Eletroforese capilar acoplada à espectrometria de massas (CE-MS): vinte anos de desenvolvimento

CE-MS has been increasingly used for analysis of a vast array of compounds. This article reviews the different electrophoretic modes, interfaces and mass analyzers that are commonly used in the CE-MS coupling, as well as the technique advantages and performance characteristics. A large compilation of CE-MS applications is also presented. Therefore, this review is both a guide for beginners and a collection of key references for people who are familiar to the technique. Furthermore, this is the first CE-MS review published in a Brazilian journal and marks the installation of the first two commercial CE-MS units in Sao Paulo State.

Estratégias de pré-concentração em eletroforese capilar (CE): parte 1. Manipulação da velocidade eletroforética do analito

Capillary electrophoresis has become a well-established and routine-based separation technique. It is based on the differences between charged analyte mobility in aqueous or organic electrolytes. Its major limitation is the sensitivity due to small sample injection volumes and the narrow diameter of the capillaries, especially when UV detection is used. There are a number of ways to increase the concentration sensitivity. This report shows some on-line preconcentration strategies to perform it in free solution capillary electrophoresis that are based on manipulation of the analyte electrophoretic velocity during the sample introduction (stacking, field amplification and transient isotachophoresis).

Níveis de lisina digestível para frangos de corte machos no período de 37 a 49 dias de idade

Foram utilizados 1.015 frangos de corte machos, linhagem Ross, dos 37 aos 49 dias de idade, para avaliar os efeitos de diferentes níveis de lisina digestível nas dietas experimentais. O delineamento experimental foi inteiramente casualizado, com 5 níveis de lisina digestível (0,90; 0,95; 1,00; 1,05; 1,10%), cada um com 7 repetições e 29 aves por unidade experimental. Utilizaram-se dietas isoenergéticas com 3.250 de kcal de EM/kg e isoprotéicas, com 18% de PB, à base de milho e farelo de soja. Foram avaliados o ganho de peso, o consumo de ração, a conversão alimentar, as características de carcaça, o rendimento de cortes, a composição e a deposição de nutrientes corporais. Os níveis de lisina digestível influenciaram, entre as características de desempenho, apenas a conversão alimentar, que melhorou de forma linear de acordo com níveis de lisina da ração. Das características de carcaça e rendimento de cortes, apenas gordura abdominal aumentou de forma quadrática conforme os níveis de lisina. Os níveis de lisina digestível tiveram efeito quadrático no teor da matéria mineral da carcaça, mas não influenciaram a composição química das vísceras e do sangue. Observou-se, contudo, tendência a aumento linear na deposição de proteína da carcaça e do corpo vazio com o aumento no nível de lisina digestível. Os resultados de desempenho indicam que o nível de lisina digestível da ração de frangos de corte machos no período de 37 a 49 dias de idade deve ser igual ou superior a 1,10%.

Fosfatos com diferentes relações flúor: fósforo na alimentação de frangos de corte

Um experimento foi conduzido com o objetivo de verificar os efeitos de diferentes relações flúor:fosforo na alimentação sobre o desempenho de frangos de corte. Foram utilizados 1.000 pintos de corte de 1 dia distribuídos em um delineamento inteiramente casualizado, com quatro tratamentos e cinco repetições de 50 aves por boxe. Os tratamentos consistiram de quatro fontes de fósforo com relações flúor:fósforo de 1:40, 1:60, 1:80 e 1:100. O experimento foi dividido em três fases experimentais: 1 a 21, 22 a 42 e 43 a 49 dias de idade. Em cada fase, avaliou-se o consumo de ração, o ganho de peso e a conversão alimentar. Ao final do experimento, foram abatidas duas aves de cada repetição para coleta da tíbia e de músculos do peito para análise das concentrações de flúor, cálcio, fósforo e magnésio desses tecidos. As relações flúor:fósforo avaliadas não afetaram o desempenho dos frangos de corte. A deposição óssea de flúor é proporcional à sua concentração na dieta, no entanto, a elevação dos níveis de flúor da dieta não influencia sua deposição nos músculos.

