Development of nanohydrogels as a vehicle for the release of bioactive compounds in food matrices

PhD in Chemical and Biological Engineering

In-line rheo-optical microstructural characterization of complex polymer systems

Tese de Doutoramento em Ciência e Engenharia de Polímeros e Compósitos.

The vulnerability to pollution and hydrochemical variation of eleven springs (catchments) in the karst lowlands of the west of Ireland

The vulnerability to pollution and hydrochemical variation of groundwater in the mid-west karstic lowlands of Ireland were investigated from October 1992 to September 1993, as part of an EU STRIDE project at Sligo Regional Technical College. Eleven springs were studied in the three local authority areas of Co. Galway, Co. Mayo, and Co. Roscommon. Nine of the springs drain locally or regionally important karstic aquifers and two drain locally important sand and gravel aquifers. The maximum average daily discharge of any of the springs was 16,000 m3/day. Determination of the vulnerability of groundwater to pollution relies heavily on an examination of subsoil deposits in an area since they can act as a protecting or filtering layer over groundwater. Within aquifers/spring catchments, chemical reactions such as adsorption, solution-precipitation or acid-base reactions occur and modify the hydrochemistry of groundwater (Lloyd and Heathcote, 1985). The hydrochemical processes) that predominate depend cm the mineralogy of the aquifer, the hydrogeological environment, the overlying subsoils, and the history of groundwater movement. The aim of this MSc research thesis was to investigate the hydrochemical variation of spring outflow and to assess the relationship between these variations and the intrinsic vulnerability of the springs and their catchments. If such a relationship can be quantified, then it is hoped that the hydrochemical variation of a spring may indicate the vulnerability of a spring catchment without the need for determining it by field mapping. Such a method would be invaluable to any of the three local authorities since they would be able to prioritise sources that are most at risk from pollution, using simple techniques of chemical sampling, and statistical analysis. For each spring a detailed geological, hydrogeological and hydrochemical study was carried out. Individual catchment areas were determined with a water balance/budget and groundwater tracing. The subsoils geology for each spring catchment were mapped at the 1:10,560 scale and digitised to the 1:25,000 scale with AutoCad™ and Arclnfo™. The vulnerability of each spring was determined using the Geological Survey's vulnerability guidelines. Field measurements and laboratory based chemistry analyses of the springs were undertaken by personnel from both the EPA Regional Laboratory in Castlebar, Co. Mayo, and the Environment Section of Roscommon Co. Council. Electrical conductivity and temperature (°C) were sampled fortnightly, in the field, using a WTW microprocessor conductivity meter. A percentage (%) vulnerability was applied to each spring in order to indicate the areal extent of the four main classes of vulnerability (Extreme, High, Moderate, and Low) which occurred within the confines of each spring catchment. Hydrochemical variation for the springs were presented as the coefficient of variation of electrical conductivity. The results of this study show that a clear relationship exists between the degree of vulnerability of each catchment area as defined by the subsoil cover and the coefficient of variation of EC, with the coefficient of variation increasing as the vulnerability increases. The coefficient of variation of electrical conductivity is considered to be a parameter that gives a good general reflection of the degree of vulnerability occurring in a spring catchment in Ireland's karstic lowlands.

Population biology of the crab Armases angustipes (Crustacea, Decapoda, Sesarmidae) at Brazilian tropical coast

The semi terrestrial crabs are important elements of the fauna of coastal regions. The aim of this study was to analyze the population structure of Armases angustipes (Dana, 1852) at estuaries of the Ariquindá River, considered a non impacted area, and Mamucabas River, considered a few impacted area, on the south coast of state of Pernambuco, Brazil. The species occurred in all months of the year. The number of individuals per month varied, being higher in the months of transition between the seasons. This is probably due to significant seasonal variations of air and burrow temperature and burrow salinity. There was no sexual dimorphism in size of A. angustipes in the mangrove of Ariquindá River, but males were larger than females in the mangrove of Mamucabas River. In both estuaries, the sex ratio did not differ from Mendelian proportion, but showed a deviation for females. The analysis of temporal variation in sex ratio showed significant differences in some months of the year. These variations are due to cyclical events that act distinctly on each sex. In both estuaries, size classes of carapace width were equally represented by both sexes. The ovigerous females of A. angustipes occurred only in some months of the year, especially in summer, in both estuaries. Probably the high phytoplankton productivity observed in summer favors the reproductive activity, since these algae serve as food for the larvae. Specimens of the population of Rio Ariquindá are largest and wider than those of Mamucabas River. This fact, associated with the low abundance of crabs and the lower frequency of ovigerous females observed in Mamucabas River, is an indication that this population may be influenced by the environmental impacts that this estuary has received.

