Bioinspired polyethersulfone-based hollow fiber membranes as the scaffolds in renal assist device for protein-bound toxins removal from blood

Dissertation for obtaining the Master degree in Membrane Engineering

Castor fiber na gruta do Caldeirão. Existência, distribuição e extinção do castor em Portugal

Beaver only had been found in Portugal in a Chalcolithic locality, the Vila Nova de S. Pedro castrum. It has now been identified in the Upper Paleolithic (Solutrean) from Gruta do Caldeirão, near Tomar. The species has been found recently at «Gruta do Almonda»; 4 teeth were collected in bed C, older than a Solutrean sequence (see Anexo for details). The species seems to have been rare, as it was also the case with portuguese Miocene Castoridae Enroxenomys minutus and Chalicomys jaegeri. If account is taken of the presence in the Middle Ages until Castille of words meaning beaver (relared to the popular latin Fiber/Biber), it is obvious that these animais still existed then. Such nouns were largely predominant over rhe rather erudite latin (greek deríved) words as Castor,-óris and derived ones, as it could be expected. This allowed us to recognize that veiro should be the corresponding word with Fiber affinities in archaic portuguese. It was previously supposed to mean only expensive furs then imported into Portugal. Indeed it was also a zoonym. Anywày, beaver should be scarce by XIIIth century since it is not included in the quite detailed price list imposed by the «Lei da Almotaçaria» from December 26, 1253 (see Quadro II). Toponyms in veiro and derived words (fig. 2; Quadro III) (plural, feminines, diminutives, inhabited places) give a resrrictive view of rhe Middle Age distribution. Some of them are certainly older than Portugal itself (firsr half of XIlth cenrury); others existed by the XIVth century bur were probably older. Some rare toponyms seem to be derived from the erudite latin Castor,-óris. Nothing suggests that these words were still in use as zoonyms during the Middle Ages. All toponyms are located in regions near rivers and other freshwaters ecologically suitable for beavers, so we can approximately retrace its former, Middle Age disrribution in Portugal (fig. 2; Quadro III). Most of them are locared in the Center-West and Northwest of Portugal, with a suitable c1imate (rainfall in general over 800 mílimerers per year); the only sure geographical exception is Veiros, in Alto Alentejo province, in a region with comparable precipitations and less dry climate conditions than mosr of the territories South of rhe Tagus. There are less and less of these toponyms towards rhe South and the inner part of the country, and they are enrirely lacking in all drier regions from Trás-os-Montes, Beira, Alentejo beyond Tagus' basin, and in Algarve. Nothing suggests beavers lived there, No post-medieval toponym is known, nor any reference after middle XVth century. No such locality was at, or close by to, any frontier. Hence the hypothesis of veiro (et al.} as meaning but points where expensive furs (supposedly known as veiros in general but without c1early saying from what animal they were obrained from) is to be discarded. During the Middle Ages, beaver discriburion concerned all the main river basins from Minho to Tagus ones. Quite rarefied in rhe XIIIth, the beavers may have disappeared from Portugal during the XVth century. Ecological requiremenrs restricted their former distriburion. Vulnerability to natural causes (i.e., severe drought) and to human pressure may have accounted heavily for this species' extinction. Last (1446) reference for Portugal known to us suggests the species was by then almost extinct.

High-performance flexible hybrid field-effect transistors based on cellulose fiber paper

IEEE Electron Device Letters, VOL. 29, NO. 9,

Castor fiber na gruta do Caldeirão Existência, distribuição e extinção do castor em Portugal

Beaver only had been found in Portugal in a Chalcolithic locality, the Vila Nova de S. Pedro castrum. It has now been idenrified in the Upper Paleolithic (Solurrean) from Gruta do Caldeirão, near Tomar. The species has been found recently at «Gruta do Almonda»; 4 teeth were collected in bed C, older than a Solutrean sequence (see Anexo for details). The species seems to have been rare, as it was also the case with portuguese miocene Castoridae Enroxenomys minutus and Chalicomys jaegeri. If account is taken of the presence in the Middle Ages until Castille of words meaning beaver (related to the popular latin Fiber/Biber), it is obvious that these animals still existed then. Such nouns were largely predominant over the rather erudite larin (greek derived) words as Castor, -óris and derived ones, as it could be expected. This allowed us to recognize that veiro should be the corresponding word with Fiber affinities in archaic portuguese. It was previously supposed to mean only expensive furs then imported into Portugal. Indeed it was also a zoonym. Anyway, beaver should be scarce by XIIIth century since it is not included in the quite detailed price list imposed by the «Lei da Almotaçaria» from December 26, 1253 (see Quadro II). Toponyms in veiro and derived words (fig. 2; Quadro III) (plural, feminines, diminutives, inhabited places) give a restrictive view of the Middle Age distribution. Some of them are certainly older than Portugal itself (first half of XIIth century); others existed by the XIVth century but were probably older. Some rare toponyms seem to be derived from rhe erudite latin Castor, -óris. Nothing suggests that these words were still in use as zoonyms during the Middle Ages. All toponyms are located in regions near rivers and other freshwaters ecologically suitable for beavers, so wecan approximately retrace irs former, Middle Age distribution in Portugal (fig. 2; Quadro III). Most of them are located in the Center-West and Northwest of Portugal, with a suitable c1imate (rainfall in general over 800 milimeters per year); the only sure geographical exception is Veiros, in Alto Alentejo province, in a region with comparable precipitations and less dry climare conditions than most of the territories South of the Tagus. There are less and less of these toponyms towards the South and the inner part of the country, and they are enrirely lacking in ali drier regions from Trás-os-Montes, Beira, Alentejo beyond Tagus' basin, and in Algarve. Nothing suggests beavers lived there, No pose-medieval toponym is known, nor any reference after middle XVth century. No such locality was at, or close by to, any frontier. Hence the hypothesis of veiro (e: al.) as meaning but points where expensive furs(supposedly known as veiros in general but without clearly saying from what animal they were obtained from) is to be discarded. During the Middle Ages, beaver distribution concerned all the main river basins from Minho to Tagus ones. Quice racefied in the XIIIth, the beavers may have disappeared from Portugal during the XVth century. Ecological requirements restricted their former distribution. Vulnerability to natural causes (i.e., severe drought) and to human pressure may have accounted heavily for this species extinction. Last (1446) reference for Portugal known to us suggests the species was by then almost extinct.

