title: Single amino acid changes in the viral glycoprotein M affect induction of alpha interferon by the coronavirus transmissible gastroenteritis virus.

text: Transmissible gastroenteritis virus, an enteropathogenic coronavirus of swine, is a potent inducer of alpha interferon (IFN-alpha) both in vitro and in vivo. Previous studies have shown that virus-infected fixed cells or viral suspensions were able to induce an early and strong IFN-alpha synthesis by naive lymphocytes. Two monoclonal antibodies directed against the viral membrane glycoprotein M (29,000; formerly E1) were found to markedly inhibit virus-induced IFN production, thus assigning to M protein a potential effector role in this phenomenon (B. Charley and H. Laude, J. Virol. 62:8-11, 1988). The present report describes the selection and characterization of a collection of 125 mutant viruses which escaped complement-mediated neutralization by two IFN induction-blocking anti-M protein monoclonal antibodies. Two of these mutants, designated H92 and dm49-4, were found to exhibit a markedly reduced interferogenic activity. IFN synthesis by lymphocytes incubated with purified suspensions of these mutants was 30- to 300-fold lower than that of the parental virus. The transcription of IFN-alpha genes following induction by each mutant was decreased proportionally, as evidenced by Northern (RNA) blot analysis. The sequence of the M gene of 20 complement-mediated neutralization-resistant mutants, including the 2 defective mutants, was determined by direct sequencing of genome RNA. Thirteen distinct amino acid changes were predicted, all located at positions 6 to 22 from the N terminus of the mature M protein and within the putative ectodomain of the molecule. Two substitutions, Thr-17 to Ile and Ser-19 to Pro, were assumed to generate the defective phenotypes of mutants dm49-4 and H92, respectively. The alteration of an Asn-Ser-Thr sequence in dm49-4 virus led to the synthesis of an M protein devoid of a glycan side chain, which suggests a possible involvement of this structure in IFN induction. Overall, these data supported the view that an interferogenic determinant resides in the N-terminal, exposed part of the molecule and provided further evidence for the direct role of M protein in the induction of IFN-alpha by transmissible gastroenteritis virus. The acronym VIP (viral interferogenic protein) is proposed as a designation for this particular class of proteins.


title: [Experts recommendation on the diagnosis and treatment of oral mucosal diseases during prevention and control stage of the novel coronavirus infection].

text: The announcement of National Health Commission on January 20, 2020 (No. 1 of 2020) has included Novel Coronavirus Pneumonia(NCP) into the B class infectious diseases according to the law of the People's Republic of China on the prevention and control of infectious diseases, and has been managed as A class infectious diseases. People's governments at all levels and health administration departments have been paying high attention to it, and medical and health institutions have taken a series of prevention and control measures according to the law to prevent and control the spread of NCP. In the process of combating the outbreak of NCP, how to protect the oral mucosal disease physicians normatively, how to ensure hand and environmental cleaning and disinfection; how to carry out triage of the patients with oral mucosal disease accompanied by fever reasonably; how to carry out oral mucosal disease patients'self-care and self-management during this stage, are the practical problems that all levels of medical institutions and medical workers and patients will meet. For these three problems, we invited the experts from the Society of Oral Medicine, Shanghai Stomatological Association and the Oral Mucosal Disease Specialist Alliance Organization of Shanghai Ninth People's Hospital Group to write the recommendation on the diagnosis and treatment of oral mucosal disease during prevention and control stage of novel coronavirus infection, in order to provide references for oral mucosal disease medical physicians and patients in this special period. This recommendation will be updated according to the situation of epidemic prevention and control in China and the new relevant diagnosis and treatment plans.


title: Contributing to communicable diseases intelligence management in Canada: CACMID meeting, March 2007, Halifax, Nova Scotia.

text: In the spring of 2003, the Public Health Agency of Canada (then, Health Canada) partnered with several provincial/territorial and regional public health stakeholders to improve pan-Canadian public health surveillance, communications and response through the application of new technologies. This resulted in the creation of the Canadian Network for Public Health Intelligence (CNPHI), a comprehensive framework of applications and resources designed to fill critical gaps in Canada's national public health infostructure. Over the past four years, the CNPHI has evolved into Canada's only pan-Canadian public health information management system. With over 2000 registered users, the current CNPHI environment consists of more than 30 integrated applications and systems that can be loosely categorized into four functional groups: data exchange; data analysis and integration; communication, collaboration and coordination; and knowledge management. Despite poor data repositories, legacy information management systems, and the lack of standards and agreements, the CNPHI has demonstrated that much can be accomplished in these areas. Over the next decade, significant barriers impeding additional advances will be bridged through the implementation of the Electronic Health Record, and through ongoing efforts to address gaps in standards, and data- and information-sharing agreements. Together with new technologies coming on-line, opportunities to further enhance public health surveillance and response will be limited only by one's imagination.


title: Generalized Tetanus in a Gyrfalcon ( Falco rusticolus ) with Pododermatitis.

text: A 2-yr-old male gyrfalcon ( Falco rusticolus ) was presented for severe and generalized muscle spasticity and pododermatitis. The falcon had been treated for pododermatitis over the previous 4 mo. Muscle rigidity and spasms involved the entire bird but were more severe on the right leg. The bird was also tachypneic and hyperthermic at 45 C. While the plantar pododermatitis lesions had healed, there was still a small abscess on the lateral aspect of the right foot. Clinical signs were consistent with tetanus. Several bacteria were isolated from the abscess including Clostridium tetani . The isolate was confirmed to be toxigenic by PCR. Attempts to detect tetanus toxin in the bird's plasma were unsuccessful. The abscess was debrided. The gyrfalcon received equine tetanus antitoxin, intravenous metronidazole, methocarbamol, midazolam, a constant-rate infusion of Fentanyl, active cooling, and supportive care. Inhalant anesthesia with isoflurane was the only treatment that would lower the body temperature and reduce the clinical signs. The gyrfalcon died a few hours after admission. The characteristic clinical signs and isolation of toxigenic C. tetani from a wound were strong supportive evidence for a diagnosis of tetanus. This case constitutes the first reported natural occurrence of tetanus in an avian species. Further information is needed to determine whether gyrfalcons are more susceptible to tetanus than are other avian species and whether pododermatitis lesions may be risk factors.


title: Effect of the anatomic reservoir on low-flow oxygen delivery via nasal cannula: constant flow versus pulse flow with portable oxygen concentrator.

text: BACKGROUND The F(IO(2)) for a nasal cannula with constant flow (CF) depends on the anatomic reservoir (AR), which is affected by changes in frequency and end-expiratory flow. Conversely, pulse flow (PF) devices do not require the AR. The purpose of this study was to compare the F(IO(2)) delivered by a nasal cannula supplied by CF via oxygen tank with that delivered by PF delivered via portable oxygen concentrator. Hypotheses were (1) a lung model of COPD with non-zero end-expiratory flow decreases F(IO(2)) for CF more than for PF, and (2) CF and PF perform differently in terms of F(IO(2)) delivery, despite having equivalent settings. METHODS Normal and COPD lung models were simulated (IngMar Medical ASL 5000) using published human data: normal: breathing frequency = 15 breaths/min, R(in) = 4 cm H2O · s · L(-1), R(out) = 4 cm H2O · s · L(-1), C = 60 mL · cm H2O(-1), tidal volume (VT) = 685 mL, P(max) = 11.95 cm H2O, increase = 33%, and release = 28; COPD: breathing frequency = 20 breaths/min, R(in) = 12 cm H2O · s · L(-1), R(out) = 25 cm H2O · s · L(-1), C = 66 mL · cm H2O(-1), VT = 685 mL, Pmax = 24.52 cm H2O, increase = 35%, and release = 23%. CF was 1-5 L/min. Portable oxygen concentrators used were Solo2 (Invacare), XPO2 (Invacare), FreeStyle (AirSep), Focus (AirSep), One G3 (Inogen), and LifeChoice ActivOx (Inova Labs). RESULTS CF produced significantly higher F(IO(2)) at all settings for normal lungs but lower for COPD lungs compared with Solo2. COPD reduced the F(IO(2)) for CF but had a smaller variable effect for PF. Data show there is no equivalency between PF setting and CF rates for the portable oxygen concentrators tested. CONCLUSIONS CF oxygen delivery via a nasal cannula is significantly reduced by elimination of the AR in a model of COPD, yielding clinically important decreases in F(IO(2)). PF (delivered with a portable oxygen concentrator) is relatively unaffected. This study supports the recommendation that clinicians and caretakers should titrate the PF setting to each patient's unique oxygen requirements.


docid: hljklens

title: [Functional urology during COVID-19: Recommendations during de-escalation.]

text: OBJECTIVES Due to the COVID-19 Pandemic, all surgical activity that was not life threatening was cancelled , as well as most face-to-face consultations. Currently the beginning of the de-escalation phases that will led us to a new normal, forces us to establish some degree of priority in the interventions as well as in the medical consultations. Our objective is to establish some recommendation on Functional Urology office visits and surgical interventions that serve as a tool to facilitate decision-making. MATERIAL AND METHODS: Experts in Functional Urology from different autonomous communities of Spain were contacted to design a strategy to reorganize the activity of both, diagnosis and treatment. A modified nominal group technique has been used due to the extraordinary restrictions of assembly and mobility during the COVID pandemic. The first signer (EMC) made the first draft with the measures adopted and the strategy to be followed during the evolution of the COVID-19 pandemic. The proposal was sent to the rest of the authors, in order to unify criteria and experiences to reach a quick consensus on the relative priority of the different activities, problems and solutions. A final version was approved by all authors May 27, 2020. RESULTS: Tables of recommendation have been prepared for outpatient consultation, surgical and technical interventions, according to de-escalation phases proposed by the Spanish Associations of Surgeons. CONCLUSIONS: The change that COVID-19 Pandemich as involved in our clinical practice force us to seek alternative methods to treat our patients, some of which may already be established. Mean while, a consensusin decision making is necessary. Documents such as the current one, are intended to guide the management of patients with urological functional pathology in exceptional situations. Logically, it should be adapted to material and human availability, and to the idiosyncrasy of each Urology service.


title: Functional links between the fusion peptide-proximal polar segment and membrane-proximal region of human immunodeficiency virus gp41 in distinct phases of membrane fusion.

text: The binding of CD4 and chemokine receptors to the gp120 attachment glycoprotein of human immunodeficiency virus triggers refolding of the associated gp41 fusion glycoprotein into a trimer of hairpins with a 6-helix bundle (6HB) core. These events lead to membrane fusion and viral entry. Here, we examined the functions of the fusion peptide-proximal polar segment and membrane-proximal Trp-rich region (MPR), which are exterior to the 6HB. Alanine substitution of Trp(666), Trp(672), Phe(673), and Ile(675) in the MPR reduced entry by up to 120-fold without affecting gp120-gp41 association or cell-cell fusion. The L537A polar segment mutation led to the loss of gp120 from the gp120-gp41 complex, reduced entry by approximately 10-fold, but did not affect cell-cell fusion. Simultaneous Ala substitution of Leu(537) with Trp(666), Trp(672), Phe(673), or Ile(675) abolished entry with 50-80% reductions in cell-cell fusion. gp120-gp41 complexes of fusion-defective double mutants were resistant to soluble CD4-induced shedding of gp120, suggesting that their ability to undergo receptor-induced conformational changes was compromised. Consistent with this idea, a representative mutation, L537A/W666A, led to an approximately 80% reduction in lipophilic fluorescent dye transfer between gp120-gp41-expressing cells and receptor-expressing targets, indicating a block prior to the lipid-mixing phase. The L537A/W666A double mutation increased the chymotrypsin sensitivity of the polar segment in a trimer of hairpins model, comprising the 6HB core, the polar segment, and MPR linked N-terminally to maltose-binding protein. The data indicate that the polar segment and MPR of gp41 act synergistically in forming a fusion-competent gp120-gp41 complex and in stabilizing the membrane-interactive end of the trimer of hairpins.


