Electrophoretic characterization of LNP/AAV-encapsulated nucleic acids: Strengths and weaknesses.

The use of nucleic acids (NAs) has revolutionized medical approaches and ushered in a new era of combating various diseases. Accordingly, there is an increasing demand for accurate identification, localization, quantification, and characterization of NAs encapsulated in nonviral or viral vectors. The vast spectrum of molecular dimensions and intra- and intermolecular interactions presents a formidable obstacle for NA analytical development. Typically, the comprehensive analysis of encapsulated NAs, free NAs, and their spatial distribution poses a challenge that is seldom tackled in its complete complexity. The identification of appropriate physicochemical methodologies for large nonencapsulated or encapsulated NAs is particularly intricate and necessitates an evaluation of the analytical outcomes and their appropriateness in addressing critical quality attributes. In this work, we examine the analytics of non-encapsulated or encapsulated large NAs (>500 nucleotides) utilizing capillary electrophoresis (CE) and liquid chromatography (LC) methodologies such as free zone CE, gel CE, affinity CE, and ion pair high-performance liquid chromatography (HPLC). These methodologies create a complete picture of the NA's critical quality attributes, including quantity, identity, purity, and content ratio.

Evaluation of comprehensive care for older adults in primary care services.

OBJECTIVE: To evaluate the performance of comprehensive care for older adults in primary care services in the Brazilian Unified Health System in the state of São Paulo, Brazil. METHODS: A total of 157 primary care services from five health regions in midwestern São Paulo responded, from October to December 2014, the pre-validated 2014 questionnaire for primary care services assessment and monitoring. We selected 155 questions, based on national policies and guidelines on this theme. The responses indicate the service performance in older adults' care, clustered into three areas of analysis: health care for active and healthy aging (45 indicators, d1), chronic noncommunicable diseases care (89 indicators, d2), and support network in aging care (21 indicators, d3). Performance was measured by the sum of positive (value 1) or negative (value 0) responses for each indicator. Services were clustered according to k-means of the performance scores of each domain. After weighting the domains (Z tests), we estimated the associations between the scores of each domain and independent management variables (typology, planning and evaluation of services), with simple and multiple linear regression. RESULTS: Chronic noncommunicable diseases care (d2) showed, for all clusters, better average performance (55.7) than domains d1 (35.4) and d3 (39.2). Service performance in the general area of planning and evaluation associates with the performance of older adults' care. CONCLUSIONS: The evaluated services had incipient implementation of comprehensive care for older adults. The evaluation framework can contribute to processes to improve the quality of primary health care.

Evaluation of comprehensive care for older adults in primary care services / Avaliação da atenção integral ao idoso em serviços de atenção primária

ABSTRACT OBJECTIVE To evaluate the performance of comprehensive care for older adults in primary care services in the Brazilian Unified Health System in the state of São Paulo, Brazil. METHODS A total of 157 primary care services from five health regions in midwestern São Paulo responded, from October to December 2014, the pre-validated 2014 questionnaire for primary care services assessment and monitoring. We selected 155 questions, based on national policies and guidelines on this theme. The responses indicate the service performance in older adults' care, clustered into three areas of analysis: health care for active and healthy aging (45 indicators, d1), chronic noncommunicable diseases care (89 indicators, d2), and support network in aging care (21 indicators, d3). Performance was measured by the sum of positive (value 1) or negative (value 0) responses for each indicator. Services were clustered according to k-means of the performance scores of each domain. After weighting the domains (Z tests), we estimated the associations between the scores of each domain and independent management variables (typology, planning and evaluation of services), with simple and multiple linear regression. RESULTS Chronic noncommunicable diseases care (d2) showed, for all clusters, better average performance (55.7) than domains d1 (35.4) and d3 (39.2). Service performance in the general area of planning and evaluation associates with the performance of older adults' care. CONCLUSIONS The evaluated services had incipient implementation of comprehensive care for older adults. The evaluation framework can contribute to processes to improve the quality of primary health care.

