Impact and ecological adaptation of Leptoglossus occidentalis (Hemiptera, Coreidae) in Pinus pinea

Doutoramento em Engenharia Florestal e dos Recursos Naturais - Instituto Superior de Agronomia / ULLeptoglossus occidentalis Heidemann (Heteroptera: Coreidae) is an invasive pest seriously damaging conifer seeds. Native from North America, the insect was first detected in Europe in 1999, and recorded in Portugal in 2010. Both nymphs and adults feed on seeds of several conifer species. Bug impact on seed production of Stone pine, Pinus pinea, is of major concern in the Mediterranean Basin countries because cone production and seed yield have decreased during the last years quite simultaneously to the records of L. occidentalis. Thus, the insect has been considered the most plausible cause of this decrease. However, there was still a substantial lack of knowledge about the effective impact of bugs and their ecological adaptations on Stone pine. My main goal was to add valuable and pertinent knowledge to understand the interactions between seed bugs and Stone pine. At first, a careful revision of all the literature available about Leptoglossus occidentalis was carried out, together with discussions with other European researchers working on this pest, in order to define the PhD aims. The PhD plan was then divided into three main issues. I first characterized and measured the importance of bug damage on seeds of Stone pine. In a second part, I investigated the ecological interactions between invasive bugs and Stone pine cones and seeds. Bug host preference was tested between Stone pine and the other two main native pine species growing in southern Europe (P. pinaster and P. halepensis), and cues possibly underlying such preferences were suggested. I also evaluated the impact of the bug in Stone pine seed orchards under two different management strategies. In a third part, I suggested possible invasion routes of L. occidentalis in the Iberian Peninsula, using genetic data and field recordsN/

