A fructan: fructan fructosyltransferase activity from Lolium rigidum

Fructan:fructan fructosyltransferase (FFT) activity was purified about 300-fold from leaves of Lolium rigidura Gaudin by a combination of affinity chromatography, gel filtration, anion exchange and isoelectric focusing. The FFT activity was free of sucrose:sucrose fructosyltransferase and invertase activities. It had an apparent pI of 4.7 as determined by isoelectric focusing, and a molecular mass of about 50000 (gel filtration). The FFT activity utilized the trisaccharides 1-kestose and 6(G)-kestose as sole substrates, but was not able to use 6-kestose as sole substrate. The FFT activity was not saturated when assayed at concentrations of 1-kestose, 6(G)-kestose or (1,1)-kestotetraose of up to 400 mM The rate of reaction of the FFT activity was most rapid when assayed with 1-kestose and was less rapid when assayed with 6(G)-kestose, (1,1)-kestotetraose or (1,1,1)-kestopentaose. The FFT activity when assayed at a relatively high concentration of enzyme activity (approximately equivalent to about half the activity in crude extracts per gram fresh mass) did not synthesize fructan of degree of polymerization > 6, even during extended assays of up to 10 h. When assayed with a combination of 1-kestose and uniformly labelled [C-14]sucrose as substrates, the major reaction was the transfer of a fructosyl residue from 1-kestose to sucrose resulting in the re-synthesis of 1-kestose. Tetrasaccharide and 6(G)-kestose were also synthesized. When assayed with 6(G)-kestose and [C-14]sucrose as substrates, the major reaction of the FFT activity was the synthesis of tetrasaccharide. However, some synthesis of 1-kestose and re-synthesis of 6(G)-kestose also occurred. When 6, kestose was the sole substrate for the FFT activity, synthesis of tetrasaccharide was 2.7 to 3.4-fold slower than when 1-kestose was used as the sole substrate. Owing to differences in the fructan:sucrose fructosyltransferase activity of the FFT with each of the trisaccharides, net synthesis of tetrasaccharide by the FFT was altered significantly in the presence of sucrose. The magnitude of this effect depended on the concentration of the trisaccharides. In the presence of sucrose, 6(G)-kestose could be a substrate of equivalent importance to 1-kestose for synthesis of tetrasaccharide.

Association between glutathione S-transferase polymorphisms and triglycerides and HDL-cholesterol

Objective: Null genotypes of glutathione S-transferase (GSTs) exhibit absence of enzymatic activity and are hypothesized to modulate an increased risk of developing cardiovascular disease. The aim of this study was to identify the potential association between GSTM1 and GSTT1 deleted polymorphisms with cardiovascular risk factors and coronary atherosclerosis in two independent urban populations. Methods and results: Genotype distribution of GSTM1 and GSTT1 deleted polymorphism were examined in a sample of 1577 individuals from the general population and a replication sample of 871 individuals submitted to coronary angiography. Triglycerides, HDL-cholesterol and the triglycerides/HDL ratio were significantly associated with a double-deleted genotype in individuals from the general population. These findings were replicated in a second, independent, population of individuals submitted to coronary angiography. In addition, coronary artery disease severity was also associated with GSTs genotypes and the risk conferred from GSTs genotype was mainly due to triglycerides/HDL ratio information. Conclusions: The data suggest that the presence of a double deletion genotypes of the GSTM1 and GSTT1 genes is associated with hypertriglyceridemia and low HDL-cholesterol levels in humans. These novel findings may provide a new unexplored link between lipid metabolism and GST homeostasis. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Granulocyte-Colony Stimulating Factor (G-CSF) induces mechanical hyperalgesia via spinal activation of MAP kinases and PI(3)K in mice

