Contribution of peripheral vanilloid receptor to the nociception induced by injection of spermine in mice

Polyamines (putrescine, spermidine and spermine) are important endogenous regulators of ion channels, such as vanilloid (TRPV1), glutamatergic (NMDA or AMPA/kainate) and acid-sensitive (ASIC) receptors. In the present study, we have investigated the possible nociceptive effect induced by polyamines and the mechanisms involved in this nociception in vivo. The subcutaneous (s.c.) injection of capsaicin (as positive control), spermine, spermidine or putrescine produced nociception with ED(50) of 0.16 (0.07-0.39) nmol/paw, 0.4 (0.2-0.7) mu mol/paw, 0.3 (0.1-0.9) mu mol/paw and 3.2 (0.9-11.5) mu mol/paw, respectively. The antagonists of NMDA (MK801, 1 nmol/paw), AMPA/kainate (DNQX, 1 nmol/paw) or ASIC receptors (amiloride, 100 nmol/paw) failed to reduce the spermine-trigged nociception. However, the TRPV1 antagonists capsazepine or SB366791 (1 nmol/paw) reduced spermine-induced nociception, with inhibition of 81 +/- 10 and 68 +/- 9%, respectively. The previous desensitization with resiniferatoxin (RTX) largely reduced the spermine-induced nociception and TRPV1 expression in the sciatic nerve, with reductions of 82 +/- 9% and 67 +/- 11%, respectively. Furthermore, the combination of spermine (100 nmol/paw) and RTX (0.005 fmol/paw), in doses which alone were not capable of inducing nociception, produced nociceptive behaviors. Moreover, different concentrations of spermine (3-300 mu M) enhanced the specific binding of [(3)H](center dot)-RTX to TRPV1 receptor. Altogether, polyamines produce spontaneous nociceptive effect through the stimulation of TRPV1, but not of ionotropic glutamate or ASIC receptors. (C) 2011 Elsevier Inc. All rights reserved.

Molecular cloning and biochemical characterization of a myotoxin inhibitor from Bothrops alternatus snake plasma

Phospholipases A(2) (PLA(2)s) are important components of Bothrops snake venoms, that can induce several effects on envenomations such as myotoxicity, inhibition or induction of platelet aggregation and edema. It is known that venomous and non-venomous snakes present PLA(2) inhibitory proteins (PLIs) in their blood plasma. An inhibitory protein that neutralizes the enzymatic and toxic activities of several PLA2s from Bothrops venoms was isolated from Bothrops alternatus snake plasma by affinity chromatography using the immobilized myotoxin BthTX-I on CNBr-activated Sepharose. Biochemical characterization of this inhibitory protein, denominated alpha BaltMIP, showed it to be a glycoprotein with Mr of similar to 24,000 for the monomeric subunit. CD spectra of the PLA(2)/inhibitor complexes are considerably different from those corresponding to the individual proteins and data deconvolution suggests that the complexes had a relative gain of helical structure elements in comparison to the individual protomers, which may indicate a more compact structure upon complexation. Theoretical and experimental structural studies performed in order to obtain insights into the structural features of aBaltMIP indicated that this molecule may potentially trimerize in solution, thus strengthening the hypothesis previously raised by other authors about snake PLIs oligomerization. (C) 2010 Elsevier Masson SAS. All rights reserved.

The association of Na,K-ATPase subunits studied by circular dichroism, surface tension and dilatational elasticity

Different stoichiometries are observed between alpha and beta subunits of Na,K-ATPase that depend on the method employed to solubilize and purify the enzyme. It is not known whether this variability is due to loss of protein-protein association, or is a result of the replacement of essential phospholipids by detergent molecules. With the aim of understanding the effect of enzyme/surfactant ratio on both the catalytic activity and the enzyme structure, we have investigated the bulk and surface properties of the enzyme. The circular dichroism (CD) spectra, surface tension and dilatational surface elasticity results were compared with the residual ATPase activity of the Na,K-ATPase in different surfactant and protein concentrations. Na,K-ATPase in the (alpha beta)(2) form dissociated to the alpha beta form on dilution, and associated to the (alpha beta)(4) form when concentrated. These different stoichiometries have similar ATPase activities and are in equilibrium at C(12)E(8) concentrations below the CIVIC (0.053 mg mL(-1)). At detergent concentrations above the CIVIC the ATPase activity of all forms was abolished, which is concomitant with the dissociation of the a and subunits. (C) 2008 Elsevier Inc. All rights reserved.

