The Spleen CD4(+) T Cell Response to Blood-Stage Plasmodium chabaudi Malaria Develops in Two Phases Characterized by Different Properties

The pivotal role of spleen CD4(+) T cells in the development of both malaria pathogenesis and protective immunity makes necessary a profound comprehension of the mechanisms involved in their activation and regulation during Plasmodium infection. Herein, we examined in detail the behaviour of non-conventional and conventional splenic CD4(+) T cells during P. chabaudi malaria. We took advantage of the fact that a great proportion of CD4(+) T cells generated in CD1d(-/-) mice are I-A(b)-restricted (conventional cells), while their counterparts in I-Ab(-/-) mice are restricted by CD1d and other class IB major histocompatibility complex (MHC) molecules (non-conventional cells). We found that conventional CD4(+) T cells are the main protagonists of the immune response to infection, which develops in two consecutive phases concomitant with acute and chronic parasitaemias. The early phase of the conventional CD4(+) T cell response is intense and short lasting, rapidly providing large amounts of proinflammatory cytokines and helping follicular and marginal zone B cells to secrete polyclonal immunoglobulin. Both TNF-alpha and IFN-gamma production depend mostly on conventional CD4(+) T cells. IFN-gamma is produced simultaneously by non-conventional and conventional CD4(+) T cells. The early phase of the response finishes after a week of infection, with the elimination of a large proportion of CD4(+) T cells, which then gives opportunity to the development of acquired immunity. Unexpectedly, the major contribution of CD1d-restricted CD4(+) T cells occurs at the beginning of the second phase of the response, but not earlier, helping both IFN-gamma and parasite-specific antibody production. We concluded that conventional CD4(+) T cells have a central role from the onset of P. chabaudi malaria, acting in parallel with non-conventional CD4(+) T cells as a link between innate and acquired immunity. This study contributes to the understanding of malaria immunology and opens a perspective for future studies designed to decipher the molecular mechanisms behind immune responses to Plasmodium infection.

Screening of lipases for the synthesis of xylitol monoesters by chemoenzymatic esterification and the potential of microwave and ultrasound irradiations to enhance the reaction rate

Lipases from different sources, Pseudomonas fluorescens (AK lipase), Burkholderia cepacia (PS lipase), Penicillium camembertii (lipase G) and Porcine pancreas lipase (PPL), previously immobilized on epoxy SiO(2)-PVA, were screened for the synthesis of xylitol monoesters by esterification of the protected xylitol using oleic acid as acyl donor group. Among all immobilized derivatives, the highest esterification yield was achieved by P. camembertii lipase, showing to be attractive alternative to bulk chemical routes to satisfy increasing commercial demands. Further experiments were performed to determine the influence of fatty acids chain size on the reaction yield and the feasibility of using non-conventional heating systems (microwave and ultrasound irradiations) to enhance the reaction rate. (C) 2010 Elsevier B.V. All rights reserved.

A hybrid-Trefftz formulation for plane elasticity with selective enrichment of the approximations

This work is related to the so-called non-conventional finite element formulations. Essentially, a methodology for the enrichment of the initial approximation which is typical of the meshless methods and based on the clouds concept is introduced in the hybrid-Trefftz formulation for plane elasticity. The formulation presented allows for the approximation and direct enrichment of two independent fields: stresses in the domains and displacements on the boundaries of the elements. Defined by a set of elements and interior boundaries sharing a common node, the cloud notion is employed to select the enrichment support for the approximation fields. The numerical analysis performed reveals an excellent performance of the resulting formulation, characterized by the good approximation ability and a reduced computational effort. Copyright (C) 2009 John Wiley & Sons, Ltd.

