Poder calorífico de algumas substâncias alimentares da cozinha paulista

Apresentamos neste trabalho os primeiros resultados de experiências de nutriição, que sob vários aspectos encetamos no Instituto de Higiene

Production and characterization of collagenase by Penicillium sp. UCP 1286 isolated from Caatinga soil

A new Penicillium sp. strain isolated from the soil of Caatinga, a Brazilian Biome (UCP 1286) was selected for collagenase production. Fermentation system allowing obtention of collagenolytic activity about 2.7 times higher than existing data, with the highest values of collagenolytic and specific activity (379.80 U/mL, 1460.77 U/mg, respectively), after 126 hours. Applying a factorial design, enzyme production was increased by about 65% compared to the preliminary results. The factorial design demonstrated the existence of two factors with statistical significance on the production of the enzyme: pH and temperature, both with negative effects. Enzyme was found to be more active at pH 9.0 and 37 °C, and also to be very stable in comparison with the collagenase produced by other microorganisms. The enzyme seems to belong to collagenolytic serine proteases family. Concerning the substrate specificity, it was observed that the highest enzyme activity corresponds to azocoll, there was no relevant activity on azocasein and the enzyme showed to be more specific to type V collagen and gelatin than the commercial colagenase produced by Clostridium histolyticum. Major band observed at electrophoresis was approximately 37 kDa. Zymogram analysis confirmed the collagenolytic activity. All data indicates this enzyme as promising biotechnology product.This work was supported by Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE) (IBPG-0137-2.08/12) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Sara Silvério also acknowledges her post-doc grant (SFRH/BPD/88584/2012) from FCT (Fundação para a Ciência e a Tecnologia), Portugal

Purification and characterization of a collagenase from Penicillium sp. UCP 1286 by polyethylene glycol-phosphate aqueous two-phase system

Collagenases are proteolytic enzymes capable of degrading both native and denatured collagen, reported to be applied in industrial, medical and biotechnological sectors. Liquid-liquid extraction using aqueous two-phase system (ATPS) is one of the most promising bioseparation techniques, which can substitute difficult solid-liquid separation processes, offering many advantages over conventional methods including low-processing time, low-cost material and low-energy consumption. The collagenase produced by Penicillium sp. UCP 1286 showed a stronger affinity for the bottom salt-rich phase, where the highest levels of collagenolytic activity were observed at the center point runs, using 15.0% (w/w) PEG 3350 g/mol and 12.5% (w/w) phosphate salt at pH 7.0 and concentration. The enzyme was characterized by thermal stability, pH tolerance and effect of inhibitors, showing optimal collagenolytic activity at 37 °C and pH 9.0 and proved to be a serine protease. ATPS showed high efficiency in the collagenase purification, confirmed by a single band in SDS/PAGE, and can in fact be applied as a quick and inexpensive alternative method.This work was supported by the National Council of Technological and Scientific Development (CNPq) and Coordination for the Improvement of Higher Education Personnel (CAPES). Sara Silvério also acknowledges her post-doc grant (SFRH/BPD/88584/2012) from FCT (SFRH/BPD/88584/2012) (Portuguese Science and Technology Foundation), Portugal

