Análise metagenômica da microbiota de ambientes aquáticos do estado do Rio Grande do Norte - Brasil

The screening for genes in metagenomic libraries from soil creates opportunities to explore the enormous genetic and metabolic diversity of microorganisms. Rivers are ecosystems with high biological diversity, but few were examined using the metagenomic approach. With this objective, a metagenomic library was constructed from DNA soil samples collected at three different points along the Jundiaí-river (Rio Grande do Norte-Brazil). The points sampled are from open area, rough terrain and with the direct incidence of sunlight. This library was analyzed functionally and based in sequence. For functional analysis Luria-Bertani solid medium (LB) with NaCl concentration varied from 0.17M to 0.85M was used for functional analysis. Positives clones resistant to hypersaline medium were obtained. The recombinant DNAs were extracted and transformed into Escherichia coli strain DH10B and survival curves were obtained for quantification of abiotic stress resistance. The sequences of clones were obtained and submitted to the BLASTX tool. Some clones were found to hypothetical proteins of microorganisms from both Archaea and Bacteria division. One of the clones showed a complete ORF with high similarity to glucose-6-phosphate isomerase which participates in the synthesis of glycerol pathway and serves as a compatible solute to balance the osmotic pressure inside and outside of cells. Subsequently, in order to identify genes encoding osmolytes or enzymes related halotolerance, environmental DNA samples from the river soil, from the water column of the estuary and ocean were collected and pyrosequenced. Sequences of osmolytes and enzymes of different microorganisms were obtained from the UniProt and used as RefSeqs for homology identification (TBLASTN) in metagenomic databases. The sequences were submitted to HMMER for the functional domains identification. Some enzymes were identified: alpha-trehalose-phosphate synthase, L-ectoina synthase (EctC), transaminase L-2 ,4-diaminobutyric acid (EctB), L-2 ,4-diaminobutyric acetyltransferase (EctA), L-threonine 3 dehydrogenase (sorbitol pathway), glycerol-3-phosphate dehydrogenase, inositol 3-phosphate dehydrogenase, chaperones, L-proline, glycine betaine binding ABC transporter, myo-inositol-1-phosphate synthase protein of proline simportadora / PutP sodium-and trehalose-6-phosphate phosphatase These proteins are commonly related to saline environments, however the identification of them in river environment is justified by the high salt concentration in the soil during prolonged dry seasons this river. Regarding the richness of the microbiota the river substrate has an abundance of halobacteria similar to the sea and more than the estuary. These data confirm the existence of a specialized response against salt stress by microorganisms in the environment of the Jundiaí river

Metagenômica: busca de novos genes envolvidos com a biodegradação de hidrocarbonetos

Industrial activities, oil spills and its derivatives, as well as the incomplete combustion of fossil fuels have caused a great accumulation of hydrocarbons in the environment. The number of microorganisms on the planet is estimated at 1030 and prokaryotes the most abundant. They colonized diverse environments for thousands of years, including those considered extreme and represent an untapped source of metabolic and genetic diversity with a large biotechnological potential. It is also known that certain microorganisms have the enzymatic capacity to degrade petroleum hydrocarbons and, in many ecosystems, there is an indigenous community capable of performing this function. The metagenomic has revolutionized the microbiology allowing access uncultured microbial communities, being a powerful tool for elucidation of their ecological functions and metabolic profiles, as well as for identification of new biomolecules. Thus, this study applied metagenomic approaches not only for functional selection of genes involved in biodegradation and emulsification processes of the petroleum-derived hydrocarbons, but also to describe the taxonomic and metabolic composition of two metagenomes from aquatic microbiome. We analyzed 123.116 (365 ± 118 bp) and 127.563 sequences (352 ± 120 bp) of marine and estuarine metagenomes, respectively. Eight clones were found, four involved in the petroleum biodegradation and four were able to emulsify kerosene indicating their abilities in biosurfactants synthesis. Therefore, the metagenomic analyses performed were efficient not only in the search of bioproducts of biotechnological interest and in the analysis of the functional and taxonomic profile of the metagenomes studied as well