Dietary levels of lysine for male broilers from 23 to 36 days of age: performance and body composition

One thousand and fifty (1,050) Ross male broilers from 23 to 36 days of age were used to evaluate the dietary effects of levels of lysine. It was used a random block design with 5 levels of digestible lysine (0.95; 1.00; 1.05; 1.10 and 1.15%). Each level was applied to seven replicates and the experimental unit was composed of 30 birds. Diets were offered in seven replicates and the experimental unit was represented by 30 birds. Weight gain, feed intake, feed conversion, carcass traits, cut yields, composition and deposition of body nutrients were evaluated. The concentration of 1.09% of digestible lysine optimized performance without significant effects on feed intake. Carcass traits and cut yields had no significant differences among levels of lysine. On the other hand, there was a quadratic effect on carcass weight, which was the best at the level of 1.07% of digestible lysine. On crude protein content of the empty body, the best response was observed at the level of 1.10% of dietary lysine. This same level corresponded to a quadratic response in the deposition of water from carcass and empty body. The level of digestible lysine for better performance, chemical composition and body nutrient deposition in broilers from 23 to 36 days of age is between 1.07 and 1.10%.

Fontes de minerais orgânicos e inorgânicos para leitões desmamados

A presente pesquisa foi realizada com o objetivo de comparar os efeitos de fontes orgânicas e inorgânicas de microminerais sobre o desempenho, os parâmetros sanguíneos e a deposição de minerais em tecidos e órgãos de leitões desmamados. Foram utilizados 54 leitões de genética comercial, desmamados com idade média de 24 dias e peso médio de 7,35±0,88kg, num delineamento experimental de blocos ao acaso, com dois tratamentos, nove repetições e três animais por parcela. Os tratamentos foram representados pelo uso de rações suplementadas com minerais de fontes orgânicas ou inorgânicas (Cu, Zn, Fe, Mn e Se) no período dos 24 aos 57 dias de idade. Foram avaliados o consumo diário de ração, o ganho diário de peso, a conversão alimentar, os parâmetros hematológicos e a deposição de minerais em tecidos e órgãos. As fontes orgânicas de minerais nas rações fornecidas dos 24 aos 57 dias de idade melhoraram o ganho diário de peso (P=0,06) e a conversão alimentar (P=0,05) e aumentaram o número de hemáceas (P=0,10), contudo não influenciaram a deposição de minerais no músculo masseter, fígado, coração, baço e rim. Portanto, o suplemento de minerais de fontes orgânicas foi mais eficiente para o desempenho dos leitões na fase de creche.

Lisina digestível para frangos de corte machos entre 12 e 22 dias de idade

Avaliou-se nível de lisina digestível para 1050 frangos de corte dos 12 aos 22 dias de idade. O delineamento experimental foi inteiramente ao acaso, com cinco tratamentos, sete repetições e 30 aves por unidade experimental. Os tratamentos foram: 1,05; 1,10; 1,15; 1,20 e 1,25% de lisina digestível. Avaliaram-se ganho de peso, consumo de ração, conversão alimentar, composição e deposição de nutrientes corporais. Foram constatados efeitos quadráticos de lisina digestível no consumo de ração e resposta linear ascendente no peso da carcaça. Na composição química da carcaça, houve resposta quadrática do nível de lisina na concentração de proteína. As taxas de deposição proteica, deposição de água, da carcaça e do corpo total tiveram aumento linear em resposta ao acréscimo de lisina na dieta. O aumento da concentração de lisina, todavia, coincidiu com a redução da matéria mineral nas vísceras e sangue e no corpo total. Considerado o desempenho, o nível 1,1% de lisina digestível atendeu às necessidades do frango de corte entre o 12º e o 22º dia de idade. Consideradas a composição química e as taxas de deposição dos nutrientes corporais, a demanda pelo aminoácido digestível torna-se igual ou maior que 1,25%

