Frequency of gene ace i polymorphism i-d in athletes of different sports / Frequência do polimorfismo i-d do gene eca i em atletas de diferentes esportes / Frecuencia del polimorfismo i-d del gen eca i en atletas de diferentes deportes

ABSTRACT Introduction: The angiotensin-converting enzyme I-D (ACE) polymorphism gene is one of the most widely investigated genetic variations in sports science. Apparently, allele I is related to endurance sports, while allele D is related to power-strength activities. Nevertheless, studies have presented controversial results when it comes as to its occurrence in a variety of sports. Objective: This study aims to evaluate the frequency of gene ACE polymorphism I-D in professional athletes of collective or individual sports. Methods: Five mL blood were collected from 189 subjects divided into two groups: athletes (AG, n=127, wrestling, taekwondo, soccer, futsal and handball) and non-athletes (NAG, n=62). The athletes group was subdivided by group modalities, into: collective and individual. Both groups were further subdivided into male and female. Thus, we have the groups FAC= collective female, FAI= individual female, MAC= collective male, and MAI= individual male. The statistical analysis was carried out by frequency test, and the Hardy- Weinberg equilibrium by the x² test. Results: The results for the AG group indicated the following frequencies: DD=7%, ID=44% and II=49%. Allele frequency: D=29% and I=71%. For the NAG, the results were: DD=6.5%, ID=45.2% and II=48%. Allele frequency: D=29% and I=71%. The AG genotypic and allele frequencies did not differ statistically from those of the NAG (p= 0.982 and p= 0.984, respectively). However, we noticed that the genotypes II and ID frequencies were significantly higher than those of the DD. Conclusion: It can be concluded that the genotypic and allelic I-D frequencies of the ACE gene do not seem to influence performance in either group or individual sports. ACTN3 genotype frequencies did not vary significantly between male and female control subjects, and overall, there was no significant deviation from Hardy-Weinberg (H-W) equilibrium. Level of evidence I; Diagnostic studies-Investigating diagnostic test.

RESUMO Introdução: O polimorfismo I-D do gene da enzima conversora da angiotensina (ECA) é uma das variações genéticas mais amplamente investigadas na ciência do esporte. Aparentemente, o alelo I está relacionado aos esportes de resistência e o alelo D às atividades de força. Entretanto, os estudos têm apresentado resultados controversos quanto a sua ocorrência em diversos esportes. Objetivo: O presente estudo pretende avaliar a frequência do polimorfismo I-D do gene da ECA em atletas profissionais de esportes coletivos ou individuais. Métodos: Cinco mL de sangue foram coletados de 189 indivíduos divididos em dois grupos: atletas (GA, n=127, praticantes de luta livre, taekwondo, futebol, futsal ou handebol) e não atletas (GNA, n=62). O grupo de atletas foi subdividido de acordo com a modalidade: coletiva e individual. Ambos os grupos foram também subdivididos em masculino e feminino. Portanto, temos os grupos FAC = feminino coletivo, FAI = feminino individual, MAC = masculino coletivo, MAI = masculino individual. A análise estatística foi realizada através do teste de frequência e o equilíbrio de Hardy-Weinberg pelo teste x². Resultados: Os resultados para o GA indicaram as seguintes frequências: DD=7%, ID=44% e II=49%. Frequência alélica: D=29% e I=71%. Para o GNA, os resultados foram: DD=6,5%, ID=45,2% e II=48%. Frequência alélica: D=29% e I=71%. As frequências genotípicas e alélicas do GA não se diferiram estatisticamente daquelas do GNA (p= 0,982 e p= 0,984, respectivamente). Entretanto, notamos que as frequências dos genótipos II e ID se apresentaram significativamente maiores do que aquelas do DD. Conclusão: Pode-se concluir que as frequências I-D genotípicas e alélicas do gene da ECA não pareceram influenciar o desempenho tanto nos esportes individuais como coletivos. As frequências do genótipo ACTN3 não variaram significativamente entre os indivíduos de controle de ambos os sexos, e, no geral, não houve um desvio significativo do equilíbrio de Hardy-Weinberg (H-W). Nível de evidência I; Estudos diagnósticos-Investigação de um exame para diagnóstico.