Proteínas SAG1, SAG2, SAG3 de toxoplasma gondii como perspectivas para o desenvolvimento de protótipo vacinal contra a toxoplasmose

Toxoplasmosis is a zoonosis of worldwide distribution caused by the protozoan Toxoplasma gondii, triggering dangerous complications in immunocompromised patients and pregnant women, as well as having great economic impact for the livestock. So far the control of toxoplasmosis is made primarily by chemotherapy. However, most drugs used routinely have some limitations. In order to control this disease, several research groups, including ours, has been working to develop a medical-veterinary vaccine based on parasite antigens, vectors and protocols of immunization. In this study were implemented and standardized methodologies for amplification and cloning of recombinant immunogens in the system for the development of a prototype vaccine, based on the surface antigens of T. gondii and recombinant adenovirus encoding these antigens. Genes encoding BAG1, GRA2 and SAG1 proteins were amplified. We established a strategy for cloning SAG1, SAG2, SAG3 and TgAMA1- genes in recombinant system. The genes encoding SAG1 and SAG2 were cloned and their sequences showed high similarity with sequences from GenBank. The virtual translation of these proteins showed polymorphisms in the amino acid sequence, which can be correlated with levels of antigenicity. Simultaneously, the adenovirus encoding the SAGs (HAdSAGs) were expanded, purificated and characterizated. Immunization of C57bl/6 mice, using viral supernatant was not enought to elicit immune responses at high levels, being required HAdSAGs titration for future immunizations. Therefore, this study allowed the cloning of the two genes important for the development of a prototype vaccine. Besides, implementations methodologies that permit advancements in the development of a vaccine against toxoplasmosis using adenovirus to express proteins of the parasite

Acesso aos serviços odontológicos e incidência de cárie em adolescentes e fatores de risco em município do Nordeste Brasileiro, 2006

The aims of this study were to analyze the access of dental services by child population, to determine the prevalence of dental caries, gingivitis and malocclusion in resident children from the municipal district of Sobral Ceará and to evaluate the incidence of the dental decay in adolescents associated with the factors related to socioeconomic condition, access to health services and self-perception. This study had as main factor the multidisciplinary represented by the participation of health professional (doctors, dentists, nurses) in the development of the survey's initial reference; student from Human Sciences area to apply the structured questionnaire in domiciliary visits; statistics professionals in the orientation of the analysis to be held and family health team (community health agents, dentists and dental clinic assistants) in the scheduling of domiciliary visits and the accomplishment of oral exam. The sample was determined from the domicile record that included children born between 1990 and 1994 to develop the research Children health conditions in the municipal district of Sobral Ceará . The first sample comprised 3425 parents of children from 5 to 9 years old, living in the urban area at the municipal district of Sobral Ceará, aiming at identifying the most important factors associated to the access to dental service. From this sample, 1021 children were selected in a systematic way, for the accomplishment to the epidemiological study of decay, gingivitis and malocclusion. In the study's third phase, in order to arrange the group to be followed, 688 adolescents were examined and interviewed, by means of the active search from the 1021 individuals that had been previously examined. It was observed that 50.9% of the children had access to dental service at least once in a lifetime. Of this total, 65.3% accomplished it during the last year, and 85.4% of these did in public services, what allows to identify the importance of this sector in the access to dental services. It was observed that the factors that most affected the access to dental 129 services were related to socioeconomic condition, such as the access to health plan, the possession of toothbrush, garbage collecting, mother s schooling, sewerage treatment and malnutrition. In relation to oral diseases, an increase in the DMF-T index according the age was observed, from 0.10 in five years old to 1.66 in the nine years old, while with the dmf-t index, the inverse happened, since the index decreased from 3.59 in five years old to 2.69 in nine years old. In relation to gingivitis, an average 32.7% of the children presented gum bleeding. In what concerns malocclusion, it was observed that 60.3% of the children didn't present any problem, 30.17% had light malocclusion and 9.5% severe malocclusion. The average incidence of dental caries was 1.86 teeth per youngster. Among the studied variables, tooth pain in the last six months, mother's income and school snack, adjusted by the perception about the need of treatment, the mother's schooling and the dentist's appointment at least once in a lifetime, were the variables that presented positive relationship with the high incidence of dental caries on this population by logistic regression. Variables of socioeconomic nature, related to the access to health services and behavior and biological variables presented a relationship with the high caries incidence. The study point out to the need of developing health actions in a humanized way, by an oral health team effectively bound to the population's interest, with the great objective to provide, with the public health services managers, adequate conditions to improve oral health