Digestible lysine and organic zinc for male broiler from 1 to 11 days of age: performance and body composition

This study evaluated levels of digestible lysine and organic zinc for male Ross strain broilers from 1 to 11 days of age. It was used 1,050 chicks distributed in randomized block design, in 5 × 2 factorial scheme, with seven repetitions of 15 birds per experimental unit. The dietary concentrations of digestible lysine were 0.90; 1.00; 1.10; 1.20; and 1.40% combined with 43 and 253 ppm zinc chelate. The diets contained 2,965 ± 18 kcal/kg of apparent metabolizable energy (AME) and 21.48 ± 0.18% of CP. It was determined chemical composition, protein, lipid, mineral and water depositions on carcass and empty body. There was no interaction among the factors digestible lysine and organic zinc. Effect of zinc concentration increase was observed on greatest deposition of body fat, indicating that there is interference in lipid metabolism of the birds in the studied phase. The most pronounced effects resulted from the dietary inclusion of lysine. There was a linear effect on reconstituted body weight as a response to the increase of dietary lysine, which suggests equal or superior requirement to the greatest studied level. However, the deposition of water and protein on the carcass had a quadratic increase, characterizing higher muscle mass accumulation up to the levels 1.25 and 1.27 of this amino acid in the diet. Considering the studied strain, broiler chickens from the first to the 11th days of age require 1.28 ± 0.01% of digestible lysine, according to the deposition of muscle mass.

Lysine and zinc chelate in diets for brown laying hens: effects on egg production and composition

It was evaluated the effects of levels of digestible lysine and chelate zinc combined in the diet for laying on the egg quality. It was used 720 birds, from 48 to 60 weeks of age, distributed in a completely randomized design in a 3 × 5 factorial scheme with three levels of zinc and five levels of lysine, applied into six replicates in the experimental units of eight birds per plot. The levels were: 137, 309 and 655 ppm zinc and 0.482, 0.527, 0.582, 0.644 and 0.732% digestible lysine. It was not observed any interaction among digestible lysine and zinc for the primary variables of fractions and egg composition. Levels of zinc reduced egg weight, suggesting the lowest efficiency in nutrient intake. At the highest dietary concentration of zinc, the addition of digestible lysine coincided with a linear increase in shell weight. However, zinc addition, regardless of lysine level in the diet, resulted in the reduction of egg weight and of the percentage of mineral matter in the yolk, limiting the efficiency of mineral deposition in this fraction of the egg. Concentration of zinc that produced the best results was 137 ppm inasmuch as higher quantities limit the use of digestible lysine, with effects harming composition and egg quality. The study indicates the following requirement for digestible lysine: 0.639% from the 48th to the 52nd week, 0.679% from the 52nd to 56th week, and 0.635% from the 56th to the 60th week. Considering the total period from 48th to the 60th week, the level 0.638% of lysine or the daily intake of 707 mg of the amino acid met the requirement for egg quality of semi-heavy layers used in this study.

Lisina digestível e zinco orgânico para frangos de corte machos na fase de 22 a 42 dias de idade

Os efeitos dietéticos de lisina digestível e zinco quelato para frangos de corte machos entre 22 e 42 dias de idade foram avaliados em dois ensaios experimentais. Adotou-se o delineamento inteiramente casualizado em arranjo fatorial 5 × 2, com cinco níveis de lisina (0,841; 0,876; 0,997; 1,022 e 1,030%) e dois de zinco (43 e 243 ppm). No primeiro ensaio, utilizaram-se 900 aves com peso médio inicial de 957,4 g distribuídas em parcelas experimentais de 30 aves e três repetições e, no segundo ensaio, 180 frangos com peso médio inicial de 866,22 g, divididos em parcelas de três aves e seis repetições. As características avaliadas foram desempenho, rendimento de cortes e composição corporal (1º ensaio), balanço de nitrogênio e digestibilidade aparente das dietas (2º ensaio). Houve interação entre os níveis de lisina e zinco para o ganho de peso, a conversão alimentar, a matéria seca ingerida e o balanço energético. Os rendimentos de peito, coxa e sobrecoxa tiveram aumentos lineares em resposta ao acréscimo no nível de lisina digestível na dieta. O melhor desempenho foi obtido com o nível de 0,997% de lisina digestível (ou 1,134% lisina total) e de 43 ppm de zinco. Para maior rendimento dos cortes, o nível de lisina digestível recomendável deve ser no mínimo 1,002% (ou 1,139% de lisina total), independentemente dos níveis de zinco quelato. A maior inclusão de zinco em dietas para frangos de corte dos 22 aos 42 dias de idade não melhora a utilização da lisina na dieta e aumenta a deposição de gordura corporal.

