Abstract Objectives: Three decades after HIV recognition and its association with AIDS development, many advances have emerged – especially related to prevention and treatment. Undoubtedly, the development of Highly Active Antiretroviral Therapy (HAART) dramatically changed the future of the syndrome that we know today. In the present study, we evaluate the impact of Highly Active Antiretroviral Therapy on macrophage function and its relevance to HIV pathogenesis. Methods: PBMCs were isolated from blood samples and monocytes (CD14+ cells) were purified. Monocyte-Derived Macrophages (MDMs) were activated on classical (MGM-CSF+IFN-γ) or alternative (MIL-4+IL13) patterns using human recombinant cytokines for six days. After this period, Monocyte-Derived Macrophages were stimulated with TLR2/Dectin-1 or TLR4 agonists and we evaluated the influence of HIV-1 infection and Highly Active Antiretroviral Therapy on the release of cytokines/chemokines by macrophages. Results: The data were obtained using Monocyte-Derived Macrophages derived from HIV naïve or from patients on regular Highly Active Antiretroviral Therapy. Classically Monocyte-Derived Macrophages obtained from HIV-1 infected patients on Highly Active Antiretroviral Therapy released higher levels of IL-6 and IL-12 even without PAMPs stimuli when compared to control group. On the other hand, alternative Monocyte-Derived Macrophages derived from HIV-1 infected patients on Highly Active Antiretroviral Therapy released lower levels of IL-6, IL-10, TNF-α, IP-10 and RANTES after LPS stimuli when compared to control group. Furthermore, healthy individuals have a complex network of cytokines/chemokines released by Monocyte-Derived Macrophages after PAMP stimuli, which was deeply affected in MDMs obtained from naïve HIV-1 infected patients and only partially restored in MDMs derived from HIV-1 infected patients even on regular Highly Active Antiretroviral Therapy. Conclusion: Our therapy protocols were not effective in restoring the functional alterations induced by HIV, especially those found on macrophages. These findings indicate that we still need to develop new approaches and improve the current therapy protocols, focusing on the reestablishment of cellular functions and prevention/treatment of opportunistic infections.

OBJETIVO: Descrever o processo de revisão e de integração curricular de um programa de pediatria por meio da criação de uma matriz de competências referenciada nas Diretrizes Curriculares Nacionais. MÉTODOS: Estudo quali-quantitativo de intervenção que avaliou a percepção de estudantes e docentes em relação ao currículo existente (grupos focais e entrevistas semiestruturadas). Discutiram-se os resultados em oficinas de desenvolvimento docente, o que propôs uma matriz baseada em competências para todo o programa de pediatria do 3º ao 6º ano. O novo currículo foi aprovado, implementado e reavaliado após 6 meses. RESULTADOS: Doze estudantes (12%) do 3º ao 6º ano participaram dos grupos focais, e 11 dos 14 professores (78,5%) responderam ao questionário. A maioria referiu falta de integração entre as disciplinas, desconhecimento dos objetivos de aprendizagem dos estágios, poucas oportunidades de práticas e avaliação predominantemente teórica. Nas oficinas de capacitação, foi criada uma matriz curricular integrada por competências após a pactuação entre professores da pediatria e da saúde coletiva. A matriz destacava a competência geral, os objetivos de aprendizagem, oportunidades disponíveis para aprendê-los e o sistema de avaliação. Após 6 meses, 93% (104/112) dos alunos e 79% (11/14) dos professores relataram que percebiam maior integração do programa e destacaram a incorporação da avaliação de desempenho clínico. CONCLUSÃO: A construção coletiva da matriz curricular por competências levou à maior satisfação de docentes e discentes com a nova proposta que, após a implementação, foi percebida como integradora de conteúdos e práticas de ensino da pediatria, tendo qualificado a avaliação de desempenho clínico.

OBJECTIVE: To describe the process of integration and revision of a pediatric program curriculum which resulted in the creation of a competency-based framework recommended in the Brazilian National Curricular Guidelines. METHODS: Quali-quantitative analysis of an intervention evaluating the students and professors' perception of the pediatric program curriculum (focus groups and semi-structured interviews). Results were discussed during teaching development workshops. A competency-based framework was suggested for the pediatric program from the 3rd to the 6th year. The new curriculum was approved, implemented, and reevaluated six months later. RESULTS: Twelve students (12%) from the 3rd to the 6th year participated in the focus groups, and 11 professors (78.5%) answered the questionnaire. Most participants reported lack of integration among the courses, lack of knowledge about the learning goals of the internships, few opportunities of practice, and predominance of theoretical evaluation. In the training workshops, a competency-based curriculum was created after pediatrics and collective health professors reached an agreement. The new curriculum was focused on general competency, learning goals, opportunities available to learn these goals, and evaluation system. After six months, 93% (104/112) of students and 79% (11/14) of professors reported greater integration of the program and highlighted the inclusion of the clinical performance evaluation. CONCLUSION: The collective creation of a competency-based curriculum promoted higher satisfaction of students and professors. After being implemented, the new curriculum was considered to integrate the teaching practices and contents, improving the quality of the clinical performance evaluation.

