gamma-Radiation induces apoptosis via sarcoplasmatic reticulum in guinea pig ileum smooth muscle cells

We investigated the effects of gamma-radiation on cells isolated from the longitudinal smooth muscle layer of the guinea pig ileum, a relatively radioresistant tissue. Single doses (up to 50 Gy) reduced the amount of sarcoplasmatic reticulum and condensed the myofibrils, as shown by electron microscopy 3 days post-irradiation. After that, contractility of smooth muscle strips was reduced. Ca(2+) handling was altered after irradiation, as shown in fura-2 loaded cells, with elevated basal intracellular Ca(2+), reduced amount of intrareticular Ca(2+), and reduced capacitive Ca(2+) entry. Radiation also induced apoptosis, judged from flow cytometry of cells loaded with proprium iodide. Electron microscopy showed that radiation caused condensation of chromatin in dense masses around the nuclear envelope, the presence of apoptotic bodies, fragmentation of the nucleus, detachment of cells from their neighbors, and reductions in cell volume. Radiation also caused activation of caspase 12. Apoptosis was reduced by the administration of the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone methyl ester (Z-VAD-FIVIK) during the 3 day period after irradiation, and by the chelator of intracellular Ca(2+), 1,2-bis(o-aminophenoxy)ethane-N,N,N`,N`-tetraacetic acid (BAPTA), from 1 h before until 2 h after irradiation. BAPTA also reduced the effects of radiation on contractility, basal intracellular Ca(2+), amount of intrareticular Ca(2+), capacitative Ca(2+) entry, and apoptosis. In conclusion, the effects of gamma radiation on contractility, Ca(2+) handling, and apoptosis appear due to a toxic action of intracellular Ca(2+). Ca(2+)-induced damage to the sarcoplasmatic reticulum seems a key event in impaired Ca(2+) handling and apoptosis induced by gamma-radiation. (c) 2008 Elsevier B.V. All rights reserved.

Polymorphisms at the glutathione S-transferase GSTM1, GSTT1 and GSTP1 loci: risk of ovarian cancer by histological subtype

The phase II glutathione S-transferases (GSTs) GSTT1, GSTM1 and GSTP1 catalyse glutathione-mediated reduction of exogenous and endogenous electrophiles. These GSTs have broad and overlapping substrate specificities and it has been hypothesized that allelic variants associated with less effective detoxification of potential carcinogens may confer an increased susceptibility to cancer. To assess the role of GST gene variants in ovarian cancer development, we screened 285 epithelial ovarian cancer cases and 299 unaffected controls for the GSTT1 deletion (null) variant, the GSTM1 deletion (null) variant and the GSTP1 codon 104 A-->G Ile-->Val amino acid substitution variant, The frequencies of the GSTT1, GSTM1 and GSTP1 polymorphic variants did not vary with tumour behaviour (low malignant potential or invasive) or p53 immunohistochemical status. There was a suggestion that ovarian cancers of the endometrioid or clear cell histological subtype had a higher frequency of the GSTT1 and GSTM1 deletion genotype than other histological subgroups. The GSTT1, GSTM1 and GSTP1 genotype distributions did not differ significantly between unaffected controls and ovarian cancer cases (overall or invasive cancers only). However, the GSTM1 null genotype was associated with increased risk of endometrioid/clear cell invasive cancer [age-adjusted OR (95% CI) = 2.04 (1.01-4.09), P = 0.05], suggesting that deletion of GSTM1 may increase the risk of ovarian cancer of these histological subtypes specifically. This marginally significant finding will require verification by independent studies.

