Atividade anticoagulante, antioxidante e antitumoral de heterofucanas da alga Dictyopteris delicatula (Lamouroux, 1809)

In the present study, six families of sulfated polysaccharides were obtained from seaweed Dictyopteris delicatula (Lamouroux, 1809) and their anticoagulant, antioxidant and antitumor activities were evaluated. All fractions showed anticoagulant activity on aPTT assay, but not on PT assay. Fractions also exhibited total antioxidant activity, superoxide radical scavenging capacity and ferric chelating property. Thus, six fractions (F0.5v, F0.7v, F1.0v, F1.3v, F1.5v e F2.0v) we obtained by proteolytic digestion, followed by acetone fractionation and molecular sieving on Sephadex G-100. Chemical analyses demonstrated that all polysaccharides contain heterofucans composed mainly of fucose, xylose, glucose, galactose, uronic acid, and sulfate. Any fractions changed the PT. However, all fractions were able on double the aPPT on a dose-dependent manner. The heterofucans F0.7v and F1.0v showed low anticoagulant activity while F1.5v presented the most prominent anticoagulant activity .When compared to Clexane®, a low molecular weight heparin, at same concentration F1.5v presented similar anticoagulant activity. The fucans F0.5v and F0.7v at 1.0 mg/mL showed high ferric chelating activity (~45%), whereas fucans F1.3v (0.5 mg/mL) showed considerable reducing power, about 53.2% of the activity of vitamin C. The fucan F1.5v presented the most prominent anticoagulant activity. The best antiproliferative activity was found with fucans F1.3v and F0.7v. However, F1.3v activity was much higher than F0.7v inhibiting almost 100% of HeLa cell proliferation. These fucans have been selected for further studies on structural characterization as well as in vivo experiments, which are already in progress

Caracterização molecular e laboratorial da talassemia beta e da interação hemoglobina s/talassemia beta

Beta thalassemia arises as a consequence of the reduction (β+, β++, βsilent) or absence (β0) of beta globin chain synthesis and results from a number of mechanisms that lead to genetic defects. The inheritance of beta thalassemia is characterized by the existence of heterozygous individuals, compound heterozygotes, homozygotes and those with coinheritance of beta thalassemia allele and other thalassemias and/or hemoglobin variants. The aim of this study was to perform molecular and laboratory characterization of beta thalassemia in heterozygous and homozygous individuals and in those with coinheritance of S beta thalassemia. A total of 48 individuals were included (35 heterozygotes, 4 homozygotes and 9 S beta thalessemia carriers) referred to the Integrated Laboratory of Clinical Analyses of the Federal University of Rio Grande do Norte (UFRN) and the Hematology Ambulatory Facility of the Dalton Barbosa Cunha Hemocenter (Hemonorte Natal, Brazil). Peripheral blood samples form each patient underwent the following laboratory examinations: erythrogram, hemoglobin electrophoresis at alkaline pH, measurements of Hb A2, Fetal Hb and serum ferritin. DNA was extracted using the illustra blood genomicPrep Mini Spin Kit and molecular characterization was performed by the PCR/RFLP technique, which involves digestion with specific restriction enzymes for IVS-1 nt 1 (G®A), IVS-1 nt 6 (T®C) and codon 39 (CAG®TAG) mutations. Of the 35 heterozygotes, 37.1% showed IVS-1 nt 6 mutation, 42.9% IVS-1 nt 1 and 20% were carriers of other mutations not identified by the technique used. The four homozygous patients presented with the IVS-1 nt 6 mutation, while 66.7% of the individuals with S beta thalassemia had the IVS-1 nt 1 mutation. Codon 39 was not detected in any of the patients investigated. Of the thallasemic alleles found, 40.4% were IVS- 1 nt 1, 40.4% IVS-1 nt 6 and 19.2% were not identified. Laboratory data showed that the heterozygotes exhibited microcytosis and hypochromia, evidenced by MCV ranging from 57 to 75fL and MCH from 15.9 to 23.6 pg. Hemoglobin A2 varied between 3.7 and 7.2%. The homogygotes also showed reduced MCV and MCH and elevated HbA2.. Comparison of laboratory data between heterozygous individuals with IVS-1 nt 1 and IVS-1 nt 6 mutations showed that heterozygotes for the IVS1-1 mutation had significantly lower mean MCV and MCH (p = 0.023 and 0.007, respectively) and significantly higher hemoglobin A2 (p < 0.001) when compared to heterozygotes for the IVS-1 nt 6 mutation. PCR/RFLP was useful in identifying the presence or absence of IVS-1 nt 6, IVS-1 nt 1 and codon 39 mutations in most of the patients investigated here. This is the first study conducted in the state of Rio Grande do Norte, Brazil aimed at identifying beta thalassemia mutations and represents an important contribution to the knowledge regarding the molecular profile of beta thalassemia in our country

