title: [The expression and activity detection of a variant N protein of SARS-CoV].

text: AIM To construct an expression vector pGEX-2T/N, and to express the fusion protein consisting of N protein of SARS-CoV and GST in E.coli. METHODS The N region gene of SARS-CoV was cloned by RT-PCR. The expression vector was constructed by DNA recombination. The recombinant plasmid was transformed into E.coli BL21(DE3). The expression of the fusion protein was detected by Western blot. RESULTS (1) As compared with the sequences in GenBank, 20 bp were deleted in DNA sequence of the cloned N protein. (2) The fusion protein GST-N was soluble. Western blot analysis showed that the reaction of GST-N to anti-SARS-CoV sera was positive. CONCLUSION The pGEX-2T/N has been constructed and expressed in the form of fusion protein GST-N successfully, which lays the foundation for further study of SARS-CoV N protein.


title: An office-based approach to influenza: clinical diagnosis and laboratory testing.

text: Vaccination is the primary measure for preventing morbidity and mortality from influenza. During the influenza season, family physicians must distinguish influenza from the common cold and other flu-like illnesses. Signs and symptoms of influenza include abrupt onset of fever, severe myalgias, anorexia, sore throat, headache, cough, and malaise. Clinical diagnosis can be difficult or nonspecific when patients have other symptoms (e.g., stuffy nose, sneezing, cough, sore throat) that can be caused by various respiratory viruses or bacterial pathogens. Family physicians can improve diagnostic accuracy by being aware of the epidemiology of influenza. During outbreaks of influenza, commercially available rapid assays can be used to identify type A and B viruses. On average, rapid in-office tests are more than 70 percent sensitive and 90 percent specific for viral antigens. The assays vary in complexity, specificity, sensitivity, time to obtain results, specimen analyzed, and cost. The results of rapid viral tests can guide treatment decisions.


title: Nonassociative explanations of behavioral changes induced by central cholinergic drugs.

text: Studies concerning the effects of cholinergic stimulants and blockers on habituation, alternation, classical and instrumental conditioning, differentiation, and discrimination are summarized and discussed. This analysis suggests that many results originally ascribed to associative changes can be more economically accounted for by nonassociative factors inherent in the experiments. Furthermore, the data show that neither one of the more economical nonassuciative explanations at either the sensory, the drive, or the response level can by itself account for the complex interactions observed. The studies dealing with drug effects in lesioned animals, and those using intracerebral treatments, point out several sites of action of muscarinic agonists and antagonists in the septum, the hippocampus, the amygdala, the striatum, the hypothalamus, midline thalamic nuclei, and the reticular formation. Therefore, the provisional conclusion must be that central muscarinic systems subserve several functions, ranging from the modulation of consummatory responses to the control of motor-act differentiation. Both the drug and the lesion studies show not only that the fronto-limbic system has a considerable role in the latter process, but also that the localization of motor-act differentiation under a given drive depends both on the category of the cue, and on the category of the response. Some comparisons between the effects of muscarinic agonists and blockers and those of other drugs indicate that the alteration of sensory-motor relations caused by the former can be separated from (i) the motor hyperactivity caused by amphetamine, which is relatively less dependent on sensory factors; (ii) the selective (response enhancements caused by the indole hallucinogen LSD25, which appear to be amenable to a disinhibition of drive; (iii) the attenuation of punishment suppression by sedative and antianxiety agents (benzodiazepines and subhypnotic doses of barbiturates); and (iv) the hyperarousal or hyperreactivity syndrome induced by compounds which deplete central 5-hydroixytryphamine stores, such as panachlorophenylalanine.


title: The possible use of combined electrical impedance and ultrasound velocity measurements for the non-invasive measurement of temperature during mild hyperthermia.

text: This paper explores the possibility of using combined measurements of electrical impedance and changes in ultrasound time of flight for determining deep body temperature during mild hyperthermia. Simultaneous electrical impedance spectra (1 kHz-1024 kHz) and ultrasound time-of-flight measurements were made on layered sheep liver and fat tissue samples as the temperature was increased from 30-50 °C. The change in propagation velocity for 100% fat and 100% liver samples was found to vary linearly with temperature and the temperature coefficient of the time-of-flight was shown to vary linearly with the % fat in the sample (0.009% °C-1%-1). Tetrapolar impedance measurements normalized to 8 kHz were shown to have a small sensitivity to temperature for both liver (0.001% °C-1 ≤ 45 °C) and fat (0.002% °C-1 ≤ 512 kHz) and the best linear correlation between the normalized impedance and the % fat in the sample was found at 256 kHz (gradient 0.026%-1, r2 = 0.65). A bootstrap analysis on 15 layered tissue samples evaluated using the normalized impedance at 256 kHz to determine the % fat in the sample and the temperature coefficient of the time of flight to determine the temperature. The results showed differences (including some large differences) between the predicted and measured temperatures and an error evaluation identified the possible origins of these.


title: Rapid and sensitive detection of severe acute respiratory syndrome coronavirus by rolling circle amplification.

text: The severe acute respiratory syndrome (SARS) epidemic of 2003 was responsible for 774 deaths and caused significant economic damage worldwide. Since July 2003, a number of SARS cases have occurred in China, raising the possibility of future epidemics. We describe here a rapid, sensitive, and highly efficient assay for the detection of SARS coronavirus (SARS-CoV) in cultured material and a small number (n = 7) of clinical samples. Using rolling circle amplification (RCA), we were able to achieve sensitive detection levels of SARS-CoV RNA in both solid and liquid phases. The main advantage of RCA is that it can be performed under isothermal conditions with minimal reagents and avoids the generation of false-positive results, a problem that is frequently encountered in PCR-based assays. Furthermore, the RCA technology provides a faster, more sensitive, and economical option to currently available PCR-based methods.


title: Effect of 2 Bedding Materials on Ammonia Levels in Individually Ventilated Cages.

text: This study sought to identify an optimal rodent bedding and cage-change interval to establish standard procedures for the IVC in our rodent vivarium. Disposable cages were prefilled with either corncob or α-cellulose bedding and were used to house 2 adult Sprague-Dawley rats (experimental condition) or contained no animals (control). Rats were observed and intracage ammonia levels measured daily for 21 d. Intracage ammonia accumulation became significant by day 8 in experimental cages containing α-cellulose bedding, whereas experimental cages containing corncob bedding did not reach detectable levels of ammonia until day 14. In all 3 experimental cages containing α-cellulose, ammonia exceeded 100 ppm (our maximum acceptable limit) by day 11. Two experimental corncob cages required changing at days 16 and 17, whereas the remaining cage containing corncob bedding lasted the entire 21 d without reaching the 100-ppm ammonia threshold. These data suggests that corncob bedding provides nearly twice the service life of α-cellulose bedding in the IVC system.


Prevalence of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus infection in Korean pigs.


title: Infection with cytotoxic T-lymphocyte escape mutants results in increased mortality and growth retardation in mice infected with a neurotropic coronavirus.

text: C57BL/6 mice infected with mouse hepatitis virus strain JHM (MHV-JHM) develop a chronic demyelinating encephalomyelitis several weeks after inoculation. Previously, we showed that mutations in the immunodominant CD8 T-cell epitope (S-510-518) could be detected in nearly all samples of RNA and virus isolated from these mice. These mutations abrogated recognition by T cells harvested from the central nervous systems of infected mice in direct ex vivo cytotoxicity assays. These results suggested that cytotoxic T-lymphocyte (CTL) escape mutants contributed to virus amplification and the development of clinical disease in mice infected with wild-type virus. In the present study, the importance of these mutations was further evaluated by infecting naive mice with MHV-JHM variants isolated from infected mice and in which epitope S-510-518 was mutated. Compared to mice infected with wild-type virus, variant virus-infected animals showed higher mortality and morbidity manifested by decreased weight gain and neurological signs. Although a delay in the kinetics of virus clearance has been demonstrated in previous studies of CTL escape mutants, this is the first illustration of significant changes in clinical disease resulting from infection with viruses able to evade the CD8 T-cell immune response.


Proposed new International Health Regulations.


title: Monoclonal antibody analysis of neutralization and antibody-dependent enhancement of feline infectious peritonitis virus.

text: Fifty-four monoclonal antibodies (MAbs) to feline infectious peritonitis virus (FIPV) were characterized according to protein specificity, immunoglobulin subclass, virus neutralization, reactivity with different coronaviruses, and ability to induce antibody-dependent enhancement (ADE) of FIPV infection in vitro. The MAbs were found to be specific for one of three structural proteins of FIPV. A total of 47 MAbs were specific for the 205-kDa spike protein (S), 3 MAbs were specific for the 45-kDa nucleocapsid protein (N), and 4 MAbs were specific for the 26- to 28-kDa membrane protein (M). The S-specific MAbs showed various degrees of cross-reactivity with strains of FIPV, feline enteric coronavirus, canine coronavirus, and porcine transmissible gastroenteritis virus. Nineteen S-specific MAbs neutralized FIPV. A total of 15 of the neutralizing MAbs induced ADE, and all but 1 were of the immunoglobulin G2a subclass. The remaining four neutralizing MAbs that did not induce ADE were of the immunoglobulin G1 subclass. Two S-specific MAbs induced ADE but were nonneutralizing. None of the N- or M-specific MAbs was neutralizing or induced ADE. On the basis of the reactivity patterns of the MAbs with FIPV and related coronaviruses, it was concluded that there is a minimum of five neutralizing sites on S. In most instances, neutralizing MAbs were able to induce ADE, demonstrating a direct relationship between neutralization and enhancement. The difference in immunoglobulin subclass between neutralizing MAbs that induced ADE and those that did not induce ADE suggests that there may be a restriction in the immunoglobulin subclasses capable of mediating ADE.


title: Identification and primary structure of the gene encoding the Berne virus nucleocapsid protein.

text: The nucleotide sequence of the nucleocapsid (N) protein gene of Berne virus (BEV; proposed family Toroviridae) was determined from two independent clones of a cDNA library. From the deduced amino acid sequence a basic protein of 18.3K was predicted. In vitro transcription and translation, followed by immunoprecipitation, were used to identify the gene. The identification was confirmed by metabolic labelling, using the knowledge that cysteine residues are absent from the amino acid sequence of the N protein. Smaller N-related polypeptides encountered in BEV-infected cell lysates were shown to be probable products of aberrant translation, due to initiation on AUG codons further downstream in the N protein gene.


title: Protection of mice from lethal coronavirus MHV-A59 infection by monoclonal affinity-purified spike glycoprotein.

text: Numerous studies have provided indirect evidence that the spike glycoprotein of coronaviruses (E2 or S) bears determinants for pathogenesis and the induction of protective immunity. In order to directly evaluate its immunogenicity, the E2 glycoprotein of the murine hepatitis virus, strain A59, was purified by immunoaffinity chromatography. High titers of neutralizing and fusion inhibiting antibodies were induced in mice vaccinated with purified E2/S in Freund's adjuvant, which were protected from an intracerebral challenge with 10 LD50 of MHV-A59. This study provides a direct demonstration of the importance of the coronavirus spike glycoprotein in the induction of a protective immune response.


title: [Male violence against heterosexual partners: facts and challenges. Review of Mexican literature].

text: Violence against women by partners (VAWP) has been systematically studied in Mexico in the past 15 years. The aim of this paper is to reflect on the contributions on this topic in Mexico. The discussion focuses on the magnitude of this phenomenon, stressing its theoretical, methodological, and technical implications; the relationship between violence and a variety of factors; and the outcomes of VAWP on women themselves. The manuscript features the main contributions from quantitative and qualitative studies.


