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.

Influence of HLA alleles in response to treatment with pegylated interferon-alpha and ribavirin in patients with chronic hepatitis C

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Induction pegylated interferon Alfa-2a and high dose ribavirin do not increase SVR in heavy patients with HCV genotype 1 and high viral loads

Background & Aims Patients infected with hepatitis C virus (HCV) genotype 1, body weight <85 kg, and high baseline viral load respond poorly to standard doses of pegylated interferon (peginterferon) and ribavirin. We evaluated intensified therapy with peginterferon alfa-2a plus ribavirin. Methods This double-blind randomized trial included HCV genotype 1-infected outpatients from hepatology clinics with body weight <85 kg and HCV RNA titer <400,000 IU/mL. Patients were randomized to 180 μg/wk peginterferon alfa-2a for 48 weeks plus 1200 mg/day ribavirin (standard of care) (group A, n = 191) or 1400/1600 mg/day ribavirin (group B, n = 189). Additional groups included 360 μg/wk peginterferon alfa-2a for 12 weeks then 180 μg/wk peginterferon alfa-2a for 36 weeks plus 1200 mg/day ribavirin (group C, n = 382) or 1400/1600 mg/day ribavirin (group D, n = 383). Follow-up lasted 24 weeks after treatment. Results Sustained virologic response rates (HCV RNA level <15 IU/mL at end of follow-up) in groups A, B, C, and D were 38%, 43%, 44%, and 41%, respectively. There were no significant differences among the 4 groups or between pooled peginterferon alfa-2a regimens (A + B vs C + D: odds ratio [OR], 1.08; 95% confidence interval [CI], 0.831.39; P = .584) or pooled ribavirin regimens (A + C vs B + D: OR, 1.00; 95% CI, 0.791.28; P = .974). Conclusions In patients infected with HCV genotype 1 who are difficult to treat (high viral load, body weight <85 kg), a 12-week induction regimen of peginterferon alfa-2a and/or higher-dose ribavirin is not more effective than the standard regimen. © 2010 AGA Institute.

Avaliação do perfil lipídico entre indivíduos com hepatite C

Metabolic profiles correlate with hepatitis C virus (HCV) infection and are prognostic for the viral response. However, little is known about the association between lipid profiles and viral load in chronic patients carrying HCV genotypes 1, 2 and 3. The aim of this study was to investigate the influence of the viremia and viral genotype on lipid metabolism by observing the variations in serum lipoprotein and apolipoprotein B, to assess whether HCV predisposes individuals to lipid imbalance and favors the appearance of vascular complications. A sample group of 150 chronic HCV patients with viral genotypes 1, 2 or 3 and a control group of 20 healthy adults (10 men and 10 women), all aged from 20 to 50 years were studied. The serum lipid profile of the chronic patients was analyzed and compared to that of the control group. The high-density lipoprotein (HDL), very low-density lipoprotein (VLDL) and triglyceride levels of the sample group were lower than those of the control group, while the low-density lipoprotein (LDL) and apolipoprotein B levels of the patients were higher. These differences were more significant in patients carrying genotype 3a. There was a positive correlation between the viremia and the changes in apolipoprotein B levels in patients carrying genotype 1b. It was inferred that the risk of developing vascular complications raised in HCV patients. As 90% of LDL protein is composed of apolipoprotein B, the plasmatic concentration of the latter indicates the number of potentially atherogenic particles. Therefore, the lipid profile monitoring may aid in the diagnosis of hepatic infection severity and equally act as a good prognostic marker.

