PROLACTIN INDUCES MATURATION OF GLUCOSE SENSING MECHANISMS IN CULTURED NEONATAL RAT ISLETS

The effects of PRL treatment on insulin content and secretion, and Rb-86 and Ca-45 fluxes from neonatal rat islets maintained in culture for 7-9 days were studied. PRL treatment enhanced islet insulin content by 40% and enhanced early insulin secretion evoked by 16.7 mm glucose. Insulin release stimulated by oxotremorine-M, a muscarinic agonist, in the presence of glucose (8.3 or 16.7 mm) was unchanged by PRL treatment. However, PRL treatment potentiated phorbol 12,13-dibutyrate-stimulated insulin secretion in the presence of the above glucose concentrations. PRL treatment potentiated the reduction in Rb-86 efflux induced by glucose or tolbutamide and enhanced the increase in Rb-86 efflux evoked by diazoxide. PRL treatment slightly potentiated the increment in Ca-45 uptake induced by high concentrations of K+, but failed to affect the increment evoked by 16.7 mm glucose. Since glucose-induced Ca-45 uptake was not affected by PRL, we suggest that the enhancement in first phase insulin secretion evoked by glucose in the PRL-treated islets occurs at a step in the secretory process that may involve protein kinase-C. These data further support observations that PRL treatment increases islet sensitivity to glucose.

Dehydromonocrotaline induces cyclosporine A-insensitive mitochondrial permeability transition/cytochrome c release

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

Using Efinol (R) L during transportation of marbled hatchetfish, Carnegiella strigata (Gunther)

The objective of this experiment was to test the efficacy of a probiotic (Efinol (R) L) during transportation of marbled hatchetfish, Carnegiella strigata. Wild specimens were captured from a small stream and transported for 24 h in plastic fish boxes with a probiotic (10 mg L-1) and probiotic-free water. The boxes were sampled at 3. 12 and 24 h of transport. At the end of the experiment, the survival rate was close to.100%) in both treatments. Dissolved oxygen diminished with time in both treatments, but the probiotic group had significantly higher levels. Conductivity. pH and ammonia increased significantly during the transport. demonstrating higher levels in the probiotic-free group. Fish from both treatments presented very high net Na+ and K+ effluxes after 3 h of transport. At 24 h, net K+ effluxes in fish of the probiotic treatment reached values close to zero and a significantly lower Na+ efflux was observed. Cortisol levels in both treatments at 3 and 12 h were significantly higher than that in control samples. Higher body cortisol levels were observed in the probiotic-frec group than that in the probiotic group at 3 and 12 h. The results demonstrate that addition of a probiotic during fish transport improves water quality and leads to fish presenting a lower stress response intensity.

INGAP-PP up-regulates the expression of genes and proteins related to K-ATP(+) channels and ameliorates Ca2+ handling in cultured adult rat islets

Islet Neogenesis Associated Protein (INGAP) increases pancreatic beta-cell mass and potentiates glucose-induced insulin secretion. Here, we investigated the effects of the pentadecapeptide INGAP-PP in adult cultured rat islets upon the expression of proteins constitutive of the K-ATP(+) channel, Ca2+ handling, and insulin secretion. The islets were cultured in RPMI medium with or without INGAP-PP for four days. Thereafter, gene (RT-PCR) and protein expression (Western blotting) of Foxa2, SUR1 and Kir6.2, cytoplasmic Ca2+ ([Ca2+](i)), static and dynamic insulin secretion, and Rb-86 efflux were measured. INGAP-PP increased the expression levels of Kir6.2, SUR1 and Foxa2 genes, and SUR1 and Foxa2 proteins. INGAP-PP cultured islets released significantly more insulin in response to 40 mM KCl and 100 mu M tolbutamide. INGAP-PP shifted to the left the dose-response curve of insulin secretion to increasing concentrations of glucose (EC50 of 10.0 +/- 0.4 vs. 13.7 +/- 1.5 mM glucose of the controls). It also increased the first phase of insulin secretion elicited by either 22.2 mM glucose or 100 mu M tolbutamide and accelerated the velocity of glucose-induced reduction of Rb-86 efflux in perifused islets. These effects were accompanied by a significant increase in [Ca2+](i) and the maintenance of a considerable degree of [Ca2+](i) oscillations. These results confirm that the enhancing effect of INGAP-PP upon insulin release, elicited by different secretagogues, is due to an improvement of the secretory function in cultured islets. Such improvement is due, at least partly, to an increased K-ATP(+) channel protein expression and/or changing in the kinetic properties of these channels and augmented [Ca2+](i) response. Accordingly, INGAP-PP could potentially be used to maintain the functional integrity of cultured islets and eventually, for the prevention and treatment of diabetes. (C) 2008 Elsevier B.V. All rights reserved.

