A comprehensive study of herbal blends from portuguese smart-shops: isolation, analysis and toxicological impact

Dissertação de mestrado Erasmus Mundus para obtenção de grau de mestre em Técnicas Laboratoriais ForensesIn the last few years, several samples were found to contain different combinations of multiple synthetic cannabinoids, fact which may end up in a serious health threat since these substances are not fully studied and the toxic as well as the pharmacological effects are not known. The isolation and quantification of psychoactive substances from the herbal blends, known as ‘Spice’, was performed by HPLC-UV and the identification by LC-MS/MS. The HPLC-UV method was validated for the quantification and has proven to be fit for purpose. The sample 1 has shown the coexistence of JWH-122 (29,07mg/g) and JWH-210 (175,9mg/g) while the sample 2 was found to contain one single synthetic cannabinoid, the JWH-018 (149,7mg/g). The sample 3 presented the same multiple psychoactive substances as sample 1, JWH-122 (7,3mg/g) and JWH-210 (129,9mg/g). Since these drugs are mainly smoked, a smoking machine was created* in order to characterize the chemical composition of the pyrolysis. The GC-MS analysis performed to the pyrolysis solutions has shown no degradation of the JWH-018, drug used for the assay. The toxicity impact of the characterized substances was assessed on human neuroblastoma SH-SY5Y cells, applying a combination of multiple synthetic cannabinoids (JWH-122 and JWH-210) and single one (JWH-018). The preliminary results indicate that these psychoactive substances are not toxic to the cells in the concentrations ranging from 0μM to 50μM. *Designed by Dr. Alexandre Quintas and is under a patenting proces

The impact of career attitudes on developmental relationships

Organizations are currently facing a new work paradigm through global competition, restructuring, fast technological changes and restricted resources in which the constant change and adaptation has become a very important point (Hall, 1996; Dominguez & Hager, 2013). Therefore, not only the conceptualization of careers was transformed but also the mentoring literature has matured (Sullivan, 1999; Higgins & Kram, 2001). The new organizational environment is steadily gaining more and more importance, being the protean and the boundaryless considered as two popular career attitudes (Briscoe, Hall, & DeMuth, 2006). This cross-sectional study approaches how career attitudes impact developmental relationships and mentoring functions - career support, psychosocial support and role modeling. The data for this study was obtained through an online survey with a sample of 207 working professionals from different organizations and sectors. Overall, results did not verify the hypothesis proposed. However, a significant result revealed that individuals who have a boundaryless career attitude perceive to receive less career support. Thus, we discuss the implications of the findings for both theory and practice.Atualmente, as organizações encontram-se a lidar com um novo paradigma em contexto de trabalho através da concorrência global, restruturação, rápidas e constantes mudanças tecnológicas, e recursos limitados, pelo que a necessidade de constante mudança e adaptação se tem tornado um fator cada vez mais importante para as organizações (Hall, 1996; Dominguez & Hager, 2013). Desta forma, não só a conceptualização das carreiras se tem transformado, como também a literatura sobre o mentoring se tem consolidado (Sullivan, 1999; Higgins & Kram, 2001). O novo clima organizacional tem ganho constante importância pelo que as carreiras proteana e sem fronteiras têm vindo a ser consideradas como as atitudes de carreira mais estudadas (Briscoe, Hall, & DeMuth, 2006). Este estudo transversal aborda a forma como as atitudes de carreira podem ter impacto nas relações de desenvolvimento e nas funções do mentoring – suporte de carreira, suporte psicossocial e modelação de função. Os dados para o estudo foram obtidos através de um questionário online com uma amostra de 207 profissionais de diferentes organizações e sectores de atividade. Em geral, os resultados não verificaram as hipóteses propostas, no entanto, um resultado significativo revelou que indivíduos que têm uma atitude de carreira sem fronteiras consideram receber menos suporte de carreira. São discutidas as implicações dos resultados do estudo para a teoria e para a prática

Hyperhomocysteinemia: how does it affect the development of cardiovascular disease?

