FSR Quorum Sensing: Role in Enterococcus faecalis Biology & Host Infection

Dissertation presented to obtain the Ph.D degree in Biology.

Potentiality of Achatina fulica Bowdich, 1822 (Mollusca: Gastropoda) as intermediate host of the Angiostrongylus costaricensis Morera & Céspedes 1971

Samples of Achatina fulica were experimentally infected with Angiostrongylus costaricensis larvae, etiological agent of abdominal angiostrongyliasis, showing that A. fulica is susceptible to the parasite. Achatina fulica may be a risk to urbanization of abdominal angiostrongyliasis presumably due to its high proliferation, continuous dispersion and remarkable adaptation in several Brazilian towns.

Interaction of malaria parasites with host late endocytic and autophagic pathways is essential for Plasmodium liver stage development

RESUMO: A Malária é causada por parasitas do género Plasmodium, sendo a doença parasitária mais fatal para o ser humano. Apesar de, durante o século passado, o desenvolvimento económico e a implementação de diversas medidas de controlo, tenham permitido erradicar a doença em muitos países, a Malária continua a ser um problema de saúde grave, em particular nos países em desenvolvimento. A Malária é transmitida através da picada de uma fêmea de mosquito do género Anopheles. Durante a picada, os esporozoítos são injetados na pele do hospedeiro, seguindo-se a fase hepática e obrigatória do ciclo de vida. No fígado, os esporozoítos infetam os hepatócitos onde se replicam, dentro de um vacúolo parasitário (VP) e de uma forma imunitária silenciosa, em centenas de merozoitos. Estas novas formas do parasita são as responsáveis por infetar os eritrócitos, iniciando a fase sanguínea da doença, onde se os primeiros sintomas se manifestam, tais como a característica febre cíclica. A fase hepática da doença é a menos estudada e compreendida. Mais ainda, as interações entre o VP e os organelos da células hospedeira estão ainda pouco caracterizados. Assim, neste estudo, as interações entre os organelos endocíticos e autofágicos da célula hospedeira e o VP foram dissecados, observando-se que os anfisomas, que são organelos resultantes da intersecção do dois processos de tráfego intracelular, interagem com o parasita. Descobrimos que a autofagia tem também uma importante função imunitária durante a fase hepática inicial, ao passo, que durante o desenvolvimento do parasita, já numa fase mais tardia, o parasita depende da interação com os endossomas tardios e anfisomas para crescer. Vesiculas de BSA, EGF e LC3, foram, também, observadas dentro do VP, sugerindo que os parasitas são capazes de internalizar material endocítico e autofágico do hospedeiro. Mais ainda, mostramos que esta interação depende da cinase PIKfyve, responsável pela conversão do fosfoinositidio-3-fosfato no fosfoinositidio-3,5-bifosfato, uma vez que inibindo esta cinase o parasita não é capaz de crescer normalmente. Finalmente, mostramos que a proteína TRPML1, uma proteína efetora do fosfoinositidio-3,5-bifosfato, e envolvida no processo de fusão das membranas dos organelos endocíticos e autofágicos, também é necessária para o crescimento do parasita. Desta forma, o nosso estudo sugere que a membrana do VP funde com vesiculas endocíticas e autofágicas tardias, de uma forma dependente do fositidio-3,5-bifosfato e do seu effetor TRPML1, permitindo a troca de material com a célula hospedeira. Concluindo, os nossos resultados evidenciam que o processo autofágico que ocorre na célula hospedeira tem um papel duplo durante a fase hepática da malaria. Enquanto numa fase inicial os hepatócitos usam o processo autofágico como forma de defesa contra o parasita, já durante a fase de replicação o VP funde com vesiculas autofágicas e endocíticas de forma a obter os nutrientes necessários ao seu desenvolvimento.--------- ABSTRACT: Malaria, which is caused by parasites of the genus Plasmodium, is the most deadly parasitic infection in humans. Although economic development and the implementation of control measures during the last century have erradicated the disease from many areas of the world, it remains a serious human health issue, particularly in developing countries. Malaria is transmitted by female mosquitoes of the genus Anopheles. During the mosquito blood meal, Plasmodium spp. sporozoites are injected into the skin dermis of the vertebrate host, followed by an obligatory liver stage. Upon entering the liver, Plasmodium parasites infect hepatocytes and silently replicate inside a host cell-derived parasitophorous vacuole (PV) into thousands of merozoites. These new parasite forms can infect red blood cells initiating the the blood stage of the disease which shows the characteristic febrile malaria episodes. The liver stage is the least characterized step of the malaria infection. Moreover, the interactions between the Plasmodium spp. PV and the host cell trafficking pathways are poorly understood. We dissected the interaction between Plasmodium parasites and the host cell endocytic and autophagic pathways and we found that both pathways intersect and interconnect in the close vicinity of the parasite PV, where amphisomes are formed and accumulate. Interestingly, we observed a clearance function for autophagy in hepatocytes infected with Plasmodium berghei parasites at early infection times, whereas during late liver stage development late endosomes and amphisomes are required for parasite growth. Moreover, we found the presence of internalized BSA, EGF and LC3 inside parasite vacuoles, suggesting that the parasites uptake endocytic and autophagic cargo. Furthermore, we showed that the interaction between the PV and host traffic pathways is dependent on the kinase PIKfyve, which converts the phosphoinositide PI(3)P into PI(3,5)P2, since PIKfyve inhibition caused a reduction in parasite growth. Finally, we showed that the PI(3,5)P2 effector protein TRPML1, which is involved in late endocytic and autophagic membrane fusion, is also required for parasite development. Thus, our studies suggest that the parasite parasitophorous vacuole membrane (PVM) is able to fuse with late endocytic and autophagic vesicles in a PI(3,5)P2- and TRPML1-dependent manner, allowing the exchange of material between the host cell and the parasites, necessary for the rapid development of the latter that is seen during the liver stage of infection. In conclusion, we present evidence supporting a specific and essential dual role of host autophagy during the course of Plasmodium liver infection. Whereas in the initial hours of infection the host cell uses autophagy as a cell survival mechanism to fight the infection, during the replicative phase the PV fuses with host autophagic and endocytic vesicles to obtain nutrients required for parasite growth.

