Local and systemic delivery of a novel group of inhibitors of transglutaminase enzyme: A potential approach for treating of catheter-related complications and liver fibrosis

The present thesis investigates targeted (locally and systemically) delivery of a novel group of inhibitors of enzyme transglutaminases (TGs). TGs are a widely distributed group of enzymes that catalyse the formation of isopeptide bonds between the y-carboxamide group of protein-bound glutamines and the a-amino group of protein-bound lysines or polyamines. The first group of the novel inhibitors tested were the tluorescently labelled inhibitors of Factor XIIIa (FXIIIa). These small, non-toxic inhibitors have the potential to prevent stabilisation of thrombi by FXIIIa and consequently increase the natural rate of thrombolysis, in addition it reduces staphylococcal colonisation of catheters by inhibiting their FXIIIa¬mediated cross-linking to blood clot proteins on the central venous catheter (CVCs) surface. The aim of this work was to incorporate the FXIIIa inhibitor either within coating of polyurethane (PU) catheters or to integrate it into silicone catheters, so as to reduce the incidence of thrombotic occlusion and associated bacterial infection in CVCs. The initial work focused on the incorporation of FXIIIa inhibitors within polymeric coatings of PU catheters. After defining the key characteristics desired for an effective polymeric-coating, polyvinylpyrrolidone (PVP), poly(lactic-co-glycolic acid) (PLGA) or their combination were studies as polymers of choice for coating of the catheters_ The coating was conducted by dip-coating method in a polymer solution containing the inhibitor. Upon incubation of the inhibitor-and polymer-coated strips in buffer, PVP was dissolved instantly, generating fast and significant drug release, whilst PLGA did not dissolve, yielding a slow and an insufficient amount of drug release. Nevertheless, the drug release profile was enhanced upon employing a blend solution of PVP and PLGA. The second part of the study was to incorporate the FXIIIa inhibitor into a silicone elastomer; results demonstrated that FXIIIa inhibitor can be incorporated and released from silicone by using citric acid (CA) and sodium bicarbonate (SB) as additives and the drug release rate can be controlled by the amount of incorporated additives in the silicone matrix. Furthermore, it was deemed that the inhibitor was still biologically active subsequent to being released from the silicone elastomer strips. Morphological analysis confirmed the formation of channels and cracks inside the specimens upon the addition of CA and SB. Nevertheless, the tensile strength, in addition to Young's modulus of silicone elastomer strips, decreased constantly with an increasing amount of amalgamated CA/ SB in the formulations. According to our results, incorporation of FXIIIa inhibitor into catheters and other medical implant devices could offer new perspectives in preventing bio-material associated infections and thrombosis. The use of tissue transglutaminase (T02) inhibitor for treating of liver fibrosis was also investigated. Liver fibrosis is characterized by increased synthesis and decreased degradation of the extracellular matrix (ECM). Transglutaminase-mediated covalent cross-linking is involved in the stabilization of ECM in human liver fibrosis. Thus, TG2 inhibitors may be used to counteract the decreased degradation of the ECM. The potential of a liposome based drug delivery system for site specific delivery of the fluorescent TG2 inhibitor into the liver was investigated; results indicated that the TG2 inhibitor can be successfully integrated into liposomes and delivered to the liver, therefore demonstrating that liposomes can be employed for site-specific delivery of TG2 inhibitors into the liver and TG2 inhibitor incorporating liposomes could offer a new approach in treating liver fibrosis and its end stage disease cirrhosis.

Planning of schools as communities

The purpose of this research paper was to conduct a literature review to explore both the physical and historical aspects of schools and their communities in order to ascertain the nature of the disconnect that exists today between schools and communities. This paper reviews the purpose of the public school, from the origin of the one-room schoolhouse to today's urban schools; the impact of immigration, segregation, technology and industrialization on the public school and the forces behind public school reform. The reform processes and the programs developed to stimulate the public school including the transformation of the learning environment in the communities they serve was discussed. The research indicates that the return to a community school with small learning environments that involves communities and residents in the planning of schools as communities, is the most promising form of public school reform.

