Nous étudions les mécanismes d’endommagement dus à l’érosion pluie sur des matériaux massifs et sur des matériaux revêtus. Pour cela, un banc d’essais spécifique a permis de mener des recherches en laboratoire. Le principe de l’essai repose sur l’émission de jets à haute vitesse et à haute fréquence.L’étude des matériaux massifs met en évidence un mécanisme de propagation de fissures par fatigue qui aboutit à des pertes de matière. Ces mêmes endommagements ont été observés sur des échantillons usés en service. Il a été confirmé que la dureté augmente la résistance à l’érosion pluie des matériaux métalliques.L’étude des revêtements s’est focalisée sur 2 technologies, à savoir la pulvérisation cathodique magnétron, qui est un procédé de déposition phase vapeur, et la projection thermique sous flamme supersonique. Les revêtements obtenus par projection thermique (dont l’épaisseur était supérieure à 200 μm), se sont révélés moins résistants à cause d’un manque d’adhérence ou de la présence de défauts au sein du matériau. Les revêtements obtenus par PVD (dont l’épaisseur était inférieure à 30 μm) ont permis d’obtenir des gains de résistance significatifs. Dans tous les cas, quel que soit la technologie utilisée, l’adhérence du revêtement s’est révélé être un paramètre critique en ce qui concerne la résistance de la surface à l’érosion pluie. Enfin, une simulation numérique en dynamique a permis d’étudier les champs de contraintes dans des feuillets métalliques, et ce, en faisant varier leurs épaisseurs, les matériaux qui les composent, et la vitesse d’impact. Les résultats de la simulation tendent à prouver que la propagation des ondes de contraintes dans le matériau peut entrainer des phénomènes de sur-contraintes dans les feuillets les plus fins à cause de la réflexion des ondes sur la face antérieure de la plaque. / The study deals with the rain erosion mechanisms of both bulk and coated materials. For that purpose, a specific test bench has been built, enabling laboratory research. The principle of the test is based on the emission of high-speed water jets at high frequency.Studying bulk materials confirmed the positive influence of hardness on rain erosion resistance for metallic materials. The mechanism found responsible for material losses is the propagation of fatigue cracks. These fatigue damages were also observed on in-service worn out samples.The analysis of coated materials focused on two covering technologies, namely physical vapor magnetron sputtering deposition (Magnetron PVD) and high velocity oxy-fuel coating spraying (HVOF). The coatings obtained by HVOF (> 200μm) weren’t resistant enough due to lack of adhesion and/or due to specific defects within the material. The coatings obtained by PVD (< 30μm) have yielded to significant improvements on the surface resistance. However, the adhesion of the coating appeared as a critical parameter for the rain erosion resistance.Finally, a numerical simulation has been designed to study dynamic stress fields in metal sheets. Moreover the model allowed the sheets thicknesses, the materials, and the speed of impact to vary. The simulation showed that the propagation of stress waves in the material may cause over-stresses phenomena in the thinner sheets, due to the reflection waves on the back side of the plate.