Granulocyte-colony stimulating factor (G-CSF) is a current pharmacological approach to increase peripheral neutrophil counts after anti-tumor therapies. Pain is most relevant side effect of G-CSF in healthy volunteers and cancer patients. Therefore, the mechanisms of G-CSF-induced hyperalgesia were investigated focusing on the role of spinal mitogen-activated protein (MAP) kinases ERK (extracellular signal-regulated kinase). JNK (Jun N-terminal Kinase) and p38, and PI(3)K (phosphatidylinositol 3-kinase). G-CSF induced dose (30-300 ng/paw)-dependent mechanical hyperalgesia, which was inhibited by local post-treatment with morphine. This effect of morphine was reversed by naloxone (opioid receptor antagonist). Furthermore, G-CSF-induced hyperalgesia was inhibited in a dose-dependent manner by intrathecal pre-treatment with ERK (PD98059), JNK (SB600125), p38 (SB202190) or PI(3)K (wortmanin) inhibitors. The co-treatment with MAP kinase and PI(3)K inhibitors, at doses that were ineffective as single treatment, significantly inhibited G-CSF-induced hyperalgesia. Concluding, in addition to systemic opioids, peripheral opioids as well as spinal treatment with MAP kinases and PI(3)K inhibitors also reduce G-CSF-induced pain. (C) 2011 Elsevier Inc. All rights reserved.

Farnesoic acid O-methyl transferase (FAMeT) isoforms: Conserved traits and gene expression patterns related to caste differentiation in the stingless bee, Melipona scutellaris

Farnesoic acid O-methyl transferase (FAMeT) is the enzyme that catalyzes the formation of methyl farnesoate (MF) from farnesoic acid (FA) in the biosynthetic pathway of juvenile hormone (JH). This work reports the cloning, sequencing, and expression of FAMeT gene from the stingless bee Melipona scutellaris (MsFAMeT). The MsFAMeT in silica analysis showed that greatest sequence similarity is found in Apis mellifera and other insects, while relatively less similarity is shown in crustaceans. Evidence of alternative splicing of a 27 nucleotide (nt) microexon explains the presence of the detected isoforms, 1 and 2. The expression analysis of the two isoforms showed a marked difference when castes were compared, suggesting that they could be involved differently in the JH metabolism in M. scutellaris, providing new insights for the comprehension of female plasticity.

Impact of PI and NNRTI HAART-based therapy on oral lesions of Brazilian HIV-infected patients

Background: The incidence of oral lesions related to human immunodeficiency virus (HIV) infection have been investigated after treatment with highly active antiretroviral therapy (HAART) including protease inhibitors (PI) but no data are available on the effect of non-nucleoside reverse transcriptase inhibitor (NNRTI)-based therapy on incidence of acquired immunodeficiency syndrome (AIDS) oral manifestations or impact of HAART on oral manifestations of HIV infection in Brazil. The aim of this study was to describe the effects of anti-HIV therapy on the incidence of oral lesions during 17 years of AIDS epidemics in a Brazilian population. Methods: From 1989 to 2006, we collected data from 1595 consecutive HIV patients at the Special Care Dentistry Center, Sao Paulo, Brazil. We compared the effect of PI- and NNRTI-based antiretroviral therapy (ARVT) on the annual incidence of Kaposi sarcoma (KS), oral candidiasis (OC) and hairy leukoplakia (HL). The chi-squared test was used to test the association between oral lesions and therapeutic regimen (P < 0.05). Results: None of patients on ARVT presented with KS. Patients who used (nucleoside reverse transcriptase inhibitors) NRTI + PI were 0.9 times as likely to present with HL as those who used NRTI + NNRTI. This finding, however, was not statistically significant (P = 0.5). The relative risk for OC was 0.8 in patients with PI-based HAART. The increased risk among those on PIs was statistically significant (P = 0.004). Conclusions: The superiority of NNRTI regimens in decreasing OC incidence is consistent with current therapeutic guidelines which recommend NNRTI-based therapy as the treatment of choice for initial ARVT.