Digestibilidade da Matéria Seca, Proteína Bruta e Energia Bruta da Silagem de Milho associada ao Concentrado, por meio de ensaios in vivo e in vitro

A study was made to compare dry matter, crude protein and gross energy digestibility of corn silage, associated to concentrate, through in vitro and in vivo digestion techniques. A completely randomized design was used, with two treatments and nine replications, involving three assays. The in vivo digestion assay was conducted with 12 Holstein heifers, on a 24 h permanent sample collection. The in vitro digestion assay was achieved with three collection methods: manual (M), vacuum pump (VP) and nasoesophagic (N). The collection methods VP and M can he used to determine the dry matter digestibility, instead of in vivo method, for the present study ration. The digestion methods influenced the DM, CP and GE digestibility for the studied ration.

Chromic oxide as a digestibility marker for dogs - A comparison of methods of analysis

The study reported compared coefficients of apparent digestibility (CAD) obtained by total collection (TC) and by chromic oxide (Cr2O3) determined by visible absorption spectrometry (VIS) and flame atomic absorption spectrophotometry (FAAS). These quantification methods were also investigated to assess their precision. Nineteen adult dogs housed in individual digestibility cages were fed three commercial diets, each test including five or seven animals. The experiment was carried out in a completely randomized design with three among-subjects factors (foods) and three within-subject factors (methods), and the dogs were the experimental unit. Estimated CAD for all dietary components in the three diets by either chromium quantification methods did not differ and showed fair agreement with TC results (P > 0.05). Coefficients of chromic oxide recoveries in feces were 1.06 +/- 0.044 and 1.01 +/- 0.045 for VIS and FAAS, respectively, values significantly different (P < 0.05), indicating better recovery for FAAS. Significant differences (P<0.05) were found in the mean concentration of Cr2O3 for diets and feces (17 +/- 0.6 and 19 +/- 0.8g/kg for VIS and FAAS, respectively). Coefficient of variation for FAAS was 1.69% and 1.77% for VIS, which did not differ significantly (P > 0.05). Pearson's correlation coefficient (r=0.99; P < 0.01) indicated positive and significant association among the results for VIS and FASS, indicating reasonable precision and agreement of chromic oxide contents determined by these methods. Therefore, FAAS and VIS can be used successfully to determine Cr2O3 in dog foods and feces. (c) 2006 Elsevier B.V. All rights reserved.

Digestão total e parcial de alguns componentes de dietas contendo diferentes níveis de casca de soja e Fontes de nitrogênio, em bovinos

Este experimento foi realizado para avaliar o efeito de dois níveis de casca de soja (CS) e de três fontes de nitrogênio sobre a digestão total e parcial aparentes no rúmen e nos intestinos. Seis novilhos Holandês x Zebu com idade média de 12 meses e 229 kg de PV foram fistulados no rúmen e no abomaso e distribuídos em quadrado latino 6x6 e alimentados individualmente ad libitum. O consumo e as digestibilidades totais e parciais dos nutrientes foram avaliados. Não houve efeito da fonte de proteína sobre o consumo, quantidades de matéria seca (MS), matéria orgânica (MO), proteína bruta (PB), fibra em detergente neutro (FDN) e da fibra em detergente ácido (FDA) no rúmen, no abomaso e nas fezes e sobre os coeficientes de digestibilidades destes componentes. O aumento, porém, do nível de casca de soja elevou os coeficientes de digestibilidade de todos os componentes estudados nos locais do trato digestivo, com exceção nos intestinos, onde o nível de casca de soja não afetou a digestão da PB, FDN e da FDA. Os resultados indicam que a casca de soja pode substituir tanto o milho como, parcialmente, a fração volumosa nas rações de ruminantes. A farinha de subproduto avícola e a amiréia podem substituir o farelo de soja nas rações de bovinos sem reflexos negativos sobre a digestão de seus componentes nutritivos.

