In vitro comparative studies on new targets for cancer therapy, and characterization of their biomolecular effects

Dissertação de Mestrado Integrado em Medicina VeterináriaKIT is a type III receptor tyrosine kinase encoded by the KIT proto-oncogene. Mutations in KIT and dysregulation of KIT receptor have been implicated in the pathogenesis of several diseases, including human gastrointestinal stromal tumours and canine mast cell tumours, being this latter one of the most frequent cutaneous neoplasms in the dog. Despite the relatively successful use of tyrosine kinase inhibitors in the treatment of this disease, drug resistance frequently develops over time. G-quadruplexes are DNA secondary structures; potential G-quadruplex-forming sequences have been identified in the promoter region of several human oncogenes that are associated with cell growth and proliferation, including human KIT and, recently, its canine counterpart. Increasing evidence points to the potential role of G-quadruplexes in regulation of gene expression. Accordingly, the selective targeting and stabilization of G-quadruplexes by small molecule inhibitors has been studied as a novel anticancer approach. In the present dissertation, the time and dose-dependent effects of three candidate small molecule inhibitors on the expression of KIT, as well as of other proto-oncogenes comprising G-quadruplexes within their promoter regions were assessed. Two disubstituted amido-anthraquinones (compounds 1 and 2), and one bisantrene-analogue (compound 3) were studied, in two human cell lines (MCF7 and HGC27). The obtained results suggest that compound 1 was the most effective in stabilizing KIT G-quaduplex structures and inhibiting mRNA expression; furthermore, the transcriptional effect was confirmed at the protein level. The most pronounced modulation at the protein level was achieved by compound 3. Compound 2 did not cause significant changes in the expression of any of the investigated proto-oncogenes. All in all, the direct targeting of KIT proto-oncogene G-quadruplexes with compounds 1 and 3 might be a promising treatment alternative to the tyrosine kinase inhibitors, in both human and canine KIT-related tumours and, particularly, canine mast cell tumours.RESUMO - Estudos comparativos in vitro de novos alvos para terapia do cancro, e caracterização dos seus efeitos biomoleculares - O KIT é um recetor de tirosina quinase tipo III codificado pelo proto-oncogene KIT. A ocorrência de mutações no KIT e a desregulação do recetor KIT têm sido implicadas na patogénese de diversas doenças, incluindo os tumores do estroma gastrointestinal no homem e os mastocitomas caninos, sendo estes últimos uma das neoplasias cutâneas mais frequentes no cão. Os inibidores da tirosina quinase têm sido usados com relativo sucesso no tratamento desta doença, contudo com o tempo desenvolve-se, frequentemente, resistência a estes fármacos. Os G-quadruplexes são estruturas secundárias de ADN. Na região promotora de vários oncogenes humanos, que estão associados a crescimento e proliferação celular e que incluem o KIT humano e, recentemente, o seu congénere canino, têm sido identificadas potenciais sequências formadoras de G-quadruplex. Evidências crescentes apontam para o papel potencial dos G-quadruplexes na regulação da expressão génica. Nesse sentido, a atuação seletiva e estabilização de G-quadruplexes por meio de pequenas moléculas inibidoras tem sido estudada como uma nova abordagem anti tumoral. Na presente dissertação foram avaliados os efeitos, dependentes do tempo e da concentração, de três moléculas possivelmente inibidoras na expressão do KIT, bem como de outros proto-oncogenes que contêm G-quadruplexes nas suas regiões promotoras. Assim, foram estudadas duas amido-antraquinonas dissubstituídas (compostos 1 e 2), e um análogo do bisantreno (composto 3), em duas linhas celulares humanas (MCF7 e HGC27). Os resultados obtidos sugerem que o composto 1 foi o mais eficaz na estabilização de estruturas G-quadruplex do KIT e na inibição da expressão de ARNm, tendo o efeito transcricional sido confirmado a nível proteico. A modulação mais acentuada a nível proteico foi alcançada pelo composto 3. O composto 2 não causou alterações significativas na expressão de qualquer um dos proto-oncogenes avaliados. Em conclusão, a atuação direta dos compostos 1 e 3 sobre os G-quadruplexes no proto-oncogene KIT poderá ser uma promissora alternativa de tratamento aos inibidores da tirosina quinase em tumores humanos e caninos associados ao KIT, e em mastocitomas em particular

3D Construction Printing Standing for Sustainability and Circularity: Material-Level Opportunities

