Expression, purification and crystallization of the ectodomain of the envelope glycoprotein E2 from Bovine viral diarrhoea virus

Bovine viral diarrhoea virus (BVDV) is an economically important animal pathogen which is closely related to Hepatitis C virus. Of the structural proteins, the envelope glycoprotein E2 of BVDV is the major antigen which induces neutralizing antibodies; thus, BVDV E2 is considered as an ideal target for use in subunit vaccines. Here, the expression, purification of wild-type and mutant forms of the ectodomain of BVDV E2 and subsequent crystallization and data collection of two crystal forms grown at low and neutral pH are reported. Native and multiple-wavelength anomalous dispersion (MAD) data sets have been collected and structure determination is in progress.

Efficacy of Microwaves and Chlorhexidine on the Disinfection of Pacifiers and Toothbrushes: An In Vitro Study

Purpose: The purpose of this study was to evaluate, in vitro, the contamination of toothbrushes and pacifiers by Streptococcus mutans, and the efficacy of microwave and chlorhexidine for their disinfection. Methods: Sixty pacifiers and 60 toothbrushes were contaminated with S mutans and then divided into groups according to the disinfection protocol: Group 1-chlorhexidine solution; Group 2-microwave sterilization; and Group 3-sterile tap water. The devices were evaluated microbiologically as to the formation of S mutans colonies/biofilms and were examined by scanning electron microscopy. The results were submitted for statistical analysis by Friedman`s test at a 5% significance level. Results: The results of both types of evaluation showed a large number of S mutans colonies/biofilms after spraying with sterile tap water, and chlorhexidine spraying and microwaving were effective in eliminate colonies/biofilms. Groups 1 and 2 were statistically similar to each other (P>.05) and differed significantly from Group 3 (P<.05). Conclusions: The 0.12% chlorhexidine solution spray and 7 minutes of microwave irradiation were effective for disinfection of pacifiers and toothbrushes. (Pediatr Dent 2011;33:10-3) Received July 29, 2009 I Last Revision January 26, 2010 I Accepted March 10, 2010

Relation of pretreatment sequence diversity in NS5A region of HCV genotype 1 with immune response between pegylated-INF/ribavirin therapy outcomes

Hepatitis C virus (HCV) infection frequently persists despite substantial virus-specific immune responses and the combination of pegylated interferon (INF)-alpha and ribavirin therapy. Major histocompatibility complex class I restricted CD8+ T cells are responsible for the control of viraemia in HCV infection, and several studies suggest protection against viral infection associated with specific HLAs. The reason for low rates of sustained viral response (SVR) in HCV patients remains unknown. Escape mutations in response to cytotoxic T lymphocyte are widely described; however, its influence in the treatment outcome is ill understood. Here, we investigate the differences in CD8 epitopes frequencies from the Los Alamos database between groups of patients that showed distinct response to pegylated alpha-INF with ribavirin therapy and test evidence of natural selection on the virus in those who failed treatment, using five maximum likelihood evolutionary models from PAML package. The group of sustained virological responders showed three epitopes with frequencies higher than Non-responders group, all had statistical support, and we observed evidence of selection pressure in the last group. No escape mutation was observed. Interestingly, the epitope VLSDFKTWL was 100% conserved in SVR group. These results suggest that the response to treatment can be explained by the increase in immune pressure, induced by interferon therapy, and the presence of those epitopes may represent an important factor in determining the outcome of therapy.

O direito à saúde: a medida do dever do estado de fornecer Medicamentos de dispensação excepcional à sociedade sob a ótica do desenvolvimento

