CalDAG-GEFI is at the nexus of calcium-dependent platelet activation.

The importance of the second messengers calcium (Ca(2+)) and diacylglycerol (DAG) in platelet signal transduction was established more than 30 years ago. Whereas protein kinase C (PKC) family members were discovered as the targets of DAG, little is known about the molecular identity of the main Ca(2+) sensor(s). We here identify Ca(2+) and DAG-regulated guanine nucleotide exchange factor I (CalDAG-GEFI) as a critical molecule in Ca(2+)-dependent platelet activation. CalDAG-GEFI, through activation of the small GTPase Rap1, directly triggers integrin activation and extracellular signal-regulated kinase-dependent thromboxane A(2) (TxA(2)) release. CalDAG-GEFI-dependent TxA(2) generation provides crucial feedback for PKC activation and granule release, particularly at threshold agonist concentrations. PKC/P2Y12 signaling in turn mediates a second wave of Rap1 activation, necessary for sustained platelet activation and thrombus stabilization. Our results lead to a revised model for platelet activation that establishes one molecule, CalDAG-GEFI, at the nexus of Ca(2+)-induced integrin activation, TxA(2) generation, and granule release. The preferential activation of CalDAG-GEFI over PKC downstream of phospholipase C activation, and the different kinetics of CalDAG-GEFI- and PKC/P2Y12-mediated Rap1 activation demonstrate an unexpected complexity to the platelet activation process, and they challenge the current model that DAG/PKC-dependent signaling events are crucial for the initiation of platelet adhesion.

Effect of arginine on the development of the pectoralis muscle and the diameter and the protein:deoxyribonucleic acid rate of its skeletal myofibers in broilers

This work aimed at evaluating the effects of the supplementation of starter diet with Arg on breast muscle development in broilers and the activation of satellite cells and the aggregation of myofibrillar protein. Male Cobb chicks (n = 990) were randomly assigned to 1 of 5 treatments in a complete random design. Measurements of 33 chicks per treatment were made in 6 repetitions. The treatments consisted of a basal diet with 1.390% digestible Arg (without supplementation) and 4 dietary levels of Arg (1.490, 1.590, 1.690, and 1.790%) with Arg:Lys ratios of 1.103, 1.183, 1.262, 1.341, and 1.421, respectively. Arginine supplementation was used only in the starter phase (1 to 21 d). Dietary supplementation with Arg had a positive effect (P < 0.05) on breast and breast fillet weight on d 7 and 21 and on myofiber diameter on d 14 and 21. However, no effect was observed (P > 0.05) on the protein: DNA ratio, which demonstrates that Arg does not interfere with the mitotic activity of the satellite cells. Independently from mechanism, Arg affected muscle growth in the starter phase positively. Dietary supplementation with Arg in the starter phase had no effect (P > 0.05) on the carcass yield of broilers on d 42. Diet supplementation with Arg at levels above the ones recommended for the starter phase may be necessary for improved muscle development in broilers.

Involvement of Phosphatidylinositol-3 Kinase/AKT/PKC zeta/lambda Pathway in the Effect of Palmitate on Glucose-Induced Insulin Secretion

Objectives: In the present study, a novel pathway by which palmilate potentiates glucose-induced insulin secretion by pancreatic beta cells was investigated. Methods: Groups of freshly isolated islets were incubated in 10 mM glucose with palmitate, LY294002, wortmannin, and fumonism B I for measurement of insulin secretion by radioimmunoassay (RIA). Also, phosphorylation and content of AKT and PKC proteins were evaluated by immunoblotting. Results: Glucose plus palmitate and glucose plus LY294002 or wortmannin (PI3K inhibitors) increased glucose-induced insulin secretion by isolated pancreatic islets. Glucose at 10 mM induced AKT and PKC zeta/lambda phosphorylation. Palmitate (0.1 mM) abolished glucose stimulation of AKT and PKC zeta/lambda phosphorylation possibly through PI3K inhibition because both LY294002 (50 mu M) and wortmannin (100 nM) caused the same effect. The inhibitory effect of palmitate on glucose-induced AKT and PKC zeta/lambda phosphorylation and the stimulatory effect of palmitate on glucose-induced insulin secretion were not observed in the presence of fumonisin B1, all inhibitor of ceramide synthesis. Conclusions: These findings support the proposition that palmilate increases insulin release in the presence of 10 mM glucose by inhibiting PI3K activity through a mechanism that involves ceramide synthesis.

