Magnetic purification of biotinylated cDNA removes false priming and ensures strand-specificity of RT-PCR for enteroviral RNAs

The detection of replicative intermediate RNAs as markers of active replication of RNA viruses is an essential tool to investigate pathogenesis in acute viral infections, as well as in their long-term sequelae. In this regard, strand-specific PCR has been used widely to distinguish (-) and (+) enteroviral RNAs in pathogenesis studies of diseases such as dilated cardiomyopathy. It has been generally assumed that oligonucleotide-primed reverse transcription of a given RNA generates only the corresponding specific cDNA, thus assuring the specificity of a PCR product amplified from it. Nevertheless, such assumed strand-specificity is a fallacy, because falsely primed cDNAs can be produced by RNA reverse transcription in the absence of exogenously added primers, (cDNA(primer)(-)), and such falsely primed cDNAs are amplifiable by PCR in the same way as the correctly primed cDNAs. Using as a prototype the coxsackievirus B5 (CVB5), a (+) strand RNA virus, it was shown that cDNA(primer)(-) renders the differential detection of viral (-) and (+) RNAs by conventional PCR virtually impossible, due to gross non-specificity. Using in vitro transcribed CVB5 RNAs (+) and (-), it was shown that cDNA(primer)(-) could be removed effectively by magnetic physical separation of correctly primed biotinylated cDNA. Such strategy enabled truly strand-specific detection of RNA (-) and (+), not only for CVB5, but also for other non-polio enteroviruses. These findings indicate that previous conclusions supporting a role for the persistence of actively replicating enterovirus in the pathogenesis of chronic myocarditis should be regarded with strong skepticism and purification of correctly primed cDNA should be used for strand-specific PCR of viral RNA in order to obtain reliable information on this important subject. (C) 2009 Elsevier B.V. All rights reserved.

Changing attitudes, beliefs and feelings towards food in bulimic patients

Eating attitudes are defined as beliefs. thoughts, feelings and behaviors towards food. Bulimia nervosa (BN) is ail eating disorder, in which the eating, attitudes are Seriously disturbed. Studies that evaluated nutritional aspects of BN focus mainly oil food intake, dietary restriction and binge eating. while the follow-up Studies evaluate mainly clinical symptoms. The objective of this study was to evaluate eating attitudes of patients with BN. during and after cognitive-behavioral intervention. Thirty nine (39) BN female patients received cognitive behavioral treatment with a Multidisciplinary team and had eating attitudes assessed by a questionnaire developed for this research. Frequencies of the attitudes assessed were compared at baseline. after 12 weeks and 24 weeks of treatment. After treatment, patients had less distorted beliefs about food, less guilty after eating ""forbidden"" foods and they felt more tranquil while caring outside home. Other negative behaviors, as dietary restriction, the desire of not cat, being angry when feeling hungry and using the food to relive stress. persisted. Eating attitudes of patients with BN are hard to be changed in a short-term. More attention to this disease`s component and new approaches to treatment are needed in order to have a better recovery.

False-positive results of a rapid K39-based strip test and Chagas disease

Background: The definitive diagnosis of visceral. leishmaniasis (VL) requires invasive procedures with demonstration of amastigotes in tissue or promastigotes in culture. Unfortunately, these approaches require laboratory materials not available in poor countries where the disease is endemic. The correct diagnosis of VL is important, and made more difficult by the fact that several common tropical diseases such as malaria, disseminated tuberculosis, and enteric fever share the same clinical presentation. Serological tests have been developed to replace parasitological diagnosis in the field. A commercially available K39-based strip test for VL has been developed for this purpose. The endemic area of leishmaniasis in Brazil overlaps the endemic area of Chagas disease, a disease that can cause false-positive serological test results. The aim of this study was to evaluate the incidence of false-positive exams using a rapid test for VL in patients with Chagas disease. Methods: A rapid test based on the recombinant K39 antigen of Leishmania was used in: (1) 30 patients with confirmed Chagas disease, (2) 30 patients with a serological diagnosis of Chagas disease by ELISA, indirect immunofluorescence, indirect hemagglutination, and chemiluminescence, (3) 30 healthy patients from a non-endemic area as the control group, (4) 30 patients with confirmed VL, and (5) 20 patients with proved cutaneous leishmaniasis. Results: The sensitivity and specificity of the rapid strip test were 100% when compared with healthy volunteers and those with confirmed Chagas disease. One false-positive result occurred in the group with Chagas disease diagnosed by serological tests (specificity of 96%). Conclusion: The rapid test based on recombinant K39 is a useful diagnostic assay, and a false-positive result rarely occurs in patients with a serological diagnosis of Chagas disease. (C) 2008 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Uncovering false positives on a virtual screening search for cruzain inhibitors