Homocysteine is an amino acid with an SH group, metabolised by the remethylation and transsulfuration pathways. Several genetic and environmental factors (like deficient nutrition status, systemic disease or consumption of certain drugs), can lead to changes in the levels of plasma homocysteine. Nowadays, hyperhomocysteinemia is considered an important and independent risk factor for atherosclerosis and cardiovascular disease. Several pathological mechanisms have been proposed for the effect of hyperhomocysteinemia in the development of cardiovascular disease. Among them are DNA methylation, decreased protein S-nitrosylation, production of reactive oxidative species and decrease in nitric oxide formation. Main strategies being tested for the treatment of this condition involve supplementation of folic acid, vitamins B6, B12 or riboflavin. From these, increased plasma folic acid levels by folate-rich diet or pharmacological supplementation seems to be the most effective.info:eu-repo/semantics/publishedVersio

Experimental DNA - or RNA-Directed therapies for Trinucleotide Repeat Disease

Some repeats of three or more nucleotides in tandem, which are present in a gene or in its vicinity, tend to increase in number and for this reason are called dynamic mutations. These triplet repeats are unstable and can expand from one generation to the next. According to the expansion size, an unaffected individual can carry a pre-mutation that will expand through generations leading to the development of triplet repeat expansion diseases. The increase in the number of repeats over time leads to earlier development and increased severity of symptoms in affected individuals in successive generations. Although there is still no treatment for this type of disease, several strategies are under investigation. Here, we describe treatment approaches for triplet repeat expansion diseases that have been developed over recent years, using DNA or RNA molecules as targets. Some of these strategies have the potential for future use in gene therapy for trinucleotide repeat disorders.info:eu-repo/semantics/publishedVersio

ABCDEF bundle as a tool for the prevention of Post-Intensive Care Syndrome a systematic literature review

Background: PICS (Post-Intensive Care Syndrome) refers to cognitive, physical, and psychological deficits that can develop in patients and their families during an intensive care unit (ICU) stay and persist after that. The ABCDEF bundle is a guide for the approach of critically ill patients that, by addressing pain, ventilation, analgesia and sedation, delirium, mobility, and patient’s family engagement can have a role in preventing PICS. The increasing survival rate in intensive care patients, due to the development in medicine through the years, has led to an increasing concern to discover ways to mitigate PICS triggers. Thus, we consider it to be relevant to understand the scientific base that studies the relationship between the use of the ABCDEF bundle in ICUs and the frequency of PICS in its survivors. Methods: This article follows the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-analyses) methodology. Using the PICO (participant, intervention, comparison, outcome) strategy for the literature search, articles were selected from different databases: PubMed/MEDLINE, Cochrane Library, and Science Direct. Results: Six cohort studies meeting the established inclusion criteria were included. Some studies compare different levels of adherence and total/partial performance of the bundle. Others compare the use of the bundle with the usual management of critically ill patients in a given ICU (Intensive Care Unit). The outcomes of interest for this review relate to the development of deficits in the domains involving PICS. Overall, the use of the bundle resulted in a decrease in the need for mechanical ventilation, sedation, the prevalence of delirium, and physical restriction. Conclusion: The implementation of the ABCDEF bundle seems to result in a decrease in the development of PICS triggers. Thus, since these triggers are thought to precede PICS, we could infer from our findings that the use of the bundle on critical care has a role in the prevention of PICS. It would be important to define criteria for PICS assessment to better understand the direct effect of the ABCDEF bundle on the prevention of this syndrome. Besides, it needs to be conducted more studies using the more recent bundle, including the “F” element.Introdução: A SPICI (Síndrome Pós-Internamento em Cuidados Intensivos) refere-se a défices cognitivos, físicos e psicológicos que se podem vir a desenvolver em doentes e/ou na sua família durante o internamento numa unidade de cuidados intensivos, e que persistem após esse período de internamento. Não foram estabelecidos critérios de diagnóstico para esta síndrome e não existe uma ferramenta validada para identificar os doentes críticos em risco de desenvolver SPICI. O bundle ABCDEF é uma estratégia utilizada em cuidados intensivos que segue as recomendações das guidelines PADIS (Pain, Agitation/Sedation, Delirium, Immobility, and Sleep Disruption in Adult Patients in the ICU) e que, ao incluir a abordagem da dor, ventilação, analgesia e sedação, delirium, mobilidade e envolvimento familiar do doente, pode ter um papel na prevenção da SPICI. O aumento da taxa de sobrevivência dos doentes em cuidados intensivos, devido ao desenvolvimento da medicina ao longo dos anos, tem levado a uma preocupação crescente em descobrir formas de atenuar os fatores desencadeantes da SPICI. Assim, consideramos relevante conhecer a base científica que estuda a relação entre o uso do bundle ABCDEF em doentes internados em unidades de cuidados intensivos e o desenvolvimento da SPICI nos que sobrevivem a esse internamento. Métodos: Este artigo segue a metodologia PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-analyses). Foi utilizada a estratégia PICO (população; intervenção; comparação; resultados) para a formulação da questão de investigação: “Como é que o uso do bundle ABCDEF em doentes internados em unidades de cuidados intensivos pode prevenir o desenvolvimento da SPICI?”. Para a pesquisa bibliográfica, foram selecionados artigos de diferentes bases de dados: PubMed/MEDLINE, Cochrane Library e Science Direct. Resultados: Foram incluídos seis estudos de coorte que cumpriam com os critérios de inclusão estabelecidos. Alguns estudos comparam diferentes níveis de adesão e performance total ou parcial do bundle ABCDE(F). Outros comparam a utilização do bundle com a prática comum de abordagem de doentes críticos em determinadas unidades de cuidados intensivos. Os resultados de interesse para esta revisão estão relacionados com o desenvolvimento de défices nos domínios que envolvem a SPICI. No geral, a utilização do bundle resultou numa diminuição da necessidade de ventilação mecânica, sedação, prevalência de delirium e restrição física. Conclusão: A implementação do bundle ABCDE(F) parece resultar numa diminuição do desenvolvimento de fatores desencadeantes da SPICI. Assim, uma vez que se pensa que estes fatores precedem a SPICI, podemos inferir dos nossos resultados que a utilização do bundle em cuidados intensivos tem um papel na prevenção da SPICI. No futuro, será importante definir critérios de diagnóstico da SPICI para compreender melhor o efeito direto do bundle ABCDEF na prevenção desta síndrome. Para além disso, é necessário realizar mais estudos utilizando o bundle na sua versão mais recente, com a inclusão do elemento "F"