Metaciclogénese de Leishmania sp in vitro - análise cinética e morfológica

Foi só em 1984 que, Sacks&Perkins, demonstraram in vitro e no vector o desenvolvimento de promastigotas de Leishmania tropica de um estadio não infeccioso (procíclicas) para formas infecciosas (metacíclicas). Este processo foi denominado de metaciclogénese. As formas metacíclicas são caracterizadas por possuírem um corpo celular pequeno e fino e um flagelo muito longo, e ocorrer em maior número durante a fase estacionária de crescimento em meio de cultura (Sacks&Perkins, 1984; Franke et al., 1985; Da Silva&Sacks, 1987; Sacks, 1989; Grimm et al., 1991; Saraiva et al., 2005). Diversos estudos sugerem o comprimento celular como um parâmetro eficaz na distinção das formas procíclicas das metacíclicas (Grimm et al., 1991; Saraiva et al., 2005; Späth&Beverley, 2001). Neste estudo, foram analisadas diariamente seis culturas axénicas, duas L. braziliensis (1794 e LC2452cl8), três L. infantum (IMT151, IMT260 e IMT373) e uma L. peruviana (HR78cl8) de forma a ser possível, no final, obter uma curva de crescimento para cada estirpe, isto é, a cinética e a concentração dos parasitas em função do tempo, bem como o perfil da metaciclogénese de cada uma. O crescimento foi estudado em dois meios de cultura, Schneider e Grace, ambos suplementados com 10% de FBS e pH de 5.5. Foram realizados estudos de crescimento dos parasitas em cultura e estudos morfológicos dos promastigotas sendo a variável dependente a média do comprimento dos promastigotas e tendo como variáveis independentes a estirpe, o meio de cultura e a fase de crescimento. O teste da Lise pelo Complemento foi usado como método de isolamento das formas metacíclicas em cultura (Franke et al., 1985; Puentes et al., 1988; Da Silva&Sacks, 1987, Sacks&Perkins, 1984; Sacks, 1989; Grimm et al., 1991). Os parasitas de todas as estirpes estudadas revelaram, em cultura axénica de meio Schneider, uma cinética de crescimento mais rápida, uma maior concentração de promastigotas, uma fase estacionária mais curta e um menor tempo de vida que as mesmas estirpes em meio Grace. Estes resultados sugerem que o meio de cultura usado influencia a cinética de crescimento e a longevidade de uma estirpe in vitro. Relativamente ao perfil da metaciclogénese, foi também no meio de cultura Schneider que, todas as estirpes, apresentaram populações em cultura com percentagens de formas metacíclicas mais elevadas do que no meio Grace. A maior percentagem de formas metacíclicas foi atingida, para todas as estirpes, e independentemente do meio, durante a fase de declínio da curva de crescimento apesar do máximo de concentração de promastigotas em solução corresponder ao início da fase estacionária de crescimento. Os resultados do estudo morfológico dos parasitas, mais precisamente a análise da evolução do comprimento destes ao longo da sua curva de crescimento, mostraram não existir uma diferença significativa do tamanho médio dos parasitas no meio de cultura Schneider do tamanho no meio de cultura Grace para uma mesma estirpe, levando a concluir que o meio de cultura não influencia o tamanho dos promastigotas. Contudo, no que diz respeito às fases de crescimento das culturas – fase logarítmica e fase estacionária, verificou-se, para todas as estirpes, uma diferença significativa entre o tamanho médio dos promastigotas entre as duas fases de crescimento, independentemente do meio de cultura. O tamanho médio de parasita foi, para todas as estirpes, sempre significativamente inferior na fase estacionária. Comparando os valores do comprimento médio dos promastigotas apresentados pelo teste do Complemento e os obtidos no estudo de cinética, para cada estirpe e na mesma fase de crescimento, verificou-se que foi com a aplicação do teste do Complemento que se obtiveram os valores mais baixos de tamanho médio dos parasitas, sugerindo que com a aplicação deste teste obtêm-se populações de formas metacíclicas mais homogéneas isto é, obtêm-se um eficaz isolamento destas formas. Assim sendo, obteve-se neste trabalho, um comprimento médio dos promastigotas metacíclicos de 6,98 m para L. braziliensis, de 5,72 m para L. peruviana e de 7,70 m para L. infantum.It was only in 1984 that Sacks and Perkins proved sequential development of Leishmania promastigotes, from noninfective (procyclic form) to infective (metacyclic form), for promastigotes of L. tropica growing in culture and in the sandfly vector. This process was called metacyclogenesis. Metacyclic forms have been described as small, slender promastigotes with relatively long flagella, occurring in the greatest numbers in vitro in a stationary phase population (Sacks&Perkins, 1984; Franke et al,. 1985; Da Silva&Sacks, 1987; Sacks, 1989; Grimm et al,. 1991; Saraiva et al., 2005). Since infectivity was always strongly correlated with a significant decrease in cell volume, this feature suggests that measuring metacyclics might serve as a suitable parameter to distinguish them from procyclic forms (Grimm et al., 1991; Saraiva et al., 2005; Späth&Beverley, 2001). The in vitro growth in two different media, Schneider’s medium and Grace’s insect medium was study in order to find differences in the kinetics and in number and size of metacyclic forms between species, strains, medium and growth phase. This study involved one strain and one clone L. braziliensis (1794 and LC 2452 cl8, respectively), three strains L. infantum (IMT 151, IMT 260 and IMT 373) and one clone L. peruviana (HR78 cl8). The complement lyses test was used to purify the metacyclic promastigotes (Franke et al., 1985; Puentes et al., 1988; Da Silva&Sacks, 1987, Sacks&Perkins, 1984; Sacks, 1989; Grimm et al., 1991). Kinetic’s growth curves revealed to be different between species, strains of the same species and between the two media. Axenic cultures in Schneider medium had faster growth rate, higher promastigote concentrations, shorter stationary phase and shorter lifetime parasites than in Grace medium, for all the strains studied. These results suggest that the media influence the growth rate and longevity of the strain. However, the media revealed no significant influence (p<0.05) concerning promastigote body length. It was also observed that in Schneider medium the metacyclic population was more homogeneous in comparison with Grace’s where much more mixed populations coexist. To all strains, the higher percentage of metacyclic forms was in the decline growth phase although the peak of the total promastigote forms was in the early stationary. The results of the morphological study of parasites, specifically the analysis of the evolution of the length along its growth curve, showed no significant difference between the mean size of the parasite between Schneider’s and Grace’s medium for the same strain, leading to the conclusion that the medium does not influence the size of promastigotes. The same can not be concluded on the growth phase, which showed significant differences for all strains studied. All strains had average lengths of parasite significantly lower in the stationary phase. Comparing the values of the average length of promastigotes presented by the complement lysis test and those obtained in the kinetics study for each strain and at the same stage of growth, we found that the lowest values were obtained with the complement lysis test. These results suggest that the populations obtained with complement lysis test are more homogeneous for the metacyclic forms. Hereupon, the length obtained for matacyclic forms was of 6,98 m for L. braziliensis, 5,72 m for L. peruviana and 7,70 m for L. infantum