Isolation and identification of the toxic peptides from Lophyrotoma zonalis (Pergidae) sawfly larvae

The broad-leaved paper bark tree Melaleuca quinquenervia (Cav) (Myrtaceae) was introduced into Florida (USA) early in this century it has proliferated to such an extent that urgent measures are now required to control it. The sawfly Lophyrotoma zonalis (Pergidae) has been introduced as a possible biological control agent due to its ability to defoliate M. quinquenervia. Because toxic D-amino acid- containing peptides have been isolated from some sawfly species, L. zonalis larvae were processed using the previously reported method for the recovery of these compounds. The toxins lophyrotomin (as the free C-terminal acid) and a mixture of pergidin and Val(4)-pergidin were isolated at 0.36 and 0.43% yield of the dried larvae, respectively. Both compounds when dosed intraperitoneally to C57/B16 male mice were hepatotoxic with lowest lethal doses of 8 and 32 mg/kg, respectively. The pathology of the liver was different for each compound, with the lophyrotomin free acid causing a periportal haemorrhagic necrosis and the pergidin causing a periacinar coagulative necrosis. (C) 2001 Elsevier Science Ltd. All rights reserved.

Specificity of 17 beta-oestradiol and benzo[a] pyrene oxidation by polymorphic human cytochrome P4501B1 variants substituted at residues 48, 119 and 432

1. Eight human cytochrome P4501B1 (CYP1B1) allelic variants, namely Arg(48)Ala(119)Leu(432), Arg(48)Ala(119)Val(432), Gly(48)Ala(119)Leu(432), Gly(48)Ala(119)Val(432), Arg(48)Ser(119)Leu(432), Arg(48)Ser(119)Val(432), Gly(48)Ser(119)Leu(432) and Gly(48)Ser(119)Val(432) (all with Asn(453)), were expressed in Escherichia coli together with human NADPH-P450 reductase and their catalytic specificities towards oxidation of 17 beta -oestradiol and benzo[a]pyrene were determined. 2. All of the CYP1B1 variants expressed in bacterial membranes showed Fe2+. CO versus Fe2+ difference spectra with wavelength maxima at 446 nm and they reacted with antibodies raised against recombinant human CYP1B1 in immunoblots. The ratio of expression of the reductase to CYP1B1 in these eight preparations ranged from 0.2 to 0.5. 3. CYP1B1 Arg(48) variants tended to have higher activities for 17 beta -oestradiol 4-hydroxylation than Gly(48) variants, although there were no significant variations in 17 beta -oestradiol 2-hydroxylation activity in these eight CYP1B1 variants. Interestingly, ratios of formation of 17 beta -oestradiol 4-hydroxylation to 2-hydroxylation by these CYP1B1 variants were higher in all of the Val(432) forms than the corresponding Leu(432) forms. 4. In contrast, Leu(432) forms of CYP1B1 showed higher rates of oxidation of benzo[a]pyrene (to the 7, 8-dihydoxy-7,8-dihydrodiol in the presence of epoxide hydrolase) than did the Val(432) forms. 5. These results suggest that polymorphic human CYP1B1 variants may cause some altered catalytic specificity with 17 beta -oestradiol and benzo[a]pyrene and may influence susceptibilities of individuals towards endogenous and exogenous carcinogens.

beta-strand mimicking macrocyclic amino acids: Templates for protease inhibitors with antiviral activity

New amino acids are reported in which component macrocycles are constrained to mimic tripeptides locked in a beta-strand conformation. The novel amino acids involve macrocycles functionalized with both an N- and a C-terminus enabling addition of appendages at either end to modify receptor affinity, selectivity, or membrane permeability. We show that the cycles herein are effective templates within inhibitors of HIV-1 protease. Eleven compounds originating from such bifunctionalized cyclic templates are potent inhibitors of HIV-1 protease (Ki 0.3-50 nM; pH 6.5, I = 0.1 M). Unlike normal peptides comprising amino acids, five of these macrocycle-containing compounds are potent antiviral agents with sub-micromolar potencies (IC50 170-900 nM) against HIV-1 replication in human MT2 cells. The most active antiviral agents are the most lipophilic, with calculated values of LogD(6.5) greater than or equal to 4. All molecules have a conformationally constrained 17-membered macrocyclic ring that has been shown to structurally mimic a tripeptide segment (Xaa)-(Val/Ile)-(Phe/Tyr) of a peptide substrate in the extended conformation. The presence of two trans amide bonds and a para-substituted aromatic ring prevents intramolecular hydrogen bonds and fixes the macrocycle in the extended conformation. Similarly constrained macrocycles may be useful templates for the creation of inhibitors for the many other proteins and proteases that recognize peptide beta-strands.