Dans le cadre de la modélisation et de l’expérimentation multi-échelles (projet LabEx MMCD pour les matériaux pour la construction durable) de l’Université Paris-Est Marne-la-Vallée, cette thèse de doctorat vise à modéliser et caractériser les micro-matériaux conçus par impact de gouttelettes de céramique fondue. Les applications de ces matériaux revétus de couches minces sont des traitements de surface pour la construction durable tels que la protection anti-corrosion, les barrières thermiques, le traitement du verre ou les renforts mécaniques. En particulier, nous nous concentrons sur la physique associée à la dynamique des gouttelettes liquides (l'aire de contact et le temps de contact entre la gouttelette et la surface) en effectuant une série de simulations numériques pour les écoulements diphasiques à petite échelle avec le code maison Thetis. Nous avons considéré des variations des conditions d'impact initiales ainsi que l’influence des forces d'inertie, capillaire et visqueuses sur la dynamique des gouttelettes. Nous nous sommes intéressés en particulier au diamètre d'étalement maximal, au temps d’étalement maximal et au temps de contact, sur des surfaces solides de mouillabilité variable. Le code est basé sur l’utilisation d’une méthode Volume-Of-Fluid. Il introduit une fonction auxiliaire régularisée pour estimer la courbure locale et la normale à l'interface. Les principaux liquides de référence adoptés sont l'eau et la céramique fondue, l'eau est choisie pour valider notre code en comparant les simulations aux résultats expérimentaux. La céramique fondue est adoptée car elle est largement utilisée en projection thermique pour créer des barrières thermiques et chimiques (couches anti-oxydantes) ainsi que des renforts mécaniques sur des échantillons spécifiques. Nous nous concentrons sur les cas où les surfaces sont hydrophobes, même si les cas hydrophiles sont également considérés dans les configurations de validation pour des raisons de généralité. Egalement, en introduisant une partie de calcul de l'énergie dans la thèse, une analyse énergétique détaillée de la gouttelette après l'impact est effectuée dans les phases d'étalement et de rétraction pour bien comprendre la dynamique à l'intérieur de la gouttelette. Nous trouvons que le temps de projection est inversement proportionnel à la vitesse d’impact, indépendamment de l’angle de contact lors de l’étalement au temps courts. Une nouvelle mise à l'échelle entre l'étalement maximal et le temps d'étalement est proposée. Celle-ci s'accorde très bien avec les résultats expérimentaux. Par ailleurs, nous introduisons cette mise à l’échelle dans une classe de modèle basée sur la conservation de l’énergie pour prédire l’étalement maximal adimensionné, ce qui permet de mieux prévoir l’étalement maximal adimensionné. Pour finir, une mise à l'échelle du temps de contact est proposée en termes de nombre d'Ohnesorge et de Reynolds / Under the framework of the LabEx Multi-Scale Modelling and Experimentation of Materials for Sustainable Construction, of Université Paris-Est Marne-La-Vallée, the present PhD thesis aims at modelling and characterizing micro-material designed by impact of molten ceramic droplets. The applications of thin coating materials are surface treatments for sustainable construction such as anti-corrosion, heat barrier, glass treatment or mechanical reinforcement of specific structures.In particular, we focus on the physics behind the liquid droplets' dynamics (the contact area and the contact time between the droplet and surface) by conducting a series of small scale multiphase flow numerical simulations with home-made code Thetis. All simulations are axisymmetric. We have considered variations of initial impact conditions, and studied the influence of inertial, capillary and viscous forces on the droplets' dynamics, especially the maximum spreading diameter, spreading time and the contact time, on solid surfaces. The code is based on Volume-Of-Fluid techniques and introduces an auxiliary smooth function to estimate the local curvature and the normal to the interface. The major reference liquid adopted are the water and the molten ceramic, the water is chosen to validate our code against available experiments at the beginning. The molten ceramic is adopted as it is widely used in thermal spray to built thermal and chemical barriers (anti-oxidant layers) as well as mechanical reinforcements on specific samples. We focus on the cases in which the surfaces are hydrophobic, even if hydrophilic cases are also considered in validation configurations for the sake of generality. Meanwhile, by introducing an energy calculation part in the code, a detailed energetic analysis of the droplet after impact is performed in both the spreading and retraction stage to have a deep understanding of the dynamics inside the droplet.We find the jetting time is inversely proportional to the impact velocity, independent of the contact angle in the early spreading. A new scaling between maximum spreading and spreading time is observed, and agrees well with experimental results. Further, we introduce this scaling into the model based on energy conservation to predict the maximum spreading factor, which provides better prediction on maximum spreading factor than existing literature references. Also a scaling of contact time is proposed in terms of Ohnesorge number and Reynolds number