[The influenza pandemic].


title: Engineering responses to pandemics.

text: Focusing on pandemic influenza, this chapter approaches the planning for and response to such a major worldwide health event as a complex engineering systems problem. Action-oriented analysis of pandemics requires a broad inclusion of academic disciplines since no one domain can cover a significant fraction of the problem. Numerous research papers and action plans have treated pandemics as purely medical happenings, focusing on hospitals, health care professionals, creation and distribution of vaccines and anti-virals, etc. But human behavior with regard to hygiene and social distancing constitutes a first-order partial brake or control of the spread and intensity of infection. Such behavioral options are "non-pharmaceutical interventions." (NPIs) The chapter employs simple mathematical models to study alternative controls of infection, addressing a well-known parameter in epidemiology, R0, the "reproductive number," defined as the mean number of new infections generated by an index case. Values of R0 greater than 1.0 usually indicate that the infection begins with exponential growth, the generation-to-generation growth rate being R0. R0 is broken down into constituent parts related to the frequency and intensity of human contacts, both partially under our control. It is suggested that any numerical value for R0 has little meaning outside the social context to which it pertains. Difference equation models are then employed to study the effects of heterogeneity of population social contact rates, the analysis showing that the disease tends to be driven by high frequency individuals. Related analyses show the futility of trying geographically to isolate the disease. Finally, the models are operated under a variety of assumptions related to social distancing and changes in hygienic behavior. The results are promising in terms of potentially reducing the total impact of the pandemic.


The Korean Society for Biochemistry and Molecular Biology.


title: Effect of enzymatic degradation on the mechanical properties of biological scaffold materials.

text: BACKGROUND Biological scaffolds must support a complex balance of resisting enzymatic degradation while promoting tissue remodeling. Thus, the purpose of this study was to evaluate the effects of in vitro enzymatic exposure on the mechanical properties of biological scaffolds. It was hypothesized that exposure to an enzyme solution would result in decreased tensile strength and that crosslinked scaffolds would resist enzymatic degradation more effectively than noncrosslinked scaffolds. METHODS Nine scaffolds were evaluated (four porcine dermis: Permacol™, CollaMend™, Strattice™, XenMatrix™; two human dermis: AlloMax™, FlexHD(®); two bovine pericardium: Veritas(®), PeriGuard(®); and one porcine small intestine submucosa: Surgisis™). Ten specimens (n = 10) were hydrated in saline at 37 °C and subjected to uniaxial testing to establish baseline properties. 50 specimens (n = 50) were incubated in collagenase solution at 37 °C for 2, 6, 12, 24, or 30 h (n = 10 each group) followed by uniaxial tensile testing. RESULTS Tensile strength was significantly reduced after 30 h for CollaMend™, AlloMax™, Veritas(®), Strattice™, XenMatrix™, Permacol™, and FlexHD(®) (p < 0.01), while PeriGuard(®) demonstrated a slight increase in tensile strength (p = 0.0188). Crosslinked bovine pericardium (PeriGuard(®)) maintained greater tensile strength than noncrosslinked bovine pericardium (Veritas(®)) throughout all exposure periods (p < 0.0001). Similarly, crosslinked porcine dermis (Permacol™) maintained greater tensile strength than noncrosslinked porcine dermis (Strattice™ and XenMatrix™) throughout all exposure periods (p < 0.0001). CONCLUSIONS Materials that deteriorate rapidly after in vitro enzymatic exposure may also deteriorate rapidly in vivo, particularly when exposed to a wound environment with elevated levels of matrix metalloproteinases. Permacol™, CollaMend™, Strattice™, FlexHD(®), and PeriGuard(®) survived the longest incubation period (30 h) and withstood mechanical testing. XenMatrix™, AlloMax™, Veritas(®), and Surgisis™ degraded more quickly and did not survive the longer exposure periods. Scaffolds that maintain strength characteristics after in vitro collagenase exposure may be advantageous for long-term hernia repair scenarios where elevated enzyme levels are expected.


title: Viruses and virus-like particles identified in ostrich gut contents.

text: Samples of either gut content from ostriches showing symptoms of enteritis, or allantoic fluid of eggs inoculated with ostrich isolates, were examined for the presence of viral agents by direct negative-contrast electron microscopy. Only a few virus types could be identified with certainty, namely a circovirus (1 sample), a coronavirus (1 sample), a member of either the toga- or bunyaviridae (1 sample), enterovirus (16 samples) and paramyxovirus (26 samples). A large number of samples contained structures resembling myxovirus particles that were interpreted as fringed membranous particles of non-viral origin. An unusual observation of probable single-strand nucleocapsid helices, possibly originating from digested plant material and which were identified in a number of small intestine samples, is reported. This is the 1st report of a spectrum of viruses and virus-like particles occurring in enteric samples from ostriches in South Africa. The low incidence and variety of viruses reported here contribute to the multifactorial origin and complexity of enteric disease in ostriches as well as in other birds and mammalian species.


Preparedness for biological terrorism in the United States: Project BioShield and Beyond.


Learning from mistakes.


title: Cost analysis of bariatric surgery in Denmark made with a decision-analytic model.

text: INTRODUCTION Bariatric surgery offers effective obesity treatment. The aim of this study was to evaluate the cost-effectiveness of bariatric surgery in Denmark from a third-party payer perspective in the mid- (ten years) and long-term (lifetime). METHODS A state-transition Markov model was developed in which patients may experience surgery, post-surgery complications, diabetes mellitus type 2, cardiovascular diseases or die. Transition probabilities, costs and utilities were informed by the literature. Three types of surgery were included: gastric bypass, sleeve gastrectomy and adjustable gastric banding. The impact of different surgical methods on BMI level was informed by the Danish Obesity Surgery Registry (Dansk Fedmekirurgiregister). RESULTS In the ten-year base-case analysis, bariatric surgery led to a cost increment of 19,332 DKK and generated an additional 1.1 quality-adjusted life years (QALYs). In the course of a lifetime, surgery leads to savings of 36,403 DKK, an additional 0.7 life years and 2.9 QALYs. Bariatric surgery was cost-effective at ten years with an incremental cost-effectiveness ratio of 17,818 DKK per QALY and was dominant over conservative management in the course of a lifetime. Up to three years of delay in the provision of surgery resulted in a reduction of life years, a lower QALY gain and a minor decrease in healthcare costs. CONCLUSIONS In Denmark, bariatric surgery is cost-effective at ten years and may produce a significant reduction in healthcare costs over the course of a lifetime in persons with severe obesity. FUNDING Synergus AB received support for economic model development from Covidien AG (now part of Medtronic). TRIAL REGISTRATION not relevant.


A survey on bovine rotavirus type 1-associated neonatal calf diarrhea in a beef herd.


title: [Ultrastructural observation of rat thymus tissue with coronavirus infection].

text: OBJECTIVE To investigate the ultrastructure of rat thymus tissues with rat coronavirus (RCV) infection for clarifying the mechanism responsible for the morphological changes of the cells infected by RCV. METHODS Routine electron microscopy was performed for observing RCV-infected rat thymus tissues. RESULTS Following RCV infection, endoplasmic reticulum (ER) pools of different dimensions were observed in the cytoplasm of the thymic epithelial reticular cells, merging subsequently with each other into larger ER lakes filled with particles of mature RCV, or viral inclusion bodies. After germination on the ER membrane, the viruses entered the matrix of the ER lake to mature and were eventually excreted to the extracellular space. The RCV particles were spherical in shape with a diameter of 100-130 nm and two distinct membranes, the outer one being the envelope and the inner one the nuclear capsid to enclose the viroplasm. Between the envelop and nuclear capsid was a electron-lucent middle layer comprising one to two thin membranous structures. Large quantity of short spike-like projections starting from the nucleus capsid penetrated the middle layer and the envelop to reach the glycoprotein coat and formed a corona-like structure. Mature RCV particles were distributed around the ER pools, cytoplasm, and intercellular space, and the RCVs in the endosome/lysosome were devoid of the envelop and nuclear capsid. CONCLUSION The ER lakes are involved in the maturation of the viruses, and the envelop and nuclear capsid of the virus entering the cells from extracellular space are removed and degraded in the endosome/lysosome. Replications of virus occurs in plasma of the thymic epithelial reticular cells, and no RCV can be detected in the thymocytes.


title: cis Requirement for N-specific protein sequence in bovine coronavirus defective interfering RNA replication.