OBJETIVO: Padronizar reação em cadeia da polimerase para diagnóstico de tuberculose pulmonar, comparando os resultados obtidos com as técnicas microbiológicas clássicas, e analisar seu uso numa região de alta prevalência da tuberculose. MÉTODOS: Foram descontaminadas, após a baciloscopia, 42 amostras de escarro de pacientes. Em seguida, procedeu-se ao cultivo em Lowenstein-Jensen e à reação em cadeia da polimerase com "primers" que amplificam um fragmento de 123 pares de base do genoma do Mycobacterium tuberculosis. RESULTADOS: Das 42 amostras de escarro, 10 apresentaram cultura positiva para M. tuberculosis. Dez foram positivas à baciloscopia e 16 mostraram-se positivas na reação em cadeia da polimerase. A sensibilidade e especificidade do teste em relação à cultura foi de 90% e 81%, respectivamente. CONCLUSÕES: A reação em cadeia da polimerase tem sensibilidade comparável à da cultura e pode ser realizada em apenas um dia, resultando em tratamento precoce e melhor controle da doença. A padronização e avaliação de técnicas de biologia molecular no diagnóstico da tuberculose no Brasil é imprescindível na discussão da implantação deste exame na rotina diagnóstica em centros de referência.

INTRODUCTION: The recent increase in the number of tuberculosis cases has called the world's attention once again to a perennial health problem, especially prevalent in developing countries. The time elapsed between the diagnosis and the institution of therapy is an obstacle to tuberculosis control and there is an urgent need for the development of techniques for the disease's rapid diagnosis. To achieve this goal, molecular biology techniques have been exhaustively investigated. This work describes the use of a polymerase chain reaction for rapid diagnosis of tuberculosis in a developing country. The sensitivity and specificity of this technique is compared to standard techniques used in the microbiology laboratory. METHODS: This study was undertaken in Ribeirão Preto, S. Paulo State, Brazil. Forty-two sputum samples from suspected cases of tuberculosis attending the municipal health care centers were sent to the microbiology laboratory. The samples were processed for the detection of Mycobacterium tuberculosis by acid-fast bacilli determination, culture in Lowenstein-Jensen medium, and by a polymerase chain reaction that amplified a fragment of 123 base pairs of the Mycobacterium tuberculosis genome. RESULTS: Of the forty-two samples studied, one was contaminated and excluded from the study, ten were culture positive, ten were positive for the presence of acid-fast bacilli, and sixteen were polymerase chain reaction positive. The sensitivity and specificity of this technique were 90% and 81%, respectively. CONCLUSIONS: The polymerase chain reaction presented a sensitivity comparable to the culture and the whole procedure took only one day to complete. The results presented here make it a strong candidate for rapid diagnosis of tuberculosis in clinical settings making it possible to begin the specific therapy early in the course of the disease. However, standardization of the technique is necessary, and the correlation with clinical findings is of paramount importance due to the high sensitivity of this technique.

OBJETIVO: Padronizar reação em cadeia da polimerase para diagnóstico de tuberculose pulmonar, comparando os resultados obtidos com as técnicas microbiológicas clássicas, e analisar seu uso numa região de alta prevalência da tuberculose. MÉTODOS: Foram descontaminadas, após a baciloscopia, 42 amostras de escarro de pacientes. Em seguida, procedeu-se ao cultivo em Lowenstein-Jensen e à reação em cadeia da polimerase com "primers" que amplificam um fragmento de 123 pares de base do genoma do Mycobacterium tuberculosis. RESULTADOS: Das 42 amostras de escarro, 10 apresentaram cultura positiva para M. tuberculosis. Dez foram positivas à baciloscopia e 16 mostraram-se positivas na reação em cadeia da polimerase. A sensibilidade e especificidade do teste em relação à cultura foi de 90% e 81%, respectivamente. CONCLUSÕES: A reação em cadeia da polimerase tem sensibilidade comparável à da cultura e pode ser realizada em apenas um dia, resultando em tratamento precoce e melhor controle da doença. A padronização e avaliação de técnicas de biologia molecular no diagnóstico da tuberculose no Brasil é imprescindível na discussão da implantação deste exame na rotina diagnóstica em centros de referência.

INTRODUCTION: The recent increase in the number of tuberculosis cases has called the world's attention once again to a perennial health problem, especially prevalent in developing countries. The time elapsed between the diagnosis and the institution of therapy is an obstacle to tuberculosis control and there is an urgent need for the development of techniques for the disease's rapid diagnosis. To achieve this goal, molecular biology techniques have been exhaustively investigated. This work describes the use of a polymerase chain reaction for rapid diagnosis of tuberculosis in a developing country. The sensitivity and specificity of this technique is compared to standard techniques used in the microbiology laboratory. METHODS: This study was undertaken in Ribeirão Preto, S. Paulo State, Brazil. Forty-two sputum samples from suspected cases of tuberculosis attending the municipal health care centers were sent to the microbiology laboratory. The samples were processed for the detection of Mycobacterium tuberculosis by acid-fast bacilli determination, culture in Lowenstein-Jensen medium, and by a polymerase chain reaction that amplified a fragment of 123 base pairs of the Mycobacterium tuberculosis genome. RESULTS: Of the forty-two samples studied, one was contaminated and excluded from the study, ten were culture positive, ten were positive for the presence of acid-fast bacilli, and sixteen were polymerase chain reaction positive. The sensitivity and specificity of this technique were 90% and 81%, respectively. CONCLUSIONS: The polymerase chain reaction presented a sensitivity comparable to the culture and the whole procedure took only one day to complete. The results presented here make it a strong candidate for rapid diagnosis of tuberculosis in clinical settings making it possible to begin the specific therapy early in the course of the disease. However, standardization of the technique is necessary, and the correlation with clinical findings is of paramount importance due to the high sensitivity of this technique.