RESUMO OBJETIVO Avaliar o desempenho da atenção integral ao idoso em serviços de atenção primária do Sistema Único de Saúde no estado de São Paulo, Brasi lMÉTODOS Um total de 157 serviços de atenção primária de cinco regiões de saúde do centro-oeste paulista respondeu, de outubro a dezembro de 2014, o instrumento pré-validado Questionário de Avaliação e Monitoramento de Serviços de Atenção Básica 2014. Foram selecionadas 155 questões, com base nas políticas e diretrizes nacionais sobre essa temática. As respostas indicam o desempenho do serviço na atenção ao idoso, agrupadas em três domínios de análise: atenção à saúde para o envelhecimento ativo e saudável (45 indicadores, d1), atenção às doenças crônicas não transmissíveis (89 indicadores, d2) e rede de apoio na atenção ao envelhecimento (21 indicadores, d3). A medida de desempenho foi a soma de respostas positivas (valor 1) ou negativas (valor 0) para cada indicador. Os serviços foram agrupados segundo k-médias dos escores de desempenho de cada um dos domínios. Após a ponderação dos domínios (testes Z), foram estimadas as associações entre os escores de cada domínio e variáveis independentes de gestão (tipologia, planejamento e avaliação dos serviços), por meio de regressão linear simples e múltipla. RESULTADOS A atenção às doenças crônicas não transmissíveis (d2) mostrou, para todos os agrupamentos, melhor desempenho médio (55,7) do que os domínios d1 (35,4) e d3 (39,2). O desempenho do serviço na área geral de planejamento e avaliação esteve associado ao desempenho da atenção ao idoso. CONCLUSÕES Os serviços avaliados apresentaram implementação incipiente da atenção integral ao idoso. O quadro avaliativo pode contribuir para processos de melhoria da qualidade da atenção primária à saúde.

Hair Cortisol Measurement by an Automated Method.

We present the development of the first procedure for hair cortisol measurement through an automated method. Hair samples were obtained from 286 individuals. After cortisol extraction, samples were measured in a Siemens Immulite 2000 (Gwynedd, UK) automated chemoluminiscent immunoassay analyzer. Normal reference values were obtained from hair cortisol levels measured in 213 healthy individuals with low levels of stress. Hair cortisol concentration median was 55 pg/mg hair (2.5-97.5 percentile (40-128)) in healthy individuals with low levels of stress and 250 pg/mg hair (range 182-520) in stressed individuals. No significant differences were observed in hair cortisol levels between subjects with and without dye (40 (40-107) and 40 (40-155) pg/mg hair, respectively; p = 0.128). The novel procedure presented here shows an adequate analytical performance.

Importância do gerenciamento local para uma atenção primária à saúde nos moldes de Alma-Ata

[RESUMO]. Objetivo. Descrever as características da gerência das unidades de atenção primária à saúde e o perfil dos gerentes e discutir as implicações desses elementos para a efetivação dos pressupostos do Sistema Único de Saúde no Brasil de forma coerente com as proposições de Alma-Ata. Métodos. Estudo descritivo, transversal, com dados colhidos pelo questionário de Avaliação da Qualidade de Serviços de Atenção Básica (QualiAB), um instrumento autoaplicado via web. O QualiAB foi respondido voluntariamente por 157 gerentes de unidades básicas de saúde de 41 municípios do estado de São Paulo entre outubro e dezembro de 2014. Resultados. Das 157 unidades, 67 (42,7%) eram unidades de saúde da família e 58 (36,9%) eram unidades básicas de saúde de organização “tradicional”; 95 (60,5%) se localizavam em região urbana periférica. No momento do estudo, oito (5,0%) unidades não possuíam gerente e oito (5,0%) eram gerenciadas por secretários municipais de saúde. Quase 80% dos gerentes eram enfermeiros e desempenhavam múltiplas funções além da gerência. O matriciamento (supervisão técnica como forma de educação permanente) era feito em 75 (47,7%) unidades; 60 (38,2%) unidades não contavam com nenhum tipo de matriciamento. A participação em processos avaliativos foi referida por 130 (82,8%) serviços. Os principais desdobramentos induzidos por avaliações foram planejamento e reprogramação das atividades com participação da equipe multiprofissional em 40 unidades (25,5%) e definição de um plano anual de atividades em 38 (24,2%). Não tiveram acesso aos resultados das avaliações 29 unidades (17,8%). Conclusão. O estudo recoloca a importância da gestão do trabalho e a necessidade de (re) investir na formação e valorização do gerenciamento local como estratégia para efetivar uma atenção primária à saúde capaz de promover a saúde como direito e condição de cidadania.

[ABSTRACT]. Objective. To describe the characteristics of the management process in primary health care units as well as the profile of managers, and to discuss the implications of these elements for the attainment of Unified Health System principles in Brazil in accordance with the propositions of the Alma-Ata Declaration. Method. This descriptive, cross-sectional study used data collected with the Primary Care Service Quality Assessment tool (QualiAB), a self-administered, web-based instrument. QualiAB was voluntarily answered by 157 unit managers from 41 municipalities in the state of São Paulo from October to December 2014. Results. Of 157 units, 67 (42.7%) were family health care units and 58 (36.9%) were “traditional” units; 95 (60.5%) were located in urban peripheries. At the time of the study, eight (5.0%) units did not have a manager and eight (5.0%) were managed by the city health secretary. Almost 80% of the managers were nurses and performed multiple tasks in addition to management. Multidisciplinary support (technical supervision as a means of continuing education) was available in 75 (47.7%) units; 60 (38.2%) units did not have any kind of multidisciplinary support. Participation in evaluative processes was mentioned in 130 (82.8%) units. The main results of evaluations were planning and reprogramming of activities with the engagement of the multiprofessional team in 40 units (25.5%) and definition of an annual activity plan in 38 (24.2%). Twenty-nine units (17.8%) did not have access to the results of evaluations. Conclusion. The study supports the importance of work process management and the need to (re)invest in training and recognition of local management as a strategy to produce primary health care that is capable of promoting health as a right and a condition of citizenship.