RESUMEN Introducción: El polimorfismo I-D del gen de la enzima convertidora de la angiotensina (ECA) es una de las variaciones genéticas más ampliamente investigadas en la ciencia del deporte. Aparentemente, el alelo I está relacionado a los deportes de resistencia y el alelo D a las actividades de fuerza. Entretanto, los estudios han presentado resultados controvertidos cuanto a su ocurrencia en diversos deportes. Objetivo: El presente estudio pretende evaluar la frecuencia del polimorfismo I-D del gen de la ECA en atletas profesionales de deportes colectivos o individuales. Métodos: Cinco mL de sangre fueron recolectados de 189 individuos divididos en dos grupos: atletas (GA, n=127 practicantes de lucha libre, taekwondo, fútbol, futsal y hándbol) y no atletas (GNA, n=62). El grupo de atletas fue subdividido de acuerdo con la modalidad: colectiva e individual. Ambos grupos fueron también subdivididos en masculino y femenino: Por lo tanto, tenemos los grupos FAC= colectivo femenino, FAI= femenino individual, MAC= masculino colectivo, MAI= masculino individual. El análisis estadístico fue realizado a través del test de frecuencia, y el equilibrio de Hardy-Weinberg por el test x². Resultados: Los resultados de GA indicaran las siguientes frecuencias: DD=7%, ID=44% y II=49%. Frecuencia alélica: D=29% e I=71%. Para el GNA, los resultados fueron: DD=6,5%, ID=45,2% y II=48%. Frecuencia alélica: D=29% e I=71%. Las frecuencias genotípicas y alélicas del GA no difirieron estadísticamente de las del GNA (p=0,982 y p=0,984, respectivamente). Entretanto, notamos que las frecuencias de los genotipos II e ID se presentaron significativamente mayores que aquellas del DD. Conclusión: Se puede concluir que las frecuencias I-D genotípicas y alélicas del gen de la ECA no parecieron influenciar el desempeño tanto en los deportes individuales como colectivos. Las frecuencias del genotipo ACTN3 no variaron significativamente entre los individuos de control de ambos sexos y, en general, no hubo desvío significativo del equilibrio de Hardy-Weinberg (H-W). Nivel de evidencia I; Estudios diagnósticos-Investigación de un examen para diagnóstico.

Production of thermostable ß-glucosidase and CMCase by Penicillium sp: LMI01 isolated from the Amazon region

Background: Cellulolytic enzymes of microbial origin have great industrial importance because of their wide application in various industrial sectors. Fungi are considered the most efficient producers of these enzymes. Bioprospecting survey to identify fungal sources of biomass-hydrolyzing enzymes from a high-diversity environment is an important approach to discover interesting strains for bioprocess uses. In this study, we evaluated the production of endoglucanase (CMCase) and ß-glucosidase, enzymes from the lignocellulolytic complex, produced by a native fungus. Penicillium sp. LMI01 was isolated from decaying plant material in the Amazon region, and its performance was compared with that of the standard isolate Trichoderma reesei QM9414 under submerged fermentation conditions. Results: The effectiveness of LMI01 was similar to that of QM9414 in volumetric enzyme activity (U/mL); however, the specific enzyme activity (U/mg) of the former was higher, corresponding to 24.170 U/mg of CMCase and 1.345 U/mg of ß-glucosidase. The enzymes produced by LMI01 had the following physicochemical properties: CMCase activity was optimal at pH 4.2 and the ß-glucosidase activity was optimal at pH 6.0. Both CMCase and ß-glucosidase had an optimum temperature at 60°C and were thermostable between 50 and 60°C. The electrophoretic profile of the proteins secreted by LMI01 indicated that this isolate produced at least two enzymes with CMCase activity, with approximate molecular masses of 50 and 35 kDa, and ß-glucosidases with molecular masses between 70 and 100 kDa. Conclusions: The effectiveness and characteristics of these enzymes indicate that LMI01 can be an alternative for the hydrolysis of lignocellulosic materials and should be tested in commercial formulations.

Assessment of petroleum biodegradation for Bacillus toyonensis by the using redox indicator 2,6 dichlorophenol indophenol / Avaliação da biodegradação do petróleo por Bacillus toyonensis, usando o indicador redox 2,6 diclorofenol indofenol

Petroleum degrading microorganisms have been isolated from different environments with the purpose of being used in bioremediation processes in areas impacted by petroleum spills. The objective of this study was to evaluate the ability of Bacillus toyonensis AM07 strain to metabolize petroleum compounds. The strain was isolated from the effluent dike of the Urucu Petroleum Province, Coari - Amazonas, Brazil. The degrading activity of B. toyonensis was evaluated by the colorimetric method using the redox indicator 2,6-dichlorophenol indophenol (DCPIP). Thus, the microorganism was inoculated into minimal medium with DCPIP, and with petroleum as the sole carbon source. The degradation potential of the microorganism was found by changing the DCPIP staining and absorbance readings 600nm. The results obtained demonstrated that the bacterial strain was able to degrade petroleum by altering the color of the medium from blue to colorless and by reducing the concentration of the indicator in the absorbance readings. B. toyonensis AM07 strain has shown good performance in the petroleum degradation assays and may be used in the future in remediation technologies for hydrocarbon impacted environments.

