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.

Does bench model fidelity interfere in the acquisition of suture skills by novice medical students?

OBJETIVO: Embora vários modelos de bancada inanimados tenham sido descritos para o treinamento de habilidades de sutura, até o momento, não existe um método ideal para esse ensino e aprendizagem durante a formação médica. O objetivo foi avaliar se a fidelidade dos modelos de bancada interfere na aquisição de habilidades de sutura em estudantes de medicina iniciantes na prática cirúrgica. MÉTODOS: 36 estudantes de medicina sem exposição prévia a habilidades cirúrgicas foram randomizados em três grupos (n = 12): treinamento de suturas baseado em materiais didáticos (controle); treinamento de suturas em modelo de baixa-fidelidade (modelo de bancada de etileno vinil acetato); ou treinamento de suturas em modelo de alta-fidelidade (modelo de bancada de pele de pata de porco). Foram aplicados pré e pós-testes (realização de pontos simples e pontos subdérmicos invertidos em língua de boi). Três ferramentas (Global Rating Scale com avaliação cega, tamanho do efeito e autopercepção da confiança baseada em uma escala de Likert) foram utilizadas para mensurar todas as performances de sutura. RESULTADOS: A análise após o treinamento demonstrou que os estudantes que treinaram nos modelos tiveram um melhor (p < 0.0000) desempenho na avaliação pela Global Rating Scale, quando comparados com o controle, independente da fidelidade do modelo. A magnitude do efeito (treinamento) foi considerada grande (> 0.80) em todas as mensurações. Após o treinamento os alunos sentiram-se mais confiantes (p < 0.0000) para executarem os dois tipos de suturas. CONCLUSÃO: A aquisição de habilidades de suturas no modelo de baixa fidelidade foi semelhante à prática no modelo de alta fidelidade, sendo que a melhora no desempenho dos participantes que treinaram nesses dois modelos foi superior à aprendizagem baseada em materiais didáticos.

Conserved sequences in the β subunit of archaeal and eukaryal translation initiation factor 2 (elF2), absent from elF5, mediate interaction with elF2γ

The eukaryotic translation initiation factor 2 (eIF2) binds the methionyl-initiator tRNA in a GTP-dependent mode. This complex associates with the 40 S ribosomal particle, which then, with the aid of other factors, binds to the 5' end of the mRNA and migrates to the first AUG codon, where eIF5 promotes GTP hydrolysis, followed by the formation of the 80 S ribosome. Here we provide a comparative sequence analysis of the β subunit of eIF2 and its archaeal counterpart (aIF2β). aIF2β differs from eIF2β in not possessing an N-terminal extension implicated in binding RNA, eIF5 and eIF2B. The remaining sequences are highly conserved, and are shared with eIF5. Previously isolated mutations in the yeast eIF2β, which allow initiation of translation at UUG codons due to the uncovering of an intrinsic GTPase activity in eIF2, involve residues that are conserved in aIF2β, but not in eIF5. We show that the sequence of eIF2B homologous to aIF2β is sufficient for binding eIF2γ, the only subunit with which it interacts, and comprises, at the most, 78 residues, eIF5 does not interact with eIF2γ, despite its similarity with eIF2β, probably because of a gap in homology in this region. These observations have implications for the evolution of the mechanism of translation initiation.

Leucine-rich diet alters the eukaryotic translation initiation factors expression in skeletal muscle of tumour-bearing rats

Background: Cancer-cachexia induces a variety of metabolic disorders on protein turnorver, decreasing protein synthesis and increasing protein degradation. Controversly, insulin, other hormones, and branched-chain amino acids, especially leucine, stimulate protein synthesis and modulate the activity of translation initiation factors involved in protein synthesis. Since the tumour effects are more pronounced when associated with pregnancy, ehancing muscle-wasting proteolysis, in this study, the influence of a leucine-rich diet on the protein synthesis caused by cancer were investigated. Methods: Pregnant rats with or without Walker 256 tumour were distributed into six groups. During 20 days of experiment, three groups were fed with a control diet: C - pregnant control, W - tumour-bearing, and P - pair-fed, which received the same amount of food as ingested by the W group; three other groups of pregnant rats were fed a leucine-rich diet: L - pregnant leucine, WL - tumour-bearing, and PL - pair-fed, which received the same amount of food as ingested by the WL group. Results: The gastrocnemius muscle of WL rats showed increased incorporation of leucine in protein compared to W rats; the leucine-rich diet also prevented the decrease in plasma insulin normally seen in W. The expression of translation initiation factors increased when tumour-bearing rats fed leucine-rich diet, with increase of ∼35% for eIF2α and eIF5, ∼17% for eIF4E and 20% for eIF4G; the expression of protein kinase S6K1 and protein kinase C was also highly enhanced. Conclusion: The results suggest that a leucine-rich diet increased the protein synthesis in skeletal muscle in tumour-bearing rats possibly through the activation of eIF factors and/or the S6kinase pathway. © 2007 Ventrucci et al; licensee BioMed Central Ltd.