Histoplasmosis presenting as addisonian crisis in an immunocompetent host

A 71-year-old man with presumptively treated pulmonary tuberculosis ten years earlier and previous alcoholism presented with adrenal insufficiency. HIV serology was negative. A computerized tomography scan of the abdomen showed enlarged right adrenal. He recovered after emergency treatment with hydrocortisone IV. Right adrenalectomy was performed. Histoplasmosis was diagnosed and the patient was treated with itraconazole, corticosteroid replacement, and discharged with good health.

Applications of the indole scaffold in medicinal chemistry: development of new antioxidants, COX inhibitors and antitubercular agents

Dissertação para obtenção do Grau de Doutor em Química, especialidade Química Orgânica

Manipulation of host cell functions by Salmonella: the role of the bacterial effector protein SteA

Salmonella enterica serovars are Gram-negative facultative intracellular bacterial pathogens that infect a wide variety of animals. Salmonella infections are common in humans, causing usually typhoid fever and gastrointestinal diseases. Salmonella enterica serovar Typhimurium (S. Typhimurium), which is a leading cause of human gastroenteritis, has been extensively used to study the molecular pathogenesis of Salmonella, because of the availability of sophisticated genetic tools, and of suitable animal and tissue culture models mimicking different aspects of Salmonella infections.(...)

