Distribution of acid phosphatases in the hypopharyngeal glands from workers, queens and males of a Brazilian stingless bee Scaptotrigona postica Latreille: An ultrastructural cytochemical study

The present study reports the localization of acid phosphatase in the hypopharyngeal gland cells from workers (newly-emerged, nurse and forager), queens (newly-emerged and laying) and males (newly-emerged and mature for mating) of the Brazilian stingless bee, Scaptotrigona postica. The phosphatase activity varied in intensity and localization depending on the individual class, physiological age and the substrate used. In newly-emerged workers, the phosphatase-positive sites suggest the involvement of the enzyme with cellular differentiation that occurs in the presecretory phase, in nurse workers with protein synthesis and in forager workers with changes in cellular activity or glandular regression. In males mature for mating and laying queens, the positive sites are related to secretory activity, showing that the gland maintains some activity in spite of the regressive aspect. Of the substrates used, β-glycerophosphate gave the least specific localization.

Avaliação da população de neurônios mioentéricos NADPH-diaforase positivos do corpo do estômago de ratos com diabetes crônico induzido pela estreptozootocina

The aim of this study was to analyze the morphoquantitative features of the nitrergic myenteric neurons from the body of the stomach of diabetic rats. The body of the stomach of five normoglicemic rats and of five diabetic rats were prepared as whole-mounts stained by the histochemical technique of NADPH-diaphorase. Decreased body weight and increased daily ingestion of water, fast glicemia and glycated hemoglobin were verified in diabetic animals. According to the data obtained, significant difference in the density of nitrergic neurons was not observed between the two groups, but the areas of the neuronal cell body profiles in the diabetic rats were significantly larger. Results showed that the streptozotocin that induced diabetes does not accelerate the death of the nitrergic neurons, but increases the expression of these cells.

Biosynthetic origins of the isoprene units of 4-nerolidylcatechol in Potomorphe umbellata

The biosynthetic origins of the isoprene units of 4-nerolidylcatechol (1), the major constituent of Potomorphe umbellata, have been studied through feeding experiments with [14C]- and [13C]-glucose, and with precursors of the mevalonic acid and triose/pyruvate pathways, namely, [2- 14C]-mevalonolactone and [U-14C]-glyceraldehyde-3- phosphate, respectively. The pattern of incorporation of label from [1- 13C]-glucose into 1 was determined by quantitative 13C NMR spectroscopy. The labelling pattern revealed that the additive was specifically incorporated, and that the isoprene units of the sesquiterpenoid moiety of 4-nerolidylcatechol were derived from both the mevalonic acid and the triose/pyruvate pathways. The results indicate that both plastidic and cytoplasmic pathways are able to provide isopentenyl diphosphate units for the biosynthesis of 1. ©2005 Sociedade Brasileira de Química.

Eucalyptus ESTs associated with resistance to herbicide inhibitors of aromatic and branched-chain amino acid synthesis

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

Chorismate synthase: An attractive target for drug development against orphan diseases

The increase in incidence of infectious diseases worldwide, particularly in developing countries, is worrying. Each year, 14 million people are killed by infectious diseases, mainly HIV/AIDS, respiratory infections, malaria and tuberculosis. Despite the great burden in the poor countries, drug discovery to treat tropical diseases has come to a standstill. There is no interest by the pharmaceutical industry in drug development against the major diseases of the poor countries, since the financial return cannot be guaranteed. This has created an urgent need for new therapeutics to neglected diseases. A possible approach has been the exploitation of the inhibition of unique targets, vital to the pathogen such as the shikimate pathway enzymes, which are present in bacteria, fungi and apicomplexan parasites but are absent in mammals. The chorismate synthase (CS) catalyses the seventh step in this pathway, the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate. The strict requirement for a reduced flavin mononucleotide and the anti 1,4 elimination are both unusual aspects which make CS reaction unique among flavin-dependent enzymes, representing an important target for the chemotherapeutic agents development. In this review we present the main biochemical features of CS from bacterial and fungal sources and their difference from the apicomplexan CS. The CS mechanisms proposed are discussed and compared with structural data. The CS structures of some organisms are compared and their distinct features analyzed. Some known CS inhibitors are presented and the main characteristics are discussed. The structural and kinetics data reviewed here can be useful for the design of inhibitors. © 2007 Bentham Science Publishers Ltd.

