The ATP-binding cassette (ABC) transporter, ABCA1, plays a pivotal role in reverse cholesterol transport, which is the elimination of excess sterols from peripheral cells and their transport to the liver for elimination. Early studies failed to detect significant ATPase activity, prompting the suggestion that ABCA1 was an ATP-regulated receptor, rather than an active transporter. We have provided evidence that ABCA1 can bind ATP and trap its hydrolysis product, ADP, in the presence of either ortho-vanadate or beryllium fluoride and Mg2+ or Mn2+. We have also shown that both nucleotide-binding domains (NBDs) trap nucleotide comparably, suggesting that ABCA1 is a functional ATPase. In addition, we have shown that ABCA1 can directly transport 25-hydroxycholesterol (25-OHC) in an ATP-dependent manner using a membrane vesicle uptake assay, and can do so when the physiological substrate acceptor apoA-I is replaced with BSA as a non-specific binding protein. Although more than 50 naturally occurring missense mutations and polymorphisms in ABCA1 have been identified in individuals with HDL-C levels within the lowest 5th percentile of the general population, the extent to which many of these mutations affect ABCA1 function is not known and cannot be predicted. Naturally occurring extracellular loop (ECL) mutations W590S and C1477R have both been shown to effectively eliminate the ability to mediate lipid efflux, despite the fact that the W590S mutant protein retains the ability to bind apoA-I. We show that neither mutant can transport nor efflux 25-OHC, whether in the presence of apoA-I or BSA, despite apparently full retention of the ability to bind and trap nucleotide. This suggests that these two ECL mutations inhibit transport by a mechanism that is independent of their effect on apoA-I binding. By introduction of naturally occurring mutations in the NBDs, we show that although some mutations associated with Tangier Disease, such as N935S, essentially eliminate nucleotide trapping and substrate translocation, other polymorphisms such as L1026P and T2073A associated with low HDL-C, appear to be fully functional. Lastly, we observed differences in the behaviour of both wild-type and mutant forms of ABCA1-GFP depending on whether they were expressed in insect or mammalian cell lines. / Thesis (Ph.D, Pathology & Molecular Medicine) -- Queen's University, 2010-02-12 11:14:11.381

No description available.

Les patients atteints de la maladie de Tangier présentent des niveaux très bas de lipoprotéines de haute densité (HDL), un facteur de risque pour le développement des maladies cardiovasculaires. In vivo, les HDL ont un effet protecteur important contre l’athérosclérose puisqu’elles effectuèrent le transport à rebours du cholestérol des tissus périphériques vers le foie. Or, la maladie de Tangier est causée par des mutations dans le gène du transporteur « ATP-binding cassette A1 » (ABCA1). Le modèle actuel stipule que ce transporteur assure la lipidation de l’apolipoprotéine A-I (apoA-I), la composante protéique majeure des HDL, pour former des particules HDL naissantes discoïdales. Un défaut dans la lipidation de l’apoA-I par l’ABCA1 abolit la biogénèse des HDL. Nous avons voulu étudier les sites d’interaction de l’ABCA1 avec son ligand (l’apoA-I), les voies de biogénèse impliquées, et l’implication des pré-β-HDL dans l’efflux du cholestérol par la voie de l’ABCA1. D’abord, nous avons utilisé un système de culture cellulaire (fibroblastes humaines et BHK-ABCA1-inductible) afin de déterminer les sites de liaison cellulaires de l’apoA-I, leurs localisations et l’implication de l’ABCA1. Nous avons trouvé que la majorité de l’apoA-I n’est pas associée à l’ABCA1 et, deux tiers de cet apoA-I, était à la membrane plasmique. Ensuite, Une étude plus détaillée examinait les voies de lipidation de l’apoA-I, soit au niveau de la membrane plasmique (MP), soit aux compartiments intracellulaires (CICs). Nous avons montré que la lipidation de l’apoA-I a lieu aux deux niveaux (MP et CICs) selon deux voies différentes cinétiquement. Finalement, nous avons montré que les pré-β-HDL effluent aussi (efficacement que l’apoA-I) le cholestérol par la voie de l’ABCA1. Ces observations réunies démontrent que 1) la majorité de l’apoA-I s’est trouvé non-associée à l’ABCA1; 2) deux