Some unexpected promiscuous inhibitors were observed in a virtual screening protocol applied to select cruzain inhibitors from the ZINC database. Physical-chemical and pharmacophore model filters were used to reduce the database size. The selected compounds were docked into the cruzain active site. Six hit compounds were tested as inhibitors. Although the compounds were designed to be nucleophilically attacked by the catalytic cysteine of cruzain, three of them showed typical promiscuous behavior, revealing that false positives are a prevalent concern in VS programs. (C) 2007 Elsevier Ltd. All rights reserved.

Bleaching of melanin in the epidermis of South American fur seal and its application on enzyme immunohistochemistry

The South American fur seal (Arctocephalus australis) is an amphibious marine mammal distributed along the Atlantic and Pacific coasts of South America. The species is well adjusted to different habitats due to the morphology of its fin-like members and due to some adaptations in their integumentary system. Immunohistochemical studies are very important to evaluate the mechanisms of skin adaptation due the differential expression of the antigens present in the tissue depending of the region of the body surface. However, its strongly pigmented (melanin) epidermis prevents the visualization of the immuno-histochemical chromogens markers. In this study a melanin bleaching method was developed aimed to allow the visualization of the chromogens without interfering in the antigen-antibody affinity for immunohistochemistry. The analysis of PCNA (proliferating cell nuclear antigen) index in the epidermis of A. australis by immunohistochemistry with diaminobenzidine (DAB) as chromogen was used to test the method. The bleaching of the melanin allowed to obtain the cell proliferation index in epidermis and to avoid false positive results without affecting the immunohistochemical results.

Method for determination of root curvature radius using cone-beam computed tomography images

This article describes and discusses a method to determine root curvature radius by using cone-beam computed tomography (CBCT). The severity of root canal curvature is essential to select instrument and instrumentation technique. The diagnosis and planning of root canal treatment have traditionally been made based on periapical radiography. However, the higher accuracy of CBCT images to identify anatomic and pathologic alterations compared to panoramic and periapical radiographs has been shown to reduce the incidence of false-negative results. In high-resolution images, the measurement of root curvature radius can be obtained by circumcenter. Based on 3 mathematical points determined with the working tools of Planimp® software, it is possible to calculate root curvature radius in both apical and coronal directions. The CBCT-aided method for determination of root curvature radius presented in this article is easy to perform, reproducible and allows a more reliable and predictable endodontic planning, which reflects directly on a more efficacious preparation of curved root canals.

Fine needle aspiration biopsy in the oral cavity and head and neck region

The objective of the current study was to evaluate the sensitivity, specificity and accuracy of fine needle aspiration biopsy (FNAB) of submucous nodules from the oral cavity and head and neck region as an auxiliary diagnostic tool. Fifty patients with nodule lesions in the oral cavity and the head and neck region were selected. All of them were submitted to FNAB and to either incisional or excisional biopsy. The diagnoses from the FNABs were compared with the biopsy diagnosis as the gold standard. All the cases of FNAB were analyzed by a single oral pathologist prior to the biopsy diagnosis. The results showed that the sensitivity of FNAB was 75%, its specificity was 96% and its accuracy was 58.8%. The false positive and false negative rates were 6.7% and 13.3%, respectively. The positive predictive value was 86% and the negative predictive value was 93%. The inconclusive rate was 16/50. FNAB displayed a high success rate for identifying both malignant and benign lesions, but a low accuracy for making a final diagnosis.

Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)

Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity.