Inflammation and coagulation in the cardiovascular system: the contribution of influenza.


title: Comparison of the di- and trinucleotide frequencies from the genomes of nine different coronaviruses.

text: As an alternative to protein alignments for the comparison of sequences, the reiterations of mono- di- and trinucleotide frequencies were used for the comparison of coronavirus sequences. The relative abundance of the di- and trinucleotide frequencies within the 3' part from nine coronavirus genomes were determined. The patterns of dinucleotide frequencies and the trinucleotide frequencies showed some common features for all coronaviruses but also differences between the groups formerly defined on the base of antigenic relatedness. The normalised dinucleotide frequencies were further used to calculate the distances between coronavirus sequences. Based on the dinucleotide frequency distances, coronaviruses can be divided into two groups which roughly reflect the taxonomic groups. In this kind of evaluation, however, IBV occupies a position different to the one that it would take based on most protein sequence comparisons. Based on similarities within coding sequences and antigenic properties IBV occupies a place outside of both groups. Based on the dinucleotide frequencies IBV gained a position in between of the TGEV-related and the MHV-clustered coronaviruses.


title: Influenza and other actue respiratory diseases in the Czech Socialist Republic, 1969-1974.

text: Since 1969, the incidence of acute respiratory diseases (ARD) in the Czech Socialist Republic of Czechoslovakia has been monitored by a special programme based on reports from 85 district epidemiological centres. In this paper, the incidence of ARD in three age groups, together with the incidence of complications and death rates, are presented for each season during the period 1969-1974. The significance of epidemiological observations and laboratory investigations relating to influenza and other ARD agents, such as parainfluenza viruses, adenoviruses, rhinoviruses, RS virus, coronaviruses, and Mycoplasma pneumoniae, is discussed.


title: Spatial clustering of SARS in Hong Kong.

text: 1. Geographic information system (GIS) can be applied during an acute infectious disease outbreak to reveal new geospatial information in addition to standard field epidemiological analyses. 2. When applied in real time during the onset and evolution of an epidemic, GIS can monitor and enhance understanding of the transmission dynamics of an infectious agent, thereby facilitating the design, implementation and evaluation of potential intervention strategies.


title: Feline infectious peritonitis in a cat presented because of papular skin lesions.

text: A 19-week-old neutered male domestic shorthair cat was examined because of multiple raised pruritic skin lesions along the dorsal head and back. Histopathology of biopsies of the lesions detected nodular pyogranulomatous dermatitis with vasculitis and necrosis, leading to a suspicion of feline infectious peritonitis (FIP). Postmortem examination revealed gross lesions consistent with FIP. Histopathologic lesions and positive immunohistochemical staining for feline coronavirus in multiple tissues, including the skin, confirmed the diagnosis of FIP. The current case was similar to previous cases, except for the initial presentation with cutaneous lesions and no other clinical signs, which had not been reported previously.


title: Virally activated CD8 T cells home to Mycobacterium bovis BCG-induced granulomas but enhance antimycobacterial protection only in immunodeficient mice.

text: The effect of secondary infections on CD4 T-cell-regulated chronic granulomatous inflammation is not well understood. Here, we have investigated the effect of an acute viral infection on the cellular composition and bacterial protection in Mycobacterium bovis strain bacille Calmette-Guérin (BCG)-induced granulomas using an immunocompetent and a partially immunodeficient murine model. Acute lymphocytic choriomeningitis virus (LCMV) coinfection of C57BL/6 mice led to substantial accumulation of gamma interferon (IFN-gamma)-producing LCMV-specific T cells in liver granulomas and increased local IFN-gamma. Despite traffic of activated T cells that resulted in a CD8 T-cell-dominated granuloma, the BCG liver organ load was unaltered from control levels. In OT-1 T-cell-receptor (TCR) transgenic mice, ovalbumin (OVA) immunization or LCMV coinfection of BCG-infected mice induced CD8 T-cell-dominated granulomas containing large numbers of non-BCG-specific activated T cells. The higher baseline BCG organ load in this CD8 TCR transgenic animal allowed us to demonstrate that OVA immunization and LCMV coinfection increased anti-BCG protection. The bacterial load remained substantially higher than in mice with a more complete TCR repertoire. Overall, the present study suggests that peripherally activated CD8 T cells can be recruited to chronic inflammatory sites, but their contribution to protective immunity is limited to conditions of underlying immunodeficiency.


title: Utility of the over-the-scope-clip system for treating a large esophageal perforation.

text: We report here a case of esophageal perforation made by an endoscope while treating cicatrical stenosis that developed after wide circumferential dissection of superficial esophageal carcinoma. Perforation closure with a conventional endoclip was difficult as the perforation was large and the surrounding tissue was fragile as a result of steroids administration for stenosis prevention. To avoid surgical intervention, we employed the over-the-scope-clip system and successfully closed the perforation. The favorable outcome suggests the utility of the over-the-scope-clip system for closing perforations when conventional methods are ineffective.


title: Structure-function relationships of the viral RNA-dependent RNA polymerase: fidelity, replication speed, and initiation mechanism determined by a residue in the ribose-binding pocket.

text: Studies of the RNA-dependent RNA polymerase (RdRp) from poliovirus (PV), 3Dpol, have shown that Asn-297 permits this enzyme to distinguish ribose from 2'-deoxyribose. All animal RNA viruses have Asn at the structurally homologous position of their polymerases, suggesting a conserved function for this residue. However, all prokaryotic RNA viruses have Glu at this position. In the presence of Mg2+, the apparent affinity of Glu-297 3Dpol for 2'-deoxyribonucleotides was decreased by 6-fold relative to wild type without a substantial difference in the fidelity of 2'-dNMP incorporation. The fidelity of ribonucleotide misincorporation for Glu-297 3Dpol was reduced by 14-fold relative to wild type. A 4- to 11-fold reduction in the rate of ribonucleotide incorporation was observed. Glu-297 PV was unable to grow in HeLa cells due to a replication defect equivalent to that observed for a mutant PV encoding an inactive polymerase. Evaluation of the protein-(VPg)-primed initiation reaction showed that only half of the Glu-297 3Dpol initiation complexes were capable of producing VPg-pUpU product and that the overall yield of uridylylated VPg products was reduced by 20-fold relative to wild-type enzyme, a circumstance attributable to a reduced affinity for UTP. These studies identify the first RdRp derivative with a mutator phenotype and provide a mechanistic basis for the elevated mutation frequency of RNA phage relative to animal RNA viruses observed in culture. Although protein-primed initiation and RNA-primed elongation complexes employ the same polymerase active site, the functional differences reported here imply significant structural differences between these complexes.


title: [Prevalence and seasonal distribution of respiratory viruses in patients with acute respiratory tract infections, 2002-2014].

text: The aim of this study was to investigate the prevalence and seasonal distribution of respiratory viruses in pediatric and adult outpatients and inpatients who were admitted to hospital with the symptoms of upper and lower respiratory tract infections, during a 12-year period. A total of 5102 clinical samples (4372 nasopharyngeal swabs, 316 bronchoalveolar lavages, 219 transtracheal aspirates, 163 nasopharyngeal aspirates, 20 sputum, 10 nasal swabs) examined in our laboratory between January 1st 2002 and July 17th 2014, were evaluated retrospectively. Of the specimens, 1107 (21.7%) were obtained from outpatients and 3995 (78.3%) from hospitalized patients. Of the patients, 2851 (55.9%) were male and 2251 (44.1%) were female, while 1233 (24.2%) were adults and 3869 (75.8%) were children (age range: 1 day - 93 years; median: 3 years). Respiratory samples were investigated for the presence of respiratory syncytial virus (RSV), influenza virus type A and B (INF-A, INF-B), adenovirus (AdV), parainfluenza viruses (PIV types 1-4), human rhinoviruses (HRV), human coronaviruses (HCoV), human metapneumovirus (HMPV) and human bocavirus (HBoV). All specimens were tested by both direct immunofluorescence antibody (DFA) and shell vial cell culture (SVCC) methods. In DFA assay the samples were initially screened by fluorescent-labeled polyclonal antibodies, and the positive ones were typed by using monoclonal antibodies (Light Diagnostics, Merck Millipore, USA). In SVCC, HEp-2, MDCK, A-549 and Vero cell lines were used for the isolation of viruses. In addition to these methods, real-time multiplex PCR methods (RealAccurate®, Respiratory RT PCR, PathoFinder, Netherlands and Seeplex® RV15 ACE Detection, Seegene, South Korea) were used for the detection of respiratory viruses in samples (n= 2104) obtained from 2007 to 2014. Respiratory viruses were detected in a total of 1705 (33.4%) patients, of them 967 (19%) were male and 738 (14.4%) were female. Three hundred and eighteen (18.6%) of the 1705 patients were infected with multiple respiratory viruses. The most frequently observed co-infections were RSV+INF-A (40/318; 12.6%), and RSV+PIV (33/318; 10.4%). The rate of positivity for the respiratory viruses in pediatric and adult groups were 35.4% (1369/3869) and 27.3% (336/1233), respectively (p< 0.000). The most frequently detected virus in pediatric group was RSV (336/1369; 24.5%), followed by influenza viruses (314/1369; 22.9%), PIV (197/1369; 14.4%), HRV (118/1369; 8.6%), AdV (75/1369; 5.5%) and the others (49/1369; 3.6%). On the other hand the most frequently detected virus in adult group was influenza viruses (181/336; 53.8%) followed by AdV (37/336; 11%), RSV (24/336; 7.1%), PIV (24/336; 7.1%), HRV (23/336; 6.8%) and the others (9/336; 2.7%). The rate of multiple virus infections in pediatric and adult groups were 7.2% (280/3869) and 3% (38/1233), respectively. Most of the coinfections (280/318; 88%) were detected in children. Respiratory viruses were detected positive in 40.2% (445/1107) of outpatients, and in 31.5% (1260/3995) of inpatients (p< 0.000). The most frequent viruses detected in pediatric outpatients and inpatients were HRV and RSV, respectively, while influenza viruses were the first in line among both adult outpatients and inpatients. During the study period, a PIV-3 outbreak (n= 96) have emerged between December 2004-April 2005, and an influenza A (H1N1)pdm09 outbreak (n= 207) between November 2009-January 2010. When the seasonal distribution was considered, the isolation rates of 1705 respiratory viruses in winter, spring, summer and autumn were 44.4%, 27%, 8.3% and 20.3%, respectively. RSV was most frequently detected from December to March, influenza viruses from November to March, HRV from December to June, and mixed infections from January to February. In conclusion, the data of our study obtained in about 12-year period indicated that the prevalence of respiratory viruses in acute respiratory infections is 33.4%, and they typically active during the months of winter and early spring in our region.


