Immunolocalization of nitric oxide synthase isoforms in human archival and rat tissues, and cultured cells

Nitric oxide (NO) exerts important physiological and pathological roles in humans. The study of NO requires the immunolocalization of its synthesizing enzymes, neuronal, endothelial and inducible NO synthases (NOS). NOS are labile to formalin-fixation and paraffin-embedding, which are used to prepare human archival tissues. This lability has made NOS immunohistochemical studies difficult, and a detailed protocol is not yet available. We describe here a protocol for the immunolocalization of NOS isoforms in human archival cerebellum and non-nervous tissues, and in rat tissues and cultured cells. Neuronal NOS antigenicity in human archival and rat nervous tissue sections was microwave-retrieved in 50 mM Tris-HCl buffer, pH 9.5, for 20 min at 900W. Neuronal NOS was expressed in stellate, basket, Purkinje and granule cells in human and rat cerebellum. Archival and frozen human cerebellar sections showed the same neuronal NOS staining pattern. Archival cerebellar sections not subjected to antigen retrieval stained weakly. Antigenicity of inducible NOS in human lung was best retrieved in 10 mM sodium citrate buffer, pH 6.0, for 15 min at 900W. Inflammatory cells in a human lung tuberculoma were strongly stained by anti-inducible NOS antibody. Anti-endothelial NOS strongly stained kidney glomeruli. Cultured PC12 cells were strongly stained by anti-neuronal NOS without antigen retrieving. The present immunohistochemistry protocol is easy to perform, timeless, and suitable for the localization of NOS isoforms in nervous and non-nervous tissues, in human archival and rat tissues. It has been extensively used in our laboratory, and is also appropriate for other antigens. (C) 2011 Elsevier B.V. All rights reserved.

Serological survey of Toxoplasma gondu in captive Neotropical felids from Southern Brazil

Toxoplasma gondu is the causative intracellular protozoan of toxoplasmosis inhuman being and animals Members of the Felidae family are considered the single definitive host for the infection both wild and domestic cats are able to excrete oocysts in the environment Wild cats maintained in captivity may serve as source of infection for other clinically susceptible animals in the same environment The aim of this study was to determine the frequency of T gondu IgG antibodies in 57 neotropical felids (1 Leopardus geoffroyi 3 Puma yagouaroundi 17 Leopard us wiedu 22 Leopardus tigrinus and 14 Leopard us pardalis) kept at the Bela Vista Biological Sanctuary Itaipu Binacional Southern Brazil by the modified agglutination test (MAT) using titer 16 as cut-off point Seropositivity was observed in 38/57 (66 67% 95% CI 53 66-77 51%) samples with higher frequency in ocelots (71 43%) Wild-caught felids were three times more likely to be infected when compared to zoo-born animals (P <= 0 05) and age of wild-caught animals (P= 0 6892 95% CI = 0 7528-166) was not significant as a risk factor for the infection the same occurring with zoo-born animals (P=0 05 95% CI = 06267-24052) These results suggest that despite efforts to control T gondu infection in zoo facilities such as individual pens hygiene monitoring veterinary care and pre-frozen meat offered as food non-domestic feuds kept in captivity particularly the wild-caught specimens may be invariably exposed to infection due to other environmental sources (C) 2010 Elsevier B V All rights reserved

Influence of etching time on bond strength in dentin irradiated with erbium lasers