Polymorphisms at the glutathione S-transferase GSTM1, GSTT1 and GSTP1 loci: risk of ovarian cancer by histological subtype

The phase II glutathione S-transferases (GSTs) GSTT1, GSTM1 and GSTP1 catalyse glutathione-mediated reduction of exogenous and endogenous electrophiles. These GSTs have broad and overlapping substrate specificities and it has been hypothesized that allelic variants associated with less effective detoxification of potential carcinogens may confer an increased susceptibility to cancer. To assess the role of GST gene variants in ovarian cancer development, we screened 285 epithelial ovarian cancer cases and 299 unaffected controls for the GSTT1 deletion (null) variant, the GSTM1 deletion (null) variant and the GSTP1 codon 104 A-->G Ile-->Val amino acid substitution variant, The frequencies of the GSTT1, GSTM1 and GSTP1 polymorphic variants did not vary with tumour behaviour (low malignant potential or invasive) or p53 immunohistochemical status. There was a suggestion that ovarian cancers of the endometrioid or clear cell histological subtype had a higher frequency of the GSTT1 and GSTM1 deletion genotype than other histological subgroups. The GSTT1, GSTM1 and GSTP1 genotype distributions did not differ significantly between unaffected controls and ovarian cancer cases (overall or invasive cancers only). However, the GSTM1 null genotype was associated with increased risk of endometrioid/clear cell invasive cancer [age-adjusted OR (95% CI) = 2.04 (1.01-4.09), P = 0.05], suggesting that deletion of GSTM1 may increase the risk of ovarian cancer of these histological subtypes specifically. This marginally significant finding will require verification by independent studies.

Use of heterologously-expressed cytochomre P450 and glutathione transferase enzymes in toxicity assays

Our groups have had a long-term interest in utilizing bacterial systems in the characterization of bioactivation and detoxication reactions catalyzed by cytochrome P450 (P450) and glutathione transferase (GST) enzymes. Bacterial systems remain the first choice for initial screens with new chemicals and have advantages, including high-throughput capability. Most human P450s of interest in toxicology have been readily expressed in Escherichia coli with only minor sequence modification. These enzymes can be readily purified and used in assays of activation of chemicals. Bicistronic systems have been developed in order to provide the auxiliary NADPH-P450 reductase. Alternative systems involve these enzymes expressed together within bacteria. In one approach, a lac selection system is used with E. coli and has been applied to the characterization of inhibitors of P450s 1A2 and 1131, as well as in basic studies involving random mutagenesis. Another approach utilizes induction of the SOS (umu) response in Salmonella typhimurium, and systems have now been developed with human P450s 1A1, 1A2, 1B1, 2C9, 2D6, 2E1, and 3A4, which have been used to report responses from heterocyclic amines. S. typhimurium his reporter systems have also been used with GSTs, first to demonstrate the role of rat GST 5-5 in the activation of dihalomethanes. These systems have been used to compare these GSTs with regard to activation of dihaloalkanes and potential toxicity. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.

O caso da arte gr??fica Waj??pi e o embate contra a explora????o comercial inadequada: limites e potencialidades das pol??ticas p??blicas de prote????o ao patrim??nio imaterial

O presente estudo tem por finalidade discutir os limites e potencialidades dos instrumentos de pol??ticas p??blicas de prote????o ao patrim??nio imaterial, ?? luz das experi??ncias constitu??das em um caso concreto, qual seja, o registro da arte Kusiwa. Em 2003, a arte Kusiwa dos Waj??pi, no Amap?? recebeu o reconhecimento como Obra-Prima do Patrim??nio Oral e Imaterial da Humanidade, conferido pela UNESCO, um ano ap??s ter sido inscrita no Livro das Formas de Express??o e ter sido registrada pelo IPHAN. Entretanto, apesar dessas a????es de salvaguarda, esse patrim??nio imaterial vem sendo reiteradamente violado por meio do abuso comercial do uso n??o autorizado dos grafismos. O fato de as formas de produ????o e circula????o de conhecimento entre os coletivos ind??genas envolverem rela????es espec??ficas, que n??o se limitam ??s propostas atualmente dispon??veis na legisla????o sobre propriedade intelectual, confere um aspecto desafiador sobre a tem??tica. A partir do recorrente ass??dio da comercializa????o do Kusiwa, come??ou a haver um aprendizado institucional por parte do IPHAN e de diversos outros atores, principalmente quanto aos questionamentos envolvendo o alcance jur??dico de prote????o do patrim??nio dos bens registrados. Por tudo isso, o registro da arte Kusiwa foi o resultado de um processo mais amplo de conquistas de direitos e constru????o de cidadania, e gerou efeitos importantes n??o apenas para assegurar a transmiss??o intergeracional do conjunto de conhecimentos da cultura Waj??pi, mas, principalmente, porque a salvaguarda desse patrim??nio configurou-se como instrumento estrat??gico para o fortalecimento de a????es e lutas sociais que buscam assegurar a garantia de direitos culturais e at?? mesmo de direitos humanos de maneira ampla