text: A naturally occurring 2.2-kb defective interfering (DI) RNA of the bovine coronavirus, structurally a simple fusion of the genomic termini, contains a single contiguous open reading frame (ORF) or 1.7 kb composed of the 5'-terminal 288 nucleotides of polymerase gene 1a and all 1,344 nucleotides of the nucleocapsid protein (N) gene. The ORF must remain open throughout most of its sequence for replication to occur. To determine the qualitative importance of the N portion of the chimeric ORF in DI RNA replication, transcripts of mutated reporter-containing constructs were tested for replication in helper virus-infected cells. It was determined that the N ORF could not be replaced by the naturally occurring internal I protein ORF, accomplished by deleting the first base in the N start codon which leads to a +1 frameshift, nor could it be replaced by the chloramphenicol acetyltransferase ORF. Furthermore, 3'-terminal truncations of the N gene leaving less than 85% of its total length were likewise not tolerated. Small in-frame deletions and in-frame foreign sequence insertions of up to 99 nucleotides within certain regions of the N ORF were tolerated, however, but the rate of DI RNA accumulation in these cases was lower. These results indicate that there is a requirement for translation of most if not all of the N protein in cis for optimal replication of the bovine coronavirus DI RNA and suggest that a similar requirement may exist for viral genome replication.


title: Relationship among Dexamethasone Suppression Test, personality disorders and stressful life events in clinical subtypes of major depression: An exploratory study.

text: : BACKGROUND: The present study aimed to investigate the relationship between dexamethasone suppression test, personality disorder, stressful life events and depression. MATERIAL: Fifty patients (15 males and 35 females) aged 41.0 +/- 11.4 years, suffering from Major Depression according to DSM-IV criteria entered the study. METHOD: Diagnosis was obtained with the aid of the SCAN v 2.0 and the IPDE. Psychometric assessment included the HDRS, HAS, the Newcastle Scale (version 1965 and 1971), the Diagnostic Melancholia Scale, the Personality Deviance Scale and the GAF scale. The 1 mg DST was used. STATISTICAL ANALYSIS: Included MANOVA, ANOVA with LSD post hoc test and chi-square test. RESULTS: Sixteen (32%) patients were non-suppressors. Eight patients without Personality Disorder (PD) (23.5%), and 5 of those with PD of cluster B (50%) were non-suppressors. Atypical patients were the subtype with the highest rate of non-suppression (42.85%). No difference between suppressors and non-suppressors was detected in any of the scales. DISCUSSION: The results of the current study suggest that pathological DST is not a core feature of major depression. They also suggest that there are more than one subtypes of depression, concerning the response to stress. It seems that the majority of depressed patients (50%) does not experience high levels of stress either in terms of self reported experience or neuroendocrine function. The rest of patients however, either experience high levels of stress, or manifest its somatic analogue (DST non-suppression) or have a very low threshold of stress tolerance, which makes them to behave in a hostile way.


title: Isolation and identification of hepatitis E virus in Xinjiang, China.

text: This paper describes isolation and identification of a virus (termed strain 87A) which has the cytopathic effect and haemagglutination properties of hepatitis E virus (HEV). This virus was isolated by tissue culture from the faeces of a patient with acute non-A, non-B enteric hepatitis in Xinjiang, China. The isolated virus was neutralized by acute phase sera obtained from other patients with acute non-A, non-B enteric hepatitis. The virus particles also could be specifically aggregated with acute phase sera from patients with known HEV hepatitis in China, Burma, India and the U.S.S.R., and with acute and convalescent sera from an HEV-infected chimpanzee. Crystalline arrangements of virus particles in the cytoplasm were observed by electron microscopy in ultrathin sections of infected cells. The sedimentation coefficient of the strain 87A virus particles in sucrose gradients was 176S. Purified virus particles revealed a protein band of about 76K on SDS-PAGE and Western blotting. The evidence indicates that the strain 87A virus is an HEV. Our ability to propagate HEV in cell culture should facilitate research on this hepatotropic virus.


title: Documentation issues for mechanical ventilation in pressure-control modes.

text: As hospitals begin to implement electronic medical records, the inadequacies of legacy paper charting systems will become more evident. One area of particular concern for respiratory therapists is the charting of mechanical ventilator settings. Our profession's lack of a standardized and generally accepted taxonomy for mechanical ventilation leaves us with a confusing array of terms related to ventilator settings. Such confusion makes database design impossible for information technology professionals and is a risk-management concern for clinicians. Of particular note is the complexity related to set airway pressures when using modes whose primary control variable is pressure (versus volume). We review the clinically relevant issues surrounding documentation of the patient-ventilator interactions related to airway pressure and provide suggestions for a standardized vocabulary.


1651: INTRAVASCULAR COOLING CATHETER-RELATED VENOUS THROMBOEMBOLISM AFTER HYPOTHERMIA PROTOCOL.


title: Molecular dipstick test for diagnosis of sleeping sickness.

text: Human African trypanosomiasis (HAT) or sleeping sickness is a neglected disease that affects poor rural populations across sub-Saharan Africa. Confirmation of diagnosis is based on detection of parasites in either blood or lymph by microscopy. Here we present the development and the first-phase evaluation of a simple and rapid test (HAT-PCR-OC [human African trypanosomiasis-PCR-oligochromatography]) for detection of amplified Trypanosoma brucei DNA. PCR products are visualized on a dipstick through hybridization with a gold-conjugated probe (oligochromatography). Visualization is straightforward and takes only 5 min. Controls both for the PCR and for DNA migration are incorporated into the assay. The lower detection limit of the test is 5 fg of pure T. brucei DNA. One parasite in 180 microl of blood is still detectable. Sensitivity and specificity for T. brucei were calculated at 100% when tested on blood samples from 26 confirmed sleeping sickness patients, 18 negative controls (nonendemic region), and 50 negative control blood samples from an endemic region. HAT-PCR-OC is a promising new tool for diagnosis of sleeping sickness in laboratory settings, and the diagnostic format described here may have wider application for other infectious diseases.


docid: xhnqywqb

title: T cell responses to whole SARS coronavirus in humans.

text: Effective vaccines should confer long-term protection against future outbreaks of severe acute respiratory syndrome (SARS) caused by a novel zoonotic coronavirus (SARS-CoV) with unknown animal reservoirs. We conducted a cohort study examining multiple parameters of immune responses to SARS-CoV infection, aiming to identify the immune correlates of protection. We used a matrix of overlapping peptides spanning whole SARS-CoV proteome to determine T cell responses from 128 SARS convalescent samples by ex vivo IFN-gamma ELISPOT assays. Approximately 50% of convalescent SARS patients were positive for T cell responses, and 90% possessed strongly neutralizing Abs. Fifty-five novel T cell epitopes were identified, with spike protein dominating total T cell responses. CD8(+) T cell responses were more frequent and of a greater magnitude than CD4(+) T cell responses (p < 0.001). Polychromatic cytometry analysis indicated that the virus-specific T cells from the severe group tended to be a central memory phenotype (CD27(+)/CD45RO(+)) with a significantly higher frequency of polyfunctional CD4(+) T cells producing IFN-gamma, TNF-alpha, and IL-2, and CD8(+) T cells producing IFN-gamma, TNF-alpha, and CD107a (degranulation), as compared with the mild-moderate group. Strong T cell responses correlated significantly (p < 0.05) with higher neutralizing Ab. The serum cytokine profile during acute infection indicated a significant elevation of innate immune responses. Increased Th2 cytokines were observed in patients with fatal infection. Our study provides a roadmap for the immunogenicity of SARS-CoV and types of immune responses that may be responsible for the virus clearance, and should serve as a benchmark for SARS-CoV vaccine design and evaluation.


title: The causes of glaucoma in cats.

text: The cause of glaucoma in 131 enucleated eyes from 128 cats was determined in a retrospective histologic study. Obliteration of the ciliary cleft by diffuse iridal melanoma (38 eyes), or other neoplasms (14 eyes), or by the presence of idiopathic lymphocytic-plasmacytic anterior uveitis (53 eyes) were the most frequent lesions likely to explain the development of glaucoma. Secondary changes of inner retinal atrophy, optic disc cupping, scleral thinning with megaglobus, and atrophy of ciliary processes were similar to those described in dogs and human beings with chronic glaucoma. In light of the duration and severity of the glaucoma, the degree of inner retinal atrophy was often less than expected. Diffuse corneal edema and breaks in Descemet's membrane, changes typical of glaucoma in other species, were rarely detected. Eyes with chronic uveitis and glaucoma had collapsed ciliary clefts, iridoscleral adhesions, and posterior displacement of the iris. We were unable to determine whether these changes were consequences of the uveitis and thus responsible for the development of glaucoma, or if they were merely the result of the chronic glaucoma itself.


New European research project to respond to unexpected epidemic threats such as Escherichia coli.


title: Development of vaccines against common colds.

text: Respiratory tract viruses are particularly significant causes of illness and death in children and in the elderly. Vaccines offer the possibility of decreasing the severity and complications of viral respiratory disease, but development has been delayed by numerous factors. First, there are more than 200 serologically distinct RNA and DNA virus species and strains which cause an essentially similar spectrum of disease. Some re-infect at high efficiency despite little antigenic variation, while others exhibit extensive coat protein variability. Vaccine candidates show variable efficacy in partially immune adults, the immunocompromised and the elderly, and may be ineffective or pathogenic in neonates or in the presence of maternal antibodies. However, effective childhood vaccines are essential to prevent severe disease due to respiratory syncytial virus (RSV) and parainfluenza and to reduce virus transmission to adults. A number of promising vaccines are in clinical trial, and it is likely that vaccines against RSV and parainfluenza will be licensed within the next 5-10 years. Mucosal delivery and the use of novel adjuvants offers the prospect of better vaccines against influenza. The ultimate goal is to develop multivalent mucosal vaccines offering protection against a spectrum of respiratory infections.


title: [Laboratory on a microfluidic chip].