Detection of P. aeruginosa harboring bla(CTX-M-2), bla(GES-1) and bla(GES-5), bla(IMP-1) and bla(SPM-1) causing infections in Brazilian tertiary-care hospital

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

Long-term effect of prolactin treatment on glucose-induced insulin secretion in cultured neonatal rat islets

Insulin secretion and 45SCa2+ uptake and efflux were studied in neonatal rat islets maintained in culture for 7 or 19 days in the absence or presence of prolactin (PRL). Insulin secretion in response to glucose (G), leucine (Leu), arginine (Arg) and carbachol (Cch) was augmented after 7 and 19 days in culture, compared to basal secretion (G 2.8 mM), in both PRL- treated and control islets. However, the increase in insulin secretion induced by the above secretagogues was higher in islets cultured in the presence of PRL for 19 days. In PRL-treated islets, the 45Ca2+ content after a 5 min incubation in the presence of G, Leu, Arg and Cch was significantly higher than the control only in islets cultured for 19 days. Except with Arg, the 45Ca2+ uptake in PRL-treated islets after a 90 min incubation was also significantly higher than the control only in islets cultured for 19 days. Finally, Leu-induced alterations in the 45Ca2+ efflux were higher in PRL-treated than in control islets cultured for 7 or 19 days. In the absence of external Ca2+, the reduction in 45Ca2+ efflux induced by glucose was also significantly higher in PRL-treated than in control islets. This effect was slightly potentiated after 19 days in culture. These data further support the hypothesis that PRL treatment enhances maturation of the secretory mechanism in neonatal islets. This effect can be potentiated even more if the treatment is prolonged.

ZmPUMP encodes a fully functional monocot plant uncoupling mitochondrial protein whose affinity to fatty acid is increased with the introduction of a His pair at the second matrix loop

Uncoupling proteins (UCPs) are specialized mitochondrial transporter proteins that uncouple respiration from ATP synthesis. In this study, cDNA encoding maize uncoupling protein (ZmPUMP) was expressed in Escherichia coli and recombinant ZmPUMP reconstituted in liposomes. ZmPUMP activity was associated with a linoleic acid (LA)-mediated H+ efflux with Km of 56.36 ± 0.27 μM and Vmax of 66.9 μmol H+ min-1 (mg prot)-1. LA-mediated H+ fluxes were sensitive to ATP inhibition with Ki of 2.61 ± 0.36 mM (at pH 7.2), a value similar to those for dicot UCPs. ZmPUMP was also used to investigate the importance of a histidine pair present in the second matrix loop of mammalian UCP1 and absent in plant UCPs. ZmPUMP with introduced His pair (Lys155His and Ala157His) displayed a 1.55-fold increase in LA-affinity while its activity remained unchanged. Our data indicate conserved properties of plant UCPs and suggest an enhancing but not essential role of the histidine pair in proton transport mechanism. © 2006 Elsevier Inc. All rights reserved.