The purpose of this in vitro study was to evaluate the effect of etching time on the tensile bond strength (TBS) of a conventional adhesive bonded to dentin previously irradiated with erbium:yttrium-aluminum-garnet (Er:YAG) and erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) lasers. Buccal and lingual surfaces of 45 third molars were flattened until the dentin was exposed and randomly assigned to three groups (n = 30) according to the dentin treatment: control (not irradiated), irradiated with Er:YAG (1 W; 250 mJ; 4 Hz; 80.6 J/cm(2)) laser or Er,Cr:YSGG (4 W; 200 mJ; 20 Hz; 71.4 J/cm(2)) laser, and into three subgroups (n = 10) according to acid etching time (15 s, 30 s or 60 s) for each experimental group. After acid etching, the adhesive was applied, followed by the construction of an inverted cone of composite resin. The samples were immersed in distilled water (37A degrees C for 24 h) and subjected to TBS test [50 kilogram-force (kgf), 0.5 mm/min]. Data were analyzed by analysis of variance (ANOVA) and Tukey statistical tests (P a parts per thousand currency signaEuro parts per thousand 0.05). Control group samples presented significant higher TBS values than those of all lased groups. Both irradiated groups exhibited similar TBS values. Samples subjected to the different etching times in each experimental group presented similar TBS. Based on the conditions of this in vitro study we concluded that Er:YAG and Er,Cr:YSGG laser irradiation of the dentin weakens the bond strength of the adhesive. Moreover, increased etching time is not able to modify the bonding strength of the adhesive to irradiated dentin.

Effects of Nd:YAG Laser Irradiation on the Hybrid Layer of Different Adhesive Systems

Purpose: The aim of this in vitro study was to evaluate the microtensile bond strength (mu TBS) and hybrid layer morphology of different adhesive systems, either followed by treatment with Nd:YAG laser irradiation or not. Previous studies have shown the effects of Nd:YAG laser irradiation on the dentin surface at restoration margins, but there are few reports about the significance of the irradiation on the hybrid layer. Materials and Methods: The flattened coronal and root dentin samples of 24 bovine teeth were randomly divided into 8 groups, according to the adhesive system used - Scotchbond Multi Purpose (SBMP) or Clearfil SE Bond (CSEB) - and were either irradiated with Nd:YAG or not, with different parameters: 0.8 W/10 Hz, 0.8 W/20 Hz, 1.2 W/10 Hz, 1.2 W/20 Hz. The left sides of specimens were the control groups, and right sides were irradiated. A composite crown was built over bonded surfaces and stored in water (24 h at 37 degrees C). Specimens were sectioned vertically into slabs that were subjected to mu TBS testing and observed by SEM. Results: Control groups (27.81 +/- 1.38) showed statistically higher values than lased groups (21.37 +/- 0.99), and CSEB control group values (31.26 +/- 15.71) were statistically higher than those of SBMP (24.3 +/- 10.66). There were no significant differences between CSEB (20.34 +/- 10.01) and SBMP (22.43 +/- 9.82) lased groups. Among parameters tested, 0.8 W/10 Hz showed the highest value (25.54 +/- 11.74). Nd:YAG laser irradiation caused dentin to melt under the adhesive layer of both adhesive systems tested. Conclusion: With the parameters used in this study, Nd:YAG laser irradiation of the hybrid layer promoted morphological changes in dentin and negatively influenced the bond strength of both adhesive systems.

Effect of defocused infrared diode laser on salivary flow rate and some salivary parameters of rats

This study aims to investigate whether infrared diode low-level laser therapy (LLLT) increased salivary flow rate and altered pH value, protein concentration, and peroxidase and amylase activities in saliva of rats. Wistar rats were used and divided into three groups. Experimental groups (A and 13) had their parotid, submandibular and sublingual glands submitted to diode laser, 808-nm wavelength, on two consecutive days. The dose results were 4 and 8 J/cm(2), respectively. A red guide light was used to visualize the irradiated area. Group C was irradiated only with red pilot beam and served as control. The saliva samples were collected after each irradiation step (first and second collection days) and 1 week after the first irradiation (seventh day). Statistical analysis was performed, and differences were observed according to different days of salivary collection. The results showed that salivary flow rate for groups A and B was higher on the seventh day if it is compared to data obtained for the first day (p<0.05). LLLT applications on salivary glands are a therapy procedure that requires further studies.