Apitidão funcional e comportamentos relacionados à saúde em mulheres idosas do município de Jequié-BA

The identification of the factors that interfere in the decline of functional conditions is useful in the planning of actions addressing the improvement in the conditions of the lives of elderly people. The purpose of this investigation was to analyze the relationship between social demographics and health aspects of the functional condition in elderly women of low income of the Brazilian northeast. This crosssectional study involved a representative sample of 222 women with an average age of 70 years (± 7.1), belonging to coexisting groups and that were resident in the urban area of the municipal district of Jequié /Bahia. In order to achieve this objective, a battery of physical tests of functional aptitude was carried out previously tested in pilot study, anthropometric measurements collected with a comparison of the measures referred to the reported weight and height as well as the application of an interview with questions containing subjects related to social demographic variables, clinical conditions and health, physical conditions and behaviors. Descriptive statistics Proceedings (frequency, average, standard deviation and percent distribution) were used for statistic analysis, and the calculation of the respective odds ratio by binary logistics regression, for the analysis of factors hierarchically grouped; p<0.05. The prevalence of 56% (n=122) of women considered with moderated or serious type of functional limitations was found, In which from multi-varied hierarchical analysis, significant association was verified with the age group over 80 years (p=0.02), conditions of widowhood (p=0.04), presence of arterial hypertension (p=0.001), and physical inactivity during leisure time (p=0.03). On the other hand for functional incapacities the prevalence was of 46.8% (n=104) being associated to the increase of the age (p=0.01), hospitalization (p=0.02), absence of physical activities along their lives (p=0.001) and the occurrence of alterations in the cognitive function (p=0.001). The normative table for the parameters of physical fitness generated conducive to health professionals in the diagnosis of health conditions and the prescription of physical exercises. The identified characteristics that are associated with the functional limitations / functional incapacities suggest a complex causal net in the determination of the functional condition in elderly women. However, actions addressed to the incentive of the practice of physical activities in the leisure time and the preservation of the cognitive function can contribute to a life with more quality for these people. This research was multidisciplinary approach to involve elements of psychology, nutrition and Physical Education in the elucidation of the object of study related to the functional condition of elderly women

Fatores preditivos da dor em pacientes submetidos à cirurgia cardíaca

A identificação de fatores que interferem na dor pós-operatória é útil para minimizar o sofrimento desnecessário e favorecer a uma intervenção analgésica adequada, evitando generalizações nas condutas terapêuticas. O propósito dessa investigação foi identificar os fatores preditivos da dor em pacientes submetidos à cirurgia cardíaca e a relação existente entre dor, analgesia e personalidade. Trata-se de um estudo prospectivo e analítico, aprovado pelo comitê de ética da UFRN (175/06), o qual proporcionou uma abordagem multidisciplinar ao envolver áreas distintas como: fisioterapia, psicologia, médica e enfermagem (interdisciplinaridade) na elucidação do objeto de estudo relacionado a fatores preditivos da dor. Para caracterização geral dos pacientes foi utilizada uma ficha de avaliação fisioterapêutica; a dor pós-operatória foi avaliada pela escala numérica de dor e questionário para dor McGill e o Inventário Millon de Estilos de Personalidade (MIPS) foi utilizado para identificar e avaliar as manifestações das características comportamentais e de personalidade. Foram acompanhados, do 1º ao 5º dia de pós-operatório (DPO), 160 pacientes submetidos à cirurgia cardíaca, 57,5% do sexo masculino, com idade média de 56,8±14,4 anos, sendo incluídos no estudo aqueles que se queixaram de dor no pós-operatório em pelo menos um dos dias de avaliação e assinaram termo de concentimento livre e esclarecido (TCLE). Os dados foram submetidos ao teste Qui-quadrado, regressão logística multivariada, teste de correlação de Spearman, teste t e ANOVA. Ao serem submetidos à análise de regressão foram encontrados seis fatores preditivos da dor: tempo cirúrgico > 3 horas, dreno mediastinal e lateral, tosse, vômitos, tempo de dreno > 24 horas e sexo feminino. Estes fatores apresentaram uma correlação positiva e significava com a intensidade dolorosa referida pelos pacientes na escala numérica de dor e os pacientes que apresentaram mais fatores preditivos da dor referiram mais dor. 23 Já em relação ao McGill não se obteve diferença significativa entre os pacientes com mais e menos fatores preditivos. Observou-se que foram administrados diferentes tipos de analgésicos, isolados ou associados, sendo estes: paracetamol, paracetamol associado à codeína, dipirona, tramadol, toradol e tilatil. A percepção dolorosa apresentou magnitudes variando de leve a moderada do 1º ao 5º DPO e ao ser relacionada com as características de personalidade, nos pacientes com menos dor, observou-se os fatores: preservação, individualismo, introversão e os com mais dor foram: proteção, extroversão, retraimento, discrepância, afetividade, acomodação, comunicabilidade e firmeza. Evidenciou-se que o comportamento doloroso pós-cirurgia cardíaca é multifatorial e que a determinação da existência de fatores preditivos da dor permite ao profissional da área de saúde fazer o uso adequado dos analgésicos, haja vista que o alívio da dor é responsabilidade de todos os profissionais da saúde. Os aspectos psicológicos enquanto características de personalidade podem influenciar padrões de comportamento como os observados.

