Lipid Classification, Functions, Occurrence in Foods, Normal & Abnormal Digestion & Absorption

Overview - Classification, structure and nomenclature of lipids Fatty acids and their derivatives - Physical, chemical and functional properties of saturated fatty acids, monounsaturated fatty acids, and the omega-3, -6 and -9 families of polyunsaturated fatty acids; types and content in specific foods; genetic or chemical alteration of food fatty acids; effects of processing, storage and cooking practices on fatty acids in foods Acylglycerides and derivatives - mono-, di- and triacylglycerols; phosphoglycerides; Glycolipids Sphingolipids Sterols and other isoprenoid compounds Tocopherols, tocotrienols, polyphenols, and other fat-soluble food components, vitamins and xenobiotics Dietary recommendations for fats - Estimation of dietary fat intake; nutritional assessment of fats; Labeling and analytical methods for food lipids; Role of fats and fat replacers in medical or designer foods; Review of dietary fat in various populations, human health and etiology of chronic diseases Digestion and absorption of nutrient lipids and other fat-soluble compounds - Gastric, intestinal and pancreatic enzymes; Parasympathetic and sympathetic influences; Endocrine and paracrine influences; Cholagogues, hepatic bile, gallbladder bile and enterohepatic circulation; Physical aspects of lipid digestion (solubilization by bile salts and other bile lipids, mechanism of action of lipase, colipase and bile acids, diffusion through the unstirred water layer); Absorption into and packaging within the enterocyte (differences in fates of long vs. medium chain fatty acids, role of enterocyte fatty acid binding proteins, importance of fatty acid chain length in infants and in malabsorption disorders) Disorders of lipid digestion and absorption - Deficiencies of lipase, colipase or bile; Sprue, abetalipoproteinemia and other malabsorptive syndromes; Special considerations for neonates and premature infants (reasons for impaired fat utilization, problems associated with prolonged enteric fasting)

Pathways of Fatty Acid Synthesis

Relationships of dietary and endogenous fatty acids - Essential fatty acids; Monoene/diene/ polyene ratios Transport of fatty acids - Transport by plasma albumin; Transport into peripheral cells; Transport across the placenta; Transport of fatty acids from intestine and liver and from adipose Fatty acid biosynthesis in liver, adipose and mammary gland - General effects of diet on fatty acid biosynthesis (amount of dietary fat and other nutrients, response to high carbohydrate diets and GLUT-4 over-expression); Response of lipogenic enzymes to dietary SF, MUF, PUF; Response to trans fatty acids; Influence of dietary zinc and other minerals on lipogenic enzymes; Movement of acetyl CoA into cytosol (tricarboxylate transport system, ATPcitrate lyase, malate dehydrogenase and malic enzyme); Acyl carrier protein (ACP) Acetyl CoA carboxylase (ACC): Regulation of expression and activity; Specific long term dietary effects; Effects of glucagon and insulin (cAMP-mediated phosphorylation/dephosphorylation, general effects on gene expression); Differential splicing of promotor primary transcripts; Differences in sensitivity of ACC promotors I (ACCP-I) and II to dietary fat; Ribozyme gene regulation; Allosteric modulation by citrate and long chain acyl CoA; ACCP-I in obese Zucker (fa/fa) rat animal model Fatty acid synthetase (FAS) - Unique structure of enzyme and relationship to mechanism (acyl carrier protein, required cofactors); Dietary and hormonal influences on the FAS promotor and on FAS gene expression; Interactions of effects of insulin, glucagon, somatotropin and thyroxin; Glucose regulation of FAS mRNA stability; Role of S14 protein; Allosteric activation by phosphorylated sugars; Tissue specific differences; Inhibition of synthetase; FAS in brown adipose tissue (BAT); Production of branched chain fatty acids from methylmalonyl CoA Elongation and retroconversion - Cell specific differences in elongases; Importance, use and abundance of elongation products; Control of reactions in mitochondria vs. endoplasmic reticulum; Very long chain fatty acid synthesis in nervous tissues; Retroconversion enzymes (conversion of adrenic acid to arachidonic acid, importance and use of chain-shortened fatty acids, influence of diet on control of enzymes Desaturation - General mechanism of the desaturases and electron transport system; -9 desaturases (products, control of gene expression, regulation of enzyme activity); -6 Desaturases (synthesis of arachidonic from linoleic acid, other products, inhibition by polyunsaturated fatty acids and trans fatty acids); -5 Desaturases (products, stimulation and inhibition of activity, desaturation of elaidic acid); -4 Desaturases (products, tissue specific distribution);NADPHlinked reduction of acyl CoA to fatty alcohol Disorders of fatty acid synthesis and transport - Essential fatty acid deficiency (in pregnancy and early human development, in patients with chronic gastrointestinal disorders, in patients on long term parenteral nutrition, association with down regulation of protein kinase C-); Total parenteral nutrition-induced lipogenesis in critically ill patients; Biotin deficiency and acetyl CoA carboxylase; Defects in the regulation of FAS (model: Genetically obese JCR:LA-corpulent (cp) rats, adipsin effects, hyper- and hypothyroidism); Analbuminemia