Clonagem e expressão da proteína E2 no vírus da hepatite C Humana: estudo da interação molecular E2-rLDL in vitro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação da prevalência de imunização contra a hepatite B nos profissionais da secretaria de saúde de Maringá

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Estudo da influência dos genótipos 1 e 3 do vírus da hepatite C sobre os indicadores do metabolismo lipídico em hepatopatas crônicos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Imunossensores à base de filmes nanoestruturados de fibroína da seda - peptídeo antigênico NS5A-1-vanadato de ítrio: európio para detecção de Hepatite C

Pós-graduação em Química - IQ

Diagnostic tests for hepatitis C: Recent trends in electrochemical immunosensor and genosensor analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Pesquisa de genomas virais em primatas não humanos do novo mundo por metagenômica

Pós-graduação em Medicina Veterinária - FMVZ

Conhecimento e atitudes da área da saúde frente ao risco de contaminação por hepatites ao fazer as unhas

Pós-graduação em Saúde Coletiva - FMB

Oral syphilis: report of three cases and characterization of the inflammatory cells

Syphilis is a sexually transmitted infectious disease caused by Treponema pallidum. This study reports 3 cases of syphilis and highlights the importance of identifying oral lesions for its final diagnosis. Case 1: a 48-year-old male patient presented with a bleeding ulcer in the lower lip. Overall clinical examination revealed patchy alopecia and skin target lesions. Case 2: a 61-year-old male patient presented with white spots on the lateral tongue and nodules on the dorsum of the tongue. Overall clinical examination showed erythematous target lesions on the abdomen, forearm, palms of the hand, and soles of the feet. Case 3: a 17-year-old male patient presented with an ulcerated lesion on the tongue and lymph node involvement. The following serologic tests were requested: Venereal Disease Research Laboratory, fluorescent treponemal antibody-absorption, anti-HIV-1 and anti-HIV-2, and anti-hepatitis C virus. An incisional biopsy revealed epithelial hyperplasia associated with intense and diffuse mononuclear inflammatory cell infiltration consisting mainly of plasma cells, in a perivascular and perineural distribution. The final diagnosis in the 3 patients was syphilis. Treatment consisted of 1 weekly dose of penicillin (2.4 million units, intramuscular) for 2 or 3 weeks. Immunohistochemical reactions for XIIIa, CD3, CD20, CD68, CD163, S100, CD1a, CD11c, CD83, CD138, and CD208 were performed. Clinicians should be familiarized with oral syphilis lesions in order to be able to diagnose this emerging infectious disease of variable clinical presentation.

Assessment of health-related quality of life and related factors in patients with chronic liver disease

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Re-treatment of previous non-responders and relapsers to interferon plus ribavirin with peginterferon alfa-2a (40KD), ribavirin +/- amantadine in patients with chronic hepatitis C: randomized multicentre clinical trial

Introduction. A large number of patients with chronic hepatitis C have not been cured with interferon-based therapy. Therefore, we evaluated the efficacy of amantadine combined with the standard of care (pegylated interferon plus ribavirin) in patients who had not responded to or had relapsed after 24 weeks of treatment with conventional interferon plus ribavirin. Material and methods. Patients stratified by previous response (i.e., non-response or relapse) were randomized to 48 weeks of open-label treatment with peginterferon alfa-2a (401(D) 180 pg/week plus ribavirin 1,000/1,200 mg/day plus amantadine 200 mg/day (triple therapy), or the standard of care (peginterferon alfa-2a [40KD] plus ribavirin). Results. The primary outcome was sustained virological response (SVR), defined as undetectable hepatitis C virus RNA in serum (< 50 IU/mL) at end of follow-up (week 72). Among patients with a previous non-response, 12/53 (22.6%; 95% confidence interval [CI] 12.3-36.2%) randomized to triple therapy achieved an SVR compared with 16/52 (30.8%; 95% CI 18.7-45.1%) randomized to the standard of care. Among patients with a previous relapse 22/39 (56.4%; 95% CI 39.6-72.2%) randomized to triple therapy achieved an SVR compared with 23/38 (60.5%; 95% CI 43.4-76.0%) randomized to the standard of care. Undetectable HCV RNA (< 50 IU/mL) at week 12 had a high positive predictive value for SVR. A substantial proportion of non-responders and relapsers to conventional interferon plus ribavirin achieve an SVR when re-treated with peginterferon alfa-2a (40KD) plus ribavirin. Conclusion. Amantadine does not enhance SVR rates in previously treated patients with chronic hepatitis C and cannot be recommended in this setting.

Síntese e atividade anti-HCV, anti-candida e antibacteriana de nitrochalconas

Pós-graduação em Química - IBILCE