Heat shock protein (HSP)27 is traditionally described as an intracellular chaperone and signaling molecule, but growing evidence suggests it is released from immune cells where it plays an anti-inflammatory role during atherogenesis. Previously, the O’Brien lab found that overexpression of HSP27 led to augmented HSP27 serum levels in female apolipoprotein E knockout (ApoE-/-) mice, attenuated atherogenesis, and inhibited macrophage foam cell formation via physical binding with scavenger receptor (SR)-A. However, the precise mechanism of atheroprotection remained elusive. This thesis sought to ascertain the mechanism(s) by which HSP27 prevents foam cell formation, and determine if SR-A, a key receptor involved in the uptake of lipid into macrophages, plays an important role in HSP27-mediated atheroprotection. Pre-treatment of human macrophages with recombinant HSP27 (rHSP27) inhibited acytelated low density lipoprotein (acLDL) binding and uptake independent from receptor competition effect. Reduction in uptake was associated with attenuation of expression of SR-A mRNA, total protein, and cell surface expression. To explore the signaling mechanism by which HSP27 modulated SR-A expression it was hypothesized that nuclear factor-kappa B (NF-kB), a major regulator of many atherosclerosis gene programs, is altered by extracellular HSP27. Indeed, rHSP27 markedly activated NF-kB signaling in macrophages. Using an inhibitor of NF-kBsignaling there was an attenuation of rHSP27-induced inhibition of SR-A gene and protein expression, as well as lipid uptake, suggesting that SR-A expression is regulated by NF-kB activation. Lastly, to investigate if SR-A is required for HSP27-mediated atheroprotection in vivo, ApoE-/- and ApoE-/-SR-A-/- mice fed a high fat diet were treated with rHSP25, the mouse orthologue of HSP27, or PBS for 3 weeks. While rHSP25 therapy equally reduced serum cholesterol levels in the mouse cohorts, aortic atherogenesis, assessed using en face and sinus cross-sectional analyses, was attenuated in ApoE-/- mice but not ApoE-/-SR-A-/- mice. In conclusion, rHSP27 inhibits foam cell formation by downregulating SR-A expression. This effect may be associated with NF-kB activation. Reductions in atherosclerotic burden by rHSP27 require SR-A, and are independent of changes in serum cholesterol levels, highlighting the importance of macrophage lipid uptake in atherogenesis. Results presented in this thesis demonstrate that SR-A is a major target for HSP27 atheroprotection in the vessel wall, and provide an impetus for further studies that investigate the potential therapeutic value of HSP27.

Sepsis claims over 215,000 lives in the US annually. Inducible glucocorticoids (iGC) is produced during sepsis. However, the precise effects of iGC in sepsis remain unclear due to a lack of appropriate animal models. Glucocorticoid (GC) insufficiency is associated with a marked increase in mortality and occurs in 60% of severe septic patients. Yet the conclusion of GC therapy on septic patients is still controversial. Scavenger receptor class B type I (SR-BI) in the adrenal mediates the selective uptake of cholesteryl ester from lipoproteins for GC synthesis. SR-BI-/- mice completely lack iGC during sepsis and are highly susceptible to septic death, which presents SR-BI-/- mice as a GC insufficient model. However, SR-BI-/- mice display multiple defects contributing to septic death, making it difficult to study iGC by using these mice. Therefore, we utilized adrenal-specific SR-BI-/- mice (ADR-T SR-BI-/-) generated by adrenal transplantation. As expected, the ADR-T SR-BI-/- mice failed to generate iGC under cecal ligation and puncture (CLP)-induced sepsis and showed a significantly higher mortality than the control mice, demonstrating that iGC is essential for preventing septic death. High blood urea nitrogen (BUN) was observed in the ADR-T SR-BI-/- mice but not in the control mice in CLP, indicating that iGC protects kidney injury in sepsis. Plasma IL-6 was remarkably higher in the ADR-T SR-BI-/- mice than the control mice, demonstrating an anti-inflammatory effect of iGC in sepsis. The ADR-T SR-BI-/- mice also displayed significantly lower phagocytic activity of monocytes and neutrophils in the blood and lower activation of T cells in the spleen compared to the control mice in CLP, suggesting that iGC is immunomodulatory in sepsis. Low-dose GC supplementation significantly improved the survival of SR-BI-/- mice in CLP, but did not increase the survival rate of SR-BI+/+ mice in CLP, indicating that GC supplementation improves the survival specifically in mice with adrenal insufficiency. Overall, we revealed that iGC is essential for sepsis survival. iGC prevents kidney damage, modulates inflammatory responses and exerts immunomodulatory functions in sepsis. GC supplementation specifically improves survival of individuals with adrenal insufficiency in sepsis.

