Vortex Tube (VT) is a simple device having no moving mechanical parts, in which compressed gas at high pressure is injected through one or more tangential nozzles into a vortex chamber resulting in the separation of the inlet flow into two low pressure streams. One of the streams is the peripheral flow that is warmer than the inlet stream while the other is the central (core) flow that is colder than the inlet stream. This separation of the inlet flow into high and low temperature streams is known as temperature or energy separation. It is suggested by many investigators that compressed air of few atmospheres pressure and at room temperature can produce temperatures as high as +200ºC at the hot end (peripheral flow exit) and as low as -50ºC at the cold end (core flow exit) of the VT. Though VTs have large potential for simple heating and cooling applications, the mechanism of energy separation is not clear so far. Based on their studies, many investigators have suggested various theories, different from each other, but having specific lacunas and is an unresolved issue. Also, till date, experimental and industrial designs of the VTs are based purely on empirical correlations. Apart from heating and cooling applications, VTs can also be used for separation of binary gas mixtures and separation of oxygen from two-phase precooled air stream. The conceptual futuristic cryogenic launch vehicle designs are being attempted with in-flight liquid oxygen (LOX) collection system that significantly improves the pay load fraction. Vortex tube technology is one of the few promising technologies for futuristic in-flight LOX separation based launch vehicles. This technology has significant advantages over its counterparts as it is a simple, compact and light weight, and most importantly have no moving parts and unaffected by gravity and orientation. In order that VTs become an acceptable technology for in-flight LOX separation system, it is necessary to achieve minimum oxygen purity of 90% with more than 60% yield (separation efficiency) for the oxygen enriched stream in the VT. A survey of the available open literature has shown very little reported details, in particular, on achieving the required specifications for in-flight LOX separation systems. Till date, the highest LOX purity of 60% with 40% separation efficiency has been reported with VT technology. In view of the above mentioned facts, the work carried out has been focused on to: • Optimize the critical parameters of the VT to achieve maximum energy separation by CFD and experimental studies. • Understand the flow behaviour in the VT by estimating the velocity, temperature and pressure profiles at various locations in the VT and validation of secondary circulation flow and its effect on the performance of energy separation in VT. • Estimation of the energy transfer between the core and the peripheral layers of fluid flow in VT by analytical and CFD methods to propose the most appropriate mechanism of energy separation in VT. • Design and development of a dedicated experimental setup for both energy separation and LOX separation studies in VTs. • Design and fabrication of straight and conical VTs and experimental programme on energy separation and LOX separation. • Development of the VT air separation technology to achieve the required specifications of in-flight LOX separation system for futuristic launch vehicles. With these specific objectives and motivations, the total work was carried out with the following planned and sequential steps: • The first step was the CFD modeling of the VT with the available CFD software (Star-CD) and obtain the energy separation phenomena for a 12mm diameter VT. After gaining sufficient confidence level, optimization of the critical parameters like the air injection nozzle profile, number of nozzles, cold end orifice diameter dc, length to diameter (L/D) ratio, hot gas fraction etc of the VT was carried out through CFD and experimental studies. • The studies show that 6 convergent nozzles perform better in comparison to other configurations like circular helical, rectangular helical, 2 convergent and 6 straight nozzles. The studies also show that cold end orifice diameter (dc) plays an important role on energy separation and bring out the existence of secondary circulation flow with improper design of cold end orifice diameter. Through our studies, the effect of cold end diameter on the secondary circulation flow has been evaluated for the first time. Also, the mechanism of energy transfer in VT based on heat pump mechanism enabled by secondary circulation flow as suggested by some investigators has been evaluated in our studies. The studies show that cold end orifice diameter dc = 7mm is optimum for 12mm diameter VT, which matches fairly with the correlations given by other investigators. The studies confirms that CFD modeling carried out in this work is capable of selecting the correct dc value for a VT, without resorting to the empirical correlations as a design guide or a laborious experimental programme. • Through the