tiers de l’apoA-I s’associent a la membrane plasmique; 3) la lipidation de l’apoA-I se fait en partie à la membrane plasmique et, par la voie de retro-endocytose du complexe apoA-I/ABCA1. / Patients affected with Tangier disease show abnormal low levels of high density lipoproteins (HDL), a risk factor for the development of cardiovascular diseases. In vivo, HDL has an important protective effect against atherosclerosis since they accomplish the reverse cholesterol transport from peripheral tissues towards the liver. However, Tangier disease is caused by mutations in the gene of the transporter “ATP-binding cassette A1” (ABCA1). The current model believe that ABCA1 promotes the lipidation of apolipoprotein AI (apoA-I), the major protein component of HDLs, to form nascent discoid HDL particles. A defect in the lipidation of apoA-I by ABCA1 abolishes the biogenesis of HDL. We wanted to study interaction sites of ABCA1 with its ligand (the apoA-I), the implicated biogenesis’s pathways, and the implication of pre-β-HDLs in the cholesterol efflux by the ABCA1 pathway. Initially, we used a cell culture system (human skin fibroblasts and BHK-ABCA1) in order to determine the cellular sites of apoA-I binding and their localizations and the implication of ABCA1. We found that the majority of apoA-I was not associated with ABCA1 and, 2 thirds of this apoA-I bound to the plasma membrane. Then, a more detailed study examined the lipidation pathways of apoA-I either at plasma membrane (PM) level, or in the intracellular compartments (ICCs). We showed that the lipidation apoA-I occurs at the two levels (PM and ICCs) with two kinetically different pathways. Finally, we showed that the pre-β-HDLs efflux (as efficiently as apoA-I) the cholesterol via the ABCA1 pathway. Taken together, these observations show that 1) the majority of apoA-I was found non-associated with ABCA1; 2) two thirds of apoA-I bind the plasma membrane; 3) the lipidation of the apoA-I occurs, in part at the plasma membrane and, in the other, by the retro-endocytosis pathway of the apoA-I/ABCA1complexe.

Doctor of Philosophy(PhD) / Atherosclerosis manifests a state of increased oxidative stress characterized by comparable lipid and protein oxidation in the affected arterial wall. While oxidative modification of low density lipoprotein (LDL) has been extensively studied, increasing attention has been focused recently on oxidation of high-density lipoproteins (HDL) and its functional consequences in relation to atherosclerosis. Oxidative modification is thought to generate “dysfunctional” HDL that has lost anti-atherosclerotic activities, including the ability to remove cholesterol from lipid-laden cells. Therefore, there has been much interest in the detection of oxidized HDL. Unfortunately, available methods to detect oxidized HDL are limited at present, in part because oxidative modification of HDL is a complex process and ‘oxidized HDL’ is not a chemically defined entity. What is known however is that conversion of methionine (Met) residues of apolipoprotein (apo) A-I to methionine sulfoxide (MetO) is a process that occurs commonly as HDL undergoes oxidative modification. For example, human apoA-I+16 (containing MetO86 or MetO112) and apoA-I+32 (MetO86 plus MetO112) are generated when apoA-I reacts with lipid hydroperoxides formed as a consequence of the lipoprotein being exposed to 1e−oxidants. The formation of MetO in apoA−I induced by 2e−oxidants (i.e., hydrogen peroxide, hypochlorous acid or myeloperoxidase/hydrogen peroxide/chloride system) is associated with an impaired ability of the apolipoprotein to facilitate reactions relevant to reverse cholesterol transport. In addition, a previous study has suggested the plasma content of apoA-I+32 to be increased in certain subjects that have an increased risk to develop cardiovascular disease (CVD). Moreover, the MetO content in circulating, HDL−associated apoA−I is elevated in type 1 diabetes, a disorder commonly associated with increased oxidative stress and a risk factor for atherosclerosis. Therefore, in the present study, an existing HPLC method was applied to HDL samples from the Fletcher−Challenge study, a nested case control study, to test the potential usefulness of MetO-containing apoA-I as a marker of oxidative stress and/or