Morphological characterization of coriander genotypes

Poucas cultivares de coentro (Coriandrum sativum L.) estão disponíveis aos produtores e, em algumas regiões, cultivam-se genótipos de origem desconhecida, mantidos pelos próprios hortelãos. Para conservar as cultivares tradicionais é necessário não só coletá-las e conservá-las, mas também caracterizá-las. Dessa forma, este trabalho teve por objetivo determinar caracteres morfológicos para diferenciar genótipos de coentro no processo de registro de cultivares. Foram realizados dois experimentos, ambos em blocos casualizados, com cinco repetições. As cultivares Americano, Asteca, Palmeira, Português, Santo, Supéria, Tabocas, Tapacurá e Verdão, além da linhagem HTV-9299 foram analisadas para caracteres de fruto, plântula, folhas, folíolos e flores. Os dados obtidos foram submetidos à análise de variância univariada. A cultivar Palmeira apresentou frutos de maior tamanho e massa. Já para antocianina a cultivar Americano apresentou menor intensidade, ou praticamente ausência do pigmento tanto nas plântulas quanto nas plantas, ao contrário da cultivar Tabocas que apresentou maior intensidade. As cultivares Americano, Tabocas, Palmeira, Asteca, Verdão e Tabocas e a linhagem HTV-9299 possuem maior número de folhas basais, no entanto, as cultivares que produziram maior quantidade de massa por planta foram Tabocas, Palmeira, Verdão, HTV-9299 e Tapacurá. A cultivar Americano possui folíolo verde claro pouco amarelado, a linhagem HTV-9299 folíolo verde e a cultivar Tabocas folíolo verde escuro. Os descritores morfológicos com maior poder discriminatório para o grupo de dez cultivares avaliadas foram tamanho e massa de fruto; antocianina na plântula e planta; número de folhas basais; comprimento da quinta folha; tamanho e coloração de folíolo; número de dias para pendoamento e abertura da primeira flor e massa da planta fresca.Few coriander cultivars (Coriandrum sativum L.) are available to producers and, in some regions, are grown genotypes of unknown origin, kept by the growers themselves. To conserve such traditional crops it is necessary to collect, save, and characterize them. Thus, this study was to determine morphological characters to differentiate coriander genotypes aiming the registration of cultivars. Two trials were carried out, and the experimental design used in both was in completely randomized blocks with five replications. Americano, Asteca, Palmeira, Português, Santo, Superia, Tabocas, Tapacurá, and Verdão cultivars, and the line HTV-9299 were characterized for seedling, leaflets, leaves, fruits, and flowers. The data were subjected to a univariate ANOVA. The fruits of cv. Palmeira exhibited greater size and mass. As regards the presence of anthocyanins in seedlings and plants, cv. Americano showed the lowest intensity or slight pigmentation while cv. Tabocas had the highest pigmentation. The cultivars Americano, Palmeira, Asteca, Verdão, and Tabocas and the line HTV-9299 displayed higher number of basal leaves, however, the genotypes that produced larger amount of mass per plant were Tabocas, Palmeira, Verdão, HTV-9299, and Tapacurá. The green tone of leaflets showed differences among cultivars. So, cv. Americanos leaflets exhibited a light green, slightly yellowish color, HTV-9299 green, and Tabocas dark green. The morphological traits with higher discriminatory power for the group of the ten evaluated cultivars were: size and weight of fruits; presence or absence of anthocyanin in seedlings and plants; number of basal leaves; length of the fifth leaf; size and color of leaflets; number of days to bolting initiation, and opening of the first flower, and plant fresh mass

Genetic variability of half-sib progenies of coriander

Grande número de produtores envolve-se com o cultivo do coentro durante todo o ano, tornando a cultura importante social e economicamente. Praticamente em toda a região Nordeste utiliza-se a cultivar Verdão. Estudos da variabilidade genética do coentro são importantes, tendo em vista o melhor planejamento de programas de melhoramento genético. Desta forma, este trabalho teve como objetivo quantificar a variabilidade genética de características agronômicas do coentro cultivar Verdão, avaliando-se progênies de meios-irmãos potencialmente úteis no melhoramento genético. O trabalho foi desenvolvido na Universidade Federal Rural de Pernambuco, em casa de vegetação, em blocos casualizados, com cinco repetições e parcelas de 28 plantas, colocadas em dois vasos. Os tratamentos foram compostos por 55 progênies de meios-irmãos da cultivar Verdão. O teste t detectou significância a 1 e a 5% de probabilidade entre as correlações genotípicas, fenotípicas e ambientais. A herdabilidade no sentido amplo variou de 7,19 (peso médio de plantas) a 81,09 (número de plantas pendoadas). Para a razão entre os coeficientes de variação genético e ambiental (CVg /CVe), obteve-se de 0,27 (peso médio de plantas) a 2,07 (número de plantas pendoadas), indicando que a seleção contra o pendoamento apresenta as condições mais favoráveis em termos de ganhos genéticos imediatos. A correlação genotípica entre altura de plantas não pendoadas e número de plantas pendoadas foi alta e significativa.A large number of producers are involved in the cultivation of coriander throughout the year in Brazil, making it a crop of social and economic importance. Throughout nearly the entire Northeast Region, cultivar Verdão is used. Genetic variability studies on coriander are important for the proper planning of genetic improvement programs. The aim of the present study was to quantify the genetic variability for agronomic characteristics in cv. Verdão to contribute information toward genetic improvement. The study was developed at Universidade Federal Rural de Pernambuco, Pernambuco State, Brazil, in a greenhouse, using randomized blocks of 28 plants in two pots and five replications. The treatments were composed of 55 half-sib progenies from cultivar Verdão. The t test detected significance at 1 and 5% likelihood between the genotypic, phenotypic and environmental correlations. Broad sense heritability ranged from 7.19 for average plant weight to 81.09 for number of bolted plants. The ratio between genetic and environmental coefficients of variation (CVg/CVe) ranged from 0.27 (average weight) to 2.07 (number of bolted plants), indicating that selection against bolting presents more favorable conditions in terms of immediate genetic gains. The genotype correlation between height of non-bolted plants and the number of bolted plants was considered strong (0.86) and highly significant (t test), enabling simultaneous gains from selection

Exposure of Phosphatidylserine on Leishmania amazonensis Isolates Is Associated with Diffuse Cutaneous Leishmaniasis and Parasite Infectivity

