Teses / dissertações sobre o tema "Metabolism"

Orientador: Maria Terezinha Serrão Peraçoli
Resumo: O aumento da atividade dos radicais livres e da produção de citocinas pró-inflamatórias por monócitos pode estar envolvido na patogênese da pré-eclâmpsia. A silibinina é o componente mais ativo da silimarina (Silybum marianum), um flavonóide polifenólico que possui efeitos anti-oxidante e anti-inflamatório. O objetivo do presente estudo foi avaliar o efeito da silibinina sobre a produção de citocinas pró e anti-inflamatórias, bem como sobre a liberação de ânion superóxido (O2 -) por monócitos de gestantes com pré-eclâmpsia.Foram selecionadas 30 gestantes com préeclâmpsia (PE), 30 gestantes normais (GN) e 30 mulheres normais nãográvidas (NG). Monócitos de sangue periférico foram incubados na presença ou ausência de silibinina nas concentrações de 5 e 50 g/mL por 60min e, estimulados ou não com ester de forbol (PMA) para determinação de O2 -. Estes monócitos também foram estimulados ou não com lipopolissacáride (LPS) por 18h e, no sobrenadante das culturas foram determinadas, pela técnica de ELISA, as seguintes citocinas: fator de necrose tumoral-alfa (TNF- ), Interleucinas (IL-) IL-6, IL-10, IL-12, IL-15, fator estimulador de colônias de granulócitos e monócitos (GM-CSF) e fator de crescimento e transformação (TGF- . A atividade da enzima superóxido dismutase (SOD) foi avaliada em hemolisado de eritrócitos dos três grupos estudados.Níveis significativamente maiores de O2 - foram liberados por monócitos de gestantes com PE quando comparados com GN e NG, confirmando o estado ativado dessas células. A atividade da SOD foi significativamente maior no grupo de gestantes com PE. Os níveis endógenos de TNFforam significativamente mais elevados nas pacientes com PE, quando comparados com os grupos GN e NG, enquanto os níveis de IL-10 foram significativamente menores nas gestantes com PE. A capacidade de produção de citocinas pelos... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Increased free radical activity and high proinflammatory cytokine production by monocytes may be implicated in the pathogenesis of preeclampsia. Silibinin is a major active component of silymarin (Silybum marianum), a polyphenolic plant flavonoid that has antioxidant and anti-inflammatory effects. This study investigated the in vitro effect of silibinin on monocyte production of pro- and anti-inflammatory cytokines, as well as on superoxide anion (O2-) release in pregnant women with preeclampsia.Thirty preeclamptic (PE), 30 healthy pregnant (HP) and 30 healthy non-pregnant (NP) women were included. Peripheral blood monocytes were incubated with or without silibinin at 5 and 50ug/mL for 60 min, and stimulated with or without phorbol myristate acetate (PMA) for the assessment of O2-. These cells were also cultured with or without lipopolysaccharide (LPS) for 18h and tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-6, IL-10, IL-12p40, IL-15, granulocyte-macrophage colony-stimulating factor (GM-CSF) and transforming growth factor-beta1 (TGF- 1) in monocyte culture supernatants were determined by ELISA. The activity of the enzyme superoxide dismutase (SOD) was evaluated in erythrocyte lisates of the three groups studied.Monocytes of preeclamptic patients release significantly higher levels of O2 - in comparison to HP and NP women confirming the activation state of these cells. SOD activity in erythrocytes was significantly higher in preeclamptic patients. The endogenous levels of TNFwere significantly higher in PE patients than in HP and NP groups, while IL-10 production was significantly lower in PE women. The levels of IL-6, IL-12 and GM-CSF spontaneously produced by monocytes were higher in HP and PE groups than in NP women. The capacity of cytokine production by LPS-stimulated monocytes was preserved in all the three groups studied... (Complete abstract click electronic access below)