[RESUMEN]. Objetivo. Describir las características de la gestión de las unidades de atención primaria de salud y el perfil de los gerentes, y analizar las implicaciones de esos elementos en la puesta en práctica de los principios del Sistema Único de Salud de Brasil de forma coherente con las propuestas de Alma- Ata. Métodos. Estudio descriptivo, transversal, con datos recolectados a través del cuestionario de Evaluación de la Calidad de Servicios de Atención Básica (QualiAB), un instrumento autoadministrado a través de Internet. En total 157 gerentes de Unidades Básicas de Salud de 41 municipios del estado de São Paulo respondieron voluntariamente el QualiAB entre octubre y diciembre de 2014. Resultados. De las 157 unidades, 67 (42,7%) eran unidades salud de la familia y 58 (36,9%) eran unidades básicas de salud de organización “tradicional”; 95 (60,5%) están ubicadas en una región urbana periférica. En el momento del estudio, ocho (5,0%) unidades no poseían gerente y ocho (5,0%) eran gestionadas por secretarios municipales de salud. Casi el 80% de los gerentes eran enfermeros y desempeñaban múltiples funciones además de la gerencia. En 75 (47,7%) unidades se disponía de apoyo multidisciplinario (supervisión técnica como forma de educación permanente); 60 (38,2%) unidades no contaban con ningún tipo de apoyo multidisciplinario. La participación en procesos de evaluación fue referida por 130 (82,8%) servicios. Las principales modificaciones inducidas por las evaluaciones fueron la planificación y reprogramación de las actividades con participación del equipo multiprofesional en 40 unidades (25,5%) y la definición de un plan anual de actividades en 38 (24,2%). No tuvieron acceso a los resultados de las evaluaciones 29 unidades (17,8%). Conclusión. El estudio subraya la importancia de la gestión del trabajo y la necesidad de reinvertir en la formación y valorización de la gestión local como estrategia para hacer efectiva una atención primaria de salud capaz de promover la salud como derecho y condición de ciudadanía.

Steady-state analysis of enzymes with non-Michaelis-Menten kinetics: The transport mechanism of Na+/K+-ATPase.

Procedures to define kinetic mechanisms from catalytic activity measurements that obey the Michaelis-Menten equation are well established. In contrast, analytical tools for enzymes displaying non-Michaelis-Menten kinetics are underdeveloped, and transient-state measurements, when feasible, are therefore preferred in kinetic studies. Of note, transient-state determinations evaluate only partial reactions, and these might not participate in the reaction cycle. Here, we provide a general procedure to characterize kinetic mechanisms from steady-state determinations. We described non-Michaelis-Menten kinetics with equations containing parameters equivalent to kcat and Km and modeled the underlying mechanism by an approach similar to that used under Michaelis-Menten kinetics. The procedure enabled us to evaluate whether Na+/K+-ATPase uses the same sites to alternatively transport Na+ and K+ This ping-pong mechanism is supported by transient-state studies but contradicted to date by steady-state analyses claiming that the release of one cationic species as product requires the binding of the other (ternary-complex mechanism). To derive robust conclusions about the Na+/K+-ATPase transport mechanism, we did not rely on ATPase activity measurements alone. During the catalytic cycle, the transported cations become transitorily occluded (i.e. trapped within the enzyme). We employed radioactive isotopes to quantify occluded cations under steady-state conditions. We replaced K+ with Rb+ because 42K+ has a short half-life, and previous studies showed that K+- and Rb+-occluded reaction intermediates are similar. We derived conclusions regarding the rate of Rb+ deocclusion that were verified by direct measurements. Our results validated the ping-pong mechanism and proved that Rb+ deocclusion is accelerated when Na+ binds to an allosteric, nonspecific site, leading to a 2-fold increase in ATPase activity.

[Importance of local management for delivery of primary health care according to Alma-Ata principlesImportancia de la gestión local para una atención primaria de salud según las propuestas de Alma-Ata]. / Importância do gerenciamento local para uma atenção primária à saúde nos moldes de Alma-Ata.