Different air-water spray regulations affect the healing of Er,Cr:YSGG laser incisions

Surgeries performed with high-intensity laser devices may be improved with accurate protocols, including the air-water spray regulation. Thus, this study sought to investigate the healing process of wounds made on the dorsum of rat tongues using an Er,Cr:YSGG laser device with different air-water spray regulations. The incisions were made on the dorsum of Wistar rat tongues using an Er,Cr:YSGG laser with three different air-water spray regulations (100/0%, 50/50%, 11/7%). Scalpel incisions functioned as controls. The sacrifices occurred between 0 and 14 days after surgery. Morphological, histological, and immunohistochemical (fibronectin and type III collagen) analysis of the wounds were performed. The air-water spray regulation influenced wound healing and the inflammatory response, especially in the earlier stages. Incisions performed using the 100/0% air/water spray regulation had the worst results, expressing a greater amount of fibronectin and type III collagen. The 50/50% air/water spray regulation brought in a non-clear surgical field and poor laser interaction with the tissue. The 11/7% air/water spray regulation showed the best clinical results and less pronounced histological events. According to the results encountered, the air-water spray should be regulated to improve surgery.

Influence of Antiplatelet Drugs in the Pathogenesis of Experimental Periodontitis and Periodontal Repair in Rats

Background: Platelets contain an array of biologic mediators that can modulate inflammation and repair processes including proinflammatory mediators and growth factors. Previous studies have shown that periodontitis and periodontal repair are associated with platelet activation. We hypothesized that drug-induced platelet inactivation may interfere in the processes of inflammation and repair in experimental periodontitis in rats by suppressing the release of biologic mediators from platelets to the site of injury. Methods: To measure the effects on periodontitis, ligatures were placed around first molars, and aspirin (Asp, 30 mg/kg) or clopidogrel (Clo, 75 mg/kg) was given intragastrically once daily for 15 days. Interleukin-6 (IL-6), tumor necrosis factor-a (TNF-alpha), and thromboxane A(2) levels were measured by enzyme-linked immunosorbent assay. To evaluate the effects of antiplatelet drugs on periodontal repair, ligatures were removed after 15 days of periodontitis induction, and Asp or Clo were administered beginning the following day for 15 days. Periodontal repair was assessed by microcomputed tomography. Results: On periodontitis phase, Asp and Clo significantly reduced levels of TNF-alpha and II-6 (P < 0.05), but only Asp decreased thromboxane A(2) (P < 0.05). Asp and Clo decreased inflammatory infiltration; however, this reduction was more pronounced with Clo treatment (P < 0.05). Histometric analysis showed that Asp and Clo impaired alveolar bone resorption. During the repair phase and after removal of the ligatures, microcomputed tomography analysis demonstrated that treatment with Asp and Clo did not impair alveolar bone repair. Conclusion: Systemic administration of Asp and Clo attenuates the inflammation associated with periodontitis without affecting the repair process when stimulus is removed. J Periodontol 2011;82:767-777.

In vivo and in vitro models demonstrate a role for caveolin-1 in the pathogenesis of ischaemic acute renal failure

Caveolae and their proteins, the caveolins, transport macromolecules; compartmentalize signalling molecules; and are involved in various repair processes. There is little information regarding their role in the pathogenesis of significant renal syndromes such as acute renal failure (ARF). In this study, an in vivo rat model of 30 min bilateral renal ischaemia followed by reperfusion times from 4 h to 1 week was used to map the temporal and spatial association between caveolin-1 and tubular epithelial damage (desquamation, apoptosis, necrosis). An in vitro model of ischaemic ARF was also studied, where cultured renal tubular epithelial cells or arterial endothelial cells were subjected to injury initiators modelled on ischaemia-reperfusion (hypoxia, serum deprivation, free radical damage or hypoxia-hyperoxia). Expression of caveolin proteins was investigated using immunohistochemistry, immunoelectron microscopy, and immunoblots of whole cell, membrane or cytosol protein extracts. In vivo, healthy kidney had abundant caveolin-1 in vascular endothelial cells and also some expression in membrane surfaces of distal tubular epithelium. In the kidneys of ARF animals, punctate cytoplasmic localization of caveolin-1 was identified, with high intensity expression in injured proximal tubules that were losing basement membrane adhesion or were apoptotic, 24 h to 4 days after ischaemia-reperfusion. Western immunoblots indicated a marked increase in caveolin-1 expression in the cortex where some proximal tubular injury was located. In vitro, the main treatment-induced change in both cell types was translocation of caveolin-1 from the original plasma membrane site into membrane-associated sites in the cytoplasm. Overall, expression levels did not alter for whole cell extracts and the protein remained membrane-bound, as indicated by cell fractionation analyses. Caveolin-1 was also found to localize intensely within apoptotic cells. The results are indicative of a role for caveolin-1 in ARF-induced renal injury. Whether it functions for cell repair or death remains to be elucidated. Copyright (C) 2003 John Wiley Sons, Ltd.