Study of the edible Rumex induratus from Côa Valley in hepaticocarcinoma cells

Hepatocellular carcinoma (HCC) is the most common form of liver cancer (about 90%) which remains a worldwide health challenge due to its incidence growth and yet scarce and not specific treatments.HCC is characterized by a metabolic and oxidative stress which induces a prolonged pathological inflammation and cell damage, and also an evident increase in the production of reactive oxygen species (ROS) which will lead to an increase of cellular lipid peroxidation and of hepatic enzymes. Available treatments are not very effective being usually adapted from other illnesses. Plants are considered very important sources for the discovery of new compounds to prevent and treat diseases (1). There are several natural drug leads presently used in chemotherapy including in HCC. Rumex induratus is native in Iberian Peninsula and spontaneous in northwest Portugal where it is used in local cuisine. Its extracts have significative antioxidant activity (2). This work aimed to add value to this species focused on the evaluation of the antitumoral activity of different extracts. Extractions of Rumex induratus (collected in Vale do Côa, Portugal, between March and May 2021) were made in water, ethanol 80% and ethanol 100% and tested in HCC in vitro models using HepG2, HuH7 and Hep3B cells lines. Metabolic activity, cellular morphology, cellular death and the quantification of reactive oxygen species (ROS) (superoxide anion and hydrogen peroxide) were assayed. A reduction in the metabolic activity was dependent on the extract and its concentration: it was observed that the infusion was more cytotoxic and Hep3B the most sensitive cell line. As for cellular morphology, in HuH7 was observed the condensation of the nucleus after treatment with infusion, and in Hep3B the hydroethanolic extract led to cellular contraction. Cellular death evaluated by the membrane potential revealed that HepG2 and HuH7 suffered from exposure to infusion and hydroethanolic extracts, while Hep3B was not so sensitive. As for ROS quantification, a growth pattern in the concentration of the superoxide anion and a decrease in peroxides were detected with an increase in the concentration of R. induratus extracts. Although not so robust, results of this study still revealed an evident influence of R. induratus extracts in the cellular activity and intracellular ROS production in the studied hepatocellular carcinoma cell lines. It is understandable that these biologic activities are directly associated to the existent phenolic compounds which antioxidant potential allows the understanding that the extracts led to an increase in membrane potential in all cell lines, with HepG2 presenting the highest values. In conclusion, the plant extract provides intracellular differences that can lead to the triggering of apoptosis in tumor cells. Nevertheless, future assays will allow to understand the mechanism of action of R. induratus extracts in HCC and to potentiate its use.info:eu-repo/semantics/publishedVersio

Portugal Doenças Oncológicas em números, 2015

A transformação epidemiológica da Oncologia, tem levado a um crescimento progressivo do número de novos casos anuais, e a um aumento da idade média da população afetada. O aumento de incidência deve- se maioritariamente aos ganhos de esperança de vida da população portuguesa. As modificações dos estilos de vida, para além de influenciarem as variações de incidência, contribuem para mudanças relativas entre as diversas neoplasias. A conjunção destes dois fatores, tem vindo a levar a correções em alta, das previsões de evolução de incidência. Os recursos humanos e materiais necessários tem também crescido significativamente

Portugal Doenças Respiratórias em Números, 2015

A evolução do panorama das Doenças Respiratórias, em Portugal e no Mundo, tem colocado desafios crescentes aos sistemas de saúde, relacionados com uma mudança de paradigma epidemiológico caracterizada por um aumento crescente das doenças respiratórias crónicas (DRC) em oposição a um decréscimo das doenças respiratórias agudas. O aumento do peso das DRC está relacionado não só com os efeitos a curto e longo prazo do tabagismo, mas também com o aumento progressivo da esperança de vida das populações, surgindo as DRC como uma importante causa de morbilidade e mortalidade nas fases avançadas da vida. A mortalidade respiratória atinge sobretudo as faixas etárias acima dos 65 anos de idade, não sendo portanto prematura. Em termos comparativos internacionais, Portugal apresenta um dos melhores valores de taxa padronizada de mortalidade por asma e por Doença Pulmonar Obstrutiva Crónica (DPOC). Contudo, os valores para a taxa de padronizada de mortalidade por pneumonias são um dos piores da Europa

The socio-economic impact of a Polytechnic Institution in a local economy : some insights of field research

Higher Education Institutions (HEI) are considered important mechanisms of regional development. However, due to the current economic recession and budget constraints, HEIs need to demonstrate the impact that their activities have on the surrounding community that contributes for its economic development. Hence, the aim of this paper is to present the results of a model to estimate the economic impact of different HEIs that are located in regions with different socio-economic development. Moreover, the common framework of analysis, whilst considering the different socio-economic realities, allows the comparisons of the results for the different HEIS

EAIR 35th Annual Forum in Rotterdam, the Netherlands 28-31 August 2013

Higher Education Institutions (HEI) are considered important mechanisms of regional development. However, due to the current economic recession and budget constraints, HEIs need to demonstrate the impact that their activities have on the surrounding community that contributes for its economic development. Hence, the aim of this paper is to present the results of a model to estimate the economic impact of different HEIs that are located in regions with different socio-economic development. Moreover, the common framework of analysis, whilst considering the different socio-economic realities, allows the comparisons of the results for the different HEIS.info:eu-repo/semantics/publishedVersio