Microrganismos degradadores de petróleo têm sido isolados de diferentes ambientes com a finalidade de serem utilizados em processos de biorremediação de áreas impactadas com derrames de petróleo. O objetivo deste estudo foi avaliar a capacidade da linhagem de Bacillus toyonensis AM07, isolada do dique de efluente da Província Petrolífera de Urucu, Coari - Amazonas, Brasil, em metabolizar compostos do petróleo. A atividade degradadora do B. toyonensis foi avaliada pelo método colorimétrico, utilizando indicador redox 2,6-diclorofenol indofenol (DCPIP). Assim, o microrganismo foi inoculado em meio mínimo com DCPIP e petróleo como única fonte de carbono. O potencial de degradação do microrganismo foi constatado mediante a mudança de coloração DCPIP e leituras de absorbância 600nm. Os resultados obtidos demonstraram que a cepa bacteriana foi capaz de degradar petróleo, alterando a coloração do meio de azul para incolor e reduzindo a concentração do indicador nas leituras de absorbâncias. A cepa de B. toyonensis AM07 mostrou bom desempenho nos ensaios de degradação do petróleo, podendo ser utilizada, no futuro, em tecnologias de remediação de ambientes impactados por hidrocarbonetos.

Petroleum biodegrading and co-resistance to antibiotics by Serratia marcescens strain isolated in Coari, Amazonas / Biodegradação do petróleo e corresistência a antibióticos por Serratia marcescens isolada em Coari, Amazonas

Serratia marcescens is a Gram-negative bacillus, anaerobic facultative belonging to the family Enterobacteriaceae. S. marcescens strains are able to grow in the presence of different xenobiotic compounds, among them, petroleum and heavy metals. Xenobiotic resistant strains develop concomitant resistance to multiple antibiotics, referred to as co-resistance. The AMS212 strain was submitted to the microplate qualitative DCPIP - redox 2,6 dichlorophenol indophenol method. The quantitative test was carried out in Erlenmeyer flasks, followed by the change of color with the absorbance readings, trough the colorimetric method. The antibiotic resistance profile was evaluated by the Kirby -Bauer method. In the qualitative assay, the AMS212 strain altered the color of the DCPIP, which changed from blue to colorless, confirming that petroleum biodegradation occurred. In the quantitative test, the readings were decreasing, confirming that the concentration of DCPIP decreased as a function of the incubation time. The susceptibility test revealed that the AMS212 strain presented multiresistance to four different antibiotics. S. marcescens presented high performance in the biodegradation of petroleum, opening possibility to use it in projects involving the remediation of impacted areas. The expression of the antibiotic co-resistance phenotype confirms that the AMS212 strain is able to withstand different environmental aggressions.

Serratia marcescens é um bacilo Gram-negativo, anaeróbio facultativo, pertencente à família Enterobacteriaceae. Linhagens de S. marcescens são capazes de crescer na presença de diferentes compostos xenobióticos, dentre eles, petróleo e metais pesados. Linhagens resistentes a xenobióticos desenvolvem concomitante resistência a múltiplos antibióticos, denominada corresistência. A linhagem AMS212 foi submetida ao método colorimétrico com indicador DCPIP - redox 2,6 diclorofenol indofenol, qualitativo, em microplacas. O teste quantitativo foi realizado em frascos Erlenmeyer, acompanhando-se a mudança de coloração, com as leituras das absorbâncias. Avaliou-se o perfil de resistência a antibióticos pelo método de Kirby-Bauer. No ensaio qualitativo, a linhagem AMS212 alterou a cor do DCPIP, que passou de azul para incolor, confirmando que ocorreu biodegradação do petróleo. No teste quantitativo, as leituras foram decrescentes, confirmando que a concentração do DCPIP diminuiu em função do tempo de incubação. O teste de susceptibilidade revelou que a linhagem AMS212 apresenta multirresistência a quatro antibióticos diferentes. S. marcescens apresentou alto desempenho na biodegradação do petróleo, abrindo possibilidade de utilizá-la em projetos envolvendo a remediação de áreas impactadas. A expressão do fenótipo de corresistência a antibióticos confirma que a linhagem AMS212 é capaz de resistir a diferentes agressões ambientais.

Xylitol production and furfural consumption by a wild type Geotrichum sp

Background: Xylitol is a five carbons polyol with promising medical applications. It can be obtained from chemical D-xylose reduction or by microbial fermentation of Sugarcane Bagasse Hemicellulosic Hydrolysate. For this last process, some microbial inhibitors, as furfural, constitute severe bottleneck. In this case, the use of strains able to produce xylitol simultaneously to furfural neutralization is an interesting alternative. A wild-type strain of Geotrichum sp. was detected with this ability, and its performance in xylitol production and furfural consumption was evaluated. Furthermore, were analyzed its degradation products. Results: Geotrichum sp. produced xylitol from D-xylose fermentation with a yield of 0.44 g-g-1. Furfural was fully consumed in fermentation assay and when provided in the medium until concentration of 6 g-L-1. The furfural degradation product is not an identified molecule, presenting a molecular weight of 161 g-mol-1, an uncommon feature for the microbial metabolism of this product. Conclusion: This strain presents most remarkable potential in performing furfural consumption simultaneous to xylitol production. Subsequent efforts must be employed to establish bioprocess to simultaneous detoxification and xylitol production by Geotrichum sp.