Heat shock protein (HSP)27 is traditionally described as an intracellular chaperone and signaling molecule, but growing evidence suggests it is released from immune cells where it plays an anti-inflammatory role during atherogenesis. Previously, the O’Brien lab found that overexpression of HSP27 led to augmented HSP27 serum levels in female apolipoprotein E knockout (ApoE-/-) mice, attenuated atherogenesis, and inhibited macrophage foam cell formation via physical binding with scavenger receptor (SR)-A. However, the precise mechanism of atheroprotection remained elusive. This thesis sought to ascertain the mechanism(s) by which HSP27 prevents foam cell formation, and determine if SR-A, a key receptor involved in the uptake of lipid into macrophages, plays an important role in HSP27-mediated atheroprotection. Pre-treatment of human macrophages with recombinant HSP27 (rHSP27) inhibited acytelated low density lipoprotein (acLDL) binding and uptake independent from receptor competition effect. Reduction in uptake was associated with attenuation of expression of SR-A mRNA, total protein, and cell surface expression. To explore the signaling mechanism by which HSP27 modulated SR-A expression it was hypothesized that nuclear factor-kappa B (NF-kB), a major regulator of many atherosclerosis gene programs, is altered by extracellular HSP27. Indeed, rHSP27 markedly activated NF-kB signaling in macrophages. Using an inhibitor of NF-kBsignaling there was an attenuation of rHSP27-induced inhibition of SR-A gene and protein expression, as well as lipid uptake, suggesting that SR-A expression is regulated by NF-kB activation. Lastly, to investigate if SR-A is required for HSP27-mediated atheroprotection in vivo, ApoE-/- and ApoE-/-SR-A-/- mice fed a high fat diet were treated with rHSP25, the mouse orthologue of HSP27, or PBS for 3 weeks. While rHSP25 therapy equally reduced serum cholesterol levels in the mouse cohorts, aortic atherogenesis, assessed using en face and sinus cross-sectional analyses, was attenuated in ApoE-/- mice but not ApoE-/-SR-A-/- mice. In conclusion, rHSP27 inhibits foam cell formation by downregulating SR-A expression. This effect may be associated with NF-kB activation. Reductions in atherosclerotic burden by rHSP27 require SR-A, and are independent of changes in serum cholesterol levels, highlighting the importance of macrophage lipid uptake in atherogenesis. Results presented in this thesis demonstrate that SR-A is a major target for HSP27 atheroprotection in the vessel wall, and provide an impetus for further studies that investigate the potential therapeutic value of HSP27.

Les plastiques sont devenus les matériaux les plus utilisés pour la préservation de denrées alimentaires. Malgré leurs nombreux avantages (transparence, prix, modularité, propriété barrière aux gaz), leur imperméabilité à l’oxygène doit être sans cesse améliorée afin d’éviter aux denrées de subir diverses réactions d’oxydation et ainsi prolonger leurs durées de conservation. Pour cela, une solution envisageable consiste à combiner à des polymères, des catalyseurs métalliques pour piéger l’oxygène. Cette possibilité a été étudiée dans cette thèse. Après un premier chapitre introductif, le deuxième décrit les propriétés physico-chimiques des différents catalyseurs métalliques sélectionnés (stabilité, constantes de protonation, constante de complexation envers le cobalt(II) et le manganèse (II), capacité d’oxygénation, électrochimie). Le troisième chapitre est consacré à leur utilisation en tant qu’oxydant métallique de substrats organiques. Enfin le quatrième et dernier chapitre présente une étude à l’état solide de l’oxydation de polymères en présence de catalyseurs métalliques. Une large part de ce dernier chapitre a été consacrée à la caractérisation des produits d’oxydation. / Plastics are becoming the most used material for food and drinks preservation. Despite their numerous advantages (transparency, price, modularity, gas barrier property), their oxygen barrier properties have to be improved to avoid hazardous oxidation reactions to foods and drinks and so to increase their shelf-life. One alternative to solve this problem is to combine polymers and metal catalysts to scavenge oxygen. This possible strategy was investigated during this Ph.D. project. After a first introductive chapter, the second depicts the physico-chemical properties of selected metal catalysts (stability, protonation constants, complexation constants towards cobalt(II) and manganese(II), oxygenation capacity, electrochemistry). The third chapter is devoted to their use as powerful oxidant of organic substrate. Finally, the last chapter is dedicated to a solid-state study of polymer oxidation in the presence of metal catalysts. A large part of this last chapter has been devoted to the thorough elucidation of the nature/structure of the oxidized product(s).

