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.

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.

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.

We evaluated the biomass of twenty Amazonian fungal isolates as a potential source of mycelium-bound lipases with hydrolytic, synthetic or enantioselective activity for biocatalysis application. We compared the hydrolytic activity of three biomass treatments (delipidated, non-delipidated and cultivated in medium without inducer). Delipidated biomass showed better results in the hydrolysis of p-nitrophenyl palmitate compared to the other two treatments for fifteen isolates. Delipidated biomass of six Aspergillus strains and UEA_115 strain showed a high synthesis activity of ethyl palmitate by transesterification in organic medium. Results were confirmed by spectrophotometry (410 nm) and gas chromatography. In this reaction, the isolate DPUA_1539 A. flavo-furcatis reached a maximum value of 668.5 ± 23.5 mU g-1. Enantioselective activity assays indicated that biomass-bound lipases from UEA_115 isolate (E = 3.58; ees = 7 ± 0) and in particular from the DPUA_1539 A. flavo-furcatis isolate (E = 24.15; ees = 91 ± 1) have the ability to discriminate enantiomers of the drug ibuprofen by ester synthesis, preferably with (R)-enantiomer. These results encourage further investigations of these fungi as potential lipase suppliers for biocatalytic processes such as biodiesel production and enantiopure drugs.

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.

O Banco Ativo de Germoplasma de Pupunha tem abundante diversidade genética em seu acervo. Por ser uma coleção viva, os recursos financeiros para caracterizar e avaliar são escassos, restringindo seu uso. Uma forma de promover o uso eficiente é criar uma coleção nuclear, um conjunto de acessos com pelo menos 70% da diversidade genética da coleção inteira, com repetição mínima. As informações geográfica, molecular e morfométricas foram sistematizadas e as populações silvestres e domesticadas estratificadas. A coleção nuclear é constituída por 10% de toda a coleção: 31 amostras de raças primitivas, cinco amostras de populações não-designadas e quatro amostras de populações silvestres. A coleção tem 92% de polimorfismo molecular e 95% da heterozigosidade do banco inteiro, com um mínimo de divergência entre elas por variância molecular e já está recebendo prioridade em manutenção, o que irá contribuir para a conservação em longo prazo

The Peach palm Active Germplasm Bank has abundant genetic diversity in its holdings. Because it is a live collection, maintenance, characterization and evaluation are expensive, restricting its use. One way to promote more efficient use is to create a Core Collection, a set of accessions with at least 70% of the genetic diversity of the full collection with minimal repetition. The available geographic, molecular marker (RAPD) and morphometric information was systematized and the populations were stratified into wild and domesticated. The Core Collection consists of 10% of the entire collection: 31 accessions of landraces, 5 accessions of non-designated populations and 4 accessions of wild populations. The Core has 92% of the molecular polymorphism and 95% of the heterozygosity of the full collection, with minimal divergence between them by molecular variance. The Core is already receiving priority maintenance, which will contribute to long term conservation.

Peach palm (Bactris gasipaes) is cultivated by many indigenous and traditional communities from Amazonia to Central America for its edible fruits, and is currently important for its heart-of-palm. The objective of this study was to investigate the mating system of peach palm, as this is important for conservation and breeding. Eight microsatellite loci were used to genotype 24 open-pollinated progenies from three populations of the Pampa Hermosa landrace maintained in a progeny trial for genetic improvement. Both the multi-locus outcrossing rates (0.95 to 0.99) and the progeny level multi-locus outcrossing rates (0.9 to 1.0) were high, indicating that peach palm is predominantly allogamous. The outcrossing rates among relatives were significantly different from zero (0.101 to 0.202), providing evidence for considerable biparental inbreeding within populations, probably due to farmers planting seeds of a small number of open-pollinated progenies in the same plot. The correlations of paternity estimates were low (0.051 to 0.112), suggesting a large number of pollen sources (9 to 20) participating in pollination of individual fruit bunches. Effective population size estimates suggest that current germplasm collections are insufficient for long-term ex situ conservation. As with most underutilized crops, on farm conservation is the most important component of an integrated conservation strategy.

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.

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

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.

