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

Leukocytes respiratory burst and lysozyme level in pacu (Piaractus mesopotamicus Holmberg, 1887)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Crescimento, partição de fotoassimilados e produção de rizomas em taro cultivado sob sombreamento artificial

O cultivo associado é uma alternativa para culturas de ciclo longo e propriedades com limitação de área. Todavia, para o estabelecimento de associações, há necessidade de conhecer a tolerância das espécies ao sombreamento. O trabalho objetivou avaliar o crescimento de parte aérea, partição de fotoassimilados e produção de rizomas em plantas de taro 'Japonês' cultivadas sob intensidades e períodos de sombreamento artificial. Utilizou-se o delineamento de blocos ao acaso, com 13 tratamentos e quatro repetições. Os tratamentos foram constituídos de quatro intensidades de sombreamento (controle = pleno sol; 18; 30 e 50% de sombra, mantidos durante o ciclo todo), além da implementação das intensidades de sombra de 18; 30 e 50%, em três períodos (inicial = 0 a 3; intermediário = 3 a 6 e final = 6 a 9 meses). Aos 60; 90; 120; 150; 180; 210; 240 e 270 dias após o plantio (dat), avaliou-se crescimento de planta e partição de massa entre órgãos e aos 270 dat a produção de rizomas. Plantas sob sombreamento, sobretudo nas maiores intensidades e durante o ciclo todo, apresentaram maior produção de biomassa de parte aérea e de rizomas-mãe e filhos pequenos, em detrimento de rizomas-filho grandes, médios e comerciáveis. A intensidade de 18% de sombra, durante todo o ciclo ou nos períodos inicial e intermediário, foi a que menos afetou o desenvolvimento das plantas e produção de biomassa de rizomas-filho comerciáveis.

Inorganic pyrophosphate-phosphohydrolytic activity associated with rat osseous plate alkaline phosphatase

Purified membrane-bound alkaline phosphatase from rat osseous plate hydrolyzed pyrophosphate in the presence of magnesium ions, with a specific activity of 92.7 U/mg. Optimal apparent pH for pyrophosphatase activity was 8.0 and it remained unchanged on increasing the pyrophosphate concentration. In the absence of magnesium ions the enzyme had a K-m = 88 mu M and V = 36.7 U/mg for pyrophosphate and no inhibition by excess substrate was observed. Pyrophosphatase activity was rapidly destroyed at temperatures above 40 degrees C, but magnesium ions apparently protected the enzyme against danaturation. Sodium metavanadate (Ki = 1.0 mM) was a competitive inhibitor of pyrophosphatase activity, while levamisole (Ki = 8.2 mM) and theophylline (Ki = 7.4 mM) were uncompetitive inhibitors. Magnesium ions (K-0.5 = 1.7 mu M) stimulated pyrophosphatase activity, while cobalt (Ki = 48.5 mu M) and zinc (Ki = 22.0 mu M) ions were non-competitive inhibitors. Manganese and calcium ions had no effect on pyrophosphatase activity. The M-w of the pyrophosphatase: protein was 130 kDa by gel filtration, but a value of 65 kDa was obtained by dissociative gel electrophoresis, suggesting that it was a dimer of apparently identical subunits. These results suggested that pyrophosphatase activity stems from the membrane-bound osseous plate alkaline phosphatase and not from a different protein.

Avaliação da atividade dos inibidores de tripsina após digestão enzimática em grãos de soja tratados termicamente

Este trabalho avalia a reativação dos inibidores de tripsina, após proteólise in vitro, de grãos de soja tratados termicamente. Para a inativação térmica dos inibidores, os grãos foram embebidos em água destilada (1:5 p/v) durante 12 horas e aquecidos em placas sob refluxo por 30 minutos. A reativação dos inibidores foi avaliada em comparação com a atividade das amostras cruas e aquecidas. A digestibilidade in vitro das proteínas variou de 47% ('OC-13') a 59% (Paraná), apresentando uma melhora, em média, de 32,6% com o aquecimento. A atividade dos inibidores de tripsina para os grãos crus variou de 122 a 206 UTI/mg de amostra, e os valores correlacionaram-se negativamente com a porcentagem de digestibilidade (r = -0,9177). Os inibidores tiveram suas atividades totalmente inativadas com o aquecimento dos grãos, os quais apresentaram porcentagem de recuperação, em média, de 40%. Com o aquecimento, a inativação dos inibidores provavelmente ocorre por complexação com os componentes do grão, o que leva à recuperação da atividade com o processo de digestão enzimática.

