Losses of immunoreactive parvalbumin amacrine and immunoreactive alphaprotein kinase C bipolar cells caused by methylmercury chloride intoxication in the retina of the tropical fish Hoplias malabaricus

To quantify the effects of methylmercury (MeHg) on amacrine and on ON-bipolar cells in the retina, experiments were performed in MeHg-exposed groups of adult trahiras (Hoplias malabaricus) at two dose levels (2 and 6 µg/g, ip). The retinas of test and control groups were processed by mouse anti-parvalbumin and rabbit anti-_aprotein kinase C (_aPKC) immunocytochemistry. Morphology and soma location in the inner nuclear layer were used to identify immunoreactive parvalbumin (PV-IR) and _aPKC (_aPKC-IR) in wholemount preparations. Cell density, topography and isodensity maps were estimated using confocal images. PV-IR was detected in amacrine cells in the inner nuclear layer and in displaced amacrine cells from the ganglion cell layer, and _aPKC-IR was detected in ON-bipolar cells. The MeHg-treated group (6 µg/g) showed significant reduction of the ON-bipolar _aPKC-IR cell density (mean density = 1306 ± 393 cells/mm²) compared to control (1886 ± 892 cells/mm²; P < 0.001). The mean densities found for amacrine PV-IR cells in MeHg-treated retinas were 1040 ± 56 cells/mm² (2 µg/g) and 845 ± 82 cells/mm² (6 µg/g), also lower than control (1312 ± 31 cells/mm²; P < 0.05), differently from the data observed in displaced PV-IR amacrine cells. These results show that MeHg changed the PV-IR amacrine cell density in a dose-dependent way, and reduced the density of _aKC-IR bipolar cells at the dose of 6 µg/g. Further studies are needed to identify the physiological impact of these findings on visual function.

Effects of different intensities of physical exercise on insulin sensitivity and protein kinase B/Akt activity in skeletal muscle of obese mice

Objetivo: Investigar os efeitos do exercício físico agudo com diferentes intensidades sobre a sensibilidade à insulina e a atividade da proteína quinase B/Akt no músculo esquelético de camundongos obesos. Método: Foram utilizados camundongos Swiss, divididos aleatoriamente em quatro grupos, que receberam dieta padrão (grupo controle) ou dieta hiperlipídica (grupos obeso sedentário e grupos obesos exercitados 1 e 2), por período de 12 semanas. Dois diferentes protocolos de exercício foram utilizados: natação durante 1 hora com ou sem sobrecarga de 5% da massa corporal. O teste de tolerância à insulina foi realizado para estimar a sensibilidade à insulina. E os níveis protéicos da proteína quinase B/Akt e de sua fosforilação foram determinados no músculo esquelético dos camundongos, através da técnica de Western blot. Resultado: Uma sessão de exercício físico foi capaz de inibir a resistência à insulina em decorrência de uma dieta hiperlipídica. Foi possível demonstrar um aumento na fosforilação da proteína quinase B/Akt, melhora da sinalização da insulina e redução da glicemia de jejum nos camundongos que realizaram 1 hora de natação sem sobrecarga adicional e nos camundongos que realizaram 1 hora de natação com sobrecarga adicional de 5% de sua massa corporal. Entretanto, não houve diferença significativa entre os grupos que realizaram o exercício em diferentes intensidades. Conclusão: Independente da intensidade, o exercício físico aeróbio conseguiu aumentar a sensibilidade à insulina e a fosforilação da proteína quinase B/Akt, revelando ser uma boa forma de tratamento e prevenção do diabetes tipo 2.

Design sistêmico e ecoinovação em APLs: o método F.L.O.R.A como estratégia sustentável aplicada para o polo de confecções de Pernambuco

Pós-graduação em Design - FAAC

Antifungal activity of flavonoids from Heteropterys byrsonimifolia and a commercial source against Aspergillus ochraceus: In silico interactions of these compounds with a protein kinase

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identification of Novel Compounds Inhibiting Chikungunya Virus-Induced Cell Death by High Throughput Screening of a Kinase Inhibitor Library

