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Eicosapentaenoic acid (EPA) is a polyunsaturated fatty acid (PUFA) with neuroprotective and cardioprotective properties. It is thought that some of the actions of EPA may be attributed to its elongated metabolite, the PUFA docosapentaenoic acid (DPA). Docosahexaenoic acid (DHA) and arachidonic acid (AA) are bioactive PUFA ubiquitously expressed in neural tissues. EPA and AA can be converted by cyclooxygenase (COX) to prostanoids and by lipoxygenase (LOX) to hydroxy fatty acids. PUFA can also be converted to ethanolamides in the brain. These mediators are involved in physiological and pathological processes in many bodily systems. The purpose of this study was to examine the production of eicosanoids, hydroxy fatty acids and fatty acid ethanolamides in young and aged rat brain following EPA or DPA enriched diets. The effects of specific PUFA on human platelet eicosanoid production were also investigated as these mediators play a role in adhesion and aggregation. Liquid chromatography coupled to tandem mass spectrometry (LC/ESI-MS/MS) assays were developed and used to measure lipid mediators in rat brain and human platelets. Ageing in rat brain was accompanied with several changes in the prostanoid and hydroxy fatty acid profiles. Supplementing the diet with EPA or DPA at a daily dose of 200 mg/kg for 8 weeks prevented these changes and decreased levels of PGE2. DPA changed the profile of hydroxy fatty acids synthesised in the brain tissue of young animals. This study has shown that levels of eicosapentaenoylethanolamide (EPA-EA) increase in the brain as a result of ageing and that this is accompanied by an increase in levels of anandamide. Feeding aged animals EPA or DPA further increased the levels of EPA-EA but prevented any change in the level of anandamide. Niacin is used to treat hypercholesterolaemia although it is associated with an unpleasant PGD2 mediated skin flush. This exploratory study has shown that human platelets treated with niacin did not show any changes in their prostanoid and hydroxy fatty acid profiles. Platelets treated with EPA showed increased production of TXB3 and 12-HEPE. Niacin augmented the effects of EPA on human platelet mediator synthesis. Overall, this study has demonstrated that EPA can change brain and platelet lipid mediator synthesis and has provided evidence that could explain some of the neuroprotective and cardioprotective actions of this PUFA.

Tesis por compendio / [ES] La presente tesis doctoral presentaba como principal objetivo contribuir al conocimiento sobre estrategias de alimentación y técnicas de producción que puedan mejorar la sostenibilidad del sector acuícola a través de un enfoque multidisciplinar, evaluando el crecimiento, la salud y la calidad sensorial de los peces, así como la producción de especies vegetales. También se evaluó El impacto ambiental de las diferentes propuestas mediante el análisis del Ciclo de Vida (LCA) como parte de la discusión general de la tesis. El primer estudio evaluó el efecto de la sustitución de diferentes niveles de aceite de pescado (FO) por aceites vegetales en piensos para la seriola mediterránea (Seriola dumerili, Risso, 1810) en el perfil de ácidos grasos (FA) de diferentes tejidos. El perfíl de FA de la grasa visceral, hígado, músculo rojo y blanco cambiaron cuando se modificó la fuente lipídica en el pienso, mientras que el perfíl de FA del tejido cerebro fue bastante resistente a su alteración por los cambios en la dieta. Estos nuevos datos podrían ayudar a evaluar si las funciones fisiológicas claves se conservan cuando los peces son alimentan con dietas con niveles bajos de FO, así como para definir las necesidades de FA en la dieta de la S. dumerili para mejorar la sostenibilidad de la producción y el bienestar de los peces. El segundo estudio evaluó los efectos de un periodo de realimentación en la composición de ácidos grasos en el músculo de la seriola mediterránea. Después de 109 días durante los cuales los peces fueron alimentados con un pienso a base de aceite de pescado (FO 100) o un pienso (FO 0) en la que el FO fue sustituido completamente por aceites vegetales, todos los peces alimentados con la dieta FO 0 fueron sometidos a una realimentamentación con un pienso FO 100, durante 90 días. Un recebo de 90 días puede mejorar parcialmente el perfil de FA en los músculos de la seriola mediterránea previamente alimentada con dietas a base de