Development and validation of a simple and rapid capillary zone electrophoresis method for determination of nnrti nevirapine in pharmaceutical formulations

A simple and fast capillary zone electrophoresis (CZE) method has been developed and validated for quantification of a non-nucleoside reverse transcriptase inhibitor (NNRTI) nevirapine, in pharmaceuticals. The analysis was optimized using 10 mmol L-1 sodium phosphate buffer pH 2.5, +25 kV applied voltage, hydrodynamic injection 0.5 psi for 5 s and direct UV detection at 200 µm. Diazepam (50.0 µg mL-1) was used as internal standard. Under these conditions, nevirapine was analyzed in approximately less than 2.5 min. The analytical curve presented a coefficient of correlation of 0.9994. Limits of detection and quantification were 1.4 µg mL-1 and 4.3 µg mL-1, respectively. Intra- and inter-day precision expressed as relative standard deviations were 1.4% and 1.3%, respectively and the mean recovery was 100.81%. The active pharmaceutical ingredient was subjected to hydrolysis (acid, basic and neutral) and oxidative stress conditions. No interference of degradation products and tablet excipients were observed. This method showed to be rapid, simple, precise, accurate and economical for determination of nevirapine in pharmaceuticals and it is suitable for routine quality control analysis since CE offers benefits in terms of quicker method development and significantly reduced operating costs.

Ecologia de Anopheles (Nyssorhynchus) darlingi Root em área de implantação de empreendimento hidrelétrico, na divisa dos Estados do Mato Grosso do Sul e São Paulo

INTRODUÇÃO: Hidrelétricas alteram o fluxo das águas e provocam impactos sobre a composição de mosquitos, justificando-se essa pesquisa. O objetivo da pesquisa foi estudar anofelinos de área sob a influência de um novo lago e avaliar a vulnerabilidade relativa à malária. MÉTODOS: Foram feitas coletas de Anopheles nas margens da Represa Porto Primavera, durante as fases do alagamento até sua cota máxima. Utilizaram-se as técnicas: atrativa humana, de armadilha de Shannon e concha entomológica. Os indicadores Riqueza e Diversidade foram utilizados para medir o impacto. A análise das distribuições temporais foi realizada pelo teste Mann-Whitney, considerando localidade, cota e método de captura como variáveis independentes (α=0,05). RESULTADOS: A densidade de Anopheles darlingi oscilou entre as localidades A, B e C, sendo que os maiores picos foram para B e C. Com a estabilidade do lago, no último nível, evidenciou-se a tendência de redução da densidade de Anopheles darlingi. CONCLUSÕES: Sugere-se que o risco de autoctonia de malária nas proximidades do lago permanece inalterado, ficando o alerta para esporádicas infecções humanas.

Validation of a food frequency questionnaire to assess the consumption of carotenoids, fruits and vegetables among adolescents: the method of triads

The aim of this study was to validate the intake of carotenoids, fruits and vegetables estimated by the Food Frequency Questionnaire for Adolescents (FFQA) using the method of triads. Blood samples were collected from 80 elementary school adolescents to assess serum levels of β-carotene. Partial correlation coefficients (r) were calculated between an estimated intake of carotenoids, fruits and vegetables and the serum levels of β-carotene. Validity coefficients were calculated using the method of triads. With the exception of carotenoids, partial r from the food frequency questionnaire (FFQ) were greater than those of the 24-hour recall (24hR). The fruit/vegetable group showed the highest partial r for the FFQ (r = 0.235) and the 24hR (r = 0.137). The highest validity coefficient was obtained for the vegetable group, as assessed by the FFQ (r = 0.873). On average, the validity coefficient values for the FFQ were greater than those obtained for the 24hR or the β-carotene serum levels. The FFQA is an accurate tool for estimating the intake of carotenoids, fruits and vegetables in this population group.

Influência do consumo alimentar na gestação sobre a retenção de peso pós-parto

OBJETIVO: Avaliar a influência da alimentação durante a gestação sobre a retenção de peso pós-parto. MÉTODOS: Foram acompanhadas 82 gestantes adultas e saudáveis que iniciaram o pré-natal em serviço público de saúde no Município de São Paulo, SP, entre abril e junho de 2005. As medidas de peso e estatura foram aferidas na primeira entrevista (até 16 semanas de gestação) e a medida de peso foi repetida em visita domiciliar 15 dias após o parto. O Recordatório de 24 horas foi usado para avaliar o consumo alimentar e foi aplicado nos três trimestres da gestação. Foi calculado o consumo médio de gordura saturada, fibras, açúcar adicionado, refrigerantes, alimentos processados, frutas, verduras e legumes, e a densidade energética da dieta. A retenção de peso foi obtida pela diferença entre a medida de peso pós-parto e a primeira medida realizada. A influência da alimentação sobre a retenção de peso pós-parto foi avaliada por meio de análise de regressão linear múltipla e foi realizado o teste para tendência linear. As variáveis utilizadas para ajuste do modelo foram: índice de massa corporal no início da gestação, estatura, renda familiar per capita, tabagismo, idade e escolaridade. RESULTADOS: O índice de massa corporal inicial médio foi de 24 kg/m² e a retenção média de peso foi de 1,9 kg. O aumento do consumo de gordura saturada (p = 0,005) e alimentos processados (p = 0,014) elevou de forma significativa a retenção de peso pós-parto, após ajuste pelas variáveis de controle. As demais variáveis de consumo alimentar não apresentaram relação significativa com a variável desfecho. CONCLUSÕES: O maior consumo de alimentos não saudáveis, como alimentos processados, e de gordura saturada influencia a elevação da retenção de peso pós-parto.