It is a fact that the fundamental rights of citizens are being recognized and guaranteed by the state over time, regardless of the belief that if these rights has always been part of the heritage of subjective individuals, or whether they will be aggregated during the course of human history. In that, emerged the rights of freedom of men and, subsequently, the rights to create a situation of equality between the humans, the so-called social rights. In turn, as these rights known as social, to be implemented, need a positive action by the state, more precisely by the state power whose function is to manage public money and create policies for implementation of fundamental rights. Given this, pay attention to the right to health, was created the Programa de Medicamentos de Dispensação Excepcional, which aims to provide high-cost medicines to citizens Brazilian carriers of serious diseases, such as Alzheimer's and Mal Hepatitis C. Also on the program, it provides a way which will be mandatory that the drugs will be offered in such situations, and does not include a means of updating the list predicted able to monitor the progress of medicine that have been in the interest of the program. Given that, at present it is necessary to mention the recognition of another fundamental right: the right to development, which is the right of access to positive actions being implemented by the State, which are nothing more than public policy, gender which the Programa de Medicamentos de Dispensação Excepcional is kind. Thus, through the search in legislation and doctrine in relation to the theme, this work has the aim to examine the extent of the state to provide exceptional dispensing of medicines. Specifically, if the State in attention to the right to development and the implementation of the right to health, can really list exhaustively the drugs to be provided by the State, and what are the elements guiding this choice and how to control the same

Estudo clínico e níveis de ansiedade em uma série de casos de líquen plano oral

Oral Lichen Planus (OLP) is a relatively common, cronic inflammatory disease. Its etiopathogenesis is no completely understood and several factors have proposed in attempt to explain the appearance, variety of clinical manifestations and periods of exacerbation and remission of the symptons of the lesions. The objective of the present study was to assess the presence of local factors, systemic diseases and levels of anxiety in patients with OLP, investigating their association with the clinical characteristics of the lesions. The sample consisted of 37 patients with histopathologically confirmed OLP that were submitted to a clinical exam to evaluate the presence of smoking habits, consumption of alcoholic beverages, diabetes mellitus, hypertension and hepatitis C virus (HCV) infection. The existence of skin lesions, as well as the time course, clinical form, symptomatology, number and location of the oral lesions were also registered. The trait anxiety was measured by Spielberger s Stai-Trait Anxiety Inventory (STAI), and to associate the variables we used the chi-square or Fisher s exact test. It was observed that females were the most affected (75%) and the mean age of the patients was 53,3 years. Most cases were non-smoker (97,3%) and none was drinker. Diabetes mellitus and hypertension were present in 10,8% e 16,2% of the sample, respectively and only one patient was HCV-seropositive (2,7%). Moderate levels of anxiety were seen in most cases (78,4%) and 21,6% had elevated levels. The oral lesions persisted in 95% of the cases for a period of 6 months to 13 years. The erosive form was the most prevalent (57,1%) and symptons were reported by 45,7% of the patients. Multiple lesions were frequent (60%), affecting mainly the buccal mucosa, followed by gums e tongue. There was no significant association of the presence of diabetes mellitus, hypertension and levels of anxiety with the clinical form or symptomatology of the oral lesions (p>0,005), despite a trend in patients with hypertension to have erosive lesions. It was concluded that, in the sample studied, moderate levels of anxiety were commonly observed, and the HCV infection apparently is not related to the onset of the OLP. In this study, the presence of diabetes mellitus, hypertension and levels of anxiety seem not to be associated with the clinical characteristics of the lesions of OLP

Relation of pretreatment sequence diversity in NS5A region of HCV genotype 1 with immune response between pegylated-INF/ribavirin therapy outcomes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development of a rapid one-step immunochromatographic assay for HCV core antigen detection

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Application of Factorial Design Experiments to the Development of a Disposable Amperometric DNA Biosensor

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

POLYSEROSITIS IN A PATIENT WITH ACUTE PARACOCCIDIOIDOMYCOSIS AND HEPATOSPLENIC SCHISTOSOMIASIS

A severe case of juvenile paracoccidioidomycosis (PCM), manifested as cholestatic jaundice, lymphnode enlargement and an unusual form of polyserositis, associated with portal hypertension secondary to schistosomiasis, as well as bacteremias caused by E. coli and S. aureus and post-transfusional hepatitis C is reported. Temporary unresponsiveness of in vivo and in vitro cellular immune responses to P. brasiliensis were registered. The authors discuss the possible interference of either agent in the host immune response, thus explaining the severity of PCM in the present case.

