Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis

Eukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal alpha-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins - eIF5A(K56A) and eIF5A(Q22H,L93F)- and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression.

Thioredoxin-1 promotes survival in cells exposed to S-nitrosoglutathione: Correlation with reduction of intracellular levels of nitrosothiols and up-regulation of the ERK1/2 MAP Kinases

Accumulating evidence indicates that post-translational protein modifications by nitric oxide and its derived species are critical effectors of redox signaling in cells. These protein modifications are most likely controlled by intracellular reductants. Among them, the importance of the 12 kDa dithiol protein thioredoxin-1 (TRX-1) has been increasingly recognized. However, the effects of TRX-1 in cells exposed to exogenous nitrosothiols remain little understood. We investigated the levels of intracellular nitrosothiols and survival signaling in HeLa cells over-expressing TRX-1 and exposed to S-nitrosoglutahione (GSNO). A role for TRX-1 expression on GSNO catabolism and cell viability was demonstrated by the concentration-dependent effects of GSNO on decreasing TRX-1 expression, activation of capase-3, and increasing cell death. The over-expressaion of TRX-1 in HeLa cells partially attenuated caspase-3 activation and enhanced cell viability upon GSNO treatment. This was correlated with reduction of intracellular levels of nitrosothiols and increasing levels of nitrite and nitrotyrosine. The involvement of ERK, p38 and JNK pathways were investigated in parental cells treated with GSNO. Activation of ERK1/2 MAP kinases was shown to be critical for survival signaling. lit cells over-expressing TRX-1, basal phosphorylation levels of ERK1/2 MAP kinases were higher and further increased after GSNO treatment. These results indicate that the enhanced cell viability promoted by TRX-1 correlates with its capacity to regulate the levels of intracellular nitiosothiols and to up-regulate the survival signaling pathway mediated by the ERK1/2 MAP kinases.

Co-stressors chilling and high light increase photooxidative stress in diuron-treated red alga Kappaphycus alvarezii but with lower involvement of H(2)O(2)

Diuron is one of the most commonly found N-phenylurea herbicides in marine/estuarine waters that promotes toxic effects by inhibiting photosynthesis and affecting the production of reactive oxygen species (ROS) in autotrophs. Since photo- and thermoacclimation are also ROS-mediated processes, this work evaluates a hypothetical additive effect of high light (HL) and chilling (12 degrees C) on 50 nM diuron toxicity to the highly-photosynthetically active apices of the red alga Kappaphycus alvarezii. Additive inhibition of photosynthesis was mainly evidenced by significant decreases of quantum yield of photosystem II and electron transfer rates upon co-stressors exposure to diuron-treated algae. Under extreme 12 degrees C/HL/diuron conditions, unexpected lower correlations between H(2)O(2) concentrations in seawater and radical-sensitive protein thiols were concomitantly measured with the highest indexes of photoinhibition (parameter beta). Altogether, these data support the hypothesis that co-stressors chilling/HL additively inhibit photosynthesis in diuron-exposed K. alvarezii but with less involvement of H(2)O(2) in injury effects than with only chilling or HL. (C) 2010 Elsevier Inc. All rights reserved.

Proteomic analysis of dorsolateral prefrontal cortex indicates the involvement of cytoskeleton, oligodendrocyte, energy metabolism and new potential markers in schizophrenia

Schizophrenia is likely to be a consequence of serial alterations in a number of genes that, together with environmental factors, will lead to the establishment of the illness. The dorsolateral prefrontal cortex (Brodmann`s Area 46) is implicated in schizophrenia and executes high functions such as working memory, differentiation of conflicting thoughts, determination of right and wrong concepts, correct social behavior and personality expression. We performed a comparative proteome analysis using two-dimensional gel electrophoresis of pools from 9 schizophrenia and 7 healthy control patients` dorsolateral prefrontal cortex aiming to identify, by mass spectrometry, alterations in protein expression that could be related to the disease. In schizophrenia-derived samples, our analysis revealed 10 downregulated and 14 upregulated proteins. These included alterations previously implicated in schizophrenia, such as oligodendrocyte-related proteins (myelin basic protein and transferrin), as well as malate dehydrogenase, aconitase, ATP synthase subunits and cytoskeleton-related proteins. Also, six new putative disease markers were identified, including energy metabolism, cytoskeleton and cell signaling proteins. Our data not only reinforces the involvement of proteins previously implicated in schizophrenia, but also suggests new markers, providing further information to foster the comprehension of this important disease. (C) 2008 Elsevier Ltd. All rights reserved.