CFD and experimental studies on different length to diameter (L/D) ratios and hot gas fractions, maximum hot gas temperature of 391K was obtained for L/D = 30 with hot gas fraction of 12-15 % and minimum cold gas temperature of 267K for L/D = 35 was obtained for cold gas fraction ≈ 60% (lowest cold gas fraction possible with the present experimental system). • CFD analysis has been carried out to investigate the variation of static and total temperatures, static and total pressures as well as the velocity components of the particles as it progresses in the flow field, starting from the entry through the nozzles to the exit of the VT by tracking the particles to understand the flow phenomenon and energy transfer mechanism inside the VT. The studies indicate that the mechanism of energy transfer from the core flow to the peripheral flow in VT is predominantly occurs by the tangential shear work. Thus the investigations reported in the thesis have given a clear understanding of the contributing mechanism for energy separation in VT, which has been an unresolved issue for long time. The net energy transfer between the core and the peripheral fluid has been calculated analytically and compared with the values obtained by CFD model for VTs of L/D ratios equal to 10 and 30. The net energy transfer by analytical and CFD model for VT with L/D = 10 is 159.87W and 154.2W respectively whereas the net energy transfer by analytical and CFD model for VT with L/D = 30 is 199.87W and 192.3W respectively. The results show that CFD results are in very good agreement with the analytical results and CFD can be used as a tool for optimization of the critical parameters and to analyze the flow parameters and heat transfer analysis for VTs. Also, the net energy transfer between the core and peripheral fluids calculated analytically matches very well with that of the net energy transfer by CFD analysis, without considering the effect of acoustic streaming. Thus acoustic streaming may not be the mechanism of energy separation in VT as suggested by some investigators. • By optimizing the critical parameters of the 12mm diameter straight VT through CFD and experimental studies, LOX separation studies have been carried out using both straight and conical VTs of dc = 7mm and of different L/D ratios for high LOX purity and separation efficiency. It is observed that conical (3º divergence) VTs perform better as compared to straight VTs for LOX separation whereas straight VTs perform better for energy separation. The better performance of conical VT as compared to straight VTs can be attributed to its increased surface area for condensation-evaporation phenomenon of oxygen and nitrogen molecules. Experimental studies have been conducted to evaluate the influence of the inlet pressure and the inlet temperature (liquid fraction) on LOX purity. Studies indicate that for achieving high LOX purity for the studied experimental system, the inlet pressure is to be in the range of 6-6.5bar and there exists a very narrow band of inlet temperature zone in which high LOX purity can be achieved. Experimental studies on VTs show that VT can be optimized suitably either for high LOX purity with low separation efficiency or low LOX purity with high separation efficiency by adjusting the hot end mass fraction accordingly. It is also observed that it is not possible to obtain both high purity and high separation efficiency simultaneously with the single VT. Staging approach has to be adapted to achieve higher LOX purity with higher separation efficiency. By staging the VTs, the enriched air stream (hot end outlet flow) from the first stage of VTs is introduced to the inlet of the second stage of VTs. Experimental studies have been conducted to evaluate the design parameters on staging of VTs. LOX purity of 48% with 89% separation efficiency has been achieved for conical first stage VT of L/D = 25. LOX purity of about 94% with separation efficiency of 84% has been achieved for 50% oxygen content at the inlet of the second stage VT. Similarly, LOX purity of 96% with separation efficiency of 73.5% has been achieved for 60% oxygen content at the inlet of the VT. This is the highest LOX purity and separation efficiency reported so far indicating that, conical VT of optimized diameter, L/D ratio and orifice diameter can yield the hot end flow very close to the target value of futuristic in-flight LOX separation based launch vehicles. The present investigation has focused the optimization of the critical parameters of VTs through CFD and experimental studies. It has also given an insight to the mechanism of energy transfer between the core and peripheral flow in VT by evaluating two of the existing theories on mechanism of energy transfer in VT. The studies also highlighted the fact that custom designed and precision fabricated VTs can be very useful for obtaining maximum / minimum temperatures of fluid flow as well as LOX separation with high purity and high separation efficiency needed for futuristic in-flight LOX separation based space launch vehicles.