Unraveling the Na,K-ATPase alpha(4) Subunit Assembling Induced by Large Amounts of C(12)E(8) by Means of Small-Angle X-ray Scattering

In the current work, we studied the effect of the nonionic detergent dodecyloctaethyleneglycol, C(12)E(8), on the structure and oligomeric form of the Na,K-ATPase membrane enzyme (sodium-potassium pump) in aqueous suspension, by means of small-angle X-ray scattering (SAXS). Samples composed of 2 mg/mL of Na,K-ATPase, extracted from rabbit kidney medulla, in the presence of a small amount of C(12)E(8) (0.005 mg/mL) and in larger concentrations ranging from 2.7 to 27 mg/mL did not present catalytic activity. Under this condition, an oligomerization of the alpha subunits is expected. SAXS data were analyzed by means of a global fitting procedure supposing that the scattering is due to two independent contributions: one coming from the enzyme and the other one from C(12)E(8) micelles. In the small detergent content (0.005 mg/mL), the SAXS results evidenced that Na,K-ATPase is associated into aggregates larger than (alpha beta)(2) form. When 2.7 mg/mL of C(12)E(8) is added, the data analysis revealed the presence of alpha(4) aggregates in the solution and some free micelles. Increasing the detergent amount up to 27 mg/mL does not disturb the alpha(4) aggregate: just more micelles of the same size and shape are proportionally formed in solution. We believe that our results shed light on a better understanding of how nonionic detergents induce subunit dissociation and reassembling to minimize the exposure of hydrophobic residues to the aqueous solvent.

Association of polymorphisms of glutamate-cystein ligase and microsomal triglyceride transfer protein genes in non-alcoholic fatty liver disease

Background and Aims: Although the metabolic risk factors for non-alcoholic fatty liver disease (NAFLD) progression have been recognized, the role of genetic susceptibility remains a field to be explored. The aim of this study was to examine the frequency of two polymorphisms in Brazilian patients with biopsy-proven simple steatosis or non-alcoholic steatohepatitis (NASH): -493 G/T in the MTP gene, which codes the protein responsible for transferring triglycerides to nascent apolipoprotein B, and -129 C/T in the GCLC gene, which codes the catalytic subunit of glutamate-cystein ligase in the formation of glutathione. Methods: One hundred and thirty-one biopsy-proven NAFLD patients (n = 45, simple steatosis; n = 86, NASH) and 141 unrelated healthy volunteers were evaluated. Genomic DNA was extracted from peripheral blood cells, and the -129 C/T polymorphism of the GCLC gene was determined by restriction fragment length polymorphism (RFLP). The -493 G/T polymorphism of the MTP gene was determined by direct sequencing of the polymerase chain reaction products. Results: The presence of at least one T allele in the -129 C/T polymorphism of the GCLC gene was independently associated with NASH (odds ratio 12.14, 95% confidence interval 2.01-73.35; P = 0.007), whereas, the presence of at least one G allele in the -493 G/T polymorphism of the MTP gene differed slightly between biopsy-proven NASH and simple steatosis. Conclusion: This difference clearly warrants further investigation in larger samples. These two polymorphisms could represent an additional factor for consideration in evaluating the risk of NAFLD progression. Further studies involving a larger population are necessary to confirm this notion.

Genes Differentially Expressed in Prolactinomas Responsive and Resistant to Dopamine Agonists