Translation, cross-cultural adaptation and validation of the Portuguese version of the DYMUS questionnaire for the assessment of dysphagia in multiple sclerosis

Translation, cross-cultural adaptation and validation of the DYMUS questionnaire for the assessment of dysphagia in multiple sclerosis. The original English version of the DYMUS was translated using the forward-backward technique, cross-culturally adaptated, pilot-tested in 40 patients, and then applied to 100 multiple sclerosis patients to assess the reliability and construct validity. Construct validity was assessed by Mann–Whitney test and Spearman’s correlation coefficient (rs). The internal consistency of the questionnaire was evaluated using Cronbach’s alpha coefficient and inter-item correlation. DYMUS-BR internal consistency was high (Cronbach’s alpha= 0.72); Cronbach’s alpha was 0.65 for the ‘dysphagia for solids’ subscale and 0.67 for the ‘dysphagia for liquids’ subscale and positive inter-item correlations was found between all items, except for weight loss question. Significant association (p<0.001) and correlation (rs = 0,357; p = 0.01) was found between DYMUS-BR and dysphagia self-assessment. The DYMUS-BR questionnaire maintained the characteristics of that originally described, demonstrating to be a reliable, valid, easy and consistent tool to be used by health professionals for preliminary selection of Brazilian MS patients who need more specific instrumental analyses of swallowing.

Health related quality of life measure in systemic pediatric rheumatic diseases and its translation to different languages: an international collaboration

Background: Rheumatic diseases in children are associated with significant morbidity and poor health-related quality of life (HRQOL). There is no health-related quality of life (HRQOL) scale available specifically for children with less common rheumatic diseases. These diseases share several features with systemic lupus erythematosus (SLE) such as their chronic episodic nature, multi-systemic involvement, and the need for immunosuppressive medications. HRQOL scale developed for pediatric SLE will likely be applicable to children with systemic inflammatory diseases.Findings: We adapted Simple Measure of Impact of Lupus Erythematosus in Youngsters (SMILEY (c)) to Simple Measure of Impact of Illness in Youngsters (SMILY (c)-Illness) and had it reviewed by pediatric rheumatologists for its appropriateness and cultural suitability. We tested SMILY (c)-Illness in patients with inflammatory rheumatic diseases and then translated it into 28 languages. Nineteen children (79% female, n= 15) and 17 parents participated. The mean age was 12 +/- 4 years, with median disease duration of 21 months (1-172 months). We translated SMILY (c)-Illness into the following 28 languages: Danish, Dutch, French (France), English (UK), German (Germany), German (Austria), German (Switzerland), Hebrew, Italian, Portuguese (Brazil), Slovene, Spanish (USA and Puerto Rico), Spanish (Spain), Spanish (Argentina), Spanish (Mexico), Spanish (Venezuela), Turkish, Afrikaans, Arabic (Saudi Arabia), Arabic (Egypt), Czech, Greek, Hindi, Hungarian, Japanese, Romanian, Serbian and Xhosa.Conclusion: SMILY (c)-Illness is a brief, easy to administer and score HRQOL scale for children with systemic rheumatic diseases. It is suitable for use across different age groups and literacy levels. SMILY (c)-Illness with its available translations may be used as useful adjuncts to clinical practice and research.

Una lapide in via Mazzini: os sofrimentos em Auschwitz e os desafios da tradução

Giorgio Bassani in his book Una lapide in via Mazzin seeks to show to the readers all the sufferings passed by Italians during the Second World War and emphasized those persists until today. Aiming to make an unprecedented translation of this tale, it is expected that the text provides us great experiences in the language and we could see the actual difficulties and differences with the Portuguese language, despite being considered these languages closed ones. The objective of this work is to produce a translation that allows us to transmit all anxieties and reflections suffered by the main character of the tale. After the war, the survivor of the concentration camp can go back to your city. He appears exactly at the moment is being placed a headstone on the synagogue wall in homage to Jews deported and killed in the camps. Given the gaunt figure of the man with his striped pajamas, the population is driven to rethink their own indifference and , in a way, its share of blame. Thus, the return should be celebrated is rejected and the figure of the survivor becomes a nuisance. The function of the headstone is actually and literally put a stone on the story. To achieve the proposed objective for this work, we analyzed all the chapters of the story Una lapide in via Mazzini and tried to find equivalents in Portuguese. However, we not ignored the traces of meaning that Giorgio Bassani invested in his work. Therefore, it was necessary to use both printed and virtual dictionaries. With the advancement of the translation work, we realized the difficulty of finding appropriate solutions to achieve the desired effect . The fidelity proposed by the author became increasingly difficult as our work progressed. In the course of all the discussions, it was concluded that , in fact , when it comes to translation, maintaining fidelity to the original text in a foreign language is a difficult and laborious process

Signs, translation, and life in the Bakhtin circle and in Welby's significs

“Significs” is provisionally defined by Welby (1911: vii) as the study of the nature of significance in all its forms and relationships, of its workings in all spheres of human life and knowledge. Considering “significs” as a movement highlighting significance, Welby explores the action of signs in life; and more than the Saussurean sign composed of signifier and signified, the sign as understood by Welby refers to meaning as generated through signs in motion. This notion of “significs” empowers the study of signs when it considers the sign not in terms of the Saussurean structural representation of the union of the concept and acoustic image, but as (responsive and responsible) sign action in the world, in life. This also means to take into account the “extra-linguistic referent” (translinguistic and transdiscursive character of significs), history (space-time), subjectivity, the architecture of values connected to language, their communicative function. We believe that a dialogue can be established between Welby’s vision of significs and the notion of ideological sign proposed by Vološinov in Marxism and the Philosophy of Language, expanding the notions of “meaning” and “sense.”

Geographical variation in mutualistic networks: similarity, turnover and partner fidelity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)