Efeitos da relação volumoso: concentrado e do peso ao abate sobre os componentes da perna de cordeiros Ile de France x Ideal confinados

Foram avaliados os seguintes componentes da perna dos animais: peso total e comprimento, peso total de músculos, peso dos músculos bíceps femural + semimembranoso + semitendinoso + quadríceps femural + adutor, peso total de ossos, peso do fêmur e das gorduras subcutânea, intermuscular e total, porcentagens de todos estes pesos em relação ao peso total da perna, comprimento e circunferência do fêmur e calculados a relação músculo:osso e o índice de musculosidade. Foram utilizados 20 cordeiros inteiros Ile de France x Ideal (F1) com peso vivo médio ao início do experimento de 18,2 ± 0,74 kg e idade média de 83 ± 10 dias. As relações volumoso (V):concentrado (C) utilizadas no experimento foram 50V:50C ou 30V:70C, com base na matéria seca (MS), sendo as rações isoprotéicas (18% de proteína bruta na MS) e isoenergéticas (10,92 MJ de energia metabolizável/kg MS). Os animais foram abatidos aos 30 ou 34 kg de peso vivo. Os abatidos mais pesados apresentaram perna, total de músculos, músculos bíceps femural + semimembranoso + semitendinoso + quadríceps femural + adutor, total de ossos, fêmur e gorduras subcutânea e total mais pesados, além de maior circunferência do fêmur. Os animais que receberam a ração com relação 30V:70C apresentaram menores relação músculo:osso e índice de musculosidade e maior porcentagem de gordura intermuscular em relação ao peso total da perna.

Protein levels and environmental temperature effects on carcass characteristics, performance, and nitrogen excretion of broiler chickens from 7 to 21 days of age

This trial was conducted to evaluate the utilization of low-protein diets formulated based on the ideal protein concept for broiler chickens from 7 to 21 days of age reared at different environmental temperatures. Nine hundred male Cobb-500® chickens were used. At day seven chicks were distributed according to a completely randomized design in a 3 x 3 factorial with four replications of 25 birds each. It was used three crude protein levels in the diet (21.5; 20.0 and 18.5%) and three environmental temperatures (low, thermoneutral and high). The performance, carcass characteristics (yield and chemical composition), and nitrogen ingestion and excretion were assessed. There was no significant interaction among the factors for the evaluated variables. Environmental temperatures affected differently chicken performance. High environmental temperature resulted in lower weight gain and higher wing fat percentage, whereas cold temperature resulted in higher feed conversion. on the other hand, low-protein diets decreased weight gain, breast yield, nitrogen excretion and influenced breast and wings chemical composition. Birds reared at high environmental temperature showed lower nitrogen intake and excretion. The results showed that the decrease in protein levels from 7 to 21 days of age contributed to lower nitrogen excretion in broiler chickens, but impaired performance and carcass characteristics independent of rearing temperature.

Development and application of a Saccharomyces cerevisiae-expressed nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibodies against infectious bronchitis virus

A Saccharomyces cerevisiae-expressed nucleocapsid (N) polypeptide of the M41 strain of infectious bronchitis virus (IBV) was used as antigen in a recombinant yeast-expressed N protein-based enzyme-linked immunosorbent assay (Y-N-ELISA). The Y-N-ELISA was rapid, sensitive, and specific for detecting chicken serum antibodies to IBV, and it compared favorably with a commercial ELISA.