Manipulation of optimal matchings via predonation of endowment

In this paper we answer a question posed by Sertel and Özkal-Sanver (2002) on the manipulability of optimal matching rules in matching problems with endowments. We characterize the classes of consumption rules under which optimal matching rules can be manipulated via predonation of endowment.

Semigroups of matrices of intermediate growth

Finitely generated linear semigroups over a field K that have intermediate growth are considered. New classes of such semigroups are found and a conjecture on the equivalence of the subexponential growth of a finitely generated linear semigroup S and the nonexistence of free noncommutative subsemigroups in S, or equivalently the existence of a nontrivial identity satisfied in S, is stated. This ‘growth alternative’ conjecture is proved for linear semigroups of degree 2, 3 or 4. Certain results supporting the general conjecture are obtained. As the main tool, a new combinatorial property of groups is introduced and studied.

Differentation of the fat body cells in the precocius adults of Schistocerca gregaria: regulation of polyploidy by juvenile hormone

Precocious adults from 2nd and 3rd instar larvae of the desert locust Schistocerca gregaria were used to assess the competence of their fat body to synthesize DNA in response to a juvenile hormone analog (JHA), hydoprene. Autoradiographic studies show that JHA stimulates DNA synthesis since a significant proportion of the fat body nuclei are labelled after treatment with 100 or 200 µg of JHA. Maximum DNA synthesis occurs 24 h after treatment with 100 µg of JHA. The nuclear ploidy classes of the precocious adults from 3rd larvae are similar to those of 1-d-old normal adults, but treatemnt of these precociuos adults with µg of JHA doubles the DNA content resulting in enhanced ploidy classes which resemble those of 10-d-old normal females. In the precocious adults that emerged from 2nd instar larvae the ploidy classes are higher than those of 1-d-old normal adults, and treatment of these precocious adults with JHA results in a further increase in the DNA content of the fat body nuclei leading to the formation of high percentages of 16C and 32C nuclei. The results of these studies suggest that any model on the mode of action of JH should recognize this phenomenon of JH-induced polyploidization in the fat body nuclei.

The roles of haemolymphatic lipoproteins in the oogenesis of Rhodnius prolixus

The fates of purified 32P-vitellin and 32P-lipophorin were followed in vitellogenic females of Rhodnius prolixus. While the radioactivity from 32P-vitellin 6 hours after injection was found almost exclusively in the ovary, the radioactivity from injected 32P-lipophorin was found distributed among several organs. In the ovary, the radioactivity from 32P-vitellin was associated with the contents of the yolk granules. 32P-lipophorin delivered a great amount of radioactive phospholipids to the ovary with no accumulation of its protein moiety, as observed after its iodination with 131I. The delivery of phospholipids was inhibited at 0ºC and by the metabolic inhibitors, sodium azide and sodium fluoride. Comparison of the radioactivity incorporation from 32P-lipophorin with that of 14C-inulin suggests that the 32P-phospholipids from lipophorin are not taken up by fluid phase endocytosis. The data presented here are compatible with the concept of lipophorin as a carrier of lipids in insects and provide evidence that lipophorin transports phospholipids as shown previously for other classes of lipids. The utilization by the oocytes of the phospholipids transported by lipophorin is discussed.

Epidemiological traits of the malaria-like parasite Polychromophilus murinus in the Daubenton¿s bat Myotis daubentonii.

BackgroundThe great diversity of bat haemosporidians is being uncovered with the help of molecular tools. Yet most of these studies provide only snapshots in time of the parasites discovered. Polychromophilus murinus, a malaria-like blood parasite, specialised on temperate-zone bats is a species that is being `rediscovered¿. This study describes the infection dynamics over time and between host sex and age classes.MethodsFor three years we followed the members of three breeding colonies of Myotis daubentonii in Western Switzerland and screened them for the prevalence and parasitemia of P. murinus using both molecular tools and traditional microscopy. In order to identify more susceptible classes of hosts, we measured, sexed and aged all individuals. During one year, we additionally measured body temperature and haematocrit values.ResultsJuvenile bats demonstrated much higher parasitemia than any other age class sampled, suggesting that first exposure to the parasite is very early in life during which infections are also at their most intense. Moreover, in subadults there was a clear negative correlation between body condition and intensity of infection, whereas a weak positive correlation was observed in adults. Neither body temperature, nor haematocrit, two proxies used for pathology, could be linked to intensities of infection.ConclusionIf both weaker condition and younger age are associated with higher infection intensity, then the highest selection pressure exerted by P. murinus should be at the juvenile stage. Confusion over the identities and nomenclature of malarial-like parasites requires that molecular barcodes are coupled to accurate morphological descriptions.

Monoclonal antibodies to murine lipopolysaccharide (LPS)-binding protein (LBP) protect mice from lethal endotoxemia by blocking either the binding of LPS to LBP or the presentation of LPS/LBP complexes to CD14.