Isolation, taxonomic identification and investigation of the biotechnological potential of wild-type Meyerozyma guilliermondii associated with amazonian termites able to ferment D-xylose / Isolamento, identificação taxonômica e investigação do potencial biotecnológico de linhagem selvagem de Meyerozyma guilliermondii fermentadora de d-xilose associada a cupins da Amazônia

A wild-type yeast that could ferment D-xylose was isolated from the abdominal content of Nasutitermes sp. collected in the Central Amazon rainforest using sugarcane bagasse hemicellulosic hydrolyzate (SBHH) as selective medium. The yeast was identified as Meyerozyma guilliermondii. Its ability to ferment D-xylose was assessed using liquid medium containing Durham tubes. A fermentometer assay showed a low ethanol yield using D-xylose as the carbon source. Cell viability after heat shock and ethanol shock was 39.8% and 56.0%, respectively. Cultivation in SBHH (pH = 5.0) showed its capability to perform saccharification of this substrate, increasing total reducing sugar concentration to 42.6%. The log phase was observed between 36 and 108 hours of cultivation with a highest specific growth rate (µMAX) of 0.10 h-1. After 120 hours, 79.5% of total reducing sugar was consumed giving a biomass yield of 0.52 g/g. The final pH of SBHH (7.6) showed that M. guilliermondii was able to neutralize the acids of this substrate. These results agree with some predictions in the early eighties, which stated that investigations about microbial content of termite guts would provide new tools for bioconversion of lignocellulosic biomass to fuels and other added-value chemicals. This work is the first report for this species associated with termites in the Amazonian habitat.

Uma levedura selvagem fermentadora de D-xilose foi isolada do conteúdo abdominal de Nasutitermes sp., coletado na Amazônia Central usando Hidrolisado Hemicelulósico de Bagaço de Cana-de-açúcar (HHCA) como meio seletivo. A levedura foi identificada como Meyerozyma guilliermondii. Sua capacidade de fermentar D-xilose foi avaliada usando meio líquido contendo tubos de Durham. O isolado demonstrou moderada tolerância ao calor e ao etanol, com viabilidade celular de 39,8% e 56,0%, respectivamente, após submetida a estes fatores limitantes. O ensaio em fermentômetro demonstrou baixo rendimento de etanol usando D-xilose como fonte de carbono. O cultivo em HHCA (pH = 5,0) demonstrou sua capacidade de executar sacarificação e neutralização deste substrato, com aumento da concentração de açúcar redutor total em 42,6% e elevação do pH para 7,6. A fase log foi observada entre 36 e 108 horas de cultivo, com máxima taxa de crescimento específico (µMAX) de 0,10 h-1. Depois de 120 horas, 79,5% do açúcar redutor total foi consumido, com rendimento de biomassa de 0,52 g/g. Estes resultados endossam as predições de alguns autores, os quais propuseram, no início dos anos 80, que a investigação da microbiota intestinal de cupins proveria novas ferramentas para utilização de biomassa lignocelulósica e seus derivados. Este trabalho é o primeiro a reportar a ocorrência de Meyerozyma guilliermondii associada a cupins da Amazônia Central.

Chlamydia trachomatis infection in a sample of northern Brazilian pregnant women: prevalence and prenatal importance

There are limited data regarding prevalence of Chlamydia trachomatis infection among northern Brazilian pregnant women. OBJECTIVE: The purpose of this study was to estimate the prevalence of chlamydial infection among pregnant women in their third trimester and to determine the repercussion of this infection on their offspring. METHODS: In the first phase of this study 100 pregnant women receiving prenatal care in a local public university hospital were examined to assess the prevalence of genital C. trachomatis infection by polymerase chain reaction technique. In the second phase, 88 pregnant women were prospectively evaluated for premature rupture of membranes, puerperal consequences associated with chlamydial infection, and neonates were checked for low-birth weight. RESULTS: The prevalence rate of chlamydial infection was 11%, and 72.7% of the positive participants were predominantly less than 30 years of age (p = 0.1319). A total of 36.4% of the participants had premature rupture of membranes (p = 0.9998). Neither low-birth weight infants nor preterm delivery were observed. A cohort of 16 newborn babies were followedup up to 60 days of life to ascertain outcome: 50% had respiratory symptoms. Neonates born to infected mothers had a higher risk to develop respiratory symptoms in the first 60 days of life. CONCLUSION: The scarcity of data about the effects of chlamydial infection on pregnancy and neonatal outcomes justified this study. Diagnosing and treating chlamydial infection during the third trimester of pregnancy may prevent neonate infection. Therefore, preventive screening should be seen as a priority for early detection of asymptomatic C. trachomatis infection as part of local public health strategies.

