Increased aerobic metabolism is essential for the beneficial effects of caloric restriction on yeast life span

Calorie restriction is a dietary regimen capable of extending life span in a variety of multicellular organisms. A yeast model of calorie restriction has been developed in which limiting the concentration of glucose in the growth media of Saccharomyces cerevisiae leads to enhanced replicative and chronological longevity. Since S. cerevisiae are Crabtree-positive cells that present repression of aerobic catabolism when grown in high glucose concentrations, we investigated if this phenomenon participates in life span regulation in yeast. S. cerevisiae only exhibited an increase in chronological life span when incubated in limited concentrations of glucose. Limitation of galactose, raffinose or glycerol plus ethanol as substrates did not enhance life span. Furthermore, in Kluyveromyces lactis, a Crabtree-negative yeast, glucose limitation did not promote an enhancement of respiratory capacity nor a decrease in reactive oxygen species formation, as is characteristic of conditions of caloric restriction in S. cerevisiae. In addition, K. lactis did not present an increase in longevity when incubated in lower glucose concentrations. Altogether, our results indicate that release from repression of aerobic catabolism is essential for the beneficial effects of glucose limitation in the yeast calorie restriction model. Potential parallels between these changes in yeast and hormonal regulation of respiratory rates in animals are discussed.

Alkaline phosphatase as a reporter of sigma(S) levels and rpoS polymorphisms in different E-coli strains

sigma(S) is responsible for the transcriptional regulation of genes related to protection against stresses and bacterial survival and it accumulates in the cell under conditions of stress, such as nutrient limitation. An increase in the levels of sigma(S) causes a reduction in the expression of genes that are transcribed by RNA polymerase associated with the principal sigma factor, sigma(70). phoA, that encodes alkaline phosphatase (AP) is expressed under phosphate shortage conditions, and is also repressed by sigma(S). Here we show that in a Pi-limited chemostat, accumulation of rpoS mutations is proportional to the intrinsic level of sigma(S) in the cells. Acquisition of mutations in rpoS relieves repression of the PHO genes. We also devised a non-destructive method based on the rpoS effect on AP that differentiates between rpo(S+) and rpoS mutants, as well as between high and low-sigma(S) producers. Using this method, we provide evidence that sigma(S) contributes to the repression of AP under conditions of Pi excess and that AP variation among different strains is at least partly due to intrinsic variation in sigma(S) levels. Consequently, a simple and non-destructive AP assay can be employed to differentiate between strains expressing different levels of sigma(S) on agar plates.

Role of CcpA in Polyhydroxybutyrate Biosynthesis in a Newly Isolated Bacillus sp MA3.3

The ccpA gene was inactivated in the polyhydroxybutyrate (PHB)-producing strain Bacillus sp. MA3.3 in order to reduce glucose catabolite repression over pentoses and develop improved bacterial strains for the production of PHB from lignocellulosic hydrolysates. Mutant Bacillus sp. MSL7 Delta CcpA are unable to grow on glucose and ammonia as sole carbon and nitrogen sources, respectively. Supplementation of glutamate as the nitrogen source or the substitution of the carbon source by xylose allowed the mutant to partially recover its growth performance. RT-PCR showed that CcpA stimulates the expression of the operon (gltAB), responsible for ammonia assimilation via glutamate in Bacillus sp. MA3.3. Moreover, it was demonstrated that the supplementation of xylose or glutamate was capable of stimulating gltAB operon expression independently of CcpA. In PHB production experiments in mineral media, it has been observed that the glucose catabolite repression over the pentoses was partially released in MSL7. Although the carbohydrate consumption is faster in the ccpA mutant, the biomass and PHB biosynthesis are lower, even with supplementation of glutamate. This is attributed to an increase of acetyl-CoA flux towards the tricarboxylic acid cycle observed in the mutant. Copyright (C) 2011 S. Karger AG, Basel

Screening of bacteria to produce polyhydroxyalkanoates from xylose

Although xylose is a major constituent of lignocellulosic feedstock and the second most abundant sugar in nature, only 22% of 3,152 screened bacterial isolates showed significant growth in xylose in 24 h. Of those 684, only 24% accumulated polyhydroxyalkanoates after 72 h. A mangrove isolate, identified as Bacillus sp. MA3.3, yielded the best results in literature thus far for Gram-positive strains in experiments with glucose and xylose as the sole carbon source. When glucose or xylose were supplied, poly-3-hydroxybutyrate (PHB) contents of cell dry weight were, respectively, 62 and 64%, PHB yield 0.25 and 0.24 g g(-1) and PHB productivity (P(PHB)) 0.10 and 0.06 g l(-1) h(-1). This 40% P(PHB) difference may be related to the theoretical ATP production per 3-hydroxybutyrate (3HB) monomer calculated as 3 mol mol(-1) for xylose, less than half of the ATP/3HB produced from glucose (7 mol mol(-1)). In PHB production using sugar mixtures, all parameters were strongly reduced due to carbon catabolite repression. PHB production using Gram-positive strains is particularly interesting for medical applications because these bacteria do not produce lipopolysaccharide endotoxins which can induce immunogenic reactions. Moreover, the combination of inexpensive substrates and products of more value may lead to the economical sustainability of industrial PHB production.