text: The recent achievements of microfluidic chip and its applications, based on the works mainly carried out in the authors' lab are reviewed. The chip fabrication capabilities have been extended into design and fabricate chips with higher degree of complexity in different materials, such as quartz, glass, polymethyl methacrylate (PMMA), and polydimethyl siloxane (PDMS). A set of methods for surface modification of micro-channels on such materials have been developed, which results in better reproducibility and higher efficiency in protein and peptide analysis. The use of novel materials for chip fabrication is also under investigation. A series of microfluidic workstations with integrated chip manipulation as well as laser induced fluorescence (LIF), ultraviolet (UV), electrochemical and chemiluminescence detection modules have been developed to attain the abilities of complex microfluidic control and data acquisition schemes. A single cell/single molecule imagining system was built up for dynamic analysis of molecular or cellular events too. Based on the work mentioned above, different functional units, such as membrane, monolithic, isotachophoresis (ITP) etc were set up and integrated. Glycoform separation of turkey ovalbumin in a lectin monolithic column and an electrophoresis channel was performed on an integrated microchip. And a novel technique has been developed that allows for the coupling of ITP and non-gel sieving electrophoresis for protein analysis in a single microchip and resulting in - 50 fold increase of the sensitivity in comparison with the use of gel electrophoresis only. A single molecule detection (SMD) based technique was developed for simultaneously measuring both bulk flow and near-wall flow velocity in the microchannels. And more recently, an SMD based technology was developed for observing molecular interactions at single molecule level. An ultra-rapid microchip electrophoresis method was established for simultaneous determination intracellular reactive oxygen species (ROS) and reduced glutathione (GSH) related to apoptosis and oxidative stress. In an effort to develop a novel microfluidic based drug screening platform, systematic studies on the interaction between granulocyte colony-stimulating factor (G-CSF) and sulfated oligosaccharides were carried out at both molecular and cellular levels. Doxorubicin induced apoptosis of human hepatocellular carcinoma (HepG2) was studied using the integrated microfluidic device with concentration generator. In the application phase, severe acute respiratory syndrome (SARS) diagnosis based on reverse transcription-polymerase chain reaction (RT-PCR) and microfluidic chip electrophoresis (MCE) with 18 cases, methylation analysis of the P16 gene in 159 samples of patients and references for cancer diagnosis and polymorphism analysis of angiotenigen gene in 226 patients and references with essential hypertension are described. Forty-three up to date references are


title: Risk-stratified seroprevalence of severe acute respiratory syndrome coronavirus among children in Hong Kong.

text: BACKGROUND Severe acute respiratory syndrome was relatively mild in children, and the incidence was significantly lower when compared with adults. Although previous seroepidemiological studies demonstrated that asymptomatic infection was uncommon among health care workers and adult contacts of patients with severe acute respiratory syndrome, it is unclear whether this would extend to the pediatric population. OBJECTIVE The purpose of this study was to determine the seroprevalence of severe acute respiratory syndrome coronavirus among asymptomatic children living near Amoy Gardens (site of largest community outbreak of severe acute respiratory syndrome in Hong Kong) compared with a low-risk region where no community transmission occurred. METHODS The study was conducted from September to October 2003. Target subjects living in the defined high-risk and low-risk areas were approached through the schools within the respective localities. We recruited 353 and 361 children, respectively, from the high-risk and low-risk areas and collected 3 to 5 mL of blood for severe acute respiratory syndrome coronavirus IgG antibody testing by immunofluorescence antibody assay and confirmation by neutralization test. Parents of all of the subjects who joined the study were contacted by telephone, and a standardized questionnaire was administered by a research nurse to collect information including sociodemographic data, history of severe acute respiratory syndrome coronavirus infection in the subjects and members of the household, history of contact with known cases of severe acute respiratory syndrome, presence of severe acute respiratory syndrome-like symptoms since onset of the severe acute respiratory syndrome epidemic, travel history of the child and his/her relatives within the 15 days before any such symptom onset, use of health service as a result of such symptoms, and whether there were deaths of relatives as a result of severe acute respiratory syndrome. RESULTS Two (0.57%) of 353 asymptomatic children from the high-risk area were tested positive for severe acute respiratory syndrome coronavirus antibody compared with 0 of 361 in the low-risk region. None of the 14 children who lived in the high-risk area and had known contacts with severe acute respiratory syndrome patients were seropositive. CONCLUSIONS As in adults, subclinical severe acute respiratory syndrome coronavirus infection was rare in children in the 2003 epidemic. The very low seroprevalence implies little or no population herd immunity to protect against future resurgence of severe acute respiratory syndrome.


Recombination in RNA viruses and in virus-resistant transgenic plants.


title: Serositis: comparative analysis of histological findings and pathogenetic mechanisms in nonbacterial serosal inflammation.

text: Peritonitis is the established term for infective inflammation of the peritoneum, while serositis generally refers to nonorganismal inflammation in any serous cavity, including the peritoneum. In continuous ambulatory peritoneal dialysis (CAPD) literature, however, culture-negative peritoneal inflammation is referred to as "sterile" or "chemical" peritonitis. These terms not only imply unwarranted etiologic assumptions, but may also deflect attention from the existence of medical conditions to which the peritoneum is subject. This is evident in CAPD literature where there is little recognition that the peritoneum, as a member of the serosa and a secretor of lamellar bodies, is prey to a wide range of disorders. Thus before, during, and after CAPD, the membrane is liable to fall victim to disease states unconnected with the process of dialysis. Significant peritoneal pathology occurs as part of a pan-serositis, which may be metabolic (uremia, cholesterolosis), autoimmune (systemic lupus erythematosus, rheumatoid disease, acute rheumatism, endocrinopathies), genetic (recurrent hereditary polyserositis), allergic (eosinophilic serositis), and granulomatous in nature. This paper presents a comparative analysis of histopathological presentation and pathogenetic mechanisms involved in all forms of peritoneal serositis. It incorporates recent advances in molecular biology of the membrane into a holistic reappraisal of peritoneal pathology, revealing hitherto unrecognized homologies in peritoneal reaction to diverse disorders.


title: Next revolution in the molecular theranostics of infectious diseases: microfabricated systems for personalized medicine.

text: The molecular diagnosis of infectious diseases is currently going through a revolution sustained by the regulatory approval of amplification tests that have been shown to be equivalent or superior to existing gold standard methods. The recent approval of a microarray system for the pharmacogenomic profiling of cytochrome P450-mediated drug metabolism is paving the way to novel, rapid, sensitive, robust and economical microfabricated systems for point-of-care diagnostics, which are utilized closer and closer to the patient's bedside. These systems will enable the multiparametric genetic evaluation of several medical conditions, including infectious diseases. This forecoming revolution will position molecular theranostics in a broader integrated view of personalized medicine, which exploits genetic information from microbes and human hosts to optimize patient management and disease treatment.


title: [Cloning, expression and purification of SARS coronavirus PUMC2 strain nucleocapsid protein].

text: OBJECTIVE To clone, express and purify nucleocapsid protein from SARS coronavirus PUMC2 strain. METHODS According to the published SARS coronavirus genome sequences, the full length cDNA of N protein from SARS coronavirus PUMC2 strain was cloned by RT-PCR and the cDNA was cloned into the pET32a expression vector. The recombinant N protein was expressed in E. coli BL21 (DE3), and purified by Ni(2+)-NTA. RESULTS Prokaryoticly expressed and purified N protein of SARS coronavirus PUMC2 strain was obtained. CONCLUSIONS The SARS coronavirus recombinant N protein obtained by genetic engineering methods can be used for further functional study of SARS coronavirus N protein.


Avian infectious bronchitis.


title: Complete port-accessed lobectomy by the muscle-sparing method.

text: We report the case of a 53-year-old woman who underwent complete port-accessed middle lobectomy by a new technique that preserves all muscles, including the extracostal and intercostal muscles. The operation was performed by using only thoracovideoscopy, and the resected lobe was withdrawn in a pouch through a subxiphoid incision through the substernal route. This complete port-accessed lobectomy is a new technique and is thought to be less invasive than video-assisted lobectomy with minithoracotomy.


title: Remodeling the endoplasmic reticulum by poliovirus infection and by individual viral proteins: an autophagy-like origin for virus-induced vesicles.

text: All positive-strand RNA viruses of eukaryotes studied assemble RNA replication complexes on the surfaces of cytoplasmic membranes. Infection of mammalian cells with poliovirus and other picornaviruses results in the accumulation of dramatically rearranged and vesiculated membranes. Poliovirus-induced membranes did not cofractionate with endoplasmic reticulum (ER), lysosomes, mitochondria, or the majority of Golgi-derived or endosomal membranes in buoyant density gradients, although changes in ionic strength affected ER and virus-induced vesicles, but not other cellular organelles, similarly. When expressed in isolation, two viral proteins of the poliovirus RNA replication complex, 3A and 2C, cofractionated with ER membranes. However, in cells that expressed 2BC, a proteolytic precursor of the 2B and 2C proteins, membranes identical in buoyant density to those observed during poliovirus infection were formed. When coexpressed with 2BC, viral protein 3A was quantitatively incorporated into these fractions, and the membranes formed were ultrastructurally similar to those in poliovirus-infected cells. These data argue that poliovirus-induced vesicles derive from the ER by the action of viral proteins 2BC and 3A by a mechanism that excludes resident host proteins. The double-membraned morphology, cytosolic content, and apparent ER origin of poliovirus-induced membranes are all consistent with an autophagic origin for these membranes.


title: Outbreak of pandemic influenza A (H1N1-2009) in Singapore, May to September 2009.

text: INTRODUCTION The fi rst case of pandemic influenza A(H1N1) was detected in Singapore on 26 May 2009, 1 month after the fi rst cases of novel influenza A(H1N1) was reported in California and Texas in the United States. The World Health Organization declared the fi rst influenza pandemic of the 21st century on 11 June 2009. MATERIALS AND METHODS Confirmed cases notified to the Ministry of Health between 27 May and 9 July 2009 were analysed. Various indicators of influenza activity were monitored throughout the study period. Estimates of the number of cases of H1N1-2009 were made using the number of polyclinic attendances for acute respiratory infection and influenza-like illness and the weekly prevalence of H1N1-2009. RESULTS Cases in Singapore affected mainly young adults, youths and children. By the end of September 2009, it was estimated that at least 270,000 persons had been infected by pandemic influenza A (H1N1) in Singapore. The peak number of cases occurred during E-week 30 (26 July-1 August) when an estimated 45,000 cases were seen in polyclinics and GP clinics. The hospitalisation, severe illness and mortality rates were estimated at 6 per 1000 cases, 0.3 per 1000 cases and 6.7 per 100,000 cases, respectively. The most common risk factors among hospitalised adult cases were asthma and diabetes. For hospitalised children, the most common risk factors were being under 5 years of age and asthma. The most common risk factors among persons with severe illness were diabetes in adults and epilepsy and being under 5 years of age in children. About half of cases with severe illness required mechanical ventilation. In addition, one-fifth of cases with severe illness had acute respiratory distress syndrome. CONCLUSIONS The fi rst wave of the influenza pandemic lasted about 10 weeks. Morbidity and mortality resulting from pandemic influenza were low.


title: Spread of a neurotropic coronavirus to spinal cord white matter via neurons and astrocytes.

