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.

Cruzain inhibition by hydroxymethylnitrofurazone and nitrofurazone: investigation of a new target in Trypanosoma cruzi

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

Avaliação da atividade fotoprotetora de Achillea millefolium L. (Asteraceae)

As radiações ultravioleta (divididas em ultravioleta A, B e C), por possuírem diferentes energias e comprimentos de onda, apresentam diferentes atividades sobre os organismos, sendo que algumas destas atividades são benéficas e outras, não. A fim de evitar os malefícios causados por estas radiações, existem produtos denominados fotoprotetores. Uma vez que os flavonóides são considerados um fator importante de proteção contra a radiação ultravioleta, alguns estudos concentram-se na avaliação da atividade fotoprotetora que algumas plantas, compostas por flavonóides, possam apresentar. Sendo assim, este trabalho teve como objetivo avaliar a atividade fotoprotetora de extratos das flores e folhas de Achillea millefolium L. através de testes in vitro por espectrofotometria no ultravioleta. Embora a literatura descreva a presença de flavonóides nas flores e folhas de A. millefolium, não foi encontrada atividade fotoprotetora nos extratos testados.

Avaliação de métodos de análises para determinação da contagem de células somáticas no leite cru, mantido em tanque de resfriamento

We analyzed the quality of raw milk from eight dairy farms in Rio Grande do Norte stored in a cooling tank , in order to evaluate methods for determining somatic cell counts (SCC). The Somaticell® kit and a portable Direct Cell Counter (DCC) were compared with each other and with the MilkoScanTM FT+ (FOSS Denmark), which uses Fourier Transform Infrared (FTIR) spectroscopy). Direct cell counter data were processed for somatic cell scores (log-transformed somatic cell count) and analyzed with the SAS®, statistical package , Statistical Analysis System, (SAS, INSTITUTE, 1998). Comparison of means and correlation of somatic cell scores were conducted using Pearson s correlation coefficient and the Tukey Test at 1 %. No significant difference was observed for comparison of means. The correlation between somatic cell scores was significant, that is, 0.907 and 0.876 between the MilkoScanTM FT+ and the Somaticell® kit and Direct Cell Count (DCC) respectively, and 0.943 between the Somaticell® kit and Direct Cell Count (DCC). The methods can be recommended for monitoring the quality of raw milk kept in a cooling tank in the production unit

Fatores de influência na produção de leite bovino sobre os níveis da contagem bacteriana total

The objective of this study was to evaluate the influence of milking procedures on the levels of total bacterial count (TBC) in bovine milk. In the first study the influences of procedures for hygienic milking, cleaning of milking equipment and milk cooling tanks on the TBC levels were evaluated. Four bulk samples of milk were collected from each tank in eight properties for TBC analysis, employing the flow cytometry method. A questionnaire was applied in each property to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology for analysis of longitudinal data was considered, focusing on random effects models. The results showed that the handling procedures for milking and the cleanliness of the cooling tank contributed to a further reduction in the levels of TBC raw milk cooling tanks. The second study aimed to describe the percentage of the properties that comply with the Normative Instruction Nº 51 (Brazil s IN 51) with regard to total bacterial count (TBC) in bovine milk. The study was conducted from January 2010 to July 2011. Milk samples were collected from the eight properties selected for TBC analysis by the flow cytometry method. Again, on each property a questionnaire was applied to assess the current situation of milking procedures on each production system that took part on this research, followed by training of employees in good agricultural practices in the production of milk and monitoring of the TBC measurements. The methodology of marginal models based on Generalized Estimate Equations (GEEs) was followed in the statistical analysis. The results showed that the handling procedures of the milking and the cleanliness of the cooling tanks contributed to a considerable percentage of the properties that reached the limits of TBC established by IN 51

Avaliação dos impactos ambientais e sociais gerados pela utilização do kit Embrapa de ordenha manual na produção de leite de cabra

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Caracterização dos sistemas de produção de leite bovino na microrregião Seridó do estado do Rio Grande do Norte