A Economia Solidária (ES) é uma forma alternativa de pensar a atividade econômica em que valores como equidade, solidariedade e cooperação estão no centro das relações dos agentes. Os empreendimentos econômico-solidários trazem em seu cerne os valores preconizados pela ES e embrenham no desafio de construir uma forma diferenciada de produção e distribuição dos resultados do trabalho. Contudo, atuam em um mercado guiado pela racionalidade concorrencial e nesse contexto surge a indagação sobre como alinhar as necessidades de competitividade demandadas pelo mercado, com os princípios da organização econômico-solidária. Assim, o objetivo do trabalho é analisar a organização e a gestão de empreendimentos autogestionários, destacando as práticas distintivas, comparativamente aos modelos teóricos de organização e gestão, que permitem conciliar as demandas concorrenciais com os princípios solidários. Aplicando a metodologia de observação participante em três cooperativas de catadores de materiais recicláveis, o trabalho destaca, entre outras, três práticas distintivas centrais: a configuração coletiva do processo de tomada de decisão e gestão, que requer uma série de outras características, tais como, acesso à informação e conhecimento de causa; os critérios utilizados na tomada de decisão consideram os diversos objetivos dos interessados além do objetivo de desempenho econômico; e há a possibilidade de negociação sobre a produtividade do trabalho. Apresenta-se detalhadamente a organização das três cooperativas estudadas, descrevendo a configuração produtiva, a estrutura do processo decisório, o controle operacional e a distribuição das sobras. / Solidarity Economy (ES) is an alternative way of thinking of the economic activity where values as fairness, solidarity and cooperation are at the center of agents relation. The Social Enterprises are economic ventures that brings in their heart the principles of the ES and engage on the challenge of construct a differently way of production, considering new ways of work relation and organization, and results distribution. However, this ventures operate on a competitive market, where the predominant rationality is the competition and not the cooperation or solidarity. On this context the main goal of this research is to analyze the production organization and the management process of the self-management enterprises and highlight the distinctive aspects on management and/or work organization, compared to the theoretical models. Participant observation methodology was applied within three Scavengers Cooperatives (sorting of recycling materials). The paper highlights, among others, three central distinct practices: the collective configuration of the decision-making process and management that requires a number of other characteristics, such as, free information access and knowingly of the facts of the enterprise; the criteria used in the decision making that consider several objectives of stakeholders beyond the scope of economic performance; and the possibility of negotiate the work productivity. The research presents the organization of the three cooperatives studied in details, describing the production configuration, the structure of the decision making process and the operational control and distribution of surplus.

