Regulation by aldosterone of Na+,K+-ATPase mRNAs, protein synthesis, and sodium transport in cultured kidney cells.

Transepithelial Na+ reabsorption across tight epithelia is regulated by aldosterone. Mineralocorticoids modulate the expression of a number of proteins. Na+,K+-ATPase has been identified as an aldosterone-induced protein (Geering, K., M. Girardet, C. Bron, J. P. Kraehenbuhl, and B. C. Rossier, 1982, J. Biol. Chem., 257:10338-10343). Using A6 cells (kidney of Xenopus laevis) grown on filters we demonstrated by Northern blot analysis that the induction of Na+,K+-ATPase was mainly mediated by a two- to fourfold accumulation of both alpha- and beta-subunit mRNAs. The specific competitor spironolactone decreased basal Na+ transport, Na+,K+-ATPase mRNA, and the relative rate of protein biosynthesis, and it blocked the response to aldosterone. Cycloheximide inhibited the aldosterone-dependent sodium transport but did not significantly affect the cytoplasmic accumulation of Na+,K+-ATPase mRNA induced by aldosterone.

Genetic polymorphisms of the main transcription factors for adiponectin gene promoter in regulation of adiponectin levels: association analysis in three European cohorts.

Adiponectin serum concentrations are an important biomarker in cardiovascular epidemiology with heritability etimates of 30-70%. However, known genetic variants in the adiponectin gene locus (ADIPOQ) account for only 2%-8% of its variance. As transcription factors are thought to play an under-acknowledged role in carrying functional variants, we hypothesized that genetic polymorphisms in genes coding for the main transcription factors for the ADIPOQ promoter influence adiponectin levels. Single nucleotide polymorphisms (SNPs) at these genes were selected based on the haplotype block structure and previously published evidence to be associated with adiponectin levels. We performed association analyses of the 24 selected SNPs at forkhead box O1 (FOXO1), sterol-regulatory-element-binding transcription factor 1 (SREBF1), sirtuin 1 (SIRT1), peroxisome-proliferator-activated receptor gamma (PPARG) and transcription factor activating enhancer binding protein 2 beta (TFAP2B) gene loci with adiponectin levels in three different European cohorts: SAPHIR (n = 1742), KORA F3 (n = 1636) and CoLaus (n = 5355). In each study population, the association of SNPs with adiponectin levels on log-scale was tested using linear regression adjusted for age, sex and body mass index, applying both an additive and a recessive genetic model. A pooled effect size was obtained by meta-analysis assuming a fixed effects model. We applied a significance threshold of 0.0033 accounting for the multiple testing situation. A significant association was only found for variants within SREBF1 applying an additive genetic model (smallest p-value for rs1889018 on log(adiponectin) = 0.002, β on original scale = -0.217 µg/ml), explaining ∼0.4% of variation of adiponectin levels. Recessive genetic models or haplotype analyses of the FOXO1, SREBF1, SIRT1, TFAPB2B genes or sex-stratified analyses did not reveal additional information on the regulation of adiponectin levels. The role of genetic variations at the SREBF1 gene in regulating adiponectin needs further investigation by functional studies.

Trigo EMBRAPA 41: Nova cultivar para Minas Gerais, Goiás e Distrito Federal

A cultivar de trigo (Triticum aestivum L.) EMBRAPA 41 foi criada pela Embrapa-Centro de Pesquisa Agropecuária dos Cerrados (CPAC), em Planaltina, DF. Essa cultivar é o resultado da seleção realizada na descendência do cruzamento entre PF 813 e Polo 1; foi avaliada na fase experimental como linhagem CPAC 88118. A nova cultivar foi recomendada, pela Comissão Centro-Brasileira de Pesquisa de Trigo, para cultivo irrigado durante a estação seca, em 1995, nos estados de Minas Gerais e Goiás e no Distrito Federal. Essa cultivar destaca-se por apresentar estatura média, ciclo precoce, alto potencial de rendimento e por sua superior qualidade industrial. Nas avaliações de resistência a enfermidades, apresentou reação de resistência em relação à ferrugem-do-colmo, causada por Puccinia graminis tritici.

