A simple and cheaper in house varicella zoster virus antibody indirect ELISA

We have developed a cheaper an simple in house indirect ELISA that uses the live attenuated VZV vaccine as a coating antigen. The alternative ELISA had an agreement of 94% when compared with a commercial VZV ELISA kit. Moreover, our ELISA proved to be more reliable than the kit when assessing true negative samples. By adding a standard serum, we were able to produce results in international units per millilitre. Also, the addition of an extra step with 8M urea allowed the assessment of VZV IgG avidity without excessive costs. The cost per sample to test VZV IgG was 2.7 times cheaper with our ELISA, allowing the testing of many samples without the burden of production of VZV antigen in the laboratory.

Serological, epidemiological and molecular aspects of hepatitis C virus infection in a population from Londrina, PR, Brazil, 2001-2002

Serological, epidemiological and molecular aspects of hepatitis C virus (HCV) infection were evaluated in 183 subjects from Londrina, Paraná, Brazil, and adjacent areas. Serum samples which tested anti-HCV positive by microparticle enzyme immunoassay (MEIA) obtained from eight patients with chronic hepatitis C, 48 blood donors, and 127 patients infected with the human immunodeficiency virus (HIV) were submitted to another enzyme immunoassay (ELISA) and to the polymerase chain reaction (PCR). About 78.7% of samples were also reactive by ELISA, with the greater proportion (70.8%) of discordant results verified among blood donors. A similar finding was observed for HCV-RNA detection by PCR, with 111/165 (67.3%) positive samples, with higher rates among HIV-positive subjects and patients with chronic hepatitis than among blood donors. Sixty-one PCR-positive samples were submitted to HCV genotyping, with 77.1, 21.3 and 1.6% of the samples identified as types 1, 3 and 2, respectively. Finally, analysis of some risk factors associated with HCV infection showed that intravenous drug use was the most common risk factor among HIV/HCV co-infected patients, while blood transfusion was the most important risk factor in the group without HIV infection. The present study contributed to the knowledge regarding risk factors associated with HCV infection and the distribution of HCV genotypes in the population evaluated.

Será que a Definição de Síndroma Metabólica pela International Diabetes Federation Melhora a Capacidade Preditora para Doença Coronária e do Espessamento da Íntima-Média Carotídea?

BACKGROUND: Metabolic syndrome (MS) is associated with increased incidence of diabetes and atherosclerotic complications. The new definition of the International Diabetes Federation (IDF) increases the population with this entity, compared to the NCEP ATP III definition. OBJECTIVES: To study the prevalence of coronary artery disease (CAD) and carotid intima-media thickness (IMT) in patients with and without MS, according to the NCEP ATP III and IDF definitions, and the predictive ability of carotid IMT for CAD. METHODS: We studied 270 consecutive patients admitted for elective coronary angiography due to suspicion of CAD. All patients underwent ultrasound study of the carotid arteries to measure IMT (the highest value between the right and left common carotid arteries was used in the analysis). Coronary stenosis of > or =70% (or 50% for the left main coronary artery) was considered significant. RESULTS: By the ATP III definition, 14% of the patients had MS, and these patients had a higher prevalence of CAD (87% vs. 63%, p = 0.004), but no significant difference was found for carotid IMT (1.03 +/- 0.36 mm vs. 0.95 +/- 0.35 mm, p=NS). With the IDF definition, 61% of the patients had MS; this group was slightly older and included more women. There were no differences in terms of CAD (68% vs. 63%) or carotid IMT (0.97 +/- 0.34 vs. 0.96 +/- 0.39 mm). On multivariate analysis, the ATP III definition of MS predicts CAD (OR 4.76, 95% CI 1.71-13.25, p = 0.003), but the IDF definition does not (OR 1.29, 95% CI 0.74-2.27, p = 0.37). On ROC curve analysis, an IMT of > or = 0.95 mm predicts CAD (AUC 0.66, p < 0.001), with a sensitivity of 52% and specificity of 75%. CONCLUSIONS: The new IDF definition increases the population with MS, decreasing the capacity to predict the presence of CAD. In our population, neither the ATP III nor the IDF definition showed differences in terms of carotid IMT. Carotid IMT can predict CAD, but with only modest sensitivity.