This dissertation explores the physics of droplet impingement on superhydrophobic surfaces. The research is divided in three categories. First, the effect of a slip boundary condition on droplet spreading/retracting is considered. A model is developed based on energy conservation to evaluate spreading rates on surfaces exhibiting isotropic and anisotropic slip. The results show that larger slip causes the droplet to spread out farther owing to reduced friction at the interface for both slip scenarios. Furthermore, effects of slip become magnified for large Weber numbers due to the larger solid-liquid contact area during the process. On surfaces with anisotropic slip, droplets adopt an elliptical shape following the azimuthal contour of the slip on the surface. It is common for liquid to penetrate into the cavities at the superhydrophobic interface following droplet impact. Once penetrated, the flow is said to be in the Wenzel state and many superhydrophobic advantages, such as self-cleaning and drag-reduction, become negated. Transition from the Wenzel to the Cassie state (liquid resides above the texture) is referred to as dewetting and is the focus of the second piece of this dissertation. Micro-pillar pitch, height and temperature play a role on dewetting dynamics. The results show that dewetting rates increase with increasing pillar height and increasing surface temperature. A scaling model is constructed to obtain an explanation for the experimental observations and suggests that increasing pillar height increasing the driving dewetting force, while increasing surface temperature decreases dissipation. The last piece of work of this dissertation entails droplet impingement on superheated surfaces (100°C - 400°C). We find that the Leidenfrost point (LFP) occurs at a lower temperature on a hydrophobic surface than a hydrophilic one, where the LFP refers to the lowest temperature at which secondary atomization ceases to occur. This behavior is attributed to the manner in which vapor bubbles grow at the solid-liquid interface. Also in this work, high-speed photographs reveal that secondary atomization can be significantly suppressed on a superhydrophobic surface owing to the micro-pillar forest which allows vapor to escape hence minimizing bubble formation within the droplet. However, a more in-depth study into different superhydrophobic texture patterns later reveals that atomization intensity can significantly increase for small pitch values given the obstruction to vapor flow presented by the increased frequency of the pillars.

In this thesis investigations are carried out on two research topics in context to spray drying. The first research topic is the production of dried particles having uniform characteristics. The second research topic is the development of new applications of the reaction engineering approach which, in recent times, has emerged as an effective tool to formulate drying kinetics models. The reaction engineering approach is also implemented to simulate the drying of monodisperse droplets corresponding to the experimental work in the first research topic. Manufacturing micron- and nano-sized particles having uniform characteristics has recently become a popular research area due to the unique functionalities of these kinds of particles in biomedical, drug delivery, functional foods, nutraceuticals, cosmetics and other valuable applications. Spray drying has been a common and economical route to produce dried particles. A typical characteristic of spray dried products is the existence of a significant variation in particle properties such as size and morphology. One possible idea to restrict this product non-uniformity is to achieve a good control over the droplet’s behaviour and characteristics inside the drying chamber. The current thesis has investigated an innovative spray drying technique, i.e. a single stream drying approach in order to restrict product non-uniformity. In this drying approach, identical sized droplets having vertical trajectories are dried under controlled gas flow conditions. The piezoelectricity-driven monodisperse droplet generator is used as the atomizer to disperse liquid droplets. A prototype single stream dryer was assembled based on the single stream drying approach using various components designed in the laboratory and several parts purchased from the market. Experiments were carried out using aqueous lactose solutions as a model system in order to check the practicability of manufacturing uniform-sized spherical particles. Preliminary results were found to be positive and reported in this thesis. Mathematical models on the drying of monodisperse droplets were developed in order to predict important droplet and gas parameter profiles during single stream drying. These models serve as a platform for design, optimization and scale-up purposes. Several important advantages and drawbacks of single stream drying are also reported. Problems encountered during the experimental work and future recommendations are presented in detail so that a more robust and effective drying research tool can be developed in future. Recently the reaction engineering approach (REA) has emerged as a simple and reliable technique to characterize the drying of various food and dairy materials. In this thesis two new applications of the REA are described for the first time in context to convective drying of aqueous droplets. The REA is used in this study to formulate the drying kinetics model for the drying of aqueous sucrose and maltodextrin (DE6) droplets. The effect of initial moisture content was explicitly demonstrated. The development of a new ‘composite’ REA which aimed to model the drying of aqueous droplets containing multiple solutes has been described. The composite REA was found to be suitable to characterize the drying behaviour of aqueous sucrose-maltodextrin mixtures of different proportions. The second new application of the REA is the