Activation of insulin and IGF-1 signaling pathways by melatonin through MT1 receptor in isolated rat pancreatic islets

Melatonin diminishes insulin release through the activation of MT1 receptors and a reduction in cAMP production in isolated pancreatic islets of neonate and adult rats and in INS-1 cells ( an insulin-secreting cell line). The pancreas of pinealectomized rats exhibits degenerative pathological changes with low islet density, indicating that melatonin plays a role to ensure the functioning of pancreatic beta cells. By using immunoprecipitation and immunoblotting analysis we demonstrated, in isolated rat pancreatic islets, that melatonin induces insulin growth factor receptor (IGF-R) and insulin receptor (IR) tyrosine phosphorylation and mediates the activities of the PI3K/AKT and MEK/ERKs pathways, which are involved in cell survival and growth, respectively. Thus, the effects of melatonin on pancreatic islets do not involve a reduction in cAMP levels only. This indoleamine may regulate growth and differentiation of pancreatic islets by activating IGF-I and insulin receptor signaling pathways.

Functional Characterization of LcpA, a Surface-Exposed Protein of Leptospira spp. That Binds the Human Complement Regulator C4BP

We have previously shown that pathogenic leptospiral strains are able to bind C4b binding protein (C4BP). Surface-bound C4BP retains its cofactor activity, indicating that acquisition of this complement regulator may contribute to leptospiral serum resistance. In the present study, the abilities of seven recombinant putative leptospiral outer membrane proteins to interact with C4BP were evaluated. The protein encoded by LIC11947 interacted with this human complement regulator in a dose-dependent manner. The cofactor activity of C4BP bound to immobilized recombinant LIC11947 (rLIC11947) was confirmed by detecting factor I-mediated cleavage of C4b. rLIC11947 was therefore named LcpA (for leptospiral complement regulator-acquiring protein A). LcpA was shown to be an outer membrane protein by using immunoelectron microscopy, cell surface proteolysis, and Triton X-114 fractionation. The gene coding for LcpA is conserved among pathogenic leptospiral strains. This is the first characterization of a Leptospira surface protein that binds to the human complement regulator C4BP in a manner that allows this important regulator to control complement system activation mediated either by the classical pathway or by the lectin pathway. This newly identified protein may play a role in immune evasion by Leptospira spp. and may therefore represent a target for the development of a human vaccine against leptospirosis.

Immune Evasion of Leptospira Species by Acquisition of Human Complement Regulator C4BP

Leptospirosis is a spirochetal zoonotic disease of global distribution with a high incidence in tropical regions. In the last 15 years it has been recognized as an important emerging infectious disease due to the occurrence of large outbreaks in warm-climate countries and, occasionally, in temperate regions. Pathogenic leptospires efficiently colonize target organs after penetrating the host. Their invasiveness is attributed to the ability to multiply in blood, adhere to host cells, and penetrate into tissues. Therefore, they must be able to evade the innate host defense. The main purpose of the present study was to evaluate how several Leptospira strains evade the protective function of the complement system. The serum resistance of six Leptospira strains was analyzed. We demonstrate that the pathogenic strain isolated from infected hamsters avoids serum bactericidal activity more efficiently than the culture-attenuated or the nonpathogenic Leptospira strains. Moreover, both the alternative and the classical pathways of complement seem to be responsible for the killing of leptospires. Serum-resistant and serum-intermediate strains are able to bind C4BP, whereas the serum-sensitive strain Patoc I is not. Surface-bound C4BP promotes factor I-mediated cleavage of C4b. Accordingly, we found that pathogenic strains displayed reduced deposition of the late complement components C5 to C9 upon exposure to serum. We conclude that binding of C4BP contributes to leptospiral serum resistance against host complement.

Exercise does not alter subcellular localization, but increases phosphorylation of insulin-signaling proteins in human skeletal muscle

The subcellular localization of insulin signaling proteins is altered by various stimuli such as insulin, insulin-like growth factor I, and oxidative stress and is thought to be an important mechanism that can influence intracellular signal transduction and cellular function. This study examined the possibility that exercise may also alter the subcellular localization of insulin signaling proteins in human skeletal muscle. Nine untrained males performed 60 min of cycling exercise (~67% peak pulmonary O₂ uptake). Muscle biopsies were sampled at rest, immediately after exercise, and 3 h postexercise. Muscle was fractionated by centrifugation into the following crude fractions: cytosolic, nuclear, and a high-speed pellet containing membrane and cytoskeletal components. Fractions were analyzed for protein content of insulin receptor, insulin receptor substrate (IRS)-1 and -2, p85 subunit of phosphatidylinositol 3-kinase, Akt, and glycogen synthase kinase-3 (GSK-3). There was no significant change in the protein content of the insulin signaling proteins in any of the crude fractions after exercise or 3 h postexercise. Exercise had no significant effect on the phosphorylation of IRS-1 Tyr612 in any of the fractions. In contrast, exercise increased (P < 0.05) the phosphorylation of Akt Ser⁴⁷³ and GSK-3α/ß Ser^9/21 in the cytosolic fraction only. In conclusion, exercise can increase phosphorylation of downstream insulin signaling proteins specifically in the cytosolic fraction but does not result in changes in the subcellular localization of insulin signaling proteins in human skeletal muscle. Change in the subcellular protein localization is therefore an unlikely mechanism to influence signal transduction pathways and cellular function in skeletal muscle after exercise.