Atividade física e outros fatores associados à função sexual em mulheres de meia idade

Este estudo teve por objetivo avaliar os fatores associados à disfunção sexual em mulheres de meia idade. Realizou-se um estudo descritivo transversal, que compreendeu 370 mulheres, entre 40 a 65 anos, atendidas nas Unidades Básicas de Saúde de cada distrito sanitário (Norte, Sul, Leste e Oeste) da cidade de Natal, no estado do Rio Grande do Norte, Brasil. Para avaliar a função sexual utilizou-se o Female Sexual Function Index (FSFI). A sintomatologia climatérica foi avaliada por meio do Menopause Rating Scale (MRS). O Índice de Blatt-Kupperman (IMBK) foi utilizado para avaliação quantitativa global da ocorrência de sintomas/queixas. A atividade física foi avaliada pelo questionário International Physical Activity Questionnaire - IPAQ (versão curta). A avaliação da qualidade de vida geral se deu pelo WHOQOL-Bref. A análise estatística foi realizada utilizando o programa estatístico MINITAB version16. Além de análises descritivas das variáveis categorizadas, utilizou-se o teste qui-quadrado de Person com o intuito de verificar possíveis associações entre as variáveis sociodemográficas, comportamentais, clínicas, níveis de atividade física, sintomatologia climatérica, qualidade de vida e a função sexual das mulheres estudadas. Desenvolveu-se a regressão logística para verificar a influência dessas variáveis sobre a disfunção sexual. Considerou-se o nível de significância de 5% para todos os testes. Os resultados mostraram que a média de idade das mulheres estudadas foi de 49,8 (±8,1) anos. Do total dessas mulheres, 67% apresentaram disfunção sexual. Observou-se que 54,5% delas se encontravam na pré-menopausa. Avaliando a influência das variáveis sobre a função sexual; faixa etária (56-65) (p<0,001), estado civil (divorciada/separada) (p < 0,001), escolaridade (baixa) (p=0,017), menopausa (p < 0,001), histerectomia (p = 0,016), nível de atividade física (sedentária) (p=0,002), sintomas do climatério (forte) (p<0,001) e qualidade de vida (baixa) (p<0,001), estiveram associados à disfunção sexual em mulheres de meia idade. Concluiu-se neste estudo que fatores sociodemográficos, clínicos, comportamentais, níveis de atividade física, sintomatologia climatérica e qualidade de vida influenciam significativamente a função sexual na mulher de meia idade

Sex group composition, social interaction, and metabolism in the fish Nile tilapia