Validação do Questionário de Freqüência Alimentar para Adolescentes (QFAA) por grupos de alimentos em uma população de escolares

OBJETIVO: Verificar a validade do Questionário de Freqüência Alimentar para Adolescentes para avaliar o consumo de grupos de alimentos entre escolares de Piracicaba, São Paulo. MÉTODOS: Participaram do estudo 94 adolescentes, com idade entre 11 e 15 anos, matriculados em uma escola da rede pública. O consumo alimentar foi avaliado pelo Questionário de Freqüência Alimentar para Adolescentes (QFAA) e a média de dois Recordatórios de 24 horas (R24h) foi utilizada como método de referência. Os itens alimentares foram classificados em 18 grupos. Foram realizadas análises descritivas, teste t-Student pareado e de Wilcoxon, coeficientes de correlação de Pearson e de Spearman. Foram também utilizadas análise de quartis e estatística Kappa ponderado. Os coeficientes de correlação foram corrigidos pela variância intrapessoal dos R24h, estimada a partir de ANOVA com um fator de classificação. RESULTADOS: Não foram verificadas diferenças significativas entre os instrumentos para o arroz, massas, carnes, refrigerantes e sucos artificiais. Os coeficientes de correlação corrigidos pela variabilidade intrapessoal variaram de -0,26 a 0,78. A concordância de classificação dos indivíduos no mesmo quartil de consumo para ambos os métodos variou de 22% (massas) a 50% (feijão). Para quartis opostos, os grupos que tiveram mais de 10% dos indivíduos classificados incorretamente foram massas (19%), carnes (13%) e gorduras (11%). Os valores de Kappa ponderado variaram de - 0,15 (massas) a 0,56 (feijão). O QFAA superestimou o consumo de quase a totalidade dos grupos alimentares e subestimou os grupos dos óleos, feijão, carnes e refrigerantes. CONCLUSÃO: O instrumento apresentou boa validade para feijão, verduras e legumes, leite e derivados, biscoitos recheados e para o arroz.

O uso do tempo no cotidiano de mulheres idosas: um método indicador do estilo de vida de grupos populacionais

O estudo do uso do tempo considera a heterogeneidade do envelhecimento, analisando-o multifatorialmente, permitindo vislumbrar o estilo de vida de indivíduos idosos. Este estudo objetivou descrever o uso do tempo de 75 idosas (68,04 ± 8,36 anos), através das suas atividades diárias. Instrumentos utilizados: teste de cognição (Clock Completion Test), formulário para dados pessoais e entrevista estruturada (Time Diary) para relato das atividades diárias. Para decodificação e classificação das atividades diárias, utilizou-se a classificação australiana para estudos de uso do tempo, distribuindo-as em nove grupos de atividades principais. Foram verificados os contextos físico (local) e social (parceiros sociais) das atividades. Verificou-se que grande parte do tempo destinou-se às atividades obrigatórias (atividades domésticas e de cuidados pessoais). A maior proporção do tempo livre destinou-se ao lazer passivo (assistir televisão) com pouco envolvimento em atividades físicas. A casa e estar com membros da família ou sozinhas representaram o contexto físico e social mais presentes. O estudo permitiu um vislumbre do estilo de vida do grupo. Provavelmente houve influência de fatores individuais como idade, gênero, grau de instrução, estado civil e nível socioeconômico sobre os padrões encontrados para o uso do tempo.

Evaluation of oviposition traps as an entomological surveillance method for Aedes aegypti (Diptera, Culicidae)

This study aimed to describe the behavior of oviposition traps for Aedes aegypti over time, to compare it with the larval survey and to investigate the association with climatic variables. It was conducted in São José do Rio Preto city, São Paulo. Daily climatic data and fortnightly measurements for oviposition traps and larval infestation were collected from October 2003 to September 2004. Three different periods were identified in the behavior of oviposition traps' positivity and mean number of eggs: increase, plateau and decrease in values. These measurements followed the variation of climatic data from the first and third periods. High correlation was obtained between the positivity and the mean number of eggs. The oviposition traps showed higher capacity to detect the vector than did larval survey. It was observed that the first (October to December) and third (May to September) periods were considered to be the most suitable to use oviposition traps than larval surveys.