Host genetic factors in mouse malaria liver stage infection

Fundação para a Ciência e a Tecnologia

How much cultural events are "worth" for host communities: Case of Faz Música Lisboa

This paper studies the economic and social impact of Faz Música Lisboa on the segment of society that feels from any direct or indirect consequence due to its existence. A qualitative research based on surveys and interviews is made to retract a list of the benefits and costs that each stakeholder perceives. Relying on the quantifiable variables, it is performed a cost-benefit analysis to measure how much the event is “worth” for the community. I conclude this is a viable project, as it brings a positive net benefit to the society, value that could increase with a higher institutional support.

UK Mental health professionals volunteering in LMIC-benefits to UK and host countries

RESUMO: O Royal College of Psychiatrists tem um programa de voluntariado internacional desde 2005. O interesse no Reino Unido tem crescido nos últimos 15 anos com muitos projetos novos. Os benefícios para vários países têm sido reconhecidos. O que está menos claro são os benefícios da experiência para o Reino Unido e do programa de voluntariado do Royal College of Psychiatrists. O objectivo desta dissertação é explorar os benefícios do voluntariado para o Sistema Nacional de Saúde focando principalmente, mas não exclusivamente, o programa de voluntariado do Royal College of Psychiatrists. Nesta dissertação abordamos primeiro o contexto antes de discutirmos dois grandes estudos de psiquiatras e hospitais do Sistema Nacional de Saúde no Reino Unido. Incluimos no estudo todos os psiquiatras registados como Voluntários no Royal College of Psychiatrists e o Grupo de Interesse Especial associado. Foi ainda possível incluir os Diretores Médicos de todos os hospitais do Reino Unido. Os estudos foram desenhados para analisar as opiniões dos Voluntários do Reino Unido sobre os benefícios para os países recetores de baixo e médio rendimento e para o Reino Unido. Todos os 60 hospitais do Sistema Nacional de Saúde foram incluí dos para analisar a opinião sobre Voluntariado. As limitações dos estudos foram a baixa taxa de resposta, mas esta foi comparável a outros estudos do College. É provável haver um enviezamento de resposta e favorecimento dos que estão envolvidos no Voluntariado. Os resultados mostraram um interesse forte no voluntariado e benefícios no Sistema Nacional de Saúde, nas. áreas de liderança, transculturalismo, gestão e recursos e capacidades pessoais. Os hospitais do Sistema Nacional de Saúde valorizaram o voluntariado e, em contrapartida, o valor acrescido para competências profissionais. Os obstáculos contra o Voluntariado a nível individual e dos hospitais foram maioritariamente de natureza prática, tais como obter dispensa do trabalho, substituição no trabalho e custos. As implicações destes estudos são que o programa de voluntariado do College precisa de ser fortalecido. Para assegurar que este trabalho continua é necessário existir uma sensibilização nacional e maior responsabilização sobre os benefícios para o Reino Unido e a nível global. -------------------------- ABSTRACT: The Royal College of Psychiatrists has had an international volunteering programme since 2005. The interest in UK has grown over the past 15 years with many new projects. The benefits in various countries has been acknowledged. What has been less clear are the benefits of the experience back in the UK and of the Royal College of Psychiatrists Volunteer Scheme. The aim of this dissertation is to explore the benefit of volunteering to the NHS focusing mainly, but not exclusively on the Royal College of Psychiatrists Volunteering scheme. In this dissertation we first look at background information before discussing two large surveys of Psychiatrists and NHS Trusts in UK. We surveyed all those registered as Volunteers at the Royal College of Psychiatrists and the associated Special Interest Group. We also were able to survey the Medical Directors of all UK Trusts. The Surveys were designed to assess views of UK Volunteers of benefits to hosts in LMIC and back in UK. All 60 NHS Trusts were surveyed to assess the view of Volunteering. Limitation of the surveys were the low response rate but this was comparable to other College surveys. There is likely to be a bias in response and favour those who are engaged in the Volunteering agenda. Results showed a strong interest in volunteering and perceived benefits in NHS. These areas included leadership, transcultural, resource management and personal skills. NHS trusts valued volunteering and added value to professional competencies on return. Obstacles to Volunteering at individual and Trust level were mainly practical issues such as getting time off, cover and costs. Implications of these surveys are that the College volunteering scheme needs strengthening. There needs to be a National advocacy to ensure that this work continues and greater accountability as to benefits in UK and globally.

