Monoamine oxidase inhibitory activities of indolylalkaloid toxins from the venom of the colonial spider Parawixia bistriata: Functional characterization of PwTX-I

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

Development of the insulin secretion mechanism in fetal and neonatal rat pancreatic B-cells: response to glucose, K+, theophylline, and carbamylcholine

We studied the development of the insulin secretion mechanism in the pancreas of fetal (19- and 21-day-old), neonatal (3-day-old), and adult (90-day-old) rats in response to stimulation with 8.3 or 16.7 mM glucose, 30 mM K+, 5 mM theophylline (Theo) and 200 µM carbamylcholine (Cch). No effect of glucose or high K+ was observed on the pancreas from 19-day-old fetuses, whereas Theo and Cch significantly increased insulin secretion at this age (82 and 127% above basal levels, respectively). High K+ also failed to alter the insulin secretion in the pancreas from 21-day-old fetuses, whereas 8.3 mM and 16.7 mM glucose significantly stimulated insulin release by 41 and 54% above basal levels, respectively. Similar results were obtained with Theo and Cch. A more marked effect of glucose on insulin secretion was observed in the pancreas of 3-day-old rats, reaching 84 and 179% above basal levels with 8.3 mM and 16.7 mM glucose, respectively. At this age, both Theo and Cch increased insulin secretion to close to two-times basal levels. In islets from adult rats, 8.3 mM and 16.7 mM glucose, Theo, and Cch increased the insulin release by 104, 193, 318 and 396% above basal levels, respectively. These data indicate that pancreatic B-cells from 19-day-old fetuses were already sensitive to stimuli that use either cAMP or IP3 and DAG as second messengers, but insensitive to stimuli such as glucose and high K+ that induce membrane depolarization. The greater effect of glucose on insulin secretion during the neonatal period indicates that this period is crucial for the maturation of the glucose-sensing mechanism in B-cells.

Growth factors and glucose homeostasis in diabetic rats: effects of exercise training

To investigate the alterations of glucose homeostasis and variables of the insulin-like growth factor-I (IGF- 1) growth system in sedentary and trained diabetic (TD) rats, Wistar rats were divided into sedentary control (SC), trained control (TC), sedentary diabetic (SD), and TD groups. Diabetes was induced by Alloxan (35 mg kg(-1) b.w.). Training program consisted of swimming 5 days week(-1), 1 h day(-1), during 8 weeks. Rats were sacrificed and blood was collected for determinations of serum glucose, insulin, growth hormone (GH), IGF-1, and IGF binding protein-3(IGFBP-3). Muscle and liver were removed to evaluate glycogen content. Cerebellum was extracted to determinate IGF-1 content. Diabetes decreased serum GH, IGF-1, IGFBP-3, liver glycogen, and cerebellum IGF-1 peptide content in baseline condition. Physical training recovered liver glycogen and increased serum and cerebellum IGF-1 peptide in diabetic rats. Physical training induces important metabolic and hormonal alterations that are associated with an improvement in glucose homeostasis and serum and cerebellum IGF-1 concentrations. Copyright (C) 2009 John Wiley & Sons, Ltd.

Exercise test and glucose homeostasis in rats treated with alloxan during the neonatal period or fed a high calorie diet

Background: Animal models appear well-suited for studies into the role of exercise in the prevention of non-insulin-dependent diabetes mellitus (NIDDM). The aim of the present study was to analyze glucose homeostasis and blood lactate during an exercise swimming test in rats treated with alloxan during the neonatal period and/or fed a high calorie diet from weaning onwards.Methods: Rats were injected with alloxan (200 mg/kg, i.p.) or vehicle (citrate buffer) at 6 days of age. After weaning, rats were divided into four groups and fed either a balanced diet or a high-caloric diet as follows: C, control group (vehicle + normal diet); A, alloxan-treated rats fed the normal diet; H, vehicle-treated rats fed the high-caloric diet; and HA, alloxan-treated rats fed the high-caloric diet.Results: Fasting serum glucose levels were higher in groups A and AH compared with the control group. The Homeostatic Model Assessment index varied in the groups as follows: H > A > HA = C. There were no differences in free fatty acids or blood lactate concentrations during the swim test.Conclusions: Alloxan-treated rats fed a normal or high-caloric diet have the potential to be used in studies analyzing the role physical exercise plays in the prevention of NIDDM.

Crystallization and preliminary X-ray diffraction analysis of the catalytic subunit of ADP-glucose pyrophosphorylase from potato tuber

ADP-glucose pyrophosphorylase is the key regulatory enzyme in the biosynthesis of starch in plants and glycogen in bacteria. The enzyme from potato tuber is comprised of a regulatory subunit and a catalytic subunit and is present as a heterotetramer (alpha(2)beta(2)) the catalytic subunit from potato tuber (50 kDa) was crystallized in four different forms, two of which are suitable for structural studies. A tetragonal crystal form obtained in the presence of the substrate analog Cr-ATP diffracted to 2.2 Angstrom and belongs to space group P4(1) (or its enantiomorph), with unit-cell parameters a = b = 110.57, c = 190.14 Angstrom. A second crystal form obtained diffracted to 2.8 Angstrom and belongs to space group PZ, with unit-eel parameters a = 80.06, b = 138.84, c = 92.20 Angstrom, beta = 112.40 degrees. As this protein displays no significant homology to any currently known protein structure, a search for heavy-atom derivatives has been initiated.