Study of metal oxide-semiconductor capacitors with rf magnetron sputtering TiO(x) and TiO(x)N(y) gate dielectric layer

Metal oxide-semiconductor capacitors with TiO(x) deposited with different O(2) partial pressures (30%, 35%, and 40%) and annealed at 550, 750, and 1000 degrees C were fabricated and characterized. Fourier transform infrared, x-ray near edge spectroscopy, and elipsometry measurements were performed to characterize the TiO(x) films. TiO(x)N(y) films were also obtained by adding nitrogen to the gaseous mixture and physical results were presented. Capacitance-voltage (1 MHz) and current-voltage measurements were utilized to obtain the effective dielectric constant, effective oxide thickness, leakage current density, and interface quality. The results show that the obtained TiO(x) films present a dielectric constant varying from 40 to 170 and a leakage current density (for V(G)=-1 V, for some structures as low as 1 nA/cm(2), acceptable for complementary metal oxide semiconductor circuits fabrication), indicating that this material is a viable, in terms of leakage current density, highk substitute for current ultrathin dielectric layers. (C) 2009 American Vacuum Society. [DOI: 10.1116/1.3043537]

Changes in integument structure during the imaginal molt of the honey bee

The changing pattern of developing cuticle and associated epidermis is described during the imaginal molt in the honey bee. Observations began immediately after the pupal molt, and included histological analyses of the integument during apolysis and the subsequent deposition and differentiation of the adult cuticle. Apolysis coincides with a marked increase in the thickness and reorganization of the epidermal layer, reflecting changes in cell structure. The epidermis remains thickened during the period of cuticle deposition, suggesting intense biosynthetic activity, but turns into a very thin layer during cuticle differentiation, clearly indicating that secretory activity for cuticle formation is terminating. The thoracic cuticle differentiates earlier and becomes thicker than the abdominal. The observed changes in integument structure provide insights that permit an improved physiological characterization for staging pupal and pharate adult development.

Ecdysteroid-Dependent Expression of the Tweedle and Peroxidase Genes during Adult Cuticle Formation in the Honey Bee, Apis mellifera

Cuticle renewal is a complex biological process that depends on the cross talk between hormone levels and gene expression. This study characterized the expression of two genes encoding cuticle proteins sharing the four conserved amino acid blocks of the Tweedle family, AmelTwdl1 and AmelTwdl2, and a gene encoding a cuticle peroxidase containing the Animal haem peroxidase domain, Ampxd, in the honey bee. Gene sequencing and annotation validated the formerly predicted tweedle genes, and revealed a novel gene, Ampxd, in the honey bee genome. Expression of these genes was studied in the context of the ecdysteroid-coordinated pupal-to-adult molt, and in different tissues. Higher transcript levels were detected in the integument after the ecdysteroid peak that induces apolysis, coinciding with the synthesis and deposition of the adult exoskeleton and its early differentiation. The effect of this hormone was confirmed in vivo by tying a ligature between the thorax and abdomen of early pupae to prevent the abdominal integument from coming in contact with ecdysteroids released from the prothoracic gland. This procedure impaired the natural increase in transcript levels in the abdominal integument. Both tweedle genes were expressed at higher levels in the empty gut than in the thoracic integument and trachea of pharate adults. In contrast, Ampxd transcripts were found in higher levels in the thoracic integument and trachea than in the gut. Together, the data strongly suggest that these three genes play roles in ecdysteroid-dependent exoskeleton construction and differentiation and also point to a possible role for the two tweedle genes in the formation of the cuticle (peritrophic membrane) that internally lines the gut.