Transcriptional Processing of the pst Operon of Escherichia coli

The pst operon of Escherichia coli is composed of five genes that encode a high-affinity phosphate transport system. pst belongs to the PHO regulon, which is a group of genes and operons that are induced in response to phosphate limitation. The pst operon also has a regulatory role in the repression of PHO genes` transcription under phosphate excess conditions. Transcription of pst is initiated at the promoter located upstream to the first gene, pstS. Immediately after its synthesis, the primary transcript of pst is cleaved into shorter mRNA molecules in a ribonuclease E-dependent manner. Other ribonucleases, such as RNase III and MazF, do not play a role in pst mRNA processing. RNase E is thus at least partially responsible for processing the pst primary transcript.

Ligand induced interaction of thyroid hormone receptor beta with its coregulators

Thyroid hormones exert most of their physiological effects through two thyroid hormone receptor (TR) subtypes, TR alpha and TR beta, which associate with many transcriptional coregulators to mediate activation or repression of target genes. The search for selective TR beta ligands has been stimulated by the finding that several pharmacological actions mediated by TR beta might be beneficial in medical conditions such as obesity, hypercholesterolemia and diabetes. Here, we present a new methodology which employs surface plasmon resonance to investigate the interactions between TR beta ligand binding domain (LBD) complexes and peptides derived from the nuclear receptor interaction motifs of two of its coregulators, SRC2 and DAX1. The effect of several TR beta ligands, including the TR beta selective agonist GC-I and the TR beta selective antagonist NH-3, were investigated. We also determined the kinetic rate constants for the interaction of TR beta-T3 with both coregulators, and accessed the thermodynamic parameters for the interaction with DAX1. Our findings Suggest that flexibility plays an important role in the interaction between the receptor and its coregulators. and point out important aspects of experimental design that should be addressed when using TR beta LBD and its agonists. Furthermore, the methodology described here may be useful for the identification of new TR beta ligands. (C) 2008 Elsevier Ltd. All rights reserved.

The Arabidopsis bZIP Gene AtbZIP63 Is a Sensitive Integrator of Transient Abscisic Acid and Glucose Signals

Glucose modulates plant metabolism, growth, and development. In Arabidopsis (Arabidopsis thaliana), Hexokinase1 (HXK1) is a glucose sensor that may trigger abscisic acid (ABA) synthesis and sensitivity to mediate glucose-induced inhibition of seedling development. Here, we show that the intensity of short-term responses to glucose can vary with ABA activity. We report that the transient (2 h/4 h) repression by 2% glucose of AtbZIP63, a gene encoding a basic-leucine zipper (bZIP) transcription factor partially involved in the Snf1-related kinase KIN10-induced responses to energy limitation, is independent of HXK1 and is not mediated by changes in ABA levels. However, high-concentration (6%) glucose-mediated repression appears to be modulated by ABA, since full repression of AtbZIP63 requires a functional ABA biosynthetic pathway. Furthermore, the combination of glucose and ABA was able to trigger a synergistic repression of AtbZIP63 and its homologue AtbZIP3, revealing a shared regulatory feature consisting of the modulation of glucose sensitivity by ABA. The synergistic regulation of AtbZIP63 was not reproduced by an AtbZIP63 promoter-5`-untranslated region:beta-glucuronidase fusion, thus suggesting possible posttranscriptional control. A transcriptional inhibition assay with cordycepin provided further evidence for the regulation of mRNA decay in response to glucose plus ABA. Overall, these results indicate that AtbZIP63 is an important node of the glucose-ABA interaction network. The mechanisms by which AtbZIP63 may participate in the fine-tuning of ABA-mediated abiotic stress responses according to sugar availability (i.e., energy status) are discussed.