O estresse ocupacional do professor do ensino superior: a relação entre os sintomas de estresse e a atividade docente em duas instituições de ensino superior da cidade de Teresina-pi

RESUMO: Com a globalização e as mudanças provenientes da mesma, o ser humano vem sofrendo com o estresse ao tentar se adaptar e acompanhar tais acontecimentos seja no âmbito familiar, social, afetivo e profissional. Para desempenhar as funções docentes do ensino superior, são exigidas diversas condições, e o professor sofre um desgaste físico e psicológico ao tentar suprir determinadas demandas, o que em muitos casos, ocasiona o estresse ocupacional. Com o propósito de aprofundar conhecimentos sobre a relação existente entre o estresse e a atividade docente, fizemos o seguinte questionamento: quais os sintomas de estresse apresentados em professores do ensino superior? Com Objetivo geral de estudar os sintomas e níveis de estresse, relacionando-os à carreira docente do ensino superior, com as variáveis: regime de trabalho, vínculo institucional, idade, sexo, tempo de trabalho, tipo de IES, em uma população específica pertencente a duas Instituições de Ensino Superior -IES da cidade de Teresina, Estado do Piauí- Brasil, sendo uma pública e uma privada. Visando atingir os objetivos propostos, utilizamos da metodologia de caráter exploratório e descritivo, com abordagem quantitativa dos dados recorrendo à estatística descritiva. O instrumento de coleta de dados utilizado foi o “Inventário de Sintomas de Stress Lipp – ISSL” da autora Marilda Lipp (2000), aplicado em uma amostra de cem (100) professores, com idade entre 22 e 55 anos de idade. Mediante a análise dos dados, pode-se verificar que apesar das diferenças existentes entre IES públicas e privadas, a maioria dos resultados foram semelhantes. Em ambas as IES houve presença de estresse nos docentes, prevalecendo a fase de alerta, logo após a fase de resistência, em seguida a fase de quase-exaustão,Houve um predomínio de sintomas psicológicos, onde na IES privada se destacam: aumento súbito de motivação e entusiasmo súbito. E como sintomas físicos: boca seca e nó no estômago. Na IES públicas os sintomas psicológicos foram: entusiasmo súbito e aumento súbito de motivação. E físicos: tensão muscular e nó no estômago. Em relação as co-variáveis que apresentaram maior relação com o estresse pode-se apresentar na IES privada o sexo masculino e feminino possuem a mesma representatividade, a faixa etária de 31 a 39 anos, estado civil casado, tempo de serviço de 7 a 13 anos, jornada de trabalho integral e vínculo efetivo. Já na IES pública verifica-se relação com o sexo feminino, idade variando de 31 a 39 anos, estado civil solteiro, tempo de serviço de 7 a 13 anos, jornada de trabalho parcial e vínculo substituto. De acordo com o que foi analisado, percebe-se que os docentes do ensino superior, independente do tipo de IES ao qual estão vinculados, estão sujeitos ao estresse e que estes não devem ser responsabilizados pelo mesmo, nem por adoção de medidas preventivas, visto que toda a IES sofre as conseqüências, e tem condições de auxiliar o professor com situações preventivas, de controle a partir do conhecimento, interpretação e mudança de hábitos do professor no ambiente universitário. ABSTRACT: With globalization and its changes, the human being has suffered from stress while trying to adapt and monitor such events, within family, social, emotional or professional scope. To play as teacher in higher education, they are required various conditions and the teacher suffers physical and psychological wear trying to meet certain demands, which in many cases leads to occupational stress.Aiming to deepen knowledge about the relationship between stress and teaching activities, we made the following question: what are the symptoms of stress presented by higher education teachers? With the general objective of study the symptoms and the stress levels, linking them to the teaching career in higher education, using the following variables: working regime, institutional link, age, sex, working time, type of HEI, in a specific population belongs to two Higher Education Institutions HEI - of Teresina, Piauí, Brazil, one public and one private. Aiming to achieve the proposed objectives, we used a exploratory - descriptive study, with quantitative approach of data using graphs and tables. The data collection instrument used was the "Lipp Stress Symptoms Inventory - ISSL" by the author Marilda Lipp (2000), applied to a sample of a hundred (100) teachers, between 22 and 55 years of age. Through data analysis, despite the differences between public and private HEIs, we can see that the most results were similar. In both HEIs, the results revealed that the alert phase of stress had prevailed, followed by resistance and almost-exhaustion phases. A small number presented stress in exhaustion phase and another group did not present stress. There was a predominance of psychological symptoms, which in private HEIs stands out: increase of motivation and enthusiasm suddenly. And as physical symptoms: dry mouth and stomached. In public HEI, psychological symptoms were sudden enthusiasm and increse of motivation. And physical symptoms: muscle tension and stomached. In relation to the co-variables that showed a greater relationship with stress, in private HEIs males and females have the same representation, between 31-39 years, married, working time of 7-13 years, full working day and effective link. Already in public HEIs, female, between 31-39 years, unmarried, working time of 7-13 years, partial working day and substitute link had prevailed. According to analysis, we find that teachers in higher education, regardless of the HEI to which they are linked, are subject to stress. And they should not be blamed for the onset of stress, neither by adopting preventive measures, since all the HEI suffers the consequences, and it is able to help teachers with preventive measures based on the knowledge, interpretation and change habits of the teacher in the university environment.