The many voices of Piedmont: language, identity and alterity in the works of modern and contemporary Piedmontese writers

The aim of this project is to carry out a linguistic analysis of a group of modern and contemporary narratives written by authors from the same Italian region: Piedmont. The novels and short stories examined stand out for the intriguing ways in which they move between a variety of idioms – Italian, Piedmontese dialects, English and pastiches, with some rare excursions into French. A sociolinguistic study and an overview of political changes that Piedmont underwent from the sixteenth to the twenty-first centuries are provided, with the purpose of outlining the region’s sociogeographical and historical background which can be seen to have fostered multilingualism in a group of writers. With the support of linguistic studies and philosophical theories on the relation between identity, alterity and language (such as Edwards’s Language and Identity and Bakhtin’s reflections on language), I then elucidate the presence of diverse linguistic varieties in selected narratives by Cesare Pavese, Beppe Fenoglio, Primo Levi, Nanni Balestrini, Fruttero & Lucentini, Benito Mazzi and Younis Tawfik. In other words, my purpose is to explain the reasons for multilingualism in each writer, as well as to underscore the ideological positions which lie behind the linguistic strategies of the authors. With this study I attempt to fill a gap and cast new light on Piedmontese literature. Although some critical studies on the use of dialect or English exist on individual authors and works (e.g. Meddemmem on Fenoglio’s use of English and Beccaria on Pavese’s inclusion of Piedmontese dialect), and some important contributions to the history of Piedmontese literature have appeared in print, to date no current, systematic study that compares different Piedmontese writers under the language/identity theme has been published. The study concludes with a summary of the evolution of plurilingualism in Piedmont and highlights the common trends in the use of multiple linguistic varieties as tools for both social demarcation and an opening up to alternative, marginalised andforeign cultures.

Differences In Metabolic Profile In the Testes of Streptozotocin Induced Diabetes Mellitus Mice As Detected By 1H NMR

Contrary to the traditional view, recent studies suggest that diabetes mellitus has an adverse influence on male reproductive function. Our aim was to determine the affect of diabetes on the testicular environment by identifying and then assessing perturbations in small molecule metabolites. Testes were obtained from control and streptozotocin induced diabetic C57BL/6 mice, two, four and eight weeks post treatment. Diabetic status was confirmed by HbA1c, non fasting blood glucose, physiological condition and body weight. Protein free, low molecular weight, water soluble extracts were assessed using 1H NMR spectroscopy. Principal Component Analysis of the derived profiles was used to classify any variations and specific metabolites were identified based on their spectral pattern. Characteristic metabolite profiles were identified for control and diabetic animals with the most distinctive being from mice with the greatest physical deterioration and loss of bodyweight. Eight streptozotocin treated animals did not develop diabetes and displayed profiles similar to controls. Diabetic mice had decreases in creatine, choline and carnitine and increases in lactate, alanine and myo-inositol. Betaine levels were found to be increased in the majority of diabetic mice but decreased in two animals with severe loss of body weight and physical condition. The association between perturbations in a number of small molecule metabolites known to be influential in sperm function, with diabetic status and physiological condition, adds further impetus to the proposal that diabetes influences important spermatogenic pathways and mechanisms in a subtle and previously unrecognised manner.

Metabolic profile changes in the testes of mice with streptozotocin-induced type 1 diabetes mellitus

Contrary to the traditional view, recent studies suggest that diabetes mellitus has an adverse influence on male reproductive function. Our aim was to determine the effect of diabetes on the testicular environment by identifying and then assessing perturbations in small molecule metabolites. Testes were obtained from control and streptozotocin-induced diabetic C57BL/6 mice, 2, 4 and 8 weeks post-treatment. Diabetic status was confirmed by glycated haemoglobin, non-fasting blood glucose, physiological condition and body weight. A novel extraction procedure was utilized to obtain protein free, low-molecular weight, water soluble extracts which were then assessed using H-1 nuclear magnetic resonance spectroscopy. Principal component analysis of the derived profiles was used to classify any variations, and specific metabolites were identified based on their spectral pattern. Characteristic metabolite profiles were identified for control and type 1 diabetic animals with the most distinctive being from mice with the largest physical deterioration and loss of body weight. Eight streptozotocin-treated animals did not develop diabetes and displayed profiles similar to controls. Diabetic mice had decreases in creatine, choline and carnitine and increases in lactate, alanine and myo-inositol. Betaine levels were found to be increased in the majority of diabetic mice but decreased in a few animals with severe loss of body weight and physical condition. The association between perturbations in a number of small molecule metabolites known to be influential in sperm function, with diabetic status and physiological condition, adds further impetus to the proposal that diabetes influences important spermatogenic pathways and mechanisms in a subtle and previously unrecognized manner.