Curso de monitoramento tem??tico do PPA 2012-2015 e acompanhamento or??ament??rio 2013

O curso teve como ementa: apresentar conceitos b??sicos de Monitoramento e Avalia????o; a evolu????o hist??rica da metodologia de monitoramento e avalia????o dos PPAs: 2000-2003; 2004-2007; e 2008-2011; o Monitoramento das Agendas Estrat??gicas de Governo; o monitoramento e avalia????o do PPA 2012-2015: metodologia, concep????o de monitoramento e avalia????o, e suporte informatizado (SIOP); o monitoramento das prioridades da Presid??ncia, dos programas tem??ticos, incluindo os indicadores, os objetivos e as metas do plano; a proposta de monitoramento participativo do PPA 2012-2015, por meio da rela????o do governo com o F??rum Interconselhos de pol??ticas p??blicas; e o acompanhamento das a????es e os fundamentos dos planos or??ament??rios (PO) do Or??amento de 2013

O uso da ren??ncia fiscal na ??rea de museus

O objetivo desse artigo ?? investigar como a ren??ncia fiscal ?? utilizada na ??rea de museus. A fundamenta????o te??rica faz breve apresenta????o da literatura sobre implementa????o e descreve o uso da ren??ncia fiscal na ??rea de museus mediante an??lise de seis vari??veis: incentivos, autonomia, informa????o, apoio institucional, apoio social e etapas e tempo. A metodologia utilizada foi a an??lise documental e entrevistas. A an??lise foi feita com ajuda do programa NVIVO9. As principais conclus??es s??o que embora haja incentivos e autonomia em graus suficientes para permitir boa implementa????o via ren??ncia fiscal, h?? quest??es informacionais e referentes ??s etapas e tempo que dificultam sua implementa????o. Al??m disso, a ren??ncia fiscal ?? concentradora, desigual e apresenta baixa participa????o de empresas do setor privado. Esses problemas parecem diminuir a capacidade de a implementa????o de museus mediante o uso da ren??ncia fiscal alcan??ar os efeitos almejados.

O diagnóstico da linguagem e a linguagem do diagnóstico : uma perspectiva pragmática

A teoria dos atos de fala, de John Austin, apresenta a linguagem a partir de uma perspectiva pragmática, ou seja, quando os signos têm um compromisso com a produção de realidades, de mundo e de sujeitos. A linguagem possui – nessa perspectiva – força de criação. Em sintonia com os estudos da pragmática da linguagem, defenderemos a ideia que uma classificação não é mera descrição neutra acerca dos fatos, e Ian Hacking será a ponte entre a linguagem (enquanto fonte de produção de mundo) e as classificações (como produtoras de sujeitos). Para esse autor, as classificações tanto transformam quanto são transformadas. Elas, de diversas formas, interagem entre si, com os sujeitos, com as instituições, com os saberes, enfim, com tudo aquilo a que faz referência. A partir de uma revisão, análise e composição bibliográfica, o trabalho se dedicará a produzir um elo entre a problemática da linguagem-ato, tal como postulada por John Austin, e o caráter produtor de realidade das classificações. Dentre as classificações, daremos importância particular ao diagnóstico (enquanto uma espécie de estudo de caso na teoria dos enunciados) e seus efeitos pragmáticos, a partir dos elementos que lhe dão força de eficácia e existência em nosso mundo contemporâneo, tendo em vista o aumento do número e da proliferação de categorias nosológicas no corpo social. Trataremos, em particular, as classificações da medicina psiquiátrica. Abordaremos, por um lado, as condições que contribuem na emergência e na potência de um diagnóstico e, por outro, seus efeitos. Dentre os efeitos que daremos importância, encontramos a formação de novos sujeitos e modos de vida. Sendo assim, tanto as condições de eficácia de um enunciado quanto seus efeitos são estendidos à noção de diagnóstico enquanto uma classificação numa condição sociohistórica particular.