Características físico-qímicas da carne de caprinos submetidos a diferentes níveis de substituição do leite por soro de queijo durante o aleitamento

Fifty six crossbred kids Parda Alpina x Gurgueia were separeted into four groups of 14 (seven male and seven female) and submited to four treatments with different levels of milk goat substitution by cheese serum. The treatments were : T1=milk goat (testify); T2=15% serum at 23 days and 33% at 55 days; T3=15% serum starting from 16 days, 33% on 23 days and 51% on 72 days of age; and T4=33% on 16 days and 69% of liquid diet in the final fase. At 92 days of age, all animals were slaughtered for carcass evaluation. Pallete were separeted from 20 male animals (five by treatment) and the physical and chemical analysis of the meat were realized using these materials. The studied characteristics were: total weight (TW), meat weight (MW), bone weigth (BW) of the pallete, and percentage of umidity (U), ashes (A), protein (P), fat (F) and pH of the pallete meat. There was significant effect of the treatment over F. Means of PW, A and BW were, respectively 491, 342 and 149.8 g. The meal represented 69,6% of total pallete weight. Means of U, A, P and F were 73, 1.0, 19 and 1.95%, respectively. The pH was 6.7. Results allow to conclude that only the fat percentage of the meat was modified, due to the administration of serun milk on the liquid diet of the kids, during the milking fase.

Culture of osteogenic cells from human alveolar bone: A useful source of alkaline phosphatase

The aim of this study was to obtain membrane-bound alkaline phosphatase from osteoblastic-like cells of human alveolar bone. Cells were obtained by enzymatic digestion and maintained in primary culture in osteogenic medium until subconfluence. First passage cells were cultured in the same medium and at 7, 14, and 21 days, total protein content, collagen content, and alkaline phosphatase activity were evaluated. Bone-like nodule formation was evaluated at 21 days. Cells in primary culture at day 14 were washed with Tris-HCl buffer, and used to extract the membrane-bound alkaline phosphatase. Cells expressed osteoblastic phenotype. The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10.0. This enzyme also hydrolyzes ATP, ADP, fructose-1-phosphate, fructose-6-phosphate, pyrophosphate and beta-glycerophosphate. PNPPase activity was reduced by typical inhibitors of alkaline phosphatase. SDS-PAGE of membrane fraction showed a single band with activity of similar to 120 kDa that could be solubilized by phospholipase C or Polidocanol. (c) 2007 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Membrane-bound alkaline phosphatase from ectopic mineralization and rat bone marrow cell culture

Cells from rat bone marrow exhibit the proliferation-differentiation sequence of osteoblasts, form mineralized extracellular matrix in vitro and release alkaline phosphatase into the medium. Membrane-bound alkaline phosphatase was obtained by method that is easy to reproduce, simpler and fast when compared with the method used to obtain the enzyme from rat osseous plate. The membrane-bound alkaline phosphatase from cultures of rat bone marrow cells has a MWr of about 120 kDa and specific PNPP activity of 1200 U/tng. The ecto-enzyme is anchored to the plasma membrane by the GPI anchor and can be released by PIPLC (selective treatment) or polidocanol (0.2 mg/mL protein and 1% (w/v) detergent). The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10. This fraction hydrolyzes ATP (240 U/mg), ADP (350 U/ mg), glucose 1-phosphate (1100 U/mg), glucose 6-phosphate (340 Wing), fructose 6-phosphate (460 U/mg), pyrophosphate (330 U/mg) and (3glycerophosphate (600 U/mg). Cooperative effects were observed for the hydrolysis of PPi and beta-glycerophosphate. PNPPase activity was inhibited by 0.1 mM vanadate (46%), 0.1 mM ZnCl2 (68%), 1 mM levamisole (66%), 1 mM arsenate (44%), 10 mM phosphate (21%) and 1 mM theophylline (72%). We report the biochemical characterization of membrane-bound alkaline phosphatase obtained from rat bone marrow cells cultures, using a method that is simple, rapid and easy to reproduce. Its properties are compared with those of rat osseous plate enzyme and revealed that the alkaline phosphatase obtained has some kinetics and structural behaviors with higher levels of enzymatic activity, facilitating the comprehension of the mineralization process and its function. (c) 2006 Elsevier B.V. All rights reserved.