In-vitro investigation of out-of-field cell survival following the delivery of conformal, intensity-modulated radiation therapy (IMRT) and volumetric modulated arc therapy (VMAT) plans

The aim of this work is to determine the out-of-field survival of cells irradiated with either the primary field or scattered radiation in the presence and absence of intercellular communication following delivery of conformal, IMRT and VMAT treatment plans. Single beam, conformal, IMRT and VMAT plans were created to deliver 3 Gy to half the area of a T80 flask containing either DU-145 or AGO-1522 cells allowing intercellular communication between the in-and out-of-field cell populations. The same plans were delivered to a similar custom made phantom used to hold two T25 culture flasks, one flask in-field and one out-of-field to allow comparison of cell survival responses when intercellular communication is physically inhibited. Plans were created for the delivery of 8 Gy to the more radio-resistant DU-145 cells only in the presence and absence of intercellular communication. Cell survival was determined by clonogenic assay. In both cell lines, the out-of-field survival was not statistically different between delivery techniques for either cell line or dose. There was however, a statistically significant difference between survival out-of-field when intercellular communication was intact (single T80 culture flask) or inhibited (multiple T25 culture flasks) to in-field for all plans. No statistically significant difference was observed in-field with or without cellular communication to out-of-field for all plans. These data demonstrate out-of-field effects as important determinants of cell survival following exposure to modulated irradiation fields when cellular communication between differentially irradiated cell populations is present. This data is further evidence that refinement of existing radiobiological models to include indirect cell killing effects is required.

The role of sexual steroids in the modulation of growth hormone (GH) secretion in humans

Sex steroids contribute to modulate GH secretion in man. However, both the exact locus and mechanism by which their actions are exerted still remain not clearly understood. We undertook a number of studies designed to ascertain: (1) whether or not sudden or chronic changes in circulating gonadal steroids may affect GH secretion in normal adults; and (2) the reason(s) for gender-related dimorphic pattern of GH release. The pituitary reserve of GH, as evaluated by means of a GHRH challenge, was similar in women with anorexia nervosa and in normally menstruating women. Estrogenic receptor blockade with tamoxifen (TMX) did not significantly change GHRH-induced GH response in these normal women. Therefore, acute or chronic hypoestrogenism apparently had no important effects at level of somatotrophs. In another group of normal women we tested the possibility that changes in circulating estrogens might induce changes in the hypothalamic-somatotroph rhythm (HSR). GHRH challenges were performed throughout a menstrual cycle, and again after having achieved functional ovarian blockade with a GnRH agonist treatment. Short-term ovarian blockade did not significantly affect the parameters of GH response to GHRH, although it was accompanied by an increase in the number of women ina refractory HSR phase at testing. This suggested a low potentiating effect on the basic pattern of somatostatin (SS) release occurring as a consequence of the decrease in circulating estrogens. In normal men, neither the GH response to GHRH nor the HSR were affected by functional testicular blockade (after GnRH agonist treatment). However, the administration of testosterone enanthate (250 mg) to another group of men increased both the GHRH-induced GH release and the number of subjects in a spontaneous secretory HSR phase at testing; these were reversed by estrogenic receptor blockade with TMS. In another group of normal men, the fraction of GH secreted in pulses (FGHP) during a nocturnal sampling period was significantly decreased by testicular blockade. Other parameters of GH secretion, such as the number of GH pulses and their mean amplitude (A), and the mean plasma GH concentration (MCGH), showed a slight, although not significant, decrease following the lack of androgens. The administration of testosterone enanthate (500 mg) reversed these parameters to values similar to those in the basal study. Interestingly, when tamoxifen was given after testosterone enanthate, A, MCGH and FGHP increased to values significantly higher than in any other experimental condition in that study. In all, these data suggest that 17ß-estradiol may participate in GH modulation by inhibiting the hypothalamic release of somatostatin, while testosterone stimulates it. The results obtained after estrogenic receptor blockade appear to indicate that the effect of testosterone in such a modulation is dependent on its aromatization to 17ß-estradiol. The differential levels of this steroid in both sexes might account for the sexual dimorphic pattern of GH secretion. From other data in the literature, obtained in rats, and our preliminary data in children with constitutional delay of growth and puberty, it is tempting to speculate that the effect of 17ß-estradiol may be exerted by modifying the functional activity of a-2 adrenergic pathways involved in the negative modulation of SS release.