Triglyceride, Phospholipid and Sphingolipid Synthesis

Common Pathway - Synthesis of phosphatidic acid (regulation of glycerol-3-phosphate acyltransferase, regulation of dihydroxyacetone-phosphate acyltransferase); Biochemical functions of phosphatidic acid (PA) and diacylglycerol (DAG): Branch point in pathway (regulation of flux between PA phosphohydrolase and CDP-DAG synthetase, distribution and regulation of enzymes) Synthesis of triglyceride in hepatocytes and adipose cells - Determination of flux through esterification pathway (importance of fatty acid and CDP-choline supply, activity of DAG:acyltransferase, ATP-dependent inactivation of adipose DAG:acyl-transferase, importance of dihydroxyacetone-phosphate to triglyceride synthesis in adipose); Effect of diet on regulation of major enzymes in pathway from glycerol-phosphate; Use of permeabilized rat hepatocytes as a cell model Phospholipid synthesis - Formation of phosphatidylinositol (characterization and regulation of phosphatidyl inositol (PI) synthase, regulation of the PI 4-kinase, PI 3-kinase and PI 5-kinase families, importance of phosphorylations, activation of enzymes by various growth factors); Importance as precursor of signal molecules; Importance in structure of cell membranes Synthesis of phosphatidylserine (PS) and phosphatidyl ethanolamine (PE) - Base exchange; Characteristics and regulation of PS synthase; PS decarboxylase; Ethanolamine/choline kinase and CTP: phospho-ethanolamine cytidyltransferase; Importance of the CDP-ethanolamine pathway; Age-related changes in PE levels in certain cells Synthesis of phosphatidylcholine (PC) Importance of supply of CDP-choline; Regulation of choline kinas; CTP:phosphocholine cytidyltransferase and DAG:cholinephosphotransferase; Distribution and role of PE methyltransferases (PEMT); Coordinate control of gene expression with CTP:phosphocholine cytidyl-transferase; Use of McArdle R147777 rat hepatoma model for deficiency of enzyme; Properties of dipalmitoyl phosphatidylcholine Synthesis of cardiolipin - Abundance and importance; Regulation of synthesis Synthesis of ether phospholipids - Pathways from dihydroxyacetone phosphate; Platelet activating factor (unique properties and function, regulation of its synthesis, relationship to other phospholipids); Plasmalogens (desaturase action, regulation and importance) Phospholipid activating protein (PLAP) Function; Activation by PKC-MAP kinase cascade Coordinate regulation of phospholipid synthesis - Cis regulatory element for inositol-choline mediated regulation of phospholipid synthesis; Role of PKC; Regulation in conjunction with FAS genes; Importance of fatty acid binding protein Sphingolipid synthesis - Synthesis of ceramide from sphingosine and behenic acid or other long chain fatty acids; 3ketosphinganine synthase and reductase; Acyltransferase and dihydroceramide reductase; Formation of ceramide with sphingomyelinase (role of ceramide in the TNF-signaling pathway, effect of ceramide on expression of phospholipase A2 and cyclooxygenase); Sphingomyelin synthesis from lecithin and ceramide; Sphingomyelin synthetase (activation by translocation, use of choline-deficient cultured cell models); Tissue specificity Sphingoglycolipids - Galacto- and glucocerebrosides(role of UDP, abundance and induction of enzymes); Sulfatides (galactocerebroside-3-sulfate)(abundance in the brain, role of 3'-phosphoadenosine-5'-phospho-sulfate); Gangliosides [acidic ceramide oligosaccharides] and globosides [neutral ceramide oligosaccharides] (formation of precursor lactosylceramide, differences in Gm, GD, GT and GQ ganglioside series, heterogeneity in expression of ganglioside synthase genes, differences in synthetic rates between differentiating and mature oligodendroglia) Sorting of sphingolipids to cellular domains Disorders - Cell mutants defective in synthesis of glucosyl-phosphatidyl inositol; Anticardiolipin anti-bodies; Primary antiphospholipid syndrome; Deficiency of sulfatide synthesis in polycystic (cpk/cpk) mice