Aspectos fenológicos de seis espécies vegetais em matas de galeria do Distrito Federal

A fenologia de Vochysia piramidalis, Tapirira guianensis, Talauma ovata, Tococa formicaria, Miconia pseudonervosa e Miconia chamissois foi acompanhada de abril de 1994 a setembro de 1995, em dez indivíduos de cada espécie. Todas foram perenifólias, sugerindo relação com a presença de água no solo. Vochysia piramidalis e T. guianensis apresentaram padrão de floração atrasada em relação ao início das chuvas; T. ovata, T. formicaria e M. pseudonervosa, floração precoce; e M. chamissois, floração tardia. Quanto à duração da floração, M. pseudonervosa e T. formicaria apresentaram período longo e V. piramidalis, T. ovata, T. guianensis e M. chamissois, período curto. Vochysia piramidalis revelou padrão de dispersão anemocórico, com curto período de formação e maturação dos frutos. As demais, padrão zoocórico. Miconia pseudonervosa apresentou maturação variando dentro e entre indivíduos. Talauma ovata exibiu frutos imaturos por período extenso, mas com maturação rápida e simultânea no mesmo indivíduo. A produção foi abundante em T. guianensis, com maturação simultânea dentro do mesmo indivíduo e intervalos curtos entre os períodos reprodutivos. T. formicaria e M. chamissois apresentaram um período relativamente curto de frutificação, porém com maturação não-simultânea dentro do mesmo indivíduo. A frutificação das espécies zoocóricas foi durante o período chuvoso, coincidindo com o período de dispersão de animais.

Comparative study of the LC-MS/MS and UPLC-MS/MS for the multi-residue analysis of quinolones, penicillins and cephalosporins in cow milk, and validation according to the regulation 2002/657/EC

The aim of this study was to develop and validate an analytical method to simultaneously determine European Union-regulated beta-lactams (penicillins and cephalosporins) and quinolones in cow milk. The procedure involves a new solid phase extraction (SPE) to clean-up and pre-concentrate the three series of antibiotics before analysis by liquid chromatography¿tandem mass spectrometry (LC-MS/MS) and ultra-high-performance liquid chromatography¿tandem mass spectrometry (UPLC-MS/MS). LC-MS/MS and UPLC-MS/MS techniques were also compared. The method was validated according to the Directive 2002/657/EC and subsequently applied to 56 samples of raw cow milk supplied by the Laboratori Interprofessional Lleter de Catalunya (ALLIC) (Laboratori Interprofessional Lleter de Catalunya, Control Laboratory Interprofessional of Milk of Catalunya).

Regulation of peptidase activity in a three-dimensional aggregate model of brain tumor vasculature.

The abnormal vascular system of brain cancers inappropriately expresses membrane proteins, including proteolytic enzymes, ultimately resulting in blood extravasation. The production of inflammatory mediators, such as cytokines and nitric oxide, and tumor hypoxia have been implicated in these effects. We have previously shown that the activity of aminopeptidase A is increased in the abnormal vascular system of human and rat brain tumors. To study the mechanisms regulating the activities of peptidases in cerebral vasculature in brain tumors, we have developed a three-dimensional model of differentiated rat brain cells in aggregate cultures in which rat brain microvessels were incorporated. The secretion of interleukin-6 (IL-6) in the culture medium of aggregates was used as an indicator of inflammatory activation. Addition to these aggregates of C6 glioma cell medium (C6-CM) conditioned under hypoxic or normoxic conditions or serum mimicked tumor-dependent hypoxia or conditions of dysfunction of brain tumor vasculature. Hypoxic and normoxic C6-CM, but not serum, regulated peptidase activity in aggregates, and in particular it increased the activity of aminopeptidase A determined using histoenzymography. Serum, but not C6-CM, increased IL-6 production, but did not increase aminopeptidase A activity in aggregates. Thus soluble glioma-derived factors, but not serum-derived factors, induce dysfunctions of cerebral vasculature by directly regulating the activity of peptidases, not involving inflammatory activation. Tumor hypoxia is not necessary to modulate peptidase activity.