In vitro viability of choroidal melanomata following pre-enucleation irradiation

In a group of eighteen patients with uveal melanomas, seven underwent low-dose pre-enucleation irradiation of approximately 2000 cGy. All the tumours were propagated in tissue culture and the growth characteristics of tumour cells from irradiated eyes were compared with tumour cells from non-irradiated eyes. Cultures were observed with phase-contrast microscopy, and radioactive thymidine labelling was used to study cell turnover. Although tissue samples from peripheral areas of irradiated tumours produced a mixture of viable and non-viable cells, with reduced ability to attach to substrate, central regions of irradiated tumours contained viable cells which propagated freely in tissue culture.

Dorsal and ventral stream mediated visual processing in genetic subtypes of Prader-Willi syndrome

Previous work has suggested that there are specific deficits in dorsal stream processing in a variety of developmental disorders. Prader-Willi syndrome (PWS) is associated with two main genetic subtypes, deletion and disomy. Relative strengths in visual processing are shown in PWS, although these strengths may be specific to the deletion subtype. We investigated visual processing in PWS using an adapted Simon task which contrasted location (dorsal stream) and shape identity (ventral stream) tasks. Compared to a group of typically developing children, children with PWS deletion showed a greater degree of impairment in the dorsal stream task than in the ventral stream task, a pattern similar to that shown in a group of boys with Fragile-X syndrome. When matched on a measure of non-verbal ability, children with PWS disomy showed the opposite pattern with better performance in the location compared to the shape task, although these task performance asymmetries may have been linked to executive control processes. It is proposed that children with PWS deletion show a relative strength in visual processing in the ventral stream along with a specific deficit in dorsal stream processing. In contrast, children with PWS disomy show neither effect. (C) 2009 Published by Elsevier Ltd.

Avaliação do desempenho docente e o professor titular: um estudo de caso numa perspetiva organizacional

A avaliação do desempenho docente tem vindo a adquirir uma centralidade crescente nos contextos educativos. Para além de razões mais abrangentes que têm a ver com a importância gradual que a avaliação adquire nas políticas educativas, considera-se que a avaliação do desempenho docente contribui para a melhoria da qualidade do trabalho docente, das aprendizagens e dos resultados escolares dos alunos. As alterações do Estatuto da Carreira Docente introduziram, a partir de 2007, a par com um novo e mais exigente sistema de avaliação, a divisão da carreira em duas categorias – professor e professor titular. Os professores titulares passaram a desempenhar funções de avaliação e coordenação dos seus colegas. Estas mudanças trouxeram alterações significativas na carreira, na profissão e no trabalho docente. O propósito deste estudo foi pesquisar, do ponto de vista da análise organizacional, o modo como foi percecionado pelos docentes o processo de avaliação do desempenho (relativo ao ciclo 2007-2009), centrando-nos, em particular, no papel desempenhado pelos professores titulares num modelo de avaliação por pares e num quadro legal de verticalização da carreira docente. Desenvolvemos este estudo num agrupamento de escolas da zona centro do território continental português seguindo uma metodologia de estudo de caso. Como principais conclusões, e tendo em conta as lógicas organizacionais de que partimos, ressalta-se: i) ao nível da lógica burocrática, a burocratização e morosidade do concurso dos professores titulares, a complexidade dos procedimentos conducentes à implementação do sistema de avaliação e o formalismo na construção dos respetivos instrumentos; ii) dentro da lógica conflitual, as tensões decorrentes da discordância com a divisão da carreira entre professor e professor titular, a atribuição aos professores titulares da função avaliativa, não tendo estes sido reconhecidos pelos seus pares com legitimidade para os avaliar, e a existência de conflitos entre os diversos intervenientes; iii) na lógica artificial, o facto de este sistema de avaliação não ter implicado alterações significativas nas práticas, tendo em conta a exígua adesão dos professores à avaliação da componente cientifico-pedagógica, a ritualização de processos e o restringir-se ao cumprimento dos requisitos mínimos.