Human vaccinia-like virus outbreaks in São Paulo and Goiás States, Brazil: virus detection, isolation and identification

Since October 2001, the Adolfo Lutz Institute has been receiving vesicular fluids and scab specimens of patients from Paraíba Valley region in the São Paulo and Minas Gerais States and from São Patricio Valley, in the Goiás State. Epidemiological data suggested that the outbreaks were caused by Cowpox virus or Vaccinia virus. Most of the patients are dairy milkers that had vesiculo-pustular lesions on the hands, arms, forearms, and some of them, on the face. Virus particles with orthopoxvirus morphology were detected by direct electron microscopy (DEM) in samples of 49 (66.21%) patients of a total of 74 analyzed. Viruses were isolated in Vero cell culture and on chorioallantoic membrane (CAM) of embryonated chicken eggs. Among 21 samples submitted to PCR using primers for hemagglutinin (HA) gene, 19 were positive. Restriction digestion with TaqI resulted in four characteristic Vaccinia virus fragments. HA nucleotide sequences showed 99.9% similarity with Cantagalo virus, described as a strain of Vaccinia virus. The only difference observed was the substitution of one nucleotide in the position 616 leading to change in one amino acid of the protein in the position 206. The phylogenetic analysis showed that the isolates clustered together with Cantagalo virus, other Vaccinia strains and Rabbitpox virus.

Morbilidade Cardiovascular em Crianças com Infecção pelo Vírus da Imunodeficiência Humana

Na infecção pelo vírus da imunodeficiência humana (VIH) estão descritas alterações cardiovasculares, tanto estruturais como funcionais. Com o objectivo de fazer o rastreio das referidas alterações no propósito de uma intervenção terapêutica precoce, foi efectuada uma avaliação clínica e laboratorial prospectiva em 32 crianças com infecção pelo VIH, com idades compreendidas entre três meses e 13 anos (média = 3.11 +/- 3.51 anos). Em 90% dos doentes identificou-se transmissão perinatal. Vinte e duas crianças (69%) estavam sintomáticas, sendo a sintomatologia moderada em nove e grave em oito. Catorze doentes tinham alterações imunológicas e oito delas apresentavam imunosupressão grave. Vinte e oito crianças (88%)tinham infecção pelo VIH 1 e seis tinham infecção recente pelo Vírus de Ebstein-Barr. Dezanove doentes receberam tratamento com zidovudina e 14 com imunoglobulinas por via endovenosa. Foram detectadas 19 alterações cardiovasculares em 15 doentes (47%), nomeadamente: 11 casos de hipertensão pulmonar por critérios ecocardiográficos, (oito delas tinham infiltrados intersticiais na radiografia do tórax) e quatro casos de disfunção ventricular esquerda que, por isso, iniciaram terapêutica anticongestiva. As restantes anomalias estruturais ou funcionais foram: persistência de canal arterial; hipertrofia do septo interventricular; prolapso da válvula mitral e derrame pericárdico cada uma delas em um caso. No electrocardiograma de superfície quatro crianças apresentaram critérios de hipertrofia ventricular direita, uma tinha hipertrofia ventricular esquerda e duas tinham alterações inespecíficas de repolarização ventricular. Em 14 doentes (44%) verificou-se no ECG de 24 horas taquicardia sinusal com uma frequência cardíaca média superior ao percentil 95. As anomalias cardiovasculares foram mais frequentes nas crianças com estadios mais avançados de doença. Conclusões: As anomalias cardiovasculares são frequentes em crianças com infecção pelo VIH em estadios avançados; a hipertensão pulmonar é a anomalia cardiovascular mais frequentemente detectada e está associada a patologia respiratória crónica ou recorrente; o acompanhamento cardiológico está indicado nas crianças com infecção pelo vírus da imunodeficiência humana.

