VCAM1 modulation on endothelial cells – implications for vasculopathy in sickle cell anemia

Sickle cell anemia (SCA) is a highly heterogeneous and multifactorial-like monogenic disease that arises from homozygosity for the c.20A>T mutation in the HBB gene. Vascular disease is systemic in SCA, with profound effects in organs like the brain, where stroke is the most severe end of the cerebral vasculopathy (CVA) spectrum. Endothelial dysfunction plays a major role in vasculopathy with several adhesion molecules, such as VCAM1, being produced by the endothelium altered as a response to inflammatory cytokine (e.g., TNF-α) signalling. In previous association studies, we found positive associations between the presence of four specific VCAM1 gene promoter haplotypes and i) high blood flow velocities in the median cerebral artery, and ii) a chronic hemolysis biochemical marker. In this study, we aimed to assess the functional role of those VCAM1 promoter haplotypes in endothelial cell response following endothelial activation through TNF-α stimulation. After molecular cloning of 3 haplotype constructs, using a pGL4 promoterless vector, haplotype sequence was confirmed, by Sanger sequencing, prior to transfection. We used EAhy926, HUVEC and HBEC as different endothelial cell models, and performed transfection experiments for each construct, with and without TNF-α stimulation. Differences in promoter activity were assessed by luciferase reporter assay. All haplotypes showed differences in promoter activity, after TNF-α stimulation, in all cell models. Haplotype 1 showed decreased promoter activity, while haplotypes 7 and 8 showed increased activity after TNF-α stimulation, in all cell models. These results are consistent with lower VCAM1 expression due to haplotype 1, and therefore a protective effect. Conversely, a higher expression due to haplotypes 7 and 8, suggests an increased vasculopathy risk, in a pro-inflammatory milieu. The association between specific haplotypes and endothelial cell response further enhances the modifier effect of VCAM1 on endothelial dysfunction and its impact in SCA pathophysiology, as well as its potential role as a biomarker of SCA vasculopathy risk, severity and prognosis.This work was partially supported by INSA_202DGH720 project.info:eu-repo/semantics/publishedVersio

Cerebral vasculopathy in children with sickle cell disease A study of genetic modulators of the disease

Sickle cell disease (SCD) is a genetic disorder with recessive transmission, caused by the mutation HBB:c.20A>T. It originates hemoglobin S that forms polymers inside the erythrocyte, upon deoxygenation, deforming it and ultimately leading to premature hemolysis. The disease presents with high heterogeneity of clinical manifestations, the most devastating of which, ischemic stroke, occurs in 11% of patients until 20 years of age. In this study, we tried to identify genetic modifiers of risk and episodes of stroke by studying 66 children with SCD, grouped according to the degree of cerebral vasculopathy (Stroke, Risk and Control). Association studies were performed between the three phenotypic groups and hematological and biochemical parameters of patients, as well as with 23 polymorphic regions in genes related to vascular cell adhesion (VCAM-1, THBS-1 and CD36), vascular tonus (NOS3 and ET-1) and inflammation (TNF-α and HMOX-1). Relevant data was collected from patient’s medical records. Known genetic modulators of SCD (beta-globin cluster haplotype and HBA and BCL11A genotypes) and putative genetic modifiers of cerebral vasculopathy were characterized. Differences in their distribution among groups were assessed. VCAM-1 rs1409419 allele C and NOS3 rs207044 allele C were associated to stroke events, while VCAM-1 rs1409419 allele T was found to be protective. Alleles 4a and 4b of NOS3 27 bp VNTR appeared to be respectively associated to stroke risk and protection. HMOX-1 longer STRs seemed to predispose to stroke. Higher hemoglobin F levels were found in Control group, as a result of Senegal haplotype or of BCL11A rs11886868 allele T, and higher lactate dehydrogenase levels, marker of hemolysis, were found in Risk group. Molecular mechanisms underlying the modifier functions of the relevant genetic variants are discussed