The objective was to contribute to a reading of the characteristics and diversity of production systems from bovine milk in the microregion Seridó of Rio Grande do Norte, discuss the social, economic, environmental and husbandry-related primary activity of milk production. We randomly selected 28 agricultural establishments that performed the activity of dairy bovine culture with subsequent applicat ion of a structured questionnaire during September and October 2011. Data were analyzed with application of measures of descriptive analysis and determination of the index rural development (IRD). The results showed that 53.57% of the interviewees were owners of the land, the median area of the properties amounted to 135 hectares, the median number of animals in the herd was 51 head, with minimum 11 and maximum of 350 heads establishments in the sample, 85.72% of establishments had maximum 23 cows in lactation, 100% performed manual milking with suckling calves, average productivity of 3.91 liters / cow / day, 92.86% of the interviewees did not produce silage and / or hay, 64.29% had no access to technical assistance, the average age of interviewees was 51 ± 10.85 years, 78.57% had only elementary education. The average of the IRD computed in the test sample amounted to 0.43 ± 0.11 on a scale 0-1. The sites were grouped into 04 classes, based on your IRD. The average IRD in each dimension was 0.43, 0.55 and 0.34, respectively for economic / husbandry, social and environmental. The determination of the rate of rural development - IDR found for the sample could add establishments hierarchically. It is necessary that the inclusion of best practices in herd management, bookkeeping zootechnical, technical assistance and reorganization practices in land and environmental preservation

Caracterização genética de vacas leiteiras por meio de marcadores moleculares e suas implicações na composição e qualidade do leite

The objective of this study was to identify DNA polymorphisms at the genes leptin, β-lactoglobulin and pituitary-specific transcription factor in three genetic groups of Holstein x Guzerat dairy cows and investigate the relationship between their genotypes and the composition and quality of milk of dairy cows. Samples were collected in August 2009, being 113 blood samples from lactating crossbred cows and 58 milk samples. For analysis of DNA polymorphisms blood samples were collected, analyzed later in the Genetic Laboratory affiliated to the Zootechny Institute of São Paulo and individual milk samples were collected according to standards established by the laboratory of Management Program of Northeast Dairy Herds (PROGEN), at Federal Rural University of Pernambuco (UFRPE) for analysis of milk composition and quality. The characterization of genotypes was performed by PCR-RFLP, for which were designed specific primers for each studied gene and restriction enzymes Kpn2I, HaeIII and HinfI that cut the DNA of the following genes: leptin, β-lactoglobulin and a PIT, respectively. The leptin estimate genotypic frequence were CC 0.112, TT 0.225 and CT 0.661, for β-lactoglobulin were AA 0.136, AB 0.323 and BB 0.539, and for PIT were ++ 0.655, -- 0.311 and +- 0.032. The results show that the population is in Hardy-Weinberg disequilibrium for leptin, β-lactoglobulin and a PIT due to excess of heterozygotes in the population, however, as these genes are associated with the milk production it is considered that the animals have genetic potential for milk production in the Brazilian semi-arid conditions. Through the characterization of the studied herd there were not found implications of the polymorphism of leptin, β-lactoglobulin and PIT in the composition and quality of milk from cows in the different genetic groups 1/2, 3/4 and 7/8 Holstein x Guzerat. Key words: β-lactoglobulin, crossbred cows, leptin, PCR-RFLP, PIT1, semi-arid.

Características da carcaça e qualidade da carne de ovinos terminados em pastagens cultivadas

The objective of this study was to evaluate the quality of housing and the physical and chemical characteristics of meat from sheep raised on pasture Brachiaria brizantha and Panicum maximum. The experiment was conducted in the physical area of the Study Group on Forage (GEFOR), located in the Academic Unit Specialized in Agricultural Sciences - Federal University of Rio Grande do Norte - UFRN in Macaíba, RN, Brazil. We used 32 lambs SPRD, obtained from herds in the state, with liveweight (LW) of 24.5 kg were assigned randomly to four treatments consisting of tropical grasses, two cultivars of Brachiaria brizantha, Marandu and Piatã, and two of Panicum maximum, Aruana and Massai. The experimental area was 2.88 ha, divided into 4 paddocks of 0.72 ha, where each picket consisted of a farm and was divided into six plots of 0.12 ha, where the animals remained under rotational grazing. The period of adaptation to the pickets was seven days. At the beginning of the experiment the animals were weighed, identified with plastic earrings and necklaces colored according to the treatment, and treated against. The lambs were loose in the paddock at 8 am and collected at 16 hours, which returned to collective pens. During the time of grazing animals had free access to mineral supplement with monensin Ovinofós ® and water. Before entering the paddocks of pasture were sampled to characterize the chemical composition. Every seven days occurred at weighing, with fasting, to monitor the weight development. Cultivars Marandu, Aruana, Piatã and Massai were grazed for 133, 129, 143 and 142 days, respectively, until the lambs reach slaughter weight. Arriving at 32 kg lambs were evaluated subjectively for body condition score by, passed through fasting period, diet and water for 16 hours were slaughtered. Measurements were made in the inner and outer casings in addition to subjective evaluations regarding muscling, finish and quantity of pelvic-renal fat, then each was divided longitudinally into two half-carcases and cuts were made in the commercial left half, and after heavy calculated their income. Between the 12th and 13th thoracic vertebrae, was performed a cut to expose the cross section of the Longissimus dorsi, which was drawn on the rib eye area (REA) in transparent film. Fat thickness and extent of AOL GR were determined using a caliper. A tissue composition was determined by dissection of the legs. Analyzes were performed physical (color, cooking loss and shear force) and chemical composition of meat (moisture, ash, protein and lipids) in Longissimus dorsi muscle. Grazing tropical grass Brachiaria brizantha cvs. Marandu and Piatã and Panicum maximum cvs. Aruana and Massai can be used for lambs SRPD in the rainy season, because not alter the physico-chemical and chemical composition of meat