Estudo da Distribuição de Factores de Risco para o Desenvolvimento de Doença Ocular Aguda em Portadores de Lentes de Contacto

Objectivo: Estudar os factores de risco (FR) associados a complicações clínicas agudas (CCA) numa amostra de portadores de lentes de contacto (LC). Doentes e Métodos: Análise de todos os doentes com CCA associadas ao uso de LC, que, no período de um ano, recorreram ao SU do Centro Hospitalar de Lisboa Central. O estudo focou-se na análise do perfil demográfico e de risco destes doentes. Resultados: Dos 103 doentes referenciados pelo SU foram estudados os que compareceram para seguimento no Departamento de Contactologia (84). A maioria era do sexo feminino, caucasiana e de educação diferenciada. A mediana das idades foi de 29 anos. 15% dos doentes cumpria as horas de uso diário recomendadas e 5% cumpria todas as regras de higiene; 26% dos doentes apresentavam FR oftalmológicos e 58% FR sistémicos. O prescritor das LC foi o médico em 36% e 26% mantinha vigilância médica oftalmológica regular. Comentários: A maioria dos doentes não cumpria as regras de segurança e de higiene do uso de LC, sendo o desconhecimento destas regras elevado. O papel muito reduzido dos médicos oftalmologistas na prescrição de LC e na vigilância dos portadores pode estar na origem do desconhecimento e incumprimento das regras de utilização das LC por parte de alguns doentes.

Hepatitis C in patients co-infected with human immunodeficiency virus. A review and experience of a Brazilian ambulatory

Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) share the same transmission mechanisms. The prevalence of HCV in the HIV-infected population varies from region to region, throughout the world, depending on different exposure factors to both viruses. Co-infection with HIV accelerates the progression of the disease caused by HCV, appears to worsen the progression of the HIV infection and increases HCV transmission. Therefore, clinical management and treatment of HCV is a priority in medical facilities that receive HIV-infected patients. Clinical management of these patients involves specific diagnostic procedures and appropriately trained medical staff. The indication of treatment should meet specific clinical and laboratory criteria. There are a number of drugs currently available to treat hepatitis C in co-infected patients.

Avaliação do Conhecimento sobre a sua Doença em Doentes com Glaucoma seguidos na Consulta de Especialidade de um Hospital Central

Objectivo: Avaliar e comparar com a população geral o nível de conhecimento sobre a doença e o seu tratamento em doentes com glaucoma. Doentes e Métodos: Noventa doentes com glaucoma e 90 doentes sem glaucoma foram entrevistados no departamento de glaucoma e de consulta geral do Centro Hospitalar de Lisboa Central. Foi solicitado o preenchimento de um questionário validado sobre a doença e o seu tratamento. Foram registados dados demográficos. Resultados: Dezoito porcento e 51% dos doentes,respectivamente, com glaucoma e controlo, desconheciam a doença. Em 6 das 22 perguntas, mais de 50% dos doentes responderam acertadamente. Em 16 perguntas o número de respostas correctas dos doentes foi inferior a 50%. Os doentes com glaucoma têm um nível de conhecimentos superior (p=7,9x10-6) ao grupo controlo. O nível de conhecimento é maior quanto maior a duração da doença (p=0,03) e o nível de escolaridade (p=0,0065). Comentários: Os doentes com glaucoma têm um nível de conhecimentos sobre a sua doença superior ao grupo controlo, não obstante ambos os grupos terem um conhecimento insuficiente. Devem ser tomadas medidas de prevenção primária e secundária, com recurso a material educacional, com o objectivo de melhorar os conhecimentos dos doentes e consequentemente a compliance.