development of a procedure to estimate surface properties of aqueous droplets during drying. In literature various droplet characteristics such as surface moisture contents were normally estimated using the diffusion-based drying kinetics model or the receding interface model. Surface moisture content and surface glass transition temperature profiles were evaluated here using a lumped-parameter model (REA) during the drying of aqueous sucrose, maltodextrin (DE6) and their mixtures. The same experimental data used for the development of the composite REA were used to yield predictions. The procedure was found to be useful in estimating surface moisture contents and understanding the stickiness behaviour of sugar droplets during drying. During the formulation of the REA-based drying kinetics model in this thesis, the assumption of uniform temperature within droplets was used. In most studies published in literature the uniform temperature assumption was justified by calculating the heat-transfer Biot numbers at the beginning and end of drying. However, the conventional Biot number concept does not take into account the evaporation effect and therefore would not be suitable to drying scenarios. In this thesis, a new approximation procedure is developed to estimate surface-centre temperature differences within materials following the entire drying process. This new procedure was helpful to check the extent of temperature non-uniformity within skim milk droplets under isothermal laboratory conditions as well as industrial spray drying conditions. Both conventional and drying-based Biot numbers are calculated and compared. Predictions showed that temperature gradients within the droplets were negligible during the drying of suspended droplets under laboratory drying conditions (slow drying), whilst the gradients were small and existed only for a short drying period for small droplets under industrial spray drying conditions (fast drying). Furthermore, it was observed that the maximum temperature gradient within the droplets did not exist at the starting or end points of the drying process, and therefore the estimation of Biot numbers at the starting and end point does not reflect temperature non-uniformity under drying conditions. This is a significant theoretical development in the area.

This thesis examines the particle capture, fibre wetting and droplet flow processes within wet filters collecting solid and liquid aerosols and within filters collecting only liquid aerosols. The processes involved in this type of filtration were examined through a series of experiments and models developed to describe the behaviour of fibre/liquid systems. This work can be summarized in 4 categories: (1) The bounce and immediate re-entrainment of liquid and solid monodisperse aerosols under a stable filtration regime (pre cake formation) by wet and dry fibrous filters. In this work it was found that the solid particles generally exhibited a lower fractional filtration efficiency than liquid particles (of the same size), although this difference decreased in the smaller size fractions. However, for the wet filtration regime (each fibre of the filter was coated by a film of water), no significant difference in filtration efficiency was detectable between solid and liquid aerosols. Either the bounce effect of the particles is inhibited by the liquid film, or the filtration conditions in the wet filter are so different that the aerosol properties are less significant with respect to capture. (2) A microscopic study of the effect of fibre orientation on the fibre wetting process and flow of liquid droplets along filter fibres when subjected to airflow and gravity forces was conducted. The flow of the liquid collected by the fibres was observed and measured using a specially developed micro-cell, detailed in the thesis. The experimental results were compared to a theoretical model developed to describe the flow of droplets on fibres. The theory and experimental results showed a good agreement. A sensitivity analysis of the model was performed which showed the droplet radius to be the most significant parameter. The model has the potential to improve filter self-cleaning and minimise water use. (3) An experimental study of the capture of solid and liquid (oil) aerosols on fibrous filters wetted with water. Variable quantities of liquid irrigation were used, and the possibility for subsequent fibre regeneration after clogging or drying was also studied. It was found that self-cleaning (removal of solid aerosols by water) occurred even under heavily dust-laden conditions, and post evaporation of water. With the collection of oil aerosols on fibres wetted with water, a predominance of the barrel shaped droplet on the fibre was observed, with oil droplets displacing water droplets (if the oil and fibre combination created a barrel shaped droplet), creating various compound droplets of oil and water not previously reported in literature. (4) An extensive experimental investigation of the wetting processes of fibre/liquid systems during air filtration (when drag and gravitational forces are acting) has shown many important features, including droplet extension, oscillatory motion, and detachment from fibres as airflow velocity increases. The droplet oscillation is believed to be induced by the onset of the transition from laminar to turbulent flow as droplet size increases. To model such oscillation it was necessary to create a new conceptual model to account for the forces both inducing and preventing such oscillation. The agreement between the model and experimental results is satisfactory for both the radial and transverse oscillations.