Effects of six carbohydrate sources on dog diet digestibility and post-prandial glucose and insulin response

The effects of six extruded diets with different starch sources (cassava flour, brewer's rice, corn, sorghum, peas or lentils) on dog total tract apparent digestibility and glycemic and insulinemic response were investigated. The experiment was carried out on thirty-six dogs with six dogs per diet in a completely randomized design. The diets containing brewer's rice and cassava flour presented the greatest digestibility of dry matter, organic matter and gross energy (p < 0.05), followed by corn and sorghum; pea and lentil diets had the lowest. Starch digestibility was greater than 98% in all diets and was greater for brewer's rice and cassava flour than for lentils and peas diets (p < 0.05). Dogs' immediate post-prandial glucose and insulin responses (AUC <= 30 min) were greater for brewer's rice, corn, and cassava flour diets (p < 0.05), and later meal responses (AUC >= 30 min) were greater for sorghum, lentil and pea diets (p < 0.05). Variations in diet digestibility and post-prandial response can be explained by differences in chemical composition of each starch source including fibre content and starch granule structure. The nutritional particularities of each starch ingredient can be explored through diet formulations designed to modulate glycemic response. However, more studies are required to support these.

Effects of six carbohydrate sources on diet digestibility and postprandial glucose and insulin responses in cats

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

Biodigestão anaeróbia de dejetos de caprinos obtidos nas diferentes estações do ano

Foram utilizados dejetos produzidos por caprinos, em diferentes estádios fisiológicos e submetidos ao mesmo regime alimentar, nas quatro estações do ano. O objetivo foi avaliar o efeito das estações do ano sobre a digestão anaeróbia de resíduos de caprinos em biodigestores modelo batelada com volume útil de 60 L de substrato em fermentação e mantidos sob temperatura ambiente. Foram avaliadas as produções de biogás, as reduções de sólidos voláteis (SV), os potenciais de produção (m³ de biogás/kg de substrato, de estrume, de sólidos totais (ST) ou sólidos voláteis), os números mais prováveis de coliformes totais e fecais, e a composição do biogás. As reduções de SV foram de 38; 34; 33 e 39% para o verão, outono, inverno e primavera, respectivamente. Os totais de biogás produzidos foram de 1,06 m³ no verão, 0,88 m³ no outono, 0,88 m³ no inverno e 0,99 m³ na primavera, e os potenciais de produção médios foram de 0,02 m³ de biogás/kg de substrato e 0,2 m³ de biogás/kg de estrume para todas as estações. As reduções médias de coliformes totais e fecais foram de 99,99% em todas as estações, e os teores máximos de CH4 no biogás foram 88,3; 84,6; 80,6 e 79,2%, para o verão, outono, inverno e primavera, respectivamente.

Desempenho de um aquecedor de água a biogás

Estudos foram desenvolvidos para dimensionar e adaptar o injetor do queimador principal de um aquecedor de água tipo acumulação de 75 L. O diâmetro do injetor foi redimensionado em função da pressão de serviço de 100 mm H2O e poder calorífico inferior do biogás de 21.600 kJ m-3 n, garantindo a manutenção da potência calorífica do equipamento de 20.900 kJ h-1. Os resultados demonstraram que o queimador adaptado operou com biogás adequadamente, com chama estável. A eficiência média do aquecedor foi de 68%, para ganho térmico de 36,7 ºC, correspondendo à temperatura final da água igual a 62,7 ºC, sendo consumido 0,796 m³n de biogás, aquecendo 75 L de água em 72 minutos.