Identification of residues and domains of Raf important for function in vivo and in vitro

Random mutagenesis and genetic screens for impaired Raf function in Caenorhabditis elegans were used to identify six loss-of-function alleles of lin-45 raf that result in a substitution of a single amino acid. The mutations were classified as weak, intermediate, and strong based on phenotypic severity. We engineered these mutations into the homologous residues of vertebrate Raf-1 and analyzed the mutant proteins for their underlying biochemical defects. Surprisingly, phenotype strength did not correlate with the catalytic activity of the mutant proteins. Amino acid substitutions Val-589 and Ser-619 severely compromised Raf kinase activity, yet these mutants displayed weak phenotypes in the genetic screen. Interestingly, this is because these mutant Raf proteins efficiently activate the MAPK (mitogen-activated protein kinase) cascade in living cells, a result that may inform the analysis of knockout mice. Equally intriguing was the observation that mutant proteins with non-functional Ras-binding domains, and thereby deficient in Ras-mediated membrane recruitment, displayed only intermediate strength phenotypes. This confirms that secondary mechanisms exist to couple Ras to Raf in vivo. The strongest phenotype in the genetic screens was displayed by a S508N mutation that again did not correlate with a significant loss of kinase activity or membrane recruitment by oncogenic Ras in biochemical assays. Ser-508 lies within the Raf-1 activation loop, and mutation of this residue in Raf-1 and the equivalent Ser-615 in B-Raf revealed that this residue regulates Raf binding to MEK. Further characterization revealed that in response to activation by epidermal growth factor, the Raf-S508N mutant protein displayed both reduced catalytic activity and aberrant activation kinetics: characteristics that may explain the C. elegans phenotype.

The highly rearranged mitochondrial genome of the plague thrips, Thrips imaginis (Insecta : thysanoptera): Convergence of two novel gene boundaries and an extraordinary arrangement of rRNA genes

To help understand the mechanisms of gene rearrangement in the mitochondrial (mt) genomes of hemipteroid insects, we sequenced the mt genome of the plague thrips, Thrips imaginis (Thysanoptera). This genome is circular, 15,407 by long, and has many unusual features, including (1) rRNA genes inverted and distant from one another, (2) an extra gene for tRNA-Ser, (3) a tRNA-Val lacking a D-arm, (4) two pseudo-tRNA genes, (5) duplicate control regions, and (6) translocations and/or inversions of 24 of the 37 genes. The mechanism of rRNA gene transcription in T. imaginis may be different from that of other arthropods since the two rRNA genes have inverted and are distant from one another. Further, the rRNA genes are not adjacent or even close to either of the two control regions. Tandem duplication and deletion is a plausible model for the evolution of duplicate control regions and for the gene translocations, but intramitochondrial recombination may account for the gene inversions in T. imaginis. All the 18 genes between control regions #1 and #2 have translocated and/or inverted, whereas only six of the 20 genes outside this region have translocated and/or inverted. Moreover, the extra tRNA gene and the two pseudo-tRNA genes are either in this region or immediately adjacent to one of the control regions. These observations suggest that tandem duplication and deletion may be facilitated by the duplicate control regions and may have occurred a number of times in the lineage leading to T. imaginis. T. imaginis shares two novel gene boundaries with a lepidopsocid species from another order of hemipteroid insects, the Psocoptera. The evidence available suggests that these shared gene boundaries evolved by convergence and thus are not informative for the interordinal phylogeny of hemipteroid insects. We discuss the potential of hemipteroid insects as a model system for studies of the evolution of animal rut genomes and outline some fundamental questions that may be addressed with this system.