Modificación del coeficiente peso/área del pié en relación con la agregación en Tarebia granifera

OBJETIVO: Analizar la dispersión de los ejemplares de una población de Tarebia granifera y su relación con la modificación de los parámetros morfométricos. METODOS: Se colectó por parcelas incluyendo dos tipos de substratos en condiciones opuestas de agregación y se comparó la abundancia relativa tanto entre muestreos, como entre sustratos. Se delimitaron 7 clases de alto a las cuales se les determinó el coeficiente peso/área del pié, el cual se correlacionó con la talla por clase. Se evaluó experimentalmente si los individuos con menor carga del pié tienen mayor persistencia en substratos emergidos, así como la interacción entre dos grupos de clases de alto. RESULTADOS: La diferencia de la abundancia entre los muestreos fue de t=5.874 (p<0.001) y de t=10.15 (p<0.001) entre el substrato rocoso emergido y el arenoso, sólo en condiciones de alta agregación, en las que la carga del pié de los individuos más grandes se reduce con la talla (r=-0.98, p<0.05). Experimentalmente, la abundancia de los ejemplares de tallas opuestas está mutuamente limitada (r=-0.87, p<0.01), la altura del oleaje limita la abundancia de los ejemplares menores (r=-0.94, p<0.001) siendo tolerada por los mayores (r=0.72, p<0.05). La carga promedio del pie disminuye con la altura del oleaje (r=-0.93, p<0.001). CONCLUSIONES: Con elevada agregación los ejemplares mayores emigran hacia substratos de difícil conquista por los menores, que están limitados por la acción mecánica del medio. La disminución de la carga del pie facilita la dispersión de los individuos.