title: Human coronavirus 229E: receptor binding domain and neutralization by soluble receptor at 37 degrees C.

text: Truncated human coronavirus HCoV-229E spike glycoproteins containing amino acids 407 to 547 bound to purified, soluble virus receptor, human aminopeptidase N (hAPN). Soluble hAPN neutralized the infectivity of HCoV-229E virions at 37 degrees C, but not 4 degrees C. Binding of hAPN may therefore trigger conformational changes in the viral spike protein at 37 degrees C that facilitate virus entry.


title: Diversity of influenza-like illness etiology in Polish Armed Forces in influenza epidemic season.

text: The aim of this study was to conduct an epidemiological and laboratory surveillance of Influenza-Like Illnesses (ILI) in Polish Armed Forces, civilian military personnel and their families in 2011/2012 epidemic season, under the United States Department of Defense-Global Emerging Infections Surveillance and Response System (DoD-GEIS). ILI incidence data were analyzed in relation to age, gender, patient category as well as pathogen patterns. Multiple viral, bacterial and viral-bacterial co-infections were identified. Nose and throat swabs of active duty soldiers in the homeland country and in the NATO peacekeeping forces KFOR (Kosovo Force), as well as members of their families were tested for the presence of viral and bacterial pathogens. From October 2011 to May 2012, 416 specimens from ILI symptoms patients were collected and analyzed for the presence of viral and bacterial pathogens. Among viruses, coronavirus was the most commonly detected. In the case of bacterial infections, the most common pathogen was Staphylococcus aureus.


title: Bladder cancer. Clinical practice guidelines in oncology.

text: Urothelial tumors represent a spectrum of diseases with a range of prognosis. After a diagnosis is established at any point within the urothelial tract, the patient remains at risk for developing a new lesion at a different or the same location and at a similar or more advanced stage. Continued monitoring for recurrence is an essential part of management, because most recurrences are superficial and can be managed endoscopically. Within each category of disease, more refined methods to determine prognosis and guide management, based on molecular staging, are under development with the goal of optimizing the individual patient's likelihood of cure and chance for organ preservation. For patients with more extensive disease, newer treatments typically involve combined-modality approaches, using recently developed surgical procedures, or three-dimensional treatment planning for more precise delivery of radiation therapy. Although these are not appropriate in all cases, they do offer the promise of an improved quality of life and prolonged survival. Finally, within the category of metastatic disease, a number of new agents have been identified that appear to be superior to those currently considered to be standard therapies. Experts believe, therefore, that the treatment of urothelial tumors will evolve rapidly over the next few years, with improved outcomes for patients at all stages of disease.


title: Development and evaluation of reverse transcription-loop-mediated isothermal amplification assay for rapid and real-time detection of Japanese encephalitis virus.

text: The standardization and validation of a one-step, single-tube accelerated quantitative reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay is reported for rapid and real-time detection of Japanese encephalitis virus (JEV). The RT-LAMP assay reported in this study is very simple and rapid; the amplification can be obtained in 30 min under isothermal conditions at 63 degrees C by employing a set of six primers targeting the E gene of JEV. The RT-LAMP assay demonstrated exceptionally higher sensitivity compared to that of RT-PCR, with a detection limit of 0.1 PFU. The specificities of the selected primer sets were established by cross-reactivity studies with other closely related members of the JEV serocomplex as well as by evaluation of healthy human volunteers. The comparative evaluation of the RT-LAMP assay for clinical diagnosis with a limited number of patient cerebrospinal fluid samples revealed 85% concordance with conventional RT-PCR, with a sensitivity and a specificity of 100% and 86%, respectively. The concentration of virus in most of the clinical samples was 10(2) to 10(5) PFU/ml, as determined from the standard curve based on the time of positivity in the samples. In addition, the monitoring of gene amplification can also be visualized with the naked eye by using SYBR green I fluorescent dye. Thus, due to easy operation without a requirement of sophisticated equipment and skilled personnel, the RT-LAMP assay reported here is a valuable tool for the rapid and real-time detection of JEV not only by well-equipped laboratories but also by peripheral diagnostic laboratories with limited financial resources in developing countries.


title: Qigong practice among chronically ill patients during the SARS outbreak.

text: AIMS AND OBJECTIVES This research aimed at exploring the motivations and experiences of chronically ill participants practising qigong during the severe acute respiratory syndrome outbreak in Hong Kong. BACKGROUND Although biomedicine is the mainstream medical system in Hong Kong, many people employ complementary and alternative medicine in dealing with their chronic health problems. Practising qigong is one of the most popular forms of complementary and alternative medicine used among chronically ill patients. Little is known about the experiences of the chronically ill patients practising qigong in the severe acute respiratory syndrome outbreak and even less is known how this practice is related to the social context of Hong Kong during the outbreak. DESIGN AND METHODS Qualitative methods using participant-observation from three qigong classes in Hong Kong and in-depth semi-structured interviews with 30 participants were employed. Content analysis and discourse analysis were used to identify major themes of the data. RESULTS Both the underlying and trigger motivations could motivate these chronically ill participants to practise qigong. Legitimacy of qigong in health maintenance, deterioration of health and unpleasant experiences in biomedical treatment on their chronic illnesses served as the underlying motivations. Stigmatization of and discrimination against the chronically ill and the sense of searching coping strategy during the severe acute respiratory syndrome outbreak further motivated their practice. CONCLUSION To the participants, practising qigong not only could strengthen their health, but was also a coping strategy for them to regain an active control and the sense of security in their health and enabled them to overcome the social stigmatization and discrimination during the outbreak. RELEVANCE TO CLINICAL PRACTICE. Searching for emotional support and a more active role in health conditions could be seen from the chronically ill participants. Understanding and paying more attention to the particular needs of the chronically ill patients can enable health-care professionals to provide better care and support for the chronically ill during an epidemic crisis.


title: Construction of a mouse hepatitis virus recombinant expressing a foreign gene.

text: The genome of the coronavirus mouse hepatitis virus (MHV) contains genes which have been shown to be nonessential for viral replication and which could, in principle, be used as sites for the introduction of foreign sequences. We have inserted heterologous genetic material into gene 4 of MHV in order (i) to test the applicability of targeted RNA recombination for site-directed mutagenesis of the MHV genome upstream of the N gene; (ii) to develop further genetic tools for mutagenesis of structural genes other than N; and (iii) to examine the feasibility of using MHV as an expression vector. A DI-like donor RNA vector containing the MHV S gene and all genes distal to S was constructed. Initially, a derivative of this was used to insert a 19-nucleotide tag into the start of ORF 4a of MHV-A59 using the N gene deletion mutant A1b4 as the recipient virus. Subsequently, the entire gene for the green fluorescent protein (GFP) was inserted in place of gene 4. This heterologous gene was shown to be expressed by recombinant viruses but not at levels sufficient to allow detection of fluorescence of viral plaques. Northern blot analysis of transcripts of GFP recombinants showed the expected displacement of the mobility, relative to those of wild-type, of all subgenomic mRNAs larger than mRNA5. An unexpected result of the Northern analysis was the observation that GFP recombinants also produced an RNA species the same size as that of wild-type mRNA4. RT-PCR analysis of the 5' end of this species revealed that it was actually a collection of mRNAs originating from a cluster of 10 different sites, none of which possessed a canonical intergenic sequence. The finding of these aberrant mRNAs, all of nearly the same size as wild-type mRNA4, suggests that long range structure of the MHV genome can sometimes be the sole determinant of the site of initiation of transcription.