Diversidade e capacita????o em escolas de governo: mesa-redonda de pesquisa-a????o

A MRPA Diversidade e Capacita????o em Escolas de Governo reuniu especialistas e dirigentes de variadas origens para pensar a constru????o de alternativas para a inser????o dos temas diversidade, igualdade de g??nero, igualdade racial e direitos humanos nos programas de capacita????o e forma????o desenvolvidos em escolas de governo

Planejamento estrat??gico: contribui????o para o desempenho do Tribunal Superior do Trabalho

O presente estudo investiga em que medida o planejamento estrat??gico do Tribunal Superior do Trabalho contribuiu para o desempenho jurisdicional da Corte, em particular, no tocante ao n??mero de processos solucionados e ao prazo m??dio de tramita????o do processo na Corte. Para tanto, a pesquisa obedeceu ?? metodologia quantitativa, mediante a ado????o da t??cnica n??o param??trica da An??lise Envolt??ria de Dados ??? DEA. Tr??s cen??rios foram analisados, definidos de acordo com os inputs e outpts previamente eleitos. No cen??rio I, no per??odo de 2007 a 2012, o Tribunal Superior do Trabalho atingiu o score de 100% (cem por cento) nos anos de 2007, 2008, 2009 e 2012, ou seja, esses foram os anos em que a Corte alcan??ou os melhores resultados, considerados os inputs utilizados no processo produtivo (fatores de produ????o). Ressalta-se o fato de que os anos 2007, 2008 e 2009 s??o anteriores ?? implanta????o do planejamento estrat??gico no Tribunal. Por sua vez, nos cen??rios II e III, o score de 100% (cem por cento) foi atingido pela Corte nos anos de 2008, 2009 e 2012. Os cen??rios investigados revelaram que os anos anteriores ?? ado????o do planejamento estrat??gico, ?? exce????o de 2012, foram os de maior efici??ncia da Corte, sobretudo os anos de 2008 e 2009, que apresentaram score de 100% nas tr??s conjunturas. Diante desse desfecho, se, por um lado, n??o foi poss??vel relacionar a implanta????o do planejamento estrat??gico com o aumento da efici??ncia do Tribunal Superior do Trabalho, de outra parte, n??o se pode concluir, peremptoriamente, que n??o influenciou no rendimento. Prova disso, foi o ano de 2012, que se mostrou entre os de maior efici??ncia do TST, nos tr??s cen??rios. A pesquisa, portanto, resultou inconclusiva, possivelmente em virtude do pouco tempo de implanta????o do planejamento estrat??gico na Corte, que n??o permitiu a realiza????o da an??lise considerando-se um lapso temporal mais dilatado

Rela????es de parceria entre poder p??blico e entes de coopera????o e colabora????o no Brasil

O primeiro volume da s??rie Inova????o na Gest??o P??blica trata dos v??nculos de coopera????o entre a administra????o p??blica e entidades civis sem fins lucrativos, apresentando texto elaborado a partir de pesquisa financiada pela AECID sobre a dissemina????o no Brasil dos institutos do Servi??o Social Aut??nomo (SSA), da Organiza????o Social (OS) e da Organiza????o da Sociedade Civil de Interesse P??blico (OSCIP), no ??mbito dos governos federal, estadual e municipal. Esta publica????o est?? organizada como segue: o Cap??tulo I cont??m uma an??lise geral das rela????es entre o Poder P??blico e o chamado Terceiro Setor, desde as origens hist??ricas da atividade assistencial, e seus impactos no modelo assistencial brasileiro. O cap??tulo discute, tamb??m, o conceito de Terceiro Setor e os atuais formatos de financiamento das entidades civis que atuam no campo da assist??ncia social. O Cap??tulo II, por sua vez, contempla os resultados do diagn??stico das formas de relacionamento da Administra????o P??blica com entes de coopera????o e colabora????o, segundo pesquisa realizada entre novembro de 2008 e fevereiro de 2009 no ??mbito da coopera????o entre a extinta Secretaria de Gest??o (SEGES) e a AECID. O Cap??tulo III trata dos principais aspectos do processo de avalia????o e fiscaliza????o dos contratos de gest??o firmados entre o Poder Executivo Federal e as OS, com ??nfase no processo de presta????o de contas dos resultados do contrato