Molinate quantification in environmental water by a glutathione-S-transferase based biosensor

A glutathione-S-transferase (GST)based biosensor was developed to quantify the thiocarbamate herbicide molinate in environmental water.The biosensor construction was based on GST immobilization onto a glassy carbon electrode via aminosilane–glutaraldehyde covalent attachment. The principle supporting the use of this biosensor consists of the GST inhibition process promoted by molinate. Differential pulse voltammetry was used to obtain a calibration curve for molinate concentration, ranging from 0.19 to 7.9 mgL -1 and presenting a detection limit of 0.064 mgL- 1. The developed biosensor is stable,and reusable during 15 days.The GST-based biosensor was successfully applied to quantify molinate in rice paddy field floodwater samples. The results achieved with the developed biosensor were in accordance with those obtained by high performance liquid chromatography. The proposed device is suitable for screening environmental water analysis and, since no sample preparation is required, it can be used in situ and in real-time measurements.

Microbiota fúngica dos condicionadores de ar nas unidades de terapia intensiva de Teresina, PI

Com intuito de identificar a microbiota fúngica em condicionadores de ar nas unidades de terapia intensiva de hospitais públicos e particulares de Teresina-PI, coletou-se material sólido de dez UTIs, isolando 33 espécies pertencentes às Moniliaceae e Dematiaceae, sendo primeira referência para o Piauí. Registrou-se elevada freqüência de Aspergillus niger Van Tieghem (60%); Aspergillus fumigatus Fres (50%); Trichoderma koningii Oudem (50%), Aspergillus flavus Link: Fr (40%). A validade da limpeza dos condicionadores de ar ultrapassou em todas as UTIs, a quantidade de unidades formadoras de colônia estava além do permitido pela Portaria 176/00 do Ministério da Saúde. É importante que os profissionais estejam munidos de equipamento de proteção individual, além de adotar medidas de controle de infecção hospitalar, sensibilizar para a existência de infecções fúngicas, melhorar ventilação de ar, possibilitando arejamento do ambiente e limpar periodicamente os condicionadores de ar, conscientizando os profissionais de saúde da importância destes fungos no ambiente hospitalar.

Análise funcional de uma quitinase da classe III (CgCHI3) e de uma glutationa S-transferase (CgGST), envolvidas na simbiose entre Casuarina glauca e Frankia

Fundação para a Ciência e Tecnologia - bolsa SFRH/ BD/41589/2007, cofinanciada pelo PIDDAC e pelo Fundo Social Europeu

Influência de sexo e energia sobre a taxa de crescimento, eficiência de utilização alimentar, taxa de uréia (U) e de proteína iodada do plasma (PI) em gado de corte

Três rações, com nível alto, médio e baixo de energia - tratamento T1 , T2 T3 - foram ministradas em confinamento, à vontade, a bovinos recém-desmamados, machos inteiros, castrados e fêmeas, durante dois períodos de 17 semanas. Os machos inteiros exibiram a maior habilidade de ganho de peso, e as novilhas a menor habilidade. O tratamento T1 proporcionou os melhores ganhos e conversões, e o T2 os inferiores. Verificou-se pequena superioridade no teor protéico de T3, ao que se atribuiu sua vantagem sobre T2 no 1º período, e aparente crescimento compensatório no 2º período. O teor de uréia foi superior nos machos inteiros, e semelhante nos outros animais, superior em T3 e semelhante em T1 e T2, bem como aumentou consistentemente durante o experimento. Aparentemente a uréia refletiu o fornecimento adequado de proteína e, mais ainda, de energia. O teor de proteína-iodada não mostrou diferença significativa, mas apenas tendência de se elevar com o decorrer do tempo. Não houve correlação entre os dois componentes do sangue. Concluiu-se que a eficiência de utilização alimentar foi: superior nos machos inteiros, proporcional à quantidade de energia ingerida (no caso do 1º período); favorecida pelo teor maior de proteína de T3 em relação a T2 no 19 período, e talvez por ganho compensatório no 2º período; refletida pelos níveis de uréia no plasma, mas não correlacionado com proteína- iodada.