Extração de DNA a partir de sangue humano coagulado para aplicação nas técnicas de genotipagem de antígenos leucocitários humanos e de receptores semelhantes à imunoglobulina

O objetivo deste estudo foi padronizar uma metodologia de extração de DNA de alta qualidade a partir de amostras de sangue coagulado. Quarenta e oito amostras de sangue humano coagulado foram utilizadas para a extração de DNA pelo kit comercial EZ-DNA® (Biological Industries, Beit Haemek, Israel), pelo kit de coluna Neoscience® (One Lambda Inc., San Diego, CA) e pelo método modificado de salting out. Apenas o método de salting out foi capaz de extrair altas concentrações de DNA (média, 180ng/µL), as quais foram medidas pelo detector de fluorescência Qubit® (Invitrogen, USA). Este método permitiu a amplificação dos genes HLA (human leukocyte antigens) pela tecnologia PCR-SSO (polymerase chain reaction - specific sequence of oligonucleotides) Luminex, a qual exige DNA de boa qualidade, e de genes KIR (killer cell immunoglobulin-like receptors) pela técnica made in house PCR-SSP (polymerase chain reaction-sequence specific of primers), a qual demanda uma concentração específica de DNA (10ng/µL). Concluímos que a técnica de salting out modificada foi muito eficiente, simples e rápida para a extração de DNA de amostras de sangue humano coagulado, com o objetivo de realizar a genotipagem de genes HLA e KIR.

Immature Aedes mosquitoes colonize Culex quinquefasciatus breeding sites in neighborhoods in the municipality of Olinda, State of Pernambuco

Introduction The present study shows the colonization of Aedes mosquitoes in breeding sites specific for Culex quinquefasciatus in neighborhoods in the municipality of Olinda. Methods Samples were collected between May 2011 and June 2012 from breeding sites positive for Cx. quinquefasciatus by using a ladle and manual suction pump. Results Aedes aegypti (0.12%), Aedes albopictus (0.03%), and Cx. quinquefasciatus (99.8%) were found across the breeding sites. Conclusions The presence of Aedes ssp. in several Cx. quinquefasciatus breeding sites with a heavy load of organic material demonstrates the need to review the concepts and methods used for treatment, as the use of specific larvicide for breeding sites of Culex.

Analysis of human leukocyte antigens class II-DR in Brazilian children and adolescents with systemic lupus erythematosus

OBJECTIVE: To analyze the frequency of human leukocyte antigens class II-DR in children and adolescents with systemic lupus erythematosus. PATIENTS AND METHODS: Fifty-fiveBrazilian systemic lupus erythematosus children and adolescents and 308 healthy individuals were studied. Gender, race, and age of onset of systemic lupus erythematosus were recorded. The human leukocyte antigens typing of class II-DR was carried out by polymerase chain reaction amplification with sequence-specific primers (PCR-SSP). Data were analyzed statistically using the chi square test with Yates' correction, Fisher's exact test, and Bonferroni's correction. RESULTS: Human leukocyte antigen-DR 15 was the most frequently detected antigen in this group of children and adolescents, and it also occurred more frequently in the female group, in children with onset of systemic lupus erythematosus between 0 and 9 years and between 10 to 14 years, and in the Black race group, but these associations were not statistically significants. CONCLUSION: In this group of children and adolescents with a high degree of racial admixture, we could not verify a significant association between human leukocyte antigens class II-DR and systemic lupus erythematosus.

New taxa of south american ficus(moraceae)

Ten new taxa are described: Ficus aripuanensis C.C. Berg & F. Kooy, F. blephariohylla Vásquez Avila, F. cremersiiC.C. Berg, F. insipida Willd. ssp. scabra C.C. Berg, F. jacobii Vázquez Avila, F. laurentana Vázquez Avila, F. leiophylla C.C. Berg, F. piresiana Vázquz Avila & Berg, F. roraimensis C.C. Berg, F. vittataVázquez Avila.

The Simaba guianensis complex in northers South America

With additional data from specimens not studied by previous workers, the confusing Simaba guianensis compex was re-analized. Simaba polyphylla is recognized as a distinct species and two subspecies of S. guianensis are maintained. Simaba polyphylla, S. guianensis ssp. guianensis, and S. guianensis ssp. ecaudata are keyed, described, and illustrated and specimens examined are cited for each. Specimens of S. guianensis ssp. ecaudata show great morphological diversity but can be separeted into three groups. Further collecting may provide information that shows these groups to be worthy of separate taxonomic status.