Sistema de produção misto de caprino na região central do estado do Rio Grande do Norte

This study aimed to evaluate the performance and profitability of Canindé goats kept under semi-intensive management in mix farming system. 84 animals were used, being 81 does e three bucks.. The means of fertility rate at parturition, prolificacy and percentages of pregnant, kid survival and reproductive rate were compared by Tukey test at significance level of 5.0%. The kid weights from birth to weaning were submitted to variance analysis considering sex and birth type and the means were compared by Tukey test at 5.0% of significance. The percentages of fertility rate by ultrassographic were 91.36 and parturition 74.84. The prolificacy was 1.53. Independently of sex the mortality rate of kids from birth to weaning was 12.3. The average weights of offspring were 2.11 and 1.92 kg and 6.74 and 6.84 kg for male and female offspring at birth and weaning, in this order. The average weights of kids, meaning PN, P28, P56, P63, in the first season of kidding were 2.11 ± 0.46 kg, 3.63 ± 1.04 kg, 5.96 ± 158 kg and 6.74 ± 1.79 kg. There was significant differences (P<0,05) only to birth weight of kids sons of buck 135. However, the fertility at parturition for first and second breeding seasons (BS) were 87.65 and 62.96, respectively (P<0.05). The prolificacy was 1.58 and 1.47 to first and second BS in this order. The percentages of offspring survival from birth to weaning, to first and second BS were 92.86 and 80.00, respectively. From the first BS the reproduction rate was 1.28 while in the second was 0.74. After weaning the duration of lactation was 104 days to first breeding parturition and 38 days in the second one. The total milk production was 853.50 kg. The kids weight gain was influenced by season of year, wet and dry and buck (P<0.05). Were concluded that the seasons of year when breeding season and parturition occur strongly affected the productive performance and profitability of production system

Levantamento exploratório da composição e frações proteicas do leite bovino em propriedades leiteras

The direct relationship with the processing, industrial yield and price of milk, the importance of the centesimal composition of milk and their protein fractions has increased for both the dairy industry and for the producers, so it is important to identify the possible causes on variation of these components. The aim of this study was to evaluate the effect of the seasons, order of calving and lactation phase on milk composition and protein fractions in dairy farms in the Agreste and Leste meso-regions, of Rio Grande do Norte State (RN). Milk samples were taken in seven milk farms directly from the bulk tanks except for one farm where samples were taken from higher milking cows producers and heifers. The experiment was done in seven properties with milk samples taken directly from the bulk tanks and in one property were collected from higher production cows and heifers. Composition analyses were carried out for both experiments and sensorial trial was performed for the second experiment only. The data results were submitted to analysis of variance (ANOVA) and Tukey test performed for 5% significance level. In the first experiment was observed response of the seasons, dry and rainy season; while in the second, the effect of the order of calving and lactation phase had influence on milk composition and protein fractions. There was significant response (p < 0.05) for season effects but due to farms. The rainy season had higher averages in the protein, lactose, total solids and nonfat dry extract. Multiparous cows had higher CCS, NUL and PCAS milk contents; while, first calving heifers showed higher average for the sensory attribute, color of milk. The final third of lactation was responsible for the elevation of the levels of almost all milk components and their protein fractions

Valor nutritivo de fenos de moringa (moringa oleifera lam) com diferentes idades de corte