Adsorção de nitrato em solos de cerrado do Distrito Federal

Os objetivos deste trabalho foram verificar a adsorção de nitrato e identificar as propriedades do solo que mais influenciam este fenômeno, em um Latossolo Vermelho-Escuro, com 56% de argila, em Planaltina, DF. Em 1996 foram coletadas amostras de solo nas profundidades de 0-20, 20-40, 40-80, 80-100 cm, de um experimento com cinco anos de duração, em uma área de cerrado típico e em duas áreas cultivadas com soja e milho, onde o solo apresentava inversão de carga em profundidade. Também foram coletadas amostras em um solo sob cerradão, com carga elétrica líquida negativa em profundidade. A relação entre nitrato adsorvido e nitrato na solução foi descrita pela equação de Freundlich e as relações entre o nitrato adsorvido e algumas propriedades físico-químicas do solo foram estudadas por meio de regressões múltiplas. O nitrato adsorvido aumentou com a profundidade em todos os tratamentos. Esta adsorção foi maior nos solos sob cerrado e cerradão. O deltapH não afetou a adsorção de nitrato. A correlação entre nitrato adsorvido, matéria orgânica e sulfato extraível foi negativa. A adsorção de nitrato aumentou com a redução da matéria orgânica em profundidade e com a redução de sulfato extraível.

Differential regulation of Na-K-ATPase isoform gene expression by T3 during rat brain development.

A fetal rat telencephalon organotypic cell culture system was found to reproduce the developmental pattern of Na-K-adenosinetriphosphatase (ATPase) gene expression observed in vivo [Am. J. Physiol. 258 (Cell Physiol. 27): C1062-C1069, 1990]. We have used this culture system to study the effects of triiodothyronine (T3; 0.003-30 nM) on mRNA abundance and basal transcription rates of Na-K-ATPase isoforms. Steady-state mRNA levels were low at culture day 6 (corresponding to the day of birth) but distinct for each isoform alpha 3 much greater than beta 1 = beta 2 greater than alpha 2 greater than alpha 1. At culture day 6, T3 did not modify mRNA abundance of any isoform. At culture day 12 (corresponding to day 7 postnatal), T3 increased the mRNA level of alpha 2 (4- to 7-fold), beta 2 (4- to 5-fold), alpha 1 (3- to 6-fold), and beta 1 (1.5-fold), whereas alpha 3 mRNA levels remained unchanged. Interestingly, the basal transcription rate for each isoform differed strikingly (alpha 2 greater than alpha 1 much greater than beta 1 = beta 2 greater than alpha 3) but remained stable throughout 12 days of culture and was not regulated by T3. Thus we observed an inverse relationship between rate of transcription and rate of mRNA accumulation for each alpha-isoform, suggesting that alpha 1- and alpha 2-mRNA are turning over rapidly whereas alpha 3-mRNA is turning over slowly. Our data indicate that one of the mechanisms by which T3 selectively controls Na-K-ATPase gene expression during brain development in vitro occurs at the posttranscriptional level.

Epigenetic regulation of the bacterial cell cycle.

N(6)-methyl-adenines can serve as epigenetic signals for interactions between regulatory DNA sequences and regulatory proteins that control cellular functions, such as the initiation of chromosome replication or the expression of specific genes. Several of these genes encode master regulators of the bacterial cell cycle. DNA adenine methylation is mediated by Dam in gamma-proteobacteria and by CcrM in alpha-proteobacteria. A major difference between them is that CcrM is cell cycle regulated, while Dam is active throughout the cell cycle. In alpha-proteobacteria, GANTC sites can remain hemi-methylated for a significant period of the cell cycle, depending on their location on the chromosome. In gamma-proteobacteria, most GATC sites are only transiently hemi-methylated, except regulatory GATC sites that are protected from Dam methylation by specific DNA-binding proteins.