Effects of energy and protein levels on egg quality and performance of laying hens at early second production cycle

One hundred sixty-two commercial 70-wk-old ISA Brown laying hens, previously subjected to induced molting by feed restriction, were distributed in a completely randomized design with 3 x 3 factorial arrangement (i.e., 3 metabolizable energy levels: 2,850; 2,950, and 3,050 kcal of ME/kg) and 3 protein levels (16, 18, and 20% CP), which totaled 9 treatments with 3 replicates of 6 birds each. Experimental diets were offered to birds after the feed restriction period. Performance and egg quality parameters were evaluated in 14-d intervals from the 4th to 12th weeks after forced molting for a total of 4 evaluation periods. Increases in dietary energy and protein levels did not improve performance or egg quality. The levels of 2,850 kcal of ME and 16% protein were sufficient for laying hens starting the second production cycle without decreasing their performance or egg quality.

Genetic parameters for milk, fat and protein yields in Murrah buffaloes (Bubalus bubalis Artiodactyla, Bovidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estimativa da composição química corporal de tourinhos Santa Gertrudes a partir da composição química e física das 9-10-11ª costelas

Foram obtidas equações de regressão linear simples para estimar a composição química corporal de bovinos Santa Gertrudes, a partir da composição química e física do corte das 9-10-11ª costelas. Quinze tourinhos, entre nove a 15 meses de idade e de 220 a 505 kg de peso, foram mantidos confinados. Os animais foram abatidos após jejum completo de 18 horas, sendo que seis deles foram abatidos após adaptação. A composição química em água, proteína, extrato etéreo e minerais foi determinada no corte das costelas e em amostras obtidas após moagem completa e homogeneização de todos os tecidos corporais, divididos em: sangue, couro, cabeça + patas, vísceras e carcaça. A composição física do corte das costelas foi obtida por separação manual do músculo, gordura e ossos. O peso do corpo vazio foi altamente correlacionado ao peso da carcaça quente (r² = 0,99). As porcentagens de água e extrato etéreo das 9-10-11ª costelas mostraram-se altamente correlacionadas com a composição química do corpo vazio, o que não ocorreu para as porcentagens de proteína e minerais. Esses teores foram calculados pela composição do corpo vazio desengordurado. A composição física do corte das costelas foi eficiente para estimar as porcentagens de água, extrato etéreo e minerais do corpo vazio, utilizando-se a porcentagem de gordura separável das costelas, mas não para estimar o teor de proteína. A composição física do corte das costelas demonstrou ser uma técnica eficiente, mas a composição química apresentou maiores coeficientes de determinação e menores erros da estimativa. Como a porcentagem de água no corpo vazio e no corte das costelas (r² = 0,95), e as porcentagens de água e de extrato etéreo no corpo vazio foram altamente correlacionadas (r² = 0,94), a porcentagem de água no corte das 9-10-11ª costelas poderia ser a única variável para estimativa da composição química corporal.

Prediction equations to estimate the demand of energy and crude protein for maintenance, gain and egg production for laying Japanese quails

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Latex use as an occlusive membrane for guided bone regeneration

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

Splice variants of the human ZC3H14 gene generate multiple isoforms of a zinc finger polyadenosine RNA binding protein