aceite vegetal.El tercero estudio analizó los efectos de dos densidades de producción (baja - ALD, 3,81 kg m¿3 frente a alta - AHD, 7,26 kg m¿3) sobre el crecimiento, la salud y la calidad de la carne de la trucha arco iris (Oncorhynchus mykiss, Walbaum 1792) así como el rendimiento y calidad microbiológica de la lechuga (Lactuca sativa, L.) producida en un sistema acuapónico de baja tecnología en comparación con el cultivo hidropónico (HYP). Las producciones de trucha arco iris y lechuga fueron exitosas en el sistema acuapónico probado, mientras que la densidad de población no afectó el crecimiento de los peces ni a la calidad de la carne. En el cuarto estudio se evaluó el efecto de la sustitución dietaria de la harina de pescado (FM) por harina de Hermetia illucens (Linnaeus, 1758), parcialmente desgrasada (HI) sobre el crecimiento, la digestibilidad del alimento, la morfología intestinal y la calidad del filete de la trucha arco iris producida en un sistema acuapónico de baja tecnología. El crecimiento de los peces y la calidad del filete no se vieron esencialmente afectados por la sustitución de harina de pescado al 25% con HI. Sin embargo, con una sustitución del 50%, se detectó ciertos efectos sobre la histología intestinal y el color del filete y las características nutricionales, que justifican una mayor investigación. en base a los resultados de esta tesis, los aceites vegetales y la harina de insecto son ingredientes alternativos prometedores para ser incluidos en las dietas de especies marinas y de agua dulce de alto valor económico. Sin embargo, las formulaciones futuras deberán centrarse no solo en las características nutricionales y el coste de los ingredientes, sino también en su impacto ambiental. La acuaponía es una técnica prometedora para la cría de especies de agua dulce de alto valor como la trucha arco iris. Además, la optimización de la densidad de producción de peces en acuaponía es un aspecto clave para obtener altos rendimientos y una buena calidad del producto. / [CA] La present tesi doctoral va tenir com a objectiu contribuir al coneixement sobre estratègies d'alimentació i tècniques de producció que puguin millorar la sostenibilitat del sector aqüícola a través d'un enfocament multidisciplinari, avaluant el creixement, la salut i la qualitat sensorial dels peixos, així com la producció de espècies vegetals. El impacte ambiental de les diferents propostes també es va avaluar mitjançant l'anàlisi de l'Cicle de Vida (LCA) com a part de la discussió general de la tesi. En el primer estudi es va avaluar la substitució de diferents nivells d'oli de peix per olis vegetals en pinsos per a la seriola Mediterrània (Seriola dumerili, Risso, 1810) en el perfil d'àcids grassos (FA) de diferents teixits. El segon estudi es van avaluar els efectes d'un període de realimentació en la composició d'àcids grassos en el múscul de la seriola mediterrània. Un rentat de 90 dies pot millorar parcialment el perfil de FA en els músculs de la seriola mediterrània prèviament alimentada amb dietes a base d'oli vegetal. El tercer estudi va estudiar els efectes de dues densitats de producció (baixa - ALD, 3,81 kg m-3 enfront de alta - AHD, 7,26 kg m-3) sobre el creixement, la salut i la qualitat de la carn de la truita (Oncorhynchus mykiss, Walbaum 1792) i el rendiment i qualitat microbiològica de l'enciam (Lactuca sativa, L.) produïda en un sistema aquapònic de baixa tecnologia en comparació amb el cultiu hidropònic (HYP). En conclusió, les produccions de truita i enciam van tindre èxit en el sistema aquapònic provat, mentre que la densitat de població no va afectar al creixement dels peixos ni a la qualitat de la carn. En el quart estudi es va avaluar l'efecte de la substitució dietària de la farina de peix amb farina de Hermetia illucens (Linnaeus, 1758), parcialment desgreixada (HI) sobre el creixement, la digestibilitat de l'aliment, la morfologia intestinal i la qualitat del filet de la truita produïda en un sistema aquapònic de baixa tecnologia, el creixement dels peixos i la qualitat de l'filet no es van veure essencialment afectats per la substitució de farina de peix al 25% amb HI, mentre que amb una substitució del 50%, es va detectar certs efectes sobre la histologia intestinal i el color del filet i les característiques nutricionals, que justifiquen una major investigació. En conclusió, en base als resultats d'aquesta tesi, els olis vegetals i la farina d'insecte són ingredients alternatius