OBJECTIVE: To describe the characteristics of the management process in primary health care units as well as the profile of managers, and to discuss the implications of these elements for the attainment of Unified Health System principles in Brazil in accordance with the propositions of the Alma-Ata Declaration. METHOD: This descriptive, cross-sectional study used data collected with the Primary Care Service Quality Assessment tool (QualiAB), a self-administered, web-based instrument. QualiAB was voluntarily answered by 157 unit managers from 41 municipalities in the state of São Paulo from October to December 2014. RESULTS: Of 157 units, 67 (42.7%) were family health care units and 58 (36.9%) were "traditional" units; 95 (60.5%) were located in urban peripheries. At the time of the study, eight (5.0%) units did not have a manager and eight (5.0%) were managed by the city health secretary. Almost 80% of the managers were nurses and performed multiple tasks in addition to management. Multidisciplinary support (technical supervision as a means of continuing education) was available in 75 (47.7%) units; 60 (38.2%) units did not have any kind of multidisciplinary support. Participation in evaluative processes was mentioned in 130 (82.8%) units. The main results of evaluations were planning and reprogramming of activities with the engagement of the multiprofessional team in 40 units (25.5%) and definition of an annual activity plan in 38 (24.2%). Twenty-nine units (17.8%) did not have access to the results of evaluations. CONCLUSION: The study supports the importance of work process management and the need to (re)invest in training and recognition of local management as a strategy to produce primary health care that is capable of promoting health as a right and a condition of citizenship.

OBJETIVO: Describir las características de la gestión de las unidades de atención primaria de salud y el perfil de los gerentes, y analizar las implicaciones de esos elementos en la puesta en práctica de los principios del Sistema Único de Salud de Brasil de forma coherente con las propuestas de Alma- Ata. MÉTODOS: Estudio descriptivo, transversal, con datos recolectados a través del cuestionario de Evaluación de la Calidad de Servicios de Atención Básica (QualiAB), un instrumento autoadministrado a través de Internet. En total 157 gerentes de Unidades Básicas de Salud de 41 municipios del estado de São Paulo respondieron voluntariamente el QualiAB entre octubre y diciembre de 2014. RESULTADOS: De las 157 unidades, 67 (42,7%) eran unidades salud de la familia y 58 (36,9%) eran unidades básicas de salud de organización "tradicional"; 95 (60,5%) estan ubicadas en una región urbana periférica. En el momento del estudio, ocho (5,0%) unidades no poseían gerente y ocho (5,0%) eran gestionadas por secretarios municipales de salud. Casi el 80% de los gerentes eran enfermeros y desempeñaban múltiples funciones además de la gerencia. En 75 (47,7%) unidades se disponía de apoyo multidisciplinario (supervisión técnica como forma de educación permanente); 60 (38,2%) unidades no contaban con ningún tipo de apoyo multidisciplinario. La participación en procesos de evaluación fue referida por 130 (82,8%) servicios. Las principales modificaciones inducidas por las evaluaciones fueron la planificación y reprogramación de las actividades con participación del equipo multiprofesional en 40 unidades (25,5%) y la definición de un plan anual de actividades en 38 (24,2%). No tuvieron acceso a los resultados de las evaluaciones 29 unidades (17,8%). CONCLUSIÓN: El estudio subraya la importancia de la gestión del trabajo y la necesidad de reinvertir en la formación y valorización de la gestión local como estrategia para hacer efectiva una atención primaria de salud capaz de promover la salud como derecho y condición de ciudadanía.

Importância do gerenciamento local para uma atenção primária à saúde nos moldes de Alma-Ata / Importance of local management for delivery of primary health care according to Alma-Ata principles / Importancia de la gestión local para una atención primaria de salud según las propuestas de Alma-Ata

RESUMO Objetivo Descrever as características da gerência das unidades de atenção primária à saúde e o perfil dos gerentes e discutir as implicações desses elementos para a efetivação dos pressupostos do Sistema Único de Saúde no Brasil de forma coerente com as proposições de Alma-Ata. Métodos Estudo descritivo, transversal, com dados colhidos pelo questionário de Avaliação da Qualidade de Serviços de Atenção Básica (QualiAB), um instrumento autoaplicado via web. O QualiAB foi respondido voluntariamente por 157 gerentes de unidades básicas de saúde de 41 municípios do estado de São Paulo entre outubro e dezembro de 2014. Resultados Das 157 unidades, 67 (42,7%) eram unidades de saúde da família e 58 (36,9%) eram unidades básicas de saúde de organização "tradicional"; 95 (60,5%) se localizavam em região urbana periférica. No momento do estudo, oito (5,0%) unidades não possuíam gerente e oito (5,0%) eram gerenciadas por secretários municipais de saúde. Quase 80% dos gerentes eram enfermeiros e desempenhavam múltiplas funções além da gerência. O matriciamento (supervisão técnica como forma de educação permanente) era feito em 75 (47,7%) unidades; 60 (38,2%) unidades não contavam com nenhum tipo de matriciamento. A participação em processos avaliativos foi referida por 130 (82,8%) serviços. Os principais desdobramentos induzidos por avaliações foram planejamento e reprogramação das atividades com participação da equipe multiprofissional em 40 unidades (25,5%) e definição de um plano anual de atividades em 38 (24,2%). Não tiveram acesso aos resultados das avaliações 29 unidades (17,8%). Conclusão O estudo recoloca a importância da gestão do trabalho e a necessidade de (re)investir na formação e valorização do gerenciamento local como estratégia para efetivar uma atenção primária à saúde capaz de promover a saúde como direito e condição de cidadania.

