Distribution of C-14 Cylindrospermopsin in vivo in the mouse

Radiolabelled C-14 cylindrospermopsin (CYN) has been prepared and used to investigate the distribution and excretion of CYN in vivo in male Quackenbush mice. At a dose of 0.2 mg/kg (i.e., approx. median lethal dose) the following mean (SID) urinary and faecal recoveries (cumulative) were obtained, respectively: (0-6 hours, n = 4) 48.2 (29.3)%, 11.9 (21.4)%; (0-12 hours, n = 12) 66.0 (27.1)%, 5.7 (5.6)%; (0-24 hours, n = 12) 68.4 (26.7)%, 8.5 (8.1)%. Mean (SD) recoveries from livers at 6 hours were 20.6 (6.4)% (n = 4), at 48 hours 13.1 (7.7)% (n = 8), and 5-7 days were 2.1 (2.1)% (n = 8). A substantial amount (up to 23%) can be retained in the liver for up to 48 hours with a lesser amount retained in the kidneys. The excretion patterns show substantial interindividual variability between predominantly faecal or urinary excretion, but these patterns are not related in any simple manner to the outcome in terms of toxicity. There is at least one methanol-extractable metabolite as well as a nonmethanol-extractable metabolite in the liver. The methanol-extractable metabolite was not found in the kidney and is more hydrophilic than CYN itself on reverse phase. (C) 2001 by John Wiley & Sons, Inc.

Nonspecific down-regulation of CD8+ T-Cell responses in mice expressing human Papillomavirus Type 16 E7 oncoprotein from the keratin-14 promoter

The E7 oncoprotein of human papillomavirus 16 (HPV16) transforms basal and suprabasal cervical epithelial cells and is a tumor-specific antigen in cervical carcinoma, to which immunotherapeutic strategies aimed at cytotoxic T-lymphocyte (CTL) induction are currently directed. By quantifying major histocompatibility complex class I tetramer-binding T cells and CTL in mice expressing an HPV16 E7 transgene from the keratin-l l (K14) promoter in basal and suprabasal keratinocytes and in thymic cortical epithelium, we show that antigen responsiveness of both E7- and non-E7-specific CD8(+) cells is down-regulation compared to non-E7 transgenic control mice. We show that the effect is specific for E7, and not another transgene, expressed from the K14 promoter, Down-regulation did not involve deletion of CD8(+) T cells of high affinity or high avidity, and T-cell receptor (TCR) VP-chain usage and TCR receptor density were similar in antigen-responsive cells from E7 transgenic and non-E7 transgenic mice. These data indicate that E7 expressed chronically from the K14 promoter nonspecifically down-regulates CD8+ T-cell responses. The in vitro data correlated with the failure of immunized E7 transgenic mice to control the growth of an E7-expressing tumor challenge, We have previously shown that E7-directed CTL down-regulation correlates with E7 expression in peripheral but not thymic epithelium (T, Dean et al., J, Virol. 73:6166-6170, 1999), The findings have implications for the immunological consequences of E7-expressing tumor development and E7-directed immunization strategies. Generically, the findings illustrate a T-cell immunomodulatory function for a virally encoded human oncoprotein.

Proteomic analysis reveals that 14-3-3 sigma in downregulated in human breast cancer cells

The class of molecular chaperones known as 14-3-3 is involved in the control of cellular growth by virtue of its apparent regulation of various signaling pathways, including the Raf/mitogen-activated protein kinase pathway. In breast cancer cells, the sigma form of 14-3-3 has been shown to interact with cyclin-dependent kinases and to control the rate of entry into mitosis. To test for a direct role for 14-3-3 in breast epithelial cell neoplasia, me have quantitated 14-3-3 protein levels using a proteomic approach based on two-dimensional electrophoresis and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF). We show here that 14-3-3 sigma protein is strongly down-regulated in the prototypic breast cancer cell lines MCF-7 and MDA-MB-231 and in primary breast carcinomas as compared with normal breast epithelial cells. In contrast, levels of the alpha, beta, delta, or zeta isoforms of 14-3-3 mere the same in both normal and transformed cells. The data support the idea that 14-3-3 sigma is involved in the neoplastic transition of breast epithelial cells by virtue of its role as a tumor suppressor; as such, it may constitute a robust marker with clinical efficacy for this pathology.