11-ketotestosterone inhibits the alternative mating tactic in sneaker males of the peacock blenny, Salaria pavo

In the peacock blenny, Salaria pavo, a species with courtship sex-role reversal, smaller, younger males mimic the courtship behavior and the nuptial coloration of females in order to get access to nests during spawning and to parasitize egg fertilization from nest-holder males. Later in their life, sneakers transform both morphologically and behaviorally into nest-holder males. In the present paper we investigate the activational role of 11-ketotestosterone (KT), the most potent androgen in most teleost species, to promote the switch between tactics in sneaker males of S. pavo. Sneakers were implanted either with KT or with control (i.e. castor oil) silastic implants. A week after implantation they were subjected to a set of behavioral tests and morphometric measurements. KT treatment promoted the differentiation of secondary sex characters, such as the anal glands, and inhibited the expression of female courtship behavior. KT-treated sneakers also showed a trend toward less frequent display of female nuptial coloration. There was no effect of KT treatment on the expression of typical nest-holder male behavior. Finally, there was no effect of KT treatment on the number or soma size of arginine vasotocin neurons in the preoptic area, which are often associated with the expression of vertebrate sexual behavior. Thus, KT seems to play a key role in mating tactic switching by inhibiting the expression of female courtship behavior and by promoting the development of male displaying traits (e.g. anal glands). The lack of a KT effect on behavior typical of nest-holding males and vasotocinergic preoptic neurons suggests that a longer time frame or other endocrine/social signals are needed for the initiation of these traits in males that are switching tactics.

Avaliação de desempenho docente: efeitos no desenvolvimento profissional.

Dissertação apresentada à Escola Superior de Educação de Lisboa para obtenção de grau de mestre em Ciências da Educação - Especialização Supervisão em Educação

A comunicação entre o diretor e os professores no agrupamento X

A comunicação é essencial nas relações interpessoais, quer ao nível das organizações em geral, quer na organização escolar em particular. As funções de administração, como planeamento, organização, liderança e controle, são levadas a cabo com sucesso, dentro de uma organização quando apoiadas num processo de comunicação eficaz. O objectivo deste estudo foi procurar soluções para a ineficácia relativa do processo de comunicação entre o Diretor e os professores no Agrupamento X. Metodologicamente desenvolveu-se um estudo de caso, de natureza qualitativa, onde se recorreu a instrumentos de recolha de dados como entrevistas e ‘focus group’. Desta forma identificou-se e descreveu-se o processo de comunicação adotado no Agrupamento X entre o Diretor e os professores, bem como se recolheram as opiniões dos docentes e do Diretor sobre a comunicação e a gestão da informação no Agrupamento. Este estudo permitiu reconhecer que a fraca internalização de alguns conceitos e mecanismos do processo comunicacional por parte dos atores contribui para ineficácia relativa deste processo. Na criação do plano de ação retomámos e valorizamos a ideia de que um modelo de comunicação assente nas principais funções da comunicação poderá ser a chave para o maior envolvimento dos docentes na consecução dos objetivos do Projeto Educativo do Agrupamento, ou seja, é dada a resposta à pergunta partida deste projeto – como melhorar o processo de comunicação entre o Diretor e os professores no Agrupamento X?