Lepidoptera: Papilionoidea e Hesperioidea coletados na Ilha de Maracá, Alto Alegre, Roraima, parte do Projeto Maracá, com uma lista complementar de Hesperiidae De Roraima

Como parte do Projeto Maracá, foram efetuadas quatro coletas da Lepidopterofauna (Rhopalocera) da Ilha de Maracá, RR.Foram coletados 2724 exemplares de 453 espécies distribuídas nas seguintes família e subfamilias:PapilionidaePapilionidae - 8 espéciesPieridaePierinae - 4 espéciesColidinae - 8 espéciesDismorphinae - 1 espécieNymphalidaeIthomiidae - 9 espéciesDanainae - 2 espéciesSatyrinae - 37 espéciesBrassolinae - 12 espéciesMorphinae - 2 espéciesNymphalinae - 56 espéciesApaturinae - 2 espéciesCharaxinae - 18 espéciesLibytheidae - 1 espécieLycaenidaeRiodininae - 68 espéciesLycaeninae - 47 espéciesHesperiidaePyrrhopyginae - 4 espéciesPyrginae - 85 espéciesHesperiinae - 94espéciesPythonidae maraca sp. n., Vidius nostra roraimae ssp. n. são descritas. Papias ignarus (Bells, 1931) é revalidada e Papias sobrinus Schaus, 1902 um novo sinônimo de Papias phainis Godman, 1900.

Fenologia e produtividade do araçá-boi (Eugenia stipitata, Myrtaceae) na Amazônia Central

O araçá-boi (Eugenia stipitata McVaugh ssp. sororia McVaugh, Myrtaceae) é uma frutífera nativa da Amazônia Ocidental com potencial para a indústria de sucos e flavorizantes. Embora pouco plantada na Amazônia brasileira devido a sua acidez, é frequentemente cultivada na Amazônia peruana. O conhecimento de sua fenologia pode ajudar no planejamento do manejo do plantio e da comercialização dos frutos. A fenologia de dez plantas, crescendo num latossolo amarelo degradado, foi observada durante cinco anos. O araçá-boi geralmente floresceu e frutificou três vezes ao longo do ano e sempre teve pelo menos um pico de floração forte durante a estação seca (julho a setembro) e um pico de frutificação mais acentuado na estação chuvosa (janeiro a março). A floração é um evento complexo e demorado que pode durar de dois a três meses, embora o período entre o aparecimento do botão floral até a antese do flor é curto (~15 dias) e o período entre a antese e a maturação dos frutos dura 50 a 60 dias. As regressões múltiplas usadas para determinar o efeito das variáveis climáticas na floração e frutificação não apresentaram altos coeficientes de determinação, embora os modelos tenham sido significantes, provavelmente porque o araçá-boi floresce várias vezes durante o ano e ainda não se sabe qual o estímulo mais importante para iniciar o processo. O vingamento dos frutos variou de menos de 5% a aproximadamente 15%. O peso médio dos frutos avaliados em janeiro de 1988 foi 135 g, com 77% de polpa. Ao longo do período, estimou-se que as dez plantas produziram cm média 1000 frutos/ano, com uma mediana de 890 frutos/ano. Os insetos visitantes eram principalmente abelhas, especialmente Apis mellifera, Eulaema mocsaryi e Ptilotrigona lurida.

Conforto ambiental de bezerros bubalinos (Bubalus bubalis Linnaeus, 1758) em sistemas silvipastoris na Amazônia Oriental