Mestre

La línea de investigación presentada en esta tesis representa un primer acercamiento entre los estudios sobre Hidrología y Metabolismo Social. La línea nace de la observación de que el agua es evitada en los estudios que tratan el metabolismo y de que la ciencia del agua –si bien reconoce la necesidad de evolucionar hacia la interdisciplinariedad- todavía no ha conseguido conectar los análisis enfocados en la sociedad y en los ecosistemas. La contribución que se hace en este trabajo es precisamente la definición de un marco analítico –el Metabolismo Hídrico de los Socio-ecosistemas- donde se puede establecer esta conexión y que está formado por una propuesta conceptual y un set de herramientas metodológicas. El documento se divide en tres partes donde se discuten las novedades epistemológicas, metodológicas y formales del marco. La Parte I cubre las reflexiones epistemológicas relacionadas con el marco analítico. Éstas comienzan en el Capítulo 1 con la explicación de los restos a los que la ciencia del agua se enfrenta y que están relacionados con la necesidad de encontrar marcos analíticos que puedan proporcionar inputs relevantes para la gestión integrada de los recursos hídricos (GIRH). Al igual que para el caso de otros recursos, la GIRH requiere el establecimiento de una conexión analítica de las dinámicas sociales y de los ecosistemas. La analogía del metabolismo de la sociedad, como una de las piezas claves de la Ciencia de la Sostenibilidad, es una buena opción para establecer esta conexión. Sin embargo, el concepto de metabolismo necesita ser examinado de cerca antes de su uso combinado con otras concepciones de las relaciones entre el ser humanos y la naturaleza. Tras subrayas que el metabolismo de las sociedades y los ecosistemas son dos procesos distintos per conectados, en el Capítulo 2 se propone un esquema para la descripción de las relaciones metabólicas entre ellos. En el capítulo 3, este esquema es adaptado a las condiciones específicas del agua, usando algunos de los conceptos más relevantes en socio- y eco-hidrología. De esta forma, el metabolismo hídrico del socio-ecosistema es definido como el metabolismo del sistema agua-ser humano. La Parte II describe el marco metodológico. Como un marco ampliamente establecido que es capaz de tratar los problemas de escala y de integrar narrativas, el Capítulo 4 presenta el Análisis Integrado Multi-Escala del Metabolismo Social y de los Ecosistemas (MuSIASEM). MuSIASEM se ha seleccionado como raíz y ha sido adaptado al análisis de sistemas complejos agua-ser humano. Dado que el agua presenta importantes diferencias con respecto a los análisis previos en energía, esta adaptación requiere la inclusión de nuevas escalas de análisis –el ‘problemshed’ y el ‘watershed’- y nuevas definiciones del agua como metabolito –como flujo y fondo. En el capítulo 5 se señalan las diferencias y sinergias ente MuSIASEM el análisis de la huella hídrica –como una de las herramientas de la GIRH. En la Parte III se presentan cuatro casos de estudio con dos objetivos. En primer lugar, el Capítulo 6 analiza a sostenibilidad de los patrones metabólicos en el uso del agua en Punjab y Mauricio para testear la aplicación de MuSIASEM a los estudios de agua y para mostrar cómo este tipo de análisis de formaliza. En segundo lugar, el Capítulo 7 muestra como los métodos de contabilidad del agua del análisis de la huella hídrica complementan el análisis de flujos de agua en MuSIASEM, encontrando además una referencia para su contextualización.