Proposta de taxonomias para ??rg??os e entidades da Administra????o P??blica Federal e outros entes de coopera????o e colabora????o

O quarto volume da s??rie Inova????o na Gest??o P??blica apresenta dois estudos que analisam a taxonomia para ??rg??os e entidades da Administra????o P??blica Federal e outros entes de coopera????o e colabora????o de perspectivas diferentes. O primeiro estudo, realizado pelo consultor Luiz Arnaldo Pereira da Cunha Junior, analisa e faz sugest??es de aperfei??oamento nos modelos das organiza????es e dos instrumentos de implementa????o de pol??ticas p??blicas, com foco no Poder Executivo. Ressalte-se, assim, que esse estudo n??o reflete as diretrizes ou orienta????es de governo. Trata-se de importante material de pesquisa que poder??, talvez, inspirar melhorias no modelo de aparelho de Estado no Brasil como resposta aos desafios de uma gest??o p??blica democr??tica voltada para resultados. O segundo estudo foi realizado pela Secretaria de Gest??o P??blica, de autoria de Val??ria Alpino Bigonha Salgado, com a colabora????o de Valdomiro Jos?? de Almeida. Consiste em um modelo taxon??mico de an??lise das formas de exerc??cio das fun????es executivas do Estado na regula????o social e na economia, constru??do a partir das disposi????es legais e normativas atualmente vigentes no ??mbito do Poder Executivo Federal

Democracia, direito e gest??o p??blica: textos para discuss??o

O quinto volume da s??rie Inova????o na Gest??o P??blica apresenta uma colet??nea de oito textos produzidos no contexto do Projeto Ciclos de Debates Direito e Gest??o P??blica, lan??ado em 2009, por meio da ent??o Secretaria de Gest??o e da Consultoria Jur??dica do Minist??rio do Planejamento, Or??amento e Gest??o, em parceria com a Associa????o Nacional dos Procuradores da Rep??blica, apoiada pelo Minist??rio P??blico Federal. O projeto promove o debate entre profissionais das diversas ??reas do conhecimento e experi??ncia, do setor p??blico e da sociedade, dando oportunidade ao confronto de conceitos e entendimento acerca da democracia, do direito e da gest??o p??blica. Os oito textos que constam na publica????o abordam variados temas. O primeiro desenvolve uma tese geral sobre a fun????o das atividades de consultoria jur??dica desempenhadas pela Advocacia de Estado no ??mbito da administra????o p??blica, com destaque para os contextos brasileiro e italiano. O segundo demonstra o processo de apropria????o da Constitui????o operado pela comunidade jur??dica e, sobretudo, pelos ??rg??os da Justi??a. Enquanto o terceiro texto aborda a a????o executiva do Estado na ??rea social, por meio do estabelecimento de v??nculos de coopera????o entre a Administra????o P??blica Direta e Indireta e entidades civis sem fins lucrativos. O quarto texto, por sua vez, apresenta o controle da legalidade dos atos do Poder Executivo e a realiza????o dos servi??os p??blicos, com destaque para o papel da advocacia p??blica em face da judicializa????o das decis??es administrativas. O quinto consiste na apresenta????o das tend??ncias recentes e quest??es em aberto sobre a ocupa????o no setor p??blico brasileiro, J?? o sexto trata da coopera????o internacional no contexto da inova????o e melhoria da gest??o p??blica. O s??timo apresenta a atual estrat??gia brasileira de inova????o dos modelos e instrumentos de gest??o p??blica. E o ??ltimo texto, o oitavo, encerra com o retrato hist??rico da previd??ncia social do servidor p??blico, desde a promulga????o da Constitui????o Federal de 1988 at?? a cria????o da Funda????o de Previd??ncia Complementar do Servidor P??blico Federal ??? Funpresp