Differential Endothelial VCAM1 Expression and Implications for Sickle Cell Anemia Vasculopathy

Comunicação oral por conviteBackground: Vascular disease is systemic in sickle cell anemia (SCA), with profound effects in organs like the brain,where stroke is the most severe end of the cerebral vasculopathy spectrum. Endothelial dysfunction is an important pathobiological mechanism in SCA systemic vasculopathy, with upregulation of adhesion molecules (e.g., VCAM-1), lower nitric oxide bioavailability, and increased oxidative stress. In previous association studies, we found positive associations between the presence of three specific VCAM1 gene promoter haplotypes and i) high blood flow velocities in the median cerebral artery, and ii) a chronic hemolysis biochemical marker. Aims: The aims of our work were: a) to investigate the functional role of those VCAM1 promoter haplotypes in endothelial cell response following endothelial activation through TNF-α stimulation; b) to assess the modulation role of proinflammatory and/or pro-oxidative stimuli on endothelial VCAM1 expression; and, finally, to evaluate how hydroxyurea (HU) treatment would affect that expression. Methods: After molecular cloning of three VCAM1 promoter haplotype constructs, using pGL4 promoterless vectors, haplotype sequence was confirmed, by Sanger sequencing, prior to transfection. Transfection experiments for each construct were performed, with or without TNF-α stimulation, using EAhy926, and HBEC as macrovascular and microvascular endothelial cell models, respectively. Differences in promoter activity were assessed by luciferase reporter assay. RNA was extracted from non-transfected EAhy926 and HBEC cell cultures stimulated or not with TNF-a and/or hemin, and with or without HU treatment. RT-qPCR was performed to analyze VCAM1 expression. HMOX1 and NOS3 were also analyzed for comparison purposes. Results: Our results showed that two VCAM1 promoter haplotypes, previously associated with pediatric cerebral vasculopathy and hemolysis in SCA, increased promoter activity in transfected and TNF-α-stimulated EA.hy926 and HBEC cells, consistent with a higher VCAM1 expression in macro and microvascular settings. In non-transfected cells, we also observed TNF-a-induced VCAM1 overexpression as well as heme-induced overexpression of HMOX1 in both cell models. Heme did not affect VCAM1 nor NOS3 expression and the latter was also not affected by TNF-a stimulus. Hydroxyurea treatment lowered TNF-a-induced VCAM1 and NOS3 expression but did not affect heme-induced HMOX1 expression. Summary/Conclusion: These data further indicate that VCAM1 haplotypes we previously associated with pediatric cerebral vasculopathy and hemolysis in SCA, induce higher VCAM1 expression potentially affecting both cerebral and systemic vasculopathy risk. The differential endothelial expression of VCAM1, NOS3, and HMOX1 after proinflammatory and/or pro-oxidative stimuli also reinforces their genetic modulation role in SCA systemic vasculopathy.N/

Caracterização de suportes em papel vegetal

O presente Relatório de Estágio é fruto de um estudo sobre suportes em papel vegetal. O estudo incide sobre vinte Desenhos de Arquitectura de edifícios da Cidade de Lisboa e cinco desenhos de decalque para cerâmicas, caracterizando os vinte e cinco documentos através da identificação da sua composição e metodologia de produção. A caracterização dos suportes em papel vegetal estudados teve como referência e fundamento os resultados obtidos por diversos exames e análises, nomeadamente exames estratigráficos, testes microquímicos de detecção de amido e lenhina, Espectroscopia de Infravermelho com Transformada de Fourier (FTIR) e Cromatografia Gasosa com Espectrómetro de Massa e Hidrólise termicamente assistida (THM-GC/MS), que permitiram a identificação dos papéis vegetais impregnados. Pode-se com isto verificar, neste grupo de documentos em estudo, uma predominância do uso dos papéis vegetais impregnados em relação aos obtidos pelas restantes metodologias de produção. Englobadas no âmbito de um trabalho em Arquivo, são ainda apresentadas outras intervenções de Conservação e Restauro, com especial incidência na pesquisa de procedimentos de remoção de fitas-cola e de reforços parciais de áreas fragilizadas. Estas acções foram efectuadas em dez Desenhos de Arquitectura do Arquivo Municipal de Lisboa e também noutros documentos gráficos como tela imperial e suportes em pasta de madeira. PalavrasInstituto Politécnico de Toma