Análise da Repercussão de uma Sessão Educativa sobre o Glaucoma no Conhecimento da Doença

Introdução: A adesão à terapêutica antiglaucomatosa é fundamental na redução e controlo da pressão intra-ocular. Os doentes com menor nível de conhecimento sobre a sua doença apresentam taxas de não adesão terapêutica superiores. Este estudo tem como objectivo a avaliação da repercussão de uma sessão de esclarecimento sobre o glaucoma e seu tratamento. Material e métodos: Estudo prospectivo que incluiu 24 doentes seguidos em consulta de glaucoma. Procedeu-se ao preenchimento de um questionário validado, sobre a doença e o seu tratamento. De seguida os doentes assistiram a uma sessão de esclarecimento sobre o glaucoma. O preenchimento do questionário foi repetido após a sessão e ao fim de 1 mês. Resultados: A idade média foi 63,7 anos, 45,8% era do sexo feminino. Em 9 das 22 perguntas, mais de 50% dos doentes responderam acertadamente antes da sessão educativa. Após a realização da mesma foram 13 as respostas acertadas por mais de 50% dos doentes (p<0,001). Não foi encontrada relação entre o desempenho do questionário e a duração da doença (p>0,05) e o nível de escolaridade (p>0,05). Conclusão: Através da sessão educativa houve uma melhoria dos conhecimentos dos doentes com glaucoma, relativamente à sua doença.

Prevalence of IgG varicella zoster virus antibodies in the Kuikuro and Kaiabi indigenous communities in Xingu National Park, Brazil, before varicella vaccination

The purpose of the study was to estimate the prevalence of IgG antibodies against varicella zoster virus (VZV) in the two most populated indigenous ethnic groups from Xingu Indigenous National Park, in Brazil, prior to the introduction of vaccination against the disease, and to determine the positive and the negative predictive values of a history of varicella infection. In 2001, 589 inhabitants of two Kuikuro villages and three Kaiabi villages were evaluated and provided information concerning previous varicella infection. An indirect immunosorbent assay (ELISA) to detect IgG anti-VZV antibodies was performed in 224 blood samples - volunteer selection had no interference of anamnesis. IgG prevalence was 80.8% (95% Confidence Interval: 76% - 86%). The seroepidemiology of varicella in Xingu National Park prior to varicella vaccine introduction was comparable to the Brazilian national seroprevalence described in the literature, and so were the positive (98%) and the negative predictive value (41%) of the referred history.

Socio-demographic and epidemiological characteristics associated with human immunodeficiency virus type I (HIV-1) infection in HIV-1-explosed but uninfected individuals, and in HIV-1-infected patients from a southern brasilian population

The ability to control human immunodeficiency virus type 1 (HIV-1) infection and progression of the disease is regulated by host and viral factors. This cross-sectional study describes the socio-demographic and epidemiological characteristics associated with HIV-1 infection in 1,061 subjects attended in Londrina and region, south of Brazil: 136 healthy individuals (Group 1), 147 HIV-1-exposed but uninfected individuals (Group 2), 161 HIV-1-infected asymptomatic patients (Group 3), and 617 patients with AIDS (Group 4). Data were obtained by a standardized questionnaire and serological tests. The age of the individuals ranged from 15.1 to 79.5 years, 54.0% and 56.1% of the Groups 3 and 4 patients, respectively, were men. The major features of groups 2, 3, and 4 were a predominance of education level up to secondary school (55.8%, 60.2% and 62.4%, respectively), sexual route of exposure (88.4%, 87.0% and 82.0%, respectively), heterosexual behavior (91.8%, 75.2% and 83.7%, respectively), and previous sexually transmitted diseases (20.4%, 32.5%, and 38.1%, respectively). The patients with AIDS showed the highest rates of seropositivity for syphilis (25.6%), of anti-HCV (22.3%), and anti-HTLV I/II obtained by two serological screening tests (6.2% and 6.8%, respectively). The results documenting the predominant characteristics for HIV-1 infection among residents of Londrina and region, could be useful for the improvement of current HIV-1 prevention, monitoring and therapeutic programs targeted at this population.