The chemistry of Brazilian leguminosae. LXI. (+)-Methyl Vouacapenate from Vouacapoua pallidior

Thz wood of Vouacapoua pallidior (Leguminosae-Caesalpinioideae) Contains (+)-methyl vouacapenate.

Curare - Botany, Chemistry, and Pharmacology

Some aspects of curare research carried out over the last 25 years are discussed. Accepting a pharmacological rather than purely ethnological definition means, that curares are not limited to South America but that they are also known from Central Africa and South-EastAsia. Among the criteria that have been suggested for classifying South American curares: type of container, geographical origin, botanical sourcesof the active, constituent!, and chemical composition. A combination of botanical and geographical criteria leads to much the same regional ;groupings a combination of criteria involving the type of container and the chemical composition. The active principles in curares may derive from members of thr Loganiaceae (Strychnos) and/or Menispermaceae mainly Chondrodendron and Curarea, but also Abuta,Anomospermum, Cissampelos, Sciadotenia, and Telitoxicum). Certain of the Strychnos dimeric indole alkaloids can undergo a variety of cleavages, oxidations, and isomerizations; hence., some of the compounds obtained by normal isolation procedures one almost certainly artefacts. The different genera of, Menispermaceae a wide range of bisbenzyl and other types of isoquinoline alkaloids. Many of the plant additives also contain a variety of isoquinoline bases, and this has to be taken into account in assessing the contribution these ingredients may make to the ovzJuxll activity of, curare. Loganiaceae-bated curares with toxiferinzas major alkaloid tend to be the most toxic. In the case of Menispermaceae-based products, there-is evidence that the process by which they are made may lead to a considerable increase in the toxicity of the finished poisons as compared with the original plant materials. The mechanism of action of the alkaloids it, outlined, and the role of curare alkaloids in the development of, present-day muscle-relaxant drugs used in surgery is indicated. Attention lb drawn to reported medicinal uses of some of the alkaloid-bearing plants incorporated into curares, suggesting that further evaluation of these plants may be of interest.

The ceramic artifacts in archaeological black earth (terra preta) from Lower Amazon Region, Brazil: chemistry and geochemical evolution

This paper carried out a chemical investigation of archaeological ceramic artifacts found in archaeological sites with Black Earth (ABE) in the Lower Amazon Region at Cachoeira-Porteira, State of Pará, Brazil. The ceramic artifacts, mostly of daily use, belong to Konduri culture (from 900 to 400 years BP). They are constituted of SiO2, Al2O3, Fe2O3, Na2O and P2O5; SiO2 and Al2O3 together add up to 80 % and indicate influence of acid rocks, transformed into clay minerals basically kaolinite. The relative high contents of P2O5 (2.37 % in average) come out as (Al,Fe)-phosphate, an uncommon fact in primitive red ceramics, but found in some roman and egyptian archaeological sites. The contents of the trace elements are similar or below the Earth's crust average. This chemical composition (except P2O5) detaches saprolite material derived acid igneous rocks or sedimentary ones as the main raw material of the ceramics. The contents of K, Na and Ca represent the feldspars and rock fragments possibly introduced into saprolitic groundmass, indicated by mineralogical studies. The presence of cauixi and cariapé as well as quartz sand was confirmed by optical microscope, SEM analyses and by the high silica contents of ceramic fragments. Phosphorus was possibly incorporated into groundmass during cooking of foods, and ABE soil profile formation developed on yellow Latosols. The raw materials and its tempers (cauixi, or cariapé, feldspar, crushed rocks, old ceramic artifacts and quartz fragments) are found close to the sites and therefore and certainly came from them.