text: Mouse hepatitis virus strain JHM (MHV-JHM) causes a chronic encephalomyelitis in susceptible mice, with histological evidence of demyelination in the spinal cord. After intranasal inoculation, virus spreads retrogradely to several brain structures along neuroanatomic projections to the main olfactory bulb. In the absence of experimental intervention, mice become moribund before the spinal cord is infected. In this study, infusions of anti-MHV neutralizing monoclonal antibodies were administered to protect mice from the MHV-JHM-induced acute encephalitis and to allow survival until virus spread to the spinal cord. Under these conditions, virus was observed to enter specific layers (primarily laminae V to VII) in the gray matter of the upper spinal cord, consistent with transneuronal spread. While the brain structures which are the sources for virus spread to the spinal cord cannot be determined with certainty, the ventral reticular nucleus is likely to be important since it is consistently and extensively labeled in all mice and receives projections from subsequently infected areas of the spinal cord. After initial entry into the gray matter, virus rapidly spread to the white matter of the spinal cord. During the early stages of this process, extensive infection of astrocytes was noted, suggesting that cell-to-cell spread via these glial cells is an important part of this process. Reports from other laboratories using cultured cells strongly suggested that astrocytes serve as important regulators of oligodendrocyte function and, by extrapolation, have a major role in vivo in the processes of both demyelination and remyelination. Thus, our results not only outline the probable pathway used by MHV-JHM to infect the white matter of the spinal cord but also, with the assumption that infection of astrocytes leads to subsequent dysfunction, raise the possibility that infection of these cells contributes to the demyelinating process.


title: Disassociation between the in vitro and in vivo effects of nitric oxide on a neurotropic murine coronavirus.

text: Intranasal inoculation of the neuroattenuated OBLV60 strain of mouse hepatitis virus results in infection of mitral neurons in the olfactory bulb, followed by spread along olfactory and limbic pathways to the brain. Immunocompetent BALB/c mice were able to clear virus by 11 days postinfection (p.i.). Gamma interferon (IFN-gamma) may play a role in clearance of OBLV60 from infected immunocompetent BALB/c mice through a nonlytic mechanism. Among the variety of immunomodulatory activities of IFN-gamma is the induction of expression of inducible nitric oxide synthase (iNOS), an enzyme responsible for the production of nitric oxide (NO). Studies were undertaken to investigate the role of IFN-gamma and NO in host defense and clearance of OBLV60 from the central nervous system (CNS). Exposure of OBLV60-infected OBL21a cells, a mouse neuronal cell line, to the NO-generating compound S-nitroso-L-acetyl penicillamine resulted in a significant decrease in viral replication, indicating that NO interfered with viral replication. Furthermore, infection of IFN-gamma knockout (GKO) mice and athymic nude mice with OBLV60 resulted in low-level expression of iNOS mRNA and protein in the brains compared to that of OBLV60-infected BALB/c mice. Nude mice were unable to clear virus and eventually died between days 11 and 14 p.i. (B. D. Pearce, M. V. Hobbs, T. S. McGraw, and M. J. Buchmeier, J. Virol. 68:5483-5495, 1994); however, GKO mice survived infection and cleared virus by day 18 p.i. These data suggest that IFN-gamma production in the olfactory bulb contributed to but may not be essential for clearance of OBLV60 from the brain. In addition, treatment of OBLV60-infected BALB/c mice with aminoguanidine, a selective inhibitor of iNOS activity, did not result in any increase in mortality, and the mice cleared the virus by 11 days p.i. These data suggest that although NO was able to block replication of virus in vitro, expression of iNOS with NO release in vivo did not appear to be the determinant factor in clearance of OBLV60 from CNS neurons.


title: Evaluation of the novel respiratory virus surveillance program: Pediatric Early Warning Sentinel Surveillance (PEWSS).

text: OBJECTIVES Infections caused by respiratory viruses are associated with recurrent epidemics and widespread morbidity and mortality. Routine surveillance of these pathogens is necessary to determine virus activity, monitor for changes in circulating strains, and plan for public health preparedness. The Southern Nevada Health District in Las Vegas, Nevada, recruited five pediatric medical practices to serve as sentinel sites for the Pediatric Early Warning Sentinel Surveillance (PEWSS) program. METHODS Sentinel staff collected specimens throughout the year from ill children who met the influenza-like illness case definition and submitted specimens to the Southern Nevada Public Health Laboratory for molecular testing for influenza and six non-influenza viruses. RESULTS Laboratory results were analyzed and reported to the medical and general communities in weekly bulletins year-round. PEWSS data were also used to establish viral respiratory seasonal baselines and in influenza vaccination campaigns. The surveillance program was evaluated using the Centers for Disease Control and Prevention's (CDC's) Updated Guidelines for Evaluating Public Health Surveillance Systems. PEWSS met three of six program usefulness criteria and seven of nine surveillance system attributes, which exceeded the CDC Guidelines evaluation criteria for a useful and complete public health surveillance program. CONCLUSION We found that PEWSS is a useful and complete public health surveillance system that is simple, flexible, accessible, and stable.


title: Natural West Nile virus infection in a captive juvenile Arctic wolf (Canis lupus).

text: A case of West Nile virus (WNV) infection in a captive 4-month-old Arctic wolf (Canis lupus) is described. The animal had vomiting, anorexia, and ataxia before death. Histopathology revealed multifocal severe renal lymphoplasmacytic vasculitis, mostly affecting small arterioles, with fibrinoid degeneration of some vessel walls. Many small foci of gliosis were detected in the cerebral cortex. West Nile virus was demonstrated in the kidneys and cerebrum by immunohistochemistry and polymerase chain reaction. The described renal changes represent a novel pathological finding of WNV infection.


title: Adult respiratory distress syndrome.

text: ARDS or acute respiratory distress syndrome continues to be a considerable critical care challenge. Mortality has not decreased significantly over the last more than 30 years. This article presents an overview of origin, evaluation, and treatment of ARDS. Recent findings relative to onset and precipitators of ARDS have led to changes in evaluation and treatment plans. Clinical and radiologic descriptors in assessment of the patient with ARDS are discussed. Ventilatory modes and nursing interventions to optimize patient outcomes are identified. The challenges of outcomes issues presented offer opportunities for further study.


Use of humanised mice to study antiviral activity of human γδ-T cells against influenza A viruses.


title: [A cell line particularly susceptible to bovine enteric Coronavirus replication: HRT 18 cells].

text: Bovine enteric Corona virus (BEC) multiplies spontaneously with very high efficiency in HRT 18 cells; this line established from human rectum adenocarcinoma presents some properties of the differentiated brush border cell of the intestinal villi. Titers obtained are very high (5 x 10(7) TCID 50/ml virus strain F15). The virus produced had the characteristic of BEC: shape, spikes, density 1.19, hemadsorption and hemagglutination of rat erythrocytes. Human and dog enteritic corona viruses replicate also in HRT 18 cells.


title: Immunopathogenesis and immunotherapy of multiple sclerosis.

text: Multiple sclerosis (MS) is a chronic disease of the CNS that is characterized by inflammation, demyelination and axonal injury. Although the etiology of MS is still unknown, many findings point toward a central role for the immune system in the pathogenesis of the disease. This hypothesis is strongly supported by the beneficial effects of immunomodulatory and immunosuppressive therapy on disease activity. Over the past few years, substantial progress has been made in deciphering the immune response in MS. Although animal models have advanced our knowledge of basic mechanisms of immune responses in the CNS, recent studies have also highlighted the differences between MS and its animal equivalent, experimental autoimmune encephalomyelitis. New immunotherapeutic agents have been developed and evaluated in clinical trials. Here, we review current knowledge of the immunopathogenesis of MS and corresponding animal models of disease, and discuss new immunointerventional treatment strategies based on changing pathogenetic concepts.


Racial discrimination and mental illness.


title: Molecular characterization and typing of chicken and turkey astroviruses circulating in the United States: implications for diagnostics.

text: Avian astroviruses were detected by reverse transcriptase and polymerase chain reaction in intestinal contents collected from commercial chickens and turkeys from throughout the United States from 2003 through 2005. Astroviruses were detected in birds from both healthy and poorly performing flocks with or without enteric disease. Phylogenetic analysis was performed with sequence data from the polymerase (ORF-1b) genes of 41 turkey-origin astroviruses and 23 chicken-origin astroviruses. All currently available avian astrovirus sequence data and selected mammalian astrovirus sequence data were included in the analysis. Four groups of avian astroviruses were observed by phylogenetic analysis: turkey astrovirus type 1 (TAstV-1)-like viruses, turkey astrovirus type 2 (TAstV-2)-like viruses, both detected in turkeys; avian nephritis virus (ANV)-like viruses, detected in both chickens and turkeys; and a novel group of chicken-origin astroviruses (CAstV). Among these four groups, amino acid identity was between 50.1% and 73.8%, and was a maximum of 49.4% for all avian isolates when compared with the mammalian astroviruses. There were multiple phylogenetic subgroups within the TAstV-2, ANV, and CAstV groups based on 9% nucleotide sequence divergence. Phylogenetic analysis revealed no clear assortment by geographic region or isolation date. Furthermore, no correlation was observed between the detection of a particular astrovirus and the presence of enteric disease or poor performance. Based on these data, a revision of the present taxonomic classification for avian astroviruses within the genus Avastrovirus is warranted.


title: Nucleotide sequence of the gene encoding the spike glycoprotein of human coronavirus HCV 229E.

text: The gene encoding the spike glycoprotein of the human coronavirus HCV 229E has been cloned and sequenced. This analysis predicts an S polypeptide of 1173 amino acids with an Mr of 128,600. The polypeptide has 30 potential N-glycosylation sites. A number of structural features typical of coronavirus S proteins can be recognized, including a signal sequence, a membrane anchor, heptad repeat structures and a carboxy-terminal cysteine cluster. A detailed, computer-aided comparison with the S proteins of infectious bronchitis virus, feline infectious peritonitis virus, transmissible gastroenteritis virus and murine hepatitis virus, strain JHM is presented. We have also done a Northern blot analysis of viral RNAs in HCV 229E-infected cells using synthetic oligonucleotides. On the basis of this analysis, and by analogy to the replication strategy of other coronaviruses, we are able to propose a model for the organization and expression of the HCV 229E genome.


title: [What is the origin of SARS-CoV-2?]

text: Every time a pandemic occurs, dozens of theories emerge to attribute the origin of the event to different facts. The COVID-19 pandemic that has hit virtually all the globe has been no exception. What is known so far about the origin of the virus that causes COVID 19? The first investigations on the origin of this disease have determined that it is a new type of virus, the origin of which is most likely zoonotic.


title: Emergence of a new genotype of avian infectious bronchitis virus in Brazil.