Landscape unit discrimination for pedological surveys by orbital spectral response

Landscape unit discrimination for pedological surveys by orbital spectral response. The objective of tins study was compare two soil survey methods. The first was performed by methods traditionally used to distinguish landscape units and soil class discrimination. The second was based on soil class distinction through orbital spectral response. In order to establish soil characteristics and their classification, soil samples were collected at two depths in a grid system, with a distance of 500 meters between points. With these samples, physical and chemical analyses were carried out. In the sampling points, the apparent reflectance of the soil, front the orbital image, was determined and, through cluster analysis landscape units were established. In order to evaluate the resemblance reliability between the landscape units established in each method, the Kappa index was used, the value set for the confusion matrix was 0.43, indicating high quality in the comparison, showing that the non-conventional method was as close as the one carried out by photointerpretation.

Validity of (-)-[H-3]-CGP 12177A as a radioligand for the 'putative beta(4)-adrenoceptor' in rat atrium

1 We have recently suggested the existence in the heart of a 'putative beta(4)-adrenoceptor' based on the cardiostimulant effects of non-conventional partial agonists, compounds that cause cardiostimulant effects at greater concentrations than those required to block beta(1)- and Bz-adrenoceptors. We sought to obtain further evidence by establishing and validating a radioligand binding assay for this receptor with (-)-[H-3]-CGP 12177A ((-)-4-(3-tertiarybutylamino-2-hydroxypropoxy) benzimidazol-2-one) in rat atrium. We investigated (-)-[H-3]-CGP 12177A for this purpose for two reasons, because it is a nonconventional partial agonist and also because it is a hydrophilic radioligand. 2 Increasing concentrations of(-)-[H-3]-CGP 12177A, in the absence or presence of 20 mu M (-)-CGP 12177A to define non-specific binding, resulted in a biphasic saturation isotherm. Low concentrations bound to beta(1)- and beta(2)-adrenoceptors (pK(D) 9.4+/-0.1, B-max 26.9+/-3.1 fmol mg(-1) protein) and higher concentrations bound to the 'putative beta(4)-adrenoceptor' (pK(D) 7.5+/-0.1, B-max 47.7+/-4.9 fmol mg(-1) protein). In other experiments designed to exclude beta(1)- and beta(2)-adrenoceptors, (-)-[H-3]-CGP 12177A (1-200 nM) binding in the presence of 500 nM (-)-propranolol was also saturable (pK(D) 7.6+/-0.1, B-max 50.8+/-7.4 fmol mg(-1) protein). 3 The non-conventional partial agonists (-)-CGP 12177A (pK(i) 7.3+/-0.2), (+/-)-cyanopindolol (pK(i) 7.6+/-0.2), (-)-pindolol (pK(i) 6.6+/-0.1) and (+)-carazolol (pk(i), 7.2+/-0.2) and the antagonist (-)-bupranolol (pK(i) 6.6+/-0.2), all competed for (-)-[H-3]-CGP 12177A binding in the presence of 500 nM (-)-propranolol at the 'putative beta(4)-adrenoceptor', with affinities closely similar to potencies and affinities determined in organ bath studies. 4 The catecholamines competed with (-)-[H-3]-CGP 12177A at the 'putative beta(4)-adrenoceptor' in a stereoselective manner, (-)-noradrenaline (pK(iH) 6.3 +/- 0.3, pK(i), 3.5 +/- 0.1), (-)-adrenaline (pK(iH) 6.5 +/- 0.2, pK(iL) 2.9 +/- 0.1), (-)-isoprenaline (pK(iH) 6.2 +/- 0.5, pK(iL) 3.3 +/- 0.1), (+)-isoprenaline (pK(i) < 1.7), (-)-R0363 ((-)-(1-(3,4-dimethoxyphenethylamino)-3-(3,4-dihydroxyphenoxy)-2-propranol)oxalate, pK(i) 5.5 +/- 0.1). 5 The inclusion of guanosine 5-triphosphate (GTP 0.1 mM) had no effect on binding of (-)-CGP 12177A or (-)-isoprenaline to the 'putative beta(4)-adrenoceptor'. In competition binding studies, (-)-CGP 12177A competed with (-)-[H-3]-CGP 12177A for one receptor state in the absence (pK(i) 7.3 +/- 0.2) or presence of GTP (pK(i) 7.3 +/- 0.2). (-)-Isoprenaline competed with (-)-[H-3]-CGP 12177A for two states in the absence (pK(iH) 6.6 +/- 0.3, pK(iL) 3.5 +/- 0.1; % H 25 +/- 7) or presence of GTP (pK(iH) 6.2 +/- 0.5, pK(iL) 3.4 +/- 0.1; % H 37 +/- 6). In contrast, at beta(1)-adrenoceptors, GTP stabilized the low affinity state of the receptor for (-)-isoprenaline. 6 The specificity of binding to the 'putative beta(4)-adrenoceptor' was tested with compounds active at other receptors. High concentrations of the beta(4)-adrenoceptor agonists, BRL 37344 ((RR + SS)[4-[2-[[2-(3-chlorophenyl)-2-hydroxy -ethyl]amino]propyl]phenoxy]acetic acid, 6 mu M), SR 58611A (ethyl((7S)-7-[(2R)-2-(3-chlorophenyl)-2-hydroxyethylamino]-5,6,7,8-tetrahydronaphtyl-2-yloxy) acetate hydrochloride, 6 mu M), ZD 2079 ((+/-)-1-phenyl-2-(2-4-carboxymethylphenoxy)-ethylamino)ethan-1-ol, 60 mu M), CL 316243 (disodium (R,R)-5-[2-[2-(3-chlorophenyl)-2-hydroxyethyl-amino]propyl]- 1,3-benzodioxole-2,2-dicarboxylate, 60 mu M) and antagonist SR 59230A (3-(2-ethylphenoxy)-1-[(1S)-1,2,3,4-tetrahydronaphth-1-ylamino]-2S-2-propanol oxalate, 6 mu M) caused less than 22% inhibition of (-)-[H-3]-CGP 12177A binding in the presence of 500 nM (-)-propranolol. Histamine (1 mM), atropine (1 mu M), phentolamine (10 mu M), 5-HT(100 mu M) and the 5-HT4 receptor antagonist SE 207710 ((1-butyl-4-piperidinyl)-methyl 8-amino-7-iodo-1 ,4-benzodioxan-5-carboxylate, 10 nM) caused less than 26% inhibition of binding. 7 Non-conventional partial agonists, the antagonist (-)-bupranolol and catecholamines all competed for (-)-[H-3]-CGP 12177A binding in the absence of (-)-propranolol at beta(1)-adrenoceptors, with affinities (pK(i)) ranging from 1.6-3.6 log orders greater than at the 'putative beta(4)-adrenoceptor'. 8 We have established and validated a radioligand binding assay in rat atrium for the 'putative beta(4)-adrenoceptor' which is distinct from beta(1)-, beta(2)- and beta(3)-adrenoceptors. The stereoselective interaction with the catecholamines provides further support for the classification of the receptor as 'putative beta(4)-adrenoceptor'.