Orientador: Adriana Zerlotti Mercadante / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-11T02:48:48Z (GMT). No. of bitstreams: 1 Barreto_GiselaPizarrodeMattos_D.pdf: 1819752 bytes, checksum: 73ae705ff5b48a9b194ef27159fe6ab1 (MD5) Previous issue date: 2008 / Resumo: O aumento da demanda dos consumidores por maior qualidade de vida tem impulsionado o interesse da indústria de alimentos pelo segmento de alimentos funcionais, definidos como qualquer alimento com potencial para conferir benefícios à saúde do consumidor. Os compostos bioativos, como os carotenóides, compostos fenólicos e ácido ascórbico, presentes em alimentos estão envolvidos nestas ações benéficas. Inicialmente, foi realizado um estudo com alguns padrões de carotenóides, que não somente apresentam atividade anti-radical livre como também são responsáveis pela cor de vários alimentos. Foi avaliado o comportamento destes carotenóides, em diferentes concentrações, em relação à sua atividade anti-radical livre (ABTS) e parâmetros de cor CIELAB. Os resultados revelaram que, o número de ligações duplas conjugadas e a presença do grupo carbonila influenciaram tanto na atividade anti-radical livre como nos parâmetros de cor dos carotenóides, assim como a concentração. Posteriormente, para a determinação da atividade anti-radical livre de extratos complexos, pelo método ABTS, foram preparadas no laboratório extratos de polpas de 18 frutas tropicais, aplicando análises estatísticas multivariadas, especificamente Análise de Componentes Principais (ACP) e Análise Hierárquica de Agrupamentos (AHA), para caracterizar e agrupar as 18 polpas de frutas e verificar a correlação entre atividade anti-radical livre e alguns constituintes químicos que apresentam funcionalidade como ácido ascórbico, compostos fenólicos, flavonóides e carotenóides totais. As frutas foram agrupadas em 5 grupos, segundo análise hierárquica de agrupamentos (AHA). Na Análise de Componentes Principais, observou-se que a atividade antioxidante apresentou alta correlação com os fenóis (r = 0,99) e flavonóides (r = 0,86) totais, porém apresentou baixa correlação com ácido ascórbico (r = 0,02) e carotenóides totais (r= 0,08). Em especial, foi estudado o caju, pois, o processamento de suco a partir do pseudofruto (pedúnculo) de caju gera como subproduto um bagaço, que constitui uma rica fonte de compostos fenólicos, ácido ascórbico (AA) e carotenóides. Considerando que este bagaço pode ser transformado para produzir ingredientes funcionais, o caju clone CCP-076, proveniente da Estação Experimental de Paraipaba (CE) da Embrapa Agroindústria Tropical, foi prensado e o bagaço obtido foi umidificado e novamente prensado até exaustão (retentado), sendo a fase líquida filtrada (extrato bruto) e concentrada (extrato concentrado). O retentado apresentou maior teor de fenóis totais, de carotenóides totais e de atividade anti-radical livre, além de apresentar cor amarela mais intensa. Por outro lado, o extrato concentrado apresentou o maior teor de AA. Os principais carotenóides encontrados, tanto no retentado como nos extratos, foram all-trans-b- criptoxantina e all-trans-b-caroteno. Uma outra fruta também analisada em separado foi o mamão papaia (Carica papaya cv. Golden) que tem uma grande demanda de exportação para os mercados da América do Norte e Europa; entretanto, é uma fruta climatérica sendo susceptível a perdas após a colheita. O mamão-papaia foi dividido em três grupos: controle (sem tratamento), etileno (tratamento com etileno) e 1- metilciclopropeno (1-MCP) (tratamento com 1-MCP). A composição de carotenóides e os parâmetros de cor de todos os grupos foram determinados durante o amadurecimento da fruta. A presença do gás etileno e 1-MCP reduziram o teor de carotenóides e favoreceram a biossíntese de carotenóides acíclicos. Entretanto, mesmo com menor teor de carotenóides que o grupo controle, os valores de cor (a*, b*) foram similares nos três grupos durante o amadurecimento / Abstract: The increased demand for high life quality has been associated to healthy food, increasing the industry interest for functional foods, which are defined as any food or bioactive compounds with potential to be benefic for the consumer health. The bioactive compounds found in foods, such as carotenoids, phenolic compounds and ascorbic acid (AA) are involved with these benefic actions. Firstly, some carotenoid standards in different concentrations were evaluated regarding not only their radical scavenger activity, but also their color parameters (CIELAB), since these pigments are responsible for the color of many foods. The results showed that the number of conjugated double bonds, presence of carbonyl group and concentration influenced both the radical scavenger activity and the color parameters. After that, the radical scavenger activity of complex extracts were determined, by ABTS assay, obtained from pulps