La conservation des ressources génétiques végétales est un défi majeur, le maintien de la biodiversité étant capital pour répondre aux besoins des générations futures. Ce travail de thèse, s'inscrivant dans cette démarche de conservation, a été réalisé sur Pelargonium, plante ornementale multipliée végétativement. Un procédé de cryoconservation d'apex par « dropletvitrification » a été mis en place avec succès sur P. x peltatum ‘Balcon Lilas'. Avec un taux de régénération moyen de 40 % obtenu sur 28 taxons testés, la constitution d'une cryobanque est maintenant possible. L'optimisation de la survie post-cryoconservation passant par une meilleure connaissance des effets des solutions cryoprotectrices sur les cellules, des études microscopiques faisant appel notamment à la RTM (Real Time Microscopy) et la CLSM (Confocal Laser Scanning Microscopy) ont été pratiquées. Elles ont permis de mettre en évidence, d'une part que la solution de vitrification (PVS2) provoque une contraction importante et rapide des cellules en agissant sur les parois et d'autre part que la solution de charge (LS), outre son rôle d'osmoticum, atténue les altérations dues à PVS2. Les travaux sur la régénération in vitro à partir d'apex ont montré que celle-ci était le plus souvent directe ce qui peut favoriser le maintien de la conformité. Ainsi, une plante obtenue après régénération d'un apex cryoconservé ou non, ne présente pas de modifications phénotypiques sauf chez certains cultivars chimériques. Enfin, des tests ELISA ont montré une augmentation du taux d'assainissement lors d'une régénération après cryoconservation par rapport à une simple culture d'apex pour deux virus (PFBV et PLPV). Cependant les immunolocalisations réalisées ont mis en évidence que les virus étaient toujours présents dans les apex, l'élimination n'étant que partielle. L'augmentation de ces connaissances devrait permettre une utilisation à plus grande échelle de la cryoconservation participant ainsi à une meilleure gestion de la conservation des ressources génétiques par une offre stratégique plus importante.