As populações híbridas de pupunha (Bactris gasipaes Kunth) acumularam variabilidade genética provenientes de raças primitivas ao seu redor, o que deveria aumentar sua variabilidade. Para testar esta hipótese, avaliou-se a variabilidade genética de populações híbridas por meio de marcadores RAPD utilizando 176 plantas mantidas no Banco Ativo de Germoplasma do INPA, Manaus-AM, sendo quatro populações híbridas [Belém (n=26); Manaus (n=38); Iquitos, Peru (n=41); Yurimáguas, Peru (n=41)], duas populações silvestres (B. gasipaes variedade chichagui) tipos 1 (n=21) e 3 (n=7), e duas amostras de espécie afim, B. riparia, e compararam-se os parâmetros genéticos com estudos das raças primitivas. Oito iniciadores RAPD geraram 88 marcadores polimórficos e 11 monomórficos. O teste de replicabilidade apresentou uma similaridade de Dice 0,67, considerado aceitável. A heterozigosidade média das populações híbridas foi 0,34 e o polimorfismo foi 87,9%, maiores que nas silvestres (0,31; 74,7%). O dendrograma das similaridades de Dice não apresentou grupos que representassem claramente as populações híbridas. O fluxo gênico entre Iquitos e Yurimáguas (Nm=12,75) e entre Iquitos e Manaus (Nm=9,47) foi alto, enquanto o fluxo entre Belém e Manaus (Nm=7,72) foi menor que o esperado, possivelmente devido à influência da raça Solimões. O alto valor de heterozigosidade em Manaus (0,31) parece ser resultado da união de duas dispersões após a domesticação: a do oeste amazônico, com Iquitos e Yurimáguas, e a do leste amazônico, com Belém, que se juntam em Manaus. No entanto, essas populações não apresentaram acúmulo de variabilidade genética tão expressiva para diferenciá-las das raças primitivas.

The hybrid populations of peach palm (Bactris gasipaes Kunth) have accumulated genetic variability from adjacent landraces, which is expected to increase their variability. To test this hypothesis, 176 plants maintained in the peach palm Germplasm Bank at INPA, including four hybrid populations [Belém (n=26); Manaus (n=38); Iquitos, Peru (n=41); Yurimáguas, Peru (n=41)], two wild populations (B. gasipaes variety chichagui) types 1 (n=21) and 3 (n=7), and two samples of a related species, B. riparia, were genotyped with RAPD markers and compared to adjacent landraces reported in other studies. Eight RAPD primers generated 88 polymorphic and 11 monomorphic markers. The replicability test presented a Dice similarity of 0.67, considered acceptable. The average heterozygosity of the hybrid populations was 0.34 and the polymorphism was 87.9%, greater than in the wild populations (0.31; 74.7%). The dendrogram of Dice similarities did not present groups that clearly correspond to the hybrid populations. The gene flow between Iquitos and Yurimáguas (Nm=12.75), and between Iquitos and Manaus (Nm=9.47) was high, while between Belém and Manaus it was lower than expected (Nm=7.72), probably due to the influence of the Solimões landrace near Manaus. The high value of heterozygosity in Manaus seems to be the result of the union of two dispersals after domestication: the western Amazonian dispersal, with Iquitos and Yurimáguas, and the eastern Amazonian, with Belém, joining in Manaus. However, these hybrid populations did not present an accumulation of genetic variability expressive enough to differentiate them from the landraces.

A pupunheira é uma fruteira nativa da Amazônia com ampla diversidade genética em suas populações silvestres e domesticadas. Este estudo finalizou a caracterização e análise genética dos acessos do germoplasma, usando marcadores moleculares RAPDs. Oito iniciadores geraram 132 marcadores, com 128 polimórficos. A heterozigosidade média foi 0.35 com 95.5% de polimorfismo, similares a outros estudos. A variabilidade dentro (Hs) das raças bem representadas foi 0,32, enquanto a divergência genética (G ST) foi 0,09 entre as quatro raças bem representadas. O fluxo gênico médio foi 5,0, com alto fluxo gênico entre as raças Solimões e Pampa Hermosa (Nm=10,07), e as raças Putumayo e Solimões (Nm=10,73), sendo todas ocidentais. Os dendrogramas das distâncias de Nei e Rogers mostraram que raças bem representadas apresentaram grupos similares às análises anteriores, com as raças Solimões, Putumayo e Pampa Hermosa agrupadas no oeste da Amazônia, e distantes da raça Pará no centro e leste da Amazônia.

Peach palm is a native Amazonian fruit species with broad genetic diversity in its wild and domesticated populations. This study completed the molecular characterization and genetic analysis of the accessions in the Peach Palm genebank, Brazil, using RAPD markers. Eight primers generated 132 markers, of which 128 were polymorphic. Average heterozigosity was 0.35, with 95.5% polymorphism. The genetic variability within the landraces (H S) was 0.32, while genetic divergence (G ST) was 0.09 among the four well-represented landraces. The average gene flow was 5.0, with high gene flow between the Solimões and Pampa Hermosa (Nm=10.07), and between the Putumayo and Solimões (Nm=10.73) landraces, all western landraces. The dendrograms of the Nei's and Rogers' genetic distances of the well-represented landraces presented similar groupings to previous analyses, with the Solimões, Putumayo and Pampa Hermosa landraces grouped together in western Amazonia, and distant from the Pará landrace in central and eastern Amazonia.