Co-infection between influenza virus and flagellated bacteria

Trypsin is required in the hemagglutinin (HA) cleavage to in vitro influenza viruses activation. This HA cleavage is necessary for virus cell entry by receptor-mediated endocytosis. Bacteria in the respiratory tract are potential sources of proteases that could contribute to the cleavage of influenza virus in vivo. From 47 samples collected from horses, pigs, and from humans, influenza presence was confirmed in 13 and these samples demonstrated co-infection of influenza with flagellated bacteria, Stenotrophomonas maltophilia from the beginning of the experiments. Despite treatment with antibiotics, the bacteria remained resistant in several of the co-infected samples (48.39%). These bacteria, considered opportunistic invaders from environmental sources, are associated with viral infections in upper respiratory tract of hosts. The protease (elastase), secreted by Stenotrophomonas maltophilia plays a role in the potentiation of influenza virus infection. Proteolytic activity was detected by casein agar test. Positive samples from animals and humans had either a potentiated influenza infectivity or cytopathic effect (CPE) in MDCK and NCI H292 cells, Stenotrophomonas maltophilia were always present. Virus and bacteria were observed ultrastructurally. These in vitro findings show that microbial proteases could contribute to respiratory complications by host protease activity increasing inflammation or destroying endogenous cell protease inhibitors.

Relação entre Hipotiroidismo Subclínico e Doença Coronária: um Estudo com Angiografia

INTRODUCTION: The definition of subclinical hypothyroidism (SH) is an asymptomatic state in which free thyroxine (T4) is normal and thyroid-stimulating hormone (TSH) levels are elevated. Its relationship with coronary disease is not clear and has been the subject of recent interest. Current evidence is conflicting and there is a lack of studies supported by coronary angiography. OBJECTIVE: To assess the relationship between SH and the presence and extent of coronary disease diagnosed by angiography. METHODS: We prospectively studied 354 consecutive patients referred for elective coronary angiography. Those with known thyroid disease, documented coronary disease or previous myocardial infarction were excluded. Fasting blood specimens were collected to measure thyroid hormones, lipid profile, high-sensitivity C-reactive protein, fibrinogen and NT-proBNP. Patients with SH were compared with those without to assess differences in clinical characteristics and biochemical and angiographic results. Significant coronary disease was defined as the presence of at least one lesion with > or = 50% luminal stenosis. Lesions with <50% stenosis were considered minimal. RESULTS: SH was diagnosed in 32 (9%) patients. Mean age was similar between the groups. There were more women (66% vs. 39%; p=0.003) and atrial fibrillation was more frequent (25% vs. 11%; p=0.016) in the group of patients with SH. There were no significant differences in the other baseline clinical parameters, and blood biochemistry results were similar in the two groups, with the exception of higher levels of NT-proBNP in SH patients, although without statistical significance. The angiographic results were as follows: significant coronary disease (SH 28.1% vs. non-SH 43.8%; p=0.087); three-vessel disease (9.4% vs. 9.9%; p=0.919); two-vessel disease (12.5% vs. 13.4%; p=0.892); single-vessel disease (6.3% vs. 29.5%; p=0.051); minimal lesions (9.4% vs. 10.9%; p=0.794); and no coronary disease (62.4% vs, 45.3%; p=0.064). CONCLUSION: In this population SH was not associated with the presence or extent of coronary disease diagnosed by coronary angiography.

Doença de Behçet. Experiência de um Serviço de Medicina Interna

Os autores apresentam um estudo retrospectivo dos doentes com Doença de Behçet observados num Serviço de Medicina Interna entre 1982 e 2000. Foram analisados 33 doentes, sendo 20 do sexo masculino e 13 do sexo feminino. A primeira manifestação surgiu, em média, aos 24 anos e a média de idades, na altura do diagnóstico, foi de 32 anos. Todos os doentes apresentaram na sua evolução aftose oral recorrente, surgindo aftose genital em 84,8%, patologia ocular em 81,8%, articular em 75,7%, cutânea em 69,6%, alterações do sistema nervoso central em 27,3%, gerais em 24,2%, vasculares em 21,2%, gastrintestinais em 18,1%, neuropatia periférica em 12,1% e vasculite em 3% dos doentes observados.