The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity

The putative eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein among archaea and eukaryotes that has recently been implicated in the elongation step of translation. eIF5A undergoes an essential and conserved posttranslational modification at a specific lysine to generate the residue hypusine. The enzymes deoxyhypusine synthase (Dys1) and deoxyhypusine hydroxylase (Lia1) catalyze this two-step modification process. Although several Saccharomyces cerevisiae eIF5A mutants have importantly contributed to the study of eIF5A function, no conditional mutant of Dys1 has been described so far. In this study, we generated and characterized the dys1-1 mutant, which showed a strong depletion of mutated Dys1 protein, resulting in more than 2-fold decrease in hypusine levels relative to the wild type. The dys1-1 mutant demonstrated a defect in total protein synthesis, a defect in polysome profile indicative of a translation elongation defect and a reduced association of eIF5A with polysomes. The growth phenotype of dys1-1 mutant is severe, growing only in the presence of 1 M sorbitol, an osmotic stabilizer. Although this phenotype is characteristic of Pkc1 cell wall integrity mutants, the sorbitol requirement from dys1-1 is not associated with cell lysis. We observed that the dys1-1 genetically interacts with the sole yeast protein kinase C (Pkc1) and Asc1, a component of the 40S ribosomal subunit. The dys1-1 mutant was synthetically lethal in combination with asc1Δ and overexpression of TIF51A (eIF5A) or DYS1 is toxic for an asc1Δ strain. Moreover, eIF5A is more associated with translating ribosomes in the absence of Asc1 in the cell. Finally, analysis of the sensitivity to cell wall-perturbing compounds revealed a more similar behavior of the dys1-1 and asc1Δ mutants in comparison with the pkc1Δ mutant. These data suggest a correlated role for eIF5A and Asc1 in coordinating the translational control of a subset of mRNAs associated with cell integrity. © 2013 Galvão et al.

Biodiversity Can Help Prevent Malaria Outbreaks in Tropical Forests

Background: Plasmodium vivax is a widely distributed, neglected parasite that can cause malaria and death in tropical areas. It is associated with an estimated 80-300 million cases of malaria worldwide. Brazilian tropical rain forests encompass host- and vector-rich communities, in which two hypothetical mechanisms could play a role in the dynamics of malaria transmission. The first mechanism is the dilution effect caused by presence of wild warm-blooded animals, which can act as dead-end hosts to Plasmodium parasites. The second is diffuse mosquito vector competition, in which vector and non-vector mosquito species compete for blood feeding upon a defensive host. Considering that the World Health Organization Malaria Eradication Research Agenda calls for novel strategies to eliminate malaria transmission locally, we used mathematical modeling to assess those two mechanisms in a pristine tropical rain forest, where the primary vector is present but malaria is absent. Methodology/Principal Findings: The Ross-Macdonald model and a biodiversity-oriented model were parameterized using newly collected data and data from the literature. The basic reproduction number (R0) estimated employing Ross-Macdonald model indicated that malaria cases occur in the study location. However, no malaria cases have been reported since 1980. In contrast, the biodiversity-oriented model corroborated the absence of malaria transmission. In addition, the diffuse competition mechanism was negatively correlated with the risk of malaria transmission, which suggests a protective effect provided by the forest ecosystem. There is a non-linear, unimodal correlation between the mechanism of dead-end transmission of parasites and the risk of malaria transmission, suggesting a protective effect only under certain circumstances (e.g., a high abundance of wild warm-blooded animals). Conclusions/Significance: To achieve biological conservation and to eliminate Plasmodium parasites in human populations, the World Health Organization Malaria Eradication Research Agenda should take biodiversity issues into consideration. © 2013 Laporta et al.

Validation of an alternative analytical method for the quantification of antioxidant activity in plant extracts

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development and validation of a D-loop mtDNA SNP assay for the screening of specimens in forensic casework

Mitochondrial DNA (mtDNA) analysis is usually a last resort in routine forensic DNA casework. However, it has become a powerful tool for the analysis of highly degraded samples or samples containing too little or no nuclear DNA, such as old bones and hair shafts. The gold standard methodology still constitutes the direct sequencing of polymerase chain reaction (PCR) products or cloned amplicons from the HVS-1 and HVS-2 (hypervariable segment) control region segments. Identifications using mtDNA are time consuming, expensive and can be very complex, depending on the amount and nature of the material being tested. The main goal of this work is to develop a less labour-intensive and less expensive screening method for mtDNA analysis, in order to aid in the exclusion of non-matching samples and as a presumptive test prior to final confirmatory DNA sequencing. We have selected 14 highly discriminatory single nucleotide polymorphisms (SNPs) based on simulations performed by Salas and Amigo (2010) [1] to be typed using SNaPShotTM (Applied Biosystems, Foster City, CA, USA). The assay was validated by typing more than 100 HVS-1/HVS-2 sequenced samples. No differences were observed between the SNP typing and DNA sequencing when results were compared, with the exception of allelic dropouts observed in a few haplotypes. Haplotype diversity simulations were performed using 172 mtDNA sequences representative of the Brazilian population and a score of 0.9794 was obtained when the 14 SNPs were used, showing that the theoretical prediction approach for the selection of highly discriminatory SNPs suggested by Salas and Amigo (2010) [1] was confirmed in the population studied. As the main goal of the work is to develop a screening assay to skip the sequencing of all samples in a particular case, a pair-wise comparison of the sequences was done using the selected SNPs. When both HVS-1/HVS-2 SNPs were used for simulations, at least two differences were observed in 93.2% of the comparisons performed. The assay was validated with casework samples. Results show that the method is straightforward and can be used for exclusionary purposes, saving time and laboratory resources. The assay confirms the theoretic prediction suggested by Salas and Amigo (2010) [1]. All forensic advantages, such as high sensitivity and power of discrimination, as also the disadvantages, such as the occurrence of allele dropouts, are discussed throughout the article. © 2013 Elsevier B.V.