Background/Aims: Prolactin (PRL) secretion and its gene expression are inhibited by dopamine. Prolactinomas are the most common secreting pituitary adenomas, and dopamine agonists (DA) are the first choice for their treatment. However, a subset of patients is resistant to DA. As the mechanisms involved in DA resistance are not fully understood, the aim of this study was to obtain new insights regarding the molecular differences between the prolactinomas that are responsive to DA and those that are resistant. Methods: Tumor tissue samples were collected from 17 patients who harbored prolactinomas, which were classified as responsive or resistant according to their clinical and laboratorial reaction to DA. The expression of 6 genes was evaluated by real-time polymerase chain reaction: dopamine receptor type 2 (DRD 2), nerve growth factor-beta (NGFB) and its receptor (NGFR), estrogen receptor-alpha (ERA), estrogen receptor-beta (ERB) and the pituitary tumor transforming gene (PTTG). Results: Median DRD 2 and NGFR expression in responsive patients was significantly higher than in resistant ones (p = 0.029 and p = 0.020, respectively). Moreover, the expressions of DRD 2 and NGFR were positively correlated with PRL decrease during treatment (r = 0.66, p = 0.005 and r = 0.57, p = 0.044, respectively). Furthermore, ERB expression was positively correlated to PTTG expression (r = 0.68, p = 0.032) and negatively correlated to NGFB expression (r = -0.75, p = 0.02). Conclusions: DRD2 and NGFR expressions are related to the responsiveness of prolactinoma to DA. However, PTTG, ERB and ERA expressions are not. Also ERB, ERA and PTTG expressions did not present a clear correlation to tumor aggressiveness. Furthermore, the response of prolactinomas to DA should be viewed as a spectrum ranging from the most responsive to the most resistant ones. Copyright (c) 2008 S. Karger AG, Basel

Expression of neurotensin and its receptors in pituitary adenomas

The neurotensin (NT) produced in the hypothalamus and in pituitary gonadotrophs and thyrotrophs participates in neuroendocrine regulation. Recently, the involvement of this peptide in normal and neoplastic cell proliferation has been postulated. In the present study, we evaluated the expression of NT and its receptors (NTR1, 2 and 3) in a series of 50 pituitary adenomas [11 growth hormone (GH)-, eight prolactin (PRL)-, four adrenocorticotrophic hormone (ACTH)- and 27 nonfunctioning adenomas]. NT mRNA expression was significantly higher in functioning compared to nonfunctioning adenomas and with normal pituitary. Nonfunctioning pituitary adenomas showed lower expression of NT mRNA than normal pituitary. In the immunohistochemical study of functioning adenomas, NT was colocalised with GH, PRL and ACTH secreting cells. In nonfunctioning adenomas, the NT immunoreactivity intensity was variable among the samples. NTR3 mRNA expression was observed in all examined samples and was higher in the adenomas, both functioning and nonfunctioning, compared to normal pituitary. By contrast, NTR1 and NTR2 mRNA were not detected in either pituitary adenomas or normal tissue. The higher expression of NTR3, as well as the expression of NT by tumoural corticotrophs, lactotrophs and somatotrophs, which are cells types that do not express this peptide in the normal pituitary, suggests that NT autocrine and/or paracrine stimulation mediated by NTR3 may be a mechanism associated with the tumourigenesis of functioning adenomas.

Endoscopic Anatomy of the Pterygopalatine Fossa and the Transpterygoid Approach: Development of a Surgical Instruction Model

Introduction: The pterygopalatine fossa (PPF) is a narrow space located between the posterior wall of the antrum and the pterygoid plates. Surgical access to the PPF is difficult because of its protected position and its complex neurovascular anatomy. Endonasal approaches using rod lens endoscopes, however, provide better visualization of this area and are associated with less morbidity than external approaches. Our aim was to develop a simple anatomical model using cadaveric specimens injected with intravascular colored silicone to demonstrate the endoscopic anatomy of the PPF. This model could be used for surgical instruction of the transpterygoid approach. Methods: We dissected six PPF in three cadaveric specimens prepared with intravascular injection of colored material using two different injection techniques. An endoscopic endonasal approach, including a wide nasoantral window and removal of the posterior antrum wall, provided access to the PPF. Results: We produced our best anatomical model injecting colored silicone via the common carotid artery. We found that, using an endoscopic approach, a retrograde dissection of the sphenopalatine artery helped to identify the internal maxillary artery (IMA) and its branches. Neural structures were identified deeper to the vascular elements. Notable anatomical landmarks for the endoscopic surgeon are the vidian nerve and its canal that leads to the petrous portion of the internal carotid artery (ICA), and the foramen rotundum, and V2 that leads to Meckel`s cave in the middle cranial fossa. These two nerves, vidian and V2, are separated by a pyramidal shaped bone and its apex marks the ICA. Conclusion: Our anatomical model provides the means to learn the endoscopic anatomy of the PPF and may be used for the simulation of surgical techniques. An endoscopic endonasal approach provides adequate exposure to all anatomical structures within the PPF. These structures may be used as landmarks to identify and control deeper neurovascular structures. The significance is that an anatomical model facilitates learning the surgical anatomy and the acquisition of surgical skills. A dissection superficial to the vascular structures preserves the neural elements. These nerves and their bony foramina, such as the vidian nerve and V2, are critical anatomical landmarks to identify and control the ICA at the skull base.