Atrial natriuretic factor (ANF) is a polypeptide hormone produced in heart atria, stored in atrial secretory granules and released into the circulation in response to various stimuli. Proper sorting of ANF at the level of the trans-Golgi network (TGN) is required for the storage of ANF in these specific granules, and this sorting of hormones has been found to be associated with G-proteins. Specifically, the Go protein alpha-subunit (Gαo) was established to participate in the stretch-secretion coupling of ANF, but may also be involved in the transporting of ANF from the TGN into atrial granules for storage and maturation. Based on knowledge of Gαo involvement in hormone production in other endocrine tissues, protein-protein interactions of Gαo and proANF and their immunochemical co-localization in granules, the direct involvement of these two proteins in atrial granule biogenesis is probable. In this study, mice were created using the Cre/lox recombination system with a conditional Gαo knockout in cardiocytes to study and characterize ANF production, secretion and granule formation. Deletion of this gene was successful following standard breeding protocols. Characterization and validation of cellular and molecular content of the knockout mice through mRNA levels, protein expression, peptide content, electron microscopy, and electrocardiography determined that a significant phenotypic difference was observed in the abundance of atrial granules. However, Gαo knockout mice did not significantly alter the production and secretion of ANF and only partially prevented granule biogenesis, likely due to incomplete Gαo knockout. These studies demonstrate an involvement of Gαo in specific atrial granule formation.

This study is focused on study of changes of enzyme and low-molecular weight antioxidants in different fruits during long-term storage. In theoretical part individual low-molecular weight antioxidants and enzymes are described. The main causes of fruit decay and some possibilities of fruit preservation and storage are summarized. As biological material some common fruits were chosen - green and red apples, peaches, plums and white grapes. The fruits were stored in laboratory, cellar, in refrigerator and in freezer. In freezing experiments some ways of fruit preparation and processing were tested and their influence on fruit antioxidant status was compared. Shortened storage experiment was applied on blueberries, cranberries, raspberries and strawberries too. In fruits some group parameters – total antioxidant status, dry mass content, ascorbate level, total flavonoids and total phenolics were analyzed spectrophotometrically. Individual flavonoids and phenolics were determined by RP-HPLC/UV-VIS and on-line LC/PDA/ESI-MS. Antioxidant enzyme activities (superoxide dismutase SOD, catalase CAT, polyphenol oxidase PPO and lipoxygenase LOX) were measured by spectrophotometry. The surface microscopy and cultivation of moulds from fruit surface were performed too. Influence of storage conditions on biological activities is dependent on fruit sort. Freezing is the most suitable procedure for long-term storage without significant changes of active substance content. Long-term storage in controlled temperature conditions and/or atmosphere is usable for fruits with longer storage period. In these fruits stabile levels of antioxidant enzymes are stored for relatively long time. Some of enzymes act synergistically. Enzyme activities differed according to storage phase; at the beginning mainly high SOD and LOX activities were observed. CAT and PPO are probably activated as defence systems in rippened and/or damaged fruits. Levels of total as well as individual low molecular weight antioxidants varied during storage in all sorts, generally, increased course with longer storage period can be observed.