CVD in a general population. Plasma samples whose HDL contained detectable apoA-I+16 and/or apoA-I+32 had significantly elevated levels of F2-isoprostanes, a marker of in vivo lipid oxidation, consistent with MetO-containing apoA-I being a useful marker of in vivo protein oxidation. Despite this however, there was no significant difference between controls and cases in their concentrations of HDL apoA-I+16 and apoA-I+32 or F2-isoprostanes, suggesting that markers of protein and lipid oxidation are not associated with the risk of coronary heart disease (CHD) in this general population. A limitation of the Fletcher−Challenge study was that only 22% of the 534 HDL samples analyzed contained apoA-I+16 and/or apoA-I+32. In addition, the HPLC−based method used is expensive and time−consuming and may lack the sensitivity needed for apolipoproteins to clinical studies. Thus, a mouse monoclonal anti-human apoA-I+32 antibody (MOA−1) was raised using HPLC−purified apoA-I+32 as immunogen. A sensitive ELISA was then developed using a commercial anti-human apoA-I monoclonal antibody as capture and biotinylated MOA−1 as detection antibody, respectively. The assay detected lipid−free HPLC−purified human apoA-I+32 in a concentration-dependent manner and with a significantly lower limit of detection (i.e., 3 ng/mL) than the HPLC method (1 μg/mL). The ELISA also detected lipid-free apoA-I modified by 2e-oxidants (hydrogen peroxide, hypochlorous acid, peroxynitrite), and HDL oxidized by 1e- or 2e-oxidants and present in buffer or human plasma. Moreover, the extent of recognition of MetO by MOA−1 increased with increasing numbers of MetO in apoA−I, as assessed by the experiments with H2O2−oxidized forms of apoA−I mutants, in which one, two or three Met residues were replaced with Leu. Their detection was concentration-dependent, reproducible, and exhibited a linear response over a physiologically plausible range of concentrations of oxidized HDL. In contrast, MOA-I failed to recognize native apoA-I, native apoA-II, apoA-I modified by hydroxyl radicals or metal ions, or LDL modified by 2e-oxidants. Furthermore, MOA−1 did not detect other Met−containing proteins oxidized by either hypochlorous acid or hydrogen peroxide. Taken together, the results showed that recognition of oxidized proteins by MOA−1 is limited to MetO contained in apoA−I. Finally, in a pilot study, plasma samples obtained from subjects with coronary artery disease (CAD) proven by angiography, and samples from CAD patients undergoing percutaneous coronary intervention (PCI) were analyzed by the ELISA. The preliminary data obtained showed elevated levels of MetO-containing apoA-I in plasma samples of CAD patients compared to those of corresponding control subjects. Unexpectedly, levels of MetOcontaining apoA-I decreased PCI compared to before PCI. A possible explanation for these results is that HDL−associated apoA−I become displaced by acute phase proteins, such as serum amyloid A, in response to PCI. In summary, the ELISA developed here specifically detects apoA-I containing MetO in HDL and human plasma. As such it may provide a useful tool for investigating the relationship between oxidized HDL and CAD.

Doctor of Philosophy(PhD) / Atherosclerosis manifests a state of increased oxidative stress characterized by comparable lipid and protein oxidation in the affected arterial wall. While oxidative modification of low density lipoprotein (LDL) has been extensively studied, increasing attention has been focused recently on oxidation of high-density lipoproteins (HDL) and its functional consequences in relation to atherosclerosis. Oxidative modification is thought to generate “dysfunctional” HDL that has lost anti-atherosclerotic activities, including the ability to remove cholesterol from lipid-laden cells. Therefore, there has been much interest in the detection of oxidized HDL. Unfortunately, available methods to detect oxidized HDL are limited at present, in part because oxidative modification of HDL is a complex process and ‘oxidized HDL’ is not a chemically defined entity. What is known however is that conversion of methionine (Met) residues of apolipoprotein (apo) A-I to methionine sulfoxide (MetO) is a process that occurs commonly as HDL undergoes oxidative modification. For example, human