Cultivation of Trichosporon mycotoxinivorans in sugarcane bagasse hemicellulosic hydrolyzate

Background: The yeast strain IB09 was isolated from the gut of Calosoma sp. (Carabidae, Coleoptera, Insecta) that were collected in the central Amazon rainforest. First, tolerance of the strain to ethanol and heat was tested. Then, IB09 was cultivated in a medium using sugarcane bagasse hemicellulosic hydrolyzate as a carbon source, and cell growth (OD600), specific growth rate (uMAX, h-1), biomass yield (Y B, g.g-1) and relative sugar consumption (RSC, percent) were evaluated. Taxonomic identification was determined by sequencing the ITS1 region of IB09 and comparing it to sequences obtained from the GenBank database (NCBI). Results: IB09 showed both ethanol tolerance and thermotolerance. Relative sugar consumption indicated that IB09 was able to perform saccharification of sugarcane bagasse hemicellulosic hydrolyzate, increasing the total reducing sugar concentration by approximately 50 percent. The μMAX value obtained was 0,20, indicating that cell growth was slow under the assessed conditions. Biomass yield was 0,701 g per g of consumed sugar, which is relatively high when compared with other findings in the literature. After 120 hrs of cultivation, 80,1 percent of total reducing sugar had been consumed. Sequencing of the ITS1 region identified IB09 as Trichosporon mycotoxinivorans. Conclusion: This is the first report to document this species in the central Amazon rainforest at this host. Trichosporon mycotoxinivorans has great biotechnological potential for use in the saccharification of sugarcane bagasse hemicellulosic hydrolyzate and for biomass production with this substrate as carbon source.

Bacterial diversity in the saliva of patients with different oral hygiene indexes

The objective of the present study was to evaluate the bacterial diversity in the saliva of patients with different oral hygiene indexes using of two 16S rRNA gene libraries. Each library was composed of samples from patients with different averages of the differentiated Silness-Löe biofilm index: the first library (A) with an index between 1.0 and 3.0 (considered a high index) and the second library (B) between 0 and 0.5 (considered a low index). Saliva DNA was extracted and the 16S rRNA gene was amplified and cloned. The obtained sequences were compared with those stored at NCBI and RDP GenBank. The saliva of patients with high index presented five known genera - Streptococcus, Granulicatella, Gemella, Veillonella and Peptostreptococcus - and 33.3% of nonculturable bacteria grouped into 23 operational taxonomic units (OTUs). The saliva of patients with low index differed significantly from the first library (p=0.000) and was composed of 42 OTUs distributed into 11 known genera - Streptococcus, Granulicatella, Gemella, Veillonella, Oribacterium, Haemophilus, Escherichia, Neisseria, Prevotella, Capnocytophaga, Actinomyces - including 24.87% of nonculturable bacteria. It was possible to conclude that there is greater bacterial diversity in the saliva of patients with low dental plaque in relation to patients with high dental plaque.

O objetivo do presente estudo foi avaliar a diversidade bacteriana da saliva de pacientes com diferentes índices de higiene bucal através da construção de duas bibliotecas do gene 16S rRNA. Cada biblioteca foi composta por amostras de saliva de pacientes com índice de biofilme dental de Silness-Löe diferenciado, sendo a primeira (A) com índice de 1,0 a 3,0 (denominada de alto índice) e a segunda (B), entre 0 a 0,5 (denominada de baixo índice). O DNA da saliva foi extraído e o gene 16S rRNA foi amplificado, clonado e sequenciado. As sequências obtidas foram comparadas com aquelas armazenadas no GenBank do NCBI e RDP. A saliva de pacientes com alto índice de biofilme dental apresentou cinco gêneros conhecidos: Streptococcus, Granulicatella, Gemella, Veillonella e Peptostreptococcus e 33,3% de bactérias não-cultivadas, agrupados em 23 unidades taxonômicas operacionais (UTOs). A saliva de pacientes com baixo índice de biofilme dental, foi diferente significativamente da primeira (p=0,000) e foi composta de 42 UTOs, distribuídas em 11 gêneros conhecidos: Streptococcus, Granulicatella, Gemella, Veillonella, Oribacterium, Haemophilus, Escherichia, Neisseria, Prevotella, Capnocytophaga, Actinomyces, além de 24,87% de bactérias não-cultivadas. Pode-se concluir que existe maior diversidade bacteriana na saliva de pacientes com baixo índice de biofilme dental em relação a pacientes com alto índice de biofilme dental.

Anticoagulant activity in salivary gland homogenates of Thyrsopelma guianense (Diptera: Simuliidae), the primary vector of onchocerciasis in the Brazilian Amazon

In this study, anticoagulant activity was detected in salivary gland homogenates (SGHs) of Thyrsopelma guianense (Diptera: Simuliidae). The SGH yielded 1.07 ìg ± 0.03 (n = 15) of total soluble protein per pair of glands. In addition, following SDS-PAGE (12.5 percent gel) and silver nitrate staining, 12 polypeptides with molecular weights ranging from 14-69 kDa were detected in all physiological ages analyzed (12 h, 24 h, 48 h and 72 h following emergence). Coagulation bioassays showed that the SGHs had activities that interacted at all levels of coagulation (the intrinsic, extrinsic and common pathways), by extending the plasma recalcification time, prothrombin time, thrombin time. This is the first report on the activity of salivary gland proteins from the main vector of onchocerciasis in Brazil. We also suggest detailed studies on the morphology and function of the salivary glands in order to understand the role of these proteins in host/vector interactions.