Les lymphocytes T CD8+ jouent un rôle prépondérant dans le contrôle de l’infection par le VIH suggérant ainsi qu’un vaccin efficace doit induire une réponse cellulaire T CD8+ robuste et durable.En fournissant une source antigénique importante qui fait défaut chez les patients répondeurs aux poly-chimiothérapies antirétrovirales, la vaccination thérapeutique pourrait favoriser ce type de réponse cellulaire, non restaurée par les seuls traitements antirétroviraux. Les cellules dendritiques (DC) autologues, qu’elles soient chargées en virus inactivé, en peptides viraux ou en ARN messagers (ARNm) sont capables d’induire des réponses robustes des cellules T CD8+ contre les antigènes du VIH, faisant de bons candidats vaccins thérapeutiques.Dans ce projet, nous avons étudié l’efficacité d’une vaccination thérapeutique basée sur l’injection de DC autologues transfectées ex vivo avec des ARNm codant les protéines du VIH (projet initié en collaboration avec Guido Vanham, Institute of Tropical Medicine, Anvers). La composante antigénique du vaccin est fournie par les séquences virales endogènes du patient; ainsi nous espérons adapter la spécificité de la réponse immunitaire aux virus autologues. Nous travaillons avec le Macaque cynomolgus comme modèle animal d’infection par les virus d’immunodéficience (SIV), modèle pertinent dans l’approfondissement des connaissances en pathogénèse et en développement vaccinal, notamment pour tester l’innocuité et l’efficacité des vaccins basés sur l’utilisation des DC. La mise au point un système de culture de DC basé sur la prolifération puis la différenciation des progéniteurs hématopoïétiques CD34+, qui nous a permis de générer in vitro des DC matures en nombre suffisant pour la vaccination (au moins 10.106).Nous avons caractérisé ces DC phénotypiquement, avec une attention particulière vis-à-vis de l’expression de lectines de type C et d’autres récepteurs DC-spécifiques. Nous avons mis en évidence que l’état d’activation des DC définit le niveau d’expression de ces molécules. Ces récepteurs membranaires sont impliqués dans la capture et la présentation des antigènes aux effecteurs, ainsi que dans l’orientation de la réponse immune. Le phénotype des DC générées in vitro a été comparé au phénotype de plusieurs population de cellules présentatrices d’antigène présentes in vivo chez le macaque.La transfection avec des ARNm par électroporation est un moyen sûr et efficace de charger les DC en antigènes. Nous avons étudié dans un premier temps l’immunogénicité du vaccin chez des macaques sains. Les résultats montrent qu’une réponse cellulaire de type Th1 (IFN-gamma et interleukine-2) spécifique à Gag est induite dès la première injection et peut être renforcée après les injections suivantes. Nous avons également mis en évidence l’induction de lymphocytes T CD8+ capables de sécréter en réponse à Gag plusieurs de ces cytokines, IFNg, IL2, TNFa, la chimiokine MIP1b, ou encore le marqueur de dégranulation cytotoxique CD107a. En revanche, aucune sécrétion de cytokine de type 2 ni de réponse anticorps spécifique de Gag n'a pu être mise en évidence.La seconde partie de notre projet a consisté en l’étude d’une stratégie de vaccination par ciblage des DC in vivo avec des protéines de fusion DC-spécifiques associées à des antigènes. Ce travail a été mené en collaboration avec le Baylor Insitute for Immunology Research (BIIR) à Dallas. Nous avons tout d’abord sélectionné les meilleurs clones d’Ac spécifiques de molécules humaines pour leur réactivité croisée avec les molécules du macaque cynomolgus. L’immunogénicité de protéines de fusion (dirigées contre LOX-1 et DC-ASGPR couplées à Gag du VIH ou HA1 de Influenza), a ensuite été évaluée in vivo chez le macaque. Ces travaux ont confirmé in vivo que l’induction de différents profils de réponses immunitaires Ag-spécifiques (Th1 et IL10) est possible par ciblage in vivo des DC avec des protéines de fusion dirigées contre différentes molécules (LOX-1 et DC-ASGPR). / Current treatments against HIV infection would benefit from the development of complementary immunotherapeutic strategies. Dendritic cells (DC) play a major role in the induction of immune responses. In this thesis, we examine the use of the DC in therapeutic vaccination. Ex vivo loading of DC with messenger RNA encoding viral proteins and in vivo loading of DC by targeting antigen to DC-specific endocytic receptors are two strategies evaluated in healthy macaque.