Laços mal-atados como efeito de funcionamento falso-self em tempos de desconfiança

A contemporaneidade contempla formas de estruturação da subjetividade amplamente perpassadas por condições de existência socioculturais e econômicas. Estas, por sua vez, têm promovido laços intersubjetivos frágeis, cujo funcionamento psíquico pode ter uma organização de base "falso-self". Autores também demonstram que diversas instituições atuais se tornaram "fluidas" na passagem da modernidade para a "modernidade líquida". Isto contribuiu para a perda de referências sólidas que, outrora, davam sentido e direcionamento para o ser humano. Em decorrência há uma interferência direta na construção da "confiança madura", visto que os sinais de angústia da contemporaneidade podem ser um desorganizador do funcionamento psíquico, além de promoverem o isolamento, a desconfiança no ambiente e no outro, laços mal-atados e funcionamentos de base "falso-self".

Desenvolvimento da identidade em adolescentes estudantes do ensino médio

Identidade é composta pelos valores, crenças e metas com que um indivíduo está comprometido. Existem quatro estados de identidade: difusão, pré-fechamento, moratória e identidade estabelecida, identificados de acordo com a presença de exploração das alternativas e com a intensidade dos compromissos. Este trabalho verificou os estados de identidade em que se encontravam 753 adolescentes de 15 a 18 anos, estudantes do Ensino Médio de escolas públicas de São Paulo que responderam ao EOMEIS 2, uma escala tipo Likert que avalia os estados de identidade. A maioria dos adolescentes encontrava-se em moratória. Verificou-se uma associação positiva entre o sexo feminino e os estados de moratória e identidade estabelecida; entre a primeira série do Ensino Médio e o estado de pré-fechamento.

Construção e validação da Escala de Crenças Parentais e Práticas de Cuidado (E-CPPC) na primeira infância

O presente estudo teve como objetivo construir e validar uma escala de crenças parentais e práticas de cuidado na primeira infância para o contexto brasileiro. Após a construção teórica dos itens, a versão final da escala foi aplicada em dois estudos com populações distintas (estudo preliminar - 250 mães; estudo final - 600 mães), visando avaliar a frequência de comportamento e o grau de importância atribuído às práticas. Para as análises psicométricas, foram realizadas: análise fatorial e o cálculo da consistência interna. Em ambos os estudos a análise fatorial resultou em dois fatores, sendo o primeiro denominado "cuidados primários" e o segundo "estimulação". Os alfas de Cronbach apresentaram-se satisfatórios, variando de 0,68 a 0,83. Os resultados do estudo demonstraram a validade e precisão da escala, a qual poderá ser utilizada em futuras pesquisas, bem como em práticas de intervenção familiar, principalmente durante os primeiros anos de vida das crianças.

As disciplinas pedagógicas na formação e na construção de representações sobre o trabalho docente: visões de alunos de licenciatura em Química e Física

Este estudo tem como objetivo analisar as concepções de licenciandos em Química e em Física sobre a prática profissional e as marcas deixadas pelas disciplinas pedagógicas na sua formação, assim como identificar sinais de mudanças em suas concepções durante a realização do curso de Licenciatura. Para tanto, textos foram produzidos por 54 alunos matriculados em cursos de Licenciatura de duas universidades públicas do estado de São Paulo, nos quais os licenciandos expressaram suas experiências de formação e apontaram elementos que consideram que terão alguma influência em sua atuação profissional. Os textos revelaram que as ideias que os licenciandos carregam são consistentes com muitas das visões e objetivos apresentados em cursos de formação de professores. Nas produções dos estudantes faz-se presente o desejo de compartilhar e promover em seus futuros alunos o interesse pela Química e a Física. Os seus comentários refletiram visões de professores que tiveram no passado, os quais gostariam de imitar, e de outros que não gostariam de ser. Os comentários também tornaram evidente a vontade dos licenciandos em se estabelecerem como professores que se preocupam com seus alunos. Os textos foram analisados segundo a Análise do Discurso de linha francesa, como tem sido divulgada por Eni Orlandi, especialmente a noção de formação discursiva.

