Análises da expressão gênica por PCR quantitativo em tempo real (QPCR) de genes candidatos para a tolerância à seca em cafeeiro.

A seca é uma das principais limitações climáticas à produção do cafeeiro. A importância dessa limitação deve aumentar, em função das mudanças reconhecidas no clima global e, também, porque a cafeicultura vem sendo expandida para regiões marginais onde a baixa pluviosidade e temperaturas desfavoráveis se constituem grandes limitações à produção do café. A seca induz diversas respostas fisiológicas e moleculares nas plantas, incluindo alterações da expressão gênica, visando atingir ajuste osmótico, a indução de reparadores de sistemas moleculares e a expressão de diversas proteínas protetoras. Existem diversas formas de se avaliar a expressão gênica em plantas submetidas ao estresse hídrico. Uma técnica amplamente utilizada nos últimos anos, para a análise quantitativa da expressão gênica é a de PCR quantitativo em tempo real (qPCR). O objetivo do presente trabalho foi analisar a expressão de 14 genes candidatos para a tolerância à seca em cafeeiro, pré-selecionados em estudos de macroarranjos de cDNA, utilizando clones de Coffea canephora (Clone 22 ? sensível e clones 14, 73 e 120 ? tolerantes ao estresse hídrico)

Distribution of the Octopamine Receptor AmOA1 in the Honey Bee Brain

Octopamine plays an important role in many behaviors in invertebrates. It acts via binding to G protein coupled receptors located on the plasma membrane of responsive cells. Several distinct subtypes of octopamine receptors have been found in invertebrates, yet little is known about the expression pattern of these different receptor subtypes and how each subtype may contribute to different behaviors. One honey bee (Apis mellifera) octopamine receptor, AmOA1, was recently cloned and characterized. Here we continue to characterize the AmOA1 receptor by investigating its distribution in the honey bee brain. We used two independent antibodies produced against two distinct peptides in the carboxyl-terminus to study the distribution of the AmOA1 receptor in the honey bee brain. We found that both anti-AmOA1 antibodies revealed labeling of cell body clusters throughout the brain and within the following brain neuropils: the antennal lobes; the calyces, pedunculus, vertical (alpha, gamma) and medial (beta) lobes of the mushroom body; the optic lobes; the subesophageal ganglion; and the central complex. Double immunofluorescence staining using anti-GABA and anti-AmOA1 receptor antibodies revealed that a population of inhibitory GABAergic local interneurons in the antennal lobes express the AmOA1 receptor in the cell bodies, axons and their endings in the glomeruli. In the mushroom bodies, AmOA1 receptors are expressed in a subpopulation of inhibitory GABAergic feedback neurons that ends in the visual (outer half of basal ring and collar regions) and olfactory (lip and inner basal ring region) calyx neuropils, as well as in the collar and lip zones of the vertical and medial lobes. The data suggest that one effect of octopamine via AmOA1 in the antennal lobe and mushroom body is to modulate inhibitory neurons

Medroxyprogesterone Acetate Alters Mycobacterium Bovis BCG-Induced Cytokine Production in Peripheral Blood Mononuclear Cells of Contraceptive Users

Most individuals latently infected with Mycobacterium tuberculosis (M.tb) contain the infection by a balance of effector and regulatory immune responses. This balance can be influenced by steroid hormones such as glucocorticoids. The widely used contraceptive medroxyprogesterone acetate (MPA) possesses glucocorticoid activity. We investigated the effect of this hormone on immune responses to BCG in household contacts of active TB patients. Multiplex bead array analysis revealed that MPA demonstrated both glucocorticoid and progestogenic properties at saturating and pharmacological concentrations in peripheral blood mononuclear cells (PBMCs) and suppressed antigen specific cytokine production. Furthermore we showed that PBMCs from women using MPA produced significantly lower levels of IL-1α, IL-12p40, IL-10, IL-13 and G-CSF in response to BCG which corresponded with lower numbers of circulating monocytes observed in these women. Our research study is the first to show that MPA impacts on infections outside the genital tract due to a systemic effect on immune function. Therefore MPA use could alter susceptibility to TB, TB disease severity as well as change the efficacy of new BCG-based vaccines, especially prime-boost vaccine strategies which may be administered to adult or adolescent women in the future