text: Infectious bronchitis virus (IBV) is the agent of a highly contagious disease that affects domestic fowl (Gallus gallus). Recent reports showed a high prevalence of one main IBV genotype (Brazil or BR-I) with low genetic diversity in commercial poultry flocks from Brazil. This research analyzed IBV positive poultry flocks from different rearing regions to verify the S1 gene variability and geographic distribution of variant IBV strains in recent years (2010 and 2011). Samples of IBV-positive flocks were obtained from 60 different farms. Forty-nine partial S1 gene sequences were determined and aligned for phylogenetic and amino acid similarity analyses. Eleven samples (22.4%) were similar to Massachusetts vaccine strains (Mass genotype) and 34 samples (69.4%) to the previously characterized Brazilian BR-I genotype. Interestingly, the remaining four samples (8.2%) clustered into a new IBV variant genotype (Brazil-II or BR-II), divergent from the BR-I. A unique nucleotide sequence insertion coding for five amino acid residues was observed in all the Brazilian variant viruses (BR-I and BR-II genotypes). These results show a higher genetic diversity in Brazilian IBV variants than previously described.


title: Immunotoxicity of monoclonal antibodies.

text: Monoclonal antibodies (mAbs) are large molecules intended to bind to specific targets often expressed on the immune system, and to treat various immunopathological conditions. Therefore, mAbs can be considered to have a high potential for immunotoxicity, which is reflected in the clinical experience accumulated on mAbs-induced adverse effects related to immunosuppression, immunostimulation and hypersensitivity (immunogenicity). So far, non clinical immunotoxicity studies have been inadequate to address all safety issues in relation to the possible immunotoxicity of mAbs, because they are fraught with limitations and pitfalls primarily related to the lack of relevant animal species. In addition, clinical studies rarely include validated end-points dedicated to the prediction of immunotoxicity. With the ongoing development of mAbs as novel therapeutic strategies for a wide variety of diseases, efforts should be paid to improve our understanding of mAbs-induced immunotoxic effects and design dedicated strategies to assess their immunological safety, both non clinically and clinically.


title: The 5'-end sequence of the genome of Aichi virus, a picornavirus, contains an element critical for viral RNA encapsidation.

text: Picornavirus positive-strand RNAs are selectively encapsidated despite the coexistence of viral negative-strand RNAs and cellular RNAs in infected cells. However, the precise mechanism of the RNA encapsidation process in picornaviruses remains unclear. Here we report the first identification of an RNA element critical for encapsidation in picornaviruses. The 5' end of the genome of Aichi virus, a member of the family Picornaviridae, folds into three stem-loop structures (SL-A, SL-B, and SL-C, from the most 5' end). In the previous study, we constructed a mutant, termed mut6, by exchanging the seven-nucleotide stretches of the middle part of the stem in SL-A with each other to maintain the base pairings of the stem. mut6 exhibited efficient RNA replication and translation but formed no plaques. The present study showed that in cells transfected with mut6 RNA, empty capsids were accumulated, but few virions containing RNA were formed. This means that mut6 has a severe defect in RNA encapsidation. Site-directed mutational analysis indicated that as the mutated region was narrowed, the encapsidation was improved. As a result, the mutation of the 7 bp of the middle part of the stem in SL-A was required for abolishing the plaque-forming ability. Thus, the 5'-end sequence of the Aichi virus genome was shown to play an important role in encapsidation.


title: Herpes simplex virus, meningitis and encephalitis in neonates.

text: The consequences of neonatal herpes simplex virus (HSV) infection can be severe. Disease can be localized to skin, eye and mouth (SEM disease), involve the central nervous system (CNS) or manifest as disseminated infection involving multiple organs. Most surviving infants in the latter two categories have neurological sequelae, and the mortality rate in the absence of therapy is very high (80%) for babies in the latter category. The International Herpes Management Forum (IHMF) has produced guidelines on the diagnosis, prevention and effective management of neonatal herpes. Neonatal herpes may occur in the absence of skin lesions, so if the infection is suspected, swabs of the oropharynx, conjunctiva, rectum, skin lesions, mucosal lesions and urine should be promptly taken and submitted for virus culture. Cerebrospinal fluid (CSF) should be submitted for polymerase chain reaction (PCR) detection of HSV DNA. Evidence for disseminated or CNS infection should be sought using liver function tests, complete blood cell count, CSF analysis and chest X-ray, if respiratory abnormalities are present. Neonates with suspected HSV infection should be treated with intravenous aciclovir (20 mg/kg) every 8 h for 21 days. If disease is localized to the SEM, treatment should be limited to 14 days. The neutrophil count for children receiving intravenous aciclovir should be monitored. If the absolute neutrophil count falls below 500/mm3, decreasing the aciclovir dose or administering granulocyte colony stimulating factor (GCSF) should be considered. At the end of therapy in CNS and disseminated disease, PCR assessment of CSF should be used and treatment continued if the child remains PCR positive at this site.


title: Morphologic changes in the nasal cavity associated with sialodacryoadenitis virus infection in the Wistar rat.

text: A sequential study of lesions of the nasal cavity associated with sialodacryoadenitis virus (SDAV) infection was made in the laboratory rat. Wistar rats were intranasally inoculated with approximately 10(3) TCID50 of the coronavirus SDAV. Transverse sections of four regions of the nasal cavity from inoculated and control animals were examined by light microscopy and immunohistochemistry at 2, 4, 6, 8, 10, and 14 days postinoculation (PI). Lesions were observed in the following regions of the upper respiratory tract: respiratory epithelium, transitional epithelium, olfactory epithelium, nasolacrimal duct, vomeronasal organ, and the submucosal glands of the nasal passages. In general, in structures lined by ciliated epithelial cells, there was focal to segmental necrosis with exfoliation of affected cells and polymorphonuclear cell infiltration during the acute stages, progressing to squamous metaplasia during the reparative stages. Repair in these regions was essentially complete by 14 days PI. In the olfactory epithelium and the vomeronasal organ, there was interstitial edema with necrosis and exfoliation of epithelial cells and minimal to moderate inflammatory cell response during the acute stages. Residual reparative lesions were still evident in the olfactory epithelium, the columnar epithelium and neuroepithelium of the vomeronasal organ, and the nasolacrimal duct at 14 days PI. Viral antigen was demonstrated by immunohistochemistry in all regions during the acute stages of the disease, with the exception of the vomeronasal organ. In view of these findings, infections of the respiratory tract with viruses such as SDAV could have significant effects on functions such as olfaction and chemoreception for > or = 2 weeks postexposure in this species.


title: Treatment of pediatric diaphyseal femur fractures.

text: Methods of treating pediatric diaphyseal femur fractures are dictated by patient age, fracture characteristics, and family social situation. The recent trend has been away from nonsurgical treatment and toward surgical stabilization. The clinical practice guideline on pediatric diaphyseal femur fractures was undertaken to determine the best evidence regarding a number of different options for surgical stabilization. The recommendations address treatments that include Pavlik harness, spica casts, flexible intramedullary nailing, rigid trochanteric entry nailing, submuscular plating, and pain management. The guideline authors conclude that controversy and lack of conclusive evidence remain regarding the different treatment options for pediatric femur fractures and that the quality of scientific evidence could be improved for the revised guideline.


title: Australian Hajj pilgrims' knowledge, attitude and perception about Ebola, November 2014 to February 2015.

text: Upon return from Hajj 2014, 150 Australian pilgrims were interviewed about their understanding of the Ebola epidemic. Most (89%, 134/150) knew of the epidemic before travelling and 60% (80/134) of those knew Ebola transmits through body fluids. Pilgrims who received pre-travel health advice were more conscious of Ebola (69% vs 31%, p = 0.01) and adhered better to hand hygiene after touching an ill person (68% vs 31%, p < 0.01). Mass media was the main information source (78%).


title: The threat and prospects for control of an influenza pandemic.

text: Influenza constitutes the most widespread and significant respiratory infectious disease in the world, resulting in increased morbidity, mortality and economic loss each epidemic year. Pandemic influenza is a worldwide epidemic usually caused by a new virus variant to which the majority of the population has no immunity. As demonstrated in the devastating pandemic of 1918 to 1919, a pandemic virus may infect 30 to 50% of the worlds population and kill 1 to 2% of those infected. Pandemic control must be a concerted and co-ordinated world strategy and under the auspices of the World Health Organization, pandemic preparedness plans have been formulated, including: intensified surveillance for more rapid identification of new reassortant viruses with potential human virulence and infectivity, laboratory characterization of the new viruses so that vaccine may be prepared, development of techniques for more rapid vaccine production and the manufacture and stock piling of antiviral drugs. The H5N1 outbreak of virulent chicken influenza in 1997 in Hong Kong which resulted in the deaths of six of 18 infected persons serves as a wake-up call. Should such a virus attain high transmissibility in humans, a pandemic of tragic proportions might ensue. Even though the timing of onset of the next pandemic cannot be precisely predicted, world governments must understand the urgency of the problem and increase funding for influenza pandemic control.


title: [Monitoring of influenza and other respiratory diseases causative agents in children, hospitalized with community-acquired pneumonia in 2012-2013 epidemic season].

text: AIM Study the circulation of respiratory viruses in children with community-acquired pneumonia (CAP) during the period from October 2012 to May 2013. MATERIALS AND METHODS 136 children with CAP aged from 3 months to 16 years with ARI symptoms at the disease debut were studied. RNA/DNA of influenza A, B, parainfluenza (PI); adeno-, rhino-, RS-viruses, corona-, metapneumo- (MPV) and bocaviruses were detected in nasopharynx smears by PCR with hybridization-fluorescent detection in real time. Antibodies against influenza viruses A/H1N1/pdm09 California/07/09, epidemic reference strains of influenza viruses A/H1N1/Brisbane/59/07, A/ H3N2/Victoria/361/201 1, B/Wisconsin/1/10, against PI viruses type 1, 2, 3 were determined in paired sera by HAI. RESULTS In February-March 2013 the number of children protected by antibodies against influenza decreased, and circulation of influenza viruses A/H3N2 and A/H1N1/ pdm09 was detected. Rhinoviruses and PI viruses were determined throughout the epidemic season, bocavirus and adenoviruses--during the autumn-winter period, RS-virus and MPV--during winter-spring. Coronaviruses were not detected. The peak of virus detection was established in February when the threshold of influenza and ARI morbidity was exceeded. The main pathogens of children of the first 3 years of life are rhinoviruses, RS-virus, PI viruses and bocavirus. RS-virus infection at the debut of CAP in children younger than 3 years in 55.5% of cases is associated with the development of broncho-obstructive syndrome. Bocavirus infection in 50% of cases progresses with laryngo-tracheitis and bronchiolitis. CONCLUSION The fraction of viruses in etiologic structure ofARI in children varies depending on immune layer, season and age of children. Etiology of viral infection at the debut of CAP could only be proven using specialized laboratory studies.