prometedors per a ser inclosos en les dietes d'espècies marines i d'aigua dolça d'alt valor econòmic. No obstant això, les formulacions futures hauran de centrar no només en les característiques nutricionals i el cost dels ingredients, sinó també en el seu impacte ambiental. La aquaponia és una tècnica prometedora per a la cria d'espècies d'aigua dolça d'alt valor com la truita. A més, l'optimització de la densitat de producció de peixos en aquaponia és un aspecte clau per obtenir alts rendiments i una bona qualitat del producte, al temps que es redueix l'impacte ambiental, assegurant la salut i el benestar dels peixos. / [EN] The present PhD thesis aimed at improving the knowledge on feeding strategies and rearing techniques that might improve the sustainability of the aquaculture sector through a multidisciplinary approach, evaluating fish growth performance, health and quality, and vegetable production. The environmental impact of the different solutions was also assessed using a Life Cycle Assessment (LCA) analysis as a part of thesis general discussion. The first contribution evaluated how replacing different levels of fish oil by vegetable oils in the diet of Mediterranean yellowtail, Seriola dumerili (Risso, 1810), affects the fatty acids (FA) signature, i.e. overall FA profile, in different tissues. The FA signatures of red and white muscle, liver, and visceral fat tissues changed when the dietary FA source changed, whereas FA signatures in the brain were rather robust to such dietary changes. The second contribution assessed the effects of a wash-out on the fatty acid composition in the muscles of Mediterranean yellowtail. A 90-d wash-out can partially improve the FA profile in muscles of Mediterranean yellowtail previously fed vegetable oil-based diets. The third contribution investigated the effects of two stocking densities (low - ALD, 3.81 kg m¿3 vs. high - AHD, 7.26 kg m¿3) on the growth, health, and flesh quality of rainbow trout (Oncorhynchus mykiss, Walbaum 1792) and the yield and microbiological quality of lettuce (Lactuca sativa, L.) produced in a low-tech aquaponic system compared to hydroponic cultivation (HYP). Rainbow trout and lettuce productions were successful in the tested aquaponic system, whereas stocking density did not affect fish growth or flesh quality. The fourth contribution evaluated the effects of dietary substitution of fishmeal with partially defatted (Hermetia illucens, Linnaeus, 1758,) meal (HI) on the growth, feed digestibility, gut morphology, and fillet quality of rainbow trout reared in a low-tech aquaponic system fish growth and fillet quality were essentially unaffected by a 25% fish meal replacement with HI, whereas at a replacement rate of 50%, certain effects on gut histology and fillet colour and nutritional characteristics were detected, which warrant further investigation. In conclusion, based on the results of the present thesis, vegetable oils and insect meal are promising alternative ingredients to be included in diets for high-value marine and freshwater carnivorous species. Nevertheless, future aquafeed formulations will be required to focus not only on the nutritional characteristics and costs of the ingredients but also on their environmental impact. Aquaponics is a promising technique to be used for rearing high-value freshwater species such as rainbow trout. Furthermore, the optimization of fish stocking density in aquaponics is a key aspect to obtain high product yields and quality, reducing the environmental impact as well as assuring fish health and welfare. / Thesis written with the financial contribution of ECCEAQUA project (MIUR; CUP: C26C18000030004) / Bordignon, F. (2022). Feeding Strategies and Rearing Techniques for a Sustainable Aquaculture [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/180851 / Compendio

No / Bioactive lipid mediators derived from polyunsaturated fatty acids (PUFA) and exhibit a range of tissue and cell-specific activities in many physiological and pathological processes. Electrospray tandem mass spectrometry coupled to liquid chromatography (LC/ESI-MS/MS) is a sensitive, versatile analytical methodology for the qualitative and quantitative analysis of lipid mediators. Here we present an LC/ESI-MS/MS assay for the simultaneous analysis of twenty mono- and poly-hydroxy fatty acid derivatives of linoleic, arachidonic, eicosapentaenoic and docosahexaenoic acids. The assay was linear over the concentration range 1-100 pg/¿L, whilst the limits of detection and quantitation were 10-20 and 20-50 pg respectively. The recovery of the extraction methodology varied from 76-122% depending on the metabolite. This system is useful for profiling a range of biochemically-related potent mediators including the newly discovered resolvins and protectins, and their precursor hydroxy-eicosapentaenoic and hydroxy-docosahexaenoic acids, and, consequently, advance our understanding of the role of PUFA in health and disease. / Wellcome Trust, British Heart Foundation