Isoprenoid and Sterol Synthesis

Overview Importance and significance of isoprenoid compounds - Isopentenyl-tRNA; Ubiquinone, dolichol and other polyprenol compounds; Prenylation of proteins; Steroid compounds General pathway for isoprenoid synthesis - Locations of enzymes; General regulation of synthetic pathway HMG-CoA synthetase [3-Hydroxy-3-methylglutaryl CoA:Acetoacetyl CoA lyase] - Chemistry of catalysis; Homology to mitochondrial synthetase; Regulation of enzyme (sterol-mediated regulation of transcription, Sterol regulatory element-binding protein-1 (SREBP-1), proteolysis of transcription factor , effect of oxysterols, negative transcription factors, effect of triiodothyronine, phosphorylation effects, pharmacologic inhibition to reduce cholesterol synthesis); Characterization of the recombinant and mutant enzymes (cloning and expression studies in E. coli, differences in genes for cytosolic and mitochondrial synthase enzymes) HMG-CoA reductase [Mevalonate:NADP+ oxidoreductase] - Location, structure and characteristics (glycosylation of enzyme, hydrophobic regions in membrane of endoplasmic reticulum, cytosolic catalytic site, effect of phosphorylation on catalytic activity and half-life of the enzyme); Chemistry of catalysis; Regulation of enzyme by covalent modification (reductase kinase and reductase kinase kinase, phospho-protein phosphatase-1, phosphoprotein phosphatase-1 inhibitor, effects of hormones on state of phosphorylation, reductase activating protein); Regulation by multivalent, allosteric control (sterols and oxysterols, excess mevalonate, other nonsterol metabolites of pathway);

Transcriptional vs. post-translational regulation (diurnal variation, sterol regulatory element; homology with that of HMG CoA synthetase and the LDL receptor, effects of phorbol esters); Species differences in enzyme regulation; Differences in activity among organs and tissues; Differences in activity in various physiologic states (growth factors, age, obesity, gender, diabetes mellitus, methods for studying amount and activity); Pharmacologic inhibition of reductase (types and actions of inhibitors, effectiveness in suppressing cholesterol synthesis, consequences of inhibition on protein prenylation, concerns about use of reductase inhibitors in pregnancy and in children); Effects of other compounds or conditions on reductase activity (Cu and glutathione, propionic and butyric acids, long chain fatty acids, fasting or increased dietary carbohydrate or dietary fat, bile acids Farnesyl diphosphate synthase - Characterization of enzyme; Molecular cloning studies Squalene synthase - Characteristics and locations of enzyme; Importance as branch point in pathway; Regulation of activity (coordinate regulation with reductase, effects of reductase competitive inhibitors, allosteric effectors) Sterol carrier protein Squalene expoxide cyclase - Characteristics of enzyme and chemistry of reaction; Regulation Additional steps in conversion of lanosterol to cholesterol Important facets of cholesterol homeostasis - Functions and fates of cholesterol (significance of the cholesterol: phospholipid ratio in membranes, synthesis of vitamin D, bile acids and steroids); Importance of 7--hydroxylase, LCAT and ACAT to cholesterol metabolism; Effects of dietary isoprenoid and steroid compounds on cholesterol synthesis (limonene and other monoterpenes, -sitosterol and other phytosterols, other polyprenyl compounds); Effects of phenobarbital and other drugs which induce the cytochrome P450 system; Exogenous vs endogenous cholesterol Methods of assessing cholesterol metabolism - Sterol balance studies in humans and animals; Perfused organ studies; Tissue culture studies; Enzyme assays on subcellular fractions; Labeled precursor studies; Breath isoprene excretion Disorders - Smith-Lemli-Opitz syndrome; Homozygous and heterozygous familial hypercholesterolemia

