966 resultados para Direct counting by microscopy
Resumo:
Esta tese se insere dentro da leitura bíblica latino-americana trazendo, nessa ótica, a riqueza e o desafio do que significa ler a Bíblia a partir de um contexto de opressão como é o nosso continente. Nos servimos do roteiro das ciências bíblicas e dos métodos exegéticos modernos. É uma pesquisa bibliográfica onde, eventualmente, consideramos o nível experiêncial, por exemplo quando trabalhamos o tema do derramamento do Espírito nos pentecostais chilenos. Esse capítulo, IV, segue o roteiro da hipótese central de nossa tese na medida que firmamos o princípio de que os pentecostais fazem parte do amplo e diverso grupo dos enfraquecidos como produto de um sistema excludente, baseado na exploração social. Numa situação de crise, o anúncio do derramamento do Espírito é um sinal de salvação, de perspectivas futuras, de conservação e prolongação da vida. Esta tese relê o tema do derramamento do Espírito, a partir de Jl 3,1-5. O tema é analisado num contexto social concreto, onde os enfraquecidos recebem o Espírito do Senhor. O eixo que nos permite fazer esta leitura encontra sua referencial nas pessoas setores sociais - mencionadas como beneficiadas diretas da ação do Espírito. São pessoas que representam setores diferentes; jovens, escravos e escravas, formam o grupo que sustenta e faz funcionar o sistema imperial persa grego - baseado na exploração e mão de obra barata. Eles, juntamente com os idosos que não produzem mais, são a base de sustentação da pirâmide social. Como demostraremos no decorrer da tese, nossa hipótese somente é possível se duas conjunturas se cruzam. Para isso, em num primeiro momento, localizamos e demostramos que o livro de Joel deve ser situado no contexto da literatura apocalíptica (nascimento). Em seguida, demonstramos que o livro de Joel, como produto literário final, deve ser localizado no contexto histórico do pós-exílio, isto é, no tempo dos impérios persa e grego. A confirmação destes dois aspectos desenvolvidos nos capítulos I e II, nos permitem, na análise literária, capítulo III, aprofundar, mediante a análise exegética, e confirmar a nossa hipótese central. Com estes três capítulos demostramos que o derramamento do Espírito do Senhor tem como destinatários preferenciais, senão exclusivos, os setores enfraquecidos pela política social, econômica e religiosa dos impérios persa e grego. Com estes três capítulos desenvolvidos, no quarto capítulo analisamos uma experiência concreta, de um setor majoritariamente pobre que tem se apropriado do texto de Jl 3,1-5, e encontrado nele uma alternativa social, política e religiosa para se manter fiel ao Senhor e por quase um século recria e revive a experiência do derramamento do Espírito. Os velhos, os jovens, os escravos e as escravas, formam o setor dos enfraquecidos. É o setor que nada espera, e que nada terá da parte dos impérios e que, como os pentecostais, pela sua situação de enfraquecimento, acredita que somente uma intervenção externa pode mudar o seu futuro, pode trazer de volta a esperança. Essa intervenção externa começa a ser possível pelo derramamento do Espírito e se concretiza na chegada do dia de Javé, que será grande e terrível, onde o sol e a lua se unem para indicar o monte de Sião e a cidade de Jerusalém como lugar de adoração, refúgio e salvação. Esta perspectiva da ação do Espírito é possível somente no período do pós-exílio. Nesse tempo o Espírito adquire uma dimensão ampla e inclusiva agindo sobre diversos setores sociais, independe de serem ou não judeus.(AU)
Resumo:
Asparaginyl-tRNA (Asn-tRNA) and glutaminyl-tRNA (Gln-tRNA) are essential components of protein synthesis. They can be formed by direct acylation by asparaginyl-tRNA synthetase (AsnRS) or glutaminyl-tRNA synthetase (GlnRS). The alternative route involves transamidation of incorrectly charged tRNA. Examination of the preliminary genomic sequence of the radiation-resistant bacterium Deinococcus radiodurans suggests the presence of both direct and indirect routes of Asn-tRNA and Gln-tRNA formation. Biochemical experiments demonstrate the presence of AsnRS and GlnRS, as well as glutamyl-tRNA synthetase (GluRS), a discriminating and a nondiscriminating aspartyl-tRNA synthetase (AspRS). Moreover, both Gln-tRNA and Asn-tRNA transamidation activities are present. Surprisingly, they are catalyzed by a single enzyme encoded by three ORFs orthologous to Bacillus subtilis gatCAB. However, the transamidation route to Gln-tRNA formation is idled by the inability of the discriminating D. radiodurans GluRS to produce the required mischarged Glu-tRNAGln substrate. The presence of apparently redundant complete routes to Asn-tRNA formation, combined with the absence from the D. radiodurans genome of genes encoding tRNA-independent asparagine synthetase and the lack of this enzyme in D. radiodurans extracts, suggests that the gatCAB genes may be responsible for biosynthesis of asparagine in this asparagine prototroph.