Foi analisado o efeito de dois sistemas silvipastoris nos índices de conforto ambiental e alterações nos parâmetros fisiológicos de bezerros bubalinos criados na Embrapa Amazônia Oriental, Belém-PA (clima Afi), no Período 1 (abril a setembro/2007) e Período 2 (outubro/2007 a março/2008). Foram inseridos onze bezerros no Sistema Silvipastoril 1 (SSP1), que apresentava sombreamento útil nas pastagens de 18 a 21%, e oito no Sistema Silvipastoril 2 (SSP2), sem sombreamento, com lago para banho. Foram mensuradas as variáveis fisiológicas: temperatura retal (TR), frequência respiratória (FR), temperatura da pele (TP), e calculados o Índice de Temperatura e Umidade (ITU) e Índice de Conforto de Benezra (ICB), nos dois períodos experimentais, comparados pelo Teste Tukey (P < 0,05). O ITU apresentou diferença estatística entre horários (P < 0,05) e período do ano (P < 0,05), e oscilou de 73,5 ± 1,3 até 82,2 ± 0,8. A TR apresentou diferença estatística entre horários e períodos do ano (P < 0,05), com amplitude de 38,3 ± 0,26 a 39,3 ± 0,38 °C. A FR apresentou diferença significativa entre horários (P < 0,05), com amplitude de 32,2 ± 9,2 a 56,5 ± 19,0 mov min-1, consideradas acima dos níveis normais, enquanto a TP foi diferente estatisticamente entre períodos e horários (P < 0,05) e variou de 23,6 ± 8,3 a 31,7 ± 5,4 °C. Nos Períodos 1 e 2 e nos dois SSP's, os ICBs estiveram acima do valor ideal, variando de 2,46 ± 0,33 a 3,31 ± 0,62 (SSP1) e 2,42 ± 0,30 a 3,45 ± 0,66 (SSP2).

Estudo comparativo entre silagem de milho e cana taquara na alimentação suplementar de vacas leiteiras

The A. A. compare corn silage (Zea mays L.) with sugar cane (Saccharum sinensis Roxb.) in the supplementary feeding of dairy cow. Both the feeds were studied in relation to the following points: composition and nutritive value; influence of milk production, milk fat, milk acidity and body weight; cost of production. Both corn silage and sugar cane were analysed by ordinary methods, and their digestibility was determined by means of digestibility coefficients; their composition and nutritive value are, practically, equivalent, but silage showed slight superiority. The feeding experiment was carried out with two groups of six Holstein Friesian cows each, of the Escola Superior de Agricultura "Luiz de Queiroz" herd. Both groups were fed with the same basic concentrates mixture, calculeted according to MORRISON. During the various periods of the experiment, only the roughage supplement varied. The supplementary feeding consisted of 15 kg of chopped sugar cane or corn, silage, per day and per cow, given in two daily meals in the barn. At 4,30 p.m., the cows are set free in the field, where they pass the over night. The experiment was divided into six periods, in which there was a gradative change of the supplementary feeding between the two groups. The milk was weighed every day; the analysis of milk fat and acidity and the weighing of the animals, were made only on the first three days of every week. The analysis of data showed that: a) Milk production was increased significantly by silage feeding; b) The ri was not any influence on milk fat; c) The silage caused higher milk acidity; d) The sugar cane gave a greater increase of body weight. The cost of production of corn silage was 2,12 time higher, than sugar cane, hence, although the silage gave a higher milk production, its use is not economical, compared with sugar cane, in our conditions.

Diversity of harvestmen (Arachnida, Opiliones) in Parque da Onça Parda, southeastern Brazil

The environment most diverse in harvestmen species is the Atlantic Forest of São Paulo. However, there remains a lack of studies regarding their communities in certain regions. Among these regions is one south of the Paranapiacaba mountain range in the state of São Paulo, the Parque da Onça Parda (POP). Through nocturnal collections and pitfall traps, the region's harvestmen community has been studied. The observed richness of this site included 27 species, with dominance of three species: Holcobunus nigripalpis Roewer, 1910, Neosadocus maximus (Giltay, 1928) and Munequita sp., accounting for 68.4% of harvestmen abundance. This makes the diversity of POP more similar to the semideciduous Atlantic Forest communities of the interior than to those of the Coastal Atlantic Forest that contains the park. Its geographic location places it within the Southern São Paulo State (SSP) area of endemism, along with the Parque Turístico do Alto Ribeira (PETAR), with which it shares up to 12% similarity regarding harvestmen fauna. Richness and abundance of harvestmen were positively related to temperature and humidity. The period of animal activity (as measured by abundance and richness) varied throughout the night, being highest in the early hours during both studied seasons (summer and winter).

Cirrose hepática enzoótica no cavalo

As lesões histopatológicas observadas no fígado de 12 cavalos de procedências diversas, descritas neste trabalho, sugerem a interferência de um mesmo agente etiológico, possìvelmente uma intoxicação crônica provocada pela infestão de vegetais do gen. Senecio ou de plantas contendo alcalóides afins. Nota: - Já estava redigido o presente trabalho, quando deparamos com o artigo de VANEK (Vanek, J., 1958), Vergiftung mit Kreuskraut (Senecio) als ursache der Zdárer Pferdseuche. - Schveiz. Z. Allg. Path. Bakt., 21:821-848), onde o autor, baseado em dados clínicos, anatõmicos, botânicos, toxicológicos e experimentais, conclui que a doença dos cavalos de Zdar é devida a uma intoxicação crônica pelos alcalóides do Senecio erraticus ssp. barbaraeifolius. As alterações histopatológicas descritas no referido trabalho, coincidem com as observadas nos nossos casos.