Lipid Modifications of Proteins

Overview Prenylated proteins- Functions of prenylation; Prenyl transferases and specificity (farnesylated residues, geranylgeranylated residues, sites of modification, mechanism of reactions); Prenyl-dependent targeting of receptor kinases; Metabolism of prenylated proteins; Regulation of prenylation (effect of inhibition of HMG CoA reductase, inhibition of transferases, implications of regulation) Acylated Proteins - Functions and locations of acylated proteins; Types of acylation (myristoylation, Ca++-myristate switch protein, myristate and ADP-ribosylation factors, myristoyl CoA: Protein-N-myristoyl transferase, palmitoylation, ras acylation sites and G-protein-coupled receptors); Protein sites of modification (cotranslational modification, post-translational modification); Differences in turnover rates of myristic and palmitic modifications; Regulation of myristoyl CoA pools Glycosylphosphatidylinositol (GPI) modification - Types of proteins modified; Core tetrasaccharide; Release of polypeptide by specific phospholipase Disorders of modification

Oxidation of Fatty Acids

Overview Acyl CoA synthetases (thiokinases) Carnitine palmitoyl transferases I and II and other transferases - Transport across mitochondrial membrane; Regulation of enzymes -Oxidation - Location and characterization of mitochondrial enzymes; Acyl CoA dehydrogenases (Electron-transfer flavoprotein (ETF), ETF: ubiquinone oxidoreductase); Enoyl CoA hydratase; 3-Hydroxyacyl CoA dehydrogenase; Ketoacyl CoA thiolase Peroxisomal -oxidation - Importance to long chain fatty acid metabolism; Reaction sequence (acyl CoA oxidase, hHydratase and hydroxyacyl CoA dehydrogenase, thiolase); Peroxisomal proliferation (induction by hypolipidemic drugs, homology of peroxisomal proliferation activation receptor with the steroid hormone receptor superfamily, animal and cell models for studying regulation of proliferation in normal and tumor cells, importance in fatty acid metabolism) Oxidation of unsaturated fatty acids - Substrate specificity; Mechanism of enoyl CoA isomerase; 2,4-Dienoyl CoA reductase; 3,2-Enoyl CoA isomerase Oxidation of odd chain fatty acids - Propionyl CoA carboxylase and methylmalonyl CoA mutase; Roles of biotin and vitamin B12 Other oxidation pathways - -Oxidation of branched chain fatty acids; -Oxidation Ketone body metabolism - Normal ketogenesis in liver mitochondria (acetyl CoA acetyltransferase (thiolase), hydroxy--methylglutaryl (HMG) CoA synthase, HMG CoA lyase); Utilization in muscles and heart (3-ketoacyl CoA transferase, importance of pathway); Regulation during fasting; Lack of insulin

Disorders of fatty acid and ketone body oxidation - Deficiencies of carnitine or carnitine palmitoyl transferase (generalized systemic carnitine deficiency due to genetic defect in carnitine synthesis or renal reabsorption defect, myopathic carnitine deficiency, secondary carnitine deficiency in organic acidurias, genetic defect in carnitine palmitoyl transferase, rationale for diet therapy); Long chain acyl CoA dehydrogenase deficiency [LCAD] (most frequent site of mutation, therapeutic intervention and rationale); Medium chain acyl CoA dehydrogenase deficiency [MCAD] (most frequent site of mutation, occurrence in sudden infant death syndrome, therapeutic intervention and rationale); Short chain dehydrogenase deficiency (SCAD); Jamaican vomiting sickness; X-adrenoleukodystrophy (XALD) due to deficiency of peroxisomal acyl CoA synthase; Rhizomelic chondrodysplasia punctata; Refsum's disease, a defect in -oxidation; Zwelleger's syndrome; Diseases of propionate and methylmalonate metabolism (vitamin B 12 deficiency, methylmalonyl CoA mutase defects, propionyl CoA carboxylase deficiency); Ketoacidosis in untreated diabetes mellitus Manipulation of type and amounts of fatty acids during therapeutic intervention - Types of lipid emulsions; Physiologic acute effects of the various fatty acids in the emulsion; Usefulness in treatment of specific disease states