ABSTRACT Objective To describe the characteristics of the management process in primary health care units as well as the profile of managers, and to discuss the implications of these elements for the attainment of Unified Health System principles in Brazil in accordance with the propositions of the Alma-Ata Declaration. Method This descriptive, cross-sectional study used data collected with the Primary Care Service Quality Assessment tool (QualiAB), a self-administered, web-based instrument. QualiAB was voluntarily answered by 157 unit managers from 41 municipalities in the state of São Paulo from October to December 2014. Results Of 157 units, 67 (42.7%) were family health care units and 58 (36.9%) were "traditional" units; 95 (60.5%) were located in urban peripheries. At the time of the study, eight (5.0%) units did not have a manager and eight (5.0%) were managed by the city health secretary. Almost 80% of the managers were nurses and performed multiple tasks in addition to management. Multidisciplinary support (technical supervision as a means of continuing education) was available in 75 (47.7%) units; 60 (38.2%) units did not have any kind of multidisciplinary support. Participation in evaluative processes was mentioned in 130 (82.8%) units. The main results of evaluations were planning and reprogramming of activities with the engagement of the multiprofessional team in 40 units (25.5%) and definition of an annual activity plan in 38 (24.2%). Twenty-nine units (17.8%) did not have access to the results of evaluations. Conclusion The study supports the importance of work process management and the need to (re)invest in training and recognition of local management as a strategy to produce primary health care that is capable of promoting health as a right and a condition of citizenship.

RESUMEN Objetivo Describir las características de la gestión de las unidades de atención primaria de salud y el perfil de los gerentes, y analizar las implicaciones de esos elementos en la puesta en práctica de los principios del Sistema Único de Salud de Brasil de forma coherente con las propuestas de Alma- Ata. Métodos Estudio descriptivo, transversal, con datos recolectados a través del cuestionario de Evaluación de la Calidad de Servicios de Atención Básica (QualiAB), un instrumento autoadministrado a través de Internet. En total 157 gerentes de Unidades Básicas de Salud de 41 municipios del estado de São Paulo respondieron voluntariamente el QualiAB entre octubre y diciembre de 2014. Resultados De las 157 unidades, 67 (42,7%) eran unidades salud de la familia y 58 (36,9%) eran unidades básicas de salud de organización "tradicional"; 95 (60,5%) estan ubicadas en una región urbana periférica. En el momento del estudio, ocho (5,0%) unidades no poseían gerente y ocho (5,0%) eran gestionadas por secretarios municipales de salud. Casi el 80% de los gerentes eran enfermeros y desempeñaban múltiples funciones además de la gerencia. En 75 (47,7%) unidades se disponía de apoyo multidisciplinario (supervisión técnica como forma de educación permanente); 60 (38,2%) unidades no contaban con ningún tipo de apoyo multidisciplinario. La participación en procesos de evaluación fue referida por 130 (82,8%) servicios. Las principales modificaciones inducidas por las evaluaciones fueron la planificación y reprogramación de las actividades con participación del equipo multiprofesional en 40 unidades (25,5%) y la definición de un plan anual de actividades en 38 (24,2%). No tuvieron acceso a los resultados de las evaluaciones 29 unidades (17,8%). Conclusión El estudio subraya la importancia de la gestión del trabajo y la necesidad de reinvertir en la formación y valorización de la gestión local como estrategia para hacer efectiva una atención primaria de salud capaz de promover la salud como derecho y condición de ciudadanía.