A novel 14-kilodalton protein interacts with the mitogen-activated protein kinase scaffold MP1 on a late endosomal/lysosomal compartment

We have identified a novel, highly conserved protein of 14 kD copurifying with late endosomes/lysosomes on density gradients. The protein, now termed p14, is peripherally associated with the cytoplasmic face of late endosomes/lysosomes in a variety of different cell types. In a two-hybrid screen with p14 as a bait, we identified the mitogen-activated protein kinase (MAPK) scaffolding protein MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK) partner 1 (MP1) as an interacting protein. We confirmed the specificity of this interaction in vitro by glutathione S-transferase pull-down assays and by coimmunoprecipitation, cosedimentation on glycerol gradients, and colocalization. Moreover, expression of a plasma membrane-targeted p14 causes mislocalization of coexpressed MP1. In addition, we could reconstitute protein complexes containing the p14-MP1 complex associated with ERK and MEK in vitro. The interaction between p14 and MP1 suggests a MAPK scaffolding activity localized to the cytoplasmic surface of late endosomes/lysosomes, thereby combining catalytic scaffolding and subcellular compartmentalization as means to modulate MAPK signaling within a cell.

Kainic acid induces 14-3-3 zeta expression in distinct regions of rat brain

Areas of the limbic system of adult male Wistar rats were screened for kainic-acid-induced gene expression. Polymerase-chain-reactionbased differential display identified a 147-bp cDNA fragment, which represented an mRNA that was upregulated in the entorhinal cortex and hippocampus in the kainic-acid-treated animals. The sequence was 97.8% homologous to rat 14-3-3 zeta isoform mRNA. Detailed Northern analysis revealed increased mRNA levels in the entorhinal cortex I h after kainic acid exposure and continued elevation 24 h post-injection in both the entorhinal cortex and hippocampus. Western blot analyses confirmed that the protein product of this gene was also present in increased amounts over the same time period. Immunohistochemistry and terminal transferase-mediated dUTP nick end labelling (TUNEL) detected expression of 14-3-3 protein exclusively in the entorhinal cortex and hippocampus, and only in TUNEL-positive neuronal cells. Expression of the tumor suppressor protein, p53 was also induced by kainate injection, and was co-localized with 14-3-3 zeta protein in selected cells only in the affected brain regions. The increase gene expression of 14-3-3 represents a transcription-mediated response associated with region selective neuronal damage induced by kainic acid. (C) 2002 Elsevier Science B.V. All rights reserved.

Structure of CcmG/DsbE at 1.14 angstrom resolution: High-fidelity reducing activity in an indiscriminately oxidizing environment

CcmG is unlike other periplasmic thioredoxin (TRX)like proteins in that it has a specific reducing activity in an oxidizing environment and a high fidelity of interaction. These two unusual properties are required for its role in c-type cytochrome maturation. The crystal structure of CcmG reveals a modified TRX fold with an unusually acidic active site and a groove formed from two inserts in the fold. Deletion of one of the groove-forming inserts disrupts c-type cytochrome formation. Two unique structural features of CcmG-an acidic active site and an adjacent groove-appear to be necessary to convert an indiscriminately binding scaffold, the TRX fold, into a highly specific redox protein.

Nitrate in soil humic acids revealed by N-14 nuclear magnetic resonance spectroscopy

Ecosystem management such as plant residue retention and prescribed burning can significantly affect soil organic matter (SOM) composition and, thereby, the closely associated carbon (C) and nitrogen (N) cycling processes, which underpin terrestrial ecosystem productivity and sustainability. Humic acid (HA) is an important SOM component and its chemical composition has attracted much attention. Here we report the first application of N-14 nuclear magnetic resonance (NMR) spectroscopy to soil HA study, revealing the surprising existence of nitrate-N and ammonia-N in the HAs. This newly discovered HA nitrate-N, though in a relatively low concentrations, is closely related to soil N availability and responsive to plant residue management regimes in contrasting forest ecosystems. The HA nitrate-N may be a useful and sensitive biochemical indicator of SOM quality in response to different ecosystem management regimes.

14-3-3 acts as an intramolecular bridge to regulate cdc25B localization and activity

One of the major regulators of mitosis in somatic cells is cdc25B. cdc25B is tightly regulated at multiple levels. The final activation step involves the regulated binding of 14-3-3 proteins. Previous studies have demonstrated that Ser-323 is a primary 14-3-3 binding site in cdc25B, which influences its activity and cellular localization. 14-3-3 binding to this site appeared to interact with the N-terminal domain of cdc25B to regulate its activity. The presence of consensus 14-3-3 binding sites in the N-terminal domain suggested that the interaction is through direct binding of the 14-3-3 dimer to sites in the N-terminal domain. We have identified Ser-151 and Ser-230 in the N-terminal domain as functional 14-3-3 binding sites utilized by cdc25B in vivo. These low affinity sites cooperate to bind the 14-3-3 dimer bound to the high affinity Ser-323 site, thus forming an intramolecular bridge that constrains cdc25B structure to prevent access of the catalytic site. Loss of 14-3-3 binding to either N-terminal site relaxes cdc25B structure sufficiently to permit access to the catalytic site, and the nuclear export sequence located in the N-terminal domain. Mutation of the Ser-323 site was functionally equivalent to the mutation of all three sites, resulting in the complete loss of 14-3-3 binding, increased access of the catalytic site, and access to nuclear localization sequence.