Diffuse cutaneous leishmaniasis (DCL) is a rare clinical manifestation of leishmaniasis, characterized by an inefficient parasite-specific cellular response and heavily parasitized macrophages. In Brazil, Leishmania (Leishmania) amazonensis is the main species involved in DCL cases. In the experimental model, recognition of phosphatidylserine (PS) molecules exposed on the surface of amastigotes forms of L. amazonensis inhibits the inflammatory response of infected macrophages as a strategy to evade the host immune surveillance. In this study, we examined whether PS exposure on L. amazonensis isolates from DCL patients operated as a parasite pathogenic factor and as a putative suppression mechanism of immune response during the infection. Peritoneal macrophages from F1 mice (BALB/c×C57BL/6) were infected with different L. amazonensis isolates from patients with localized cutaneous leishmaniasis (LCL) or DCL. DCL isolates showed higher PS exposure than their counterparts from LCL patients. In addition, PS exposure was positively correlated with clinical parameters of the human infection (number of lesions and time of disease) and with characteristics of the experimental infection (macrophage infection and anti-inflammatory cytokine induction). Furthermore, parasites isolated from DCL patients displayed an increased area in parasitophorous vacuoles (PV) when compared to those isolated from LCL patients. Thus, this study shows for the first time that a parasite factor (exposed PS) might be associated with parasite survival/persistence in macrophages and lesion exacerbation during the course of DCL, providing new insights regarding pathogenic mechanism in this rare chronic disease

The Linkages Between Photosynthesis, Productivity, Growth and Biomass in Lowland Amazonian Forests

Understanding the relationship between photosynthesis, net primary productivity and growth in forest ecosystems is key to understanding how these ecosystems will respond to global anthropogenic change, yet the linkages among these components are rarely explored in detail. We provide the first comprehensive description of the productivity, respiration and carbon allocation of contrasting lowland Amazonian forests spanning gradients in seasonal water deficit and soil fertility. Using the largest data set assembled to date, ten sites in three countries all studied with a standardized methodology, we find that (i) gross primary productivity (GPP) has a simple relationship with seasonal water deficit, but that (ii) site-to-site variations in GPP have little power in explaining site-to-site spatial variations in net primary productivity (NPP) or growth because of concomitant changes in carbon use efficiency (CUE), and conversely, the woody growth rate of a tropical forest is a very poor proxy for its productivity. Moreover, (iii) spatial patterns of biomass are much more driven by patterns of residence times (i.e. tree mortality rates) than by spatial variation in productivity or tree growth. Current theory and models of tropical forest carbon cycling under projected scenarios of global atmospheric change can benefit from advancing beyond a focus on GPP. By improving our understanding of poorly understood processes such as CUE, NPP allocation and biomass turnover times, we can provide more complete and mechanistic approaches to linking climate and tropical forest carbon cycling

Cooperation between Apoptotic and Viable Metacyclics Enhances the Pathogenesis of Leishmaniasis

Mimicking mammalian apoptotic cells by exposing phosphatidylserine (PS) is a strategy used by virus and parasitic protozoa to escape host protective inflammatory responses. With Leishmania amazonensis (La), apoptotic mimicry is a prerogative of the intramacrophagic amastigote form of the parasite and is modulated by the host. Now we show that differently from what happens with amastigotes, promastigotes exposing PS are non-viable, non-infective cells, undergoing apoptotic death. As part of the normal metacyclogenic process occurring in axenic cultures and in the gut of sand fly vectors, a sub-population of metacyclic promastigotes exposes PS. Apoptotic death of the purified PS-positive (PSPOS) sub-population was confirmed by TUNEL staining and DNA laddering. Transmission electron microscopy revealed morphological alterations in PSPOS metacyclics such as DNA condensation, cytoplasm degradation and mitochondrion and kinetoplast destruction, both in in vitro cultures and in sand fly guts. TUNELPOS promastigotes were detected only in the anterior midgut to foregut boundary of infected sand flies. Interestingly, caspase inhibitors modulated parasite death and PS exposure, when added to parasite cultures in a specific time window. Efficient in vitro macrophage infections and in vivo lesions only occur when PSPOS and PS-negative (PSNEG) parasites were simultaneously added to the cell culture or inoculated in the mammalian host. The viable PSNEG promastigote was the infective form, as shown by following the fate of fluorescently labeled parasites, while the PSPOS apoptotic sub-population inhibited host macrophage inflammatory response. PS exposure and macrophage inhibition by a subpopulation of promastigotes is a different mechanism than the one previously described with amastigotes, where the entire population exposes PS. Both mechanisms co-exist and play a role in the transmission and development of the disease in case of infection by La. Since both processes confer selective advantages to the infective microorganism they justify the occurrence of apoptotic features in a unicellular pathogen