A Informação: Desafio Ético na Era da Globalização Contemporânea

info:eu-repo/semantics/publishedVersio

O problema da verdade em Kant

This article aims to elucidate how the problem of truth arises in Immanuel Kant alongside the question of the criterion of truth, as well as pointing out its predominance. Based on the answer to this question we can avoid incurring in circles of which would become impossible to get out, regarding the author. The problem looms on how to know what is the criterion of truth? The reflections lead us to Kritik der reinen Vernunft and Prolegomena zu einer jeden kün ftigen Metaphysik, die als Wissenschaft wird auftreten können, but essentially to Logik: ein Handbuch zu Vorlesungen.O presente artigo visa elucidar como o problema da verdade surge em Immanuel Kant a par da pergunta pelo critério de verdade, bem como assinalar a predominância deste. É com base na resposta a esta questão que, quanto ao autor, poderemos evitar incorrer em círculos dos quais se tornaria impossível sair. O problema assoma em como saber qual o critério de verdade? As reflexões remetem-nos para Kritik der reinen Vernunft e Prolegomena zu einer jeden kün ftigen Metaphysik, die als Wissenschaft wird auftreten können, mas essencialmente, para Logik: ein Handbuch zu Vorlesungen

Experiência profissionalizante na vertente de farmácia comunitária e investigação

Esta dissertação encontra-se dividida em dois capítulos, sendo o primeiro respeitante à componente de investigação e o segundo respeitante à experiência profissionalizante em farmácia comunitária. O capítulo I concretiza uma abordagem psicofarmacológica dos alucinogénios clássicos, com foco nos seus potenciais efeitos terapêuticos. Um alucinogénio carateriza-se por, após administração de uma dose única, produzir alterações no pensamento, no estado de espírito e na perceção, com poucas afetações da memória, produzir narcose (sonolência, com perda de sensibilidade geral ou local), ou estimulação excessiva. Os alucinogénios clássicos definem-se por exercerem agonismo dos recetores de serotonina (5-HT), particularmente dos recetores do tipo 5-HT(2A). Atualmente, os alucinogénios clássicos estão cada vez mais a ser estudados no âmbito da saúde mental. Embora ainda não existam alucinogénios clássicos aprovados para o tratamento de doenças psiquiátricas, perspetiva-se uma mudança de paradigma. De um total de 59 inquiridos, 32% dos profissionais de saúde argumentou que estas substâncias podem vir a desempenhar um papel preponderante em psiquiatria, enquanto 44% dos profissionais não acreditam que esta seja uma opinião viável. Os restantes 24% não tiveram qualquer opinião sobre o assunto. O capítulo II é respeitante ao estágio feito em farmácia comunitária, durante a fase inicial da pandemia de Coronavirus disease 19. A farmácia comunitária constitui, por si, um dos pontos iniciais e mais habituais de contato entre os doentes e os sistemas de saúde. No entanto, no contexto de uma pandemia global, torna-se fulcral no apoio às populações.This dissertation is divided in two chapters, the first concerning the research component and the second concerning the professional experience in community pharmacy. Chapter I provides a psychopharmacological approach to classic hallucinogens, focusing on their potential therapeutic effects. A hallucinogen is characterized by, after administration of a single dose, producing changes in thought, mood and perception, with few changes in memory, producing narcosis (drowsiness, with loss of general or local sensitivity), or excessive stimulation. Classic hallucinogens are defined by exerting agonism on serotonin (5-HT) receptors, particularly 5-HT (2A) type receptors. Nowadays, classic hallucinogens are being studied in the context of mental health. Although there are still no classic hallucinogens approved for the treatment of psychiatric illnesses, a paradigm shift is expected. Of a total of 59 respondents, 32% of health professionals argued that these substances may play a leading role on psychiatry, while 44% do not believe this is a viable opinion. The remaining 24% had no opinion on the matter. Chapter II concerns the internship in community pharmacy, during the initial phase of Coronavirus disease 19 pandemic. Community pharmacy is one of the initial and most common points of contact between patients and the health systems. However, in the context of a global pandemic, it becomes crucial in providing support to populations