Le récepteur LSR est un acteur important du métabolisme hépatique, puisqu'il joue un rôle dans la clairance des lipoprotéines à ApoB/ApoE riches en triglycérides durant la période postprandiale. Dans cette étude, nous avons montré qu'un traitement in vitro par DHA peut augmenter les niveaux de protéine et d'activité LSR dans les cellules d'hépatome de souris Hepa 1-6. En toute cohérence, un régime supplémenté en DHA a conduit à élever les niveaux de protéine LSR hépatique chez la souris. Mais aucune de ces deux études n'a montré de changement au niveau des ARNm. Ceci suggère que l'enrichissement en DHA influe positivement sur le microenvironnement de LSR et son ancrage à la surface de la cellule. Nous avons ensuite étudié le rôle du récepteur PPAR[alpha] dans la régulation du gène lsr. Une analyse in silico nous a permis d'identifier des éléments PPRE dans la région 5' régulatrice du gène humain et de ses homologues de souris et de rat. Des traitements pharmacologiques par des agoniste et antagoniste spécifiques de PPAR[alpha] ont montré que ce récepteur est impliqué dans la régulation transcriptionnelle de l'expression du LSR dans les cellules Hepa 1 6. Enfin, une analyse transcriptomique a révélé une diminution de l'expression de PPAR[alpha] et d'autres gènes impliqués dans le métabolisme lipidique hépatique chez la souris LSR+/- sous régime standard ou riche en graisses. En conclusion, toutes ces études indiquent que l'activité LSR hépatique est sous le contrôle de facteurs nutritionnels capables d'activer divers mécanismes de régulation, faisant du LSR une cible d'intérêt potentiel pour des stratégies nutritionnelles ou thérapeutiques destinées à prévenir ou traiter les dyslipidémies / Lipolysis stimulated lipoprotein receptor (LSR) plays an important role in the clearance of ApoB/ApoE containing triglyceride-rich lipoproteins during postprandial phase. In this study, we demonstrated that in vitro treatment of mouse hepatoma cells, Hepa 1-6, with docosahexaenoic acid (DHA) led to an increase in LSR protein levels as well as its activity. Furthermore, the mice placed on the diet supplemented with DHA showed an increase in hepatic LSR protein. However, the mRNA levels remained unchanged in both in vitro and in vivo studies, suggesting that DHA enrichment may result in changes in LSR microenvironment that could affect its anchorage at the surface of cell membrane. Specific peroxisome proliferator response elements were identified in the upstream region of human, mouse and rat lsr gene by in silico analysis. We therefore sought to determine the role of the transcription factor, peroxisome proliferator-activated receptor (PPAR[alpha]), in LSR regulation. In vitro pharmacological studies using PPAR[alpha]-selective agonist and antagonist agents demonstrated that PPAR[alpha] is indeed involved in the transcriptional regulation of LSR expression. Furthermore, qPCR array analysis revealed the downregulation of PPAR[alpha] and various genes involved in hepatic lipid metabolism in LSR+/- mice on standard and high-fat diets. In conclusion, these studies show that the hepatic LSR activity is controlled by dietary factors that can activate various pathways involved in regulating lipid homeostasis, therefore representing LSR as a potential target for either nutritional or therapeutic strategies towards the prevention or treatment of dyslipidemia