Cellular responses to LPS, the major lipid component of the outer membrane of Gram-negative bacteria, are enhanced markedly by the LPS-binding protein (LBP), a plasma protein that transfers LPS to the cell surface CD14 present on cells of the myeloid lineage. LBP has been shown previously to potentiate the host response to LPS. However, experiments performed in mice with a disruption of the LBP gene have yielded discordant results. Whereas one study showed that LBP knockout mice were resistant to endotoxemia, another study did not confirm an important role for LBP in the response of mice challenged in vivo with low doses of LPS. Consequently, we generated rat mAbs to murine LBP to investigate further the contribution of LBP in experimental endotoxemia. Three classes of mAbs were obtained. Class 1 mAbs blocked the binding of LPS to LBP; class 2 mAbs blocked the binding of LPS/LBP complexes to CD14; class 3 mAbs bound LBP but did not suppress LBP activity. In vivo, class 1 and class 2 mAbs suppressed LPS-induced TNF production and protected mice from lethal endotoxemia. These results show that the neutralization of LBP accomplished by blocking either the binding of LPS to LBP or the binding of LPS/LBP complexes to CD14 protects the host from LPS-induced toxicity, confirming that LBP is a critical component of innate immunity.

Algebraic properties of isochronous centers of polynomial quadratic-like Hamiltonian systems

Projecte de recerca elaborat a partir d’una estada a la School of Mathematics and Statistics de la University of Plymouth, United Kingdom, entre abril juliol del 2007.Aquesta investigació és encara oberta i la memòria que presento constitueix un informe de la recerca que estem duent a terme actualment. En aquesta nota estudiem els centres isòcrons dels sistemes Hamiltonians analítics, parant especial atenció en el cas polinomial. Ens centrem en els anomenats quadratic-like Hamiltonian systems. Diverses propietats dels centres isòcrons d'aquest tipus de sistemes van ser donades a [A. Cima, F. Mañosas and J. Villadelprat, Isochronicity for several classes of Hamiltonian systems, J. Di®erential Equations 157 (1999) 373{413]. Aquell article estava centrat principalment en el cas en que A; B i C fossin funcions analítiques. El nostre objectiu amb l'estudi que estem duent a terme és investigar el cas en el que aquestes funcions són polinomis. En aquesta nota formulem una conjectura concreta sobre les propietats algebraiques que venen forçades per la isocronia del centre i provem alguns resultats parcials.

Stent retriever use for retrieval of displaced microcoils: a consecutive case series.

SUMMARY: Coil displacement during endovascular coiling procedures may require coil retrieval in the context of flow limitation or thromboembolic risk. No standard recommended method of coil retrieval exists. We present a consecutive series of 14 patients with displaced coil during aneurysm coiling in whom the complication was effectively managed with the use of a stent retriever system. Two illustrative cases from the 14 are described, and technical notes are detailed regarding use of the technique. The use of stent retrievers presents a simple, safe, and effective choice for removal of prolapsed coils during aneurysm coiling.

(Optimal) Spatial Aggregation in the Determinants of Industrial Location

Empirical studies on the determinants of industrial location typically use variables measured at the available administrative level (municipalities, counties, etc.). However, this amounts to assuming that the effects these determinants may have on the location process do not extent beyond the geographical limits of the selected site. We address the validity of this assumption by comparing results from standard count data models with those obtained by calculating the geographical scope of the spatially varying explanatory variables using a wide range of distances and alternative spatial autocorrelation measures. Our results reject the usual practice of using administrative records as covariates without making some kind of spatial correction. Keywords: industrial location, count data models, spatial statistics JEL classification: C25, C52, R11, R30