Influência do mineral precursor no uso de uma bactéria para reparação de fissuras no betão

Nesta dissertação foi feita uma avaliação da influência do mineral precursor no uso da bactéria Bacillus cohnii para reparação de fissuras no betão. Foram preparadas várias amassaduras de betão onde foram introduzidas bactérias da espécie Bacillus cohnii e combinadas com dois tipos diferentes de minerais precursores. Pretendeu-se avaliar qual o mineral precursor mais eficaz na formação de carbonato de cálcio quando combinado com esta bactéria. Através dos ensaios realizados fez-se um acompanhamento da evolução das fissuras nos betões ao longo do tempo os quais mostram que é possível colmatar em parte as fissuras e que o silicato de cálcio foi o meio precursor mais eficiente; Influência do mineral precursor no uso de uma bactéria para reparação de fissuras no betão Abstract: Influence of the mineral precursor in the use of a bacterium to heal concrete cracks In the present study the influence of the mineral precursor in the use of the bacterium Bacillus cohnii was evaluated in the healing of concrete cracks. Several concrete mixings were prepared and bacteria of the species Bacillus cohnii were introduced combined with two different types of precursor minerals. The aim was to determine the most effective precursor mineral in the formation of calcium carbonated when combined with the bacterium. The evolution of concrete cracks was monitored over time, and the performed trials showed that it is possible to partially heal cracks and that calcium silicate showed the best results, being the most effective precursor mineral

El uso del gesto y el movimiento cotidiano para facilitar una relación entre el espectador y la danza contemporánea : estudio de la obra Quita Pesar dirigida por Ana Chung

Esta investigación trata sobre el uso del gesto y el movimiento cotidiano como vínculo para generar una relación entre el espectador y la danza contemporánea. Para lograr tal propósito, se analizó la obra de danza contemporánea Quita pesar, dirigida por Ana Chung y estrenada en el 2018, la cual aborda el uso de las acciones cotidianas con el objetivo de crear reflexión sobre el cuerpo y la comunicación que el movimiento puede generar en las personas. Asimismo, se tomaron como principales referentes a la coreógrafa Pina Bausch y el colectivo de danza post moderna Judson Dance Church, ya que ambos sirvieron para indagar la causa de la aparición del gesto y el movimiento cotidiano en la danza, su historia y evolución. Como material de análisis se realizó un trabajo de campo donde se estudió cada función realizada, la opinión de los espectadores y se llevaron a cabo entrevistas a los intérpretes con el objetivo de demostrar si existe la posibilidad de generar una relación de cercanía entre el espectador y la danza contemporánea mediante el uso del gesto y el movimiento cotidiano.This research is concerned by the use of gestures and daily movements as a link to generate a relationship between the audience and contemporary dance. As an object of study, the work of contemporary dance Quita Pesar was analyzed. Directed by Ana Chung and launched in 2018, the play addresses the use of daily actions with the aim of creating a reflection on the body and communication that the movement might generate in the audience. As main references we consider the choreographer Pina Bausch and the postmodern dance collective Judson Dance Church, both were utilized to investigate the cause of the appearance of the gesture and the daily movement in dance, its history and evolution. As an analysis material, field work was carried out. We studied each performance, opinions of the spectators and interviews with the interpreters, with the aim of demonstrating whether there is a possibility to generate a close relationship between the audience and contemporary dance through the use of gestures and everyday movements.Tesi