Children's Vocabulary Development : The role of parental input, vocabulary composition and early communicative skills

The aim of this thesis is to examine the early vocabulary development of a sample of Swedish children in relation to parental input and early communicative skills. Three studies are situated in an overall description of early language development in children. The data analyzed in the thesis was collected within a larger project at Stockholm University (SPRINT- “Effects of enhanced parental input on young children’s vocabulary development and subsequent literacy development” [VR 2008-5094]). Data analysis was based on parental report via SECDI, the Swedish version of the MacArthur-Bates Communicative Development Inventories, and audio recordings. One study examined parental verbal interaction characteristics in three groups of children with varying vocabulary size at 18 months. The stability of vocabulary development at 18 and 24 months was investigated in a larger study, with focus on children’s vocabulary composition and grammatical abilities. The third study examined interrelations among early gestures, receptive and productive vocabulary, and grammar measured with M3L, i.e. three longest utterances, from 12 to 30 months. Overall results of the thesis highlight the importance of early language development. Variability in different characteristics in parental input is associated with variability in child vocabulary size. Children with large early vocabularies exhibit the most stability in vocabulary composition and the earliest grammatical development. Children’s vocabulary composition may reflect individual stylistic variation. Use of early gestures is associated differentially with receptive and productive vocabulary. Results of the thesis have implications for parents, child- and healthcare personnel, as well as researchers and educational practitioners. The results underscore the importance of high quality in adult-child interaction, with rich input fine-tuned to children’s developmental levels and age, together with high awareness of early language development.

Involvement of free radicals in peroxidatic reactions catalyzed by chloroperoxidase

The mechanism of chloroperoxidase (CPO)-catalyzed peroxidatic reactions of several substituted hydroquinones was studied at various hydrogen peroxide concentrations. The pathway was studied using cytochrome c as the radical trapping agent. As the hydroquinones became more hindered there was a difference in the amount of radicals trapped. For hydroquinone, 59.3% radical pathway, and methylhydroquinone, 81.4% radical, the difference in radicals trapped is due to a difference in pathway. For 2,3-dimethylhydroquinone (75.4%), trimethylhydroquinone (44.5%), and t-butylhydroquinone (0%) other non-peroxidatic reactions are noticed. Thus, for the more substituted hydroquinones the difference in radicals trapped can not be assigned to a difference in radical pathway. Also, there were problems drawing conclusions for this system due to the catalytic reaction of hydrogen peroxide. The radical trapping ability of 2,4,6-trimethylphenol was investigated for various other substrates. TMP reacted with the radicals generated in the enzymatic reactions of phenol, resorcinol, and m-methoxyphenol. Thus, this TMP system offers further potential as another radical trapping agent for use in these studies.

Avaliação da aplicabilidade da escala New Involvement Profile para mensuração do envolvimento do consumidor na cidade de Porto Alegre

O estudo do envolvimento do consumidor representa um dos tópicos mais importantes de pesquisa para aqueles que visam entender a forma como os seres humanos realizam suas decisões de compra. O envolvimento determina a quantidade de esforços empregados no processo de tomada de decisão, e apresenta importantes implicações tanto para o âmbito acadêmico quanto para o meio gerencial. No que diz respeito ao ambiente brasileiro, no entanto, foram poucos os trabalhos que se dedicaram exclusivamente a esse tema, não havendo, até o início desta pesquisa, nenhuma escala para mensuração de envolvimento validada para este cenário. O objetivo deste estudo é a avaliação da aplicabilidade da escala de Jain & Srinivasan (1990) na cidade de Porto Alegre. Tal escala, denominada New Involvement Profile (NIP), pode ser utilizada para mensuração do envolvimento de consumidores com diferentes categorias de produtos. Para esta pesquisa, mensurou-se o envolvimento com relação aos produtos cerveja, chocolate, corte de cabelo, CD, aparelho de televisão, detergente, shampoo e pilha. A escala NIP foi reaplicada a uma amostra de 420 (quatrocentos e vinte) respondentes em diversos bairros desta cidade. A qualidade e a robustez da escala de Jain & Srinivasan (1990) foram comprovadas através de testes de confiabilidade e verificações da validade de conteúdo e de constructo, englobando validade convergente e discriminante de cada dimensão da escala. Tal avaliação foi complementada a partir da relação existente entre os níveis de envolvimento encontrados e os pressupostos teóricos acerca desse conceito – tipos e determinates, bem como da comparação entre esses níveis e aqueles obtidos pelos mesmos produtos em estudos semelhantes.