Morphine has a dual concentration-dependent effect on K+-evoked substance P release from rat peripheral airways

Our previous investigations of possible lung mechanisms underlying the effectiveness of nebulized morphine for the relief of dyspnoea, have shown a high density of non-conventional opioid binding sites in rat airways with similar binding characteristics (opioid alkaloid-sensitive, opioid peptide-insensitive) to that of putative mu(3)-opioid receptors on immune cells. To investigate whether these lung opioid binding sites are functional receptors, this study was designed to determine (using superfusion) whether morphine modulates the K+-evoked release of the pro-inflammatory neuropeptide, substance P (SP), from rat peripheral airways. Importantly, K+-evoked SP release was Ca2+-dependent, consistent with vesicular release. Submicromolar concentrations of morphine (1 and 200 nM) inhibited K+-evoked SP release from rat peripheral airways in a naloxone (1 mu M) reversible manner. By contrast, 1 mu M morphine enhanced K+-evoked SP release and this effect was not reversed by 1 mu M naloxone. However, 100 mu M naloxone not only antagonized the facilitatory effect of 1 mu M morphine on K+-evoked SP release from rat peripheral airways but it inhibited release to a similar extent as 200 nM morphine. It is possible that these latter effects are mediated by non-conventional opioid receptors located on mast cells, activation of which causes naloxone-reversible histamine release that in turn augments the release of SP from sensory nerve terminals in the peripheral airways. Clearly, further studies are required to investigate this possibility. (C) 1997 Academic Press Limited.