Autoavaliação das organizações escolares: articulação da equipa de autoavaliação com as lideranças intermédias

O presente trabalho de projeto resulta de um estudo na área da Avaliação Institucional, realizado num agrupamento de escolas com educação pré-escolar e ensinos básico e secundário. O objetivo principal relacionou-se com a construção de um Plano de Ação, que promova o desenvolvimento de uma cultura de avaliação, procurando envolver todos os ‘atores’ educativos no processo de mudança e melhoria do agrupamento, com principal incidência na articulação da Equipa de Autoavaliação do Agrupamento, com as Lideranças Intermédias, de forma a implementar o processo de Autoavaliação do Agrupamento. A avaliação das escolas deve basear-se em dispositivos simples e exequíveis e nesse sentido é importante que a avaliação respeite critérios de pertinência, de coerência, de eficácia, de eficiência e de oportunidade. As Equipas de Autoavaliação, em articulação com as Lideranças Intermédias, assumem cada vez mais importância, nas organizações escolares, podendo ser potenciadoras da melhoria da eficácia e eficiência das organizações escolares. Um dos instrumentos de recolha de dados foi o inquérito por questionário, aplicado a 89 professores e educadores, que representam 51,2 % dos docentes do agrupamento em estudo. Outro instrumento de recolha de dados foi o focus group, constituído por 11 professores, dos departamentos curriculares existentes no agrupamento. Os resultados mostraram que o processo de autoavaliação é já uma prática comum no agrupamento, com uma certa consolidação. No entanto, ao nível da estrutura que lidera, apesar de integrar representantes de toda a comunidade educativa, ainda se verifica uma participação muito centrada nos docentes. Esta estrutura deve tornar-se mais abrangente, de forma a permitir a incorporação de outros pontos de vista. Os resultados mostraram ainda, diferentes tipos de abordagem aos assuntos relacionados com a autoavaliação, nos distintos departamentos curriculares. O incentivo das Lideranças Intermédias e a sua implicação na tarefa de motivar os professores para a participação no processo, a articulação com a Equipa de Autoavaliação e a comunicação serão determinantes para a melhoria da eficácia e eficiência da organização. No plano de ação, é apresentado um conjunto de propostas, que pretendem colmatar estas debilidades.

O fio de Ariadne : a formação contínua do PNEP e desenvolvimento da competência da leitura no 1.º CEB

Afirmando-se como uma necessidade premente, a qualidade da formação contínua de professores tem evoluído nos últimos anos. Esta urgência surge a partir das exigências que cada vez mais se sentem, quer a nível nacional, quer internacional, de um ensino e aprendizagem que acompanhe as mudanças sociais, tecnológicas e multiculturais. O Programa Nacional de Ensino do Português (PNEP), que surgiu pela necessidade de aumentar os níveis de literacia dos alunos portugueses, foi o objeto de estudo desta investigação. Nesta perspetiva, pretendeu-se identificar de que forma a formação de professores se articula com as dinâmicas da formação centrada nas escolas, no âmbito do PNEP, no desenvolvimento de competências de leitura dos alunos do 1.º CEB. Embora a aprendizagem do português padrão passe pelo ensino e aprendizagem de vários domínios, focalizou-se o estudo na relação entre a atitude reflexiva do professor e o desenvolvimento das capacidades leitoras dos alunos, através do ensino explícito de estratégias de compreensão de textos. O estudo empírico sustentou-se em autores como Alarcão (2002), Colomer & Camps (2008), García (1992), Lomas (2003), Pawlas, & Oliva, (2007), Sim-Sim (2007), entre outros. No estudo optou-se por uma metodologia mista, qualitativa e quantitativa. O caso em estudo, sobre o PNEP e a sua influência na formação docente, concretizou-se em contexto escolar e circunscreveu-se ao cenário profissional da investigadora. Durante o ano letivo 2009/10 sete professoras/formandas, sob a supervisão da investigadora/formadora, desenvolveram a sua prática pedagógica, num Agrupamento de Escolas na área de Gondomar. Assim, foram analisadas as reflexões contidas nos portefólios formativos das docentes. Em extensão, foi aplicado um inquérito por questionário, a cento e vinte e um professores dos Núcleos Regionais do Porto e Minho. Segundo os resultados obtidos, concluiu-se que a frequência do PNEP contribuiu para formar professores reflexivos que, ao mudarem práticas, potenciaram o desenvolvimento da competência de leitura dos alunos do 1.º CEB, o que se refletiu na melhoria dos resultados escolares, em todas as áreas do saber.