Participa????o social: textos para discuss??o

O sexto volume da s??rie Inova????o na Gest??o P??blica apresenta uma colet??nea de tr??s textos sobre a quest??o da participa????o social. O primeiro texto, produzido pelo professor Mark Evans, no ??mbito da coopera????o do Minist??rio do Planejamento Or??amento e Gest??o com a Uni??o Europeia, fornece conhecimentos, tanto acad??micos quanto pr??ticos, sobre as pr??ticas europeias de ponta relacionadas ?? participa????o social. Isso envolve a aplica????o de uma gama de ferramentas para identificar e compartilhar melhores pr??ticas, diagnosticas o que funciona em diferentes contextos sociais, compatibilizar deferentes m??todos de envolvimento com diferentes fins e identificar onde o envolvimento do cidad??o pode ser ??til em quatro pontos de decis??o no processo pol??tico (dire????o estrat??gica, formula????o de pol??ticas, execu????o de pol??ticas e aprendizagem pol??tica). Al??m disso, o estudo tamb??m oferece uma sele????o de estudos de caso de toada Europa, a cada ponto de decis??o. O segundo texto, elaborado por Elisabete Ferrarezi e Clarice G. Oliveira e apresentado no V Congresso Consad de Gest??o P??blica, ocorrido em Bras??lia em junho de 2012, problematiza a efetividade dos espa??os e mecanismos de participa????o social do Brasil do ponto de vista do processamento de pol??ticas p??blicas dentro do aparato burocr??tico a partir de conceitos derivados das teorias de complexidade, participa????o e burocracia com a inten????o de contribuir para o debate sobre o aperfei??oamento desse campo. O ??ltimo texto, desenvolvido por N??ria Cunill Grau em setembro de 2012, no ??mbito da coopera????o entre o Minist??rio do Planejamento, Or??amento e Gest??o, o Banco Mundial e o Programa das Na????es Unidas para o Desenvolvimento (PNUD), cont??m um levantamento, realizado em 2009, dos diferentes instrumentos que facilitam a participa????o e o controle social na Administra????o P??blica Federal do Brasil associados aos diferentes modelos organizacionais existentes

Programa interativo de sa??de ocupacional

Em seu campus laboratorial de Xer??m, Duque de Caxias/RJ, o INMETRO herdou, do antigo Instituto Nacional de Pesos e Medidas, um t??mido servi??o de assist??ncia m??dica ??? composto por um m??dico e uma assistente social ??? insuficiente para atender aos seus 1.200 servidores e respectivos dependentes. Assim foi criado o Programa Interativo de Sa??de Ocupacional do INMETRO, baseado na explora????o de parcerias, na educa????o para a sa??de e no interc??mbio com ag??ncias governamentais. O check up anual tem se mostrado um dos itens mais importantes do programa, fundamental para o desenvolvimento de uma abordagem preventiva da sa??de. Os servidores t??m manifestado elevado grau de satisfa????o com a melhoria substancial da sua qualidade de vida, refletindo-se na qualidade dos servi??os da institui????o. Os servidores t??m-se envolvido intensamente nos eventos educativos promovidos no ??mbito do programa, como a Semana da Sa??de, com impacto sobre a comunidade em geral

Disciplina 4.7: monitoramento e avalia????o de pol??ticas p??blicas

Monitoramento e avalia????o de pol??ticas p??blicas: conceitua????o e tipos. O M&A em experi??ncias internacionais selecionadas. Formula????o, monitoramento e avalia????o de pol??ticas p??blicas no Brasil em contexto hist??rico. Experi??ncias de M&A de agendas estrat??gicas no Brasil no per??odo recente