Modulation of human cardiac function through 4 beta-adrenoceptor populations

In human heart there is now evidence for the involvement of four beta-adrenoceptor populations, three identical to the recombinant beta(1)-, beta(2)- and beta(3)-adrenoceptors, and a fourth as yet uncloned putative beta-adrenoceptor population, which we designate provisionally as the cardiac putative beta(4)-adrenoceptor. This review described novel features of beta-adrenoceptors as modulators of cardiac systolic and diastolic function. We also discuss evidence for modulation by unoccupied beta(1)- and beta(2)-adrenoceptors. Human cardiac and recombinant beta(1)- and beta(2)-adrenoceptors are both mainly coupled to adenylyl cyclase through Gs protein, the latter more tightly than the former. Activation of both human beta(1)- and beta(2)-adrenoceptors not only increases cardiac force during systole but also hastens relaxation through cyclic AMP-dependent phosphorylation of phospholamban and troponin I, thereby facilitating diastolic function. Furthermore, both beta(1) and beta(2)-adrenoceptors can mediate experimental arrhythmias in human cardiac preparations elicited by noradrenaline and adrenaline. Human ventricular beta(3)-adrenoceptors appear to be coupled to a pertussis toxin-sensitive protein (Gi?). beta(3)-Adrenoceptor-selective agonists shorten the action potential and cause cardiodepression, suggesting direct coupling of a Gi protein to a K+ channel. In a variety of species, including man, cardiac putative beta(4)-adrenoceptors mediate cardiostimulant effects of non-conventional partial agonists, i.e. high affinity beta(1)- and beta(2)-adrenoceptor blockers that cause agonist effects at concentrations considerably higher than those that block these receptors. Putative beta(4)-adrenoceptors appear to be coupled positively to a cyclic AMP-dependent cascade and can undergo some desensitisation.

Abolition of (-)-CGP 12177-evoked cardiostimulation in double beta(1)/beta(2)-adrenoceptor knockout mice. Obligatory role of beta(1)-adrenoceptors for putative beta 4-adrenoceptor pharmacology

Some beta (1)- and beta (2)-adrenoceptor-blocking agents, such as (-)-CGP 12177, cause cardiostimulant effects at concentrations considerably higher than those that antagonise the effects of catecholamines. The cardiostimulant effects of these non-conventional partial agonists are relatively resistant to blockade by (-)-propranolol and have been proposed to be mediated through putative beta (4)-adrenoceptors or through atypical states of either beta (1)- or beta (2)-adrenoceptors. We investigated the effects of (-)-CGP 12177 on sinoatrial rate and left atrial contractile force as well as the ventricular binding of (-)-[H-3]CGP 12177 in tissues from wild-type, beta (2)-adrenoceptor knockout and beta (1)/beta (2)-adrenoceptor double knockout mice. The cardiostimulant effects of (-)-CGP 12177 were present in wildtype and beta (2)-adrenoceptor knockout mice but were absent in beta (1)/beta (2)-adrenoceptor double knockout mice. Thus, the presence of beta (1)-adrenoceptors is obligatory for the cardiostimulant effects of (-)-CGP 12177. It appears therefore that an atypical state of the beta (1)-adrenoceptor contributes to the mediation of the cardiostimulant effects induced by non-conventional partial agonists. Ventricular beta (1)- and beta (2)-adrenoceptors, labelled in wild-type with a K(D)similar to0.5 nmol/l (similar to 16 fmol/mg protein), were absent in beta (1)/beta (2)-adrenoceptor double knockout mice. However, a high density binding site (similar to 154-391 fmol/mg protein) that did not saturate completely (K(D)similar to 80-200 nM) was labelled by (-)-[H-3]CGP 12177 in the three groups of mice, being distinct from beta (1)- and beta (2)-adrenoceptors, as well as from the site mediating the agonist effects of(-)-CGP 12177.