The present study aims at the isolation and identification of diverse phenolic polyketides from Aloe vera (L.) Burm.f. and Aloe plicatilis (L.) Miller and includes their 5-LOX/COX-1 inhibiting potency. After initial Sephadex-LH20 gel filtration and combined silica gel 60- and RP18-CC, three dihydroisocoumarins (nonaketides), four 5-methyl-8-C-glucosylchromones (heptaketides) from A. vera, and two hexaketide-naphthalenes from A. plicatilis have been isolated by means of HSCCC. The structures of all polyketides were elucidated by ESI-MS and 2D 1H/13C-NMR (HMQC, HMBC) techniques. The analytical/preparative separation of 3R-feralolide, 3′-O-β-d-glucopyranosyl- and the new 6-O-β-d-glucopyranosyl-3R-feralolide into their respective positional isomers are described here for the first time, including the assignment of the 3R-configuration in all feralolides by comparative CD spectroscopy. The chromones 7-O-methyl-aloesin and 7-O-methyl-aloeresin A were isolated for the first time from A. vera, together with the previously described aloesin (syn. aloeresin B) and aloeresin D. Furthermore, the new 5,6,7,8-tetrahydro-1-O-β-d-glucopyranosyl- 3,6R-dihydroxy-8R-methylnaphtalene was isolated from A. plicatilis, together with the known plicataloside. Subsequently, biological-pharmacological screening was performed to identify Aloe polyketides with anti-inflammatory potential in vitro. In addition to the above constituents, the anthranoids (octaketides) aloe emodin, aloin, 6′-(E)-p-coumaroyl-aloin A and B, and 6′-(E)-p-coumaroyl-7-hydroxy-8-O-methyl-aloin A and B were tested. In the COX-1 examination, only feralolide (10 µM) inhibited the formation of MDA by 24%, whereas the other polyketides did not display any inhibition at all. In the 5-LOX-test, all aloin-type anthranoids (10 µM) inhibited the formation of LTB4 by about 25–41%. Aloesin also displayed 10% inhibition at 10 µM in this in vitro setup, while the other chromones and naphthalenes did not display any activity. The present study, therefore, demonstrates the importance of low molecular phenolic polyketides for the known overall anti-inflammatory activity of Aloe vera preparations.

This thesis contains the results of optical flame emission measurements of the Vulcain 2.1engine and the plasma emission spectroscopy of the Lumen Project engine. The plume spectroscopyis analyzed, ordered and studied in detail to offer the best possible molecular composition.The main focus relied on the hydroxide radical, blue radiation and other moleculesanalysis of the intensities encountered during the tests. The plasma emission spectroscopy isfocused on the determination of the plasma temperature value in LIBS measurements. Thehydrogen plasma temperature determination of the local thermodynamic equilibrium, followedby the carbon and sequentially oxygen plasma is obtained. The quality of the LTE isto be determined to judge the truthworthness of the determined temperatures. Both the testsare analyzed thanks to the use of spectrographs, cameras and dedicated software for opticalapplications. The results related to the Vulcain 2.1 LOX/LH2 engine showed the evolutionof the plume in different ROF or pressure variations. Furthermore, the results of the LumenProject LOX/methane engine led to the determination of the plasma temperatures and a firstestimation of the LTE quality. / Die vorliegende Arbeit präsentiert die Ergebnisse der Abgasstrahlspektroskopie des H2/LOXVulcain 2.1 Triebwerks und der Zündplasma Spektroskopie des CH4/LOX Triebwerks desLUMEN Projektes. Die Abgasstrahlspektroskopie wurde analysiert und im Detail untersuchtum die am besten passende molekulare Zusammensetzung herauszuarbeiten. DasHauptaugenmerk liegt dabei auf dem Hydroxyl- Radikal, der Blauen Strahlung und molekularerIntensitätsanalyse. Bei der Zündplasmaanalyse liegt der Fokus auf der Bestimmungdes LTE Zustands (Lokales thermodynamisches Gleichgewicht) in LIBS. Die Temperaturdes Wasserstoff-, Kohlenstoff und Sauerstoffplasmas wird herangezogen, um die Qualitätdes LTE Zustands zu beurteilen. Für die Testdurchführung wurden Spektrographen, Kamerasund bestimmte Auswertungstools für optische Anwendungen benutzt. Das Verhaltendes Vulcain 2.1 Abgasstrahls abhängig von verschiedenen ROF und Druckstufen ist in denErgebnissen beschrieben. Für das LUMEN Triebwerk konnten erste Zündplasmatemperaturenbestimmt werden und geben einen Rückschluss auf die Qualität des LTE.