The research line presented in this dissertation is a first attempt to provide a bridge for the communication between Hydrological studies and Social Metabolism. It was born from the observation that water is neglected in Social Metabolism and that current water science, while certain about the need of evolving towards a more interdisciplinary field, still faces challenges in the connection of social and ecosystem analyses. The contribution made here is the definition of an analytical framework –the Water Metabolism of Socioecosystems- where this connection can be established and which is formed by a conceptual proposal and a methodological toolkit. The document is divided in three parts where the epistemological, the methodological and the formal novelties of the framework are discussed. Part I covers the epistemological reflections related to the analytical framework. It begins in Chapter 1 with the explanation of the challenges faced by current water science and that relate to the need of finding analytical frameworks that contribute useful inputs to integrated management of the water resources (IWRM). As with the case of other resources, IWRM requires the analytical connection of the social and ecosystem dynamics. As a key piece within Sustainability Science the analogy of the metabolism of societies can be used to establish this connection. However, the metabolism concept needs a close examination before its joint use with other conceptions of the relations between humans and nature. After highlighting the need of considering the societal and ecosystem metabolism of socio-ecosystems as two separate but connected processes, a conceptual scheme is proposed in Chapter 2 to describe the metabolic relations between them. In Chapter 3, this scheme is adapted to the specifics of water using some of the most relevant concepts in socio- and eco-hydrology. In this way the water metabolism of socio-ecosystems is defined as the metabolism of the coupled water-human systems. Part II describes the methodological framework. In Chapter 4 the Multi-Scale Assessment of the Societal and Ecosystem Metabolism (MuSIASEM) is presented as an established framework able to deal with the scale issues and the integration of narratives. MuSIASEM is selected as a root and adapted to the analyses of coupled water-human systems. Since water presents some differences with the previous energy-focus analyses, its adaptation requires the inclusion of new scales of analysis –problemshed and watershed- and new definitions of water as a metabolite –as flow and fund. In Chapter 5 the differences and synergies between MuSIASEM and the water footprint analysis –as one of the tools of the IWRM- are highlighted. In part III four case studies are presented with two objectives. First, Chapter 6 assesses the sustainability of the metabolic patterns I Punjab and Mauritius in order to test the adaptation of MuSIASEM to water and to show how this type of analyses is made functional. Second, Chapter 7 shows how the water footprint accounting methods can complement the analysis of the water flows in MuSIASEM and how MuSIASEM, in turn an provide a space for their contextualization. Keywords: Agriculture, Complex Systems, Integrated Water Resources Management, Flow/Fund Model, Grammar, Multilevel Matrixes, MuSIASEM, Scale Issues, Socio-Ecological System, Social Metabolism, Virtual Water, Water, Water Footprint, New Water Culture.

Pharmaceuticals which are used worldwide are designed to facilitate the life for the human society and have an important role in treatment and prevention of disease for both humans and animals. They are ubiquitous in the aquatic environment and are mainly derived from municipal wastewater treatment plants (WWTPs) due to their low removal rate. Therefore, their presence in the environment is directly linked to the human impact. Various biological and abiotic processes in the environment can transform them to transformation products (TPs). In many cases, transformation is already initiated in the human body by a variety of drug-metabolizing enzymes. The metabolites formed through human metabolism present some modifications in their chemical structures that can differ in physicochemical properties to their parent compound. Once they are excreted from the human body, both the unmetabolised parent drug and their metabolites enter WWTPs by means of the sewer system. Since the WWTPs are not designed to remove completely pharmaceutical residues, the fraction not removed after the treatment will eventually end up in the receiving water bodies. Consequently, due to pharmaceutical transformations in the human body, biotransformations in WWTPs and phototransformations in surface water, they can potentially produce a high number of TPs in real world samples which makes their identification a challenge. In this thesis, two different approaches (TPs profiling and suspect screening) based on high resolution mass spectrometry (HRMS) for the detection and identification of TPs of pharmaceuticals were investigated. TPs profiling approach was applied for the identification of phototransformation products of an antiviral zanamivir (ZAN) in batch reactors filled with surface water. On the other hand, suspect screening approach was applied for evaluation of transformation, prioritization and identification of photoTPs of six iodinated contrast media in surface water. Finally, a combination of suspect and TPs profiling approach was applied for the detection of TPs of an anticonvulsant lamotrigine and its main human metabolite lamotrigine N2-glucuronide which were formed as the result of their degradation in both activated sludge and pH dependent hydrolysis. The TPs profiling approach for evaluation of these transformations is illustrated in the example of photodegradation of an antiviral ZAN with identification of its TPs in surface water (Chapter 3.). Here a set of lab- scale experiments was performed in order to determine the susceptibility of ZAN