apoA-I+16 (containing MetO86 or MetO112) and apoA-I+32 (MetO86 plus MetO112) are generated when apoA-I reacts with lipid hydroperoxides formed as a consequence of the lipoprotein being exposed to 1e−oxidants. The formation of MetO in apoA−I induced by 2e−oxidants (i.e., hydrogen peroxide, hypochlorous acid or myeloperoxidase/hydrogen peroxide/chloride system) is associated with an impaired ability of the apolipoprotein to facilitate reactions relevant to reverse cholesterol transport. In addition, a previous study has suggested the plasma content of apoA-I+32 to be increased in certain subjects that have an increased risk to develop cardiovascular disease (CVD). Moreover, the MetO content in circulating, HDL−associated apoA−I is elevated in type 1 diabetes, a disorder commonly associated with increased oxidative stress and a risk factor for atherosclerosis. Therefore, in the present study, an existing HPLC method was applied to HDL samples from the Fletcher−Challenge study, a nested case control study, to test the potential usefulness of MetO-containing apoA-I as a marker of oxidative stress and/or CVD in a general population. Plasma samples whose HDL contained detectable apoA-I+16 and/or apoA-I+32 had significantly elevated levels of F2-isoprostanes, a marker of in vivo lipid oxidation, consistent with MetO-containing apoA-I being a useful marker of in vivo protein oxidation. Despite this however, there was no significant difference between controls and cases in their concentrations of HDL apoA-I+16 and apoA-I+32 or F2-isoprostanes, suggesting that markers of protein and lipid oxidation are not associated with the risk of coronary heart disease (CHD) in this general population. A limitation of the Fletcher−Challenge study was that only 22% of the 534 HDL samples analyzed contained apoA-I+16 and/or apoA-I+32. In addition, the HPLC−based method used is expensive and time−consuming and may lack the sensitivity needed for apolipoproteins to clinical studies. Thus, a mouse monoclonal anti-human apoA-I+32 antibody (MOA−1) was raised using HPLC−purified apoA-I+32 as immunogen. A sensitive ELISA was then developed using a commercial anti-human apoA-I monoclonal antibody as capture and biotinylated MOA−1 as detection antibody, respectively. The assay detected lipid−free HPLC−purified human apoA-I+32 in a concentration-dependent manner and with a significantly lower limit of detection (i.e., 3 ng/mL) than the HPLC method (1 μg/mL). The ELISA also detected lipid-free apoA-I modified by 2e-oxidants (hydrogen peroxide, hypochlorous acid, peroxynitrite), and HDL oxidized by 1e- or 2e-oxidants and present in buffer or human plasma. Moreover, the extent of recognition of MetO by MOA−1 increased with increasing numbers of MetO in apoA−I, as assessed by the experiments with H2O2−oxidized forms of apoA−I mutants, in which one, two or three Met residues were replaced with Leu. Their detection was concentration-dependent, reproducible, and exhibited a linear response over a physiologically plausible range of concentrations of oxidized HDL. In contrast, MOA-I failed to recognize native apoA-I, native apoA-II, apoA-I modified by hydroxyl radicals or metal ions, or LDL modified by 2e-oxidants. Furthermore, MOA−1 did not detect other Met−containing proteins oxidized by either hypochlorous acid or hydrogen peroxide. Taken together, the results showed that recognition of oxidized proteins by MOA−1 is limited to MetO contained in apoA−I. Finally, in a pilot study, plasma samples obtained from subjects with coronary artery disease (CAD) proven by angiography, and samples from CAD patients undergoing percutaneous coronary intervention (PCI) were analyzed by the ELISA. The preliminary data obtained showed elevated levels of MetO-containing apoA-I in plasma samples of CAD patients compared to those of corresponding control subjects. Unexpectedly, levels of MetOcontaining apoA-I decreased PCI compared to before PCI. A possible explanation for these results is that HDL−associated apoA−I become displaced by acute phase proteins, such as serum amyloid A, in response to PCI. In summary, the ELISA developed here specifically detects apoA-I containing MetO in HDL and human plasma. As such it may provide a useful tool for investigating the relationship between oxidized HDL and CAD.