Influence of preexercise muscle glycogen content on transcrptional activity of metabolic and myogenic genes in well-trained humans

To determine whether preexercise muscle glycogen content influences the transcription of several early-response genes involved in the regulation of muscle growth, seven male strength-trained subjects performed one-legged cycling exercise to exhaustion to lower muscle glycogen levels (Low) in one leg compared with the leg with normal muscle glycogen (Norm) and then the following day completed a unilateral bout of resistance training (RT). Muscle biopsies from both legs were taken at rest, immediately after RT, and after 3 h of recovery. Resting glycogen content was higher in the control leg (Norm leg) than in the Low leg (435 ± 87 vs. 193 ± 29 mmol/kg dry wt; P < 0.01). RT decreased glycogen content in both legs (P < 0.05), but postexercise values remained significantly higher in the Norm than the Low leg (312 ± 129 vs. 102 ± 34 mmol/kg dry wt; P < 0.01). GLUT4 (3-fold; P < 0.01) and glycogenin mRNA abundance (2.5-fold; not significant) were elevated at rest in the Norm leg, but such differences were abolished after exercise. Preexercise mRNA abundance of atrogenes was also higher in the Norm compared with the Low leg [atrogin: 14-fold, P < 0.01; RING (really interesting novel gene) finger: 3-fold, P < 0.05] but decreased for atrogin in Norm following RT (P < 0.05). There were no differences in the mRNA abundance of myogenic regulatory factors and IGF-I in the Norm compared with the Low leg. Our results demonstrate that 1) low muscle glycogen content has variable effects on the basal transcription of select metabolic and myogenic genes at rest, and 2) any differences in basal transcription are completely abolished after a single bout of heavy resistance training. We conclude that commencing resistance exercise with low muscle glycogen does not enhance the activity of genes implicated in promoting hypertrophy.

Effects of starvation and water quality on the purging process of farmed Murray cod ( Maccullochella peelii peelii )

The aim of this study was to determine the effects of starvation and water quality during the purging process on the biometric parameters, fatty acids, and flavor volatiles of Murray cod farmed in a recirculation system. Market size Murray cod, at the end of the grow-out stage, were divided into eight treatments. The treatments were either fed/starved (F or S) and kept in clean water (CW: CWF2, CWS2, CWF4, and CWS4) or fed/starved and kept in recycled water (RW: RWF2, RWS2, RWF4, and RWS4) for either 2 or 4 weeks. Fish were sampled at 0, 2, and 4 week intervals. Food deprivation was responsible for a significant (P < 0.05) weight loss compared to that of fed treatments. The same was observed for the condition factor (K), hepatosomatic index (HSI), and dress-out percentage (DP). No significant changes were, however, observed in the visceral fat index (VFI). Saturated fatty acids (SFA) were highest in RWF4 and lowest in CWS4 (P < 0.05), while monounsaturated fatty acids (MUFA) were lowest in CWF4 (P < 0.05). Starvation did not affect the flavor volatile compounds, which were mainly affected by changes in water quality. Specifically, total aldehyde (% w/w) content was significantly (P < 0.05) affected by water quality, but the time of purging was not responsible for any noteworthy differences. This study was able to separate the effects of starvation and water quality, in the purging process, on the final eating quality of farmed market size Murray cod. It is concluded that because of the inevitable weight loss during starvation, Murray cod should be fed during the purging stage but kept in clean water and deprived of food only for the time necessary to empty the gastro-intestinal tract.