DNA vaccines and apoptosis: to kill or not to kill?


title: Supplementation with 3 compositionally different tocotrienol supplements does not improve cardiovascular disease risk factors in men and women with hypercholesterolemia.

text: BACKGROUND Tocotrienols have been reported to lower LDL-cholesterol and fasting glucose concentrations and to have potent antioxidant effects, but the results are contradictory. OBJECTIVE The objective was to study the relative effect of tocotrienol supplements of different compositions (mixed alpha- plus gamma-, high gamma-, or P25-complex tocotrienol) on blood lipids, fasting blood glucose, and the excretion of 8-iso-prostaglandin F(2alpha), a measure of oxidative stress, in healthy hypercholesterolemic men and women. DESIGN This was a double-blind, randomized, parallel-design study in which subjects (n = 67 men and women) consumed 1 of 3 commercially available tocotrienol supplements or a safflower oil placebo for 28 d. Blood and urine samples were obtained before and after the 28-d supplementation phase for analysis of fasting blood lipids, glucose, tocotrienols and tocopherols, and 8-iso-prostaglandin F(2alpha). RESULTS Overall, serum tocotrienols were increased in subjects who consumed tocotrienols, which showed that the putatively active components were absorbed. No significant differences in mean lipid or glucose concentrations were observed among the 4 treatment groups at the end of the 28-d supplementation phase. However, when the values were expressed as a percentage change from the concentrations during the presupplementation run-in phase, LDL cholesterol increased slightly (7 +/- 2%) but significantly (P < 0.05) in the group consuming the mixed alpha- plus gamma-tocotrienol supplement when compared with LDL cholesterol in the group consuming the P25-complex tocotrienol. Neither mean concentrations nor the percentage change in 8-iso-prostaglandin F(2alpha) differed significantly among treatments. CONCLUSION Supplementation with 200 mg tocotrienols/d from 3 commercially available sources has no beneficial effect on key cardiovascular disease risk factors in highly compliant adults with elevated blood lipid concentrations.


title: Multimeric humanized varicella-zoster virus antibody fragments to gH neutralize virus while monomeric fragments do not.

text: Murine monoclonal antibody 206 (MAb mu206) binds to gH, the varicella-zoster virus (VZV) fusogen, neutralizing the virus in vitro in the absence of complement and inhibiting cell-to-cell spread and egress of VZV in cultured cells. We have humanized this antibody to generate MAb hu206 by complementarity determining region grafting. MAb hu206 retained binding and in vitro neutralizing activity, as well as cross-reactivity with ten different VZV strains. Single-chain antibody fragments (scAb) derived from MAb hu206 were produced in Escherichia coli. These scAb retained the binding properties of the whole antibody. However, monomeric scAb exhibited markedly reduced neutralizing activity compared to the bivalent parental MAb hu206. Shortening the peptide linker joining the V(H) to the V(kappa) domain from 14 to 5 or even 0 residues encouraged multimerization and increased neutralizing efficacy. The fact that Fab fragments enzymatically generated from whole MAb hu206 lost their neutralizing potency lent support to the proposal that valency is important for VZV neutralization at this epitope.


title: Molecular detection and identification of influenza viruses by oligonucleotide microarray hybridization.

text: Microarrays of virus-specific oligonucleotides may provide a method of screening samples for the presence or absence of a large variety of viruses simultaneously. Influenza viruses are ideal for evaluating such microarrays because of their genetic and host diversity, and the availability of an extensive sequence database. A collection of 476 influenza virus-specific oligonucleotides was spotted onto glass slides as probes. Viral RNAs were reverse transcribed and amplified by PCR, and the products were labeled with cyanine dyes. The presence of viruses and their identities were determined by hybridization. The fluorescence intensities of oligonucleotide spots were highly reproducible within each slide and satisfactorily proportional between experiments. However, the intensities of probe spots completely complementary to target sequences varied from background to saturation. The variations did not correlate with base composition, nucleotide sequence, or internal secondary structures. Therefore, thresholds for determining whether hybridization to a spot should be judged as positive were assigned individually. Considering only positive spots from probes predicted to be monospecific for influenza virus species, subtype, host source, or gene segment, this method made correct identifications at the species, hemagglutinin subtype, and gene segment levels. Monospecific neuraminidase (NA) subtype probes were insufficiently diverse to allow confident NA subtype assignment. Incorporating positive spots from polyspecific probes into the identification scheme gave similar results. Overall, the results demonstrate the potential of microarray-based oligonucleotide hybridization for multiple virus detection.


title: Hendra and nipah infection: pathology, models and potential therapies.

text: The Paramyxoviridae family comprises of several genera that contain emerging or re-emerging threats for human and animal health with no real specific effective treatment available. Hendra and Nipah virus are members of a newly identified genus of emerging paramyxoviruses, Henipavirus. Since their discovery in the 1990s, henipaviruses outbreaks have been associated with high economic and public health threat potential. When compared to other paramyxoviruses, henipaviruses appear to have unique characteristics. Henipaviruses are zoonotic paramyxoviruses with a broader tropism than most other paramyxoviruses, and can cause severe acute encephalitis with unique features among viral encephalitides. There are currently no approved effective prophylactic or therapeutic treatments for henipavirus infections. Although ribavirin was empirically used and seemed beneficial during the biggest outbreak caused by one of these viruses, the Nipah virus, its efficacy is disputed in light of its lack of efficacy in several animal models of henipavirus infection. Nevertheless, because of its highly pathogenic nature, much effort has been spent in developing anti-henipavirus therapeutics. In this review we describe the unique features of henipavirus infections and the different strategies and animal models that have been developed so far in order to identify and test potential drugs to prevent or treat henipavirus infections. Some of these components have the potential to be broad-spectrum antivirals as they target effectors of viral pathogenecity common to other viruses. We will focus on small molecules or biologics, rather than vaccine strategies, that have been developed as anti-henipaviral therapeutics.


Immunopathogenesis of demyelination induced by MHV-4.


Orogastric administration of crushed darunavir tablets for a critically ill patient.


Saint Louis University School of Medicine and Vaccine Center Mobilize for COVID-19 Pandemic.


Using BRCA1 to treat cancer.


title: A cross-sectional validation study of using NERDS and STONEES to assess bacterial burden.

text: All chronic wounds are colonized by micro-organisms. Although the presence of bacteria is not necessarily harmful, and may be beneficial in some instances, accurate evaluation of wound-related bacterial damage and infection is crucial. A cross-sectional validation study involving 112 patients was conducted to estimate the specificity and sensitivity of clinical assessment variables individually and in combination to determine the presence and quantity of bacteria in the wound. The average age of study participants was 66 years (range 33 to 95 years) and most had leg (44) and foot (68) ulcers of approximately 6 months' duration. Wounds were evaluated using a mnemonic developed to evaluate the presence or absence of clinical signs of critical colonization (NERDS) or infection (STONEES) and results compared to semi-quantitative swab cultures. Wounds with debris, increased exudate, and friable tissue were found to be five times more likely to have scant or light bacterial growth; whereas, wounds with elevated temperature were eight times more likely to have moderate or heavy bacterial growth. When combining any three clinical signs, the sensitivity was 73.3% for scant or light and 90% for moderate and heavy bacterial growth and the specificity was 80.5% and 69.4%, respectively. Considering the importance of this clinical diagnosis, studies to examine the predictive validity of these assessment variables and culture results are warranted.


title: Rat strains differ in susceptibility to Ureaplasma parvum-induced urinary tract infection and struvite stone formation.

text: Individuals with struvite uroliths are susceptible to recurrent urinary tract infections (UTI), sepsis, and renal disease. Unfortunately, little is known about the host-specific factors that predispose to this disease. In order to develop a rodent model that can address this problem, we inoculated female Fischer 344 (F344), Lewis (LEW), Sprague-Dawley (SD), and Wistar (WIS) rats with a host-adapted strain of Ureaplasma parvum. Animals were necropsied at 2 weeks postinoculation; 100% of F344, 42% of SD, 10% of LEW, and 10% of WIS rats remained infected. Severe bladder lesions and struvite calculi were seen in 64% of F344 rats; in other rat strains, bladder lesions were mild or absent. F344 rats with struvite uroliths had the highest urinary levels of proinflammatory cytokines, such as GRO/KC, interleukin-1alpha (IL-1alpha), and IL-1beta. F344 rats without struvite stones at necropsy had milder bladder lesions and significantly lower urinary levels of proinflammatory cytokines but a more prominent inflammatory response than did other rat strains. Based on our results, struvite stone formation is linked to a robust inflammatory response that does not resolve UTI but instead promotes damage to surrounding tissues.


title: mRNA helicase activity of the ribosome.

text: Most mRNAs contain secondary structure, yet their codons must be in single-stranded form to be translated. Until now, no helicase activity has been identified which could account for the ability of ribosomes to translate through downstream mRNA secondary structure. Using an oligonucleotide displacement assay, together with a stepwise in vitro translation system made up of purified components, we show that ribosomes are able to disrupt downstream helices, including a perfect 27 base pair helix of predicted T(m) = 70 degrees . Using helices of different lengths and registers, the helicase active site can be localized to the middle of the downstream tunnel, between the head and shoulder of the 30S subunit. Mutation of residues in proteins S3 and S4 that line the entry to the tunnel impairs helicase activity. We conclude that the ribosome itself is an mRNA helicase and that proteins S3 and S4 may play a role in its processivity.


title: [Detection of the mRNA expression of human angiotensin-converting enzyme 2 as a SARS coronavirus functional receptor in human femoral head].

text: OBJECTIVE To investigate the mRNA expression of severe acute respiratory syndrome-associated coronavirus (SARS-COV) functional receptor, angiotensin-converting enzyme 2 (ACE2), in human femoral head and conjunctiva, and explore the possible entry route of SARS-COV in human femoral head. METHODS ACE2 mRNA in human femoral head was detected by nested RT-PCR with human beta actin gene as the positive control. RESULTS The mRNA of human beta actin gene could be amplified efficiently in all the tissue samples. The mRNA of human ACE2 was expressed efficiently in the normal lung tissue, but not in the cartilage and cancellous bone under the weight-bearing area of the femoral head. CONCLUSION SARS-COV can not infect the femoral head tissue and lead to avascular necrosis of the femoral head directly by the spike glycoprotein, and mechanism of the virus for causing avascular necrosis needs further investigation.


title: Lymphopenia in severe acute respiratory syndrome: a summary on its frequency.

text: Severe acute respiratory syndrome (SARS) is a new human infectious disease caused by coronavirus. The first world outbreak occurs in 2002, starting from China and then thorough many countries. Abnormal hematological variables were common among patients with SARS. Concerning lymphocyte disorder, there were some previous studies reported the low lymphocyte in the patients with SARS. Here, the author performs a further analysis to document the frequency of the lymphopenia in SARS by the way of metanalysis study. Five available reports concerning the prevalence of lymphopenia in SARS among different populations were selected for further analysis in this study. Overall 637 SARS patients were retrospectively analyzed and 492 cases presented lymphopenia. According to this study, the overall prevalence rate of lymphopenia in SARS is 77.2 %. No significant correlation between the population ethnicity and the prevalence rate was detected in this study (P > 0.05). Here, the lymphopenia can be documented as an important characteristic of blood picture in the patients with SARS or it can indicate that "SARS is a viral-induced lymphopenia disease".


title: A panel of optimized primers and positive-control DNAs for PCR detection of common biological contaminants in mouse cell lines and tissue samples.

text: PCR-based testing for infectious agents in mouse cell lines and tissues has recently been developed as an alternative to the traditional MAP test. One drawback to currently available PCR-based assays is the lack of appropriate positive controls for PCR detection of the infectious agents. When negative samples are the norm and positive controls are absent, it is very difficult to feel confident detecting infectious agents. To alleviate this problem, the authors developed a panel of primers and positive-control DNA plasmids that enable rapid testing of biological samples, such as cell lines, tissues, or animal sera, for presence of the infectious agents most damaging to mouse colonies.