Antimicrobial activity of an L-amino acid oxidase isolated from Bothrops leucurus snake venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The structure of a native L-amino acid oxidase, the major component of the Vipera ammodytes ammodytes venomic, reveals dynamic active site and quaternary structure stabilization by divalent ions

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

Crystallization and preliminary X-ray diffraction analysis of an l-amino-acid oxidase from Bothrops jararacussu venom

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

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

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

Glucose metabolism in rats submitted to skeletal muscle denervation

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

Efeito comparativo do calor, S0(2) e ácido ascórbico na atividade da polifenol oxidase e peroxidase de algumas frutas e hortaliças

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

TEOR DE VITAMINA C, ATIVIDADE DE ASCORBATO OXIDASE E PERFIL SENSORIAL DE MANGA (Mangífera índica L.) VAR. HADEN, DURANTE O AMADURECIMENTO

The chemical and biochemical composition of mango, varies according to the cultivation conditions, variety and maturation state, generally containing a high level of ascorbic acid. In order to establish the correlation between the activity of the ascorbate oxidase [E.C.1.10.3.3], and ascorbic acid level in the ripening process of the Haden mango (Mangífera índica L.), sample of the fruits related to hard green stage (zero), 2, 4, 6, 8, 10, 12 and 14 days stored at 20 ± 2oC, were tested. The samples were obtained by cutting small cubes of 8 cm3 from pulps of 8 mangoes with texture without significant difference (p£0.05) at Magness-Taylor pressure tester scale. In each sample the activity of ascorbate oxidase was followed, in order to check its participation in possible substrate losses during the ripening fruits. The ascorbic acid level and sensory profile also was determined periodically during the ripening period. The enzymatic activity was spectrophotometrically determined at 245 nm and 30oC. The ascorbic acid was analyzed according modified AOAC methodology, and sensory analysis by descriptive quantitative analysis. Data were analyzed using correlation analysis, analysis of variance (ANOVA), Tukey's test, principal component analysis and stepwise discriminant analysis. During the ripening, the ascorbate oxidase activity increased (from 0 to 5.0 x 10-1 U/ml) and the ascorbic acid level decreased (from 209.3 mg to 110.0 mg per 100g of pulp), showing a significant (p£0.05) inverse linear correlation (r=-0.98). The descriptors terms for mangoes were: characteristic flavor, characteristic aroma, sourness, astringency, yellow coloration of pulp, sweetness and succulence. The sensory profile presented significant improvement during ripening. All sensory attributes increased significantly (p£0.05) except sourness and astringency, wich decreased during the ripening of mangoes.

Estudo de bactérias do gênero Gluconobacter: isolamento, purificação, identificação fenotípica e molecular

A importância do estudo de bactérias acéticas, em especial as do gênero Gluconobacter, está baseada em suas aplicações industriais, pois estas possuem a capacidade de bioconversão de sorbitol a sorbose, viabilizando o processo de produção de vitamina C. O estudo envolveu coletas de amostras em indústrias de refrigerante, flores, frutos e mel, seguidas de purificação, identificação fenotípica e identificação molecular, com a utilização de iniciador definido a partir de consulta ao Nucleotide Sequence Database. Preservaram-se as linhagens identificadas como membros da família Acetobacteriaceae, gênero Gluconobacter. Foi isolado um total de 110 linhagens dos substratos: Pyrostegia venusta (Cipó de São João), mel, Vitis vinifera (uva), Pyrus communis (pêra), Malus sp. (maçã) e de duas amostras de refrigerantes envasados em embalagens de PET de 2 L. Deste total, 57 linhagens foram recuperadas em meio MYP (manitol, extrato de levedura, peptona), 12 em meio YGM (glicose, manitol, extrato de levedura, etanol, ácido acético), 41 em meio de enriquecimento e, posteriormente, em meio GYC (glicose, extrato de levedura e carbonato de cálcio). Obtiveram-se 68 linhagens identificadas como bastonetes Gram negativos. Destas, 31 foram caracterizadas bioquimicamente como pertencentes à família Acetobacteriaceae por serem catalase positivas, oxidase negativas e produtoras de ácido a partir de glicose. A caracterização dessas linhagens foi complementada com os testes bioquímicos: liquefação da gelatina, redução de nitrato, formação de indol e H2S e oxidação de etanol a ácido acético. Métodos moleculares foram aplicados para identificação do gênero Gluconobacter. Finalmente, oito linhagens foram caracterizadas como pertencentes ao gênero Gluconobacter. As linhagens encontram-se depositadas em coleção de cultura do laboratório de Microbiologia do Departamento de Biologia da UNESP, campus de Assis, estocadas em extrato de malte 20 a -196 ºC.