Degradative Pathways for Acylglycerides and Sphingolipids

Overview - ubiquitous enzymes with certain functional and enzymatic attributes in common. Common ancestral gene for glycerolipid hydrolases (lipases) -.Homologies (ex. pancreatic TG lipase,, hepatic TG lipase and LPL have about 30% homology of aa sequence); General mechanism of action (Interfacial vs. catalytic site binding, role of cofactor or activator proteins, kinetic properties, lipid transfer activity) Digestive lipases - Occurrences, structures, features; Lingual lipase; Pregastric lipase; Gastric lipase; Pancreatic lipases;. Colipase; Pancreatic lipase-related proteins 1 and 2 (PLRP1 & 2); Crystallographic structures; Attributes (substrate specificity, interfacial activation, bile salt inhibition and colipase reactivation, mechanism of catalytic triad, site-specific mutagenesis of the lipase cDNA ) Lipase in milk - Physiologic function; Model for studies of endothelial LPL Hormone sensitive lipase (HSL) of adipose - Role of perilipins (expression in adipocytes and steroidogenic cells, PKA-induced phosphorylation); Effect of epinephrine, ACTH, camp; Effect of insulin, prostaglandins and other effectors (adrenoceptors, insulin receptors, and adenosine receptors, differences in subcutaneous and omental adipose sites); Relationship to cholesterol ester hydrolase activity; Studies with HSL cDNA probes; Mechanism of lipolysis and specificity; Pre- and post-translational mechanisms of regulation Hepatic lipase - Gender differences; Role in lipoprotein metabolism; Phospholipase activity vs triglyceride lipase activity (structural basis of enzyme catalysis, use of chimeric lipases to assess substrate specificity); Effects of diets, drugs and hormones; Regulation of activity; Inhibition by phosphonate analogs of triglyceride Lipoprotein lipase (LPL) - Structure and properties; Location, cofactors and action (roles of apolipoproteins, specificity, mechanism of lipolysis, differences in muscle, mammary gland and white and brown adipose, transfer and fate of products); Inactivation and inhibition; Release from endothelium by heparin; Regulation (in vivo and in vitro studies involving effects of diet composition and feeding frequency on enzyme activity, age and gender, diet-induced obesity); Activity of gene in genetic obesity (model: fa/fa rat, leptin); Effects of glucagon, thyroid hormone, catecholamines, insulin and steroid hormones; Action of cAMP, adenosine and inosine; Cold-induced suppression; LPL in pregnancy; Regulation of expression of LPL mRNA in concert with VLDL receptor mRNA (gene transfer experiments, effect of endothelin-1, oxysterol inhibition of LPL mRNA synthesis, post-transcriptional regulation) Phospholipases - Phospholipase A1 (occurrence, specificity, and role in sequential deacylations, stimulation by activation of 2-adrenergic receptors, platelet phospholipase A1/lysophospholipase activity); Phospholipase A2 (occurrence, specificity and importance, differences in type 1 and type 2, regulation of expression, activation by melittin); Phospholipase A2 activating protein (PLAP) (role in activation of PKC-MAP kinase cascade, regulation of expression); PAF acetylhydrolase (a specific phospholipase A2 specificity, comparison with other phospholipases A2); Generation of eicosanoids (effect of release of phospholipase A2 into circulation in shock, sepsis and injury, importance in therapy of inflammation and in profound disease states); Modulation of activity (dexamethasone, late pregnancy and parturition, relationship to TNF and apoptosis, overexpression of A20 zinc finger protein, effects on activation by protein kinase A inhibitors, increased activity (type 2) in Crohn's Disease and in colon neoplasms, chemical inhibitors); Phospholipase C - Phosphoinositide-specific enzyme (roles in signal transduction, relationship to G-protein family and Ca++ mobilization, effects of endothelin-1, interleukins and TNF ,importance in platelet and neutrophile functions, PDGF-derived induction of activity, sphingosine-phosphate induction of enzyme activity); Phospholipase C - Phosphatidylcholine-specific enzyme; Phospholipase D (activation by ceramide, activation by oxidized LDL and relationship to smooth muscle cell proliferation, induction of activity by endothelin-1, PAF and other agonists, synergistic activity with phospholipase C, specific effects in different cell types, role in release of phosphatidic acid as a second messenger, role of phosphatidylcholine-specific phospholipase D in transduction, membrane trafficking, and cell proliferation) Sphingomyelinase - Role in formation of ceramide as a second messenger; Chlorpromazine and other inhibitors of enzyme activity Ceramidase Arylsulfatase A - Role in hydrolysis of sulfatides to galactosyl-ceramide; Sphingolipid activator proteins

Sphingolipid ceramide deacylase Disorders - Partial or complete deficiency of lipoprotein lipase (biochemical abnormalities, symptoms and sequelae); Sphingolipid storage diseases (Gm1 gangliosidosis, Tay-Sachs disease, Fabry's disease, Sandhoff's disease, Gaucher's disease, Niemann-Pick disease, Farber's lipogranulomatosis, Krabbe's disease, Sulfatide lipidosis); Phospholipid disorders (Lowe syndrome, Miller-Dieker syndrome, Choline deficiency)

