Avaliação do estado nutricional relativo ao selênio e da expressão gênica de selenoproteínas em pacientes com aterosclerose tratados com estatinas

The aim of this study was to determine the effects of the use of rosuvastatin in patients with atherosclerosis, in relation to blood parameters of selenium and selenoproteins, and also observe possible changes in gene expression of selenoproteins in these patients. The sample consisted of 27 adult and elderly patients with a clinical diagnosis of coronary artery disease undergoing angioplasty, treated at Natal Hospital Center hospital, Natal, RN. Patients were treated with rosuvastatin 10 mg/day during four months. Anthropometric variables such as body mass index (BMI) and Waist circumference (WC) were measured before and after treatment, as well as lipid profile, blood glucose and liver enzymes (AST and ALT). The diet of the patients was also analyzed using 24-hour diet recall. We analyzed the concentrations of selenium in plasma and erythrocytes, and also the activity of Glutathione Peroxidase and gene expression by Real Time PCR of selenoproteins GPx1, SelP1 and SelN1. Patients had mean age of 61.0 ± 9.4 years, 59.3% were men and 40.7% were women. After four months of treatment there was significant reduction of CA and, according to BMI, most were overweight. The intake of macronutrients, cholesterol, polyunsaturated fatty acids, monounsaturated and saturated was adequate, but the energy and fiber intake was below the recommendations. Regarding the selenium intake was observed a high prevalence of inadequacy. As expected, after treatment with rosuvastatin, a significant reduction in total cholesterol, LDL and glucose, which was not observed for HDL. Selenium concentrations in plasma and erythrocytes showed no changes, keeping within the established cutoffs. We observed a significant increase in GPx enzyme activity and mRNA expression of GPX1 and SEPN1, but not for gene SEPP1. Thus, it was found that treatment with rosuvastatin did not reduce the expression of selenoproteins. More studies are needed to clarify the effects of rosuvastatin on gene expression of selenoproteins in patients with atherosclerosis

Extração, caracterização e atividades biológicas de proteínas da espécie cnidoscolus urens (L.) Arthur

The extraction, chemical and structural characterization of a wide variety of compounds derived from plants has been a major source of bioactive molecules. Several proteases have been isolated in the plant kingdom, with numerous pharmacological and biotechnological applications. Among the proteases isolated from plants, are the fibrinogenolytic, with relevant application in the treatment of disorders in the coagulation cascade, in addition to potential use as a tool in clinical laboratories. In this study, in addition to evaluating the effects of the protein extract of Cnidoscolus urens (L.) Arthur (Euphorbiaceae) in the coagulation cascade also investigates the presence of antimicrobial activity and characterizes the proteolytic activity detected in this extract, aiming to determine their potential pharmacological and biotechnological application. In this way, crude protein extracts obtained from the leaves of C. urens in Tris-HCl 0.05M, NaCl 0.15M, pH 7.5, were precipitated in different concentrations of acetone, and assessed for the presence of proteolytic activity in azocaseína and fibrinogen. The most active fraction (F1.0) in these tests was chosen for assessment of biological activity and biochemical characterization. The Aα chain and Bβ of fibrinogen were completely cleaved at a concentration of 0.18 μg/μL of protein fraction in 4 minutes. Fibrinogenolytic activity presented total inhibition in the presence of E-64 and partial in the presence of EDTA. The fraction demonstrated coagulant activity in plasm and reduced the APTT, demonstrating acting on the factors coagulation of the intrinsic pathway and common, not exerting effects on the PT. Fibrinolytic activity on plasma clot was detected only in SDS-PAGE in high concentrations of fraction, and there were no defibrinating. Although several proteases isolated from plants and venomous animals are classically toxic, the fraction F1.0 of C. urens not expressed hemorrhagic nor hemolytic activities. Fraction F1.0 also showed no antimicrobial activity. In proteolytic activity on the azocasein, the optimal pH was 5.0 and optimum temperature of 60ºC. The enzyme activity has been shown to be sensitive to the presence of salts tested, with inhibition for all compounds. The surfactant triton did not influence the enzyme activity, but the tween-20 and SDS inhibited the activity. In the presence of reducing agents increase in enzyme activity occurred, a typical feature of enzymes belonging to the class of cysteine proteases. Several bands with proteolytic activity were detected in zymogram, in the region of high-molecular-weight, which were inhibited by E-64. In this study, we found that C. urens presents in its constitution cysteine proteases with fibrinogenolytic and procoagulant activity, which may be isolated, with potential application in treatment of bleeding disorders, thrombolytic and clinical laboratory