Relato de Experiência da Construção de uma Proposta para Avaliação da Prática Profissional de um Curso Orientado por Competências / Experience Report of Building a Proposal for Evaluation of Professional Practice a Course Oriented Skills

RESUMO O curso de Medicina da Universidade Federal de São Carlos (UFSCar) tem metodologia ativa em seu currículo e proporciona amplas possibilidades de avaliação dos estudantes na prática profissional. No desafio para avaliar a prática profissional de estudantes do curso de Medicina num currículo orientado por competências, criaram-se mecanismos que envolvem o ambiente presencial e à distância, seguindo as Diretrizes Nacionais para os cursos de Medicina e somando o que especialistas internacionais padronizaram como conteúdos essenciais para a avaliação de alunos em sua relação médico-paciente. As competências que os estudantes devem obter vão do âmbito cognitivo ao afetivo e psicomotor. As atividades vão desde a prática médica com preceptores em contextos de diferentes cenários práticos, em diferentes áreas, a atividades em sala de aula – formando um grupo de tutoria com o professor e o Ambiente Virtual de Aprendizagem, realizado na Plataforma Moodle. Todas as atividades da prática profissional da educação médica têm algum grau de dificuldade no momento da avaliação. Apresenta-se uma experiência, realizada em parte no Ambiente Virtual de Aprendizagem, como modelo para estimular o raciocínio clínico-epidemiológico dos estudantes e ajudar os professores no momento da avaliação. Este estudo tem por objetivo mostrar a construção de uma proposta para avaliação da prática profissional do terceiro e quarto anos de um curso de Medicina orientado por competências.

ABSTRACT The medical course at the Federal University of São Carlos (UFSCar) has an innovative curriculum with active methodologies that give students a variety of possibilities for assessment in the practical field. Due to the challenge of evaluating student practice on a medical course with a competence-based curriculum, mechanisms were set up involving the classroom and the internet environment. National guidelines for medical courses were followed, as were standards set by international experts for essential elements in the evaluation of students in terms of their doctor-patient relationship. Student competencies must be assessed in fields of knowledge that range from the cognitive and affective to the psychomotor. The student activities ranged from medical practice with preceptors and contexts of different practical scenarios in different areas, to activities in the classroom, via a tutoring group with the teacher and the Virtual Learning Environment held on the Moodle Platform. All activities used as part of the students’ professional practice involved some degree of difficulty at the time of evaluation. An experiment performed (in part) in the Virtual Learning Environment is outlined as a model to stimulate the students’ clinical and epidemiological reasoning and assist teachers at the time of assessment. This study aims to show the construction of a proposal for the evaluation of professional practice among students in the third and fourth years of medical school guided by competence.

Solvent mimicry with methylene carbene to probe protein topography.

The solvent accessible surface area (SASA) of the polypeptide chain plays a key role in protein folding, conformational change, and interaction. This fundamental biophysical parameter is elusive in experimental measurement. Our approach to this problem relies on the reaction of the minimal photochemical reagent diazirine (DZN) with polypeptides. This reagent (i) exerts solvent mimicry because its size is comparable to water and (ii) shows scant chemical selectivity because it generates extremely reactive methylene carbene. Methylation gives rise to the EM (extent of modification) signal, which is useful for scrutinizing the conformational change triggered by Ca(2+) binding to calmodulin (CaM). The increased EM observed for the full protein is dominated by the enhanced exposure of hydrophobic area in Ca(2+)-CaM. Fragmentation allowed us to quantify the methylene incorporation at specific sites. Peptide 91-106 reveals a major reorganization around the calcium 151 binding site, resulting in local ordering and a greater exposure of the hydrophobic surface. Additionally, this technique shows a high sensitivity to probe recognition between CaM and melittin (Mel). The large decrease in EM indicates the occlusion of a significant hydrophobic area upon complexation. Protection from labeling reveals a larger involvement of the N-terminal and central regions of CaM in this interaction. Despite its smaller size, Mel's differential exposure can also be quantified. Moreover, MS/MS fragmentation realizes the goal of extending the resolution of labeled sites at the amino acid level. Overall, DZN labeling emerges as a useful footprinting method capable of shedding light on physiological conformational changes and interactions.

Alternative cycling modes of the Na(+)/K(+)-ATPase in the presence of either Na(+) or Rb(+).

A comprehensive study of the interaction between Na(+) and K(+) with the Na(+)/K(+)-ATPase requires dissecting the incidence of alternative cycling modes on activity measurements in which one or both of these cations are absent. With this aim, we used membrane fragments containing pig-kidney Na(+)/K(+)-ATPase to perform measurements, at 25°C and pH=7.4, of ATPase activity and steady-state levels of (i) intermediates containing occluded Rb(+) at different [Rb(+)] in media lacking Na(+), and (ii) phosphorylated intermediates at different [Na(+)] in media lacking Rb(+). Most relevant results are: (1) Rb(+) can be occluded through an ATPasic cycling mode that takes place in the absence of Na(+) ions, (2) the kinetic behavior of the phosphoenzyme formed by ATP in the absence of Na(+) is different from the one that is formed with Na(+), and (3) binding of Na(+) to transport sites during catalysis is not at random unless rapid equilibrium holds.