Quali/quantitative characterization of biogas produced in batch digesters supplied with six distinct substrates

Com a finalidade de se analisar quali/quantitativamente o biogás produzido por 6 tipos de substratos, efetuou-se a presente pesquisa no DER/FCAV/UNESP- SP, Brasil. Os substratos utilizados na digestão anaeróbia foram caracterizados como: 1- Esterco de aves de postura (EAP); 2- Esterco de aves de postura triturado (EAPT); 3- Esterco de aves de corte com cama de maravalha (EACM); 4- Esterco de aves de corte com cama de maravalha triturado (EACMT); 5- Esterco de aves de corte com cama de casca de amendoim (EACA), e 6- Esterco de aves de corte com cama de casca de amendoim triturado (EACAT). Dos resultados obtidos, concluiu-se que, em relação à produção acumulada de biogás, o substrato EAPT foi superior aos demais, enquanto EACM apresentou produção acumulada inferior. em geral, os substratos triturados apresentaram maior quantidade de biogás acumulado em relação aos não triturados, exceto o EACAT, com 20,9 m³ inferior ao EACA. O período de produção máxima de biogás teve início entre 45 e 60 dias, começando a decair a partir dos 120 dias. Aos 57 dias após o enchimento dos biodigestores, o gás produzido possuía teores de CH4 superiores a 53%, e a partir dos 99 dias, todos produziram biogás com teores de CH4 superiores a 70%. Os substratos EAP e EAPT apresentaram maiores concentrações de metano no biogás.

Remoção de matéria orgânica, de nutrientes e de coliformes no processo anaeróbio em dois estágios (reator compartimentado seguido de reator UASB) para o tratamento de águas residuárias de suinocultura

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

Parâmetros de dimensionamento para biodigestores batelada operados com dejetos de vacas leiteiras com e sem uso de inóculo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biofertilizer for control of Guignardia citricarpa, the causal agent of citrus black spot

Biofertilizers generated from the anaerobic and aerobic digestion of cattle manure, known as Bio1 and Bio2, respectively, were studied with regard to their microbial composition and effect on the mycelial growth of Phyllosticta citricarpa, the causal agent of citrus black spot (CBS). Two field experiments were conducted to determine the biofertilizer's potential (Bio1) in controlling CBS (2001/2002 and 2002/2003 crops). It was observed that the greatest number of microorganisms was found in the aerobically produced biofertilizer. Bio2 did not inhibit the mycelial growth of P. citricarpa. Mycelial growth of P. citricarpa was inversely proportional to the Bio1 biofertilizer concentration. In the 2001/2002 cropping season, the Biol effect in controlling CBS was directly proportional to its concentration, at the rate of 0 (healthy fruit), with R-2 = 0.88. Biol had a significant effect in controlling CBS, at a concentration of 10%, during the 2001/2002 cropping season, with DI values of 0.246 and 0.229 for the. 10 and 20% doses, respectively, compared to DI of 0.329 for the control. A directly proportional effect of the biofertilizer concentration on the percentage of fruits with a rating of zero was. also observed in the 2002/2003 cropping season, with R-2 = 0.48. However, even at doses higher than in the preceding cropping season, the biofertilizer was less effective, possibly due to a higher occurrence of the disease. Copper oxychloride and combined applications of copper oxychloride and carbendazim plus mancozeb controlled the disease. The possibility of using the biofertilizer as a protective biofungicide to replace copper oxychloride, especially in organic agriculture, should be explored. (c) 2005 Elsevier Ltd. All rights reserved.

Redução de Bactérias Indicadoras de Poluição Fecal em Estrume de Aves de Postura Tratados por Biodigestão Anaeróbia

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

Culture of osteogenic cells from human alveolar bone: A useful source of alkaline phosphatase

The aim of this study was to obtain membrane-bound alkaline phosphatase from osteoblastic-like cells of human alveolar bone. Cells were obtained by enzymatic digestion and maintained in primary culture in osteogenic medium until subconfluence. First passage cells were cultured in the same medium and at 7, 14, and 21 days, total protein content, collagen content, and alkaline phosphatase activity were evaluated. Bone-like nodule formation was evaluated at 21 days. Cells in primary culture at day 14 were washed with Tris-HCl buffer, and used to extract the membrane-bound alkaline phosphatase. Cells expressed osteoblastic phenotype. The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10.0. This enzyme also hydrolyzes ATP, ADP, fructose-1-phosphate, fructose-6-phosphate, pyrophosphate and beta-glycerophosphate. PNPPase activity was reduced by typical inhibitors of alkaline phosphatase. SDS-PAGE of membrane fraction showed a single band with activity of similar to 120 kDa that could be solubilized by phospholipase C or Polidocanol. (c) 2007 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.