abstract: There are limited studies exploring the direct relationship between coconut oil and cholesterol concentrations. Research in animals and a few intervention trials suggest that coconut oil increases the good cholesterol (high density lipoprotein, HDL) and thus reduces the risk of cardiovascular disease. Preliminary research at Arizona State University (ASU) has found similar results using coconut oil as a placebo, positive changes in HDL cholesterol concentrations were observed. The goal of this randomized, double blind, parallel two arm study, was to further examine the beneficial effects of a 2g supplement of coconut oil taken each day for 8 weeks on cholesterol concentrations, specifically the total cholesterol to HDL cholesterol ratio, compared to placebo. Forty-two healthy adults between 18-40 years of age, exercising less than 150 minutes each week, non smoking, BMI between 22-35 and not taking any medications that could effect blood lipids were recruited from the ListServs at ASU. Participants were randomized to receive either a placebo capsule of flour or a coconut oil capsule (Puritan’s Pride brand, coconut oil softgels, 2g each) and instructed to take the capsules for 8 weeks. Results indicated no significant change in total cholesterol to HDL ratio between baseline and 8 weeks in the coconut oil and placebo groups (p=0.369), no significant change in HDL (p=0.648), no change in LDL (p=0.247), no change in total cholesterol (p=0.216), and no change in triglycerides (p=0.369). Blood lipid concentrations were not significantly altered by a 2g/day dosage of coconut oil over the course of 8 weeks in healthy adults, and specifically the total cholesterol to HDL ratio did not change or improve. / Dissertation/Thesis / Masters Thesis Nutrition 2017

Background Osteoarthritis (OA) is a long-term chronic disease that affects the joints and creates stiffness, pain and impaired movement. Knee osteoarthritis is the most common form of OA and affects all tissues of the joint, including bone, muscles, synovia, and cartilage. Previously, OA was accepted as only an age- or mechanical stress-related degenerative joint disease, but more recent studies suggest that OA is a heterogenous disease including inflammatory, hormonal and metabolic factors such as abdominal obesity (visceral fat), lipids (cholesterol, HDL, LDL and triglycerides) and glucose.    Aim The aim was to investigate the association of metabolic factors including fasting blood glucose, HbA1c, triglycerides, cholesterol, LDL, HDL, visceral fat, CRP and radiographic KOA in patients with symptomatic knee osteoarthritis. Methods Data were acquired from 91patients in the ages 30 – 63 experiencing symptomatic knee osteoarthritis. All subjects where divided into two groups depending on their level of fasting glucose, high versus low. Group I (n=26) had high glucose levels ≥5,6 mg/L and group II (n=65) had low glucose levels <5,6 mg/L.  Levels of HbA1c, lipids, visceral fat, CRP and radiographic KOA were then compared between the groups. Levels of fasting glucose, HbA1c and lipids (triglycerides, cholesterol, LDL, HDL) were analyzed by an accredited laboratory at the hospital of Halmstad by the department for labmedicine. CRP levels < 1 mg/L were manually analyzed with the sandwich ELISA method (enzyme-linked immunosorbent assay), which measures high-sensitive CRP (hsCRP) in serum. Visceral fat area was measured through bioelectrical impedance analysis (BIA) with InBody 770 and radiographs of the knees to obtain information about OA. Results There was a significant difference between the two groups in HbA1c, triglycerides, cholesterol and LDL p<0,05. Group I with high fasting glucose levels showed higher significant values of HbA1c, triglycerides, cholesterol and LDL than group II with low fasting glucose levels. 23% of all subjects met the requirement for metabolic syndrome according to IDF. Conclusion The findings in this study is in line with previous research and suggest that high glucose levels are associated with elevation of other metabolic factors in patients with knee osteoarthritis. However, there are several other interacting factors beyond the scope of this study, which may explain causalities. According to the findings in this study and previous research, obesity and metabolic syndrome could explain some of the connections between metabolic factors and knee osteoarthritis. Thus, further research is necessary to understand how all these metabolic factors are associated with osteoarthritis and obtain deeper knowledge about the pathogenesis and pathophysiology of the disease. / Detection and prediction of disease course in symptomatic knee osteoarthritis

La pollution des eaux constitue actuellement une préoccupation majeure aussi bien d’un point de vue sanitaire qu’environnemental. Afin d’y remédier, des procédés de dépollution efficaces, économiques et durables sont constamment développés et l’adsorption reste une méthode largement utilisée dans le traitement des eaux usées. De nombreux travaux sont consacrés à l’adsorption des polluants cationiques, mais le développement de supports adaptés pour interagir avec les contaminants anioniques est moins abordé. L’objectif principal de ce travail de thèse est de mettre en forme deux adsorbants anioniques : l’HDL (hydroxyde double lamellaire) Mg/Al et la ferrihydrite deux-lignes, et les associant à un gel d’alginate pour produire des matériaux composites pouvant être mis en oeuvre dans des réacteurs spécialisés pour l’élimination de contaminants anioniques. Les résultats de l’adsorption en réacteur batch sur les HDL et la ferrihydrite ont montré une bonne efficacité pour l’élimination de certains anions. La