title: Assessment of the inhibitory effect of ribavirin on the rainbow trout rhabdovirus VHSV by real-time reverse-transcription PCR.

text: Viral hemorrhagic septicemia virus (VHSV) is one of the most ubiquitous viruses in salmonid aquaculture in Europe. This infectious disease results in significant losses in the farming industry and therefore effective therapeutic agents are needed to control outbreaks caused by this pathogen. Thus, accurate methods to test new antiviral compounds need to be developed. Our goal was to establish a model system for testing novel antivirals with potential applications to aquaculture. In a previous study, a TaqMan real-time RT-PCR assay was designed to detect and quantitate VHSV in rainbow trout tissues [Chico, V., Gomez, N., Estepa, A., Perez, L., 2006. Rapid detection and quantitation of viral hemorrhagic septicemia virus in experimentally challenged rainbow trout by real-time RT-PCR. J. Virol. Methods 132, 154-159]. In this report, we applied the real-time RT-PCR assay to the evaluation of the inhibitory effect of ribavirin, a well-known broad spectrum antiviral drug, in a cell culture system. When added from the beginning of the infection, ribavirin caused a dose-dependent reduction of VHSV RNA accumulation. Real-time RT-PCR measurements showed 99.8% inhibition at 25 microg/ml ribavirin, with an IC50 of 0.43 microg/ml. Ribavirin maintained its inhibitory activity against VHSV when added at 6 h post-infection. Quantitation of N protein messenger RNA and plus-stranded RNA showed a substantial decrease of viral transcription in ribavirin-treated cells. Partial reversion of the effect of ribavirin by addition of GTP was observed, confirming that ribavirin targets the synthesis of guanidine nucleotides in the cells. This is the first report of a real-time PCR-based assay for addressing the efficacy and mechanism of action of an antiviral agent for rainbow trout.


title: [Evaluation of noninvasive positive pressure ventilation in treatment for patients with severe acute respiratory syndrome].

text: OBJECTIVE To analyze the effects of noninvasive positive pressure ventilation (NIPPV) on oxygenation of severe acute respiratory syndrome (SARS) patients, and to discuss the timing point for mechanical ventilation. METHODS Twenty-five SARS patients with respiratory dysfunction treated with NIPPV were studied retrospectively in order to evaluate the influences within 24 hours after initiation of ventilatory support on their physiological indices and oxygenation. Patients with SARS were divided into two groups: survivor group (n=13) and non-survivor group (n=12). We compared the acute physiology and chronic health evaluation (APACHEII) score, respiratory rate (RR),saturation of oxygen (SpO(2)) and modificative respiratory index (MRI) for the survivors and non-survivors before NIPPV and after NIPPV for twenty-four hours, respectively. RESULTS Although NIPPV administered via full-face masks might be an effective treatment for rapidly improving vital signs and gas exchange and sense of dyspnea in both groups during the initial 24 hours of ventilatory support, the patients in non-survivor group had higher APACHEII score, respiratory rates and lower SpO(2), MRI than the patients in survivor group (P<0.05) at the same intervals after initiation of support. CONCLUSION Noninvasive ventilation should be used as a substitutive tool for endotracheal intubation an alternative treatment for acute respiratory failure related to SARS. Therefore, we should make efforts to avoid missing the time point for NIPPV or intubation, and we should not be restricted to the available indications for NIPPV or IPPV.


title: Protection from infection with severe acute respiratory syndrome coronavirus in a Chinese hamster model by equine neutralizing F(ab')2.

text: To warrant potential clinical testing, the equine anti-severe acute respiratory syndrome coronavirus (SARS-CoV) F(ab')(2) requires evaluation in as many animal models as possible. In this study, we established a new animal model, the Chinese hamster, susceptible to SARS-CoV infection. SARS-CoV could propagate effectively and sustain high levels for 1 wk in animal lungs. All animals were protected from SARS-CoV infection in preventive settings. Further, when used therapeutically this antibody led to an approximately 4-log(10) decrease in viral burden in infected animal lungs. The pathological changes in lungs correlated closely with the dose of antibody administered. The excellent preventive and therapeutic roles of equine anti-SARS-CoV F(ab')(2) in several animal models, including the novel Chinese hamster model described in this study, have provided exciting data concerning its potential clinical study.


title: A novel glycoprotein of feline infectious peritonitis coronavirus contains a KDEL-like endoplasmic reticulum retention signal.

text: A new protein of feline infectious peritonitis coronavirus (FIPV) was discovered in lysates of [35S]cysteine-labeled infected cells. Expression of open reading frame (ORF) 6b of FIPV in recombinant vaccinia virus-infected cells was used to identify it as the 6b protein. Further characterization revealed that it is a novel type of viral glycoprotein whose function is not clear. It is a soluble protein contained in microsomes; its slow export from the cell is caused by the presence of an endoplasmic reticulum (ER) retention signal at the C terminus. This amino acid sequence, KTEL, closely resembles the consensus KDEL signal of soluble resident ER proteins. A mutant 6b protein with the C-terminal sequence KTEV became resistant to digestion by endo-beta-N-acetylglucosaminidase H with a half-time that was reduced threefold. In contrast, a mutant with the sequence KDEL was completely retained in the ER. The FIPV 6b protein is the first example of a viral protein with a functional KDEL-like ER retention signal.


title: Interim results: state-specific seasonal influenza vaccination coverage - United States, August 2009-January 2010.

text: The advent of the 2009 influenza A (H1N1) pandemic in April 2009 made the 2009-10 influenza season highly unusual. Public awareness of the potential seriousness of influenza was heightened by media coverage of pandemic-associated hospitalizations and deaths, especially among younger persons. In the fall, the distribution of two separate influenza vaccines began, with distinct, although overlapping, recommendations from the Advisory Committee on Immunization Practices (ACIP). In addition, 2009-10 was the first full season in which ACIP's recommendation to vaccinate all children aged 5--18 years was implemented. To provide preliminary state-specific estimates of seasonal influenza vaccination coverage, CDC analyzed Behavioral Risk Factor Surveillance System (BRFSS) and National 2009 H1N1 Flu Survey (NHFS) data collected during October 2009-February 2010. By January 31, estimated state seasonal influenza vaccination coverage among persons aged >or=6 months ranged from 30.3% to 54.5% (median: 40.6%). Median coverage was 41.2% for children aged 6 months-17 years, 38.3% for adults aged 18--49 years with high-risk conditions, 28.8% for adults aged 18-49 years without high-risk conditions, 45.5% for adults aged 50-64 years, and 69.3% for adults aged >or=65 years. These results, compared with the previous season, suggest large increases in coverage for children and a moderate increase for adults aged 18-49 years without high-risk conditions. Health departments should identify best practices that lead to higher vaccination coverage and should support effective vaccination services (e.g., school-located vaccination programs and office-based protocols, such as reminder/recall and standing orders).


title: Serological studies in Austrian fattening pigs with respiratory disorders.

text: Serum samples of 253 fattening pigs out of 44 different herds with respiratory signs were examined for antibodies to Aujeszky's disease virus by ELISA, swine influenza virus (HI test), porcine respiratory coronavirus (ELISA) and porcine reproductive and respiratory syndrome virus (IPMA). One single case of Aujeszky's disease was detected at slaughter. On the other hand 24.5% of the animals proved to be positive for swine influenza, although no vaccine is licensed in Austria, and 63.6% reacted positively to porcine respiratory coronavirus. None of the sera showed antibodies to porcine reproductive and respiratory syndrome virus, which has not as yet been identified in the Austrian swine population.


docid: eezmtpks

title: Serologic survey of canine coronavirus in wild coyotes in the western United States, 1972-1982.


title: [Clinical application of radionuclide bone scintigraphy in diagnosis of avascular osteonecrosis in patients with severe acute respiratory syndrome in convalescence].

text: OBJECTIVE To evaluate the clinical value of radionuclide bone scintigraphy in diagnosis of avascular osteonecrosis in patients with severe acute respiratory syndrome (SARS) in convalescence. METHODS We performed three-phase bone scintigraphy of femoral head regions and whole-body bone scan in SARS patients 4-6 months after they recovered from the syndrome, and then compared the results with simultaneous MRI. RESULTS Typical avascular necrosis at different stages and severities was found on bone scintigraphy at 31 femoral heads of 16 SARS patients, 97% of which were MRI positive. Suspicious necrosis was found at 42 femoral heads of 23 patients, 67% of which were MRI negative. Among 30 patients with normal three-phase scintigraphic results, 10% of whom were suspicious on MRI. In addition, abnormal distributions of radioactivity were observed in other bones on the whole-body bone scans of 29 patients, including osteonecrosis of knees in 15 patients. CONCLUSIONS Radionuclide bone scintigraphy is valuable in early diagnosis of osteonecrosis in SARS patients in convalescence. It provides a mutually supplementary tool for MRI.


title: Single incision laparoscopic cholecystectomy--a new advantage of gallbladder surgery.

text: In this study is demonstrated our experience in single incision laparoscopic cholecystectomy (SILS), compared to standard laparoscopic cholecystectomy. There were 48 single incision laparoscopic cholecystectomies (SILS) performed during one-year period (A group) and results have been compared with a group of 50 patients who underwent standard laparoscopic cholecystectomy (B group). Outcome measures included operative time, need for conversion, complications, additional analgesia for pain control after procedure, hospital stay and cosmetic outcome. The mean operative time was 46 +/- 3.5 min in A group, and 43 +/- 4 min in B patients group. Early postoperative complications were not detected. The mean hospitalization period was 2 days in both groups. Our experience suggests that SILS cholecystectomy can be performed with outcome similar to standard laparoscopic surgery while affording better cosmesis.