title: Intracellular complexes of viral spike and cellular receptor accumulate during cytopathic murine coronavirus infections.

text: Murine hepatitis virus (MHV) infections exhibit remarkable variability in cytopathology, ranging from acutely cytolytic to essentially asymptomatic levels. In this report, we assess the role of the MHV receptor (MHVR) in controlling this variable virus-induced cytopathology. We developed human (HeLa) cell lines in which the MHVR was produced in a regulated fashion by placing MHVR cDNA under the control of an inducible promoter. Depending on the extent of induction, MHVR levels ranged from less than approximately 1,500 molecules per cell (designated R(lo)) to approximately 300,000 molecules per cell (designated R(hi)). Throughout this range, the otherwise MHV-resistant HeLa cells were rendered susceptible to infection. However, infection in the R(lo) cells occurred without any overt evidence of cytopathology, while the corresponding R(hi) cells died within 14 h after infection. When the HeLa-MHVR cells were infected with vaccinia virus recombinants encoding MHV spike (S) proteins, the R(hi) cells succumbed within 12 h postinfection; R(lo) cells infected in parallel were intact, as judged by trypan blue exclusion. This acute cytopathology was not due solely to syncytium formation between the cells producing S and MHVR, because fusion-blocking antiviral antibodies did not prevent it. These findings raised the possibility of an intracellular interaction between S and MHVR in the acute cell death. Indeed, we identified intracellular complexes of S and MHVR via coimmunoprecipitation of endoglycosidase H-sensitive forms of the two proteins. We suggest that MHV infections can become acutely cytopathic once these intracellular complexes rise above a critical threshold level.


Hormone receptors in breast cancer.


title: Noninvasive positive pressure ventilation in the intensive care unit: a concise review.

text: OBJECTIVE To critically assess available high-level clinical studies regarding use of noninvasive positive pressure ventilation in varied intensive care unit settings. DATA SOURCE Search of pertinent articles within Ovid MEDLINE from 1975 to 2005, CINAHL from 1982 to 2005, EMBASE from 1988 to 2005, and Web of Science from 1993 to 2005. STUDY SELECTION Randomized, controlled clinical trials and cohort studies and observational studies the authors consider important or novel. DATA EXTRACTION/SYNTHESIS Performed equally by both authors with the use of an Excel data spreadsheet. CONCLUSION There is abundant level I evidence supporting the use of noninvasive positive pressure ventilation in such critical care settings as acute hypercapnic respiratory failure, particularly related to chronic obstructive pulmonary disease, and acute cardiogenic pulmonary edema. We also report on other clinical scenarios in which the data may be somewhat less compelling, but evidence favors a noninvasive positive pressure ventilation trial. Some well designed studies suggest that noninvasive positive pressure ventilation is not an appropriate intervention for patients who have failed endotracheal extubation.


Rechallenging immunological memory.


title: The association of titers to bovine coronavirus with treatment for bovine respiratory disease and weight gain in feedlot calves.

text: The association between bovine respiratory disease (BRD) and antibody titers to bovine coronavirus (BCV) was studied in 604 calves (19 different groups in 4 different feedlots from 2 provinces). Almost all calves had antibody titers on arrival in the Alberta feedlot and 82% of the calves had an antibody titer on arrival at the Ontario feedlots; titers in calves in Alberta were almost twice as high as those in calves in Ontario. The incidence of infection, in the first mo after arrival as judged by seroconversion, ranged from 61% to 100%; titer increases were much greater in calves in Ontario feedlots. Titer variables were not significantly related to BRD, except on a within-group basis (group was a confounding variable for BCV-BRD associations). Given control of group effects, calves with an antibody titer on arrival appeared to be protected against BRD for the first 28 d in the feedlot, and the association was reasonably linear over the range of titers. Each titer unit on arrival decreased the risk of BRD by about 0.8x (odds ratio). Titer change was not strongly related to the risk of BRD and the relationship was not linear over the range of titer changes. Titer change was strongly and negatively correlated with titer on arrival, and titer change was not significantly related to BRD in the presence of arrival titers. Arrival titer retained its relationship with BRD in the presence of titer data for other putative pathogens. Each higher unit of titer to BCV on arrival increased the 28-day weight gain (controlling for group, initial weight and the occurrence of BRD) by slightly more than 1 kg. Titer change was associated with decreased weight gain, when initial titer was not in the model. The lack of a linear or multivariable association between BCV titer change and BRD, and weight gain, may indicate that BCV is not a major pathogen; or, its lack of significance may merely be due to its strong correlation with arrival titer. Given the associations found in this study, particularly the interprovincial differences in arrival titers, more and different approaches to studying the possible effects of BCV on BRD are in order.


Planetary health: what is it and what should doctors do?


title: Expression of reporter genes from the defective RNA CD-61 of the coronavirus infectious bronchitis virus.

text: The defective RNA (D-RNA) CD-61, derived from the Beaudette strain of the avian coronavirus infectious bronchitis virus (IBV), was used as an RNA vector for the expression of two reporter genes, luciferase and chloramphenicol acetyltransferase (CAT). D-RNAs expressing the CAT gene were demonstrated to be capable of producing CAT protein in a helper-dependent expression system to about 1.6 microgram per 10(6) cells. The reporter genes were expressed from two different sites within the CD-61 sequence and expression was not affected by interruption of the CD-61-specific ORF. Expression of the reporter genes was under the control of a transcription-associated sequence (TAS) derived from the Beaudette gene 5, normally used for the transcription of IBV subgenomic mRNA 5. The Beaudette gene 5 TAS is composed of two tandem repeats of the IBV canonical consensus sequence involved in the acquisition of a leader sequence during the discontinuous transcription of IBV subgenomic mRNAs. It is demonstrated that only one canonical sequence is required for expression of mRNA 5 or for the expression of an mRNA from a D-RNA and that either sequence can function as an acceptor site for acquisition of the leader sequence.


title: Lyme disease--United States, 2003-2005.

text: Lyme disease is caused by the spirochete Borrelia burgdorferi and is transmitted to humans by the bite of infected blacklegged ticks (Ixodes spp.). Early manifestations of infection include fever, headache, fatigue, and a characteristic skin rash called erythema migrans. Left untreated, late manifestations involving the joints, heart, and nervous system can occur. A Healthy People 2010 objective (14-8) is to reduce the annual incidence of Lyme disease to 9.7 new cases per 100,000 population in 10 reference states where the disease is endemic (Connecticut, Delaware, Maryland, Massachusetts, Minnesota, New Jersey, New York, Pennsylvania, Rhode Island, and Wisconsin). This report summarizes surveillance data for 64,382 Lyme disease cases reported to CDC during 2003-2005, of which 59,770 cases (93%) were reported from the 10 reference states. The average annual rate in these 10 reference states for the 3-year period (29.2 cases per 100,000 population) was approximately three times the Healthy People 2010 target. Persons living in Lyme disease--endemic areas can take steps to reduce their risk for infection, including daily self-examination for ticks, selective use of acaricides and tick repellents, use of landscaping practices that reduce tick populations in yards and play areas, and avoidance of tick-infested areas.


title: Serosurvey of viral infections in free-ranging Namibian cheetahs (Acinonyx jubatus).

text: Cheetahs (Acinonyx jubatus) in captivity have unusually high morbidity and mortality from infectious diseases, a trait that could be an outcome of population homogeneity or the immunomodulating effects of chronic stress. Free-ranging Namibian cheetahs share ancestry with captive cheetahs, but their susceptibility to infectious diseases has not been investigated. The largest remaining population of free-ranging cheetahs resides on Namibian farmlands, where they share habitat with domestic dogs and cats known to carry viruses that affect cheetah health. To assess the extent to which free-ranging cheetahs are exposed to feline and canine viruses, sera from 81 free-ranging cheetahs sampled between 1992 and 1998 were evaluated for antibodies against canine distemper virus (CDV), feline coronavirus (feline infectious peritonitis virus; FCoV/ FIPV), feline herpesvirus 1 (FHV1), feline panleukopenia virus (FPV), feline immunodeficiency virus (FIV), and feline calicivirus (FCV) and for feline leukemia virus (FeLV) antigens. Antibodies against CDV, FCoV/FIPV, FHV1, FPV, and FCV were detected in 24, 29, 12, 48, and 65% of the free-ranging population, respectively, although no evidence of viral disease was present in any animal at the time of sample collection. Neither FIV antibodies nor FeLV antigens were present in any free-ranging cheetah tested. Temporal variation in FCoV/FIPV seroprevalence during the study period suggested that this virus is not endemic in the free-ranging population. Antibodies against CDV were detected in cheetahs of all ages sampled between 1995 and 1998, suggesting the occurrence of an epidemic in Namibia during the time when CDV swept through other parts of sub-Saharan Africa. This evidence in free-ranging Namibian cheetahs of exposure to viruses that cause severe disease in captive cheetahs should direct future guidelines for translocations, including quarantine of seropositive cheetahs and preventing contact between cheetahs and domestic pets.


title: Could SARS-CoV-2/COVID-19 simply fade away?

text: not available.


title: The complete genome sequence of the major component of a mild citrus tristeza virus isolate.

text: The genome of the Spanish mild isolate T385 of citrus tristeza virus (CTV) was completely sequenced and compared with the genomes of the severe isolates T36 (Florida), VT (Israel) and SY568 (California). The genome of T385 was 19,259 nt in length, 37 nt shorter than the genome of T36, and 33 and 10 nt longer than those of VT and SY568, respectively, but their organization was identical. T385 had mean nucleotide identities of 81.3, 89.3 and 94% with T36, VT and SY568, respectively. The 3' UTR had over 97% identity in all isolates, whereas the 5' UTR of T385 had 67% identity with VT, 66.3% with SY568 and only 42.5% with T36. In the coding regions, the nucleotide differences between T385 and VT were evenly distributed along the genome (around 90% identity); this was not observed between T385 and the other isolates. T385 and T36 had nucleotide identities around 90% in the eight 3'-terminal ORFs of the genome, but only 72.3% in ORF 1a, a divergence pattern similar to that reported previously for T36 and VT. T385 and SY568 had nucleotide identities close to 90% in the 5'- and 3'-terminal regions of the genome, whereas the central region had over 99% identity. Our data suggest that the central region in the SY568 genome results from RNA recombination between two CTV genomes, one of which was almost identical to T385.


title: Genetic determinants of Sindbis virus mosquito infection are associated with a highly conserved alphavirus and flavivirus envelope sequence.