Avaliação da composição química, atividade antioxidante, antibacteriana, antinoceptiva, antiinflamatória e toxicidade do extrato metanólico e frações de folhas de Spondias sp. (Anacardiaceae)

Spondias sp. (Anacardiaceae), popularly known as cajá-umbu, is an endemic plant from Northeastern Brazil, where their leaves are widely used in folk medicine to treat inflammatory processes, while their fruits have a great agro industrial potential. This study was designed to evaluate hepatoprotective, antinociceptive, antioxidant, antimicrobial and anti-inflammatory properties, as well as the acute toxicity and repeated dose 28, using a methanolic extract (MES), a fraction rich in flavonoids (FRF) and a precipitate from Spondias sp.leaves. The antioxidant activity of them was valued to evaluate their free radical scavenger capacity by DPPH test, whereas MES and FRF were used to evaluate while the preventive action on carbon tetrachloride (CCl4)-induced hepatotoxicity. Seven groups (n=5) of female Wistar rats were used as follows: control group, CCl4-intoxicated group treated with EMS (500 mg/kg) for 7 days, three CCl4-intoxicated groups treated with FRF (25, 50 and 75 mg/kg) for 7 days and the CCl4-intoxicated group treated with Legalon ® (silimarina; (phytotherapeutic reference) (50 mg/kg; 7 days). MES and FRF showed a protective action against liver injury induced by CCl4, being observed a significant reduction of serum enzyme activity marker of liver damage (alanine transaminase and aspartate transaminase). On the other hand, the lipid peroxidation (SRAT) decrease, as well as the increase of glutathione content and enzyme activity of antioxidant defense system (SOD, CAT, GPx) toward near normal values indicated the ability of EMS to restore the oxidative imbalance induced by CCl4. The histological analysis confirmed the hepatoprotection, compared to degenerative changes in CCl4-treated group. This hepatoprotetor effect was similar to that shown by Legalon®. The in vitro high antioxidant capacity of extract (93.16 ± 1.00%) showed analogous results to those obtained by Carduus marianus BHT (reference standard). This fact explains the obtained results in vivo. Although no antimicrobial activity was detected, EMS and FRF promoted the antinociceptive effect induced in the second phase by the intraplantar formalin test, evidencing the anti-inflammatory action; confirmed by the carrageenan-induced peritonitis model. The evaluation of the mechanical allodynia (CFA a 80%) demonstrated the involvement of the Spondias sp. chemical composition in the anti-inflammatory activity toward the acute processes. The acute exposure and repeated dose during 28 days did not produce significant changes in the parameters that evaluate toxicity. Together the experimental results reveal, that Spondias sp. leaf extracts have a promising potential in pharmaceutical area, and due to its non-toxic condition present efficiency and security

Avaliação do status antioxidante, expressão gênica e polimorfismos dos genes SOD1, SOD2 e GPx1 em crianças, adolescentes e adultos jovens com diabetes tipo 1