Role of penicillin-binding proteins in Streptococcus gordonii

Résumé Streptococcus gordonii est une bactérie colonisatrice naturelle de la cavité buccale de l'homme. Bien que normalement commensale, elle peut causer des infections graves, telles que des bactériémies ou des endocardites infectieuses. La pénicilline étant un des traitements privilégiés dans de tels cas, l'augmentation rapide et globale des résistances à cet antibiotique devient inquiétante. L'étude de la physiologie et des bases génétiques de ces résistances chez S. gordonii s'avère donc importante. Les cibles moléculaires privilégiées de la pénicilline G et des β-lactames sont les penicilllin-binding proteins (PBPs). Ces enzymes associées à la membrane ont pour rôle de catalyser les réactions de transpeptidation et de transglycosylation, qui constituent les dernières étapes de la biosynthèse du peptidoglycan (PG). Elles sont définies comme classe A ou B selon leur capacité d'assurer soit les deux réactions, soit uniquement la transpeptidation. Les β-lactames inhibent le domaine transpeptidase de toutes les PBPs, entraînant l'inhibition de la synthèse du PG, l'inhibition de la croissance, et finalement la mort cellulaire. Chez les streptocoques, les PBPs sont aussi les premiers déterminants de la résistance à la pénicilline. De plus, elles sont impliquées dans la morphologie bactérienne, en raison de leur rôle crucial dans la formation du PG. Le but de ce travail était de caractériser les PBPs de S. gordonii et d'étudier leurs fonctions dans la vie végétative de la bactérie ainsi que durant le développement de la résistance à la pénicilline. Premièrement, des mutants auxquels il manque une ou deux PBP(s) ont été construits. Leur étude - au niveau physiologique, biochimique et morphologique - a montré le caractère essentiel ou dispensable de chaque protéine, ainsi que certaines de leurs fonctions potentielles. Deuxièmement, des mutants résistants à la pénicilline ont été générés. Leur caractérisation a montré l'importance des mutations dans les PBPs ainsi que dans d'autres gènes encore inconnus, de même que le rôle crucial des PBPs de classe A dans le développement de la résistance à la pénicilline. Des expériences supplémentaires sur des isolats résistants ont aussi prouvé que la résistance a un coût en terme de fitness, coût que S. gordonii parvient à compenser par des mécanismes d'adaptation. Finalement, les promoteurs des gènes des PBPs ont été déterminés et leur expression a été étudiée grâce au gène de luciférase. Il a ainsi été montré que la résistance à la pénicilline entraîne non seulement des altérations au niveau des protéines, mais aussi au niveau de la régulation des gènes. De plus, la pénicilline génère directement des modifications dans l'expression de PBPs spécifiques. Summary Streptococcus gordonii is a normal inhabitant of the human oral cavity and a pioneer colonizer of teeth. Although usually considered as a commensal, this organism can cause life-threatening infections such as bacteraemia or endocarditis. Since penicillin is one of the preferential treatments for such pathologies, the rapid and general increase of antibiotic resistance in the overall population becomes an issue. Thus, studying the physiologic and genetic bases of such a resistance in S. gordonii is of interest. The primary molecular targets of penicillin G and other β-lactams are the so called penicillin-binding proteins (PBPs). These are membrane-associated proteins that catalyze the last steps in peptidoglycan (PG) biosynthesis, namely transpeptidation and transglycosylation. Depending on their capacity to catalyze either reactions or only transpeptidation, they are considered as class A or class B PBPs, respectively. β-lactam antibiotics inhibit the transpeptidase domain of both of these classes of enzymes, resulting in inhibition of PG assembly, inhibition of bacterial growth, and ultimately leading to cell death. In streptococci, PBPs are also the primary determinants of penicillin-resistance. Moreover, because of their crucial role in PG formation, they are implicated in fundamental aspects of cell morphology. The goal of this work was thus to characterize S. gordonii PBPs and to explore their functions in terms of vegetative life and penicillin-resistance development. First, single and double PBP-inactivated mutants were generated and their effect on the bacterial physiology, cell wall biochemistry and ultrastructural morphology was assessed. This demonstrated the essentiality or dispensability of each protein for bacterial life. Second, penicillin-resistant mutants were generated by cyclic exposure to increasing concentrations of the drug. Characterization of these mutants pointed out the importance of both PBP and non-PBP mutations, as well as the crucial role of the class A PBPs in the development of penicillin-resistance. Further experiments on resistant isolates demonstrated the fitness cost of this resistance, but also the capacity of S. gordonii to adapt and regain the fitness of the wild-type. Finally, the promoters of PBP genes were determined and their expression was monitored using luciferase fusions. This showed that penicillin-resistance, in addition to modifications at the level of the protein, also triggered genetic alterations. Moreover, penicillin itself generated modifications in the expression of specific PBPs.

Duplex Hemodinamic evaluation of hepatosplenic mansoni schistosomiasis

Conventional ultrasonography highly contributes to a non invasive diagnosis of HS schistosomiasis (Cerri et al., 1984). The introduction of Dopple allowed new advances in the knowledge of the portal dinamics of this disease (Taylor et al., 1985; Moriyasu et al., 1986). The aim of this paper was to analize the hemodinamic behavior of the portal vessels, considering caliper, maximum flow speed, direction of flow and preferential disposition of the collateral vessels. Thirty two patients with schistosomiasis mansoni with confirmed hepatosplenic form (HSSM), were analyzed. Fourteen patients with the intestinal form, have been analyzed as a control group. The results demonstrated that the maximum speed of the portal vein in the two groups has not been significantly diferent. Nevertheless, the diameter of the PV in the hepatosplenic group has been larger. The splenic vein presented speed and caliper larger than the superior mesenteric vein. The hepatic artery has been detectly in only 40% of the cases. The hepatic veins presented normal caliper and spectral pattern. The duplex proved to be an useful technich complementar and non-invasive, in the study of the HSSM.