text: Wild-type Sindbis virus (SINV) strain MRE16 efficiently infects Aedes aegypti midgut epithelial cells (MEC), but laboratory-derived neurovirulent SINV strain TE/5'2J infects MEC poorly. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of MRE16 and TE/5'2J differ at 60 residue sites. To identify the genetic determinants of MEC infection of MRE16, the TE/5'2J virus genome was altered to contain either domain chimeras or more focused nucleotide substitutions of MRE16. The growth patterns of derived viruses in cell culture were determined, as were the midgut infection rates (MIR) in A. aegypti mosquitoes. The results showed that substitutions of MRE16 E2 aa 95 to 96 and 116 to 119 into the TE/5'2J virus increased MIR both independently and in combination with each other. In addition, a unique PPF/.GDS amino acid motif was located between these two sites that was found to be a highly conserved sequence among alphaviruses and flaviviruses but not other arboviruses.


title: Human rhinovirus infection in children with cystic fibrosis.

text: Nasopharyngeal swabs obtained from 78 pediatric patients with cystic fibrosis (CF), including 47 with acute pulmonary exacerbation and 31 in a stable clinical condition, were evaluated for 17 respiratory viruses. Human rhinovirus (HRV) was the most frequently detected virus in patients with pulmonary exacerbation and in those who were clinically stable (21.3% vs. 12.9%; P = 0.52). HRV-A was the main RV detected in patients with pulmonary exacerbations. However, no prevalence of particular HRV-A subtypes was found. This study highlights that RV is frequently found in the respiratory secretions of patients with CF and the impact of HRV-A appears higher than that of the other HRV types in patients with pulmonary exacerbations.


title: Differences among confirmed and not-confirmed COVID-19 patients at "D.Cotugno" hospital, Naples (Italy): what we learned from first suspected cases?

text: Clinical presentation of COVID-19 is common to other respiratory infections. We compared the characteristics at hospital admission of confirmed and not-confirmed COVID-19 patients, in the early phase of the epidemic. Thirty-seven suspected patients were enrolled, and COVID-19 was confirmed in 17. Confirmed patients are older, have more frequently contact with confirmed cases. Distinctive clinical characteristics among COVID-19 were the grand-glass opacities at CT scan, and a pO2/FiO2 ratio less than 250. In not-confirmed group, Influenza represented the most frequent alternative diagnosis. This study contributes to highlight the characteristics to consider at hospital admission in order to promptly suspect COVID-19.


Review Team.


title: Epidemiological study of enteric coronavirus excretion by an enzyme-linked immunosorbent assay.

text: The enzyme-linked immunosorbent assay with a blocking test was used to determine the prevalence of enteric coronavirus excretion among the population of Bangkok and Khon Kaen province. The results indicated that coronaviruses were present in fecal samples from both children and adults with or without diarrhea. It is interesting that the viruses were more frequently observed in the apparently healthy individuals in Bangkok than in the patients with diarrhea whereas the excretion rates were approximately alike in both groups in Khon Kaen province. Among children aged less than 5 years (with or without diarrhea) the viruses were more frequently observed in stools from young children (1-5 years old) than from the newborn (less than 1-year-old) particularly those living in the Bang Khen slum area and in the rural Khon Kaen community. The rate of coronavirus excretion did not increase with age and some apparently healthy individuals continued to excrete the viruses for several months. The results also suggested that chronic parasitic infection associated with low socioeconomic status might influence the excretion of enteric coronaviruses in the feces.


The resurgence of West Nile virus.


title: [SARS: diagnosis, therapy, and especially prevention].

text: The main purpose of this review is to analyze some aspects of the severe acute respiratory syndrome, SARS, in order to obtain useful data to suggest preventive actions to reduce the spreading of the disease. Many elements have been examined to reach some conclusions and to allow an updated discussion. Surgical masks protect more the patient than the caregiver. Simple or double surgical masks may be useful, as double gloving protects the hands of the surgical personnel against percutaneous transmission of HIV eventually present in contaminated blood. The frequent substitution of the external masks with a new one will improve the filtering activity against droplets produced by cough or sneezes of the patient. The use of respiratory masks may be suggested in hospitals or in restricted ventilated areas where, even if coronavirus variant is considered an environmental contaminant more than a respiratory risk, droplets nuclei may persist in the air and add consistent dangers to the heath-care givers. Considering that large and medium droplets may infect floors and surfaces, in addition to gloves, gowns, masks and eyes protection, the available list of viral and bacterial factors implicated in SARS ethiology suggests a better hand antisepsis using frequently the alcohol based gels (containing an high percentage of emollients substances), if available. A liquid soap with triclosan can also be used, if the health-care workers compliance to hand washing increases, as expected in this explosive situation. On the basis of the results of some experimental data, the environmental disinfection may be effected with ethyl alcohol 70% in water. Disinfection of floors or larger surfaces may be obtained with chlorine compounds solutions, after an accurate pre-cleaning. When corrosion, bleaching or gas production have to be avoided, chlorine compounds may be substituted by phenolic detergent disinfectants.


title: Molecular epidemiology of bovine coronavirus on the basis of comparative analyses of the S gene.

text: Bovine coronavirus (BCoV), a group 2 member of the genus Coronavirus in the family Coronaviridae, is an important pathogen in cattle worldwide. It causes diarrhea in adult animals (winter dysentery), as well as enteric and respiratory diseases in calves. The annual occurrence of BCoV epidemics in Sweden and Denmark led to this investigation, with the aim to deepen the knowledge of BCoV epidemiology at the molecular level. A total of 43 samples from outbreaks in both countries were used for PCR amplification and direct sequencing of a 624-nucleotide fragment of the BCoV S gene. Sequence comparison and phylogenetic studies were performed. The results showed (i) identical sequences from different animals in the same herds and from paired nasal and fecal samples, suggesting a dominant virus circulating in each herd at a given time; (ii) sequence differences among four outbreaks in different years in the same herd, indicating new introduction of virus; (iii) identical sequences in four different Danish herds in samples obtained within 2 months, implying virus transmission between herds; and (iv) that at least two different virus strains were involved in the outbreaks of BCoV in Denmark during the spring of 2003. This study presents molecular data of BCoV infections that will contribute to an increased understanding of BCoV epidemiology in cattle populations.


Learn from influenza epidemiology.


title: Defensive applications of gene transfer technology in the face of bioterrorism: DNA-based vaccines and immune targeting.

text: Gene transfer involves the introduction of an engineered gene into a person's cells with the expectation that the protein expressed from the gene will produce a therapeutic benefit. Strategies based on this principle have led to the approval of > 600 clinical trials and enrollment of approximately 3500 subjects worldwide in attempts to treat diseases ranging from cancer to AIDS to cystic fibrosis. While gene therapy has met with limited success and still has many hurdles to overcome before it sees wide application, it may be useful as a defensive strategy against bioterrorism agents including infectious microbes and toxins. Although many defensive strategies are possible, immunological strategies are currently the most developed and are being actively applied to the development of strategies against several of the most virulent potential bio-weapons. While most of these strategies are not yet ready for human application, DNA-based vaccines appear to be among the most promising in the fight against bioterrorism.


title: Tagging Recombinant Proteins to Enhance Solubility and Aid Purification.

text: Protein fusion technology has had a major impact on the efficient production and purification of individual recombinant proteins. The use of genetically engineered affinity and solubility-enhancing polypeptide "tags" has increased greatly in recent years and there now exists a considerable repertoire of these that can be used to solve issues related to the expression, stability, solubility, folding, and purification of their fusion partner. In the case of large-scale proteomic studies, the development of purification procedures tailored to individual proteins is not practicable, and affinity tags have therefore become indispensable tools for structural and functional proteomic initiatives that involve the expression of many proteins in parallel. Here, the rationale and applications of a range of established and more recently developed solubility-enhancing and affinity tags is described.


title: Protective effect of Houttuynia cordata extract on bleomycin-induced pulmonary fibrosis in rats.

text: The present study aimed to examine the antioxidant properties of Houttuynia cordata (HC) and its protective effect on bleomycin-induced pulmonary fibrosis in rats. Results showed that aqueous extract of HC exhibited a different magnitude of antioxidant activities in all model systems tested. Although HC showed weaker free radical scavenging and xanthine oxidase inhibitory activity than vitamin E, its anti-lipid peroxidation activity in rat liver homogenate was close to that of vitamin E. In animal studies, HC significantly decreased the levels of superoxide dismutase, malondialdehyde, hydroxyproline, interferon-gamma, and tumor necrosis factor-alpha. However, an increase in the concentration of catalase was noted in the bronchoalveolar lavage fluid. HC also remarkably improved the morphological appearance of the lung of bleomycin-treated rats. These results suggest that HC possesses a protective effect against bleomycin-induced pulmonary fibrosis. Interestingly, this protective effect was more pronounced than that of vitamin E. In conclusion, the protective effect of HC on pulmonary fibrosis could be partly associated with the reduction of oxidative damage caused by bleomycin.


title: Identification of immunodominant epitopes on the membrane protein of the severe acute respiratory syndrome-associated coronavirus.

text: Similar to other coronaviruses, the membrane (M) protein of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is a major transmembrane glycoprotein with multiple biological functions. To date, limited information is available about its antigenic properties. In this study, we identified two major immunodominant epitopes on the M protein located in the extreme N-terminal region (residues 1 to 31) and the interior C-terminal region (residues 132 to 161), respectively, by Pepscan analyses against convalescent-phase sera from SARS patients and antisera from virus-immunized mice and rabbits. Synthetic peptides M1-31 derived from the N-terminal epitope and M132-161 derived from the C-terminal epitope were highly reactive with all of the convalescent-phase sera from 40 SARS patients but not with 30 control serum samples from healthy blood donors, suggesting their potential application for serologic diagnosis of SARS. We showed that both peptides (M1-31 and M132-161) were able to induce high titers of antibody responses in the immunized rabbits, highlighting their antigenicity and immunogenicity. These findings provide important information for developing SARS diagnostics and vaccines.


title: Highlights and conclusions from the technical consultative meeting on novel coronavirus infection, Cairo, Egypt, 14-16 January 2013.

text: The emergence of a novel strain of coronavirus in the Arabian Peninsula raised a global health concern in 2012, partly because the majority of human infections were fatal and partly due to its presumed animal origin. An urgent meeting of scientific and public health experts was convened by WHO in January 2013 in view of the limited knowledge available on the epidemiological and natural history of infection with this novel virus. The meeting reviewed current evidence and identified critical knowledge gaps to improve better understanding of the public health risk associated with the virus so as to improve preparedness and to safeguard and protect global health.