Studies report that the pathophysiological mechanism of diabetes complications is associated with increased production of Reactive Oxygen Species (ROS)-induced by hyperglycemia and changes in the capacity the antioxidant defense system. In this sense, the aim of this study was to evaluate changes in the capacity of antioxidant defense system, by evaluating antioxidant status, gene expression and polymorphisms in the genes of GPx1, SOD1 and SOD2 in children, adolescents and young adults with type 1 diabetes. We studied 101 individuals with type 1 diabetes (T1D) and 106 normoglycemic individuals (NG) aged between 6 and 20 years. Individuals with type 1 diabetes were evaluated as a whole group and subdivided according to glycemic control in DM1G good glycemic control and DM1P poor glycemic control. Glycemic and metabolic control was evaluate by serum glucose, glycated hemoglobin, triglycerides, total cholesterol and fractions (HDL and LDL). Renal function was assessed by measurement of serum urea and creatinine and albumin-to-creatinine ratio (ACR) in spot urine. Antioxidant status was evaluate by content of reduced glutathione (GSH) in whole blood and the activity of erythrocyte enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD). We also analyzed gene expression and gene polymorphisms of GPx1 (rs1050450), SOD1 (rs17881135) and SOD2 (rs4880) by the technique of real-time PCR (Taqman®). Most individuals with DM1 (70.3%) had poor glycemic control (glycated hemoglobin> 8%). Regarding the lipid profile, individuals with type 1 diabetes had significantly elevated total cholesterol (p <0.001) and LDL (p <0.000) compared to NG; for triglycerides only DM1NC group showed significant increase compared to NG. There was an increase in serum urea and RAC of individuals with DM1 compared to NG. Nine individuals with type 1 diabetes showed microalbuminuria (ACR> 30 mg / mg). There was a decrease in GSH content (p = 0.006) and increased erythrocyte GPx activity (p <0.001) and SOD (p <0.001) in DM1 group compared to NG. There was no significant difference in the expression of GPx1 (p = 0.305), SOD1 (.365) and SOD2 (0.385) between NG and DM1. The allele and genotype frequencies of the polymorphisms studied showed no statistically significant difference between the groups DM1 and NG. However, the GPx1 polymorphism showed the influence of erythrocyte enzyme activity. There was a decrease in GPx activity in individuals with type 1 diabetes who had a polymorphic variant T (p = 0.012). DM1 patients with the polymorphic variant G (AG + GG) for polymorphism of SOD2 (rs4880) showed an increase in the RAC (p <0.05). The combined data suggest that glucose control seems to be the predominant factor for the emergence of changes in lipid profile, renal function and antioxidant system, but the presence of the polymorphisms studied may partly contribute to the onset of complications

Microbial alterations of the soil influenced by induced compaction

A compactação é um dos fatores mais agravantes para a qualidade do solo, porém o seu efeito na comunidade e atividade enzimática microbiana não tem sido suficientemente estudado. Seis níveis de compactação foram obtidos pela passagem de tratores com diferentes pesos em um Latossolo Vermelho, e a densidade final foi medida. Amostras de solo foram coletadas nas profundidades de 0-10 e 10-20 cm, após a colheita do milho. O efeito da compactação foi evidente em todos os parâmetros estudados, mas nem sempre foi significativo. A contagem das bactérias totais reduziu significativamente em 22-30 %, e a das nitrificantes, em 38-41 %, no solo com maior densidade em relação ao controle. Contudo, a população de fungos aumentou de 55 a 86 %, e a das bactérias desnitrificantes, de 49 a 53 %. A atividade da desidrogenase diminuiu de 20 a 34 %; a da urease, de 44 a 46 %; e a da fosfatase, de 26 a 28 %. O conteúdo de matéria orgânica e o pH do solo diminuíram na camada 0-0,10 em relação à de 0,10-0,20 m e influíram possivelmente na redução das contagens microbianas exceto das bactérias desnitrificantes, e na atividade das enzimas, menos a da urease. Esses resultados indicam que a compactação do solo teve influência na comunidade de microrganismos aeróbios e na sua atividade. Esse efeito pode alterar a ciclagem de nutrientes e diminuir a produção da planta.