[Assessment of educational actions on the oral health of adolescents within the Family Health Strategy]. / Avaliação das ações de educação na saúde bucal de adolescentes dentro da Estratégia de Saúde da Família.

OBJECTIVE: to evaluate the actions of education on the oral health of adolescents within the scope of the Family Health Strategy. METHODOLOGY: A questionnaire was applied and a clinical exam conducted evaluating biofilm and gingival inflammation in 80 adolescents before and after interventions. A qualitative study of the brushing technique was also performed on 15 adolescents in the sample. RESULTS: there was an improvement in oral hygiene of adolescents who participated in actions at school and an increase in the number of adolescents who had healthy gums or only mild inflammation in the second test in school groups and home visits. With respect to the consumption of candies, both the weekly group and the school group saw a decrease in candy consumption between the first and second tests. There was an increased frequency of brushing in the school group. CONCLUSION: considering the importance of educational activities observed in this study, the process of training and joint planning of activities among the school staff and health professionals would appear to be important.

Avaliação das ações de educação na saúde bucal de adolescentes dentro da Estratégia de Saúde da Família / Assessment of educational actions on the oral health of adolescents within the Family Health Strategy

OBJETIVO: avaliar as ações de educação na saúde bucal de adolescentes dentro da Estratégia de Saúde da Família. METODOLOGIA: foi aplicado um questionário e realizado exame clínico avaliando biofilme e inflamação gengival em 80 adolescentes antes e depois das intervenções. Um estudo qualitativo da técnica de escovação também foi realizado em 15 adolescentes pertencentes à amostra. RESULTADOS: observou-se melhora na higiene oral dos que participaram das ações na escola, aumento no número daqueles que apresentaram gengiva saudável ou inflamação leve no segundo exame nos grupos escola e visita domiciliar. Com relação ao consumo de doces, o grupo semanal e da escola apresentou diminuição entre o 1º e o 2º exames. Houve um aumento na frequência de escovação no grupo escola. CONCLUSÃO: diante da importância das ações educativas verificadas neste estudo, parece-nos importante o processo de capacitação e o planejamento conjunto das ações entre os profissionais da escola e os profissionais de saúde.

OBJECTIVE: to evaluate the actions of education on the oral health of adolescents within the scope of the Family Health Strategy. METHODOLOGY: A questionnaire was applied and a clinical exam conducted evaluating biofilm and gingival inflammation in 80 adolescents before and after interventions. A qualitative study of the brushing technique was also performed on 15 adolescents in the sample. RESULTS: there was an improvement in oral hygiene of adolescents who participated in actions at school and an increase in the number of adolescents who had healthy gums or only mild inflammation in the second test in school groups and home visits. With respect to the consumption of candies, both the weekly group and the school group saw a decrease in candy consumption between the first and second tests. There was an increased frequency of brushing in the school group. CONCLUSION: considering the importance of educational activities observed in this study, the process of training and joint planning of activities among the school staff and health professionals would appear to be important.

E2→E1 transition and Rb(+) release induced by Na(+) in the Na(+)/K(+)-ATPase. Vanadate as a tool to investigate the interaction between Rb(+) and E2.

This work presents a detailed kinetic study that shows the coupling between the E2→E1 transition and Rb(+) deocclusion stimulated by Na(+) in pig-kidney purified Na,K-ATPase. Using rapid mixing techniques, we measured in parallel experiments the decrease in concentration of occluded Rb(+) and the increase in eosin fluorescence (the formation of E1) as a function of time. The E2→E1 transition and Rb(+) deocclusion are described by the sum of two exponential functions with equal amplitudes, whose rate coefficients decreased with increasing [Rb(+)]. The rate coefficient values of the E2→E1 transition were very similar to those of Rb(+)-deocclusion, indicating that both processes are simultaneous. Our results suggest that, when ATP is absent, the mechanism of Na(+)-stimulated Rb(+) deocclusion would require the release of at least one Rb(+) ion through the extracellular access prior to the E2→E1 transition. Using vanadate to stabilize E2, we measured occluded Rb(+) in equilibrium conditions. Results show that, while Mg(2+) decreases the affinity for Rb(+), addition of vanadate offsets this effect, increasing the affinity for Rb(+). In transient experiments, we investigated the exchange of Rb(+) between the E2-vanadate complex and the medium. Results show that, in the absence of ATP, vanadate prevents the E2→E1 transition caused by Na(+) without significantly affecting the rate of Rb(+) deocclusion. On the other hand, we found the first evidence of a very low rate of Rb(+) occlusion in the enzyme-vanadate complex, suggesting that this complex would require a change to an open conformation in order to bind and occlude Rb(+).

Rb(+) occlusion stabilized by vanadate in gastric H(+)/K(+)-ATPase at 25°C.