O objetivo deste trabalho foi testar se grupos monossexuais de machos gastam mais energia e exibem perfil agonístico diferente de grupos formados por um macho e uma fêmea na tilápia-do-Nilo (Oreochromis niloticus). Tais diferenças são esperadas, pois machos e fêmeas competem por diferentes recursos reprodutivos. Foram utilizadas duplas de machos (MM) e duplas de macho-fêmea (MF) que permaneceram pareadas por 40 minutos. Durante esse período foi feito o registro da interação agonística (10 minutos iniciais e 10 minutos finais do pareamento) e determinado o gasto energético (consumo de O2) pelo Método de Winckler. A latência para o início dos confrontos (média ± DP, MM = 27,40 ± 25,15 s; MF = 14,22 ± 21,19 s; Mann-Whitney, U = 33,50, P = 0,21) e a freqüência de todas as unidades comportamentais (média ± DP, MM < 72,30 ± 25,29; MF < 73,50 ± 21,65.10/min; Mann-Whitney, P > 0,10) foram semelhantes entre os grupos MM e MF nos 10 minutos iniciais. Isso indica que cada intruso foi considerado um potencial competidor no início da interação. No entanto, a freqüência de ondulação (interação também exibida durante a corte) foi maior para o residente do grupo MF nos 10 minutos finais (média ± DP, MM = 3,56 ± 5,89; MF = 8,56 ± 4,00.10/min; Mann-Whitney, U = 15,50, P = 0,01). A freqüência de fuga, entretanto, foi menor para o intruso do mesmo grupo (média ± DP, MM = 3,90 ± 4,33; MF = 0,44 ± 0,96.10/min; Mann-Whitney, U = 23,50, P = 0,04). Além disso, o perfil agonístico no grupo MM foi composto por um maior número de itens comportamentais do que o MF (para residentes e intrusos). Apesar das diferenças comportamentais, o consumo de O2 não foi afetado pela composição sexual do grupo (média ± DP, MM = 1,93 ± 0,54; MF = 1,77 ± 0,46 mgO2.g peso seco-1.40/min; t-teste de Student, t = 0,71, P = 0,49).

Tempo livre de lazer e composição corporal de escolares do meio-oeste catarinense

The aim of this study was to investigate the relationship between free time for leisure and body composition of students in the crucial ninth year. (N = 228) of towns in the Midwest catarinense. We used the Adolescent Behavior Questionnaire of Santa Catarina (COMPAC) to assess lifestyle, considering that Active schoolchildren during the week, accumulated 300 or more minutes of moderate or vigorous physical activity (MVPA). Were used the time to 2 hours or more / day to determine the time of excessive use of TV and computer video game. To analyze body composition were used two criteria: the Body Mass Index (BMI) and sum of skinfolds (EDC). It was observed a proportion of 67.3% of girls and 68.7% of boys assets and more than 98% of students were using excessive TV time, computer and video game. In the classification by EDC, most of the boys showed great or low levels of body composition, while more than half of girls were classified at higher levels. As for BMI, most boys and girls had not overweight. Significant difference in the comparison of total minutes per week of MVPA reported between the groups, the second criterion of EDC and BMI for girls but not for BMI in boys. It is concluded that students with higher accumulation in minutes of MVPA showed better body composition indicators, but no significant difference was found when compared active groups with inactive, according to criteria used.

Plant or fungal sequences? An alternative optimized PCR protocol to avoid ITS (nrDNA) misamplification

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Identificação e caracterização de genes vip e cry coleóptero-específicos em isolados de Bacillus thuringiensis

O objetivo deste trabalho foi identificar e caracterizar os genes cry3, vip1, vip2 e vip1/vip2 em uma coleção de 1.078 isolados de Bacillus thuringiensis potencialmente tóxicos para larvas de coleópteros. Foram utilizados pares de oligonucleotídeos iniciadores gerais obtidos a partir de regiões conservadas dos genes e do alinhamento de sequências consenso. Posteriormente, os isolados positivos foram caracterizados por meio da técnica de PCR‑RFLP, tendo-se utilizado enzimas de restrição específicas, para identificar novas subclasses de genes nos isolados. Cento e cinquenta e um isolados foram positivos para os genes avaliados, com maior frequência para o gene vip1/vip2 (139 isolados). Pela técnica de PCR‑RFLP, foram observados 14 perfis polimórficos, o que indica a presença de diferentes alelos e, consequentemente, de distintas subclasses desses genes.