Preservation of minimally processed aurora-1 peaches using additives

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

Utilização de dietas úmidas e de rações e água de bebida com edulcorante para leitões desmamados aos 21 dias de idade e efeitos sobre o desenvolvimento histológico e enzimático intestinal

Um experimento foi realizado com leitões desmamados visando avaliar os efeitos de oito tratamentos, correspondentes à combinação dos fatores, apresentações de ração (seca e úmida), tipos de ração (sem e com edulcorante) e tipos de água de consumo (sem e com edulcorante) sobre o desenvolvimento morfológico e enzimático intestinal. Foram avaliadas a altura das vilosidades (V), a profundidade das criptas (C) e a relação altura das vilosidades e profundidade das criptas (V:C) do duodeno e do jejuno, o conteúdo de proteína na mucosa do jejuno, e as atividades enzimáticas (UA) da maltase, sacarase e dipeptidase. Utilizaram-se 32 leitões machos castrados, Large White x Landrace, desmamados aos 21 dias, submetidos aos tratamentos durante oito dias. O delineamento experimental foi em blocos casualizados, com oito tratamentos, fatorial 2 x 2 x 2, com quatro repetições. Foram observadas diferenças para a altura das vilosidades do duodeno para os grupos tratados com dieta úmida e ração com edulcorante e para a relação V:C do duodeno para os animais que receberam ração com edulcorante. Houve diferença para a UA da sacarase e da maltase para os grupos que receberam água sem edulcorante. As características histológicas foram favorecidas pelas dietas úmidas, o que pode sugerir que esta apresentação foi menos agressiva ao trato digestivo. Para os demais fatores os resultados contrariaram as expectativas de que o tratamento água com edulcorante melhoraria a qualidade do epitélio intestinal. A fase crítica pela qual os leitões foram submetidos, possivelmente, comprometeu os resultados.

Fenólicos totais, polifenoloxidade e coloração em abacate 'Hass' submetido a radiação UV-C

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mechanism and control of Genipa americana seed germination

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Funcionalidade dos extratos fenólicos obtidos pelo cultivo semi-sólido de resíduos de abacaxi (Ananas comusus L.) e goiaba (Psidium guajava L.)

Solid substrate cultivation (SSC) has become an efficient alternative towards rational use of agro industrial wastes and production of value-added products, mainly in developing countries. This work presents the production and functional application results of phenolic extracts obtained by solid substrate cultivation of pineapple (Ananas comosus L.) and guava (Psidium guajava L.) residues associated to soy flour and bioprocessed by Rhizopus oligosporus fungus. Two experimental groups were tested: (1) 9g of fruit residue and 1g of soy flour (A9 or G9); (2) 5g of fruit residue and 5g of soy flour (A5 or G5). After SSC, 100ml of distilled water was added to each Erlenmeyer flask containing 10g of bioprocessed material in order to obtain the phenolic extracts. Samples were taken every two days for total phenolic concentration (TPC) and antioxidant capacity evaluation by DPPH test during 12-day cultivation. The 2-day and 10-d ay extracts were selected and concentrated by ebullition until 1/10 of original volume was reached. After that, both non-concentrated and concentrated extracts were evaluated for their antimicrobial activity against Staphylococcus aureus and Salmonella enterica and a-amylase inhibitory capacity. It was observed an inverse relationship between total phenolic concentration (TPC) and antioxidant capacity during the cultivation. Besides that, the concentrated pineapple samples after two days were able to inhibit both pathogens tested, especially S. aureus. Guava concentrated extracts after 2 days showed expressive inhibition against S. enterica, but negative results against S. aureus growth. When it comes to a-amylase inhibition, A9 extracts after 2 days, both concentrated or not, completely inhibited enzyme activity. Similar behavior was observed for G9 samples, but only for concentrated samples. It was shown that concentration by ebullition positively affected the enzymatic inhibition of G9 and A9 samples, but on the other side, decreased antiamylase activity of A5 and G5 samples