Somos quase felizes : movimentos do desejo em tempos de ansiolítico

O presente trabalho versa sobre modos de vida no contemporâneo referente às práticas em torno de medicamentos ansiolíticos. Propomos um modo de pesquisa que não visa a descoberta de verdades, mas a escuta e a escrita das sensibilidades em processo na atualidade. Apropriamo-nos do cinema e da entrevista, intercessores deste trabalho, para dar vazão às vozes que se proferem em torno dessa temática. Os filmes: A pele que habito, Medianeras e O palhaço, somado a duas entrevistas com usuários de ansiolíticos, compõe o corpo desta pesquisa. As análises e a escuta deste campo nos levaram a uma discussão paradoxal entre modos de vida os quais, ao mesmo tempo em que nos lança em ritmos cada vez mais velozes e adoecedores - resultado das forças capitalísticas, tecnológica e midiática – também produzem desejo de anestesiamento do corpo, para suportar a saturação a qual nos encontramos. Corpos que não têm agüentado mais os novos ritmos de vida e, por isso, pedem uma nova ética, uma política de recusa aos ritmos estafantes. Uma discussão com base nos autores Deleuze, Guattari, Nietzsche, Spinoza e Foucault.

Constituição do campo sensível : apegos, renúncias e liberdade – uma contribuição da filosofia budista

Esta dissertação foi construída em interface com a tradição oral milenar da filosofia budista, respeitando um estilo tradicional dos ensinamentos em sua transmissão oral. Por meio da perspectiva de uma "filosofia prática", constitui-se uma metodologia onde as questões se desdobram em reflexões e ideias encadeadas, o que permite contemplar um tema de vários ângulos, tendo por fio norteador os ensinamentos budistas da linhagem Sakya, que orientam uma postura ética para a vida cotidiana. Abordam-se as "orientações para o viver" contidas nos ensinamentos com ênfase no domínio da mente, da motivação e da intenção da ação humana. Para a realização desse exercício ético, que transforma o campo da educação – processo de ensino e de aprendizagem – e o da produção de conhecimento – constituição de saberes sobre si e sobre o mundo –, a prática da meditação se coloca como delineadora de novos contornos para o pensar, o sentir e o agir, o que se evidencia por meio do ensinamento clássico “ Libertando-se dos Quatro Apegos”, referência central para este trabalho. Nesta perspectiva, a meditação é abordada como campo da experiência, como um método com o qual podemos viver a “não separatividade de sujeito-corpo-mente”. A importância da meditação para um processo de criação de experiências constitutivas de um estar desperto. Este despertar se faz por meio de práticas onde “nós somos a experiência da mente”, “nós somos a mente em si”, e a prática da meditação nos propicia essa experiência. A experiência de "estar presente em cada momento" pode nos levar ao domínio de nós mesmos, ao reconhecimento do outro como ser humano que também somos, e ao desenvolvimento da compaixão por todos os seres. Por meio desse processo podemos reconhecer o estado Búdico, ou melhor, o estado de Budeidade que todos temos em potencial. Budha é uma potência em nós, potência de desenvolvermos compaixão equânime para com todos os seres E podemos fazer isso reconhecendo dois aspectos importantes: os quatro apegos que nos aprisionam e a maneira de nos libertarmos deles para trilhar o caminho desperto.