A new method is proposed for correctly modeling the long range interaction between a fluid and a bounding wall in atomistic simulations. This method incorporates the molecular structure of the solid substrate while allowing for a finite interaction cutoff by making a proper estimation of long range correction for the fluid-wall interaction. The method is then applied to a molecular dynamic simulation of a spreading droplet. Conparison to simulations using several other previously used methods shows that the long range correction can be significant in some circumstances. / Singapore-MIT Alliance (SMA)

In this thesis, the application of electrospray ionisation mass spectrometry (ESI-MS) to the analysis of intact proteins is demonstrated. In papers I and II, the use of ESI-MS for the analysis of noncovalent protein-ligand complexes were discussed. In addition, the interfacing of liquid chromatography (LC) with ESI-MS and the development of an LC-ESI-MS method were demonstrated in paper III for the quality control of recombinant proteins. Furthermore, this method was applied in paper IV for the analysis of covalent glycosyl-enzyme intermediates. The monitoring of noncovalent complexes by ESI-MS is well established. However, the varying characteristic of ESI-MS data, especially in the analysis of noncovalent complexes can make the quantification of such complexes troublesome. In paper I, it was demonstrated how the variation in the position of the ESI-emitter and the initial droplet size of the electrosprayed droplets, together with different partitioning of a protein and its ligand in these droplets, can be the cause of such varying characteristics. Furthermore, it was shown that the partitioning can be of electrostatic and/or hydrophobic/hydrophilic origin. Thus it was demonstrated that if the ligand is more hydrophobic and thereby more surface active relative to the protein, decreasing the droplet size or increasing the distance between the electrospray emitter and the sampling orifice will lead to more efficient sampling of the droplet bulk where the ligand concentration is low. This results in a favoured sampling of free protein relative to the protein ligand complex. The opposite was shown to occur if the ligand is more hydrophilic than the protein. In paper II, Ribonuclease A (RNAse) was used as a model for enzymes acting on polymeric substrates with different chain lengths. Nano-ESI-MS was applied to monitor the noncovalent interactions between RNAse and different target ligands. Among the single building blocks of RNA, including ribose, the bases adenine, guanine, cytosine and uracil, and phosphate, only phosphate was observed to interact at multiple RNAse sites at a higher cone voltage. Furthermore, monobasic singlestranded deoxycytidylic acid oligomers (dCx) of different lengths (X=6, 9 and 12), and RNAse were analysed with nano-ESI-MS. The deoxycytidylic acid with 12 nucleotides was observed with the highest complex to free protein ratio, hence indicating the strongest interaction. Finally, collision induced dissociation of the noncovalent RNAseA-dC6 complex resulted in dissociation of covalently bound cytosine from the nucleotide backbone rather than break up of the noncovalent complex, illustrating the cooperative effect of multiple noncovalent interactions. In paper III an LC-ESI-MS method was presented capable of analysing proteins 10-100 kDa in size, from salt-containing liquid samples. The proteins included human protein fragments for the largescale production of antibodies and human protein targets for structural determination, expressed in E. coli. Also, glycosylated proteins expressed in Pichia pastoris were analysed. The method provides fast chromatography, is robust and makes use of cheap desalting/trap columns. In addition it was used with optimised reduction and alkylation protocols in order to minimize protein aggregation of denatured and incorrectly folded proteins containing cysteins, which otherwise form adducts by disulfide bond formation. Furthermore, the method was used in paper IV for the quantification of covalent proteinligand intermediates formed enzymatically between PttXET16-34, a xyloglucan endo-transglycosylase (XET) from hybrid aspen, and the synthetic substrates GalGXXXGGG and GalXXXGXXXG designed in order to function as donor substrates only. Thus covalent GalG-enzyme and GalGXXXG-enzyme complexes were detected. Moreover, establishing of a pseudo equilibrium for the formation of the covalent GalGXXXG-enzyme complex enabled quantification of the saccharide and enzyme constituents of this equilibrium and determination of the free energy of formation (∆G0). The high mass resolution of the TOF-MS allowed unambiguous assessment of the covalent nature of the glycosyl-enzyme complexes. Morover, the formation of noncovalent complexes between excess substrate and protein, which can deteriorate MS-signal and increase spectrum complexity, was efficiently avoided by the chromatographic step, which separated the saccharide content from the protein content. / QC 20100913