ferrihydrite possède une capacité plus importante que les HDL pour l’élimination des anions inorganiques. L’augmentation du rapport Mg/Al dans l’HDL a favorisé l’adsorption du méthyl orange (MO) ; un rapport Mg/Al de 3 (L3) a été séléctionné pour la suite du travail. Deux protocoles ont été évalués pour associer les solides à un gel d’alginate : l’encapsulation et la synthèse interne dans le gel. La nature des cations réticulant et la création de la macroporosité ont été étudiées. L’encapsulation du solide a diminué légèrement la capacité d’adsorption des anions, cependant, la création de la macroporosité permet de compenser cet effet du gel d’alginate. Les études sur l’adsorption du MO avec les billes encapsulées ont montré que l’augmentation de la taille des billes et de la quantité du solide L3 encapsulée a été accompagnée par une diminution de la capacité d’adsorption du MO. Le remplacement du réticulant Ca2+ par Ba2+ a permis de renforcer la structure du gel et améliorer la cinétique d’adsorption du MO, mais a entrainé une diminution de la capacité d’adsorption. La caractérisation des supports préparés par synthèse interne a montré une structure des solides différente des matériaux attendus (L3 ou ferrihydrite) avec par conséquence une réactivité différente. Des mécanismes d’élimination des anions organiques et inorganiques ont été proposés selon les différents adsorbants étudiés. Les billes ferrihydrite/alginate préparées par encapsulation ont présentés globalement une bonne capacité de rétention des anions notamment pour les anions inorganiques (phosphate, chromate et arséniate). Tandis que celles préparées par synthèse interne ne fixent que les arséniates et les phosphates par différents mécanismes essentiellement la précipitation des anions à la surface des oxyde et oxy-hydroxydes de fer. L’adsorption du MO en réacteur filtrant ouvert a montré que l’augmentation du débit d’alimentation a un effet négatif sur l’adsorption alors qu’une concentration initiale plus grande favorise l’adsorption ce qui est en concordance avec les résultats obtenus en réacteur batch. Un taux de régénération de 30% des billes ferrihydrite/alginate macroporeuse (BPENFh-Ba) a été observé en réacteur batch en utilisant une solution de chlorure d’ammonium avec la possibilité de réutiliser l’adsorbant plusieurs fois.Ce travail confirme les performances et la possibilité d’utiliser des matériaux composite d’HDL ou ferrihydrite dans un gel d’alginate pour l’élimination de composés anioniques contenus dans des eaux usées. / Water pollution is currently a major concern from both health and environmental point of view. In order to solve this problem, efficient decontamination processes, economic and sustainable are constantly being developed and adsorption process remains a widely used method in the wastewater treatment. Many studies are devoted to the adsorption of cationic pollutants, but the development of suitable supports able to interact with the anionic contaminants is less discussed. The main objective of this work is to form two anionic adsorbents: the LDH (Layered Double Hydroxide) Mg/Al and two-line ferrihydrite. These adsorbents were associated with an alginate gel in order to produce composite materials that can be implemented in continuous reactors for the removal of anionic contaminants. The adsorption studies in batch reactor on LDH and ferrihydrite confirmed good removal efficiency for some anions. Ferrihydrite showed a higher adsorption capacity than LDH for the removal of inorganic anions. Increasing the Mg/Al ratio was favorable for the methyl orange (MO) adsorption, and an Mg/Al ratio of 3 (L3) was selected for the following work. Two protocols were evaluated for the integration of adsorbent into alginate gel: encapsulation and direct synthesis in the gel structure. The effect of crosslinking cation and the making of macroporosity were studied. The encapsulation of solids decreased slightly the adsorption ability of anions; however, the macroporosity counterbalances this negative effect of alginate gel. The MO adsorption studies with encapsulated beads showed that the increase of the bead size and the amount of solid in the beads were accompanied with a decrease in the adsorption capacity. The replacement of Ca2+ by Ba2+ strengthened the gel structure and improved the adsorption kinetics but decreased the adsorption capacity. The characterization of the support material prepared by direct synthesis in alginate gel showed a different structure from the expected materials (L3 or ferrihydrite), and therefore a different reactivity. The sorption mechanisms of various organic and inorganic anions were proposed for the studied adsorbents. The ferrihydrite/alginate beads prepared by encapsulation showed a good adsorption for all the anions used in this work, especially for inorganic anions (phosphate, chromate and arsenate); while those produced by direct synthesis in alginate gel removed only arsenate and phosphate essentially by a precipitation mechanism of the anions at the surface of the iron oxides and oxy-hydroxides. The sorption of MO in a continuous reactor showed that the increasing of the flow rate had a negative effect on sorption while higher initial concentration presented a favorable effect on the sorption, which is in accordance with results obtained in batch reactor. A desorption ratio of 30% for ferrihydrite/alginate macroporous beads (BPENFh-Ba) was observed in batch reactor with the possibility of reusing the adsorbent.This study confirms the performance and the possibility to use the composite materials of LDH or ferrihydrite in an alginate gel for the removal of anionic compounds contained in wastewater.