Evidence for the role of calcineurin in morphogenesis and calcium homeostasis during mycelium-to-yeast dimorphism of Paracoccidioides brasiliensis

Paracoccidioides brasiliensis is a dimorphic fungus that causes paracoccidioidomycosis, the most prevalent human deep mycosis in Latin America. The dimorphic transition from mycelium to yeast (M-Y) is triggered by a temperature shift from 25°C to 37°C and is critical for pathogenicity. Intracellular Ca 2+ levels increased in hyphae immediately after temperature-induced dimorphism. The chelation of Ca 2+ with extracellular (EGTA) or intracellular (BAPTA) calcium chelators inhibited temperature-induced dimorphism, whereas the addition of extracellular Ca 2+ accelerated dimorphism. The calcineurin inhibitor cyclosporine A (CsA), but not tacrolimus (FK506), effectively decreased cell growth, halted the M-Y transition that is associated with virulence, and caused aberrant growth morphologies for all forms of P. brasiliensis. The difference between CsA and FK506 was ascribed by the higher levels of cyclophilins contrasted to FKBPs, the intracellular drug targets required for calcineurin suppression. Chronic exposure to CsA abolished intracellular Ca 2+ homeostasis and decreased mRNA transcription of the CCH1 gene for the plasma membrane Ca 2+ channel in yeast-form cells. CsA had no detectable effect on multidrug resistance efflux pumps, while the effect of FK506 on rhodamine excretion was not correlated with the transition to yeast form. In this study, we present evidence that Ca 2+/calmodulin-dependent phosphatase calcineurin controls hyphal and yeast morphology, M-Y dimorphism, growth, and Ca 2+ homeostasis in P. brasiliensis and that CsA is an effective chemical block for thermodimorphism in this organism. The effects of calcineurin inhibitors on P. brasiliensis reinforce the therapeutic potential of these drugs in a combinatory approach with antifungal drugs to treat endemic paracoccidioidomycosis. Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Photodynamic inactivation of biofilm: Taking a lightly colored approach to stubborn infection

Microbial biofilms are responsible for a variety of microbial infections in different parts of the body, such as urinary tract infections, catheter infections, middle-ear infections, gingivitis, caries, periodontitis, orthopedic implants, and so on. The microbial biofilm cells have properties and gene expression patterns distinct from planktonic cells, including phenotypic variations in enzymic activity, cell wall composition and surface structure, which increase the resistance to antibiotics and other antimicrobial treatments. There is consequently an urgent need for new approaches to attack biofilm-associated microorganisms, and antimicrobial photodynamic therapy (aPDT) may be a promising candidate. aPDT involves the combination of a nontoxic dye and low-intensity visible light which, in the presence of oxygen, produces cytotoxic reactive oxygen species. It has been demonstrated that many biofilms are susceptible to aPDT, particularly in dental disease. This review will focus on aspects of aPDT that are designed to increase efficiency against biofilms modalities to enhance penetration of photosensitizer into biofilm, and a combination of aPDT with biofilm-disrupting agents. © 2013 Informa UK Ltd.

Partitioning of net primary production in Eucalyptus and Acacia stands and in mixed-species plantations: Two case-studies in contrasting tropical environments