towards photodegradation under simulated and natural sunlight. The identification of the TPs was performed using hydrophilic interaction liquid chromatography coupled to high resolution mass spectrometry (HILIC-HRMS) where four photoTPs were tentatively identified and their proposed structures were rationalized by photolysis mechanisms. Kinetic experiments showed that photodegradation kinetics of ZAN in surface waters would proceed with slow kinetics since upon exposure of aqueous solutions of surface water (20 μg L-1) to simulated sunlight, ZAN was degraded with t1/2 of 3.6 h. Under natural sunlight irradiating surface water, about 30 % of the initial concentration of the antiviral disappeared within 18 days. However, when ZAN and its TPs were retrospectively screened from surface water extracts, neither the parent nor the TPs were detected. The results of this TP profiling used for the identification of TPs of ZAN, although straightforward, suggests that it is not suitable when dealing with a considerably elevated number of TPs formed in batch experiments. However, time and effort needed to be optimised for the structure elucidation of 108 photoTPs of six iodinated contrast media (ICM) (Chapter 3.). Again, the photodegradation study was performed in surface water spiked with the ICMs using a sunlight lab-scale simulator. 108 TPs were generated and each photoTP was characterised by its unique exact mass of the molecular ion and retention time and added to a suspect list. Once the suspect list was generated, the photoTPs were searched in thirteen surface water samples which were extracted using a generic solid- phase extraction method (four cartridges of different chemistries in order to retain ample number of compounds with different chemical properties). Based on their detection frequency (those TPs with the frequency higher than 50 % were deemed important), eleven TPs were prioritized and their structures elucidated by HRMS and NMR (when possible). Out of the eleven prioritised TPs, ten were formed as the result of deiodination (either by deiodination, oxidative deiodination or intramolecular elimination). In the real surface water samples, median concentration of parent compounds was 110 ngL-1 reaching up to 6 µgL-1 for iomeprol while TPs were found at median concentration of 8 ngL-1, reaching up to 0.4 µgL-1 for iomeprol TP651-B. Here detection-based prioritization served as a crucial step to reduce the number of TPs to be identified and thereby reducing costs and time for the subsequent target analysis. This time-effective approach not only guaranties that the degradation products elucidated would be found, but also that they are environmentally relevant. In summary, the proposed screening approach facilitates the evaluation of the degradation of polar compounds at a real scale with a fast detection of TPs without prior availability of the standards. Approach used for detection and identification of TPs of ICM in Chapter 3 was an example of suspect screening where the suspect list of TPs was generated at lab-scale, In Chapter 4, the work started with a suspect screening of lamotrigine (LMG) and related compounds (its human metabolites, synthetic impurities and photoTPs) which were listed from the literature and searched in wastewater and surface water samples. As the result of suspect screening, LMG, three human metabolites and a LMG synthetic impurity (OXO-LMG) were detected in the screened samples. Preliminary results showed significantly higher concentrations of OXO-LMG in wastewater effluent, suggesting its formation in the WWTPs. However, biodegradation reactors amended with mixed liquor at neutral pH showed that LMG is resistant to biodegradation with only about 5 % elimination after 6 days. Since LMG is extensively and predominantly metabolised by phase II metabolism to its N2-glucuronide, this metabolite (LMG-N2-G) was degraded following the same experimental setup. Results showed that this metabolite was the actual source of the TP detected. Additionally, in batch experiments, LMG-N2-G was transformed, following pseudo-first kinetics, to three TPs as a result of i) deconjugation (to LMG), ii) oxidation of the glucuronic acid (to LMG-N2-G-TP430) and iii) amidine hydrolysis in combination with deconjugation (to OXO-LMG). In order to further rationalize the formation of the TP OXO-LMG, the stability of LMG-N2-G and related compounds was studied as a function of pH in the range of 4 – 9. Same as during biodegradation, LMG was stable across the entire pH range tested. However, LMG-N2-G was transformed to three TPs at neutral – basic pH. They were identified as TPs formed after hydrolysis of amidine and guanidine moieties. The third TP detected was an intermediated in the guanidine hydrolysis reaction. Kinetic experiments in wastewater samples at different concentration (20 and 200 nM) and pH (pH 6.5, 7, 8, 8.5 and 9) demonstrated that while the degradation constants were concentration independent, at higher pH, LMG-N2-G degraded at higher rate. The pH-dependent stability experiments of related compounds with different nitrogen N2-substituents on the 1,2,4-triazine ring showed that reaction of amidine and guanidine hydrolysis depends on imine tautomer equilibrium whose formation depends directly on the N2-supsitutent. LMG- N2-G major abiotic TP (amidine hydrolysis TP) was detected in hospital effluent and WWTP influent samples. Having in mind the concentrations of both biotic and abiotic TPs detected, a total mass balance at two- concentration levels batch reactors was closed at 86% and 102%, respectively. In three WWTPs total mass balance of LMG-N2-G ranged from 71-102%. Finally, LMG-N2-G and its TPs were detected in surface water samples with median concentration ranges of 23–186 ngL-1. The work presented in this chapter gives a new insight into the behaviour of glucuronides of pharmaceuticals, suggesting that they might also be sources of yet undiscovered, but environmentally relevant TPs.