This study was conducted to evaluate the consumption and digestibility of dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE), total carbohydrates (TC), non-fiber carbohydrates (NFC) and neutral detergent fiber (NDF) in sheep fed hay moringa (Moringa oleifera Lam) obtained with four cutting ages (28, 35, 42 and 49 days). We used 20 females Morada Nova breed, with 20kg of live weight, distributed in a completely randomized design and maintained in metabolism cages. There was a negative linear effect of age of cutting on DM intake, with an estimated maximum consumption of 0.67 kg / day for the hay produced at 28 days of cutting. It was also observed linear behavior, with an estimated maximum consumption 172g/dia, 0.36 kg / day; 18g/dia, at 35, 42 and 49 days old, for CP, OM and EE, respectively. For NDF, quadratic effect was found with advancing maturity of the plant. The apparent digestibility of DM, CP, OM and TC linearly decreased with advancing age of cutting hay Moringa oleifera. It was concluded that the hay Moringa oleifera showed better nutritional value after 28 days of cutting

Caracterização e modelagem dos sistemas de produção de caprinos leiteiros

The objectives of this research were characterizing the dairy goat production systems and model it using linear program. On the first step of this research, the model was developed using data from farms that was affiliated in the ACCOMIG/Caprileite, used a similar dairy goat production systems and have a partnership program with Universidade Federal de Minas Gerais . The data of research were from a structured questionnaire applied with farmers and monitoring of production systems during a guided visit on their farms. The results permitted identify that all farms were classified as a small and have a intensive production system. The average herd size had 63.75 dairy goats on lactation; it permits a production of 153, 38 kg of goat milk per day. It was observed that existing more than one channel of commercialization for the goat milk and their derivative products. The data obtained, on the first step of this research, was used to develop a linear program model. It was evaluated in two goat production systems, called P1 and P2. The results showed that the P1 system, with an annual birth and lactation during approximately 300 days was the best alternative for business. These results were compared with a mixed (beef and dairy) goat system in the semiarid region, which indicated merged with both systems. Therefore, to achieve profits and sustainability of the system, in all simulations it was necessary a minimum limit of funding of U.S. $ 10,000.00; this value permit earning of U.S. $ 792.00 per month and pay the investment within 5 years

Nanoindentation mechanical properties characterization of glassy polymeric carbon treated with ion beam

Phenolic resins when heat treated in inert atmosphere up to 1000 degreesC become glassy polymeric carbon (GPC), a chemically inert and biocompatible material useful for medical applications, such as in the manufacture of heart valves and prosthetic devices. In earlier work we have shown that ion bombardment can modify the surface of GPC, increasing its roughness. The enhanced roughness, which depends on the species, energy and fluence of the ion beam, can improve the biocompatibility of GPC prosthetic artifacts. In this work, ion bombardment was used to make a layer of implanted ions under the surface to avoid the propagation of microcracks in regions where cardiac valves should have pins for fixation of the leaflets. GPC samples prepared at 700 and 1500 degreesC were bombarded with ions of silicon. carbon, oxygen and gold at energies of 5, 6, 8 and 10 MeV, respectively, and fluences between 1.0 x 10(13) and 1.0 x 10(16) ions/cm(2). Nanoindentation hardness characterization was used to compare bombarded with non-bombarded samples prepared at temperatures up to 2500 degreesC. The results with samples not bombarded showed that the hardness of GPC increases strongly with the heat treatment temperature. Comparison with ion bombarded samples shows that the hardness changes according to the ion used, the energy and fluence. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Amorphous carbon nitrogenated films prepared by plasma immersion ion implantation and deposition

In this work, an investigation was conducted on amorphous hydrogenated-nitrogenated carbon films prepared by plasma immersion ion implantation and deposition. Glow discharge was excited by radiofrequency power (13.56 MHz, 40 W) whereas the substrate-holder was biased with 25 kV negative pulses. The films were deposited from benzene, nitrogen and argon mixtures. The proportion of nitrogen in the chamber feed (R-N) was varied against that of argon, while keeping the total pressure constant (1.3 Pa). From infrared reflectance-absorbance spectroscopy it was observed that the molecular structure of the benzene is not preserved in the film. Nitrogen was incorporated from the plasma while oxygen arose as a contaminant. X-ray photoelectron spectroscopy revealed that N/C and O/C atomic ratios change slightly with R-N. Water wettability decreased as the proportion of N in the gas phase increased while surface toughness underwent just small changes. Nanoindentation measurements showed that film deposition by means of ion bombardment was beneficial to the mechanical properties of the film-substrate interface. The intensity of the modifications correlates well with the degree of ion bombardment. (c) 2006 Elsevier B.V. All rights reserved.