Polysiloxane-poly(propylene oxide) hybrid discs as solid phase in anti-HCV detection using a recombinant core protein

In this work, siloxane-poly(propylene oxide) discs (PPO disc) prepared using the sol-gel process were used as solid phase in enzyme-linked immunosorbent assays (ELISA) for the detection of anti-hepatitis C virus (HCV) antibodies. The HCV RNA from serum (genotype 1b) was submitted to the RT-PCR technique and subsequent amplification of the HCV core 408 pb. This fragment was cloned into expression vector pET42a and expressed in Escherichia coli as recombinant protein with glutathione S-transferase (GST). Cell cultures were grown and induced having a final concentration of 0.4 x 10(-3) mol L-1 of IPTG. After induction, the cells were harvested and the soluble fraction was analyzed using polyacrilamide gel 15% showing a band with an approximate molecular weight of 44 kDa, the expected size for this GST-fused recombinant protein. The recombinant protein was purified and continued by immunological detection using HCV-positive serum and showed no cross-reactivity with positive samples for other infectious diseases. An ELISA was established using 1.25 ng of recombinant protein per PPO disc, a dilution of 1: 10,000 and 1:40 for a peroxidase conjugate and serum, respectively, and solutions of hydrogen peroxide and 3,3',5,5'-tetra-methylbenzidine in a ratio of 1: 1. The proposed methodology was compared with the ELISA conventional polystyrene-plate procedure and the performance of the PPO discs as a matrix for immunodetection gave an easy synthesis, good performance and reproducibility for commercial application. (c) 2007 Elsevier B.V. All rights reserved.

Viral infections in workers in hospital and research laboratory settings: a comparative review of infection modes and respective biosafety aspects

Objectives: To compare modes and sources of infection and clinical and biosafety aspects of accidental viral infections in hospital workers and research laboratory staff reported in scientific articles. Methods: PubMed, Google Scholar, ISI Web of Knowledge, Scirus, and Scielo were searched (to December 2008) for reports of accidental viral infections, written in English, Portuguese, Spanish, or German; the authors' personal file of scientific articles and references from the articles retrieved in the initial search were also used. Systematic review was carried out with inclusion criteria of presence of accidental viral infection's cases information, and exclusion criteria of absence of information about the viral etiology, and at least probable mode of infection.Results: One hundred and forty-one scientific articles were obtained, 66 of which were included in the analysis. For arboviruses, 84% of the laboratory infections had aerosol as the source; for alphaviruses alone, aerosol exposure accounted for 94% of accidental infections. of laboratory arboviral infections, 15.7% were acquired percutaneously, whereas 41.6% of hospital infections were percutaneous. For airborne viruses, 81% of the infections occurred in laboratories, with hantavirus the leading causative agent. Aerosol inhalation was implicated in 96% of lymphocytic choriomeningitis virus infections, 99% of hantavirus infections, and 50% of coxsackievirus infections, but infective droplet inhalation was the leading mode of infection for severe acute respiratory syndrome coronavirus and the mucocutaneous mode of infection was involved in the case of infection with influenza B. For blood-borne viruses, 92% of infections occurred in hospitals and 93% of these had percutaneous mode of infection, while among laboratory infections 77% were due to infective aerosol inhalation. Among blood-borne virus infections there were six cases of particular note: three cases of acute hepatitis following hepatitis C virus infection with a short period of incubation, one laboratory case of human immunodeficiency virus infection through aerosol inhalation, one case of hepatitis following hepatitis G virus infection, and one case of fulminant hepatitis with hepatitis B virus infection following exposure of the worker's conjunctiva to hepatitis B virus e antigen-negative patient saliva. of the 12 infections with viruses with preferential mucocutaneous transmission, seven occurred percutaneously, aerosol was implicated as a possible source of infection in two cases, and one atypical infection with Macacine herpesvirus 1 with fatal encephalitis as the outcome occurred through a louse bite. One outbreak of norovirus infection among hospital staff had as its probable mode of infection the ingestion of inocula spread in the environment by fomites.Conclusions: The currently accepted and practiced risk analysis of accidental viral infections based on the conventional dynamics of infection of the etiological agents is insufficient to cope with accidental viral infections in laboratories and to a lesser extent in hospitals, where unconventional modes of infection are less frequently present but still have relevant clinical and potential epidemiological consequences. Unconventional modes of infection, atypical clinical development, or extremely severe cases are frequently present together with high viral loads and high virulence of the agents manipulated in laboratories. In hospitals by contrast, the only possible association of atypical cases is with the individual resistance of the worker. Current standard precaution practices are insufficient to prevent most of the unconventional infections in hospitals analyzed in this study; it is recommended that special attention be given to flaviviruses in these settings. (C) 2011 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Immunosensor based on immobilization of antigenic peptide NS5A-1 from HCV and silk fibroin in nanostructured films