Thermal degradation of natural rubber/ZNO nanocomposites

Nanoparticies have been widely used to enhance the properties of natural rubber (NR). In the present paper a novel nanocomposite was developed by blending nano-ZnO slurry with prevulcanized NR latex, and the thermal degradation process of pure NR and NR/ZnO nanocomposites with different nano-ZnO loading was studied with a Perkin Elemer TGA-7 thermogravimetric analyzer. The thermal degradation parameters of NR/ZnO (2 parts ZnO per hundred dlY rubber) at different heating rates (Bs) were studied. The results show that the thermal degradation of pure NR and NR/ZnO nanocomposites in nitrogen is a one-step reaction. The degradation temperatures of NR/ZnO nanocomposite increase with an increasing B. The peak height (R_p) on the differential thermogravimetric curve increases with the increase of B. The degradation rates are not affected significantly by B, and the average values of thermal degradation rate C_p and C_f are 44.42 % and 81.04 %, respectively. The thermal degradation kinetic parameters are calculated with Ozawa-Flynn-Wall method. The activation energy (E) and the frequency factor (A) vary with ecomposition degree, and can be divided into three phases corresponding to the volatilization of low-molecular-weight materials, the thermal degradation ofNR main chains and the decomposition of residual carbon.

Physical activity for cancer survivors : a meta-analysis of randomised controlled trials

Objective To systematically evaluate the effects of physical activity in adult patients after completion of main treatment related to cancer. Design Meta-analysis of randomised controlled trials with data extraction and quality assessment performed independently by two researchers. Data sources Pubmed, CINAHL, and Google Scholar from the earliest possible year to September 2011. References from meta-analyses and reviews. Study selection Randomised controlled trials that assessed the effects of physical activity in adults who had completed their main cancer treatment, except hormonal treatment. Results There were 34 randomised controlled trials, of which 22 (65%) focused on patients with breast cancer, and 48 outcomes in our meta-analysis. Twenty two studies assessed aerobic exercise, and four also included resistance or strength training. The median duration of physical activity was 13 weeks (range 3-60 weeks). Most control groups were considered sedentary or were assigned no exercise. Based on studies on patients with breast cancer, physical activity was associated with improvements in insulin-like growth factor-I, bench press, leg press, fatigue, depression, and quality of life. When we combined studies on different types of cancer, we found significant improvements in body mass index (BMI), body weight, peak oxygen consumption, peak power output, distance walked in six minutes, right handgrip strength, and quality of life. Sources of study heterogeneity included age, study quality, study size, and type and duration of physical activity. Publication bias did not alter our conclusions. Conclusions Physical activity has positive effects on physiology, body composition, physical functions, psychological outcomes, and quality of life in patients after treatment for breast cancer. When patients with cancer other than breast cancer were also included, physical activity was associated with reduced BMI and body weight, increased peak oxygen consumption and peak power output, and improved quality of life.

Construção do teste de aptidão criativa - TAC: forma experimental

Este trabalho tem por finalidade a construção de um instrumento psicológico de medida de comportamentos criativos, o Teste de Aptidão Criativa - TAC. A construção do TAC é justificada de um lado, por sua utilidade; sobretudo para o orientador profissional e psicólogo escolar e de outro; pela inexistência no Brasil; em particular no Nordeste, de instrumentos de medida similar. A formulação do problema e seu contexto são descritos logo no início do trabalho, bem como, a fundamentação teórica. Neste particular, enfatizou-se o enfoque psicológico da criatividade v em suas abordagens personológica e cognitiva. Atenção especial foi dada ao estudo do "modelo da estrutura do intelecto" de J. P. Guilford, suporte teórico do TAC. O teste de Aptidão Criativa é apresentado em todas as etapas de sua construção , desde a forma pré-piloto ç à piloto e à experimental, salientando-se seus respectivos subtestes figural e verbal. Ilustrações dos itens, bem como, das formas do teste são também apresentadas para melhor compreensão do texto. Em prosseguimento descreve- se uma pesquisa empírica realizada com o TAC, cujas hipóteses operacionais visam comprovar sua validade de constrito sua fidedignidade. A população foi constituída de alunos, de ambos os sexos, que cursaram em 1977 a 8a. série do 1º grau, de escolas públicas ou particulares; do município do Recife. O trabalho descreve detalhadamente a determinação da amostra, os instrumentos utilizados bem como os critérios de avaliação dos dados. O tratamento estatístico consistiu em medidas de tendência ceutra e de variabilidade; análise fatorial (técnica dos componentes principais-solução Varimax) para verificação da validade de construto. Os resultados obtidos, pela análise fatorial, demonstraram a presença de três fatores: um denominado C e interpretado como Convergente e dois, mensuráveis pelo TAC (conforme mostram as cargas fatoriais) que foram chamados de F (Fluência e Flexibilidade) e 0 Originalidade. O índice de homogeneidade calculado pelo “ de Cronbach" da característica originalidade entendida como essencial para a criatividade foi de 0,72. Tal resultado permite afirmar que na amostra testada o TAC , neste aspecto, apresenta fidedignidade significativa. Uma análise de regressão múltipla por passos foi levada a efeito, visando a identificação dos itens que mais contribuíram para explicação do escore total. Os resultados permitiram apontar uma melhor forma para o TAC figural constituída pelos itens I , II, V, VI , VII e VIII, os quais comporão a forma definitiva do TAC. Calculou-se também o estatístico Z do "Teste de urna Amostra de Kolmogorou para verificação da normalidade da distribuição das três características que o de medir: Fluência, Flexibilidade e Originalidade. Explicita-se ainda uma crítica à técnica de computação das respostas do TAC em face do índice de flexibilidade e propõe-se a realização de estudos, que permitam a formulação de novos procedimentos para um tratamento mais adequado das respostas aos estímulos do teste. Sugere-se também um estudo teórico sistemático de maior profundidade sobre os pontos de convergência entre as teorias psicológicas que tratam o tema para uma melhor compreensão da natureza e dinâmica do comportamento criativa. Propõe-se ainda a continuidade da pesquisa empírica que deu origem a este trabalho, tendo por objetivo imediato a padronização do TAC, de modo a poder ser ele instrumento útil ao psicólogo, na prática da psicologia aplicada.