Les maladies cardiovasculaires (MCV) sont la principale cause de décès dans les pays occidentaux et constituent la principale complication associée au diabète. La lipoprotéine lipase (LPL) est une enzyme clé du métabolisme des lipides et est responsable de l'hydrolyse des lipoprotéines riches en triglycérides (TG). Plusieurs études ont démontré que la LPL sécrétée par les macrophages dans la paroi artérielle est pro-athérogénique. La dysfonction endothéliale caractérise les stades précoces du processus athérosclérotique. Il a été observé qu’un récepteur nouvellement identifié des lipoprotéines de basse densité oxydées (LDLox), le récepteur de type lectine des LDLox (LOX-1), est fortement exprimé dans les lésions athérosclérotiques humaines et dans l’aorte de rats diabétiques, suggérant un rôle clé de LOX-1 dans la pathogénèse de l’athérosclérose diabétique. Au vu du rôle potentiel de la LPL macrophagique et du LOX-1 dans l’athérosclérose associée au diabète de type 2, nous avons évalué la régulation de ces deux molécules pro-athérogéniques par des facteurs métaboliques et inflammatoires augmentés dans le diabète, soit la leptine, l’acide linoléique (LA) et la protéine C-réactive (CRP). Nos résultats démontrent que : 1) Dans les cellules endothéliales aortiques humaines (HAECs), LA augmente l’expression protéique de LOX-1 de façon temps- et dose-dépendante; 2) La pré-incubation de HAECs avec des antioxydants et des inhibiteurs de la NADPH oxydase, de la protéine kinase C (PKC) et du facteur nucléaire-kappa B (NF-kB), inhibe l’effet stimulant de LA sur l’expression protéique de LOX-1; 3) Dans les HAECs traitées avec LA, on observe une augmentation d’expression des isoformes classiques de la PKC; 4) LA augmente de manière significative l’expression génique de LOX-1 ainsi que la liaison des protéines nucléaires extraites des HAECs à la séquence régulatrice NF-kB présente dans le promoteur du gène de LOX-1; 5) LA augmente, via LOX-1, la captation des LDLox par les cellules endothéliales. Pris dans leur ensemble, ces résultats démontrent que LA augmente l’expression endothéliale de LOX-1 in vitro et appuient le rôle clé de LA dans la dysfonction endothéliale associée au diabète. Au vu de nos études antérieures démontrant qu’une expression accrue de LPL macrophagique chez les patients diabétiques de type 2 et que l’augmentation de facteurs métaboliques dans cette maladie, soit l’homocystéine (Hcys), les acides gras et les produits terminaux de glycation (AGE), accroissent l’expression de la LPL macrophagique, nous avons par la suite déterminé l’effet, in vitro, de deux autres facteurs métaboliques et inflammatoires surexprimés dans le diabète, soit la leptine et la CRP, sur l’expression de la LPL macrophagique. Les concentrations plasmatiques de leptine sont élevées chez les patients diabétiques et sont associées à un accroissement des risques cardiovasculaires. Nous avons démontré que : 1) Dans les macrophages humains, la leptine augmente l’expression de la LPL, tant au niveau génique que protéique; 2) L’effet stimulant de la leptine sur la LPL est aboli par la pré-incubation avec un anticorps dirigé contre les récepteurs à la leptine (Ob-R), des inhibiteurs de la PKC et des antioxydants; 3) La leptine augmente l’expression membranaire des isoformes classiques de la PKC et la diminution de l’expression endogène de la PKC, abolit l’effet de la leptine sur l’expression de la LPL macrophagique; 4) Dans les macrophages murins, la leptine augmente le taux de synthèse de la LPL et augmente la liaison de protéines nucléaires à la séquence protéine activée-1 (AP-1) du promoteur du gène de la LPL. Ces observations supportent la possibilité que la leptine puisse représenter un facteur stimulant de la LPL macrophagique dans le diabète. Finalement, nous avons déterminé, in vitro, l’effet de la CRP sur l’expression de la LPL macrophagique. La CRP est une molécule inflammatoire et un puissant prédicteur d’événements cardiovasculaires. Des concentrations élevées de CRP sérique sont documentées chez les patients diabétiques de type 2. Nous avons démontré que : 1) Dans les macrophages humains, la CRP augmente l’expression de la LPL au niveau génique et protéique et la liaison