A genitália externa dos machos em quatro espécies do gênero Sirthenea Spinola, 1837 (Hemiptera, Reduviidae, Peiratinae)

Com o intuito de acrescentar parâmetros à taxonomia do gênero Sirthenea Spinola, 1837, realizamos uma análise morfológica comparativa da genitália externa dos machos de quatro espécies: S. carinata (Fabricius, 1798), S. stria (Fabricius, 1794), S. flavipes (Stal, 1855) e S. amazona Stal, 1866. Foram evidenciadas 10 estruturas (8º esternito, processo do pigóforo, parâmeros, falosoma e seus quatro processos e os dois processos do endosoma) que permitem diferenciar as quatro espécies. Uma descrição resumida da morfologia geral de cada espécie é adicionada.

Filariopsis barretoi (travassos, 1921) (Nematoda: metastrongyloidea) lung parasite of primates from South America - taxonomy, synonyms and pathology

Nematodes and fragments of lungs from Cebus ssp., Callithrix jacchus (l.) and Saimiri sciureus (L.) were studied. The worms from Cebus and Callithrix must be called Filariopsis barretoi (Travassos, 1921). The names Filariopsis arator Chandler, 1931 and Filaroides cebi Gebauer, 1933 are synonymized to F. barretoi. The status of Filariopsis gordius (Travassos, 1921) remains uncertain. The pathology is described. The parasites are located in the pulmonary paranchyma, near the pleural surface, constituting nodules.

Diterpenoids from Azorella compacta (Umbelliferae) active on Trypanosoma cruzi

The anti-Trypanosoma cruzi activity of natural products isolated from Azorella compacta was evaluated, with particular emphasis on their effect against intracellular amastigotes. Five diterpenoids from A. compacta derived from mulinane and azorellane were isolated and identified. Only two products, named azorellanol (Y-2) and mulin-11,3-dien-20-oic acid (Y-5), showed trypanocidal activity against all stages of T. cruzi including intracellular amastigotes. At 10 µM, these compounds displayed a strong lytic activity. It ranged from 88.4 ± 0.6 to 99.0 ± 1 % for all strains and stages evaluate, with an IC50 /18 h values of 20-84 µM and 41-87 µM, respectively. The development of intracellular amastigotes was also inhibited by nearly 60% at 25 µM. The trypanocidal molecules Y-2 and Y-5 did show different degrees of cytotoxicity depending on the cell line tested, with an IC50 /24 h ranging from 33.2 to 161.2 µM. We evaluated the effect of diterpenoids against intracellular T. cruzi forms by immunofluorescent identification of a specific membrane molecular marker (Ssp-4 antigen) of the T. cruzi amastigote forms. The accuracy and reproducibility of the measurements were found to be outstanding when examined by confocal microscopy.

Association of cytokine genetic polymorphism with hepatites B infection evolution in adult patients

The infection by the hepatitis B virus (HBV) has different forms of evolution, ranging from self-limited infection to chronic hepatic disease. The objective of this study was to evaluate the influence of cytokine genetic polymorphisms in the disease evolution. The patients were divided into two groups, one with chronic HBV (n = 30), and the other with self-limited infection (n = 41). The genotyping for TNF (-308), TGFB1 (+869, +915), IL-10 (1082, -819, and -592), IL-6 (-174), and IFNG (+874) was accomplished by the PCR-SSP (polymerase chain reaction with sequence specific primers technique using the One Lambda kit. Although no statistically significant differences were found between the groups, the combination of TNF -308GG and IFNG +874TA was found in a lower frequency in chronic patients than in individuals with self-limited infection (26.7 versus 46.3%; P = 0.079; OR = 0.40; IC95% = 0.14-1.11). In chronic patients with histological alterations it was not observed the genotype TGFB1+869 C/C, against 24.4% in the self limited infection group (100 versus 75.6%; P = 0.096; OR = 7.67; IC95% = 0.42-141.63). Further studies in other populations, and evaluation of a greater number of individuals could contribute for a better understanding of the cytokine genetic polymorphism influence in HBV infection evolution.