Process engineering of liver cells for drug testing applications

Dissertação para obtenção do Grau de Doutor em Bioengenharia

Dissecting the molecular interaction between hepatocytes and Plasmodium liver parasites

Dissertation presented to obtain the Ph.D degree in Biology

NADH oxidase activity of rat and human liver xanthine oxidoreductase: potential role in superoxide production

J Biol Inorg Chem (2007) 12:777–787 DOI 10.1007/s00775-007-0229-7

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.

Fiber functionalization for biomedical applications

Deep-eutectic solvents (DES) are considered novel renewable and biodegradable solvents, with a cheap and easy synthesis, without waste production. Later it was discovered a new subclass of DES that even can be biocompatible, since their synthesis uses primary metabolites such as amino acids, organic acids and sugars, from organisms. This subclass was named natural deep-eutectic solvents (NADES). Due to their properties it was tried to study the interaction between these solvents and biopolymers, in order to produce functionalized fibers for biomedical applications. In this way, fibers were produced by using the electrospinning technique. However, it was first necessary to study some physical properties of NADES, as well as the influence of water in their properties. It has been concluded that the water has a high influence on NADES properties, which can be seen on the results obtained from the rheology and viscosity studies. The fluid dynamics had changed, as well as the viscosity. Afterwards, it was tested the viability of using a starch blend. First it was tested the dissolution of these biopolymers into NADES, in order to study the viability of their application in electrospinning. However the results obtained were not satisfactory, since the starch polymers studied did not presented any dissolution in any NADES, or even in organic solvents. In this way it was changed the approach, and it was used other biocompatible polymers. Poly(ethylene oxide), poly(vinyl alcohol) and gelatin were the others biopolymers tested for the electrospinning, with NADES. All polymers show good results, since it was possible to obtain fibers. However for gelatin it was used only eutectic mixtures, containing active pharmaceutical ingredients (API’s), instead of NADES. For this case it was used mandelic acid (antimicrobial properties), choline chloride, ibuprofen (anti-inflammatory properties) and menthol (analgesic properties). The polymers and the produced fibers were characterized by scanning electron microscope (SEM), Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR). With the help of these techniques it was possible to conclude that it was possible to encapsulate NADES within the fibers. Rheology it was also study for poly(ethylene oxide) and poly(vinyl alcohol), in a way to understand the influence of polymer concentration, on the electrospinning technique. For the gelatin, among the characterization techniques, it was also performed cytotoxicity and drug release studies. The gelatin membranes did not show any toxicity for the cells, since their viability was maintained. Regarding the controlled release profile experiment no conclusion could be drawn from the experiments, due to the rapid and complete dissolution of the gelatin in the buffer solution. However it was possible to quantify the mixture of choline chloride with mandelic acid, allowing thus to complete, and confirm, the information already obtained for the others characterization technique.

the liver endoplasmic reticulum in malaria liver infection

Malaria, a disease caused by Plasmodium, represents a major health problem with a still disconcertingly high mortality rate (655 000 malaria deaths were estimated by the World Health Organization in 2012), mainly in Africa [1]. After a bite by an infected Anopheles mosquito occurs, Plasmodium sporozoites reach their target organ, the liver, within minutes. After traversing several hepatocytes, the parasite invades a final one and establishes a parasitophorous vacuole, where it replicates exponentially generating thousands of infective merozoites, the red blood cell infectious forms that are released in the blood stream. The liver stage is the first obligatory phase of malaria infection and, although no symptoms are associated with it, it is absolutely crucial to the establishment of a successful infection.(...)

Host genetic factors in mouse malaria liver stage infection

Fundação para a Ciência e a Tecnologia

Dynamics of serotonergic neurons revealed by fiber photometry

This work was developed in the context of the MIT Portugal Program, area of Bioengineering Systems, in collaboration with the Champalimaud Research Programme, Champalimaud Center for the Unknown, Lisbon, Portugal. The project entitled Dynamics of serotonergic neurons revealed by fiber photometry was carried out at Instituto Gulbenkian de Ciência, Oeiras, Portugal and at the Champalimaud Research Programme, Champalimaud Center for the Unknown, Lisbon, Portugal