Diferenças na adesão ao tratamento da tuberculose em relação ao sexo no distrito de saúde da Freguesia do Ó/Brasilândia - São Paulo

Os motivos para as diferenças epidemiológicas e para a adesão ao tratamento da tuberculose em relação a homens e mulheres são desconhecidos. Este trabalho tem como objetivo verificar diferenças na adesão ao tratamento da tuberculose em relação ao sexo; identificar aspectos facilitadores e dificultadores para a adesão ao tratamento da tuberculose em relação ao sexo; analisar as crenças consideradas importantes para a adesão ao tratamento da tuberculose. Foi utilizado o referencial teórico do Modelo de Crenças em Saúde de Rosenstock e a técnica da Análise de Conteúdos de Bardin. Foram realizadas 28 entrevistas semiestruturadas com homens e mulheres em tratamento supervisionado de tuberculose do Distrito de Saúde da Freguesia do Ó/Brasilândia. Os resultados mostraram que o perfil daqueles que falharam na terapêutica da tuberculose em relação ao sexo foi: mulher - solteira e separada, com atividade remunerada não comprovada, nível de escolaridade entre fundamental I completo e ensino médio completo; homem - casado, com atividade remunerada comprovada, nível de escolaridade entre ensino fundamental II completo e ensino médio completo. Os aspectos facilitadores encontrados para a boa adesão residem no bom atendimento dos profissionais de saúde e na percepção, por parte do paciente, da sua melhora de saúde. As crenças para a boa adesão ao tratamento no sexo masculino e feminino foram: bom atendimento do serviço de saúde e bom tratamento (em relação aos medicamentos).

Investigation of measles IgM-seropositive cases of febrile rash illnesses in the absence of documented measles virus transmission, State of São Paulo, Brazil, 2000-2004

Introdução: Revisar os casos de doenças febris exantemáticas com IgM reagente contra o sarampo, no estado de São Paulo, Brasil, durante os cinco anos seguidos a interrupção da transmissão do vírus do sarampo. Métodos: Nós revisamos 463 casos de doenças febris exantemáticas com IgM reagente contra o sarampo, no estado de São Paulo, Brasil, de 2000 a 2004. Indivíduos vacinados contra o sarampo 56 dias antes da coleta de amostra foram considerados expostos à vacina. Soros da fase aguda e de convalescença foram testados para a evidência de infecção de sarampo, rubéola, parvovírus B19 e herpes vírus 6. Na ausência de soroconversão para imunoglobulina G contra o sarampo, casos com IgM reagente contra o sarampo foram considerados falsos positivos em pessoas com evidência de outras infecções virais. Resultados: Entre as 463 pessoas com doenças febris exantemáticas que testaram positivo para anticorpos IgM contra o sarampo durante o período, 297 (64 por cento) pessoas foram classificadas como expostas à vacina. Entre os 166 casos não expostos à vacina, 109 (66 por cento) foram considerados falsos positivos baseado na ausência de soroconversão, dos quais 21 (13 por cento) tiveram evidência de infecção por vírus da rubéola, 49 (30 por cento) parvovírus B19 e 28 (17 por cento) infecção por herpes vírus humano 6. Conclusões: Após a interrupção da transmissão do vírus do sarampo é necessária exaustiva investigação dos casos com IgM reagente contra o sarampo, especialmente dos casos não expostos à vacina. Testes laboratoriais para etiologias das doenças febris exantemáticas ajudam na interpretação destes casos

Psychometric evaluation of the Disordered Eating Attitude Scale (DEAS)

Eating attitudes are defined as beliefs, thoughts, feelings, behaviors and relationship with food. They could influence people’s food choices and their health status. Objective: This study aimed to adapt from Portuguese to English the Disordered Eating Attitude Scale (DEAS) and evaluate its validity and reliability. The original scale in Portuguese was translated and adapted into English and was applied to female university students of University of Minnesota—USA (n = 224). Internal consistency was determined (Cronbach’s Alpha). Convergent validity was assessed by correlations between Eating Attitude Test-26 (EAT-26) and Restrain Scale (RS). Reliability was evaluated applying twice the scale to a sub-sample (n = 30). The scale was back translated into Portuguese and compared with the original version and discrepancies were not found. The internal consistency was .76. The DEAS total score was significantly associated with EAT-26 (r = 0.65) and RS (r = 0.69) scores. The correlation between test–retest was r = 0.9. The English version of DEAS showed appropriate internal consistency, convergent validity and test–retest reliability and will be useful to assess eating attitudes in different population groups in English spoken countries