Isolation and characterization of BliAI, an isoschizomer of ClaI from Bacillus licheniformis

The restriction endonuclease BliAI, an isoschizomer of ClaI, which recognizes the sequence 5'- AT¼CGAT - 3', was purified from a natural isolate identified as Bacillus licheniformis. The restriction endonuclease was isolated from cell extracts using single-step purification by phosphocellulose column chromatography. The restriction endonuclease is active at 37°C and over a wide range of pH and salt concentration. The molecular weight of the purified restriction enzyme is consistent with a value of 39 kDa.

A endonuclease de restrição BliAI, um isoesquisômero de ClaI, que reconhece a seqüência 5'- AT¼CGAT - 3', foi purificada de um isolado natural identificado como Bacillus licheniformis. A endonuclease de restrição em questão foi isolada a partir de um extrato celular em um único passo cromatográfico utilizando uma coluna contendo a resina fosfocelulose. A endonuclease de restrição é ativa à 37°C e em uma ampla escala de pH e concentração de sais. O peso molecular da enzima purificada corresponde a um valor de kDa 39.

BpuAmI: a novel SacI neoschizomer from Bacillus pumilus discovered in an isolate from Amazon Basin, recognizing 5'-GAG FONT FACE=Symbol>¯ /FONT>CTC-3'

A strain of Bacillus pumilus was isolated and identified from water samples collected from a small affluent of the Amazon River. Type II restriction endonuclease activity was detected in these bacteria. The enzyme was purified and the molecular weight of the native protein estimated by gel filtration and SDS-PAGE. The optimum pH, temperature and salt requirements were determined. Quality control assays showed the complete absence of "nonspecific nucleases." Restriction cleavage analysis and DNA sequencing of restriction fragments allowed the unequivocal demonstration of 5´GAG FONT FACE=Symbol>¯ /FONT>CTC3´ as the recognition sequence. This enzyme was named BpuAmI and is apparently a neoschizomer of the prototype restriction endonuclease SacI. This is the first report of an isoschizomer and/or neoschizomer of the prototype SacI identified in the genus Bacillus.

Uma linhagem de Bacillus pumilus foi isolada e identificada de amostras de águas coletadas em um pequeno Igarapé do Rio Amazonas. Foi detectada atividade de restrição do tipo II nesta bactéria. A enzima foi purificada e o peso molecular da proteína nativa foi estimado por gel filtração e por eletroforese em gel de poliacrilamida. Foram determinados, o pH e temperatura ótimos e as necessidades de sais. Os ensaios do controle de qualidade mostraram uma ausência completa de "nucleases não específicas". As analises das clivagens e o seqüenciamento do DNA dos fragmentos de restrição permitiram uma demonstração inequívoca de que 5´GAG FONT FACE=Symbol>¯ /FONT>CTC 3´ é a seqüência de reconhecimento da enzima. Esta enzima foi denominada de BpuAmI e aparentemente é um neoesquisômero da enzima protótipo SacI. Este é o primeiro relato de um isoesquisômero e/ou neoesquisômero da enzima protótipo SacI identificada no gênero Bacillus.

BpuAmI: a novel SacI neoschizomer from Bacillus pumilus discovered in an isolate from Amazon Basin, recognizing 5'-GAGü®CTC-3'

Uma linhagem de Bacillus pumilus foi isolada e identificada de amostras de águas coletadas em um pequeno Igarapé do Rio Amazonas. Foi detectada atividade de restrição do tipo II nesta bactéria. A enzima foi purificada e o peso molecular da proteína nativa foi estimado por gel filtração e por eletroforese em gel de poliacrilamida. Foram determinados, o pH e temperatura ótimos e as necessidades de sais. Os ensaios do controle de qualidade mostraram uma ausência completa de "nucleases não específicas". As analises das clivagens e o seqüenciamento do DNA dos fragmentos de restrição permitiram uma demonstração inequívoca de que 5üLGAG¼CTC 3üL é a seqüência de reconhecimento da enzima. Esta enzima foi denominada de BpuAmI e aparentemente é um neoesquisômero da enzima protótipo SacI. Este é o primeiro relato de um isoesquisômero e/ou neoesquisômero da enzima protótipo SacI identificada no gênero Bacillus.