Immunohistochemical Expression of HLA-DR in the Conjunctiva of Patients Under Topical Prostaglandin Analogs Treatment

Purpose: Subclinical inflammation may be observed in patients, using topical antiglaucomatous drugs. The objective,e of this study was to investigate inflammation in conjunctiva of glaucoma patients using prostaglandin analogs, by the detection of a immunogenetic marker (HLA-DR) and compare the effect of 3 different drugs: latanoprost, bimatoprost, and travoprost in the induction of this inflammation. Subjects and Methods: Thirty-three patients with primary open-angle glaucoma were evaluated without and with prostaglandin analogs topical therapy. Imprints of conjunctival cells were obtained, fixed on glass slides. and Prepared for immunohistochemical analysis. Results: Before the use of prostaglandin analogs, 4 of the 33 patients evaluated presented expression (of HLA-DR in the conjunctiva (mild). After 1 month oil prostaglandin analog treatment, all but 1 patient presented HLA-DR staining. HLA-DR expression of these 32 patients was scored as mild (19 patients), medium (11 patients), or intense (2 patients). The differences were statistically significant both when the presence and the increased expression of HLA-DR were considered (P<0.001). When the 3 different groups were analyzed (latanoprost, bimatoprost, and travoprost) no statistically significant difference was round (P 0.27). Conclusions: The use of prostaglandin analogs eye drops provokes, a reaction, observed by HLA-DR subclinical inflammatory expression, even after a short period of treatment, independently of the class of the prostaglandin analogs used.

A trag??dia da regi??o serrana do Rio de Janeiro em 2011: procurando respostas

O estudo de caso trata do desastre natural ocorrido na regi??o serrana do Rio de Janeiro nos dias 11 e 12 de janeiro de 2011, considerado a maior cat??strofe clim??tica e geot??cnica do pa??s, causado por fortes chuvas que provocaram enchentes e deslizamentos em sete munic??pios. O caso tem por finalidade despertar a discuss??o sobre as vari??veis e atores que influenciaram o processo de gest??o da crise, contextualizando o desastre no cen??rio da regi??o e reconstituindo, especialmente, o processo de resposta imediata dada ?? trag??dia pelos ??rg??os envolvidos. O estudo pode ser utilizado em cursos que abordem, entre outros, os seguintes t??picos: gest??o de crise e gest??o de riscos, negocia????o, comunica????o e planejamento estrat??gico

Diversidade e capacita????o em escolas de governo: mesa-redonda de pesquisa-a????o

A MRPA Diversidade e Capacita????o em Escolas de Governo reuniu especialistas e dirigentes de variadas origens para pensar a constru????o de alternativas para a inser????o dos temas diversidade, igualdade de g??nero, igualdade racial e direitos humanos nos programas de capacita????o e forma????o desenvolvidos em escolas de governo

Rede Cegonha: da concep????o ?? implanta????o

Este projeto de pesquisa realizou uma an??lise da Rede Cegonha, do Minist??rio da Sa??de, desde a sua concep????o at?? a sua implanta????o no territ??rio. Analisou os conceitos, as diretrizes, os objetivos e as intera????es dos seus elementos constitutivos, em rela????o ??s orienta????es sobre Redes de Aten????o ?? Sa??de, da Organiza????o Mundial de Sa??de (OMS) e da Organiza????o Pan-Americana de Sa??de (OPAS). Analisou, tamb??m, o processo de operacionaliza????o e de implementa????o da Rede Cegonha no ??mbito do Sistema ??nico de Sa??de. Foram utilizadas refer??ncias bibliogr??ficas nacionais e internacionais que abordam os conceitos de redes, pol??ticas p??blicas e redes de aten????o ?? sa??de e, ainda, levantamento documental como atos normativos e produ????es t??cnicas do Minist??rio da Sa??de sobre as Redes de Aten????o ?? Sa??de e a Rede Cegonha. Inicialmente foi abordado um estudo conceitual sobre redes e pol??ticas p??blicas, em seguida foi feito um detalhamento da Rede Cegonha e dos seus elementos constitutivos: a popula????o adstrita, a estrutura operacional e os modelos de aten????o ?? sa??de. Na sequencia, foram realizadas an??lises do referencial te??rico da OMS e da OPAS e a sua aplica????o na Rede Cegonha, assim como o processo de implanta????o dessa Rede no decorrer de dois anos de exist??ncia. Por fim, foram analisados os pontos fortes, os fr??geis, as oportunidades e as amea??as desta rede