Enzymatic and lipid transfer reactions involved in reverse cholesterol transport were studied in HDL deficient plasma systems. Fasting plasma samples were obtained from control and cholesterol fed guinea pigs as well as from a fish eye disease patient and were used to localize the enzyme LCAT among plasma lipoproteins (VLDL, LDL, and HDL). In both guinea pig and fish eye disease patient plasma, the LCAT activity was found in association with the HDL type particles. Cholesterol feeding in guinea pigs altered the properties of lipoprotein substrates for LCAT resulting in some changes, specifically: 1) decreased fractional rate of plasma cholesterol esterification and, 2) lower transfer of free cholesterol (FC) and esterified cholesterol (CE) within the lipoprotein fractions.

Les résultats des études cliniques récentes évaluant l'impact de l'augmentation du HDL-C sur la survenue des maladies cardiovasculaires n'ont pas permis de démontrer une efficacité significative de cette stratégie thérapeutique ciblée sur le HDL-C. L'évaluation de la fonctionnalité des particules HDL dans sa capacité à assurer le retour du cholestérol des tissus périphériques au foie s'avère plus importante que la simple mesure du HDL-C. L'objectif de mes travaux de recherche a été d'évaluer la fonctionnalité des particules HDL en fonction du contexte métabolique, inflammatoire et génétique de chaque individu. Mes travaux de recherche ont démontré que la niacine réduit l'athérogénicité des lipoprotéines durant la période postprandiale modulant par conséquent la fonctionnalité des particules HDL dans la voie du transport inverse de cholestérol. J'ai également montré que certaines pathologies infectieuses associées à des taux bas de HDL-C, entrainent une baisse de la capacité des particules HDL à stimuler l'efflux de cholestérol à travers le macrophage humain. En revanche, j'ai montré que des variations génétiques entrainant des taux élevés de HDL-C ne sont pas associées à une dysfonction des particules HDL dans la voie du transport inverse de cholestérol. Mes travaux soulignent l'importance de considérer que la fonctionnalité des particules HDL est modulée in vivo par des contraintes métaboliques inflammatoires et génétiques d'où il est nécessaire que les thérapies en cours de développement visant à augmenter la fonction des HDL soient résistantes aux modifications métaboliques et inflammatoires afin d'assurer une athéroprotection efficace. / HDL cholesterol as a therapeutic target has been the focus over the past 20 years mainly because its plasma concentrations are inversely correlated with the risk of coronary events. However, recent clinical trials involved the addition of niacin or CETP inhibitors to a statin in patients with CHD who had well controlled LDL-C levels have failed to show evidence of a significant reduction in cardiovascular events. Studies on the capacity of HDL to promote reverse cholesterol transport pathway have suggested that this is a better predictor of prevalent atherosclerotic disease than HDL-C itself. The main purpose of my project was to evaluate the impact of metabolic context, inflammation and genetics on modulation of HDL functionality. I have shown that extended release niacin/laropiprant treatment reduces atherogenic postprandial lipoproteins in patients with metabolic disorders stabilized by statins, having consequences on reverse cholesterol transport pathway and HDL functionality. Moreover, I have shown that infectious diseases such as HIV, induce an alteration in the capacity of HDL particles to stimulate cholesterol efflux from human macrophages. I have also shown that mutations causing high levels of HDL-C, are characterized by the presence of functional HDL in their capacity to stimulate reverse cholesterol pathway. However such patients are not systematically protected against cardiovascular disease highlighting the complex relationship between cholesterol efflux and the prediction of cardiovascular events. Those results point out the relevance of developing therapeutic approaches targeting HDL function and reverse cholesterol transport pathway.