1-Ethynyl-2-isopropoxy-3-methoxybenzene

The crystal structure of the title compound, C12H14O2, has been determined at 150 K. Significant intermolecular non-conventional C-H...O interactions involving the terminal acetylinic H atom are observed, which result in a zigzag hydrogen-bonded chain in the b direction.

Programa de Técnica de Dança Clássica : do pré-primário ao 2ª ciclo de escolaridade do Ensino Não Vocacional

Relatório Final de Projecto Pedagógico apresentado à Escola Superior de Dança, com vista à obtenção do grau de Mestre em Metodologias do Ensino da Dança

O teatro. espaço de encontro de uma comunidade

Dissertação apresentada à Escola Superior de Educação de Lisboa para obtenção de grau de mestre em Educação Artística - Especialização em Teatro na Educação

Perspetiva da comunidade docente sobre a relação entre a autoavaliação da escola e o seu projeto curricular: um estudo de caso numa escola básica e secundária

Dissertação de Mestrado, Educação (Concepção e Desenvolvimento de Projectos Educativos), 21 de Janeiro de 2013, Universidade dos Açores.

Simulação de processos de adsorção recorrendo ao Aspen Chromatography

As operações de separação por adsorção têm vindo a ganhar importância nos últimos anos, especialmente com o desenvolvimento de técnicas de simulação de leitos móveis em colunas, tal como a cromatografia de Leito Móvel Simulado (Simulated Moving Bed, SMB). Esta tecnologia foi desenvolvida no início dos anos 60 como método alternativo ao processo de Leito Móvel Verdadeiro (True Moving Bed, TMB), de modo a resolver vários dos problemas associados ao movimento da fase sólida, usuais nestes métodos de separação cromatográficos de contracorrente. A tecnologia de SMB tem sido amplamente utilizada em escala industrial principalmente nas indústrias petroquímica e de transformação de açúcares e, mais recentemente, na indústria farmacêutica e de química fina. Nas últimas décadas, o crescente interesse na tecnologia de SMB, fruto do alto rendimento e eficiente consumo de solvente, levou à formulação de diferentes modos de operação, ditos não convencionais, que conseguem unidades mais flexíveis, capazes de aumentar o desempenho de separação e alargar ainda mais a gama de aplicação da tecnologia. Um dos exemplos mais estudados e implementados é o caso do processo Varicol, no qual se procede a um movimento assíncrono de portas. Neste âmbito, o presente trabalho foca-se na simulação, análise e avaliação da tecnologia de SMB para dois casos de separação distintos: a separação de uma mistura de frutose-glucose e a separação de uma mistura racémica de pindolol. Para ambos os casos foram considerados e comparados dois modos de operação da unidade de SMB: o modo convencional e o modo Varicol. Desta forma, foi realizada a implementação e simulação de ambos os casos de separação no simulador de processos Aspen Chromatography, mediante a utilização de duas unidades de SMB distintas (SMB convencional e SMB Varicol). Para a separação da mistura frutose-glucose, no quediz respeito à modelização da unidade de SMB convencional, foram utilizadas duas abordagens: a de um leito móvel verdadeiro (modelo TMB) e a de um leito móvel simulado real (modelo SMB). Para a separação da mistura racémica de pindolol foi considerada apenas a modelização pelo modelo SMB. No caso da separação da mistura frutose-glucose, procedeu-se ainda à otimização de ambas as unidades de SMB convencional e Varicol, com o intuito do aumento das suas produtividades. A otimização foi realizada mediante a aplicação de um procedimento de planeamento experimental, onde as experiências foram planeadas, conduzidas e posteriormente analisadas através da análise de variância (ANOVA). A análise estatística permitiu selecionar os níveis dos fatores de controlo de modo a obter melhores resultados para ambas as unidades de SMB.