Three-dimensional Cementitious materials Printing (3DCP) is a cutting-edge technology for the construction industry. Three-dimensional printed buildings have shown that a well-developed automated technology can foster valuable benefits, such as a freeform architectural design without formworks and reduced human intervention. However, scalability, commercialization and sustainability of the 3DPC technology remain critical issues. The current work presents the ecological fragility, challenges and opportunities inherent in decreasing the 3DCP environmental footprint at a material level (cementitious materials and aggregates). The very demanding performance of printable mixtures, namely in a fresh state, requires high dosages of cement and supplementary cementitious materials (SCM). Besides the heavy carbon footprint of cement production, the standard SCM availability might be an issue, especially in the longer term. One exciting option to decrease the embodied CO2 of 3DCP is, for example, to incorporate alternative and locally available SCM as partial cement replacements. Those alternative SCM can be wastes or by-products from industries or agriculture, with no added value. Moreover, the partial replacement of natural aggregate can also bring advantages for natural resource preservation. This work has highlighted the enormous potential of 3DCP to contribute to reducing the dependence on Portland cement and to manage the current colossal wastes and by-products with no added value, shifting to a Circular Economy. Though LCA analysis, mixture design revealed a critical parameter in the environmental impact of 3DCP elements or buildings. Even though cement significantly affects the LCA of 3DCP, it is crucial to achieving adequate fresh properties and rheology. From the literature survey, mixtures formulated with alternative SCM (wastes or by-products) are still restricted to rice husk ash, Municipal Solid Waste ashes and recycled powder from construction and demolition wastes. Natural aggregate replacement research has been focused on recycled fine sand, mine tailing, copper tailing, iron tailing, ornamental stone waste, recycled glass, crumb rubber, rubber powder and granules, recycled PET bottles and steel slag. However, flowability loss and mechanical strength decrease are still critical. Research efforts are needed to find low-carbon cement replacements and mix-design optimization, leading to a more sustainable and circular 3DCP while ensuring the final product performance

Urinary tract infections on the Portuguese general practitioner sentinel network - preliminary data 2016

Urinary tract infections (UTIs) are inflammatory processes of infectious cause, which can reach the upper and/or lower urinary tract. UTIs, after respiratory infections, are the most frequent in the community, with a higher incidence in females. They can be classified according to anatomical location: in cystitis (low UTI) where the presence of bacteria is limited to the bladder; and pyelonephritis (high UTI) defined as that affecting the pelvis and renal parenchyma. Escherichia coli is responsible for 75 to 90% of the cases. The high rates of resistance of E. coli to quinolones and co-trimoxazol described in Portugal are likely to be a consequence of the prescription abuse of these antibiotics over the last few decades and may increase the risk of antibiotic failure. When the antibiotic prescribing decision is based on the knowledge of the main microbial agents involved and its sensitivity profile is known, allowing guidelines to be adjusted to the current reality, empirical antibiotic therapy is associated with eradication rates equal to or greater than 90%.N/

O efeito da proibição de serviços cruzados pelas auditoras no planeamento fiscal das empresas europeias

Mestrado em Contabilidade, Fiscalidade e Finanças EmpresariaisEste estudo tem como finalidade perceber qual o efeito da proibição de serviços cruzados prestados pelas auditoras, imposta pelo Regulamento (UE) n.º 537/2014, no planeamento fiscal das empresas abrangidas pelo mesmo. Dado que a taxa efetiva de imposto é uma medida usada para avaliar a eficácia do planeamento fiscal, foi analisada a sua evolução tanta nas empresas cotadas europeias, proxies das empresas sujeitas ao regulamento, como nas empresas não cotadas europeias, não sujeitas ao regulamento, a fim de deduzir se houve ou não uma minimização da carga fiscal. O modelo econométrico utilizado foi o difference-in-differences, que permite, através da interação entre uma variável de tempo e de grupo, averiguar se a diferença nesta taxa entre os dois grupos se altera aquando uma comparação entre o período pré-regulamento e o período pós-regulamento, e concluir sobre o impacto que a proibição destes serviços teve na criação de valor das empresas. Os resultados sugerem que o regulamento trouxe consigo um agravamento fiscal que não passou por um aumento no imposto pago, mas sim pela restrição de beneficiarem de uma maior redução da TEI. Assim, é legitimo afirmar que a imposição deste regulamento afetou a criação de valor destas empresas, uma vez que se observou que a TEI poderia ter sido mais baixa do que a que foi na realidade. O presente estudo contribui para esclarecer que existem vantagens para uma empresa em receber em simultâneos diversos serviços da mesma entidade e que a sua proibição provoca um impacto negativo na criação de valor.This study aims at understanding which effect is linked to the prohibition on cross-services by audit firms, imposed by the Regulation (EU) number 537/2014, in the tax planning of companies that are subjected to this regulation. Since the effective tax rate is a widely used measure to evaluate tax planning effectiveness, its evolution was analysed both in European listed companies, proxies of companies subjected to the Regulation, and in European unlisted companies, not subjected to the Regulation, in order to deduce whether or not the tax burden was minimized. The econometric model used was the difference-in-differences, which allows, through the interaction between a time and group variable, to determine if the difference in this rate between the two groups changes when comparing the pre-regulation period and the post-regulation period, and conclude on the impact that the prohibition of these services had on firm´s value creation. The results suggest that the regulation is associated with a tax worsening related to the restriction in benefiting of a greater reduction of TEI and not connected to an increase of taxes. Thus, it is possible to state that the imposition of this regulation affected the value creation of these companies since it was observed that the TEI could have been lower compared to the actual result. The present study contributes to clarify that there are advantages for a company to simultaneously receive several services from the same entity and that its prohibition has a negative impact on value creation.info:eu-repo/semantics/publishedVersio