title: Interpretation of diagnostic laboratory tests for severe acute respiratory syndrome: the Toronto experience.

text: BACKGROUND An outbreak of severe acute respiratory syndrome (SARS) began in Canada in February 2003. The initial diagnosis of SARS was based on clinical and epidemiological criteria. During the outbreak, molecular and serologic tests for the SARS-associated coronavirus (SARS-CoV) became available. However, without a "gold standard," it was impossible to determine the usefulness of these tests. We describe how these tests were used during the first phase of the SARS outbreak in Toronto and offer some recommendations that may be useful if SARS returns. METHODS We examined the results of all diagnostic laboratory tests used in 117 patients admitted to hospitals in Toronto who met the Health Canada criteria for suspect or probable SARS. Focusing on tests for SARS-CoV, we attempted to determine the optimal specimen types and timing of specimen collection. RESULTS Diagnostic test results for SARS-CoV were available for 110 of the 117 patients. SARS-CoV was detected by means of reverse-transcriptase polymerase chain reaction (RT-PCR) in at least one specimen in 59 (54.1%) of 109 patients. Serologic test results of convalescent samples were positive in 50 (96.2%) of 52 patients for whom paired serum samples were collected during the acute and convalescent phases of the illness. Of the 110 patients, 78 (70.9%) had specimens that tested positive by means of RT-PCR, serologic testing or both methods. The proportion of RT-PCR test results that were positive was similar between patients who met the criteria for suspect SARS (50.8%, 95% confidence interval [CI] 38.4%-63.2%) and those who met the criteria for probable SARS (58.0%, 95% CI 44.2%-70.7%). SARS-CoV was detected in nasopharyngeal swabs in 33 (32.4%) of 102 patients, in stool specimens in 19 (63.3%) of 30 patients, and in specimens from the lower respiratory tract in 10 (58.8%) of 17 patients. INTERPRETATION These findings suggest that the rapid diagnostic tests in use at the time of the initial outbreak lack sufficient sensitivity to be used clinically to rule out SARS. As tests for SARS-CoV continue to be optimized, evaluation of the clinical presentation and elucidation of a contact history must remain the cornerstone of SARS diagnosis. In patients with SARS, specimens taken from the lower respiratory tract and stool samples test positive by means of RT-PCR more often than do samples taken from other areas.


Applying rapid genome sequencing technologies to characterize pathogen genomes.


title: [Equipment for SARS control in patient care units and in units that process infected samples. 2003].

text: The information on severe acute respiratory syndrome (SARS) that has been gathered up to this point has made it possible to prepare recommendations concerning the equipment needed to protect health workers responsible for patient care and for processing potentially infected samples. Protecting such personnel is a key element in the strategy to control the spread of SARS. The needed equipment includes clothing; footwear; such protective devices as masks, safety glasses, and gloves; disinfectant solutions; laboratory equipment; and materials for obtaining and transporting samples. Prepared by the Western Pacific Regional Office of the World Health Organization, this list will help implement measures to contain the epidemic. The list gives the recommended quantities, specifications for the items, and possible alternatives for some items. This list of equipment is not exhaustive; it is intended to provide a small emergency supply for a period of 3 or 4 days for a single isolation unit with approximately 50 workers. To guarantee the availability of the equipment in the needed quantities, infection control authorities should take into account the number of isolation units needed and the length of time that containment measures will need to be in place. Adequate reserves should be available 24 hours a day, and plans should be made for rapid access to larger stocks in the event of a larger outbreak.


title: Outcomes and Complications of Simultaneous Laparoscopic Cystectomy and Laparoscopic Nephroureterectomy with Umbilical Reduced Port Surgery

text: Objective: In recent years, although reduced port surgeries (RPS) have been reported for many urological diseases, there have been no reports regarding simultaneous laparoscopic cystectomy and unilateral or bilateral nephroureterectomy with umbilical RPS. Therefore, the aim of this study was to evaluate outcomes and complications of simultaneous laparoscopic cystectomy and unilateral or bilateral nephroureterectomy with umbilical RPS. Methods: We performed a preliminary case series of 4 patients with synchronous upper urinary tract (UUT) tumor and invasive bladder cancer who underwent simultaneous laparoscopic cystectomy and unilateral or bilateral nephroureterectomy with umbilical RPS between 2014 and 2017 at our hospital. Demographic data, pathologic features, the surgical technique, and outcomes were retrospectively analyzed. Result: All 4 patients were men whose median age was 79 years (range 65-85 years) and median body mass index was 24.2 kg/m2 (range 21.5-27.3 kg/m2). The laparoscopic approach was technically successful in all 4 patients without the need for open conversion. The median total operative time was 434 minutes (range 372-481 minutes). The median estimated blood loss was 773 ml (range 153-923 ml), median interval to resuming oral intake was 2 days (range 1-7 days), and median hospital stay was 16 days (range 13-20 days). Conclusion: The reduced port approach is technically feasible in terms of many outcome measures, with significant cosmetic advantages. This method can be performed safely and recommended as a viable option for patients with concomitant UUT and bladder cancer.


SARS: prudence, not panic.


title: Isatin compounds as noncovalent SARS coronavirus 3C-like protease inhibitors.

text: A series of isatin derivatives were synthesized and tested against SARS CoV 3C-like protease. Substitutions at the N-1 and C-5 positions were examined to elucidate the differences in substrate binding sites of the rhinovirus 3C protease and SARS CoV 3C-like protease. Compound 5f shows significant inhibition with an IC(50) of 0.37 microM. Further study showed that, unlike the irreversible covalent binding of isatin derivatives to human rhinovirus 3C protease, the compounds tested in this study are all noncovalent reversible inhibitors.


title: Polypeptides and functions of antigens from human coronaviruses 229E and OC43.

text: Coronaviruses possess three major size classes of polypeptides as judged by molecular weight: approximately 180,000, approximately 50,000, and approximately 23,000. Human coronaviruses 229E and OC43 possess not only three similar size classes of polypeptides but also three distinct antigens, none of which cross-react with the heterologous strain. Polypeptides separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were reacted in rocket immunoelectrophoresis with antiserum monospecific to each of the three strain-specific antigens (excised precipitin lines from crossed immunoelectrophoresis profiles were used for immunogens). Monospecific antiserum with neutralizing ability reacted with a polypeptide of 186,000 daltons for 229E and a polypeptide of 190,000 daltons for OC43. The antigen which elicited neutralizing antibody response was located at the surface, associated with the corona of the virion, glycosylated, and bound by concanavalin A. Another less prominent surface antigen was represented by size classes of 23,000 daltons for 229E and 24,000 for OC43. The core antigens of the viruses had molecular weights of 49,000 and 229E and 52,000 and OC43 virus. Thus, the molecular weights and functions of the antigens of human coronaviruses are similar to those of animal coronaviruses. The polypeptides of coronaviruses 229E and OC43 are nearly identical as judged by molecular weight, but the similar polypeptides of the two viruses represent different immunological specificities.


title: Changes in locations of crossover sites over time in de novo generated RNA recombinants.

text: Recombinant RNAs generated in plants 3 weeks postinoculation with turnip crinkle virus (TCV) genomic RNA and an associated satellite RNA, sat-RNA D, have a majority of TCV crossover sites in a 24-nucleotide repeat (motif IIIA/IIIB) that forms part of a stable hairpin (Carpenter et al., 1995, J. Mol.Biol. 245, 608-622). To determine if parameters other than nucleotide sequence in the crossover region affect junction site selection, recombinants were assayed at various times postinoculation of plants and protoplasts. Populations of recombinants became progressively shorter in plants and larger in protoplasts. Levels of inoculated transcript and age of the plant were not substantial factors in the shifts in crossover site locations. The two most commonly cloned recombinant species were not amplified to detectable levels in protoplasts, suggesting that these molecules are not viable templates for replication. These results suggest that recombination between sat-RNA D and TCV is a very frequent event, and populations of recombinants are likely generated de novo in each infected cell and represent the original recombinant molecules rather than progeny of such molecules. Therefore, factors other than simple selection for recombinants that are more fit to replicate are probably responsible for the differences in junction sites in populations of sat-RNA D/TCV recombinants.


Occult respiratory viral infections in coronial autopsies: a pilot project.


title: Risk factors for SARS infection within hospitals in Hanoi, Vietnam.

text: We investigated a nosocomial infection of severe acute respiratory syndrome (SARS) in Vietnam in 2003 and attempted to identify risk factors for SARS infection. Of the 146 subjects who came into contact with SARS patients at Hospital A, 43 (29.5%) developed SARS, and an additional 16 (11%) were asymptomatic but SARS-coronavirus (CoV) seropositive. The asymptomatic infection rate accounted for 15.5% of the total number of infected patients at Hospital A, which was higher than that of 6.5% observed at Hospital B, to where all patients from Hospital A were eventually transported (P<0.05). At Hospital A, the risk for developing SARS was 12.6 times higher in individuals not using a mask than in those using a mask. The SARS epidemic in Vietnam resulted in numerous secondary infections due to its unknown etiology and delayed recognition at the beginning of the epidemic. The consistent and proper use of a mask was shown to be crucial for constant protection against infection with SARS.


Erratum To: Accounting for Behavioral Responses during a Flu Epidemic Using Home Television Viewing.


title: Bactericidal activity of a Ce-promoted Ag/AlPO4 catalyst using molecular oxygen in water.

text: The catalytic inactivation of Escherichia coli in water by a cerium (Ce)-promoted silver-loaded aluminum phosphate (Ag/ AlPO4) catalyst using molecular oxygen was investigated. With optimum Ce content, the Ag(Ce)/AlPO4 catalyst exhibited strong bactericidal activity. The process of decomposition of the cell wall and cell membrane was directly observed by TEM. The different morphological changes of E. coli cells treated with the Ag(Ce)/AlPO4 catalyst and those treated with Ag+ suggested that the Ag+ eluted from the catalyst surface did not play an important role during the bactericidal process. Results of DMPO spin-trapping measurements by electron spin resonance (ESR) indicated the formation of the reactive oxygen species (ROS) *OH and *O2-, which caused the considerable bactericidal activity. The formation of H2O2 acted as an important intermediate; this was confirmed by addition of catalase as the scavenger. A possible catalytic oxidation bactericidal mechanism using molecular oxygen was proposed for the Ag(Ce)/AlPO4 catalyst.


title: Infection prevention and control in blood purification centers during the COVID-19 epidemic: a single institution experience from Zhejiang, China.

text: not available.