The introduction of nitrogen fixing species (NFS) in fast-growing tree plantations is an alternative option to reduce fertilizer inputs. However, the success of mixed-species plantations depends on the balance between positive interactions among species (resulting from facilitation and/or complementarity) and the negative effects of interspecific competition.Using a carbon budget approach and coupling measurements of standing biomass, aboveground litterfall and soil CO2 efflux, we assessed the influence of replacing half of eucalypt trees by Acacia mangium on total belowground carbon flux (TBCF), net primary production (NPP) and its partitioning between above- and belowground growth at two tropical sites in Brazil (Itatinga) and in Congo (Kissoko) exhibiting contrasting climates, edaphic conditions and wood productions.Annual soil CO2 efflux (FS) was significantly lower in the acacia monocultures than in eucalypt monocultures and mixed-species stands at both sites. Annual FS was significantly lower at Itatinga compared to Kissoko for all stands while TBCF was significantly lower in the eucalypt stands only. In the eucalypt monocultures we found a significantly lower aboveground NPP (ANPP) and wood production (wood NPP) at Kissoko compared to Itatinga that was almost fully balanced by a significantly higher belowground NPP (BNPP), leading to similar NPP. Similarly, acacia monocultures exhibited significantly higher ANPP and wood NPP at Itatinga than at Kissoko. The mixed-species stands exhibited a significantly lower wood NPP and ANPP than the eucalypt monocultures at the Brazilian site while NPP of the mixture was not significantly different than the average NPP of the two monocultures. At the Congolese site, NPP of the mixture was significantly higher than the average NPP of the two monocultures. NPP was similar in the mixed-species stand and the eucalypt monoculture with a significantly lower partitioning of NPP to belowground production, leading to a one third higher wood biomass at harvest in the mixed-species stand.A positive effect of growing eucalypts with the nitrogen fixing acacia trees on stand wood production occurred at Kissoko but not at Itatinga. Mixed-species plantations with NFS can be advocated at sites where the productive gains resulting from nitrogen fixation are not compromised by other resource limitations. © 2012 Elsevier B.V.

Expressão da glicoproteína-p e da mrp1 em tecidos de cães portadores de leishmaniose visceral

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Transporte de CO2 no Sistema Solo-Planta-Atmosfera

Pós-graduação em Física - IGCE

Fluxos de CO2 do solo na Floresta Nacional de Caxiuanã, Pará, durante o experimento ESECAFLOR/LBA

O experimento ESECAFLOR/LBA foi conduzido na Floresta Nacional de Caxiuanã, Pará, e este artigo procura investigar os efeitos do estresse hídrico sobre a respiração do solo. Duas parcelas de 1 hectare foram instaladas em janeiro de 2002. Uma parcela (A) permaneceu em condições naturais e foi usada como controle, enquanto que na parcela de exclusão (B) foram instalados painéis plásticos para que aproximadamente 70% da precipitação fosse excluída. Os dados foram coletados mensalmente de janeiro a dezembro de 2005. Durante o ano de 2005 houve 2.211,6 mm de precipitação na ECFPn, ou seja 9,96% acima da média de 2.011,2 mm. As médias da umidade do solo foram de 15,6±9,2 e 9,5±3,4% nas parcelas A e B, respectivamente. As médias da temperatura do solo para as parcelas A e B foram de 25,6±0,4 e 25,7±0,5 ºC, respectivamente. As médias dos fluxos de CO₂ no solo nas parcelas A e B foram de 3,46±0,44 e 3,21±0,84 μmolCO₂ m^-2s^-1, respectivamente. Com a exclusão de parte da chuva na parcela B, houve uma redução de 7,23% nos fluxos de CO₂ no solo (0,25 μmolCO₂ m^-2s^-1), 39,1% na umidade do solo (6,1p.p.), e um aumento de 0,39% na temperatura do solo (0,1ºC). A umidade do solo na parcela B foi menor do que na parcela A, devido ao sistema de exclusão da chuva, no entanto no início do ano, devido a reformas que o mesmo passou esses valores estiveram próximos.

Variação temporal do efluxo de dióxido de carbono CO2 do solo em sistemas agroflorestais com palma de óleo (Elaeis guineensis) na Amazônia Oriental