de la CRP aux récepteurs CD32 est nécessaire pour médier ses effets; 2) La pré-incubation de macrophages humains avec des antioxydants, des inhibiteurs de la PKC et de la protéine kinase mitogénique activée (MAPK), prévient l’induction de la LPL par la CRP; 3) La CRP augmente l’activité de la LPL, la génération intracellulaire d’espèces radicalaires oxygénées (ROS), l’expression d’isoformes classiques de la PKC et la phosphorylation des kinases extracellulaires régulées 1/2 (ERK 1/2); 4) Les macrophages murins traités avec la CRP démontrent une augmentation de la liaison des protéines nucléaires à la séquence AP-1 du promoteur du gène de la LPL. Ces données suggèrent que la LPL puisse représenter un nouveau facteur médiant les effets délétères de la CRP dans la vasculopathie diabétique. Dans l’ensemble nos études démontrent le rôle clé de facteurs métaboliques et inflammatoires dans la régulation vasculaire de la LPL et du LOX-1 dans le diabète. Nos données suggèrent que la LPL et le LOX-1 puissent représenter des contributeurs clé de l’athérogénèse accélérée associée au diabète chez l’humain. Mots-clés : athérosclérose, maladies cardiovasculaires, diabète de type 2, macrophage, LPL, cellules endothéliales, LOX-1, stress oxydatif, leptine, LA, CRP. / Atherosclerotic cardiovascular disease is the leading cause of death in western countries and is the major complication of diabetes. Lipoprotein lipase (LPL) is a key enzyme in lipid metabolism, responsible for the hydrolysis of triglyceride (TG) rich lipoproteins. Many studies have shown that LPL secreted by macrophages in the arterial wall is proatherogenic. Endothelial dysfunction is a characteristic feature of early-stage atherosclerosis. The observation that lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), a newly identified receptor for oxidized LDL (oxLDL), is highly expressed in human atherosclerotic lesions and upregulated in the aorta of diabetic rats, suggests a key role for LOX-1 in the pathogenesis of diabetic atherosclerosis. Based on the role of macrophage LPL and LOX-1 in atherosclerosis, we sought to investigate the regulation of these two proatherogenic molecules by metabolic and inflammatory factors dysregulated in diabetes namely leptin, linoleic acid (LA) and C-reactive protein (CRP). Our results demonstrated that: 1) In human aortic endothelial cells (HAECs), LA increased LOX-1 protein expression in a time- and dose-dependent manner; 2) Pretreatment of HAECs with antioxidants and inhibitors of NADPH oxidase, protein kinase C (PKC), and nuclear factor-kB (NF-kB) inhibited the stimulatory effect of LA on LOX-1 protein expression; 3) Increased expression of classic PKC isoforms was observed in LA-treated HAECs; 4) LA led to a significant increase in LOX-1 gene expression and enhanced the binding of nuclear proteins extracted from HAECs to the NF-kB regulatory element of the LOX-1 gene promoter; 5) LA enhanced, through LOX-1, oxLDL uptake by endothelial cells. Overall, these results demonstrated that LA enhances endothelial LOX-1 expression in vitro and support a key role for LA in endothelial dysfunction associated with diabetes. Based on our previous studies showing that macrophage LPL expression is increased in patients with type 2 diabetes and that metabolic factors dysregulated in this disease such as glucose, homocysteine (Hcys), fatty acids and advanced glycation end products (AGE), increased macrophage LPL expression, we next determined the effect of two other metabolic and inflammatory factors dysregulated in diabetes, namely leptin and CRP, on macrophage LPL expression in vitro. Leptin levels are elevated in diabetic patients and are associated with greater cardiovascular risks. We found that: 1) In human macrophages, leptin increased LPL expression, at both the gene and protein levels; 2) The stimulatory effect of leptin on LPL was abolished by pre-treatment with anti-leptin receptor (Ob-R) antibody, PKC inhibitors and antioxidants; 3) Leptin increased the membrane expression of conventional PKC isoforms and downregulation of endogenous PKC expression abolished the effects of leptin on macrophage LPL expression; 4) In murine macrophages, leptin raised LPL synthetic rate and