Recuperação e purificação de quitosanases usando adsorção em leito expandido com streamline DEAE com modelagem e simulação usando redes neurais

Expanded Bed Adsorption (EBA) is an integrative process that combines concepts of chromatography and fluidization of solids. The many parameters involved and their synergistic effects complicate the optimization of the process. Fortunately, some mathematical tools have been developed in order to guide the investigation of the EBA system. In this work the application of experimental design, phenomenological modeling and artificial neural networks (ANN) in understanding chitosanases adsorption on ion exchange resin Streamline® DEAE have been investigated. The strain Paenibacillus ehimensis NRRL B-23118 was used for chitosanase production. EBA experiments were carried out using a column of 2.6 cm inner diameter with 30.0 cm in height that was coupled to a peristaltic pump. At the bottom of the column there was a distributor of glass beads having a height of 3.0 cm. Assays for residence time distribution (RTD) revelead a high degree of mixing, however, the Richardson-Zaki coefficients showed that the column was on the threshold of stability. Isotherm models fitted the adsorption equilibrium data in the presence of lyotropic salts. The results of experiment design indicated that the ionic strength and superficial velocity are important to the recovery and purity of chitosanases. The molecular mass of the two chitosanases were approximately 23 kDa and 52 kDa as estimated by SDS-PAGE. The phenomenological modeling was aimed to describe the operations in batch and column chromatography. The simulations were performed in Microsoft Visual Studio. The kinetic rate constant model set to kinetic curves efficiently under conditions of initial enzyme activity 0.232, 0.142 e 0.079 UA/mL. The simulated breakthrough curves showed some differences with experimental data, especially regarding the slope. Sensitivity tests of the model on the surface velocity, axial dispersion and initial concentration showed agreement with the literature. The neural network was constructed in MATLAB and Neural Network Toolbox. The cross-validation was used to improve the ability of generalization. The parameters of ANN were improved to obtain the settings 6-6 (enzyme activity) and 9-6 (total protein), as well as tansig transfer function and Levenberg-Marquardt training algorithm. The neural Carlos Eduardo de Araújo Padilha dezembro/2013 9 networks simulations, including all the steps of cycle, showed good agreement with experimental data, with a correlation coefficient of approximately 0.974. The effects of input variables on profiles of the stages of loading, washing and elution were consistent with the literature

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

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evidence for conformational changes in the yeast deoxyhypusine hydroxylase Lia1 upon iron displacement from its active site

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

Recombinant expression of glycerol-3-phosphate dehydrogenase using the Pichia pastoris system

In the present study, the GPD2 gene from Saccharomyces cerevisiae, which codifies for the enzyme glycerol-3-phosphate dehydrogenase (GPDH), was cloned from the pPICZ-alpha expression vector and used with the purpose of inducing the extracellular expression of the glycerol-3-phosphate dehydrogenase under the control of the methanol-regulated AOX promoter. The presence of the GPD2 insert was confirmed by PCR analysis. Pichia pastoris X-33 (Mut(+)) was transformed with linearized plasmids by electroporation and transformants were selected on YPDS plates containing 100 mu g/mL of zeocin. Several clones were selected and the functionality of this enzyme obtained in a culture medium was assayed. Among the mutants tested, one exhibited 3.1 x 10(-2) U/mg of maximal activity. Maximal enzyme activity was achieved at 6 days of growth. Medium composition and pre-induction osmotic stress influenced protein production. Pre-induction osmotic stress (culturing cells in medium with either 0.35 M sodium chloride or 1.0 M sorbitol for 4h prior to induction) led to an increase in cell growth with sorbitol and resulted in a significant increase in GPDH productivity with sodium chloride in 24h of induction approximately fivefold greater than under standard conditions (without pre-induction). (C) 2010 Elsevier B.V. All rights reserved.

Efeito da temperatura de estocagem de leveduras de panificação sobre a atividade da glicerol-3-fosfato desidrogenase

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