting approach would be one possible solution to face the challenge. This paper will describe some of the public health initiatives in Hong Kong through "setting approach" and "community development model" in helping the society fight against SARS. With the emergence of SARS, this might be the time to globalise public health medicine as an important component of medical practice.


title: Life sciences and biotechnology in China.

text: Life science and biotechnology have become a top priority in research and development in many countries as the world marches into the new century. China as a developing country with a 1.3 billion population and booming economy is actively meeting the challenge of a new era in this area of research. Owing to support from the government and the scientific community, and reform to improve the infrastructure, recent years have witnessed a rapid progress in some important fields of life science and biotechnology in China, such as genomics and protein sciences, neuroscience, systematics, super-hybrid rice research, stem cell and cloning technology, gene therapy and drug/vaccine development. The planned expansion and development of innovation in related sectors and the area of bioethics are described and discussed.


title: Prevalence of natural virus infections in laboratory mice and rats used in Canada.

text: Results of serological tests carried out over a period of 6 years to detect the presence of antibodies against 14 indigenous viruses in mice and rats used in 32 Canadian institutions are reported. Close to 20,000 individual sera were tested by the complement fixation or the hemagglutination inhibition technics. In order of mouse colony prevalence the six most common viruses present were pneumonia virus of mice, mouse hepatitis virus, rat virus, minute virus of mice, Sendai, and Theiler's mouse encephalomyelitis viruses. The most common viruses present in rat colonies were minute virus of mice, K virus, coronaviruses (rat coronavirus or sialodacryoadenitis virus), rat virus, H-1, pneumonia virus of mice, Theiler's mouse encephalomyelitis viruses, Sendai, and reovirus 3.


title: How hantaviruses modulate cellular pathways for efficient replication?

text: Hantaviruses are zoonotic category-A pathogens that cause highly fatal diseases in humans. The hantaviral genome encodes three viral proteins: RNA-dependent RNA polymerase (RdRp or L protein), nucleocapsid protein (N), and a glycoprotein precursor (GPC), which is post-translationally cleaved into two surface glycoproteins Gn and Gc. The cytoplasmic tail of Gn interferes with interferon signaling pathways. N is a multifunctional molecule that was shown to be involved in the transcription and translation of viral proteins. N binds to the host mRNA caps and protects the degradation of mRNA 5' termini, which are later snatched and used as primers by the viral RdRp during transcription initiation. N also seems to lure the host translation machinery for the preferential translation of viral transcripts. Moreover, N was shown to delay the induction of cellular apoptosis and facilitate the transport and localization of viral ribonucleoproteins (RNPs) by exploiting the cellular cytoskeleton and SUMOlyation machinery. Therefore, with their limited protein coding capacity, hantaviruses have evolved several strategies to modulate cellular pathways for their efficient replication.


Background paper. Mapping epitopes on coronavirus glycoproteins.


Identification of coronaviral conserved sequences and application to viral genome amplification.


title: Confocal Imaging and Tissue-Specific Fluorescent Probes for Real-Time In Vivo Immunohistochemistry. Proof of the Concept in a Gastric Lymph Node Metastasis Model.

text: BACKGROUND Tumor-specific fluorescent antibodies, which can be recognized at a cellular or tissue level using optical imaging such as confocal laser endomicroscopy (CLE), could provide a means for rapid and accurate tumor diagnosis and staging. The aim of this study was to evaluate the ability of CLE to detect the presence of tagged cells within lymph nodes in an original simulated metastatic model. MATERIALS AND METHODS A solution of indocyanine green containing a suspension of porcine hepatocytes, marked with carboxy-fluorescein-succinimidyl-ester (CFSE), was injected endoscopically in the gastric submucosa of 10 pigs. Fluorescence lymphography using a near-infrared laparoscope was used to identify sentinel and secondary drainage nodes. Additionally, a nonfluorescent gastric and a mesenteric node were identified. Every 5-10 min, those nodes were scanned using probe-based or needle-based CLE (pCLE or nCLE). Immunohistochemistry (IHC) using anti-cytokeratin 18 antibodies was subsequently performed to confirm the presence of hepatocytes in the lymph nodes. RESULTS A total of 36 lymph nodes were analyzed with both CLE probes. Hepatocyte penetration in lymph nodes, as assessed by repeated CLE scanning, took 10-40 min after submucosal injection. Concordance between CLE and IHC was 84 and 72 % for pCLE and nCLE, respectively. False negatives were partly due to incomplete CFSE labeling of hepatocytes, which could not be recognized by CLE, but were detected with IHC. CONCLUSIONS Real-time CLE analysis effectively recognized the presence in perigastric nodes of marked hepatic cells that had been injected endoscopically in the stomach. Validation studies on tumor-bearing animals using tumor-specific antibodies should be performed.