Chikungunya virus (CHIKV) is a mosquito-borne arthrogenic alphavirus that causes acute febrile illness in humans accompanied by joint pains and in many cases, persistent arthralgia lasting weeks to years. The re-emergence of CHIKV has resulted in numerous outbreaks in the eastern hemisphere, and threatens to expand in the foreseeable future. Unfortunately, no effective treatment is currently available. The present study reports the use of resazurin in a cell-based high-throughput assay, and an image-based high-content assay to identify and characterize inhibitors of CHIKV-infection in vitro. CHIKV is a highly cytopathic virus that rapidly kills infected cells. Thus, cell viability of HuH-7 cells infected with CHIKV in the presence of compounds was determined by measuring metabolic reduction of resazurin to identify inhibitors of CHIKV-associated cell death. A kinase inhibitor library of 4,000 compounds was screened against CHIKV infection of HuH-7 cells using the resazurin reduction assay, and the cell toxicity was also measured in non-infected cells. Seventy-two compounds showing >= 50% inhibition property against CHIKV at 10 mu M were selected as primary hits. Four compounds having a benzofuran core scaffold (CND0335, CND0364, CND0366 and CND0415), one pyrrolopyridine (CND0545) and one thiazol-carboxamide (CND3514) inhibited CHIKV-associated cell death in a dose-dependent manner, with EC50 values between 2.2 mu M and 7.1 mu M. Based on image analysis, these 6 hit compounds did not inhibit CHIKV replication in the host cell. However, CHIKV-infected cells manifested less prominent apoptotic blebs typical of CHIKV cytopathic effect compared with the control infection. Moreover, treatment with these compounds reduced viral titers in the medium of CHIKV-infected cells by up to 100-fold. In conclusion, this cell-based high-throughput screening assay using resazurin, combined with the image-based high content assay approach identified compounds against CHIKV having a novel antiviral activity -inhibition of virus-induced CPE - likely by targeting kinases involved in apoptosis.

Exercise training decreases mitogen-activated protein kinase phosphatase-3 expression and suppresses hepatic gluconeogenesis in obese mice

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

Tonic modulation of anxiety-like behavior by corticotropin-releasing factor (CRF) type 1 receptor (CRF1) within the medial prefrontal cortex (mPFC) in male mice: Role of protein kinase A (PKA)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Role of protein kinase A signaling pathway in cyclosporine nephrotoxicity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Recursos hídricos sob influência de efluentes industriais do polo industrial de Santa Gertrudes

Efluentes industriais e domésticos podem conter agentes químicos e biológicos que, em elevadas concentrações, causam danos aos ecossistemas aquáticos e à saúde ambiental. Um dos efeitos mais nocivos desses agentes poluidores é a sua capacidade de induzir alterações celulares. O uso de testes específicos capazes de detectar o potencial tóxico de substâncias químicas caracteriza-se em uma importante estratégia para avaliação ambiental. Neste contexto, o presente estudo teve como objetivo avaliar os potenciais citotóxico, genotóxico e mutagênico de amostras de água de recursos hídricos da cidade de Santa Gertrudes-SP, relacionada com recebimento de efluentes de atividade ceramista, por meio de ensaios realizados com o sistema-teste de Allium cepa. Foram realizados testes com amostras de água coletadas no córrego Itaqui, após despejos de efluente de indústria cerâmica e em duas nascentes, tanto em período chuvoso como de seca. Para a análise dos parâmetros de toxicidade descritos acima, sementes de A. cepa foram expostas à germinação nas amostras das águas coletadas. Quando as radículas atingiram 2 cm de comprimento, os meristemas foram fixados em solução de Carnoy 3:1, para serem utilizadas nas avaliações das possíveis alterações dos índices de germinação e mitóticos, bem como de indução de aberrações nucleares e cromossômicas. Além disso, também foram avaliadas, como outro parâmetro indicativo de mutagenicidade, as frequências de micronúcleos em células F1 de A. cepa. O controle negativo foi realizado em água (osmose reversa) e o controle positivo em metilmetanosulfonato (MMS). O material fixado foi corado pela reação de Feulgen e as lâminas foram preparadas, utilizando as porções meristemática e F1 das raízes de A. cepa. As análises foram realizadas em microscópio de luz, por meio da contagem de alterações nucleares (micronúcleos e brotos) e de aberrações cromossômicas...

Proposta para melhoria da qualidade dos produtos cerâmicos e de criação de um polo cerâmico artesanal na cidade de Indiana - SP

This work presents a proposal to create a Polo Ceramic Craft in the town of Indiana - SP, through the potter’s organization in a cooperative that will coordinate activities to add value to ceramic pieces. To achieve this, two things are essential: improving the ceramic body and improve the properties of the ceramic material. For the first action it’s necessary to create a Central Mass Production of Ceramics, to provide raw materials and homogeneous composition that results in differentiated ceramic after burning process (sintering). To this end, we propose the incorporation of additives (which act as fluxes) to the clay material. These additives can be mineral such as feldspar and nefelinas or leavings, such as glass powder obtained from disposable containers. For the second action is necessary to acquire an oven, electric or gas, it reaches higher temperatures (around 1200 ° C). The presence of the additive and burning at higher temperatures will enable better production of sintered ceramic material with less porosity and water absorption and higher mechanical strength, and pieces vitrified and glazed, allowing them to assign a higher value. For the production of these materials (thinner walls) requires a smaller volume of clayey raw materials. Besides benefiting the ceramic pieces, the proposed changes reduce the environmental impact caused by burning wood, since it will be replaced by natural gas (or electricity), and even will reduce the disposal of glass containers in the environment by recycling and incorporating this material in the clay. From a social standpoint, the cooperative is crucial to the viability of the proposed project, to coordinate activities and commercial production, which will result in better wages and profits for companies and consequently for the city and its population