Thin films composed of gold nanoparticles dispersed in a dielectric matrix: the influence of the host matrix on the optical and mechanical responses

Gold nanoparticles were dispersed in two different dielectric matrices, TiO2 and Al2O3, using magnetron sputtering and a post-deposition annealing treatment. The main goal of the present work was to study how the two different host dielectric matrices, and the resulting microstructure evolution (including both the nanoparticles and the host matrix itself) promoted by thermal annealing, influenced the physical properties of the films. In particular, the structure and morphology of the nanocomposites were correlated with the optical response of the thin films, namely their localized surface plasmon resonance (LSPR) characteristics. Furthermore, and in order to scan the future application of the two thin film system in different types of sensors (namely biological ones), their functional behaviour (hardness and Young's modulus change) was also evaluated. Despite the similar Au concentrations in both matrices (~ 11 at.%), very different microstructural features were observed, which were found to depend strongly on the annealing temperature. The main structural differences included: (i) the early crystallization of the TiO2 host matrix, while the Al2O3 one remained amorphous up to 800 °C; (ii) different grain size evolution behaviours with the annealing temperature, namely an almost linear increase for the Au:TiO2 system (from 3 to 11 nm), and the approximately constant values observed in the Au:Al2O3 system (4–5 nm). The results from the nanoparticle size distributions were also found to be quite sensitive to the surrounding matrix, suggesting different mechanisms for the nanoparticle growth (particle migration and coalescence dominating in TiO2 and Ostwald ripening in Al2O3). These different clustering behaviours induced different transmittance-LSPR responses and a good mechanical stability, which opens the possibility for future use of these nanocomposite thin film systems in some envisaged applications (e.g. LSPR-biosensors).

Parasite-host relationship between the tambaqui (Colossoma macropomum Cuvier 1818) and ectoparasites, collected from fish farms in the city of Rolim de Moura, State of Rondônia, Western Amazon, Brazil

In this study we investigated the relationship between tambaqui fish (Colossoma macropomumi Cuvier 1818) and parasites in two fish farms (L204S and L180N) in the state of Rondônia, Brazil, during a 1-year period. The objective of the study was to describe the relationships between parasites, hosts and the environment. From the 80 fish specimens collected, 100% were parasitized by at least one parasite species. Seven ectoparasites species were recorded, six of the class Monogenea: Anacanthorus spathulatusi, Mymarothecium spp. (Mymarothecium sp. 1, Mymarothecium sp. 2 and M. viatorum), Notozothecium sp. and Linguadactyloides brinkimanni, classified as dominants, and the copepod Perulernaea gamitanae, classified as subordinate. Despite their high prevalence, the parasites were not abundant. A. spathulatus presented positive and significant correlations between the abundance of parasitism and the standard length of the hosts in the two fish farms; Mymarothecium spp. showed significant correlations, negative in L180N, and positive in L204S; significant positive correlations were observed for Notozothecium sp. in L204S, and for L.brinkimanni in L180N. Young monogeneans were found; these parasites presented a negative correlation in L180N and a significant negative correlation in L204S. The results of the correlation between the relative condition factor (Kn) and the abundance of parasites were not significant for the recorded parasite species. Regarding the hepatosomatic relation (HSR) of fish and the abundance of parasites, Anacanthorus spathulatusi showed a significant negative correlation with the HSR in L180N, and a positive correlation in L204S. Mymarothecium spp. and Notozothecium sp. presented significant positive correlations in L204S. Considering the correlation of the fish splenosomatic relation (SSR) and the abundance of parasites, L. brinkimanni presented significant correlations, positive in L180N and negative in L204S. Despite 100% prevalence, the high water quality contributes to infracommunities with low parasite abundance and good levels of Kn, HSR and SSR, allowing good tambaqui development.