Teatro e igualdade de género : uma intervenção social e artística : relatório de estágio

O presente Relatório de Estágio tem como objecto de pesquisa a concepção e monitorização de duas Oficinas de Teatro dirigidas a dois grupos de mulheres constituídos, respectivamente, por estudantes universitárias e desempregadas. O estágio realizou-se entre Novembro/2013 e Junho/2014 na Quarta Parede - Associação de Artes Performativas da Covilhã e inseriu-se nos Empowerment Labs, laboratórios formativos que cruzam artes performativas e ciências sociais na reflexão e intervenção sobre a igualdade de género com foco no desemprego feminino. Este relatório expõe os três momentos do processo do estágio: pesquisa de referenciais teórico-práticos, concepção e monitorização das Oficinas de Teatro e reflexão a partir da prática laboratorial. Na pesquisa de referenciais, essencial para delinear a metodologia operacional e o programa de conteúdos, explorei dimensões como o feminismo e a igualdade de género, e procurei compreender de que forma o empowerment, a pedagogia de Paulo Freire e as metodologias do teatro aplicado serviam os objectivos do meu trabalho. A realização das oficinas foi o momento de experimentar as metodologias e o programa delineado. Tendo a igualdade de género como temática unificadora, o empowerment através da arte como objectivo maior e o teatro aplicado como base metodológica, as oficinas inserem-se nos processos de educação não-formal aplicados ao incremento de recursos intelectuais, emocionais, sociais, expressivos e criativos e, neste caso específico, à ampliação da consciência de género. Neste sentido, as oficinas desenvolveram uma abordagem metodológica processual, participativa e multidisciplinar, orientada para a pesquisa performativa, primeiro de uma dramaturgia individual, depois de uma dramaturgia do colectivo e, por fim, de uma dramaturgia orientada para a igualdade de género. O conceito de “dramaturgia” surge aqui no sentido metodológico do pachwork (trabalho com retalhos) e do sampling (recolha e transformação de materiais), relevante sobretudo na última fase, dedicada à construção colectiva de um exercício performativo apresentado publicamente.

Partitura de luz: a representação criativa como perspectiva educacional

Este projecto de investigação teve como objectivo avaliar - através de uma série de workshops orientados pela mestranda no Centro Cultural de Belém em Abril de 2012 - o impacto da utilização de notação musical não-convencional num contexto não escolar. Traçando possíveis paralelos com o ensino especializado da música, propuseram-se metodologias de aprendizagem que permitissem a introdução da notação, de forma inovadora, no referido contexto escolar. O processo de investigação baseou-se em observação directa, na análise dos questionários preenchidos pelos participantes dos workshops e na observação e análise das gravações em vídeo que documentaram o processo artístico, pedagógico e de investigação. Numa sala escura, 6 retroprojectores projectaram uma Partitura de Luz. Entre crianças e adultos, 120 participantes (não-músicos) criaram empiricamente as suas composições, moldando o som em função da forma e a forma em função do som. O resultado foi compensador: a criação de condições favoráveis ao desenvolvimento máximo da expressão criativa individual ou colectiva dos participantes - através da utilização irrestrita de símbolos, imagens, objectos e matérias - culminou num efectivo estabelecimento de correspondência musical, a partir de recursos vocais. Este projecto, “Partitura de Luz”, foi uma oportunidade de relacionar a vertente artística – musical, plástica e gráfica - com a vertente humana: pedagógica e social.