Metabolic Fates of Cholesterol

Overview Cholesterol as a structural component - Locations in cell; Differences in content among tissues; Intracellular transport; Characteristics of cholesterol-loading in erythrocyte membranes Importance of esterification reactions - ACAT reaction; LCAT reaction; Cholesterol esterase Synthesis of steroid hormones - Cellular cholesterol synthesis vs. plasma transport of cholesterol into steroid-secreting endocrine cells; Major enzymes in the conversion to hormones (side chain cleavage enzymes, cytochrome P450 enzymes, 17-hydroxylase and other hydroxylases, 18-oxidase, aromatase, regulation of conversion) Formation of vitamin D3 - Conversion of cholesterol to 7-dehydrocholesterol; Hydroxylation reactions in liver and in kidney; General features of regulation of synthesis Formation of bile acids and secretion of bile - Cholesterol-7-hydroxylase (chemistry of reaction, control of activity, importance of enzyme to cholesterol metabolism); Hydroxylation of C-12; Cleavage of side chain and conversion of C-24 to carboxyl; Conjugation and sulfation reactions; Conversion to secondary bile acids; Bile composition (bile acid micelle and its function, cholesterol: phospholipid: bile acid ratios, importance in elimination of cholesterol) Oxidized cholesterol - Dietary oxysterols, sources and metabolism; In vivo formation of cholesterol oxides (products, physiologic relevance and pathologic consequences) Implications of very low plasma cholesterol levels Dietary phytosterols - Major types; Absorption of phytosterols and effects on cholesterol absorption; Metabolic fate Disorders related to cholesterol metabolism - Familial LCAT deficiency; Malnourishment and amenorrhea; Rickets; Cholelithiasis; Cholesterol ester storage disease; Hereditary -sitosterolemia; Wolman's disease; Cerebrotendinous xanthomatosis

Eicosanoid Metabolism
Overview Nomenclature - Prostaglandins A, D, E, F, G, H, I, J; Series 1, 2, or 3 prostaglandins; and designations; Leukotrienes A, B, C, D, E, F; Hepoxilins (hepoxins) and lipoxins; Isoprostanes Generation of precursor fatty acids from phospholipids - Phospholipase A2 activation and action (locations of enzymes, stimuli for release of fatty acids, anti-inflammatory action of corticosteroids); Phospholipase C and DAG lipase activation and actions Prostaglandin endoperoxide synthetase (PES) - Location and characterization of enzyme complex (structural resemblance to epidermal growth factor, cyclooxygenase); Gene expression in different tissues; Activation of enzyme; Chemistry of catalysis; Products from different precursor fatty acids (dihomo-linolenic acid, arachidonic acid, eicosapentaenoic acid); Products of peroxidase activity; Inhibition by non-steroidal antiinflammatory drugs (NSAIDs): Functions of products (comparison of products of the 1, 2, 3 series, half-life of products and target tissues, effects on various cells, prostaglandin receptors, ligand for the peroxisomal proliferator activated receptor (PPAR)); Effects of docosahexaenoic acid on prostaglandin synthesis Isomerase and reductase actions Thromboxane A2 synthetase - Regulation of enzyme activity; Cloning and characterization of the gene; Regulation of synthesis in megakaryocytes and platelets (dependence on precursor release and on PES activity); Protein kinase Cdependent induction of synthesis in macrophages; Induction in monocytes by lipopolysaccharide or phorbol ester, Post-transcriptional regulation; Thromboxane A2 receptors; Functions of thromboxane A2 (normal physiology, pathophysiology of excess formation); Inhibitors of synthase Lipoxygenases - Locations and characterizations of enzymes; Mechanism of action (iron-sulfur complex activity, formation of radicals); 5-Lipoxygenase (5-LO) (action of glutathione on enzyme activity, products from different fatty acid precursors, 5-LO activating protein (FLAP) and its mechanism of action and inhibitors); 12-Lipoxygenase (12LO) (formation and metabolism of hepoxilins, actions of hepoxilins); 15-Lipoxygenase (15-LO) 5-, 12-, or 15-Hydroxyperoxyeicosatetraenoic acids (HPETEs) - Conversion to hydroxyeicosatetraenoic acids (HETEs); Synthesis of DiHETEs; Synthesis of leukotriene A4 (LTA4); Products following glutathione transferase activity (formation of LTC4, formation of LTD4 slow reacting substance of anaphylaxis); LTB4 formation via hydrolase; Regulation of synthesis of leukotrienes; Functions of leukotrienes Non-enzymatic, free-radical formation of isoprostanes Effects of excess dietary omega-3 fatty acids on prostaglandin and leukotriene pathways - Epidemiologic studies; Human clinical studies; Animal studies