extracts prepared in the laboratory from 18 tropical fruit applying Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) to characterize and grouped the 18 extracts, as well as to correlate radical scavenger and the bioactive compounds, such as AA, phenolic compounds, flavonoids and total carotenoids. The fruits were divided into five groups, following Hierarchical Cluster Analysis (HCA). Principal Component Analysis (PCA) indicated that antioxidant activity showed high correlation to total phenolic compounds (r= 0.99) and flavonoids (r= 0.86); however, correlation was found to be very poor with AA (r= 0.02) and total carotenoid levels (r= 0.08). The cashew-apple was specially evaluated since the processing of juice from the cashew-apple fruit generates a pomace that constitutes a rich source of phenolic compounds, AA and carotenoids. Considering that this pomace can be processed into functional ingredients, the cashew-apple clone CCP-076, from Estação Experimental de Paraipaba (CE) at Embrapa Agroindústria Tropical, was pressed and than the pomace formed was humidified and pressed again until exhaustion (retentado), followed by filtration (raw extract) and concentration (concentrated extract). The retentado presented significantly higher total phenol and total carotenoid contents, higher free radical scavenger activity, as well as higher yellow color intensity, indicated by higher b* and chroma (C*) values. On the other hand, the concentrated extract showed the highest AA content. The major carotenoids identified in both retentado and extracts were all-trans-b-cryptoxanthin and all-trans-b-carotene. These results showed that the retentado has a good potential to be applied as a functional ingredient. Another fruit also specially analyzed was the papaya fruit (Carica papaya cv. Golden) that has a great export demand to North American and Europe markets; however, since it is a climacteric fruit, it is susceptible to postharvest losses. The papaya fruits were divided into three groups: control (without treatment), ethylene (treated with ethylene) and 1-methylcyclopropene (1-MCP) (treated with 1-MCP). In all groups, carotenoid composition, carried out by HPLC, and color parameters were evaluated during ripening. The treatment with ethylene and 1-MCP decreased carotenoid accumulation and enhanced acyclic carotenoid biosynthesis. Although the gas treated fruits presented lower carotenoid content as compared to the control, the color values (a*, b*) were similar in treated and non-treated fruits during ripening / Doutorado / Doutor em Ciência de Alimentos

The deep-sea is the largest habitat on earth, containing over 90 percent of the world’s oceans and home to over 20,000 species. Deep-sea ecosystems are increasingly impacted by human activities including fishing and oil extraction. To understand potential impacts on deep-sea food webs, it is crucial to gather baseline data in these systems. I quantified the trophic interactions of three groups of deep-water animals across a range of trophic levels living in the northern and eastern Gulf of Mexico using stable isotope analysis. First, I propose methods for correcting δ15N values for the presence of nitrogenous metabolic waste products (e.g., urea) in muscle tissue using chemical extractions and/or species-specific mathematical normalizations. Significant differences in δ15N, %N, and C:N values as a result of extractions were observed in eight of ten shark and all three hagfish species. The δ15N values increased, but shifts in %N and C:N values were not unidirectional. Mathematical normalizations for δ15N values were successfully created for four shark and two hagfish species. I then describe the trophic interactions of three consumer assemblages. Carbon isotopic values indicate a heavy reliance on allochthonous nutrient inputs from surface waters. Nitrogen isotopic values reveal somewhat atypical taxa as top predators in the deep sea. Shark, teleost, and invertebrate species across a wide range of body sizes are feeding at a similar trophic level. This apparent lack of size structuring could be the result of a high degree of opportunistic scavenging or perhaps feeding at many trophic levels simultaneously in an oligotrophic system. There was a high degree of isotopic niche overlap among species within each consumer assemblage, perhaps the result of limited nutrient resources in the deep-sea. In general, individuals from the northern sampling stations displayed higher δ13C and δ15N values than those from the eastern sites. With the exception of a few species, there were no strong relationships between body size and isotopic values. The present study is among the first characterizations of the trophic structure of deep-sea organisms in the Gulf of Mexico and establishes system baselines for future studies describing deep-water systems and investigating anthropogenic impacts.