Estilo parental percebido e adaptação psicológica de adolescentes adotados

Nas últimas décadas, diversos autores têm indicado que a condição de ser filho adotivo implica maior risco de desadaptação psicológica. Frente a isto, esta pesquisa investigou as relações existentes entre auto-estima, depressão, estilo parental percebido e adoção. A amostra foi composta por 524 adolescentes entre 14 e 15 anos de idade (68 adotados e 456 criados pelas famílias biológicas). Os instrumentos utilizados foram um questionário demográfico, as Escalas de Responsividade e Exigência Parental, o CDI e a Escala de Auto-Estima de Rosenberg. Análises de Regressão apontaram que as variáveis que apresentaram maior efeito sobre os índices de saúde emocional foram a responsividade parental, o sexo e o tipo de filiação. Os achados indicaram que pais adotivos são significativamente mais indulgentes do que pais biológicos. Em comparação, pais biológicos foram descritos por seus filhos como mais negligentes. Os resultados demonstraram ainda que a adoção isoladamente não resulta em maior depressão entre os jovens, mas a interação da afiliação com diversos outros fatores determina diferenças nestes escores. Os achados corroboraram o efeito transcultural dos estilos parentais sobre a adaptação psicológica e confirmaram a hipótese de que as estratégias de socialização parental moderam o desenvolvimento dos adolescentes adotados.

The stakeholders‟ involvement in the process of building and maintaining a destination brand

A destination is a place that attracts visitors for a temporary stay to participate in tourism related activities or non- activities. Globalization, the increased number of travelers and the increased buying power have increased the competition between the destinations and the destinations have become more substitutable. It has been agreed that destinations can be branded as well as products and to be competitive it is getting common to brand destinations. Destination Marketing Organizations (DMOs) are responsible for the marketing of an identifiable destination. The purpose of this study is to present an exploratory study of how a destination marketing organization creates and builds a strong destination brand and how the stakeholders have been involved in the process. The study is done with a qualitative case study approach. The case study was chosen as the research method to make a detailed and intensive analysis of the research objective, in this case the destination brand of Brazil and its stakeholders.

Cyclicality of SME Lending and Government Involvement in Banks

Recent regulatory efforts aim at lowering the cyclicality of bank lending because of its potential detrimental effects on financial stability and the real economy. We investigate the cyclicality of SME lending by local banks with vs. without a public mandate, controlling for location, size, loan maturity, funding structure, liquidity, profitability, and credit demand-side factors. The public mandate is set by local governments and stipulates a deviation from strict profit maximization and a sustainable provision of financial services to local customers. We find that banks with a public mandate are 25 percent less cyclical than other local banks. The result is credit supply-side driven and especially strong for savings banks with high liquidity and stable deposit funding. Our findings have implications for the banking structure, financial stability and the finance-growth nexus in a local context.

Cyclicality of SME lending and government involvement in banks

Recent regulatory efforts aim at lowering the cyclicality of bank lending because of its potential detrimental effects on financial stability and the real economy. We investigate the cyclicality of SME lending by local banks with vs. without a public mandate, controlling for location, size, loan maturity, funding structure, liquidity, profitability, and credit demand-side factors. The public mandate is set by local governments and stipulates a deviation from strict profit maximization and a sustainable provision of financial services to local customers. We find that banks with a public mandate are 25 percent less cyclical than other local banks. The result is credit supply-side driven and especially strong for savings banks with high liquidity and stable deposit funding. Our findings have implications for the banking structure, financial stability and the finance-growth nexus in a local context.