Despite its similarity with the Na(+)/K(+)-ATPase, it has not been possible so far to isolate a K(+)-occluded state in the H(+)/K(+)-ATPase at room temperature. We report here results on the time course of formation of a state containing occluded Rb(+) (as surrogate for K(+)) in H(+)/K(+)-ATPase from gastric vesicles at 25°C. Alamethicin (a pore-forming peptide) showed to be a suitable agent to open vesicles, allowing a more efficient removal of Rb(+) ions from the intravesicular medium than C(12)E(8) (a non-ionic detergent). In the presence of vanadate and Mg(2+), the time course of [(86)Rb]Rb(+) uptake displayed a fast phase due to Rb(+) occlusion. The specific inhibitor of the H(+)/K(+)-ATPase SCH28080 significantly reduces the amount of Rb(+) occluded in the vanadate-H(+)/K(+)-ATPase complex. Occluded Rb(+) varies with [Rb(+)] according to a hyperbolic function with K(0.5)=0.29±0.06mM. The complex between the Rb(+)-occluded state and vanadate proved to be very stable even after removal of free Mg(2+) with EDTA. Our results yield a stoichiometry lower than one occluded Rb(+) per phosphorylation site, which might be explained assuming that, unlike for the Na(+)/K(+)-ATPase, Mg(2+)-vanadate is unable to recruit all the Rb(+)-bound to the Rb(+)-occluded form of the Rb(+)-vanadate-H(+)/K(+)-ATPase complex.

Co-infecção por Nocardia asteroides e complexo Mycobacterium avium em paciente infectado pelo HIV – Relato de caso / Nocardia asteroides and Mycobacterium avium complex co-infection in a HIV patient: Case report

Desde o início da descoberta do vírus da imunodeficiência humana (HIV), as infecções pulmonares têm sido a principal causa de morbidade e mortalidade entre os pacientes infectados pelo vírus. Nocardia sp é considerado um patógeno oportunista entre estes pacientes; entretanto, quando a infecção acontece, o pulmão é o órgão mais freqüentemente acometido, podendo ocorrer lesões cavitárias. Infecção pulmonar pelo complexo Mycobacterium avium não é comum e raramente está associada com cavitação. Casos de co-infecção por Nocardia sp e complexo Mycobacterium avium em pacientes portadores do HIV são raros. Relatamos um caso de infecção pulmonar em paciente com sorologia positiva para HIV. A cultura de escarro revelou a presença de Nocardia asteroides e complexo Mycobacterium avium.

Características da doença periodontal em hanseníase / Characteristics of periodontal disease in leprosy

A hanseníase é uma doença inflamatória, crônica, granulomatosa, causada pelo Mycobacterium leprae. Ainda endêmica em alguns países em desenvolvimento, seus principais sinais clínicos são lesões de pele hipoestésicas e espessamento dos nervos periféricos. As áreas da cavidade bucal mais comumente afetadas são o palato duro, o palato mole, a úvula, os lábios, a língua e a gengiva. Entende-se por doença periodontal um conjunto de condições inflamatórias, de caráter crônico e de origem bacteriana, que começa afetando o tecido gengival e pode levar à perda dos tecidos de suporte do elemento dental. Com o objetivo de estabelecer a prevalência da doença periodontal em pacientes portadores das diferentes formas de hanseníase, foram examinados 100 pacientes hansenianos do Instituto Lauro de Souza Lima, em Bauru (SP), submetidos a exames de sondagem periodontal, nível de inserção clínica, índice de placa bacteriana, presença de sangramento gengival à sondagem, mobilidade dentária e exame radiográfico periapical como método complementar de diagnóstico. As análises estatísticas demonstraram que, assim como na população hígida, a doença periodontal foi marcadamente prevalente em adultos e idosos e em fumantes e não se encontrou nenhuma evidência de que a hanseníase seja determinante para ocorrência de doença periodontal.

Perfil epidemiológico, clínico e evolutivo da tuberculose na região de Bauru-SP / Epidemiological, clinical and evolutionary profile of tuberculosis in the Bauru-SP region

Trata o presente estudo da descriçao das características epidemiológicas, clínicas e evolutivas da tuberculose na Regiao de Governo de Bauru-SP. Foi realizado com informaçoes extraídas das fichas de notificaçao da doença e dos registros acerca da situaçao no sétimo mês após o início do tratamento dos 865 casos, que constituíram o universo dos doentes notificados de julho de 1992 a dezembro de 1995. Verificou-se decréscimo, tanto do número de notificaçoes, quanto dos coeficientes de incidência, no decorrer dos anos estudados. A incidência da tuberculose declinou de 53,6 para 45,1 casos por 100.000 habitantes, de 1993 para 1995. Nao obstante, no mesmo período, aumentou a ocorrência em infectados pelo HIV e em presidiários