L’optimisation du statut tissulaire en acides gras polyinsaturés à longues chaines n-3 (AGPILC n-3) fait partie des objectifs actuels en nutrition humaine. En effet, de nombreux effets bénéfiques sur la santé humaine ont été démontrés pour ces différents AGPILC n-3. Néanmoins, la bioconversion de ces dérivés à partir de leur précurseur alimentaire, l’acide α-linolénique, est limitée chez l’Homme et ne permet pas de subvenir aux besoins. Parmi cette famille d’acides, l’acide docosahexaénoïque (DHA) et de l’acide eicosapentaénoïque (EPA) ont été largement étudiés à l’inverse de ceux de l’acide docosapentaénoïque n-3 (DPA). Le premier objectif de ces travaux a été d’étudier l’impact d’une supplémentation nutritionnelle en DPA sur le métabolisme des acides gras et le métabolisme lipidique chez le rat sain. Pour cela, une méthode de purification du DPA à haute pureté et en grande quantité par chromatographie liquide a tout d’abord été mise en place. Ce résultat participe à faciliter la recherche future des effets in vivo du DPA dont la disponibilité commerciale est faible. Après sa supplémentation nutritionnelle, le DPA a été incorporé dans de multiples tissus et a impacté les compositions tissulaires en AGPILC n-3 de certains organes spécifiques comme le foie, le cœur, le poumon, la rate et le rein. Ainsi, la supplémentation en DPA pourrait être une source d’EPA, de DPA, et dans une moindre mesure de DHA. Enfin, comparé à une supplémentation en EPA ou en DHA, le DPA a été l’AGPILC n-3 le plus puissant pour améliorer la triglycéridémie et la cholestérolémie des animaux. Ces informations participeront certainement à une prise en compte plus importante du DPA tant dans les études nutritionnelles ainsi que dans l’installation des futures recommandations nutritionnelles. Le deuxième axe de recherche a porté sur la capacité d’une incorporation partielle de lipides laitiers dans le régime à augmenter le statut tissulaire en AGPI-LC n-3. L’incorporation de lipides laitiers a augmenté le statut en DHA cérébral et rétinien et le statut en DPA dans les autres tissus étudiés. De plus, l’induction de la conversion des AGPILC n-3 à partir de leur précurseur par les lipides laitiers a été complémentaire avec la supplémentation en DPA pour augmenter le statut tissulaire en AGPI-LC n-3. Ces résultats s’insèrent dans un contexte de nutrition infantile, période pendant laquelle l’incorporation des AGPI-LC n-3 est maximale dans les tissus nerveux. Ainsi, ces résultats apportent des données intéressantes tant pour les entreprises que pour les professionnels de santé pour recommander à la consommation des formules infantiles contenant des lipides laitiers. / The optimization of tissue status in n-3 long-chain polyunsaturated fatty acids (n-3 LCPUFA) is one of the current goals in human nutrition. Indeed, many beneficial effects on human health have been demonstrated for these different n-3 LCPUFA. However, the bioconversion of these derivatives from their food precursor, α-linolenic acid, is limited in humans and it does not meet the needs of the organism. Among this family of fatty acids, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) have been widely studied in contrast to those of n-3 docosapentaenoic acid (DPA). The first objective of this work was to study the impact of dietary supplementation with DPA on fatty acid metabolism and lipid metabolism in healthy rats. For this, a method for purifying DPA with high purity and in large quantities by liquid chromatography was first established. This result helps future research of the in vivo effects of DPA, poorly commercially available. After its nutritional supplementation, DPA was incorporated into multiple tissues and it has affected the n-3 LCPUFA tissue compositions of specific organs such as the liver, heart, lung, spleen and the kidney. Thus, supplementation with DPA could be a source of EPA, DPA, and to a lesser extent DHA. Finally, compared to supplementation with EPA or DHA, DPA was the most potent n-3 LCPUFA to improve triglyceridemia and cholesterolemia in animals. This information will certainly contribute to a greater consideration of DPA both in nutritional studies and in the direction of future nutritional recommendations. The second area of ​​research focused on the ability of a partial incorporation of dairy lipids in the diet to increase the tissue status in n-3 LCPUFA. Incorporation of dairy lipids in the diet increased DHA status in brain and retina and DPA status in the other studied tissues. In addition, the induction of conversion of n-3 LCPUFA from their precursor by dairy lipid was complementary with the DPA supplementation to increase n-3 LCPUFA status in tissues. These results fit into a context of infant nutrition, during which the incorporation of n-3 LCPUFA is maximal in nervous tissues. Thus, these results provide interesting data for both companies and health professionals to recommend infant formulas containing dairy lipids for consumption.