Pathogenicity of different rabies virus isolates and protection test in vaccinated mice

This study was aimed to evaluate and compare the pathogenicity of rabies virus isolated from bats and dogs, and to verify the efficacy of a commercial rabies vaccine against these isolates. For evaluation of pathogenicity, mice were inoculated by the intramuscular route (IM) with 500MICLD50/0.03mL of the viruses. The cross-protection test was performed by vaccinating groups of mice by the subcutaneous route and challenged through the intracerebral (IC) route. Isolates were fully pathogenic when inoculated by the IC route. When inoculated intramuscularly, the pathogenicity observed showed different death rates: 60.0% for the Desmodus rotundus isolate; 50.0% for dog and Nyctinomops laticaudatus isolates; 40.0% for Artibeus lituratus isolate; 9.5% Molossus molossus isolate; and 5.2% for the Eptesicus furinalis isolate. Mice receiving two doses of the vaccine and challenged by the IC route with the isolates were fully protected. Mice receiving only one dose of vaccine were partially protected against the dog isolate. The isolates from bats were pathogenic by the IC route in mice. However, when inoculated through the intramuscular route, the same isolates were found with different degrees of pathogenicity. The results of this work suggest that a commercial vaccine protects mice from infection with bat rabies virus isolates, in addition to a canine rabies virus isolate.

Rapid detection of bovine coronavirus by a semi-nested RT-PCR

Bovine coronavirus (BCoV) is a member of the group 2 of the Coronavirus (Nidovirales: Coronaviridae) and the causative agent of enteritis in both calves and adult bovine, as well as respiratory disease in calves. The present study aimed to develop a semi-nested RT-PCR for the detection of BCoV based on representative up-to-date sequences of the nucleocapsid gene, a conserved region of coronavirus genome. Three primers were designed, the first round with a 463bp and the second (semi-nested) with a 306bp predicted fragment. The analytical sensitivity was determined by 10-fold serial dilutions of the BCoV Kakegawa strain (HA titre: 256) in DEPC treated ultra-pure water, in fetal bovine serum (FBS) and in a BCoV-free fecal suspension, when positive results were found up to the 10-2, 10-3 and 10-7 dilutions, respectively, which suggests that the total amount of RNA in the sample influence the precipitation of pellets by the method of extraction used. When fecal samples was used, a large quantity of total RNA serves as carrier of BCoV RNA, demonstrating a high analytical sensitivity and lack of possible substances inhibiting the PCR. The final semi-nested RT-PCR protocol was applied to 25 fecal samples from adult cows, previously tested by a nested RT-PCR RdRp used as a reference test, resulting in 20 and 17 positives for the first and second tests, respectively, and a substantial agreement was found by kappa statistics (0.694). The high sensitivity and specificity of the new proposed method and the fact that primers were designed based on current BCoV sequences give basis to a more accurate diagnosis of BCoV-caused diseases, as well as to further insights on protocols for the detection of other Coronavirus representatives of both Animal and Public Health importance.

Anticorpos anti Brucella abortus, anti Brucella canis e anti Leptospira spp. em raposas (Pseudalopex vetulus) do semiárido paraibano, Nordeste do Brasil

O objetivo do presente trabalho foi determinar a ocorrência de anticorpos anti Brucella abortus, anti Brucella canis e anti Leptospira spp. em raposas (Pseudalopex vetulus). Para tanto, foram utilizadas 60 raposas atropeladas em rodovias no semiárido da Paraíba, Nordeste do Brasil. Para a detecção de anticorpos anti Brucella abortus, o teste do antígeno acidificado tamponado (AAT) foi empregado como teste de triagem, e a prova do 2-mercaptoetanol foi empregada como método confirmatório. Para o diagnóstico sorológico das infecções por Brucella canis e Leptospira spp., foram utilizados os testes de imunodifusão em gel de agar (IDGA) e soroaglutinação microscópica, respectivamente. Todas as amostras foram negativas na pesquisa de anticorpos anti Brucella canis e anti Leptospira spp. Das 60 raposas testadas, 16 (26,6%) foram positivas para anticorpos anti Brucella abortus no teste de AAT, e quatro (6,7%) amostras foram confirmadas no teste de 2-mercaptoetanol, sendo duas amostras com título 100 e duas com título 50.