The GATA-type transcriptional activator Gat1 regulates nitrogen uptake and metabolism in the human pathogen Cryptococcus neoformans

Nitrogen uptake and metabolism are essential to microbial growth. Gat1 belongs to a conserved family of zinc finger containing transcriptional regulators known as GATA-factors. These factors activate the transcription of Nitrogen Catabolite Repression (NCR) sensitive genes when preferred nitrogen sources are absent or limiting. Cryptococcus neoformans GAT1 is an ortholog to the Aspergillus nidulans AreA and Candida albicans GAD genes. In an attempt to define the function of this transcriptional regulator in C. neoformans, we generated null mutants (gat1 Delta) of this gene. The gat 1 mutant exhibited impaired growth on all amino acids tested as sole nitrogen sources, with the exception of arginine and proline. Furthermore, the gat1 mutant did not display resistance to rapamycin, an immunosuppressant drug that transiently mimics a low-quality nitrogen source. Gal is not required for C. neoformans survival during macrophage infection or for virulence in a mouse model of cryptococcosis. Microarray analysis allowed the identification of target genes that are regulated by Gat1 in the presence of proline, a poor and non-repressing nitrogen source. Genes involved in ergosterol biosynthesis, iron uptake, cell wall organization and capsule biosynthesis, in addition to NCR-sensitive genes, are Gat1-regulated in C. neoformans. (C) 2010 Elsevier Inc. All rights reserved.

Combates entre la memoria y la historia de Chile : Conflictos sobre el pasado reciente

The reestablishment of democracy in Chile has seen an intense debate about the events of the recent past, especially on the issue of human rights. From the very beginning, the Concertacion Government has been determined to discover the truth of the repression carried out by the national security forces with a series of commissions that have gathered the testimonies of victims and their relatives. These efforts have been resisted by conservative sectors linked to the dictatorship and the Armed Forces. There has been intense conflict in the media during the past 20 years about events that occurred during the rule of Salvador Allende and the Military Regime. In this regard, a great diversity of information has been produced which, together with the debate evoked, has enabled historians not only to rigorously and thoroughly reconstruct the operation of the state terror but also to explain how a significant sector of Chile’s civil society allowed that situation. This article presents, on one hand, different methodological tools in order to study the recent past and, on the other hand, the social discussion on how to do it.

Los asilados de las embajadas de Europa Occidental en Chile tras el golpe militar y sus consecuencias diplomáticas : El caso de Suecia

Given the expectations that western governments had in the reforms being implemented by the Unidad Popular, the September 1973 coup d’état in Chile caused a great international outcry. The violence and repression that accompanied the military coup produced a worldwide repudiation of the military junta. Sweden stood out most consistently among the countries that declared their opposition to Chile during the period of the dictatorship by continuously denouncing the violations to human rights. In this context, Sweden’s Ambassador to Chile, Harald Edelstam, played a very important role in saving the lives of hundreds of Chilean citizens and foreign nationals who sought refuge from the regime’s bloody repression. Making use of the privileges he enjoyed as a diplomat, Edelstam confronted the military authorities to release people wrongfully sentenced to the death penalty. As a result, he was declared persona non grata and forced to leave Chile. This was the beginning of a long and controversial diplomatic relationship that lasted until the return of democracy in Chile in 1990. This article is a first attempt at understanding how human rights violations affected bilateral relations between Chile and Sweden with respect to the diplomatic efforts of Ambassador Harald Edelstam.

A Psychoanalytical Reading of Emily Brontë's Wuthering Heights : An Analysis of the Defense Mechanisms of Some Characters

This essay presents a portrayal of Heathcliff, Catherine and Isabella from a psychoanalytical perspective with regard to four defense mechanisms; namely, repression, denial, sublimation and projection in order to see how these defense mechanisms have affected the characters’ decisions and behaviour, and led them to their destinations in life. It will include three major sections: repression in characters, denial in characters, and sublimation and projection in characters. These terms will be more clearly defined and explained in the subsequent sections.

Observatório de Inovação Social do Turismo: prevenção e enfrentamento da exploração sexual de crianças e adolescentes no turismo

The "Observatório de Inovação Social do Turismo" was created due to the need of studies and researches related to tourism and its social impacts. The Escola Brasileira de Administração Pública e de Empresas, of Fundação Getulio Vargas, in partnership with the Ministry of Tourism and the Childhood Brazil, started in 2007, studies about the sexual exploration of children and adolescents. The present work has the objective to analyze, based on the results of the "Observatório de Inovação Social do Turismo", its contributions to the prevention and repression of sexual exploration of children and adolescents associated to the tourism.