Correlating Hemodynamic Changes and Occlusion Time after Flow Diverter Treatment of Bilateral Large Internal Carotid Artery Aneurysms.


title: Factors impacting the acceptance of traceability in the food supply chain in the United States of America.

text: Global demands for increased food safety and quality assurance programmes, increased global competition, changing government rules and regulations, political and trade barriers, bioterrorism, and identity preservation requirements in global markets are affecting the world's food supply chain. To satisfy changing market demands, all suppliers in the food supply chain must adaptto these global issues. Total asset visibility must be maintained in production, in process, in storage, and in transit.


title: Transcriptional profiling of dysplastic lesions in K14-HPV16 transgenic mice using laser microdissection.

text: In the K14-HPV16 transgenic mouse model of human papillomavirus (HPV)-associated squamous cell cancers, HPV16 E6 and E7 oncogenes and E1 and E2 regulatory genes are driven by the K14 keratinocyte-specific promoter. HPV transcription varies within the different layers of the epithelium. The correlation between HPV transcription patterns and disease pathogenesis is not well understood. Understanding these patterns is critical to designing and testing new HPV-specific therapeutic strategies. We examined HPV gene expression in homogenous populations of cells microdissected from the stratum basale, stratum spinosum, and stratum corneum of lesions from the transgenic mice using PALM microlaser technology. RNA extracted from each cell layer was subjected to two-step gene-specific RT-PCR and real-time quantitative nested PCR. To ensure specific amplification of spliced transcripts, the primers used for real-time nested PCR spanned the splice sites. High levels of E2 were detected in the basal and supra-basal layers of hyperplastic and dysplastic lesions. E7 and E6* levels increased significantly over time in stratum basale and stratum spinosum. E6** was expressed at much lower levels. We showed that the transgenic mice express correctly spliced E2 transcripts and are suitable as a preclinical model to test a therapeutic strategy using transcriptional regulation by the E2 protein.


The Coronavirus Pandemic in Israel: Implications for Radiation Oncology Departments.


Scheme of RNA transcription in calicivirus-infected cells.


Intrathecal humoral immune response in coronavirus induced encephalomyelitis of Lewis and BN rats.


title: Comparison between unidirectional barbed and polyglactin 910 suture in vaginal cuff closure in patients undergoing total laparoscopic hysterectomy.

text: The aim of the study was to compare the efficacy and safety of barbed unidirectional vs. polyglactin 910 suture in vaginal cuff closure on patients submitted to total laparoscopic hysterectomy. From November 2011 until March 2012, a prolective, comparative, longitudinal, non-randomized study was performed on patients submitted to total laparoscopic hysterectomy (TLH). On entry, patients were assigned to two different groups, Group 1: Vaginal cuff closure with unidirectional #00 (n = 25) barbed suture, and Group 2: Vaginal cuff closure with polyglactin 910 #1 suture. The length of closure time and the frequency of dehiscence in vaginal vault were measured on the 14th postoperative day. We did not find significant differences between the groups on the general characteristic analysis. However, significant differences were found (P = .029) in the average vaginal cuff closure time (12.7 ± 3.1 min. for barbed suture group vs. 20.4 ± 7.1 min. for polyglactin 910 group). No dehiscence case was found in any group. Based on the results of this study, we can conclude that unidirectional barbed suture reduces the surgical time for vaginal cuff closure during total laparoscopic hysterectomy and doesn't increase the vaginal cuff dehiscence risk.


title: Duration of immunity in dogs after vaccination or naturally acquired infection.

text: This paper reviews current scientific information about the duration of immunity induced in dogs by infection or vaccination. It describes the shortcomings of the methods used to measure the immune responses of dogs, and explains the need for basic studies on the nature of protective humoral and cellular responses, and standardised assays for the long-term duration of immunity to pathogens other than rabies. The information is inadequate to warrant uniform recommendations on the ideal intervals for vaccination; each vaccine must be evaluated on the basis of its own merits and the characteristics of the disease it is intended to guard against.


title: Inhibiting severe acute respiratory syndrome-associated coronavirus by small interfering RNA.

text: OBJECTIVE To evaluate the effectiveness of small interfering RNA (siRNA) on inhibiting severe acute respiratory syndrome (SARS)-associated coronavirus replication, and to lay bases for the future clinical application of siRNA for the treatment of viral infectious diseases. METHODS Vero-E6 cells was transfected with siRNA before SARS virus infection, and the effectiveness of siRNA interference was evaluated by observing the cytopathic effect (CPE) on Vero-E6 cells. RESULTS Five pairs of siRNA showed ability to reduce CPE dose dependently, and two of them had the best effect. CONCLUSION siRNA may be effective in inhibiting SARS-associated coronavirus replication.