A região amazônica tem sido submetida a contínuo desflorestamento e expansão do uso da terra, para a implantação de atividades como pecuária, exploração madeireira, agricultura, mineração e urbanização. A queima e a decomposição da biomassa da floresta liberam gases que contribuem para o efeito estufa, como o dióxido de carbono (CO₂). Nesse contexto, surge o interesse em avaliar a dinâmica do efluxo de CO₂ do solo na Amazônia, em especial em sistemas agroflorestais de palma de óleo (Elaeis guineensis), visto o destaque da produção dessa cultura no estado do Pará, que teve uma razoável expansão, transformando o estado do Pará em um dos maiores plantadores e produtores do país. Este trabalho teve como objetivo investigar a dinâmica do efluxo de CO₂ do solo em sistemas agroflorestais onde a palma de óleo é a cultura principal, na escala temporal e os fatores bióticos e abióticos que influenciam diretamente neste processo. Foram quantificados os carbono da biomassa microbiana do solo, carbono total do solo, respiração microbiana do solo, raízes finas do solo e zona de influência das espécies; e os fatores abióticos: umidade e temperatura do solo, em dois sistemas agroflorestais de cultivo de palma de óleo, o sistema adubadeiras e o sistema biodiverso. As medições foram feitas nos períodos seco e no chuvoso. Os resultados mostraram que o maior efluxo de CO₂ do solo ocorreu no período chuvoso, provavelmente devido à maior atividade microbiana nesse período influenciada por fatores climáticos aliados a fatores bióticos. O sistema biodiverso apresentou maior efluxo de CO₂ do solo do que o sistema adubadeiras, provavelmente devido à maior atividade biológica no solo nesse sistema. O efluxo de CO₂ do solo não mostrou correlação forte com as variáveis testadas. Pôde-se concluir que o efluxo de CO₂ do solo sofreu influencia apenas da sazonalidade climática. O fato da área de plantio ser muito jovem pode ser um fator determinante para que não tenha sido encontrada relação mais forte da respiração do solo com as variáveis analisadas.

Emissão de dióxido de carbono e estoque de carbono em sistemas convencionais e alternativo de produção agropecuária no Nordeste paraense

A aplicação de técnicas menos agressivas ao meio ambiente, como o uso de sistemas alternativos (corte e trituração), no lugar dos sistemas convencionais (corte-e-queima), além de favorecer o equilíbrio dos ciclos biogeoquímicos em áreas florestais, contribui para a mitigação das mudanças climáticas. O objetivo deste estudo foi estimar a emissão e o estoque de carbono do solo em sistemas de produção agropecuária em unidades rurais familiares do Nordeste paraense. Os estudos foram conduzidos em área de agricultor familiar no Município de Mãe do Rio, com temperatura média anual de 25 a 28°C, precipitação pluviométrica acimade 2500 mm e com solo predominante do tipo Latossolo Amarelo distrófico de textura média a argilosa. Foram selecionados 3 sistemas de uso da terra (cultivo com Schizolobium amazonicum, roça e silvipastoril) e mais uma área de referencia (floresta secundária), com 4 parcelas, medindo 20 m x 20 m cada. Foram avaliados a emissão de CO2 do solo, estoque de carbono no solo, estoque da liteira no solo e estoque de carbono na liteira. Os dados foram submetidos à Análise de Variância (ANOVA) e à comparação das médias pelo teste de Tukey, ao nível de 5%. Em todos os sistemas avaliados, as maiores emissões de CO2 do solo, observadas no período chuvoso, foram no sistema silvipastoril (5,02 Wmol CO2 m^-2 s^-1), em comparação à área da floresta secundária (3,56 Wmol CO2 m^-2 s^-1). De todas as áreas estudadas a maior emissão anual foi encontrada no sistema silvipastoril. O estoque de carbono no solo foi maior na área da floresta secundária, com total de 157± 31,10 Mg ha^-1 (0-100 cm). O maior estoque de liteira no solo encontrado foi para a fração da liteira não-lenhosa, em todos os sistemas agropecuários e floresta secundária. O maior estoque de carbono na liteira não-lenhosa total foi observada no mês de janeiro/2010, com média geral de 4,31± 3,0 Mg ha^-1, em todos os sistemas avaliados. Os sistemas de uso da terra que não utilizam o fogo no preparo da área, como os sistemas alternativos de corte-e-trituração, além de contribuirem para a mitigação das mudanças climáticas, ajudam na manutenção do funcionamento adequado dos ciclos biogeoquímicos nos ecossistemas terrestres.