Resumo:
Interferon γ (IFN-γ) induces rapid tyrosine phosphorylation of the latent cytoplasmic transcription factor, Stat1, which then forms homodimers, translocates to the nucleus and participates in IFN-γ-induced transcription. However, little is known of the interactions between Stat1 and the general transcription machinery during transcriptional activation. We show here that Stat1 can directly interact with the CREB-binding protein (CBP)/p300 family of transcriptional coactivators. Specifically, two interaction regions were identified: the amino-terminal region of Stat1 interacts with the CREB-binding domain of CBP/p300 and the carboxyl-terminal region of Stat1 interacts with the domain of CBP/p300 that binds adenovirus E1A protein. Transfection experiments suggest a role for these interactions in IFN-γ-induced transcription. Because CBP/p300-binding is required for the adenovirus E1A protein to regulate transcription of many genes during viral replication and cellular transformation, it is possible that the anti-viral effect of IFN-γ is based at least in part on direct competition by nuclear Stat1 with E1A for CBP/p300 binding.
Resumo:
A comparison was made of the competence for neoplastic transformation in three different sublines of NIH 3T3 cells and multiple clonal derivatives of each. Over 90% of the neoplastic foci produced by an uncloned transformed (t-SA′) subline on a confluent background of nontransformed cells were of the dense, multilayered type, but about half of the t-SA′ clones produced only light foci in assays without background. This asymmetry apparently arose from the failure of the light focus formers to register on a background of nontransformed cells. Comparison was made of the capacity for confluence-mediated transformation between uncloned parental cultures and their clonal derivatives by using two nontransformed sublines, one of which was highly sensitive and the other relatively refractory to confluence-mediated transformation. Transformation was more frequent in the clones than in the uncloned parental cultures for both sublines. This was dramatically so in the refractory subline, where the uncloned culture showed no overt sign of transformation in serially repeated assays but increasing numbers of its clones exhibited progressive transformation. The reason for the greater susceptibility of the pure clones is apparently the suppression of transformation among the diverse membership that makes up the uncloned parental culture. Progressive selection toward increasing degrees of transformation in confluent cultures plays a major role in the development of dense focus formers, but direct induction by the constraint of confluence may contribute by heritably damaging cells. In view of our finding of increased susceptibility to transformation in clonal versus uncloned populations, expansion of some clones at the expense of others during the aging process would contribute to the marked increase of cancer with age.