Experimental DNA - or RNA-Directed therapies for Trinucleotide Repeat Disease

Some repeats of three or more nucleotides in tandem, which are present in a gene or in its vicinity, tend to increase in number and for this reason are called dynamic mutations. These triplet repeats are unstable and can expand from one generation to the next. According to the expansion size, an unaffected individual can carry a pre-mutation that will expand through generations leading to the development of triplet repeat expansion diseases. The increase in the number of repeats over time leads to earlier development and increased severity of symptoms in affected individuals in successive generations. Although there is still no treatment for this type of disease, several strategies are under investigation. Here, we describe treatment approaches for triplet repeat expansion diseases that have been developed over recent years, using DNA or RNA molecules as targets. Some of these strategies have the potential for future use in gene therapy for trinucleotide repeat disorders.info:eu-repo/semantics/publishedVersio

Hyperhomocysteinemia: how does it affect the development of cardiovascular disease?

Homocysteine is an amino acid with an SH group, metabolised by the remethylation and transsulfuration pathways. Several genetic and environmental factors (like deficient nutrition status, systemic disease or consumption of certain drugs), can lead to changes in the levels of plasma homocysteine. Nowadays, hyperhomocysteinemia is considered an important and independent risk factor for atherosclerosis and cardiovascular disease. Several pathological mechanisms have been proposed for the effect of hyperhomocysteinemia in the development of cardiovascular disease. Among them are DNA methylation, decreased protein S-nitrosylation, production of reactive oxidative species and decrease in nitric oxide formation. Main strategies being tested for the treatment of this condition involve supplementation of folic acid, vitamins B6, B12 or riboflavin. From these, increased plasma folic acid levels by folate-rich diet or pharmacological supplementation seems to be the most effective.info:eu-repo/semantics/publishedVersio