Human skin has distinct lipid metabolism and production of bioactive lipid mediators that can be modulated by nutritional supplementation with omega-3 polyunsaturated fatty acids (n-3 PUFA), of which eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids exert anti-inflammatory effects. The aims of this project were to gain better understanding of their individual mechanisms in human epidermis and dermis. HaCaT keratinocytes, 46BR.1N fibroblasts, primary human epidermal keratinocytes and dermal fibroblasts were treated with EPA or DHA for 72h and then sham-irradiated or exposed to 15 mJ/cm2 ultraviolet radiation (UVR). Viability was measured by the MTT assay. The expression of cyclooxygenase-2 (COX-2), microsomal prostaglandin synthase-1 (mPGES-1) and 15-hydroxyprostaglandin dehydrogenase (15-PGDH) proteins was explored by western blotting. Human skin explants (n=4 donors) were cultured for 3 or 6 days and supplemented with EPA, DHA or vehicle. Culture media were collected to evaluate tissue damage and PUFA cytotoxicity (lactate dehydrogenase assay). Epidermal and dermal lipid profiles were assessed by gas chromatography and liquid chromatography coupled to tandem mass spectrometry. Primary keratinocytes were treated with fatty acids and various lipid mediators for 48h. Their effect was determined by the scratch assay and transepithelial electrical resistance. UVR upregulated COX-2 in HaCaT and primary epidermal keratinocytes, but did not affect mPGES-1 and 15-PGDH protein expression. UVR upregulated COX-2 and mPGES-1 in 46BR.1N fibroblasts but had no effect on 15-PGDH expression. The same UVR dose did not alter the expression of COX-2, mPGES-1 and 15-PGDH in primary dermal fibroblasts. Only EPA attenuated COX-2 expression in HaCaT and primary keratinocytes and either EPA or DHA had any effect in 46BR.1N and primary fibroblasts. Skin explants showed initial post-biopsy tissue damage. EPA and DHA supplementation augmented cellular levels of the corresponding fatty acids in both epidermis and dermis to a different extent. Increased uptake of DHA in the dermis was accompanied by reduced arachidonic acid levels. EPA treatment stimulated the production of PGE3 and various HEPE in epidermis, while DHA treatment caused high levels of HDHA species in dermis. N-3 PUFA and their derivatives delayed wound healing, cell migration and epidermal barrier permeability, while n-6 PUFA lipids showed the opposite effect. Overall, these findings suggest that EPA and DHA differently affect skin cells and skin, with EPA preference in epidermis and DHA in the dermis. These results highlight the importance of differential skin responses that could be important in skin health and disease.

Un des phénomènes sociétaux marquants de ces dernières années est le vieillissement de la population et en conséquence, une hausse considérable du nombre de personnes âgées. Dans ce contexte, la recrudescence des pathologies chroniques liées au vieillissement, dont la maladie d’Alzheimer, est devenue un enjeu majeur de santé publique. L’impact de nombreux facteurs environnementaux modulables, l’aspect chronique et évolutif des mécanismes pathogènes mis en jeu, doivent inciter à développer des interventions préventives permettant de minimiser les risques de développer ces maladies liées au vieillissement. Ce travail nous a permis de mettre en évidence l’importance d’utiliser des modèles d’étude et des modes d’expérimentation adaptés au vieillissement pour espérer en ralentir ou retarder les processus délétères. Nos résultats ont aussi permis d’identifier les membranes comme des éléments essentiels au bon fonctionnement cérébral. L’altération de la composition et de l’architecture des membranes neuronales chez la souris âgée perturbe leurs fonctionnalités et diminue les capacités de réponse neuroprotectrices recherchées notamment lors des thérapies anti-Alzheimer. Nous avons aussi observé des modifications membranaires comparables chez les souris rendues dyslipidémiques par un régime alimentaire excessif en lipides saturés auquel nous avons pu clairement attribuer un rôle pro-vieillissement. Nous avons finalement démontré le potentiel préventif d’une supplémentation alimentaire en acide docosahexaénoïque, l’acide gras polyinsaturé à longue chaîne majoritaire dans le cerveau, et pu conclure en sa capacité de restaurer une réponse neuroprotectrice altérée chez la souris âgée / One of the marked societal phenomena in recent decades is the aging of populations due to continually increasing lifespans and as a result, a considerable surge in the number and proportion of elderly, particularly in Western countries. In this demographic context, the rise of chronic diseases related to aging, including Alzheimer’s disease and other types of dementia, has become a major public health issue. The impact of modifiable environmental factors, evolution of the pathogenic mechanisms involved, and the