The human ZC3H14 gene encodes an evolutionarily conserved Cys(3)His zinc finger protein that binds specifically to polyadenosine RNA and is thus postulated to modulate post-transcriptional gene expression. Expressed sequence tag (EST) data predicts multiple splice variants of both human and mouse ZC3H14. Analysis of ZC3H14 expression in both human cell lines and mouse tissues confirms the presence of multiple alternatively spliced transcripts. Although all of these transcripts encode protein isoforms that contain the conserved C-terminal zinc finger domain, suggesting that they could all bind to polyadenosine RNA, they differ in other functionally important domains. Most of the alternative transcripts encode closely related proteins (termed isoforms 1, 2. 3, and 3short) that differ primarily in the inclusion of three small exons, 9, 10, and 11, resulting in predicted protein isoforms ranging from 82 to 64 kDa. Each of these closely related isoforms contains predicted classical nuclear localization signals (cNLS) within exons 7 and 11. Consistent with the presence of these putative nuclear targeting signals, these ZC3H14 isoforms are all localized to the nucleus. In contrast, an additional transcript encodes a smaller protein (34 kDa) with an alternative first exon (isoform, 4). Consistent with the absence of the predicted cNLS motifs located in exons 7 and 11, ZC3H14 isoform 4 is localized to the cytoplasm. Both EST data and experimental data suggest that this variant is enriched in testes and brain. Using an antibody that detects endogenous ZC3H14 isoforms 1-3 reveals localization of these isoforms to nuclear speckles. These speckles co-localize with the splicing factor, SC35, suggesting a role for nuclear ZC3H14 in mRNA processing. Taken together, these results demonstrate that multiple transcripts encoding several ZC3H14 isoforms exist in vivo. Both nuclear and cytoplasmic ZC3H14 isoforms could have distinct effects on gene expression mediated by the common Cys(3)His zinc finger polyadenosine RNA binding domain. (C) 2009 Elsevier B.V. All rights reserved.

Loss of nuclear poly(A)-binding protein 1 causes defects in myogenesis and mRNA biogenesis

The nuclear poly(A)-binding protein 1 (PABPN1) is a ubiquitously expressed protein that plays a critical role in polyadenylation. Short expansions of the polyalanine tract in the N-terminus of PABPN1 lead to oculopharyngeal muscular dystrophy (OPMD), which is an adult onset disease characterized by eyelid drooping, difficulty in swallowing and weakness in the proximal limb muscles. Although significant data from in vitro biochemical assays define the function of PABPN1 in control of poly(A) tail length, little is known about the role of PABPN1 in mammalian cells. To assess the function of PABPN1 in mammalian cells and specifically in cells affected in OPMD, we examined the effects of PABPN1 depletion using siRNA in primary mouse myoblasts from extraocular, pharyngeal and limb muscles. PABPN1 knockdown significantly decreased cell proliferation and myoblast differentiation during myogenesis in vitro. At the molecular level, PABPN1 depletion in myoblasts led to a shortening of mRNA poly(A) tails, demonstrating the cellular function of PABPN1 in polyadenylation control in a mammalian cell. In addition, PABPN1 depletion caused nuclear accumulation of poly(A) RNA, revealing that PABPN1 is required for proper poly(A) RNA export from the nucleus. Together, these experiments demonstrate that PABPN1 plays an essential role in myoblast proliferation and differentiation, suggesting that it is required for muscle regeneration and maintenance in vivo.

Investigation of the interaction between Tc85-11 protein and antibody anti-T cruzi by AFM and amperometric measurements

This present work reports on development of an amperometric immunosensor for the diagnosis of Chagas' disease using a specific glycoprotein of the trypomastigote surface, which belongs to the Tc85-11 protein family of Trypanosoma cruzi (T cruzi). An atomically flat gold surface on a silicon substrate and gold screen-printed electrodes were functionalized with cystatrine and later activated with glutaraldehyde (GA), which was used to form covalent bonds with the purified recombinant antigen (Tc85-11). The antigen reacts with the antibody from the serum, and the affinity reaction was monitored directly using atomic force microscopy or amperometry through a secondary antibody tagged to peroxidase (HRP). Surface imaging allowed to us to differentiate the modification steps and antigen-antibody interaction allowed to distinguish the affinity reactions. In the amperometric immunosensor, peroxidase catalyses the L-2 formation in the presence of hydrogen peroxide and potassium iodide, and the reduction current intensity was measured at a given potential with screen-printed electrodes. The immunosensor was applied to sera of chagasic patients and patients having different systemic diseases. (c) 2006 Elsevier Ltd. All rights reserved.

Natural rubber latex used as drug delivery system in guided bone regeneration (GBR)

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