Resumo:
The p70 S6 kinase is activated by insulin and mitogens through multisite phosphorylation of the enzyme. One set of activating phosphorylations occurs in a putative autoinhibitory domain in the noncatalytic carboxyl-terminal tail. Deletion of this tail yields a variant (p70 delta CT104) that nevertheless continues to be mitogen regulated. Coexpression with a recombinant constitutively active phosphatidylinositol (PI) 3-kinase (EC 2.7.1.137) gives substantial activation of both full-length p70 and p70 delta CT104 but not Rsk. Activation of p70 delta CT104 by PI 3-kinase and inhibition by wortmannin are each accompanied by parallel and selective changes in the phosphorylation of p70 Thr-252. A Thr or Ser at this site, in subdomain VIII of the catalytic domain just amino-terminal to the APE motif, is necessary for p70 40S kinase activity. The inactive ATP-binding site mutant K123M p70 delta CT104 undergoes phosphorylation of Thr-252 in situ but does not undergo direct phosphorylation by the active PI 3-kinase in vitro. PI 3-kinase provides a signal necessary for the mitogen activation of the p70 S6 kinase, which directs the site-specific phosphorylation of Thr-252 in the p70 catalytic domain, through a distinctive signal transduction pathway.
Resumo:
Os microRNAs (miRNAs) são pequenos RNAs não codificadores de proteínas presentes na maioria dos eucariotos. Esses RNAs regulam a expressão gênica em nível pós-transcricional através do silenciamento de mRNAs-alvo que possuem sítios complementares às suas sequências, atuando em praticamente todos os processos celulares. Embora a estrutura e função dos miRNAs estejam bem caracterizadas, aspectos relacionados à sua organização genômica, evolução e atuação em doenças são tópicos que apresentam enormes lacunas. Nesta tese, utilizamos abordagens computacionais para investigar estes temas em três trabalhos. No primeiro, processamos e integramos um vasto volume de dados publicamente disponíveis referentes aos miRNAs e genes codificadores de proteínas para cinco espécies de vertebrados. Com isso, construimos uma ferramenta web que permite a fácil inspeção da organização genômica dos miRNAs em regiões inter e intragênicas, o acesso a dados de expressão de miRNAs e de genes codificadores de proteínas (classificados em genes hospedeiros e não hospedeiros de miRNAs), além de outras informações pertinentes. Verificamos que a ferramenta tem sido amplamente utilizada pela comunidade científica e acreditamos que ela possa facilitar a geração de hipóteses associadas à regulação dos miRNAs, principalmente quando estão inseridos em genes hospedeiros. No segundo estudo, buscamos compreender como o contexto genômico e a origem evolutiva dos genes hospedeiros influenciam a expressão e evolução dos miRNAs humanos. Nossos achados mostraram que os miRNAs intragênicos surgem preferencialmente em genes antigos (origem anterior à divergência de vertebrados). Observamos que os miRNAs inseridos em genes antigos têm maior abrangência de expressão do que os inseridos em genes novos. Surpreendentemente, miRNAs jovens localizados em genes antigos são expressos em um maior número de tecidos do que os intergênicos de mesma idade, sugerindo uma vantagem adaptativa inicial que pode estar relacionada com o controle da expressão dos genes hospedeiros, e como consequência, expondo-os a contextos celulares e conjuntos de alvos diversos. Na evolução a longo prazo, vimos que genes antigos conferem maior restrição nos padrões de expressão (menor divergência de expressão) para miRNAs intragênicos, quando comparados aos intergênicos. Também mostramos possíveis associações funcionais relacionadas ao contexto genômico, tais como o enriquecimento da expressão de miRNAs intergênicos em testículo e dos intragênicos em tecidos neurais. Propomos que o contexto genômico e a idade dos genes hospedeiros são fatores-chave para a evolução e expressão dos miRNAs. Por fim, buscamos estabelecer associações entre a expressão diferencial de miRNAs e a quimioresistência em câncer colorretal utilizando linhagens celulares sensíveis e resistentes às drogas 5-Fluoruracil e Oxaliplatina. Dentre os miRNAs identificados, o miR-342 apresentou níveis elevados de expressão nas linhagens sensíveis à Oxaliplatina. Com base na análise dos alvos preditos, detectamos uma significativa associação de miR-342 com a apoptose. A superexpressão de miR-342 na linhagem resistente SW620 evidenciou alterações na expressão de genes da via apoptótica, notavelmente a diminuição da expressão do fator de crescimento PDGFB, um alvo predito possivelmente sujeito à regulação direta pelo miR-342.