A????es premiadas no 14?? Concurso Inova????o na Gest??o P??blica Federal 2009

Nesta publica????o, referente ao 14?? Concurso Inova????o na Gest??o P??blica Federal, o leitor encontrar?? iniciativas inovadoras nas seguintes ??reas: Arranjos institucionais para coordena????o e/ou implementa????o de pol??ticas p??blicas (intra e inter-governamental); avalia????o e monitoramento de pol??ticas p??blicas; gest??o da informa????o; gest??o e desenvolvimento de pessoas; melhoria de processos de trabalho; planejamento, gest??o e desempenho institucional

Texto para discuss??o 14: gest??o e avalia????o de pol??ticas sociais: subs??dios para discuss??o

O texto procura refletir sobre algumas quest??es ligadas ??s problem??ticas de gest??o e avalia????o de pol??ticas sociais, a partir de conhecimentos relacionados com a observa????o da realidade brasileira. Trata-se de um alinhavado de considera????es divididas em quatro m??dulos: contextualiza????o, balan??o institucional, gargalos e novas abordagens. No primeiro ?? realizado um breve apanhado do contexto, em termos da administra????o p??blica, no qual se encontra situada a problem??tica social e o significado desta inser????o. O segundo traz um breve coment??rio sobre o endere??amento institucional das pol??ticas sociais e um balan??o de alguns de seus impasses. Em seguida, s??o indicados determinados gargalos que constituem estrangulamentos respons??veis pela incapacita????o estrutural do Estado no enfrentamento de certos problemas. S??o, ent??o, exploradas novas abordagens potenciais de enfrentamento e supera????o destas dificuldades. Finalmente ?? apresentado um conjunto de conclus??es, procurando estabelecer poss??veis conex??es com os esfor??os, no sentido de se ampliar a efici??ncia, a efic??cia e a efetividade das pol??ticas sociais atrav??s da descentraliza????o

M??dulo 14 - o sistema de registro de pre??os: curso legisla????o aplicada ?? log??stica de suprimentos Lei 8.666/93, preg??o e registro de pre??os

Este documento trata de: aspectos conceituais da lei: finalidade, import??ncia e hierarquia da lei; no????es gerais da lei de licita????es - Lei n?? 8.666/93; tipos de licita????o:menor pre??o, melhor t??cnica, t??cnica e pre??o e maior lance ou oferta; modalidades de licita????o: concorr??ncia, tomada de pre??os, convite, concurso e leil??o; exce????es ?? obrigatoriedade de licita????o: dispensa e inexigibilidade; regime de execu????o indireta; comiss??o de licita????o; etapas do processo licitat??rio: edital, procedimentos/documentos do certame, registro cadastral, habilita????o dos interessados, julgamento e encerramento; preg??o; registro de pre??os

O significado da maternidade para mães de 10 a 14 anos

A maternidade em meninas de 10 a 14 anos é um fenômeno que desafia as políticas públicas de saúde e a academia científica, visto que, sua ocorrência aumenta a cada ano, mesmo com a taxa de fecundidade decrescendo cada vez mais no Brasil. Procurando se aproximar dessa temática para contribuir com o aprofundamento do assunto nessa faixa etária, o presente estudo teve como objetivo compreender o significado da maternidade para mães de 10 a 14 anos de idade a partir de seus relatos de experiência. Parte-se de uma investigação epidemiológica realizada em uma região interiorana do Espírito Santo, no período de Julho de 2012 a Fevereiro de 2013, em que se avaliou a assistência pré-natal de 742 puérperas, através de informações registradas em formulário e contidas no Cartão da Gestante e no prontuário do recém-nascido. Aprofunda-se o estudo dos dez casos relativos às mães que possuíam entre 10 e 14 anos no momento do parto, mediante uma abordagem qualitativa que utilizou entrevistas individuais com roteiro semiestruturado. As falas foram gravadas, transcritas e analisadas através da técnica de análise de conteúdo, com enfoque na análise temática. Os resultados evidenciaram dois temas e cada um deles definiu quatro categorias de análise: 1) A experiência da maternidade: a) A vida familiar e conjugal; b) O que é ser mãe?; c) O apoio recebido após a maternidade e d) Reflexões sobre ser mãe adolescente e 2) A perspectiva após a maternidade: a) A escolaridade; b) A inserção no mercado de trabalho; c) Conselhos para jovens da mesma faixa etária e d) Os planos para o futuro. O estudo considera que o comportamento da maternidade poderá ter características e significados distintos de acordo com o contexto social em que vivem essas meninas. O arcabouço sexual, reprodutivo e familiar possui faces e formas estruturais que diferem dentro de cada grupo e a raiz desse assunto pode estar sendo baseada em uma relação que não depende da idade cronológica das mães.