Oral focal epithelial hyperplasia: report of five cases

Hiperplasia epitelial focal ou doença de Heck é uma rara doença contagiosa causada pelos papilomavírus tipo 13 e 32 que foi inicialmente descrita entre populações nativas americanas. Esta doença caracteriza-se pela ocorrência de pequenas e múltiplas pápulas ou nódulos na cavidade bucal, especialmente nos lábios, mucosa bucal e língua. Este artigo descreve o diagnóstico da hiperplasia epitelial focal em cinco indígenas da Amazônia Central que procuraram tratamento na Fundação de Medicina Tropical do Amazonas (FMT-AM) utilizando critérios clínicos, reação em cadeia de polimerase (PCR) e sequenciamento de DNA.

The use of biodiversity as source of new chemical entities against defined molecular targets for treatment of malaria, tuberculosis, and T-cell mediated diseases: a review

The modern approach to the development of new chemical entities against complex diseases, especially the neglected endemic diseases such as tuberculosis and malaria, is based on the use of defined molecular targets. Among the advantages, this approach allows (i) the search and identification of lead compounds with defined molecular mechanisms against a defined target (e.g. enzymes from defined pathways), (ii) the analysis of a great number of compounds with a favorable cost/benefit ratio, (iii) the development even in the initial stages of compounds with selective toxicity (the fundamental principle of chemotherapy), (iv) the evaluation of plant extracts as well as of pure substances. The current use of such technology, unfortunately, is concentrated in developed countries, especially in the big pharma. This fact contributes in a significant way to hamper the development of innovative new compounds to treat neglected diseases. The large biodiversity within the territory of Brazil puts the country in a strategic position to develop the rational and sustained exploration of new metabolites of therapeutic value. The extension of the country covers a wide range of climates, soil types, and altitudes, providing a unique set of selective pressures for the adaptation of plant life in these scenarios. Chemical diversity is also driven by these forces, in an attempt to best fit the plant communities to the particular abiotic stresses, fauna, and microbes that co-exist with them. Certain areas of vegetation (Amazonian Forest, Atlantic Forest, Araucaria Forest, Cerrado-Brazilian Savanna, and Caatinga) are rich in species and types of environments to be used to search for natural compounds active against tuberculosis, malaria, and chronic-degenerative diseases. The present review describes some strategies to search for natural compounds, whose choice can be based on ethnobotanical and chemotaxonomical studies, and screen for their ability to bind to immobilized drug targets and to inhibit their activities. Molecular cloning, gene knockout, protein expression and purification, N-terminal sequencing, and mass spectrometry are the methods of choice to provide homogeneous drug targets for immobilization by optimized chemical reactions...

Atividade antimicrobiana de fungos endofíticos isolados de plantas tóxicas da amazônia: Palicourea longiflora (aubl. ) rich e Strychnos cogens bentham / Antimicrobial activity of endophytic fungi isolated from amazonian toxic plants: Palicourea longiflora (aubl. ) rich and Strychnos cogens bentham

Das plantas tóxicas da Amazônia Palicourea longiflora e Strychnos cogens foram isolados 571 fungos endofíticos e 74 bactérias endofíticas. Palicourea longiflora (Rubiaceae) e outras espécies desse gênero estão relacionadas a 90 por cento das mortes de gado na região Amazônica. Strychnos cogens (Loganiaceae) e outras espécies de Strychnos são utilizadas pelos indígenas na confecção de "curares". Entre os endófitos isolados de P. longiflora foram identificados os fungos: Colletotrichum sp. e seu telemorfo Glomerella sp., Guignardia sp., Aspergillus niger, Phomopsis sp. e Xylaria sp., além da bactéria Burkholderia gladioli, pertencente a um grupo de fixadoras de nitrogênio. Dos isolados de S. cogens foram identificados os fungos: Colletotrichum sp., Guignardia sp., Aspergillus niger e Trichoderma sp. Uma amostra de 79 isolados fúngicos teve seus metabólitos extracelulares bioensaiados contra microrganismos patogênicos e fitopatogênicos: 19 isolados inibiram um ou mais microrganismos-teste. Dos oito isolados com melhores resultados de inibição, as móleculas bioativas eram menores que 12.000 daltons, fato verificado pela diálise dos metabólitos.

Detection of Chlamydia trachomatis in endocervical smears of sexually active women in Manaus-AM, Brazil, by PCR

Chlamydia trachomatis is now one of the most prevalent bacteria found in classic sexually transmissible diseases (STD), and as such, constitutes a serious public health problem. We examined the prevalence of Chlamydia trachomatis, by polymerase chain reaction (PCR), in 121 sexually active women who sought treatment for STD in the Alfredo da Matta Institute of Dermatology and Venerology and the Institute of Tropical Medicine of Amazonas in Manaus, Brazil. These women were examined by a specific PCR for the chlamydial plasmid, and the nature of the amplicon was determined by restriction analysis and DNA sequencing. The PCR diagnosis revealed a prevalence of 20.7 percent infected women

Discriminação de raças primitivas de Pupunha (Bactris Gasipaes) na Amazônia brasileira por meio de marcadores moleculares (RAPDs)