Lipid peroxidation and antioxidant capacity of G6PD-deficient patients with A-(202G>A) mutation

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an enzymopathy in which reduced NADPH concentrations are not maintained, resulting in oxidative damage. We evaluated G6PD activity, oxidative stress levels and Trolox equivalent antioxidant capacity in individuals with the A-(202G>A) mutation for G6PD deficiency. Five hundred and forty-four peripheral blood samples were screened for G6PD deficiency; we also analyzed lipid peroxidation products measured as thiobarbituric acid reactive species and Trolox equivalent antioxidant capacity. Men with the A-(202G>A) mutation had lower G6PD activity than women with the same mutation. Individuals with the A-(202G>A) mutation also differed in mean Trolox equivalent antioxidant capacity values but not for thiobarbituric acid reactive species values. We concluded that A-(202G>A) mutation is associated with reduced G6PD activity and increased Trolox equivalent antioxidant capacity. ©FUNPEC-RP.

HDAC inhibition decreases XIST expression on female IVP bovine blastocysts

During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, although in vitro culture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In this study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development.We tested the hypothesis that by enhancing histone acetylation, TSA would increase the percentage of embryos achieving 16-cell stage, reducing percentage of embryos blocked at 8-cell stage, and interfere with XCI in IVF embryos. We noticed that after TSA treatment, acetylation levels in individual blastomeres of 8-16 cell embryos were increased twofold on treated embryos, and the samewas detected for blastocysts. Changes among blastomere levels within the same embryo were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by Hoechst staining, remained unaltered after TSA treatment. Then, we assessed XIST and G6PD expression in individual female bovine blastocysts by quantitative real-time PCR. Even though G6PD expression remained unaltered after TSA exposure, XIST expression was eightfold decreased, and we also detected a major decrease in the percentage of blastocysts expressing detectable XIST levels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrant XIST expression described for IVF embryos. © 2013 Society for Reproduction and Fertility.

Biochemical biomarkers in Scinax fuscovarius tadpoles exposed to a commercial formulation of the pesticide fipronil

One of the main pesticides used in the cultivation of sugarcane in São Paulo State, Brazil, is Regent®800WG, the main active compound of which is fipronil. Fipronil is a potent insecticide that eliminates pests, including insects resistant to pyrethroids, organophosphates (OP) and carbamates (CA). There is little known on the toxic effects of fipronil on non-target organisms, such as tadpoles of frogs. It is possible that this compound carries a high toxicity for these organisms, since the pesticide can be incorporated into aquatic environments during the rainy season, a time which coincides with the time of amphibian reproduction and the occurrence of tadpoles in the aquatic environment in this region. Thus, the pesticide could be contributing to the decline of amphibians in the northwest region of São Paulo state due to its wide use. This study aimed to test the influence of Regent®800WG on some biochemical systems of tadpoles (such as antioxidant defense systems) at different stages of development. The results of analysis from in vivo exposures demonstrated that only a few parameters in the groups exposed to fipronil responded to exposure to Regent®800WG, results which indicate that the pesticide instigates biochemical responses in tadpoles. Although catalase and glucose-6-phosphate dehydrogenase (G6PDH) were unchanged during the experiments, glutathione-S-transferase (GST) was inhibited in tadpoles, and the activity of glutathione reductase (GR) varied according to the exposure period and pesticide concentration. This data demonstrated the influence of the fipronil formulation on the metabolism of tadpoles, and showed that it can increase their susceptibility to environmental contaminants. © 2013 Elsevier Ltd. All rights reserved.

The Mode of Action of Recombinant Mycobacterium tuberculosis Shikimate Kinase: Kinetics and Thermodynamics Analyses

Tuberculosis remains as one of the main cause of mortality worldwide due to a single infectious agent, Mycobacterium tuberculosis. The aroK-encoded M. tuberculosis Shikimate Kinase (MtSK), shown to be essential for survival of bacilli, catalyzes the phosphoryl transfer from ATP to the carbon-3 hydroxyl group of shikimate (SKH), yielding shikimate-3-phosphate and ADP. Here we present purification to homogeneity, and oligomeric state determination of recombinant MtSK. Biochemical and biophysical data suggest that the chemical reaction catalyzed by monomeric MtSK follows a rapid-equilibrium random order of substrate binding, and ordered product release. Isothermal titration calorimetry (ITC) for binding of ligands to MtSK provided thermodynamic signatures of non-covalent interactions to each process. A comparison of steady-state kinetics parameters and equilibrium dissociation constant value determined by ITC showed that ATP binding does not increase the affinity of MtSK for SKH. We suggest that MtSK would more appropriately be described as an aroL-encoded type II shikimate kinase. Our manuscript also gives thermodynamic description of SKH binding to MtSK and data for the number of protons exchanged during this bimolecular interaction. The negative value for the change in constant pressure heat capacity (ΔCp) and molecular homology model building suggest a pronounced contribution of desolvation of non-polar groups upon binary complex formation. Thermodynamic parameters were deconvoluted into hydrophobic and vibrational contributions upon MtSK:SKH binary complex formation. Data for the number of protons exchanged during this bimolecular interaction are interpreted in light of a structural model to try to propose the likely amino acid side chains that are the proton donors to bulk solvent following MtSK:SKH complex formation. © 2013 Rosado et al.