Projeto, construção e levantamento de desempenho de um concentrador solar cilíndro parabólico com mecanismo automático de rastreamento solar

An cylinder-parabolic solar concentrator is presented to produce steam for different applications. This prototype was built in glass fiber with dimensions that follow a study of optimization of parameters inherent in the optical reflection of sunlight by the surface of reflection and absorption of the same by tubing that leads the fluid of work. The surface of the concentrator of 2.24 m² has been covered by layers of mirror with 1.0 m of lenght and 2.0 cm wide. The absorb tubing consists of a copper tube diameter equal to 28 mm. The concentrator is moving to follow the apparent motion of the sun. It will be presented the processes of manufacturing and assembly of the concentrator proposed, which has as main characteristics the facilities construction and assembly, in addition to reduced cost. Will be presented data from tests performed to produce steam setting up some parameters that diagnose the efficiency of the concentrator. It will be demonstrated the viabilities thermal, economic and of materials of the proposed system.The maximum temperature achieved in the vacuum tube absorber was 232.1°C and average temperature for 1 hour interval was 171.5°C, obtained in a test with automation. The maximum temperature achieved in the output of water was 197.7°C for a temperature of 200.0°C in the absorber tube. The best average result of the water exit temperature to interval of 1 hour was 170.2°C for a temperature of 171.2°C, in the absorber tube, obtained in test with automation. Water exit mean temperatures were always above of the water steaming temperature. The concentrator present a useful efficiency of 38% and a production cost of approximately R$ 450,00 ( $ 160.34)

Estudo térmico de um sistema solar de aquecimento de água residencial para duas configurações de superfície absorvedora

It presents a solar collector to be used in a system for heating water for bathing, whose main characteristics are its low cost and easy manufacturing and assembly. The absorbing surface of the collector is formed by an aluminum plate with eight flaps where they lodge PVC pipes. The catchment area of solar radiation corresponds to 1.3 meters. The collector box was made of wood, is covered by transparent glass and thermal insulation of tire chips and expanded polystyrene (EPS). Absorber tubes were connected in parallel through the use of PVC fittings and fixed to the plate by the use of metal poles and rivets. The entire absorber received paint flat black for better absorption of sunlight. The system worked on a thermosiphon assembly and absorber of the collector has been tested in two configurations: with the tubes facing up, directly exposed to the impact of sunlight and facing down, exchanging heat with the plate by conduction. It was determined the most efficient configuration for the correct purpose. The solar collector was connected to a thermal reservoir, also alternative, low-cost forming the system of solar water heating. We evaluated thermal parameters that proved the viability of the heating system studied

Análise de desempenho de um fogão solar construído a partir de sucatas de antena de tv

Given the growing environmental crisis caused by degradation, mainly due to the use of polluting energy sources, increasing the growing use of renewable energies worldwide, with emphasis on solar energy, an abundant supply and available to everyone, which can be harnessed in several ways: electricity generation; dehydration of food; heating, disinfection and distillation and cooking. The latter has as its primary feature the viability of clean, renewable energy for society, combating ecological damage caused by large-scale use of firewood for cooking foods, use in tropical countries with high solar radiation, and has funding NGOs throughout the world with the goal of achieving low-income population. The proposed project consists of a solar cooker for concentration, working from the reflection of sunlight by a hub that they converge to a focal point at the bottom of the pot, getting lots of heat. The solar cooker under study consists of two elliptical reflecting parabolas made from the recycling of scrap TV antenna, having 0.29 m² of surface area for each antenna, which were covered by multiple mirrors of 2 mm thick and mounted on a metal structure, with correction for the mobility of the apparent movement of the sun. This structure was built with the recycling of scrap metal, possessing a relatively low cost compared with other solar cookers, around US$ 50.00. This cost becomes negligible, since that will involve a great benefit to not have fuel costs for each meal, unlike the use of gas or firewood for cooking food. The tests show that the cooker has reached the maximum temperature of 740 ° C, for boiling water in an average time of 28 minutes, cooking various types of foods such as potatoes, rice and pasta in an average time of 45 minutes and still going as a solar oven, making pizza baking and meat. These cooking times do not differ much from the cooking times on a gas stove, it becomes the solar cooker as a good consumer acceptance, and furthermore not to deliver the same gases that can poison the food as with the wood stove. Proves the viability of using the stove to cook or bake in two daily meals for a family, still presenting a position to improve his performance with the addition of new materials, equipment and techniques

Viabilidade de uso de um fogão solar para cocção de alimentos com parábola refletora fabricada em compósito que utiliza fibras de Carnaúba e resina Ortoftálica

Presents a composite formed by orthophthalic resin and fiber loading of carnauba straw. The fibers were first dried in direct sun exposure and subsequently ground into fodder for the reduction in size. Various formulations of the composite were preliminarily tested by choosing the one presenting the best processability in applying the mold. The composite produced is used for the manufacture of a parabolic surface subsequently coated with mirror segments, flexible plastic, for reflecting the solar rays incident on it. The reflective parable represents the main element of the solar cooker that works with the concentration of sunlight and has dimensions of 1.14 m in diameter and area of 1.0 m². Manufacturing processes and assembly of solar cooker concentration produced are presented. The results of tests for cooking and baking various foods, including rice, pasta, beans, cake, cassava, shrimp, beef, breaded demonstrating the competitiveness of solar cooker studied with other stoves already manufactured and tested in Brazil are presented and in the world. It was also demonstrated the feasibility of the proposed composite for Prototypes manufacture of solar and other structures that do not require great efforts resistance