Disorders - Contribution to normal and abnormal smooth muscle contraction (parturition and abortion, asthma, blood pressure); Influence on platelet aggregation and thrombosis; Anaphylaxis; Hypersensitivity reactions and inflammatory diseases; Hereditary PGH synthetase deficiency in platelets

Transfer and Transport of Lipids

Overview The calycin superfamily Fatty acid binding proteins (FABP) - Structure and properties (differences among tissues, relationship to other calycins); Functions (developmental role in nervous system, role in placental transfer of fatty acids, cellular uptake and metabolism of fatty acids, differences in affinity for specific fatty acids); Regulation of synthesis (dietary effects, cloning studies on expression of FABP mRNA Lipocalins - Occurrence and functions; Relationship to apolipoprotein D; Possible role in mediation of cell growth Sterol transport proteins (sterol carrier proteins) - Intracellular transport - sterol carrier protein 2 (SCP2) (structure and properties, cholesterol and phospholipid transfer activity, tissue distribution, roles in lipid absorption by intestinal cells, in transfer in steroid synthesizing cells and in use of transfected mouse cell models for studies, in vitro affinity studies with recombinant SCP2, mediation of expression in ovarian cells by PGF2 ; Role of apolipoprotein E in intracellular sterol transport (structure comparison to SCP2, expression in steroidogenic tissues); Sterol and phospholipid transfer activity of 17-hydroxysteroid dehydrogenase; FABP influence on sterol transfer; Extracellular sterol transport facilitated by apo AI Lipid transfer protein/Lipopolysaccharide binding protein family - Cholesterol ester transfer protein (CETP) (molecular structure, specificity, function and mechanism, regulation of CETP gene expression on enzyme activity, genetic deficiency of CETP, CETP transgenic mice, CETP relationship to atherogenesis); Phospholipid transfer protein [PLTP] (Structure, specificity, function and mechanism, PI/PC transfer protein studies in vitro, PLTP mediation of HDL metabolism); Regulation of activity (promotor region of human PLTP gene, effect of lipid lowering drugs); Comparative relationships to lipopolysaccharide binding protein Acyl cholesterol acyl transferase (ACAT) - Role in intracellular cholesterol homeostasis (substrate specificities, tissue distribution); Effect of ACAT inhibitors in macrophages; Relationship to apolipoprotein B and VLDL secretion; Regulation of activity (cloning and expression of mouse ACAT, effect of steroid hormones, dietary effects, effect of pharmaceutical hypocholesterolemic agents, effect of heparin, model: transfected Chinese hamster ovary cell lines) Lecithin cholesterol acyl transferase (LCAT) - Structure and function (specificity in preferential acylation of pregnenolone, acylation of cholesterol and other steroids, fatty acid preference, Apo A-I cofactor dependency); Regulation of activity (gender differences in animals, tissue distribution, effect of hypo-cholesterolemic drugs); Inhibition (effect of lipid peroxidation on modification of apo A-I, specific chemical inhibitors and significance); Role in "reverse cholesterol transport" and HDL remodeling Disorders - Niemann-Pick type C: Familial combined hyperlipidemia; Familial LCAT deficiency