In this study, anticoagulant activity was detected in salivary gland homogenates (SGHs) of Thyrsopelma guianense (Diptera: Simuliidae). The SGH yielded 1.07 μg ± 0.03 (n = 15) of total soluble protein per pair of glands. In addition, following SDS-PAGE (12.5% 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.

Zeyheria montana, planta arbustiva da família Bignoniaceae, é uma espécie endêmica do Cerrado e possui atividade anti-câncer, sendo utilizada como estimulante na medicina popular. O objetivo deste estudo foi avaliar a variabilidade genética de oito populações localizadas no estado de São Paulo, utilizando marcadores moleculares de Polimorfismo de DNA Amplificado ao Acaso (RAPD). Após a otimização da reação de amplificação, nove iniciadores selecionados geraram 105 fragmentos RAPD reprodutíveis, sendo que a maioria (60,0%) foi polimórfica. A análise molecular de variância (AMOVA) mostrou que a variabilidade dentro de populações (84,03%) foi maior que entre populações (15,97%). As estimativas de variação fiST (0,1597) indicam estruturação populacional moderadamente alta. O agrupamento por meio de UPGMA, a ordenação pelo NMDS e o teste de Mantel entre as matrizes de distâncias genéticas e geográficas demonstraram que a estruturação encontrada segue um modelo de "ilhas", onde uma única população de tamanho infinito pode ter dado origem às populações atuais de Zeyheria, sem relação com sua posição espacial. Com base nos resultados obtidos foi estruturado um banco de germoplasma de indivíduos, representando a variabilidade da espécie. Adicionalmente, as informações deste estudo são importantes para dar suporte a estratégias de conservação de Z. montana.

Zeyheria montana, an endemic species of the Bignoniaceae family from the Brazilian Cerrado's known for its anti-cancer properties, is widely used as imuno stimulant in the popular medicine and its therapeutic activity must be validated by scientific data. The objective of this work was to evaluate the genetic variability of eight plant populations collected within the state of São Paulo, Brazil, via Random Amplification of Polymorphic DNA (RAPD) used as molecular markers. After an optimized protocol for the amplification reaction, nine selected primers generated 105 reproducible bands, indicating up to 60% polymorphism. Analysis of molecular variance (AMOVA) revealed higher genetic variation within populations (84.03%) than among populations (15.97%). The variation values estimated by phiST (0.160) indicated moderate to high inter population structuration. Levels of similarity inter plants with genetic and geographical distances, estimated by the unweighted pair-group method analysis (UPGMA) clustering and non-metric multidimensional scaling (NMDS) ordination methods and by the Mantel test (-0.2345 p = 0.118) denoted that the structure found follows the island model, which assumes that a single population of infinite size may have initiated the existing populations of Zeyheria montana, with no spatial position correlation. Based on the obtained data, a germplasm bank from individuals representing the species variability was established. Furthermore the information here reported can be of importance to develop strategies for the conservation of Z. montana.

A endonuclease de restrição BliAI, um isoesquisômero de ClaI, que reconhece a seqüência 5'- AT<FONT FACE=Symbol>¯</FONT>CGAT - 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 37ºC e em uma ampla escala de pH e concentração de sais. O peso molecular da enzima purificada corresponde a um valor de kDa 39.

The restriction endonuclease BliAI, an isoschizomer of ClaI, which recognizes the sequence 5'- AT<FONT FACE=Symbol>¯</FONT>CGAT - 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 37ºC 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.

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.

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.

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.

Focal epithelial hyperplasia or Heck's disease is a rare contagious disease caused by human papillomavirus types 13 or 32, initially described among Native American populations. This condition is characterized by the occurrence of multiple small papules or nodules in oral cavity, especially on labial and buccal mucosa and tongue. This report describes the diagnosis of focal epithelial hyperplasia in five Central Amazonian Indians who sought treatment at the Amazonas State Foundation of Tropical Medicine (FMT-AM), using clinical criteria, polymerase chain reaction (PCR) and DNA sequencing.

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. Plant extract preparations, fractionation of promising plant extracts, propagation protocols and definition of in planta studies to maximize product yield of plant species producing active compounds have to be performed to provide a continuing supply of bioactive materials. Chemical characterization of natural compounds, determination of mode of action by kinetics and other spectroscopic methods (MS, X-ray, NMR), as well as in vitro and in vivo biological assays, chemical derivatization, and structure-activity relationships have to be carried out to provide a thorough knowledge on which to base the search for natural compounds or their derivatives with biological activity.