Els productes farmacèutics, l'ús dels quals s'estén a nivell mundial, estan dissenyats per millorar la qualitat de vida de la societat i juguen un paper clau en el tractament i la prevenció de malalties, tant en homes com en animals. Aquests compostos químics es troben de forma ubíqua en el medi ambient. Això es deu principalment a les estacions depuradores d'aigua residual (EDARs), les quals no són capaces d'eliminar de manera eficient aquest tipus de compostos, ja que no estan dissenyades amb aquesta finalitat. Per tant, la presència de fàrmacs en el medi ambient està directament relacionada amb l'activitat humana. Un cop al medi ambient, l'estructura d'aquests compostos pot ser modificada per diferents processos biològics i abiòtics, generant-se així els que es coneixen com a productes de transformació (PTs). De fet, la transformació dels fàrmacs pot iniciar-se en alguns casos en el cos humà, després de la seva administració a causa de l'activitat metabòlica dels diferents enzims que posseeix l'home. Els metabòlits formats en aquests processos presenten algunes modificacions en les seves estructures químiques pel que fa al compost original, i, en conseqüència, unes propietats fisicoquímiques diferents. Un cop excretats, tant el fàrmac original no metabolitzat com els seus metabòlits arriben a les EDARs mitjançant la xarxa de sanejament municipal d'aigües residuals. La fracció d'aquests compostos que no s'elimina en els diferents tractaments realitzats en l'EDAR, es descarrega juntament amb l'efluent de la planta als aigües receptores. El gran nombre de transformacions que poden experimentar els fàrmacs en el seu cicle de vida a causa del seu metabolisme en el cos humà, la seva biotransformació per microorganismes i la seva fototransformació per llum solar, pot generar un nombre molt elevat de PTs en el medi ambient, i, per tant, la identificació dels mateixos, necessària per avaluar el destí dels fàrmacs en el medi ambient, és un desafiament. En el desenvolupament d'aquesta tesi es van aplicar dues aproximacions analítiques diferents: a)avaluació de perfils de PTs generats en experiments a escala de laboratori i b) anàlisi qualitativa dirigida suspect screening en mostres reals, tots dues basades en espectrometria de masses d'alta resolució (HRMS) per a la detecció i identificació de PTs de productes farmacèutics. L'aproximació d'avaluació de perfils de PTs en reactors a escala de laboratori es va aplicar per identificar productes de fototransformació (fotoPTs) de l'antiviral zanamivir (ZAN) en aigua superficial. L'aproximació de suspect screening es va aplicar per prioritzar i identificar fotoPTs de sis mitjans de contrast radiològics iodats (ICM) en aigua superficial. Finalment, una combinació de les dues aproximacions es va aplicar per detectar PTs de l’anticonvulsiu lamotrigina (LMG) i del seu principal metabòlit humà, el lamotrigina-N2-glucurònid (LMG- N2-G), resultants de la seva degradació tant en fangs activats com a reaccions d'hidròlisi a diferents valors de pH.