Avaliação dos impactos de movimentos inesperados nas variáveis macroeconômicas no retorno setorial

O objetivo dessa dissertação é analisar o impacto dos movimentos inesperados de variáveis macroeconômicas nos retornos das ações de empresas de diferentes setores. As variáveis macroeconômicas estudadas serão: produto, juros, inflação e preço de commodities. Estudam-se alguns modelos através de diferentes técnicas de regressão para se chegar àquele que tem a melhor especificação. Com o objetivo de melhorar o poder de explicação dos modelos são utilizados os três fatores de Fama e French como variáveis explicativas e outro modelo que além dos fatores de Fama e French incluiu também momentum. Procura-se analisar a magnitude do impacto dos movimentos inesperados das variáveis macroeconômicas e suas relevâncias estatísticas a cada setor.

Fos-like immunoreactivity in central nervous system of mice simultaneously exposed to the elevated plus-maze and nociception

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

Plasma progesterone concentration in beef heifers receiving exogenous glucose, insulin, or bovine somatotropin

Three experiments were conducted to evaluate plasma concentrations of glucose, insulin, IGF-I, and progesterone (P4) in pubertal beef heifers receiving exogenous glucose, insulin, or sometribove zinc. All heifers used had no luteal P4 synthesis but received a controlled internal drug-releasing device containing 1.38 g of P4 to estimate treatment effects on hepatic P4 degradation. In Exp. 1, 8 pubertal, nulliparous Angus x Hereford heifers (initial BW = 442 +/- 14 kg; initial age = 656 +/- 7 d) were randomly assigned to receive, in a crossover design containing 2 periods of 10 h, intravenous (i.v.) infusions (10 mL) of insulin (1 mu g/kg of BW; INS) or saline (0.9%; SAL). Treatments were administered via jugular venipuncture in 7 applications (0.15 mu g insulin/kg BW per application) 45 min apart (from 0 to 270 min). Blood samples were collected immediately before each infusion as well as at -120, -60, 330, 390, and 450 min relative to the first infusion. Heifers receiving INS had greater (P < 0.01) plasma insulin, reduced (P <= 0.04) plasma glucose and IGF-I, and similar (P = 0.62) plasma P4 concentrations compared with SAL heifers. In Exp. 2, the same heifers were assigned to receive, in a similar experimental design as Exp. 1, i.v. infusions (10 mL) of 1) insulin (1 mu g/kg BW) and glucose (0.5 g/kg BW; INS+G) or 2) SAL. Heifers receiving INS+G had greater (P <= 0.02) plasma insulin, glucose, and P4 but reduced (P = 0.01) plasma IGF-I concentrations compared with SAL heifers. In Exp. 3, the same heifers were assigned to receive, in a crossover design containing 2 periods of 14 d, subcutaneous (s.c.) injections of 1) 250 mg of sometribove zinc (BST) or 2) SAL. Blood samples were collected 3 h apart (0900, 1200, 1500, and 1800 h) from heifers on d 6, 8, and 10 relative to treatment administration (d 1). Heifers receiving BST had greater (P < 0.01) plasma glucose and IGF-I and similar (P >= 0.67) plasma insulin and P4 concentrations compared with SAL heifers. Results from this series of experiments suggested that concurrent increases in glucose and insulin are required to reduce hepatic catabolism and increase plasma concentrations of P4 in bovine females.