enhanced the binding of nuclear proteins to the activated protein-1 (AP-1) sequence of the LPL gene promoter. These observations support the possibility that leptin may represent a macrophage LPL stimulatory factor in diabetes. Finally, we sought to determine the effect of CRP on macrophage LPL expression in vitro. CRP is an inflammatory molecule and a strong predictor of cardiovascular events and high serum levels of CRP are observed in type 2 diabetic patients. We found that: 1) In human macrophages, CRP increased LPL expression at the gene and protein levels and CRP binding to CD32 receptors is required for these effects; 2) Preincubation of human macrophages with antioxidants, PKC and mitogen-activated protein kinase (MAPK) inhibitors, prevented the CRP-induced LPL expression; 3) CRP increased LPL activity, intracellular reactive oxygen species (ROS) generation, classic PKC isozymes expression and extracellular signal-regulated protein kinase (ERK) 1/2 phosphorylation; 4) CRP-treated murine macrophages demonstrated increased binding of nuclear proteins to the AP-1 sequence of the LPL gene promoter. These data suggest that LPL might represent a novel factor underlying the adverse effect of CRP on the diabetic vasculature. Overall, our studies indicate a key role of metabolic and inflammatory factors in the regulation of vascular LPL and LOX-1 in diabetes. Our data suggest that LPL and LOX-1 are key contributors to the accelerated atherogenesis associated with human diabetes. Keywords : atherosclerosis, cardiovascular diseases, type 2 diabetes, macrophage, LPL, endothelial cells, LOX-1, oxidative stress, leptin, LA, CRP.

No description available.

No description available.

For performance optimisation of modern liquid cryogenic bipropellant rocket combustion chambers, one component which plays an important role in reducing the wall side heat flux, is the behaviour of the cooling film. At the Institute of Space Propulsion of the German Aerospace Center (DLR) in Lampoldshausen, hot test runs have been performed using the experimental combustion chamber BKM, to investigate the wall side heat flux which is -- among other factors -- dependent on cooling film properties. To gain more insight into the film behaviour under real rocket-like conditions, optical diagnostics have been applied. The chosen methods were shadowgraphy and OH* imaging producing optical data sets which are analysed in this study. In this context, a description of the necessary background information is given, concerning rocket combustion chambers, film cooling and optical diagnostics of O2/H2 combustion. The applied methodology for optical analysis is described, followed by a presentation of the results. During the test campaign, it became clear that the optical setup was not optimised for creating meaningful shadowgraphy recordings which is why the shadowgraphy data has to be treated as flame emission imaging. The behaviour of the gas layer adjacent to the chamber wall could be characterised based on qualitative (luminosity, LOx shadow, reflection, recirculation zone and flame shape) and quantitative (layer thickness, layer length, pressure conditions) analysis. The thickness could be identified for each load step and an average length of the layer was found as well. OH* imaging has been used supplementary to support the observations from the flame emission images. An in depth frame by frame analysis was not possible due to time constraints. However, the time averaged images yielded results in accordance to the flame emission and could give a relative figure for the temperature distribution in the combustion volume. An artefact in the data was found, stemming presumably from the image intensifier. This artefact needs to be researched for a future error reduction in the data of this and other campaigns. Additionally, the thickness of the layer suggested a correlation to the models for film cooling efficiency. Such a correlation could not be established. Nevertheless, the film cooling models show the same behaviour as the data obtained from the flame emission imaging. Finally, suggestions are given how the data analysis and the optical setup could be improved for future, similar campaigns.