"A sala de detidos": atuação e ascenção da polícia política da capital federal do Brasil, 1920-1937

Exploramos neste trabalho o desenvolvimento e a ascensão da função de polícia política da capital federal do Brasil, aqui compreendida como um mecanismo de proteção e manutenção do poder do Estado. Tomando por base as décadas de 1920 e 1930, procuramos investigar duas hipóteses. Primeiro, que nessa primeira década, o exercício de polícia política, a cargo da 4ª. Delegacia Auxiliar, pode ser tomado como o início do aperfeiçoamento da função. Segundo que o decênio seguinte, ao ser marcado em seus primeiros anos por um processo de democratização das instituições brasileiras - que culmina com promulgação da Carta Constitucional de 1934, nos instiga à indagação sobre a necessidade do Estado contar oficialmente, a partir de 1933, com um órgão para o exercício exclusivo da função de polícia política. A análise contempla, a partir da legislação sobre reorganização dos serviços policiais e da repressão social e política, bem como acerca dos documentos produzidos pela polícia política, a relação entre o desenvolvimento desses órgãos e os eventos sociais e políticos no contexto nacional e internacional. Os pressupostos que nortearam o desenvolvimento dos órgãos de polícia política nas décadas enfocadas, sugerem que a relação de exclusividade entre os mesmos e os regimes autoritários deve ser problematizada, ainda que, ao longo da história, grande parte dos governos de matriz autoritária ou totalitária tenha desenvolvido órgãos de polícia política que exerceram um papel proeminente na estrutura de segurança do Estado. Longe, contudo, de esgotar a discussão, buscamos oferecer elementos adicionais para o entendimento sobre a necessidade de o Estado manter uma polícia voltada à produção de informações visando ao acompanhamento de certos eventos sociais e políticos.

Freud: da teoria psicanalítica à prática educacional

O emprego da teoria psicanalítica freudiana como fundamento para uma abordagem psicológica de problemas educacionais permite que se considere, simultaneamente, o desenvolvimento pleno do indivíduo e suas relações com a sociedade de que faz parte. Uma avaliação segura das possibilidades e limitações desse emprego exige um estudo sistemático da teoria, que inclui os instintos e suas vicissitudes - em especial repressão e sublimação -, o desenvolvimento da função sexual, o complexo de Édipo, a identificação, o superego e o narcisismo. Com base nesse estudo, pode-se afirmar que as idéias de Freud sobre educação concetram-se em dois processos que se prolongam por toda a vida do indivíduo: o desenvolvimento do ego e a integração à cultura. A aplicação do conhecimento psicanalítico à educação, de caráter profilático, apresenta grandes dificuldades. A proposta freudiana de uma educação para a realidade tem de ser posta em prática desde os primeiros anos da infância, o que exigiria urna problemática orientação a pais e professores. Entretanto, o emprego da psicanálise como disciplina científica ou procedimento de investigação para ampliar a compreensão do processo educacional poderá ter um caráter substancialmente renovador, desde que ela se integre à pesquisa interdisciplinar. Por essa via, sua contribuição será enriquecedora e ela própria se enriquecerá. De outro modo, revelar-se-á limitada para apreender o complexo conjunto das relações sociais.

O DOI-CODI carioca: memória e cotidiano no "Castelo do Terror"

O objetivo dessa dissertação é analisar a memória de seis ex-prisioneiros políticos do Destacamento de Operações de Informações-Centro de Operações de Defesa Interna do Rio de Janeiro (DOI-CODI/RJ), entrevistados recentemente, entre os anos de 2002 e 2004, sobre o cotidiano vivido nessa instituição em 1970. Naquele ano, dentro do Sistema de Segurança Interna (SISSEGIN), os DOI-CODI haviam sido criados e distribuídos por todas as Regiões Militares do país, tornando-se a principal instituição de repressão aos opositores políticos que optaram pela luta armada como forma de derrotar a ditadura militar brasileira. Assim, as narrativas desses seis ex-prisioneiros são, além de fontes essenciais, o principal objeto de estudo deste trabalho. Através delas, torna-se possível acessar aspectos cruciais para a caracterização do cotidiano vivido pelos presos em um desses órgãos, ― o DOI-CODI do Rio de Janeiro ―, uma vez que esse passado se liga ao presente por meio de suas memórias. Diante disso, a fim de melhor entender tais memórias, a formação e a atuação dos DOI-CODI também são aqui analisadas, colocando as narrativas dos ex-prisioneiros políticos entrevistados em diálogo com uma bibliografia especialmente selecionada, além de uma fonte a respeito do DOI feita por um de seus agentes quando este órgão ainda estava em atividade, em 1978. Para que a essas memórias seja aplicada uma crítica efetiva, necessária a todo trabalho histórico, o estudo se debruça ainda sobre as interferências que o presente exerce na construção que fazem com relação ao passado vivido no DOI-CODI/RJ, com o objetivo de esclarecer as bases sobre as quais são construídas cerca de trinta anos depois.