Loxosceles gaucho Venom-Induced Acute Kidney Injury - In Vivo and In Vitro Studies

Background: Accidents caused by Loxosceles spider may cause severe systemic reactions, including acute kidney injury (AKI). There are few experimental studies assessing Loxosceles venom effects on kidney function in vivo. Methodology/Principal Findings: In order to test Loxosceles gaucho venom (LV) nephrotoxicity and to assess some of the possible mechanisms of renal injury, rats were studied up to 60 minutes after LV 0.24 mg/kg or saline IV injection (control). LV caused a sharp and significant drop in glomerular filtration rate, renal blood flow and urinary output and increased renal vascular resistance, without changing blood pressure. Venom infusion increased significantly serum creatine kinase and aspartate aminotransferase. In the LV group renal histology analysis found acute epithelial tubular cells degenerative changes, presence of cell debris and detached epithelial cells in tubular lumen without glomerular or vascular changes. Immunohistochemistry disclosed renal deposition of myoglobin and hemoglobin. LV did not cause injury to a suspension of fresh proximal tubules isolated from rats. Conclusions/Significance: Loxosceles gaucho venom injection caused early AKI, which occurred without blood pressure variation. Changes in glomerular function occurred likely due to renal vasoconstriction and rhabdomyolysis. Direct nephrotoxicity could not be demonstrated in vitro. The development of a consistent model of Loxosceles venom-induced AKI and a better understanding of the mechanisms involved in the renal injury may allow more efficient ways to prevent or attenuate the systemic injury after Loxosceles bite.

Collagen V-induced nasal tolerance downregulates pulmonary collagen mRNA gene and TGF-beta expression in experimental systemic sclerosis

Background: The purpose of this study was to evaluate collagen deposition, mRNA collagen synthesis and TGFbeta expression in the lung tissue in an experimental model of scleroderma after collagen V-induced nasal tolerance. Methods: Female New Zealand rabbits (N = 12) were immunized with 1 mg/ml of collagen V in Freund's adjuvant (IM). After 150 days, six immunized animals were tolerated by nasal administration of collagen V ( 25 mu g/day) (IM-TOL) daily for 60 days. The collagen content was determined by morphometry, and mRNA expressions of types I, III and V collagen were determined by Real-time PCR. The TGF-beta expression was evaluated by immunostaining and quantified by point counting methods. To statistic analysis ANOVA with Bonferroni test were employed for multiple comparison when appropriate and the level of significance was determined to be p < 0.05. Results: IM-TOL, when compared to IM, showed significant reduction in total collagen content around the vessels (0.371 +/- 0.118 vs. 0.874 +/- 0.282, p < 0.001), bronchioles (0.294 +/- 0.139 vs. 0.646 +/- 0.172, p < 0.001) and in the septal interstitium (0.027 +/- 0.014 vs. 0.067 +/- 0.039, p = 0.026). The lung tissue of IM-TOL, when compared to IM, showed decreased immunostaining of types I, III and V collagen, reduced mRNA expression of types I (0.10 +/- 0.07 vs. 1.0 +/- 0.528, p = 0.002) and V (1.12 +/- 0.42 vs. 4.74 +/- 2.25, p = 0.009) collagen, in addition to decreased TGF-beta expression ( p < 0.0001). Conclusions: Collagen V-induced nasal tolerance in the experimental model of SSc regulated the pulmonary remodeling process, inhibiting collagen deposition and collagen I and V mRNA synthesis. Additionally, it decreased TGF-beta expression, suggesting a promising therapeutic option for scleroderma treatment.