SARS could still affect the United Kingdom, health secretary warns.


title: Identification and characterization of fully human anti-CD22 monoclonal antibodies.

text: CD22 is a member of the B cell receptor family and is implicated in B cell function and development. It is expressed on multiple forms of B cell lymphoma and is an attractive cancer therapeutic target. We report here the identification of two fully human anti-CD22 antibodies using phage display methodology. Both antibodies exhibit specific binding to cell surface-associated CD22 in multiple B cell lines. Through ELISA using mammalian cell-expressed sub-domains of CD22 as binding antigen, we mapped the binding epitopes of the newly identified CD22 antibodies to be within the Ig-like domains 5 to 7 of CD22. Their epitopes do not overlap with those of several therapeutic antibodies currently in preclinical or clinical development. These antibodies have potential as cancer therapeutic candidates and research reagents.


title: Human infection by avian influenza A H5N1.

text: The Southeast Asian outbreak of the highly lethal avian influenza A H5N1 infection in humans is unlikely to abate because of the enormous number of backyard farms providing poultry as the main source of food protein in developing countries. This increases the risk of the emergence of a reassortant pandemic influenza virus with improved human-to-human transmissibility. Currently triage of suspected cases by epidemiological risk factors remains the only practical way of case identification for laboratory investigation and infection control. The clinical usefulness of rapid diagnostic laboratory tests requires more vigorous evaluation. The lethality of this disease may reflect systemic viral dissemination, cytokine storm, or alveolar flooding due to inhibition of cellular sodium channels. The present circulating genotype Z is intrinsically resistant to amantadine and rimantadine. Prognosis may be improved by early treatment with a neuraminidase inhibitor with good systemic drug levels, and post-exposure prophylaxis for health care workers is recommended. The role of immunomodulators and other modalities of therapy requires evaluation in randomised controlled trials, with prospective monitoring of the viral load and cytokine profiles in various clinical specimens. In view of the high fatality of the disease, a combination of contact, droplet, and airborne precautions are recommended as long as resources allow despite the fact that the relative importance of these three modes in nosocomial transmission of avian influenza is still unknown.


title: Mutagenic analysis of the coronavirus intergenic consensus sequence.

text: Previously, a system in which an intergenic region from mouse hepatitis virus (MHV) inserted into an MHV defective interfering (DI) RNA led to transcription of a subgenomic DI RNA in helper virus-infected cells was established. In the present study, a DI cDNA containing one UCUAAAC consensus sequence in the middle of the 0.3-kb-long intergenic region located between genes 6 and 7 was constructed. From this DI cDNA clone, 21 mutant DI RNAs were constructed so that each of the seven consensus sequence nucleotides was changed individually to the three alternative bases. These mutants were used to define how changes in the integrity of MHV transcription consensus sequence UCUAAAC affected mRNA transcription. Except for two mutants with the sequences UGUAAAC and UCGAAAC, all of the mutants supported efficient subgenomic DI RNA transcription. This indicated that MHV transcription regulation was sufficiently flexible to recognize altered consensus sequences. Next, these and other mutants were used to examine the leader-body fusion site on the subgenomic DI RNAs. Sequence analysis demonstrated that all subgenomic DI RNAs analyzed contained two pentanucleotide sequences; the first sequence seemed to be contributed by the leader, and the leader-body fusion most likely took place at either the first or the second nucleotide of the second sequence. This observation was not consistent with the proposed coronavirus transcription model (S. C. Baker and M. M. C. Lai, EMBO J. 9:4173-4179, 1990) which states that nucleotide mismatch can be corrected by RNA polymerase proofreading activity.


Continuous transcutaneous monitoring.


title: Drug targets for rational design against emerging coronaviruses.

text: The recent, fatal outbreak of the novel coronavirus strain in the Middle East highlights the real threat posed by this unique virus family. Neither pharmaceutical cures nor preventive vaccines are clinically available to fight against coronavirus associated syndromes, not to mention a lack of symptom soothing drugs. Development of treatment options is complicated by the unpredictable, recurring instances of cross-species viral transmission. The vastly distributing virus reservoir and the rapid rate of host-species exchange of coronavirus demands wide spectrum potency in an ideal therapeutic. Through summarizing the available information and progress in coronavirus research, this review provides a systematic assessment of the potential wide-spectrum features on the most popular drug targets including viral proteases, spike protein, RNA polymerases and editing enzymes as well as host-virus interaction pathways associated with coronaviruses.


title: [Detection and sequence analysis of an imperfect stem-loop structure in cis activating gene element from SV40PolyA].

text: Our previous studies showed that tandem Alu repeats inhibited GFP gene expression when they were inserted into the downstream of GFP gene in pEGFP-C1 vector and HeLa cells were then transfected transiently. The sequence named 2F2R (second 60 bp from the 5' end of SV40PolyA antisense strand) eliminated the repression of GFP gene expression induced by Alu repeats when 2F2R was inserted between GFP and Alu repeats. In this study the deletion of 2F2R DNA showed that 45R (45 bp in 2F2R 5'end), 30R (30 bp in 2F2R 5' end) and 22R (22 bp in 2F2R 5' end) activated GFP gene expression, and the activating actions of the double tandem sequences were stronger than those of their corresponding single sequences. Secloop (22 bp near the center in 2F2R) and Poly4 (30 bp in 2F2R 3' end) sequences did not activate GFP gene expression. The activating action of 30R-Poly4 sequence formed by ligating 30R with Poly4 by 9 bp was lower than that of 2F2R. The linking base number between two 22R sequences did not influence the GFP gene expression obviously. Sequence 22R (5'-GTGAAAAAAATGCTTTATTTGT-3') contains an imperfect palindrome sequence and may form an imperfect stem-loop structure including a 3nt loop, 3 bp first stem, 2nt bulge, and 3bp second stem. The mutations changing stem-loop structure of 22R influenced the GFP gene activation significantly and neither the excessively stable nor excessively unstable stem-loop structures were in favour of GFP gene activation, which suggested that the suitably imperfect stem-loop structures had something with gene activation.


An iatrogenic pandemic of panic.


title: [COVID-19: patient zero in the Netherlands].

text: BACKGROUND Since December 2019, the world is captivated by SARS-CoV-2, a new coronavirus that shows a lot of similaritieswith previous coronaviruses such as SARS and MERS. Although it was initially seen mainly in China and the surrounding countries, now it also reached Europe, where a large region in northern Italy, in particular, encountered many infections. CASE DESCRIPTION Here we describe the first Dutch patient with COVID-19, a 56-year-old man whose infection appeared to be related to a trip to Northern Italy one week before presentation. In the days that followed, the brother of the patient with whom he had traveled, his wife and daughter also tested positive. CONCLUSION At the moment much is still unclear and it is particularly important to quickly identify patients with an increased risk of complications and to prevent unrestrained spread in the Netherlands.


Heroes and heroines of the war on SARS.


title: [Syndromic surveillance: a finger on the pulse of public health].

text: Syndromic surveillance is a monitoring methodology in which data from disease indicators are collected and interpreted as quickly as possible (in real time) to get an impression of the current morbidity in the population. In this case 'syndrome' means: behavior, signs and symptoms that may be indicative of an infection, without a proven pathogen, or incidents such as an environmental disaster. Syndromic surveillance has gained momentum in response to various international infectious disease threats, including the outbreak of severe acute respiratory syndrome (SARS) in 2003. In several European countries, syndromic surveillance has been used to monitor the impact of outbreaks or natural disasters and the health of visitors of mass events. Following the outbreak of an infectious disease, syndromic surveillance can be reassuring in the absence of an epidemic. Insight into burden of disease can provide guidance in adjusting care capacity to need. Several syndromic surveillance systems and pilot studies have been set up recently in the Netherlands. The costs, benefits and their best application should be evaluated in the near future.


title: Proportion of workers who were work-injured and payment by workers' compensation systems - 10 states, 2007.

text: Work-related injuries are a major cause of morbidity in the United States, with approximately 4 million employer-reported nonfatal injuries and illnesses in 2007. Currently, few population-based state-level estimates of nonfatal occupational injury rates exist. In the few extant studies, self-reported, nonfatal occupational injury rates exceed estimates based on employer reports or state workers' compensation systems. To estimate the proportion of workers who were work-injured during the preceding 12 months and the proportion of those injured for whom workers' compensation insurance programs paid for medical care, 10 states added a module to their 2007 Behavioral Risk Factor Surveillance System (BRFSS) survey. This report summarizes the results of that survey, which found that the proportion of workers who were work-injured during the preceding 12 months ranged from 4.0 to 6.9 work-injured persons per 100 employed persons (Kentucky and New York, respectively). The proportion of self-reported work-injured persons for whom medical treatment was paid by workers' compensation insurance ranged from 47% in Texas to 77% in Kentucky. This study shows the feasibility of complementing existing occupational injury surveillance through the use of population-based surveys. States that wish to enhance existing occupational injury surveillance should consider similar studies. Additional research is needed to understand the reasons for nonpayment of worker-reported occupational injuries by workers' compensation insurance programs.


title: Vaccinia virus leads to ATG12–ATG3 conjugation and deficiency in autophagosome formation.

text: The interactions between viruses and cellular autophagy have been widely reported. On the one hand, autophagy is an important innate immune response against viral infection. On the other hand, some viruses exploit the autophagy pathway for their survival and proliferation in host cells. Vaccinia virus is a member of the family of Poxviridae which includes the smallpox virus. The biogenesis of vaccinia envelopes, including the core envelope of the immature virus (IV), is not fully understood. In this study we investigated the possible interaction between vaccinia virus and the autophagy membrane biogenesis machinery. Massive LC3 lipidation was observed in mouse fibroblast cells upon vaccinia virus infection. Surprisingly, the vaccinia virus induced LC3 lipidation was shown to be independent of ATG5 and ATG7, as the atg5 and atg7 null mouse embryonic fibroblasts (MEFs) exhibited the same high levels of LC3 lipidation as compared with the wild-type MEFs. Mass spectrometry and immunoblotting analyses revealed that the viral infection led to the direct conjugation of ATG3, which is the E2-like enzyme required for LC3-phosphoethanonamine conjugation, to ATG12, which is a component of the E3-like ATG12–ATG5-ATG16 complex for LC3 lipidation. Consistently, ATG3 was shown to be required for the vaccinia virus induced LC3 lipidation. Strikingly, despite the high levels of LC3 lipidation, subsequent electron microscopy showed that vaccinia virus-infected cells were devoid of autophagosomes, either in normal growth medium or upon serum and amino acid deprivation. In addition, no autophagy flux was observed in virus-infected cells. We further demonstrated that neither ATG3 nor LC3 lipidation is crucial for viral membrane biogenesis or viral proliferation and infection. Together, these results indicated that vaccinia virus does not exploit the cellular autophagic membrane biogenesis machinery for their viral membrane production. Moreover, this study demonstrated that vaccinia virus instead actively disrupts the cellular autophagy through a novel molecular mechanism that is associated with aberrant LC3 lipidation and a direct conjugation between ATG12 and ATG3.