The pejibaye (Bactris gasipaes Kunth, Palmae) was domesticated for it fruits by the first peoples of western Amazonia. Consequently it exhibits a landrace complex that has been partially characterized morphologically and mapped. Along the Amazonas and Solimões Rivers, in Brazil, three landraces have been proposed [Pará (Amazonas River), Solimões (lower and middle Solimões River), Putumayo (upper Solimões River)], with indications that the Solimões landrace could be an artifact of the morphometric analysis. RAPD markers were used to evaluate the three landrace hypothesis. DNA was extracted from 30 plants of each landrace maintained in the Pejibaye germplasm bank, Manaus, AM, Brazil. During PCR amplification, 8 primers generated 80 markers, Jaccard similarities were estimated, the plants were grouped with UPGMA. The dendrogram contained 2 large groups that joined at a similarity of 0.535: the group of the Pará landrace contained 26 plants of this race, 5 of the Putumayo and 1 of the Solimões; the group of the Solimões River contained 29 plants of the Solimões race, 19 of the Putumayo and 1 of the Pará. The structure of the second group suggested that there is only one landrace along the Solimões River, since the plants were mixed in sub-groups without apparent order. This marker-based genetic analysis did not support the three landrace hypothesis and suggests that the Putumayo landrace extends along the Solimões River to central Amazonia. Genetic and morphological data must now be used to evaluate this new hypothesis.

A pupunha (Bactris gasipaes Kunth, Palmae) foi domesticada por seu fruto pelos primeiros povos da Amazônia Ocidental, possuindo um complexo de raças primitivas (landraces) parcialmente caracterizado e mapeado morfologicamente. Ao longo dos Rios Amazonas e Solimões, no Brasil, foram propostas três raças primitivas [Pará (Rio Amazonas), Solimões (baixo e médio Rio Solimões), Putumayo (alto Rio Solimões)], com indicações de que a raça Solimões poderia ser artefato de análise morfométrica. Marcadores RAPDs foram usados para avaliar a hipótese de três raças. Extraiu-se DNA de 30 plantas de cada raça mantida no BAG de Pupunha em Manaus, AM, Brasil. Na amplificação por PCR, 8 primers geraram 80 marcadores, cujas similaridades de Jaccard foram estimadas para agrupamento das plantas com UPGMA. O dendrograma conteve 2 grandes grupos que juntaram-se a uma similaridade de 0,535: o grupo da raça Pará conteve 26 plantas dessa raça, 5 da Putumayo e 1 da Solimões; o grupo do Rio Solimões conteve 29 plantas da raça Solimões, 19 da Putumayo e 1 da Pará. A estrutura do segundo grupo sugere que existe apenas uma raça ao longo do Rio Solimões, pois as plantas amostradas são misturadas em sub-grupos sem ordem aparente. A análise genética não apoia a hipótese de três raças e sugere que a raça Putumayo estende-se ao longo do Rio Solimões até Amazônia central. Será necessário juntar dados genéticos com morfológicos para avaliar esta nova hipótese com mais precisão.

Expression of heretologous amylases in the yeast Saccharomyces cerevisiae

The major microbial alcoholic fermenter, the yeast Saccharomyces cerevisiae, is unable to utilize starch for ethanol production because it lacks starch degrading enzymes. Yeast genetic engineering makes it possible to obtain strains expressing amylases from a variety of different organisms. Here we present a review of the main results obtained in our group pursuing a stable recombinant S. cerevisiae strain possessing all of the enzymatic activities needed for the production of ethanol from starchy raw materials.

Clonagem do operon de resistência ao íon mercúrio do plasmidio pBH100 em escherichia coli 5K, usando o vetor pAT 153 / Cloning of the mercuric ion-resistance operon of pBH100 into escherichia coli 5K using pAT153 as vector

Resumo: Inicialmente, o plasmidio pBH100 (95 Kb) que codifica resistência, canamicina, cloranfenicol, estreptomicina e mercúrio inorgânico, foi transferido, por conjugaçäo, da linhagem selvagem Escherichia coli BH100 para a receptora. E. coli 5K. Para clonar o operon Hg plasmideos recombinantes foram construidos in vitro a partira ligaçäo (ligase) de fragmentos de pBH100, contendo os marcadores de resistência ao mercúrio, com o veiculo de clonagem pAT153, ambos digeridos coma endonuclease BamH I. Após transformaçäo de Escherihia coli 5K análises genéticas e eletroforética permitiram a detecçäo dos plasmídios recombinantes pATHg 1, pATHg2, pATHg3. Análise de restriçäo do plasmídio pATHg1, digerido com BamHI e Hind III, evidenciou três bandas de 24, 4 e 1 Kb e duas bandas de 21 e 8 Kb, respectivamente. O pATHg3 apresentou menor tamanho (12 Kb) e menor estabilidade 37 (por cento)(au)