Texto para discuss??o 21: burocracia, capacidade de Estado e mudan??a estrutural

Este texto visa discutir, ?? luz da literatura pertinente, o papel reservado ao Estado e, em particular, ?? burocracia, nos processos de transforma????o estrutural.Esta tem??tica, posta de lado enquanto prevaleceu o antiestatismo da vis??o neocl??ssica, vem retornando ao centro do debate acad??mico e intelectual, impulsionada pelo sucesso econ??mico dos Newly Industrialized Countries (NICs) do sudeste asi??tico. H?? fortes ind??cios da import??ncia da a????o do Estado na explica????o destes surtos de desenvolvimento.Na tentativa de elucidar estes eventos, antigas quest??es, tais como a autonomia do Estado, t??m sido alvo de crescente interesse, ao lado de conceitos como capacidade de Estado e insulamento burocr??tico. Tendo em vista as evidentes limita????es deste trabalho, n??o se pretende mais do que tangenciar os complexos problemas suscitados por esta tem??tica, a partir dos argumentos de um grupo seleto de autores

Acoplamentos: um estudo sobre a intera????o entre humanos e computadores

Este estudo investiga a intera????o dos humanos com os computadores pessoais. A partir dos inc??modos produzidos por esses encontros, na primeira parte estuda-se a produ????o da m??quina ???amig??vel??? ??? que insinua compara????es com a mente humana ??? e seus fortes efeitos nas transforma????es culturais do final do s??culo XX. Na segunda parte do trabalho, a partir de entrevistas de trabalhadores falando sobre sua aprendizagem b??sica do uso de computadores pessoais, faz-se uma an??lise das principais concep????es sobre a rela????o homem-t??cnica que est??o influenciando a intera????o humano-computador. Com isso fica evidenciado o papel do computador na crise do sujeito moderno, e afirma-se a necessidade de se pensar a subjetividade para al??m da cl??ssica divis??o sujeito-objeto, homem m??quina, natureza-artif??cio... o que aponta para a id??ia de acoplamentos flex??veis entre humanos e computadores e pede por uma no????o de aprendizagem que acolha a cria????o de problemas como seu aspecto mais rico.

Forma????o de carreiras para a gest??o p??blica contempor??nea: o caso dos especialistas em pol??ticas p??blicas e gest??o governamental

O artigo analisa o curso de forma????o inicial para a carreira de Especialista em Pol??ticas P??blicas e Gest??o Governamental (EPPGG), recentemente reformulado. Conclui-se que as mudan??as realizadas no curso, tendo em vista os problemas enfrentados em edi????es anteriores, os desafios contempor??neos da administra????o p??blica, a crescente complexidade dos problemas e as demandas da sociedade, proporcionaram respostas bastante adequadas. No entanto, persistem in??meros desafios, dentre os quais se destacam o aprimoramento do ensino de aplica????o e novas tecnologias. Ao longo das nove edi????es do curso, percebe-se que os principais objetivos do projeto inicial da carreira permaneceram, mesmo com todas as descontinuidades administrativas. Preservaram-se o perfil generalista e as policompet??ncias na forma????o do EPPGG, a fim de permitir ao profissional uma vis??o abrangente e integrada dos complexos problemas da administra????o p??blica e desenvolver a capacidade de analisar, decidir e buscar os melhores resultados em prol do cidad??o, conduzindo as rela????es e as atividades profissionais de acordo com os princ??pios da ??tica p??blica.