title: Reverse transcription loop-mediated isothermal amplification for the detection of Porcine reproductive and respiratory syndrome virus.

text: Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen of swine. The objective of the current study is to investigate the feasibility of using reverse transcription loop-mediated isothermal amplification (RT-LAMP) for the detection of PRRSV. The RT-LAMP is a recently described DNA amplification technique reported to be simple, inexpensive, fast, and accurate. The RT-LAMP reaction was set up using 2 sets of primers that were designed to detect North American and European strains of PRRSV and performed successfully in a simple heat block. The specificity of the amplified product was demonstrated by restriction analysis. The RT-LAMP was able to detect 5 different PRRSV isolates. However, the limit of detection ranged between 10(2) and 10(4) 50% tissue culture infective dose/ml. The RT-LAMP was further evaluated using serum samples from animals of known infection status. The ability of RT-LAMP to detect PRRSV in serum from acutely infected animals was evaluated with 114 serum samples from 18 experimentally inoculated boars. Forty-nine of these samples tested positive by RT-LAMP, while 94 were positive by reverse transcription polymerase chain reaction (RT-PCR). The diagnostic specificity, evaluated with 100 known negative serum samples, was estimated as 99%. The feasibility of RT-LAMP to detect PRRSV was demonstrated in the current study. The RT-LAMP reaction could be performed in just 1 hr with a simple and inexpensive heat block. However, the sensitivity of this technique was significantly lower than that of RT-PCR.


title: [Primary investigation on the changing mode of plasma specific IgG antibody in SARS patients and their physicians and nurses].

text: OBJECTIVES To primarily investigate the changing mode of anti-SARS coronavirus IgG antibody in clinically diagnosed SARS patients and the possibility of subclinical infection in physicians and nurses through close association with SARS patients. METHODS The plasma levels of anti-SARS coronavirus IgG antibody of 57 normal subjects, 127 physicians and nurses worked in SARS wards for one month and 73 SARS patients with different course of SARS were measured by enzyme linked immunosorbent assay. RESULTS Plasma anti-SARS coronavirus IgG antibody was not detected in normal subjects and the clinical personnel. After 0-7 days, 8-10 days, 11-14 days, 15-20 days of onset of disease, the positive rates were 0.33%, 52%, 86% respectively, and the general positive rate was 61%. CONCLUSION The specificity and sensitivity of ELISA to detect plasma anti-SARS IgG antibody were satisfactory. Cases with positive reaction could be diagnosed as patients already infected by the virus. The specific IgG antibody didn't emerge in some patients in the early stage of the disease, and the negative results didn't indicate that they were not infected. And follow-up investigation should be made in those patients. Unlike common epidemic infectious diseases, SARS probably hadn't the potentiality of subclinical infection.


title: Mouse hepatitis virus receptors: more than a single carcinoembryonic antigen.

text: Mouse hepatitis virus (MHV), a murine coronavirus, has been shown to utilize carcinoembryonic antigen (CEA) as the receptor. We have demonstrated that MHV can utilize a different isoform of CEA, which is an alternatively spliced gene product that is expressed in different tissues, as a receptor. Furthermore, the CEA molecules from a resistant mouse strain (SJL) have different sequences and yet serve as functional viral receptors. Thus, MHV can use more than a single type of CEA molecule as the receptor. We have also shown that some mouse cell lines express functional CEA molecules and yet are resistant to infections by certain MHV strains. Biochemical studies of the infected cells indicate that MHV infections in these cell lines are blocked at the steps of virus entry. We conclude that MHV entry requires additional cellular factors other than CEA, the viral receptor. The significance of viral receptors and the additional cellular factors in regulating viral tropism is discussed.


title: Mannose-binding lectin (MBL) facilitates opsonophagocytosis of yeasts but not of bacteria despite MBL binding.

text: Mannose-binding lectin (MBL) is a serum protein of the innate immune system. After binding to a microorganism, MBL in complex with MBL-associated serine proteases activates the complement system, resulting in cleavage of complement factor C3. Cleaved C3 on the surface of the microorganism mediates opsonization for clearance, but the impact of MBL on subsequent phagocytosis has not been widely studied. We investigated the role of MBL in complement activation and phagocytosis of various bacteria and yeast species by flow cytometry. We measured both the C3 deposition during serum opsonization of fluorescent-labeled microorganisms as well as subsequent uptake of these microorganisms by human neutrophils. In MBL-deficient sera, a consistently decreased C3 deposition on both zymosan and Candida albicans was found and a reduced phagocytosis by neutrophils that was restored by exogenous MBL. This indicates that the lectin pathway of complement activation is important for the opsonophagocytosis of yeasts. In contrast, the C1q-dependent classical pathway dominated in the opsonization and phagocytosis of Staphylococcus aureus, Streptococcus pneumoniae, and Escherichia coli, whereas no effect of MBL was found. Both the lectin and the classical pathway of complement activation were highly amplified by the alternative route for opsonophagocytosis by neutrophils of yeast as well as microbial species. In summary, our data demonstrate that yeast species are preferentially opsonized and subsequently phagocytosed via activation of the lectin pathway of complement, whereas the uptake of bacterial strains was found to be largely MBL independent.