Glicerol quinase de levedura de panificação

Pós-graduação em Alimentos e Nutrição - FCFAR

Alterações hematológicas e bioquímicas em uma população de Phrynops geoffroanus (Schweigger, 1812) em reposta a estressores ambientes

Pós-graduação em Genética - IBILCE

Alterações metabólicas de plantas de milho submetidas à aplicação de glyphosate e fosfito

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biomarcadores bioquímicos de contaminação em girinos de anfíbios anuros expostos a Regent®800WG (Fipronil)

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

Expressão gênica e viabilidade de folículos pré-antrais inclusos em fragmentos de córtex ovarianos cultivados in vitro de macaco-prego (Sapajus apella)

O objetivo desse trabalho foi analisar a ativação e o crescimento de folículos préantrais de Sapajus apella, submetidos a um sistema de cultivo in vitro em curto-prazo. Nesse sentido, dois experimentos foram realizados neste trabalho. Experimento I: exposição à fresco e à crioprotetores de biopsias de fragmentos do córtex ovariano. Ambos os fragmentos ovarianos foram submetidos à extração total de RNA e síntese de cDNA. Após a amplificação do cDNA por PCR em tempo-real (RT-PCR), os software GeNorm, bestkeeper e Normfinder foram usados para avaliar a estabilidade dos genes GAPDH (glyceraldehyde-2-phosphate dehydrogenase), HPRT1 (hypoxanthine phosphoribosyltransferase 1) e TBP (TATA-binding protein). Experimento II: o tecido do córtex ovariano de quatro fêmeas foi oletado e dividido em nove pedaços de 1 mm³. Um fragmento ovariano (grupo controle) foi imediatamente dividido em dois pedaços, os quais foram destinados para análise da viabilidade ou por RT-PCR. Os 8 fragmentos restantes foram individualmente cultivados in vitro em um meio constituído de TCM suplementado com 100 ng/mL EGF (T1), com adição de 10 μM de BME (T2), 100 ng/mL de BMP4 (T3), 25 IU de PMSG (T4), 10 μM de BME e 100 ng/mL de BMP4 (T5), 10 μM de BME, 25 IU de PMSG (T6), 100 ng/mL de BMP4, 25 IU de PMSG (T7) ou 10 μM de BME, 100 ng/mL de BMP4, 25 IU de PMSG (T8). Os resultados demonstraram que no tecido do córtex ovariano de S. apella, os genes HPRT1 e TBP foram os mais apropriados como genes de referência, podendo ser usados como parâmetro para normalizar dados em estudos futuros. Ao contrário, o GAPDH se apresentou como menos estável dos genes de referencia testados. Após o cultivo in vitro, todos os tratamentos alcançaram percentuais similares da viabilidade de folículos pré-antrais. O tecido ovariano cultivado na presença de GF+BME/BMP4/PMSG resultou no aumento da taxa de ativação e crescimento folicular, assim como no aumento da expressão de AMH, BMP15 e GDF9, genes conhecidos como indicadores específicos de desenvolvimento folicular. Dessa forma, o HPRT1 e TBP são os genes de referência mais estáveis, na exposição à crioprotetores, a fresco e no o cultivo de tecidos de córtex ovariano de S. apella. Os folículos pré-antrais são capazes de desenvolverem-se in vitro quando cultivados em meio suplementado com PMSG, BME e BMP4. A viabilidade folicular, entretanto, permaneceu independentemente do meio de cultivo in vitro e o uso de fatores de crescimento, como marcadores de desenvolvimento folicular, foi crucial para identificar o melhor meio de cultivo in vitro.

Influence of salts on the coexistence curve and protein partitioning in nonionic aqueous two-phase micellar systems

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