Hepatic lipase (HL) plays a pivotal role in the catabolism of apolipoprotein (apo)B-containing lipoproteins and high density lipoprotein (HDL) particles through its reported catalytic and non-catalytic extracellular functions. The current study tested the hypothesis that HL expression might impair formation and secretion of hepatic derived very low density lipoproteins (VLDL) and apoA-I (nascent HDL). Stable or transient expression of human HL (hHL) in McA-RH7777 cells resulted in decreased incorporation of [3H]glycerol into cell-associated and secreted (VLDL-associated) 3H-triacylglcyerol (TAG) relative to control cells. Stable expression of catalytically-inactive hHL (hHLSG) also resulted in decreased secretion of VLDL-associated 3H-TAG whereas cell-associated 3H-TAG levels were unchanged. Expression of hHL or hHLSG increased cell-associated 35S-apoB100 with relatively no change in secreted 35S-apoB100. Importantly, hHL or hHLSG expression resulted in reduced 3H-TAG associated with the microsomal lumen lipid droplets (LLD), and increased relative expression of ApoB and genes involved in lipogenesis and fatty acyl oxidation. Transient expression of hHL in HL-null primary hepatocytes, mediated by adenoviral gene transfer, resulted in decreased steady-state levels of cell-associated and secreted apoA-I and reduced rates of synthesis and secretion of 35S-apoA-I. HL-null hepatocytes exhibited increased levels of secreted 35S-apoA-I relative to wildtype hepatocytes while cell-associated 35S-apoA-I levels were normal. Transient expression of a hHL chimera (hHLmt), in which the C-terminus of hHL was replaced with mouse HL sequences, exerted an inhibitory effect on apoA-I production similar to that of hHL even though hHLmt was secreted less effectively than hHL with impaired exit from the endoplasmic reticulum (ER) as compared with hHL. In contrast, stable expression of hHL in McA-RH7777 cells resulted in a dose-dependent increase in cell-associated and secreted 35S-apoA-I levels. These studies demonstrate that hHL has an intracellular (but non-catalytic) role in reducing the content of the LLD and ultimately the buoyancy of secreted VLDL particles, and that the N-terminal sequences of ER-residing hHL directly or indirectly modulates the production and secretion of apoA-I (nascent HDL) from hepatocytes.

Hepatic lipase (HL) plays a pivotal role in the catabolism of apolipoprotein (apo)B-containing lipoproteins and high density lipoprotein (HDL) particles through its reported catalytic and non-catalytic extracellular functions. The current study tested the hypothesis that HL expression might impair formation and secretion of hepatic derived very low density lipoproteins (VLDL) and apoA-I (nascent HDL). Stable or transient expression of human HL (hHL) in McA-RH7777 cells resulted in decreased incorporation of [3H]glycerol into cell-associated and secreted (VLDL-associated) 3H-triacylglcyerol (TAG) relative to control cells. Stable expression of catalytically-inactive hHL (hHLSG) also resulted in decreased secretion of VLDL-associated 3H-TAG whereas cell-associated 3H-TAG levels were unchanged. Expression of hHL or hHLSG increased cell-associated 35S-apoB100 with relatively no change in secreted 35S-apoB100. Importantly, hHL or hHLSG expression resulted in reduced 3H-TAG associated with the microsomal lumen lipid droplets (LLD), and increased relative expression of ApoB and genes involved in lipogenesis and fatty acyl oxidation. Transient expression of hHL in HL-null primary hepatocytes, mediated by adenoviral gene transfer, resulted in decreased steady-state levels of cell-associated and secreted apoA-I and reduced rates of synthesis and secretion of 35S-apoA-I. HL-null hepatocytes exhibited increased levels of secreted 35S-apoA-I relative to wildtype hepatocytes while cell-associated 35S-apoA-I levels were normal. Transient expression of a hHL chimera (hHLmt), in which the C-terminus of hHL was replaced with mouse HL sequences, exerted an inhibitory effect on apoA-I production similar to that of hHL even though hHLmt was secreted less effectively than hHL with impaired exit from the endoplasmic reticulum (ER) as compared with hHL. In contrast, stable expression of hHL in McA-RH7777 cells resulted in a dose-dependent increase in cell-associated and secreted 35S-apoA-I levels. These studies demonstrate that hHL has an intracellular (but non-catalytic) role in reducing the content of the LLD and ultimately the buoyancy of secreted VLDL particles, and that the N-terminal sequences of ER-residing hHL directly or indirectly modulates the production and secretion of apoA-I (nascent HDL) from hepatocytes.