Heat shock protein (HSP)27 is traditionally described as an intracellular chaperone and signaling molecule, but growing evidence suggests it is released from immune cells where it plays an anti-inflammatory role during atherogenesis. Previously, the O’Brien lab found that overexpression of HSP27 led to augmented HSP27 serum levels in female apolipoprotein E knockout (ApoE-/-) mice, attenuated atherogenesis, and inhibited macrophage foam cell formation via physical binding with scavenger receptor (SR)-A. However, the precise mechanism of atheroprotection remained elusive. This thesis sought to ascertain the mechanism(s) by which HSP27 prevents foam cell formation, and determine if SR-A, a key receptor involved in the uptake of lipid into macrophages, plays an important role in HSP27-mediated atheroprotection. Pre-treatment of human macrophages with recombinant HSP27 (rHSP27) inhibited acytelated low density lipoprotein (acLDL) binding and uptake independent from receptor competition effect. Reduction in uptake was associated with attenuation of expression of SR-A mRNA, total protein, and cell surface expression. To explore the signaling mechanism by which HSP27 modulated SR-A expression it was hypothesized that nuclear factor-kappa B (NF-kB), a major regulator of many atherosclerosis gene programs, is altered by extracellular HSP27. Indeed, rHSP27 markedly activated NF-kB signaling in macrophages. Using an inhibitor of NF-kBsignaling there was an attenuation of rHSP27-induced inhibition of SR-A gene and protein expression, as well as lipid uptake, suggesting that SR-A expression is regulated by NF-kB activation. Lastly, to investigate if SR-A is required for HSP27-mediated atheroprotection in vivo, ApoE-/- and ApoE-/-SR-A-/- mice fed a high fat diet were treated with rHSP25, the mouse orthologue of HSP27, or PBS for 3 weeks. While rHSP25 therapy equally reduced serum cholesterol levels in the mouse cohorts, aortic atherogenesis, assessed using en face and sinus cross-sectional analyses, was attenuated in ApoE-/- mice but not ApoE-/-SR-A-/- mice. In conclusion, rHSP27 inhibits foam cell formation by downregulating SR-A expression. This effect may be associated with NF-kB activation. Reductions in atherosclerotic burden by rHSP27 require SR-A, and are independent of changes in serum cholesterol levels, highlighting the importance of macrophage lipid uptake in atherogenesis. Results presented in this thesis demonstrate that SR-A is a major target for HSP27 atheroprotection in the vessel wall, and provide an impetus for further studies that investigate the potential therapeutic value of HSP27.

No description available.

<p>Class A scavenger receptors are multifunctional transmembrane glycoproteins that mediate macrophage functions like phagocytosis and endocytosis. The macrophage receptor with collagenous structure (MARCO) is one such receptor. It has been shown that the extracellular cysteine-rich domain of MARCO is responsible for ligand binding, but the role of the cytoplasmic domain in ligand uptake is unclear. The aim of the studies presented in this thesis is to characterize the role of the cytoplasmic domain of MARCO and to characterize the molecular pathway of MARCO-mediated endocytosis.</p> <p>Full-length human MARCO (hMARCO) and Δ1-34hMARCO, which lacks the first thirty-four amino acids were created in order to determine whether amino acids 1-34 contained residues required for receptor internalization and surface expression. The constructs were stably expressed in HEK293T cells and found to have similar levels of surface expression and same rate of internalization without ligand. Interestingly, hMARCO, but not Δ1-34hMARCO, surface expression was up-regulated upon ligand incubation.</p> <p>In order to ascertain the importance of clathrin, dynamin and actin in MARCO-mediated endocytosis, specific endocytic inhibitors were used. MARCO-mediated ligand uptake was inhibited when clathrin and actin polymerization and, dynamin functions were impaired by these inhibitors. Furthermore, ligand uptake by Δ1-34hMARCO-expressing HEK293T was insensitive to inhibitors of clthrin and dynamin but not inhibitors of actin.</p> <p>In conclusion, MARCO mediates endocytosis via a clathrin-mediated, dynamin-dependent pathway that involves actin. Amino acids 1-34, are required clathrin and dynamin but not actin functions during MARCO-mediated endocytosis. Additionally, amino acids 1-34 might also be important for MARCO recycling but not receptor internalization or surface expression.</p> / Master of Science (MSc)