SARS-CoV-2 introductions and early dynamics of the epidemic in Portugal

Genomic surveillance of SARS-CoV-2 in Portugal was rapidly implemented by the National Institute of Health in the early stages of the COVID-19 epidemic, in collaboration with more than 50 laboratories distributed nationwide. Methods By applying recent phylodynamic models that allow integration of individual-based travel history, we reconstructed and characterized the spatio-temporal dynamics of SARSCoV-2 introductions and early dissemination in Portugal. Results We detected at least 277 independent SARS-CoV-2 introductions, mostly from European countries (namely the United Kingdom, Spain, France, Italy, and Switzerland), which were consistent with the countries with the highest connectivity with Portugal. Although most introductions were estimated to have occurred during early March 2020, it is likely that SARS-CoV-2 was silently circulating in Portugal throughout February, before the first cases were confirmed. Conclusions Here we conclude that the earlier implementation of measures could have minimized the number of introductions and subsequent virus expansion in Portugal. This study lays the foundation for genomic epidemiology of SARS-CoV-2 in Portugal, and highlights the need for systematic and geographically-representative genomic surveillance.We gratefully acknowledge to Sara Hill and Nuno Faria (University of Oxford) and Joshua Quick and Nick Loman (University of Birmingham) for kindly providing us with the initial sets of Artic Network primers for NGS; Rafael Mamede (MRamirez team, IMM, Lisbon) for developing and sharing a bioinformatics script for sequence curation (https://github.com/rfm-targa/BioinfUtils); Philippe Lemey (KU Leuven) for providing guidance on the implementation of the phylodynamic models; Joshua L. Cherry (National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health) for providing guidance with the subsampling strategies; and all authors, originating and submitting laboratories who have contributed genome data on GISAID (https://www.gisaid.org/) on which part of this research is based. The opinions expressed in this article are those of the authors and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. This study is co-funded by Fundação para a Ciência e Tecnologia and Agência de Investigação Clínica e Inovação Biomédica (234_596874175) on behalf of the Research 4 COVID-19 call. Some infrastructural resources used in this study come from the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT).info:eu-repo/semantics/publishedVersio

Multiple effects of toxins isolated from Crotalus durissus terrificus on the hepatitis C virus life cycle

Hepatitis C virus (HCV) is one of the main causes of liver disease and transplantation worldwide. Current therapy is expensive, presents additional side effects and viral resistance has been described. Therefore, studies for developing more efficient antivirals against HCV are needed. Compounds isolated from animal venoms have shown antiviral activity against some viruses such as Dengue virus, Yellow fever virus and Measles virus. In this study, we evaluated the effect of the complex crotoxin (CX) and its subunits crotapotin (CP) and phospholipase A2 (PLA2-CB) isolated from the venom of Crotalus durissus terrificus on HCV life cycle. Huh 7.5 cells were infected with HCVcc JFH-1 strain in the presence or absence of these toxins and virus was titrated by focus formation units assay or by qPCR. Toxins were added to the cells at different time points depending on the stage of virus life cycle to be evaluated. The results showed that treatment with PLA2-CB inhibited HCV entry and replication but no effect on HCV release was observed. CX reduced virus entry and release but not replication. By treating cells with CP, an antiviral effect was observed on HCV release, the only stage inhibited by this compound. Our data demonstrated the multiple antiviral effects of toxins from animal venoms on HCV life cycle

Os principais tipos e manifestações da Cirrose Hepática: uma atualização clínica

Introdução: A cirrose hepática é um processo patológica crônico, considerado a hepatopatia mais comum, definido como a conversão difusa morfoestrutural por nódulos de arquitetura anômalo envoltos por fibrose. Objetivou-se&nbsp;descrever os tipos mais relevantes de cirrose e suas devidas manifestações. Metodologia: Trata-se de uma revisão bibliográfica, fundamentada nas plataformas do SciELO, PubMed, Scopus, utilizando os termos “hepatical cirrhosis”, “liver disease” e “hepatocellular insufficiency” a qual através da revisão narrativa, abordou amplamente a respeito da contextualização da cirrose e as principais etiologias. Resultados e Discussão: Foi analisado que tal condição afeta qualquer faixa etária, sexo, etnia e independe da classe socioeconômica, mas as diversas etiologias impõem um perfil epidemiológico específico conforme a aparição. As principais origens abordam o tipo alcoólico, hepatite, aplicação crônica de alguns fármacos e esteatose gordurosa ou não. Ademais, estima-se que estas afetam a anatomofuncionalidade do órgão responsável por grande parte da homeostase, culminando em diversas manifestações clínicas.&nbsp; Conclusão: A cirrose é uma consequência grave de fatores de base em estágio avançado, a qual devido ao seu curso geralmente silencioso culmina no desenvolvimento e progressão clínica. Neste contexto, a atenção aos fatores predisponentes como alimentação rica em lipídios, estilismo, negligência a exames de rotina, sedentarismo e obesidade contribuem constituem medidas eficazes de prevenção primária.&nbsp