Infeções do Trato Urinário na Rede Médicos Sentinela – dados preliminares de 2016

INTRODUÇÃO:As infeções do trato urinário (ITU) são processos inflamatórios de causa infeciosa, que podem atingir as vias urinárias superiores e/ou inferiores. As ITU, depois das infeções respiratórias, são as mais frequentes na comunidade, apresentando maior incidência no sexo feminino. As bactérias são os principais agentes responsáveis, dos quais a Escherichia coli é responsável por 75 a 90% dos casos. Quando a decisão de prescrição do antibiótico se fundamenta no conhecimento dos principais agentes microbianos envolvidos e se conhece o seu perfil de sensibilidade a antibioterapia empírica associa-se a taxas de erradicação iguais ou superiores a 90%. Tal permite também realizar guidelines ajustadas à realidade atual. O objetivo deste estudo foi conhecer o perfil de orientação e prescrição dos casos de ITU notificados pelos médicos da Rede Médicos Sentinela. METODOLOGIA: Estudo descritivo realizado na população sob observação da Rede Médicos Sentinela durante o ano de 2016 (entre 1 de janeiro e 20 de setembro). Foi pedido aos médicos desta Rede que notificassem todos os casos de ITU diagnosticados na sua lista de utentes em 2016. Para cada caso um conjunto de variáveis demográficas, clínicas e microbiológicas foi colhido através de um questionários estruturado enviado em papel ou através de uma plataforma online. Foi calculada a proporção de casos por sexo, grupo etário, tipo de infecção, antibiótico prescrito e microrganismo identificado. RESULTADOS: Foram identificados 360 ITU. 87 % dos casos era do sexo feminino. A idade mediana foi de 62 anos, sendo que 27,8 % dos casos ocorreu no grupo etário com 75 ou mais anos. A maioria das ITU (83,6%) foram classificadas como cistites, 13,6% bacteriúrias, 2,5% pielonefrites ligeiras e 0,3% (1 caso) como pielonefrite complicada. 8 casos bacteriúria ou cistite ocorreram em grávidas. Foi efectuada urocultura em 46% (167 casos), sendo que 99 % destas foram positivas. Os microrganismos mais frequentes foram E. Coli (81%) e Proteus (5%). Foram prescritos antibióticos a 98,6%, sendo a fosfomicina (47,6%), a amoxicilina e ácido clavulânico (15,8%) e a nitrofurantoína (10,7%) os mais prescritos. O tempo médio de tratamento foi de 2 dias (fosfomicina) e de 8 dias (amoxicilina e nitrofurantoína). DISCUSSÃO: Embora os resultados obtidos sejam ainda preliminares, apontam que em Portugal, nas ITU da comunidade, a maioria dos casos são diagnosticadas como cistite, o microrganismo mais frequentemente identificado é E. Coli e o antibiótico mais prescrito é a fosfomicina.N/

Peripheral Biomarkers in Multiple Sclerosis Patients Treated with Interferon-Beta

Multiple sclerosis is a relapsing and eventually progressive disorder of the central nervous system that continues to challenge researchers who try to understand the pathogenesis of the disease and prevent its progression. Interferon-beta is the most widely prescribed treatment for MS. Peripheral blood seems to mirror the immunological disturbances that underlie MS, which could represent the migration patterns between periphery and other tissues according to the clinical phase of the disease. Based on this assumption, several studies point to significant alterations in peripheral blood homeostasis of different subpopulations of T cells, like γδ T cells or Th1, Th2 and Th17 functional subsets; of B cells subpopulations; and of innate cells like monocytes and dendritic cells. The main goal of this chapter is to make an in-depth review of the major findings described in the literature that correlate specific alterations on different leukocytes subpopulations with disease status, and which therefore have the potential to constitute a peripheral biomarker of disease progression

Interferon-beta treated-multiple sclerosis patients exhibit a decreased ratio between immature/transitional B cell subset and plasmablasts