Macrophages in M1/M2 states and exposed to amyloid-beta: a RNA-seq analysis in human cells

Tese de mestrado em Bioinformática e Biologia Computacional, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, em 2017A Doença de Alzheimer (DA) é uma doença associada à idade e é uma das doenças neurodegenerativas com maior prevalência mundial. Esta doença causa a destruição progressiva de funções cognitivas e memória, levando à eventual morte do indivíduo. Um dos principais achados patológicos, para esta doença, são as placas extracelulares constituídas por beta amilóide (Aβ) que são encontradas frequentemente no cérbero de pacientes com DA. Enquanto que os mecanismos envolvidos na progressão desta doença permanecem desconhecidos, algumas hipóteses foram propostas envolvendo estas placas amilóides. Uma dessas hipótese é conhecida como a hipótese cascata de amiloide, e esta propõe que, através de uma cascata de eventos, a acumulação das placas de Aβ eventualmente resulta em disfunção neuronal e eventual morte celular. Microglia, os macrófagos residentes no cérbero, foram encontrados perto destas placas e já foi provado que eles têm capacidade de fagocitar estas placas. Neste trabalho foi feita uma análise de RNA-seq em dados humanos, com o objectivo de comparar os perfis genéticos e de miRNA de um grupo de controlo, de macrófagos polarizados para os fenótipos M1 e M2a e de macrófagos estimulados com Aβ. Adicionalmente, também se pretendia descobrir novas vias ou genes expressos em macrófagos presentes num ambiente com Aβ. Para tal, uma análise de expressão diferencial foi feita, tanto para os dados relativos ao mRNA como para os dados relativos ao miRNA. Uma análise do enriquecimento funcional foi feita para ajudar na compreensão dos genes diferencialmente expressos no mRNA. Por fim, uma série de testes de correlação foram feitos de modo a perceber o quão influente é a expressão de miRNA sobre o seu mRNA alvo. Através da utilização de marcadores de expressão de mRNA e miRNA, foi possível verificar a polarização dos macrófagos para M1 e M2a. Além disso, foram descobertos alguns miRNA com alta expressão nos estados de polarização, que poderiam ajudar no estudo destes estados de polarização. Relativamente ao teste de correlação, os resultados estavam enviesados para correlações positivas devido a um miRNA em particular. Uma hipótese foi criada para tentar explicar esse resultado, mas nenhuma conclusão sólida foi alcançada neste projecto.Alzheimer’s Disease (AD) is an age-related disease and it is one of the most prevalent neurodegenerative diseases in the world. This disease causes a progressive destruction of cognitive functions and memory, leading to the eventual death of the individual. One of the main pathological findings, for this disease, are the extracellular plaques of amyloid-beta (Aβ) often found in the brain of AD patients. Whilst the mechanisms involved in the progression of AD remain unknown, some hypotheses have been proposed surrounding the amyloid plaques. One such hypothesis is the Amyloid cascade hypothesis, and it purposes that, through a cascade of events, the accumulation of Aβ plaques leads to neuronal dysfunction and eventual cell death. Microglias, brain macrophages, have been found around such plaques, and it has been proven that they can phagocyte said plaques. In this present work, an analysis of human RNA-seq data was done, aiming to compare the genetic and miRNA profiles of a control group, macrophages polarized towards the M1 and M2a phenotype, and of macrophages stimulated with Aβ. Additionally, another aim was to discover of novel pathways or genes expressed in macrophages in an Aβ environment. To do so, a differential expression analysis was done for both the mRNA and miRNA data. Functional enrichment analysis was done to sort the differential expressed genes for the mRNA. Lastly, a series of correlation tests were done in order to access the amount of influence done by the expression of miRNA to their mRNA targets. It was possible to attest for the polarization of the macrophages into M1 and M2a, through the usage of mRNA and miRNA markers. Furthermore, some over-expressed miRNAs were discovered for the polarized states that could provide insight in to the study of this polarization states. For the correlation test, the results were skewed towards a positive correlation due to a single miRNA. While a hypothesis was constructed as to why it may have happened, no solid conclusion was achieved in this project thus far