Resumo:
Delaware sets the governance standards for most public companies. The ability to attract corporations could not be explained solely by the existence of a favorable statutory regime. Delaware was not invariably the first or the only state to implement management friendly provisions. Given the interpretive gaps in the statute and the critical importance of the common law in the governance process, courts played an outsized role in setting legal standards. The management friendly nature of the Delaware courts contributed significantly to the state’s attraction to public corporations. A current example of a management friendly trend in the case law had seen the recent decisions setting out the board’s authority to adopt bylaws under Section 109 of the Delaware General Corporation Law (DGCL), particularly those involving the shifting of fees in litigation against the corporation or its directors. The DGCL allows bylaws that address “the business of the corporation, the conduct of its affairs, and its rights or powers or the rights or powers of its stockholders, directors, officers or employees.” The broad parameters are, however, subject to limits. Bylaws cannot be inconsistent with the certificate of incorporation or “the law.” Law includes the common law. The Delaware courts have used the limitations imposed by “the law” to severely restrict the reach of shareholder inspired bylaws. The courts have not used the same principles to impose similar restraints on bylaws adopted by the board of directors. This can be seen with respect to bylaws that restrict or even eliminate the right of shareholders to bring actions against management and the corporation. In ATP Tour, Inc. v. Deutscher Tennis Bund the court approved a fee shifting bylaw that had littl relationship to the internal affairs of the corporation. The decision upheld the bylaw as facially valid.The decision ignored a number of obvious legal infirmities. Among other things, the decision did not adequately address the requirement in Section 109(b) that bylaws be consistent with “the law.” The decision obliquely acknowledged that the provisions would “by their nature, deter litigation” but otherwise made no effort to assess the impact of this deterrence on shareholders causes of action. The provision in fact had the practical effect of restricting, if not eliminating, litigation rights granted by the DGCL and the common law. Perhaps most significantly, however, the bylaws significantly limited common law rights of shareholders to bring actions against the corporation and the board. Given the high dismissal rates for these actions, fee shifting bylaws imposed a meaningful risk of liability on plaintiffs. Moreover, because judgments in derivative suits were paid to the corporation, shareholders serving as plaintiffs confronted the risk of liability without any offsetting direct benefit. By preventing suits in this area, the bylaw effectively insulated the behavior of boards from legal challenge. The ATP decision was poorly reasoned and overstepped acceptable boundaries. The management friendly decision threatened the preeminent role of Delaware in the development of corporate law. The decision raised the specter of federal intervention and the potential for meaningful competition from the states. Because the opinion examined the bylaw in the context of non-stock companies, the reasoning may remain applicable only to those entities and never make the leap to for-profit stock corporations. Nonetheless, the analysis reflects a management friendly approach that does not adequately take into account the impact of the provision on the rights of shareholders.
Resumo:
This paper intends to illustrate the respective roles and functions of the Court of Justice of the EU (CJEU) on the one hand, and the Maltese national courts on the other. It will then define the scope and role of the judicial cooperation between the CJEU and the national courts, highlighting the procedure relating to the preliminary rulings. The paper will then briefly describe the cases brought before the CJEU involving Malta, including those concerning requests for preliminary rulings originating from Malta, and the direct actions by the European Commission before the Court of Justice, as well as those before the General Court. After a description of the rationale behind the publication of the book Malta u l-Qorti tal-Ġustizzja tal-Unjoni Ewropea (Malta and the Court of Justice of the European Union), and following the conference in which it was presented, the main points that emerged from the conference will serve as a backdrop to some statistical analysis pertaining to the Maltese cases, as well as some reflections on the current situation of the judicial cooperation obtained after ten years. It will propose that, besides a mere statistical analysis of the raw figures that emerge, one must rather address his attention to the spirit of EU membership, and reflect on whether Malta’s legal system has actually absorbed and understood the full meaning of the EU membership, ten years after it took place.