The ability of the BANA test to detect different levels of P. gingivalis, T. denticola and T. forsythia

The aim of this study was to evaluate the ability of the BANA Test to detect different levels of Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia or their combinations in subgingival samples at the initial diagnosis and after periodontal therapy. Periodontal sites with probing depths between 5-7 mm and clinical attachment level between 5-10 mm, from 53 subjects with chronic periodontitis, were sampled in four periods: initial diagnosis (T0), immediately (T1), 45 (T2) and 60 days (T3) after scaling and root planing. BANA Test and Checkerboard DNA-DNA hybridization identified red complex species in the subgingival biofilm. In all experimental periods, the highest frequencies of score 2 (Checkerboard DNA-DNA hybridization) for P. gingivalis, T. denticola and T. forsythia were observed when strong enzymatic activity (BANA) was present (p < 0.01). The best agreement was observed at initial diagnosis. The BANA Test sensitivity was 95.54% (T0), 65.18% (T1), 65.22% (T2) and 50.26% (T3). The specificity values were 12.24% (T0), 57.38% (T1), 46.27% (T2) and 53.48% (T3). The BANA Test is more effective for the detection of red complex pathogens when the bacterial levels are high, i.e. in the initial diagnosis of chronic periodontitis.

An Environmental Friendly Procedure for Photometric Determination of Hypochlorite in Tap Water Employing a Miniaturized Multicommuted Flow Analysis Setup

A photometric procedure for the determination of ClO(-) in tap water employing a miniaturized multicommuted flow analysis setup and an LED-based photometer is described. The analytical procedure was implemented using leucocrystal violet (LCV; 4,4', 4 ''-methylidynetris (N, N-dimethylaniline), C(25)H(31)N(3)) as a chromogenic reagent. Solenoid micropumps employed for solutions propelling were assembled together with the photometer in order to compose a compact unit of small dimensions. After control variables optimization, the system was applied for the determination of ClO(-) in samples of tap water, and aiming accuracy assessment samples were also analyzed using an independent method. Applying the paired t-test between results obtained using both methods, no significant difference at the 95% confidence level was observed. Other useful features include low reagent consumption, 2.4 mu g of LCV per determination, a linear response ranging from 0.02 up to 2.0 mg L(-1) ClO(-), a relative standard deviation of 1.0% (n = 11) for samples containing 0.2 mg L(-1) ClO(-), a detection limit of 6.0 mu g L(-1) ClO(-), a sampling throughput of 84 determinations per hour, and a waste generation of 432 mu L per determination.

Energy system contributions to the special judo fitness test

Purpose: This study investigated the energy system contributions of judo athletes to the Special Judo Fitness Test (SJFT). Methods: Fourteen male judo athletes performed the SJFT, which comprised three periods of judo activity (A = 15 s, B and C = 30 s) interspersed with 10 s rest intervals. During this test, one athlete threw two others positioned 6 m from each other using the ippon-seoi-nage technique. The fractions of the aerobic, anaerobic alactic and anaerobic lactic systems were calculated based on oxygen uptake, the fast component of excess postexercise oxygen uptake, and changes in net blood lactate, respectively. The contribution of the three energy systems was compared using a repeated measures analysis of variance and Bonferroni's multiple comparisons test. Compound symmetry, or sphericity, was determined by Mauchly's test. A level of significance of 5% (P < .05) was adopted in all analyses. Results: The alactic energy system presented a higher (F = 20.9; P < .001; power observed = 1.0) contribution (86.8 +/- 23.6 kJ; 42.3 +/- 5.9%) during the test when compared with both aerobic (57.1 +/- 11.3 kJ; 28.2 +/- 2.9%) and lactic (58.9 +/- 12.1 kJ; 29.5 +/- 6.2%) energy systems (P < .001 for both comparisons). Conclusions: The higher alactic contribution seems to be a consequence of the high-intensity efforts performed during the test, and its intermittent nature. Thus, when using the SJFT, coaches are evaluating mainly their athletes' anaerobic alactic system, which can be considered to be the most predominant system contributing to the actions (techniques) performed in the match.

The role of aerobic fitness on session rating of perceived exertion in futsal players

Purpose: The aim of this study was to verify the influence of aerobic fitness (VO(2)max) on internal training loads, as measured by the session rating of perceived exertion (session-RPE) method. Methods: Nine male professional outfield futsal players were monitored for 4 wk of the in-season period with regards to the weekly accumulated session-RPE, while participating in the same training sessions. Single-session-RPE was obtained from the product of a 10-point RPE scale and the duration of exercise. Maximal oxygen consumption was determined during an incremental treadmill test. Results: The average training load throughout the 4 wk period varied between 2,876 and 5,035 arbitrary units. Technical-tactical sessions were the predominant source of loading. There was a significant correlation between VO(2)max (59.6 +/- 2.5 mL.kg(-1).min(-1)) and overall training load accumulated over the total period (r = -0.75). Conclusions: The VO(2)max plays a key role in determining the magnitude of an individual's perceived exertion during futsal training sessions.