Age-related differences in time-limit performance and force platform-based balance measures during one-leg stance

Poor posture control has been associated with an increased risk of falls and mobility disability among older adults. This study was conducted to assess the test-retest reliability and sensitivity to group differences regarding the time-limit (TLimit) of one-leg standing and selected balance parameters obtained with a force platform in older and young adults. A secondary purpose was to assess the relationship between TLimit and these balance parameters. Twenty-eight healthy older adults (age: 69±5years) and thirty young adults (age: 21±4years) participated in this study. Two one-leg stance tasks were performed: (1) three trials of 30s maximum and (2) one TLimit trial. The following balance parameters were computed: center of pressure area, RMS sway amplitude, and mean velocity and mean frequency in both the anterio-posterior and medio-lateral directions. All balance parameters obtained with the force platform as well as the TLimit variable were sensitive to differences in balance performance between older and young adults. The test-retest reliability of these measures was found to be acceptable (ICC: 0.40-0.85), with better ICC scores observed for mean velocity and mean frequency in the older group. Pearson correlations coefficients (r) between balance parameters and TLimit ranged from -0.16 to -0.54. These results add to the current literature that can be used in the development of measurement tools for evaluating balance in older and young adults. © 2013 Elsevier Ltd.

Search for methylation-sensitive amplification polymorphisms in mutant figs

Fig (Ficus carica) breeding programs that use conventional approaches to develop new cultivars are rare, owing to limited genetic variability and the difficulty in obtaining plants via gamete fusion. Cytosine methylation in plants leads to gene repression, thereby affecting transcription without changing the DNA sequence. Previous studies using random amplification of polymorphic DNA and amplified fragment length polymorphism markers revealed no polymorphisms among select fig mutants that originated from gamma-irradiated buds. Therefore, we conducted methylation-sensitive amplified polymorphism analysis to verify the existence of variability due to epigenetic DNA methylation among these mutant selections compared to the main cultivar 'Roxo-de-Valinhos'. Samples of genomic DNA were double-digested with either HpaII (methylation sensitive) or MspI (methylation insensitive) and with EcoRI. Fourteen primer combinations were tested, and on an average, non-methylated CCGG, symmetrically methylated CmCGG, and hemimethylated hmCCGG sites accounted for 87.9, 10.1, and 2.0%, respectively. MSAP analysis was effective in detecting differentially methylated sites in the genomic DNA of fig mutants, and methylation may be responsible for the phenotypic variation between treatments. Further analyses such as polymorphic DNA sequencing are necessary to validate these differences, standardize the regions of methylation, and analyze reads using bioinformatic tools. © FUNPEC-RP.

Selecting green suppliers based on GSCM practices: Using fuzzy TOPSIS applied to a Brazilian electronics company

Due to an increased awareness and significant environmental pressures from various stakeholders, companies have begun to realize the significance of incorporating green practices into their daily activities. This paper proposes a framework using Fuzzy TOPSIS to select green suppliers for a Brazilian electronics company; our framework is built on the criteria of green supply chain management (GSCM) practices. An empirical analysis is made, and the data are collected from a set of 12 available suppliers. We use a fuzzy TOPSIS approach to rank the suppliers, and the results of the proposed framework are compared with the ranks obtained by both the geometric mean and the graded mean methods of fuzzy TOPSIS methodology. Then a Spearman rank correlation coefficient is used to find the statistical difference between the ranks obtained by the three methods. Finally, a sensitivity analysis has been performed to examine the influence of the preferences given by the decision makers for the chosen GSCM practices on the selection of green suppliers. Results indicate that the four dominant criteria are Commitment of senior management to GSCM; Product designs that reduce, reuse, recycle, or reclaim materials, components, or energy; Compliance with legal environmental requirements and auditing programs; and Product designs that avoid or reduce toxic or hazardous material use. © 2013 Elsevier B.V. All rights reserved.

Planejamento da expansão de sistemas elétricos de distribuição considerando fatores de riscos em análise econômica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modernidade, medo e violência: reflexões teóricas e o caso de Marília/SP

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Planejamento econômico de gráficos de controle X para monitoramento de processos autocorrelacionados

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Monitoramento da solução do solo e do estado nutricional de laranjeiras Valência em cultivo fertirrigado

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Relação entre uniformidade da produtividade e indicadores de uniformidade da irrigação em sistema de irrigação por microaspersão em citros

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estimação de rigidezes de mancais de rotores por análise de sensibilidade

Pós-graduação em Engenharia Mecânica - FEIS

Estudo teórico-experimental em via de determinação de lei de atrito em escoamentos de fluidos hiperconcentrados

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

Análise da estabilidade estática de tensão de sistemas elétricos de potência usando uma rede neural baseada na teoria da ressonância adaptativa

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)