Resumo:
This paper empirically analyses a dataset of more than 7,300 agricultural land sales transactions from 2001 and 2007 to identify the factors influencing agricultural land prices in Bavaria. We use a general spatial model, which combines a spatial lag and a spatial error model, and in addition account for endogeneity introduced by the spatially lagged dependent variable as well as other explanatory variables. Our findings confirm the strong influence of agricultural factors such as land productivity, of variables describing the regional land market structure, and of non-agricultural factors such as urban pressure on agricultural land prices. Moreover, the involvement of public authorities as a seller or buyer increases sales prices in Bavaria. We find a significant capitalisation of government support payments into agricultural land, where a decrease of direct payments by 1% would decrease land prices in 2007 and 2001 by 0.27% and 0.06%, respectively. In addition, we confirm strong spatial relationships in our dataset. Neglecting this leads to biased estimates, especially if aggregated data is used. We find that the price of a specific plot increases by 0.24% when sales prices in surrounding areas increase by 1%.
Resumo:
The quantity, type, and maturity of the organic matter of Quaternary and Tertiary sediments from the Japan Trench (DSDP Leg 56, Sites 434 and 436; and Leg 57, Sites 438, 439 and 440) were determined. The hydrocarbons in lipid extracts were analyzed by capillary- column gas chromatography and combined gas chromatography/ mass spectrometry. Kerogen concentrates were investigated by microscopy, and vitrinite-reflectance values were determined. Measured organic-carbon values were in the range of 0.13 to 1.00 per cent. Extract yields, however, were extremely low. Normalized to organic carbon, total extracts ranged from 4.1 to 15.7 mg/g Corg. Gas chromatography of non-aromatic hydrocarbons showed that all sediments, except one Oligocene sample, contained very immature, mainly terrigenous organic material. This was indicated by n-alkane maxima at C29 and C31 and high odd-carbon-number predominances. Unsaturated steroid hydrocarbons were found to be major cyclic compounds in lower- and middle-Miocene samples from the upper inner trench slope (Sites 438 and 439). Perylene was the dominating aromatic hydrocarbon in all but the Oligocene sample. Microscopy showed kerogens rich in terrigenous organic particles, with a major portion of recycled vitrinite. Nevertheless, almost all the liptinite particles appeared to be primary. This is a paradox, as the bulk of the samples were composed of hemipelagic mineral matter with a major siliceous biogenic (planktonic) component. A trend of reduced size and increased roundness can be seen for the vitrinite/ inertinite particles from west to east (from upper inner trench slope to outer trench slope). All sediments but one are relatively immature, with mean huminite-reflectance values (Ro)in the range of 0.30 to 0.45 per cent. The oldest and deepest sediment investigated, an Oligocene sandstone from Site 439, yielded a mean vitrinitereflectance value of 0.74 per cent and a mature n-alkane distribution. This sample may indicate a geothermal event in late Oligocene time. It failed to affect the overlying lower Miocene and may have been caused by an intrusion. Boulders of acidic igneous rocks in the Oligocene can be interpreted as witnesses of nearby volcanic activity accompanied by intrusions.
Resumo:
On August 25, 2011, Governor Pat Quinn signed House Bill 1488 into law, now Public Act 97-0558 (The Act). The Act specifically directs a Management Improvement Initiative Committee (The Committee) to implement recommendations outlined in the January 2011 report to the General Assembly as required under Public Act 96-1141. The Act directs the group, formed under the auspices of Public Act 96-1141, to continue their work based on categories of recommendations. Each recommendation area has the common goal of reviewing providers from redundant monitoring, auditing, and reporting requirements. Implementing the recommendations of the Act will result in efficiency in business process for our providers, reinvestment of dollars saved from inefficient or unrealized administrative costs, and ultimately foster a network of sustainable human services providers in Illinois while increasing the level of direct service by the State agencies, contracted providers, and communities, who are all facing current economic pressures in the fiscal crisis.