Imunorreatividade da p53 associada à ausência de mutações no gene TP53 em linfomas caninos

Sabendo-se da influência das mutações no gene TP53 no desenvolvimento das neoplasias e da discrepância entre os resultados obtidos pelas técnicas de sequenciamento e imunoistoquímica, esta pesquisa teve como objetivo relacionar a sequência do TP53 com a imunorreatividade da p53. Foram obtidas amostras de linfoma de 12 cães. O diagnóstico histopatológico foi determinado pela classificação de Kiel. O imunofenótipo e a imunomarcação da p53 foram determinados por imunoistoquímica. Para reação com a p53, utilizou-se anticorpo policlonal anti-p53 (CM1) na diluição de 1:500. A região do gene TP53 compreendida entre os exons quatro e nove foi amplificada por PCR e submetida ao sequenciamento. Apesar dos resultados obtidos pela imunoistoquímica, nenhuma mutação foi encontrada nas sequências analisadas. Conclui-se que a imunorreatividade da p53 pela imunoistoquímica não pode ser atribuída à presença de mutações no domínio central do gene TP53.

Redes sociais e a lógica do desvio comportamental em relação às normas de trabalho

This paper aim to check a hypothesis that assumes several behaviors related to social work norm´s obeying as a phenomenon that can be explained by actor´s social network structure and the rational choice processes related to the social norm inside that network, principally the payoff´s analysis received by the closest actors, or neighbors, at a social situation. Taking the sociological paradigm of rational action theory as a basis, the focus is on a debate about the logic of social norms, from Émile Durkheim´s method to Jon Elster´s theory, but also including social network analysis´s variables according to Robert Hanneman; and also Vilfredo Pareto´s constants related to human sociability, at the aim to detect elements that can help the scholars to develop an agent based model which could explain the sociological problem of deviance by a better way than the common sense´s view about morality and ethics at a social work environment

Efeitos comportamental, clínico e analgésico promovidos pela injeção epidural preventiva de morfina, xilazina ou clonidina, em equinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Uso de morfina, xilazina e meloxicam para o controle da dor pós-operatória em cadelas submetidas à ovariossalpingo-histerectomia

Foram realizados estudos empregando-se analgésicos por via epidural e subcutânea em cadelas de diferentes raças e idades, submetidas à castração mediante celiotomia. Vinte animais foram tranquilizados e anestesiados com tiletamina-zolazepam, e aleatoriamente distribuídos em quatro grupos (n=5), de acordo com o fármaco e a via de administração. Os do grupo morfina (GM) foram submetidos à anestesia epidural no espaço lombossacro, com morfina (0,1mg/kg) associada ao cloreto de sódio a 0,9%. Aos do grupo xilazina (GX), foram administrados xilazina (0,2mg/kg) e cloreto de sódio a 0,9%. Os do grupo meloxicam (GME) receberam 0,2mg/kg do anti-inflamatório meloxicam associado ao cloreto de sódio a 0,9%, injetado pela via subcutânea. Os do grupo-controle (CG) receberam apenas cloreto de sódio a 0,9%. O volume final para as injeções epidurais foi padronizado para 0,3mL/kg. A mensuração inicial da concentração de cortisol plasmático, do ritmo cardíaco, da frequência respiratória e os parâmetros comportamentais foram registrados imediatamente antes do procedimento cirúrgico (M1). Registros adicionais foram apresentados às 2, 6, 12 e 24 horas após o procedimento cirúrgico (M2, M3, M4 e M5, respectivamente). As variáveis comportamentais foram avaliadas por meio de sinais clínicos e seus respectivos escores. em GX foram observadas depressão respiratória, bradicardia e concentração de cortisol mais alta do que o registrado no GM. A analgesia obtida pelo meloxicam foi considerada ineficiente. É possível concluir que a morfina, via epidural, promoveu menor incidência de efeitos colaterais e melhor analgesia e bem-estar animal.