A protein kinase screen of Neurospora crassa mutant strains reveals that the SNF1 protein kinase promotes glycogen synthase phosphorylation

Glycogen functions as a carbohydrate reserve in a variety of organisms and its metabolism is highly regulated. The activities of glycogen synthase and glycogen phosphorylase, the rate-limiting enzymes of the synthesis and degradation processes, respectively, are regulated by allosteric modulation and reversible phosphorylation. To identify the protein kinases affecting glycogen metabolism in Neurospora crassa, we performed a screen of 84 serine/threonine kinase knockout strains. We identified multiple kinases that have already been described as controlling glycogen metabolism in different organisms, such as NcSNF1, NcPHO85, NcGSK3, NcPKA, PSK2 homologue and NcATG1. In addition, many hypothetical kinases have been implicated in the control of glycogen metabolism. Two kinases, NcIME-2 and NcNIMA, already functionally characterized but with no functions related to glycogen metabolism regulation, were also identified. Among the kinases identified, it is important to mention the role of NcSNF1. We showed in the present study that this kinase was implicated in glycogen synthase phosphorylation, as demonstrated by the higher levels of glycogen accumulated during growth, along with a higher glycogen synthase (GSN) ±glucose 6-phosphate activity ratio and a lesser set of phosphorylated GSN isoforms in strain Ncsnf1KO, when compared with the wild-type strain. The results led us to conclude that, in N. crassa, this kinase promotes phosphorylation of glycogen synthase either directly or indirectly, which is the opposite of what is described for Saccharomyces cerevisiae. The kinases also play a role in gene expression regulation, in that gdn, the gene encoding the debranching enzyme, was down-regulated by the proteins identified in the screen. Some kinases affected growth and development, suggesting a connection linking glycogen metabolism with cell growth and development.

A influência da temperatura na secagem forçada da matéria-prima cerâmica da Formação Corumbataí na região do Polo de Santa Gertrudes (SP)

Pós-graduação em Geociências e Meio Ambiente - IGCE

Silencing mitogen-activated protein kinase-activated protein kinase-2 arrests inflammatory bone loss

p38 mitogen-activated protein kinases (MAPKs) are critical for innate immune signaling and subsequent cytokine expression in periodontal inflammation and bone destruction. In fact, previous studies show that systemic p38 MAPK inhibitors block periodontal disease progression. However, development of p38 MAPK inhibitors with favorable toxicological profiles is difficult. Here, we report our findings regarding the contribution of the downstream p38 MAPK substrate, mitogen-activated protein kinase-activated protein kinase 2 (MK2 or MAPKAPK-2), in immune response modulation in an experimental model of pathogen-derived lipopolysaccharide (LPS)-induced periodontal bone loss. To determine whether small interfering RNA (siRNA) technology has intraoral applications, we initially validated MK2 siRNA specificity. Then, gingival tissue surrounding maxillary molars of rats was injected with MK2 siRNA or scrambled siRNA at the palatal regions of bone loss. Intraoral tissues treated with MK2 siRNA had significantly less MK2 mRNA expression compared with scrambled siRNA-treated tissues. MK2 siRNA delivery arrested LPS-induced inflammatory bone loss, decreased inflammatory infiltrate, and decreased osteoclastogenesis. This proof-of-concept study suggests a novel target using an intraoral RNA interference strategy to control periodontal inflammation.

Avaliação do perfil antropométrico e de força de jovens jogadores de polo-aquático

The present study aimed to analyze the performance of 'boost' among water polo athletes. Twelve athletes (13,9  1,08 years, 169,0  6,0 cm and 59,5  13,5 kg of bodyweight) were under went to anthropometric assessment, body composition and maximum concentric strength of lower limbs. The jump test in the water was done by 'boost' and scaled to jump height. According to the results of strength (Leg Press 45: 157,08  43,30 kg Leg Extension: 72,08  12,86 kg), anthropometry (arm circumf.: 26,04  3,25 cm, thigh circumf.: 50,47  7,65 cm, sholder circumf.: 95,97  8,00 cm, elbow diam.: 4,74  0,51 cm, wrist diam.: 3,21  0,34 cm, knee diam.: 7,89  0,58 cm, biacromial diam.: 37,49  3,81 cm and bi-iliac diam.: 25,43  4,34 cm) and body composition (BMI: 20,74  4,00 kg / m² and% fat: 8,45  4,28), only stature showed a linear correlation (r= 0,595) to the values of vertical jump (height: 47,80  4,35 cm) performance. This anthropometric variable was one able to influence the height of performance by modifying it self, perhaps by maturity. However maturity may be an implicity feature of performance, since force is a concerned effect of the growth. Otherwise technique could play a mains role to the performance of the 'boost'. It can be conclude that possibly is advantageous for water polo athlete be higher and more mature for the realization of 'boost', and the training directed for the technique, but is necessary muscle fitness for the holder