Evidence of torovirus infection in diarrhoeic cattle.


title: Hybridoma antibodies to the murine coronavirus JHM: characterization of epitopes on the peplomer protein (E2).

text: A panel of hybridoma antibodies that react with the surface peplomer glycoprotein (E2) of the murine coronavirus JHM were produced to characterize major antigenic domains associated with functions related to virulence. Three groups of hybridoma antibodies were differentiated by immunoprecipitation of lysates from JHM-infected cells. One group precipitated the virion structural proteins gp170 and gp98 together with the intracellular form of E2, gp150. A second group reacted with gp98 and gp150, and a third group precipitated gp150 only. Competition assays with biotinylated hybridoma antibodies allowed the definition of at least six different epitope groups. Only those antibodies which immunoprecipitated both gp170 and gp98 neutralized infectivity, inhibited cell fusion and protected infected rats against acute disease. Another class of antibodies binding to gp170 and gp98 also neutralized JHM virus, but did not inhibit fusion and did not protect against disease. Antibodies that immunoprecipitated gp150 and gp98 revealed only weak neutralization and did not inhibit cell fusion or protect animals. Four epitopes were defined by antibodies that immunoprecipitated gp150, but revealed no biological activity. These data indicate that the site responsible for cell fusion is associated with an epitope group carried by gp170 and gp98. Neutralizing antibodies bind to this and another epitope. Furthermore, protection of JHM-infected rats against acute disease requires both inhibition of cell fusion and neutralization of virus.


title: Biphasic immune responses of cats under controlled infection with a feline enteric coronavirus-79-1683 strain.

text: Kittens inoculated orally with 10(2) PFU of feline enteric coronavirus developed no antibody to the virus despite the repeated challenges. However, they developed antibody for a long period with 5 x 10(3)-1.6 x 10(5) (mean 3 x 10(4)) and with 2.5 x 10(3)-2 x 10(4) (mean 6 x 10(3)) immunoperoxidase antibody titer when they were challenged with 10(5) and 10(3) PFU of virus following previous challenges, respectively. Viremia was found when kittens were inoculated with 10(5) PFU of virus, but not with 10(3) PFU of virus. The dose of 10(3) PFU of virus seemed to be a lower limit to establish infection. These results indicate that local infection induces a low antibody response and systemic infection induces a high antibody response.


title: Refining estimates of public health spending as measured in national health expenditure accounts: the Canadian experience.

text: The recent focus on public health stemming from, among other things, severe acute respiratory syndrome and avian flu has created an imperative to refine health-spending estimates in the Canadian Health Accounts. This article presents the Canadian experience in attempting to address the challenges associated with developing the needed taxonomies for systematically capturing, measuring, and analyzing the national investment in the Canadian public health system. The first phase of this process was completed in 2005, which was a 2-year project to estimate public health spending based on a more classic definition by removing the administration component of the previously combined public health and administration category. Comparing the refined public health estimate with recent data from the Organization for Economic Cooperation and Development still positions Canada with the highest share of total health expenditure devoted to public health than any other country reporting. The article also provides an analysis of the comparability of public health estimates across jurisdictions within Canada as well as a discussion of the recommendations for ongoing improvement of public health spending estimates. The Canadian Institute for Health Information is an independent, not-for-profit organization that provides Canadians with essential statistics and analysis on the performance of the Canadian health system, the delivery of healthcare, and the health status of Canadians. The Canadian Institute for Health Information administers more than 20 databases and registries, including Canada's Health Accounts, which tracks historically 40 categories of health spending by 5 sources of finance for 13 provincial and territorial jurisdictions. Until 2005, expenditure on public health services in the Canadian Health Accounts included measures to prevent the spread of communicable disease, food and drug safety, health inspections, health promotion, community mental health programs, public health nursing, as well as all the costs for the general administration of government health departments.


title: Polytope vaccines for the codelivery of multiple CD8 T-cell epitopes.

text: Vaccines against a number of diseases, including HIV, Epstein Barr virus, malaria and several cancers, are believed to require the coinduction of multiple alphabeta CD8+ cytotoxic T-lymphocyte responses that are directed towards a number of different target antigens. The difficulties associated with making large recombinant vaccines that contain numerous antigens has led to the development of alphabeta CD8+ cytotoxic T-lymphocyte polyepitope or polytope vaccine approach, where multiple (usually 8-10 amino acids long) alphabeta CD8+ cytotoxic T-lymphocyte epitopes, derived from several antigens are conjoined into single artificial constructs. Such polytope constructs can be delivered using a number of different vaccine vector modalities with each epitope in the construct emerging as individually immunogenic.


Rethinking global health training in North America.


title: Two cases of severe obstructive pneumonia associated with an HKU1-like coronavirus.

text: BACKGROUND During the last few years a number of previously undescribed viruses, including human metapneumovirus, coronaviruses SARS, NL63 and HKU1, and bocavirus, were identified in nasopharyngeal samples from patients with signs of respiratory infections. These viruses may cause mild to life-threatening infections. OBJECTIVES Nasopharyngeal samples from hospitalized pediatric patients with respiratory disease were analysed for the presence of coronaviruses and other well known and newly identified respiratory viruses. RESULTS Two clinical cases of a severe obstructive pneumonia, which were associated with the presence of RNA of a novel variant (subtype) of HKU1 coronavirus in the nasopharyngeal aspirates, were identified. DISCUSSION The detection of a HKU1-like coronavirus in pediatric patients in the current study complement the most recent independent finding of similar or closely related coronaviruses in patients with respiratory diseases in France (Vabret et al. 2006) and Norway (Jonassen et al., see accompanying manuscript). These observations indicate a wide dissemination of HKU1-like coronaviruses in Europe.


title: Do we need genomic research for the prevention of common diseases with environmental causes?

text: Concerns have been raised about the value of genomic research for prevention and public health, especially for complex diseases with risk factors that are amenable to environmental modification. Given that gene-environment interactions underlie almost all human diseases, the public health significance of genomic research on common diseases with modifiable environmental risks is based not necessarily on finding new genetic "causes" but on improving existing approaches to identifying and modifying environmental risk factors to better prevent and treat disease. Such applied genomic research for environmentally caused diseases is important, because 1) it could help stratify disease risks and differentiate interventions for achieving population health benefits; 2) it could help identify new environmental risk factors for disease or help confirm suspected environmental risk factors; and 3) it could aid our understanding of disease occurrence in terms of transmission, natural history, severity, etiologic heterogeneity, and targets for intervention at the population level. While genomics is still in its infancy, opportunities exist for developing, testing, and applying the tools of genomics to clinical and public health research, especially for conditions with known or suspected environmental causes. This research is likely to lead to population-wide health promotion and disease prevention efforts, not only to interventions targeted according to genetic susceptibility.


title: In vivo RNA-RNA recombination of coronavirus in mouse brain.

text: RNA-RNA recombination between different strains of the murine coronavirus mouse hepatitis virus (MHV) occurs at a very high frequency in tissue culture. To demonstrate that RNA recombination may play a role in the evolution and pathogenesis of coronaviruses, we sought to determine whether MHV recombination could occur during replication in the animal host of the virus. By using two selectable markers, i.e., temperature sensitivity and monoclonal antibody neutralization, we isolated several recombinant viruses from the brains of mice infected with two different strains of MHV. The recombination frequency was very high, and recombination occurred at multiple sites on the viral RNA genome. This finding suggests that RNA-RNA recombination may play a significant role in natural evolution and neuropathogenesis of coronaviruses.


Emerging and new infectious diseases.


title: Viral-induced rhinitis.

text: Upper respiratory viruses cause self-limited illness characterized by acute rhinitis. In rhinovirus colds the symptoms are thought to be caused by the host response rather than viral damage of the nasal epithelium. Rhinovirus triggers an inflammatory cascade, evidenced by the presence of inflammatory mediators (e.g., IL-8) and proinflammatory cytokines (e.g., kinins) in nasal secretions, which results in symptomatic illness. In contrast to rhinovirus and coronavirus, which do not cause discernible epithelial damage, influenza virus and adenovirus do damage the nasal epithelium. Appropriate antiviral therapy will depend on the causative virus. Treatment of rhinovirus colds may require an antiviral agent (e.g., interferon alpha) in combination with antiinflammatory medication.


title: Plasma vitamin C and tocotrienols changes in response to dietary supplementation among young male adults.

text: BACKGROUND AND AIMS Intake of the antioxidant vitamins C and E lowers the oxidative stress. The study aimed to determine plasma concentrations of vitamin C and tocotrienols after supplementation of both vitamins in young male adults. MATERIALS AND METHODS A total of 64 police recruits were randomly assigned to one of these groups: (a) 500 mg vitamin C (Vitamin C), (b) 200 mg Tocovid (Tocotrienol), (c) combination of 500 mg vitamin C and 200 mg Tocovid (Combination) or (d) placebo (Placebo) for eight-weeks of supplementation followed by six-week washout period. RESULTS In Combination group, mean plasma vitamin C concentration significantly increased from baseline 2.86 +/- 1.19 mg/L to 10.37 +/- 1.29 mg/L and 15.63 +/- 1.27 mg/L after four- and eight-week supplementation, respectively. The corresponding figures for alpha-, delta- and gamma-tocotrienols were 9.9 +/- 2.5 ng/ml to 104.1 +/- 19.8 ng/ml and 112.8 +/- 38.0 ng/ml; 2.5 +/- 0.9 ng/ml to 29.9 +/- 7.0 ng/ml and 17.9 +/- 4.7 ng/ml; 19.2 +/- 3.1 ng/ml to 75.2 +/- 24.1 ng/ml and 161.7 +/- 49.9 ng/ml, respectively. In Vitamin C group, plasma vitamin C concentrations were significantly increased. Conversely, concentration of plasma vitamin C in Tocotrienol group increased from baseline of 2.72 +/- 0.20 mg/L to 6.80 +/- 0.63 mg/L and 8.9 +/- 0.77 mg/L respectively. Plasma concentrations of alpha-, delta- and gamma-tocotrienols in this group were significantly elevated. After 6-week washout period, all the elevated concentrations returned to basal levels. CONCLUSION The study showed a good bioavailability of these vitamins and increment due to supplementation.