title: RNA recombination of coronaviruses: localization of neutralizing epitopes and neuropathogenic determinants on the carboxyl terminus of peplomers.

text: Murine coronaviruses undergo RNA recombination at a very high frequency. We have obtained a series of recombinant viruses using neutralizing monoclonal antibodies in conjunction with temperature-sensitive markers. All of the recombinants obtained have a crossover within gene C, which encodes the peplomer protein of the virus. The genetic structure of these recombinants suggests that the antigenic regions recognized by these neutralizing monoclonal antibodies are localized on the carboxyl-terminal one-third of the peplomer protein. Since the two monoclonal antibodies used are also associated with the critical determinants of virus neuropathogenicity, we conclude that both the neutralizing antibody binding sites and determinants of pathogenicity are localized at the carboxyl-terminal one-third of the peplomer. The variation of crossover sites in different recombinant viruses also allowed precise mapping of additional antigenic sites. RNA recombination thus presents a powerful genetic tool, and the carboxyl-terminal localization of the biological functions of peplomers suggests a distinct conformation of these viral membrane proteins.


title: [Detection of SARS-CoV and RNA on aerosol samples from SARS-patients admitted to hospital].

text: OBJECTIVE To assess the risk of aerosol transmission in severe acute respiratory syndrome (SARS) patients admitted to Hospital through testing the air samples. METHODS Air samples were collected from 7 wards and 1 balcony of the Hospital, 3 times a day for 3 continuous days, using bioaerosol sampler type FA-2. Bioaerosol particles were then washed down from the samples by serum-free Dulbecco's Modified Eagle Medium (DMEM) culture medium. Nested-reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the N protein gene of the SARS associated coronavirus (SARS-CoV) from these washing solutions. The residual solutions were inoculated into prepared cell cultures to isolate live virus. The positive samples were then identified by indirect immunofluorescence assay and sequence analysis of the PCR products. RESULTS Positive rates of RT-PCR test on air samples were 29.03% in the wards and 20.0% in balcony respectively. Results from sequential analysis showed that the homology of amplified cDNA fragments to previously known SARS-CoV stains was 98%. A strain of live pathogen was isolated from one of the 36 samples. The isolate could cause typical cytopathic effects, similar to those SARS-CoV on Vero-E6 cells and the effects could be stably passed. Indirect immunofluorescence assay showed positive from serum of a SARS patient. CONCLUSION SARS-CoV existed in the air hospital, where SARS patients were admitted to, but the activity of SARS-CoV in air samples was rather low. SARS patients could still shed SARS-CoV even during the recovery phase. Potential possibility of aerosol transmission might exist within 1 meter square area around SARS patients.


Influenza A/H1N1 from a pig perspective.


title: A retrospective study of 74 laparoscopic repairs of abdominal incisional hernias.

text: INTRODUCTION A laparoscopic procedure is used more and more frequently to treat incisional hernia with the potential benefits of shorter hospitalisation and a decrease in postoperative pain. The purpose of this retrospective study was to analyse the results of the laparoscopic treatment of incisional hernia at our institution and to identify potential risk factors for recurrence. METHODS The medical data (pre-operative, peri-operative, and postoperative) of patients who received a laparoscopic repair of their incisional hernia between January 2003 and February 2007 were recorded. The follow-up was based on a retrospective analysis of the information found in the patients' medical records. RESULTS Seventy-four laparoscopic interventions were performed on 71 patients. Polyester implants with an average size of 412.16 cm2 were used to cover the hernia. The mean operative time was 76.8 +/- 55.6 min (range, 20 to 295 min) and the mean duration of post-operative hospitalisation was 3.75 +/- 2.3 days (range, 2 to 12 days). One breach in the small intestine (1.4%) (sutured with 3/0 silk thread) and 1 conversion to laparotomy (1.4%) for a voluminous incisional hernia occurred during surgery. The post-operative morbidity was 8.2%, the rate of long-term complications was 27%, and 13 recurrences (including 3 with complications) were noted (17.6%) during a mean follow-up of 13 months. There was no postoperative mortality. Recurrences were linked to the use of large meshes corresponding to large incisional hernia diameter (p < 0.05). CONCLUSION Although the morbidity/mortality rates are acceptable, technical improvements must be found to reduce the recurrence rate, in particular for large incisional hernias.


The guardian.


COVID-19 Pandemic - Nephrology Experiences.