lack of curative treatments illustrates the need for the development of interventions to prevent or delay the onset of these aging-related diseases. The present work demonstrates the importance of using age-adapted study models and experimental methods with the goal towards slowing or delaying age-related deleterious processes. Secondly, our results have identified membranes as an essential part for normal brain function. The composition and architectural changes in the neuronal membranes of elderly mice disrupt their functionality and reduce neuroprotective responsiveness such as those sought by anti-Alzheimer’s therapies. We also observed similar pro-aging-type changes in brain membranes of dyslipidemic mice fed a high-fat diet. Thus, disturbances of lipid homeostasis are correlated with an increased risk of developing aging-related cardiovascular and metabolic as well as neurodegenerative diseases. We finally demonstrated the preventive potential of dietary supplementation with docosahexaenoic acid, the most abundant long-chain polyunsaturated fatty acid in the brain, and observed its ability to restore a neuroprotective response that was impaired in older mice

Indiana University-Purdue University Indianapolis (IUPUI) / Cellular membranes contain a staggering diversity of lipids. The lipids are heterogeneously distr ibuted to create regions, or domains, whose physical properties differ from the bulk membrane and play an essential role in modulating the function of resident proteins. Many basic questions pertaining to the formation of these lateral assemblies remain. T his research employs model membranes of well - defined composition to focus on the potential role of polyunsaturated fatty acids (PUFAs) and their interaction with cholesterol (chol) in restructuring the membrane environment. Omega - 3 (n - 3) PUFAs are the main bioactive components of fish oil, whose consumption alleviates a variety of health problems by a molecular mechanism that is unclear. We hypothesize that the incorporation of PUFAs into membrane lipids and the effect they have on molecular organization may be, in part, responsible. Chol is a major constituent in the plasma membrane of mammals. It determines the arrangement and collective properties of neighboring lipids, driving the formation of domains via differential affinity for different lipids . T he m olecular organization of 1 -[ 2 H 31 ]palmitoyl -2- eicosapentaenoylphosphatidylcholine (PEPC - d 31 ) and 1 -[ 2 H 31 ]palmitoyl -2- docosahexaenoylphosphatidylcholine (PDPC -d 31 ) in membran es with sphingomyelin (SM) and chol (1:1:1 mol) was compared by solid - state 2 H NMR spectroscopy. Eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids are the two major n - 3 PUFAs found in fish oil, while PEPC -d 31 and PDPC -d 31 are phospholipids containing the respective PUFAs at the sn - 2 position and a perdeuterated palmitic acid a t the sn - 1 position . Analysis of s pectra recorded as a function of temperature indicate s that in both cases, formation of PUFA - rich (less ordered) and SM - rich (more ordered) domains occurred. A surprisingly substantial proportion of PUFA was found to infil trate the more ordered domain. There was almost twice as much DHA (65%) as EPA (30%) . The implication is that n - 3 PUFA s can incorporate into lipid rafts, which are domains enriched in SM and chol in the plasma membrane, and potentially disrupt the activity of signaling proteins that reside therein. DHA, furthermore, may be the more potent component of fish oil. PUFA - chol interactions were also examined through affinity measurements. A novel method utilizing electron paramagnetic resonance (EPR) was develope d, to monitor the partitioning of a spin - labeled analog of chol , 3β - doxyl - 5α - cholestane (chlstn), between large unilamellar vesicles (LUVs) and met hyl - β - cyclodextrin (mβCD). The EPR spectra for chlstn in the two environments are distinguishable due to the substantial differences in tumbling rates , allowing the population distribution ratio to be determined by spectral simulation. Advantages of this approach include speed of implementation and a vo idance of potential artifact s associated with physical separation of LUV and mβCD . Additionally, in a check of the method, t he relative partition coefficients between lipids measured for the spin label analog agree with values obtained for chol by isothermal titration calorimetry (ITC). Results from LUV with different composition confirmed a hierarchy of decreased sterol affinity for phospholipids with increasing acyl chain unsaturation , PDPC possessing half the affinity of the corresponding monounsaturated phospholipid. Taken together, the results of these studies on model membranes demonstrate the potential for PUFA - driven alteration of the architecture of biomembranes, a mechanism through which human health may be impacted.

No description available.