Degradação luminosa e retenção foliar dos corantes azul brilhante FDC-1 e amarelo tartrasina FDC-5 utilizados como traçadores em pulverizações

Três estudos foram conduzidos no Núcleo de Pesquisas Avançadas em Matologia (NUPAM) pertencente à UNESP/FCA - campus de Botucatu-SP, com o objetivo de avaliar a estabilidade dos corantes Azul Brilhante FDC-1 e Amarelo Tartrasina FDC-5 quanto a diferentes períodos de exposição à luz solar e contato com folhas de Eichhornia crassipes. No primeiro estudo, soluções de 0,3125, 0,625, 1,25, 2,5, 5, 10 e 20 ppm dos corantes Azul Brilhante FDC-1 e Amarelo Tartrasina FDC-5 foram acondicionadas em tubos de quartzo hermeticamente fechados e submetidos a 0, 0,5, 1, 2, 4, 6 e 10 horas de exposição à luz solar e ao escuro. Ao final de cada período, amostras de 10 mL foram retiradas dos tubos e analisadas. No segundo estudo, os tratamentos foram dispostos no esquema fatorial 2x7: duas condições luminosas (escuro e pleno sol) e sete períodos de exposição (0, 0,5, 1, 2, 4, 6 e 10 horas), com seis repetições. Com o auxílio de micropipeta, oito gotas de 5 µL das soluções Azul Brilhante e Amarelo Tartrasina a 4.000 ppm foram depositadas em placas de Petri de vidro. Após o término dos períodos de exposição, as placas foram lavadas com 50 mL de água destilada, com o objetivo de extrair o corante depositado sobre elas. No terceiro estudo, adotaram-se os mesmos tratamentos do segundo experimento, com quatro repetições, porém as soluções foram depositadas sobre as folhas de plantas de Eichhornia crassipes. Foram adotados também os mesmos procedimentos de extração dos corantes após o término dos períodos de exposição. As soluções finais obtidas nos três estudos foram submetidas à leitura óptica de absorbância em espectrofotômetro UV-visível nos comprimentos de onda de 630 e 427 nm, para os corantes Azul Brilhante FDC-1 e Amarelo Tartrasina FDC-5, respectivamente. As várias concentrações das soluções de ambos os corantes não sofreram degradação pela luz solar quando submetidas aos vários períodos de incidência luminosa nos tubos de quartzo (ambiente fechado), visto que as curvas de recuperação apresentaram equações semelhantes àquelas concentrações que foram mantidas no escuro. A mesma estabilidade também foi observada quando os corantes foram submetidos à luz solar em ambiente aberto, ou seja, nas placas de Petri. O corante Amarelo Tartrasina também se apresentou muito estável quando depositado sobre as folhas de E. crassipes, independentemente da exposição ou não à luz solar. Para o corante Azul Brilhante, ocorreram significativas perdas de 7,8 e 18,6% quando esteve depositado na superfície da folha de aguapé pelo período de 10 horas sob condições de escuro e plena luz solar, respectivamente.

Effects of irradiance and spectral composition on the establishment of macroalgae in streams in southern Brazil

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

Polyphenol oxidase from yacon roots (Smallanthus sonchifolius)

Polyphenol oxidase (E.C. 1.14.18.1) (PPO) extracted from yacon roots (Smallanthus sonchifolius) was partially purified by ammonium sulfate fractionation and separation on Sephadex G-100. The enzyme had a molecular weight of 45 490 +/- 3500 da and K-m values of 0.23, 1.14, 1.34, and 5.0 mM for the substrates caffeic acid, chlorogenic acid, 4-methylcatechol, and catechol, respectively. When assayed with resorcinol, DL-DOPA, pyrogallol, protocatechuic, p-coumaric, ferulic, and cinnamic acids, catechin, and quercetin, the PPO showed no activity. The optimum pH varied from 5.0 to 6.6, depending on substrate. PPO activity was inhibited by various phenolic and nonphenolic compounds. p-Coumaric and cinnamic acids showed competitive inhibition, with K-i values of 0.017 and 0.011 mM, respectively, using chlorogenic acid as substrate. Heat inactivation from 60 to 90 degrees C showed the enzyme to be relatively stable at 60-70 degrees C, with progressive inactivation when incubated at 80 and 90 degrees C. The E-a (apparent activation energy) for inactivation was 93.69 kJ mol(-1). Sucrose, maltose, glucose, fructose, and trehalose at high concentrations appeared to protect yacon PPO against thermal inactivation at 75 and 80 degrees C.

POLYPHENOLOXIDASE FROM ATEMOYA FRUIT (ANNONA CHERIMOLA MILL. ANNONA SQUAMOSA L.)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Investigation of the Photodynamic Effects of Curcumin Against Candida albicans

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

Functional significance of eIF5A and its hypusine modification in eukaryotes

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