Our aim was to quantify circulating B cell subsets; immature/transitional, naïve, CD27- and CD27+ memory cells and plasmablasts, in relapsing-remitting multiple sclerosis patients treated with IFN-β. The most relevant findings were a significant increase of plasmablasts and a decrease of immature/transitional B cells, resulting in a decreased ratio between those cells in relapse RRMS, together with an increase of CD27- and CD27+IgM+ memory B cell subsets in both phases of the disease. These alterations point to an active B cell response, particularly in relapse, and the above referred ratio could constitute a good biomarker of relapse in patients that underwent IFN-β treatment.info:eu-repo/semantics/publishedVersio

Molecular survey of vector-borne diseases in two groups of domestic dogs from Lisbon, Portugal

Research Areas: Parasitology ; Tropical MedicineABSTRACT - Background: Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. Methods: One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. Results: Infections were detected by PCR in 48 (33.80%) dogs. Single infections were found in 35 dogs (24.64%), and co-infections were found in 13 (9.15%) dogs. Twenty-nine (20.42%) dogs were positive for Hepatozoon spp., 15 (10.56%) for Mycoplasma spp., 11 (7.75%) for Anaplasma spp./Ehrlichia spp., and six (4.21%) for Babesia spp. DNA sequencing was used to identify Babesia vogeli (2.81%), Babesia canis (1.40%), Hepatozoon canis (20.42%), Mycoplasma haematoparvum (2.11%), Mycoplasma haemocanis (8.45%), Anaplasma platys (7.04%), and Ehrlichia canis (0.70%). Conclusions: This is the frst molecular identifcation of B. canis and M. haematoparvum in dogs from southern Portugal. This study highlights the importance of molecular methods to identify CVBD pathogens in endemic areas and helps to guide the clinical approach of veterinarians in practice.info:eu-repo/semantics/publishedVersio

Evolução da taxa de incidência de depressão e do desemprego em Portugal entre 1995–2013: dados da Rede Médicos Sentinela

OBJECTIVO: Quantificar, para ambos os sexos, a correlação entre a taxa de incidência de depressão e a taxa de desemprego, em Portugal, entre 1995 e 2013. MÉTODOS: Foi desenvolvido um estudo ecológico no qual se correlacionou a evolução das taxas de incidência de depressão estimadas pela Rede Médicos Sentinela e as taxas de desemprego anuais disponibilizadas pelo Instituto Nacional de Estatística em publicações oficiais. RESULTADOS: Observou-se uma correlação positiva entre taxa de incidência de depressão e taxa de desemprego em Portugal, sendo esta significativa apenas para o sexo masculino (R2 = 0,83; p = 0,04). Estimou-se, para este sexo, um aumento de 37 novos casos de depressão por 100.000 habitantes a cada 1% de aumento da taxa de desemprego entre 1995 e 2013. CONCLUSÕES: Embora o desenho do estudo não permita o estabelecimento de uma relação causal entre desemprego e depressão, os resultados obtidos sugerem que a evolução do desemprego em Portugal poderá ter tido um impacto não desprezável no nível de saúde mental dos portugueses, em especial no sexo masculino.OBJECTIVE: Quantify, for both genders, the correlation between the depression incidence rate and the unemployment rate in Portugal between 1995 and 2013. METHODS: An ecological study was developed to correlate the evolution of the depression incidence rates estimated by the General Practitioner Sentinel Network and the annual unemployment rates provided by the National Statistical Institute in official publications. RESULTS: There was a positive correlation between the depression incidence rate and the unemployment rate in Portugal, which was significant only for males (R2 = 0.83, p = 0.04). For this gender, an increase of 37 new cases of depression per 100,000 inhabitants was estimated for each 1% increase in the unemployment rate between 1995 and 2013. CONCLUSIONS: Although the study design does not allow the establishment of a causal association between unemployment and depression, the results suggest that the evolution of unemployment in Portugal may have had a significant impact on the level of mental health of the Portuguese, especially among men

Long-term intestinal failure and home parenteral nutrition: a single center experience

Communication abstract: Proceedings of the 5th International Congress of CiiEM - Reducing inequalities in Health and Society, held at Egas Moniz’ University Campus in Monte de Caparica, Almada, from June 16th to 18th, 2021.This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Intestinal failure is the reduction in gut function below the minimum necessary for the absorption of macronutrients and/or water electrolytes. The based treatment for type II and III intestinal failure patients is home parenteral nutrition (HPN) and hydration (HPH). This is a case-series study of HPN/HPH patients of the Hospital Garcia de Orta, Portugal, where thirteen patients present different underlying disorders and various IVS needs of nutrition and/or hydration. Most presented type III failure and most of them survived a long period under HPN and/or HNH.info:eu-repo/semantics/publishedVersio