The peptide NS5A-1 (PPLLESWKDPDYVPPWHG), derived from hepatitis C virus (HCV) NS5A protein, was immobilized into layer-by-layer (LbL) silk fibroin (SF) films. Deposition was monitored by UV-vis absorption measurements at each bilayer deposited. The interaction SF/peptide film induced secondary structure in NS5A-1 as indicated by fluorescence and circular dichroism (CD) measurements. Voltammetric sensor (SF/NS5A-1) properties were observed when the composite film was tested in the presence of anti-HCV. The peptide-silk fibroin interaction studied here showed new architectures for immunosensors based on antigenic peptides and SF as a suitable immobilization matrix. © 2013 American Chemical Society.

Acidentes de trabalho com material biológico e o seu significado para os profissionais envolvidos

Pós-graduação em Enfermagem (mestrado profissional) - FMB

Soroepidemiologia do HCV em doadores de sangue na cidade de Imperatriz – MA

Devido à alta mortalidade e, principalmente, a morbidade, as hepatites são um dos mais graves problemas de Saúde Pública, no País e no mundo. Entre elas destacamos a infecção da hepatite C. O número de pessoas que desconhecem que são portadoras do vírus HCV é relevante. Como atualmente a transmissão do HCV por transfusão sanguínea e hemoderivados é rara entre os doadores de sangue, depois da introdução do método de triagem nos centros hemoterápicos, fundamental para detectar a existência de uma possível infecção neste doador. A realidade epidemiológica da hepatite C em Imperatriz necessita de maior conhecimento e planejamento das estratégias de prevenção e assistência aos portadores de HCV, uma vez que não existe uma rede de serviço consolidada para o tratamento, a burocracia é grande para se chegar ao diagnóstico da doença e a sub-notificação dos casos é elevada. Tem como objetivo avaliar a soroprevalência do HCV em candidatos à doação de sangue no município de Imperatriz – MA; assim como analisar o perfil dos candidatos considerados inaptos a doação de sangue no HEMOMAR, nesta cidade; determinar a soroprevalência do Vírus da Hepatite C entre os doadores de sangue no período de 2005 a 2010; realizar o levantamento dos dados epidemiológicos, destacando o gênero e a faixa etária de maior prevalência do vírus da hepatite C; comparar os dados epidemiológicos identificando a procedência dos candidatos soropositivos para o vírus da hepatite C. o estudo é de caráter descritivo transversal, envolvendo doadores de sangue do Centro de Hematologia e Hemoterapia do Estado do Maranhão de Imperatriz - MA. Na distribuição dos doadores de sangue do HEMOMAR, regional de Imperatriz -MA quanto ao sexo, constata-se que, nos anos de 2005 à 2011 o fluxo de doadores caracterizou-se por indivíduos de ambos os sexos,com predominância do masculino (75,01%), quando analisamos o perfil dos candidatos a doadores com sorologia positiva para HCV, observamos que estes também eram a maioria. A faixa etária dos doadores de sangue do HEMOMAR predominante era de 18-29. Os candidatos a doação com sorologia positiva para HCV, foram encontrados 79,17% na situação de casados/união estável. Entre os doadores que foram considerados inaptos a doação, 0,21% apresentou sorologia positiva para HCV. A maioria dos candidatos a doadores com soropositividade para HCV pertencia ao município de Imperatriz. Concluiu-se que é importante lembrar que o processo de triagem clínica e laboratorial diminui riscos de contaminação no processo de transfusão sanguínea. A figura do doador de sangue deve ser sempre valorizada e parabenizada por todos.