Adipocyte Development; White and Brown Adipose Tissue

Overview Preadipocyte cell lines Sequence in differentiation Role of C/EBP and control of its expression by glucocorticoids and insulin; Hormoneresponsive transcription factors (interactions with adipose response elements (AREs) in specific genes, modulation by inducers: IGF-1, glucocorticoid, cAMP, long chain fatty acids; PPAR nuclear receptor ( fatty acids as ligands, natural prostaglandin ligand (15d-PGJ2), antidiabetic thiazolidione drugs); Coordinate gene expression and transcriptional activity (422/aP2 Fatty acid binding protein, fatty acid activated receptor (FAAR), adipocyte determination and differentiation factor-1 (ADD-1), preadipocyte repressor element); Acylation stimulating protein (ASP) (adipsin-ASP pathway, effect on membrane transport of glucose, effect on DAG acyltransferase, role in pathogenesis of obesity); Effects of hormones and neurotransmitters (serotonin, dopamine and neuropeptide y, cholecystokinin, dehydroepiandrosterone, leptin, cytokines, "obesity drugs", dietary studies with "fake fats") White vs. brown adipose (WAT, BAT) Locations ; Thermogenesis (obligatory, facultative, and occurrence in lactation, fasting or hibernation) Regulation of body weight - Satiety factor hypothesis; Intake of specific nutrients hypothesis; Cafeteria diet hypothesis; High fat diet hypothesis; Diet-induced obesity (animal studies, human studies); Genetic obesity (human twin studies, animal models - Ob/ob gene - a defect in leptin, tubby gene, fa/fa model, JCR:LA-corpulent rats); Studies using brain lesions (paraventricular nucleus (PVN) lesions, ventromedial hypothalamus (VMH) lesions, lateral hypothalamic (LH) lesions); Hormonal influences Disorders - Consequences of morbid obesity; Prader-Willi syndrome; Relevance of obesity to development of noninsulin-dependent diabetes mellitus

Lipoproteins
Overview Triglyceride-rich lipoproteins - Formation of nascent chylomicrons and intestinal very low density lipoproteins (VLDL); Lipid compositions and requirements for secretion; Synthesis of apolipoproteins; Assembly and secretion of particles; Catabolism of chylomicrons and intestinal VLDL (role of lipoprotein lipase (LPL), role of apolipoprotein E (apo E)); Hepatic chylomicron remnant receptors (structure and characteristics, regulation, physiologic relevance); Half-life of particles; Fates of various components; Hepatic VLDL (composition and influence of diet, synthesis of components, assembly and secretion, role of LPL in metabolism of VLDL); Roles of lipid transfer proteins in metabolism (phospholipid transfer, cholesterol ester transfer, importance of LCAT); general features of the conversion to intermediate and low density lipoproteins (IDL, LDL); Endothelial VLDL receptor (binding of apoE-rich lipoproteins and receptor-associated protein (RAP), binding of LPL and urokinase/plasminogen activator inhibitor, importance in regulation of fibrinolysis, release of IDL/LDL from binding site); Hepatic B/E receptor uptake; -VLDL receptor uptake of abnormal VLDL particles; Methods of studying VLDL kinetics of metabolism Low density lipoproteins - Composition and characteristics of LDL of varying sizes; Normal kinetics of metabolism (apolipoprotein B (apo B) tracer use in modeling studies, lipid tracers); Functions and importance of LDL; LDL receptor-mediated uptake; Locations and characterization of receptor; Mechanism of uptake (requirements for binding ligands, movement into clathrin-coated "pits", model of receptor recycling); Regulation of receptor expression (cloning studies, transgenic animal studies, mutant receptor studies in vitro) Sterol-regulatory element; Fates of intracellular LDL degradation products; LDL oxidation in arterial wall (possible reactions, predisposing conditions, consequences) High density lipoprotein (HDL) - Synthesis of components and secretion of nascent HDL; Compositions and characteristics of HDL subclasses; Intravascular metabolism of HDL and conversion of subclasses; Lipid transfer reactions; Formation of spherical HDL; Theory of reverse cholesterol transport by HDL (basis of theory, animal, human and cell culture studies); Catabolism of HDL; HDL receptors (hepatic, macrophage, other); Fates of components Comparisons of lipoprotein receptors - Types and distributions (chylomicron remnant receptors, -VLDL receptors, B/E receptors, LDL receptors, acetyl-LDL and other modified-LDL receptors, HDL receptors); Ligands and required cofactors; Structural relationships; General aspects of regulation (hormones, sterols and oxysterols, PDGF and other growth factors, HMG CoA reductase inhibitors and bile acid resins) Apolipoproteins - Types, lipoprotein associations and sites of synthesis; Apolipoprotein gene family; Relevance to dyslipidemias; Apo A-I, A-II and A-IV (cDNA clones and location of genes, regulation of expression, genetic variants, functions); Apo B48 and B100 (Chromosomal location and cDNAs, genetic variants, regulation of expression n intestine, constitutive expression in hepatocytes, regulation of assembly into lipoproteins, functions and catabolism); Apo C-I, C-II, C-III (characteristics, clones and gene structures, genetic mutations, regulation of expression, functions and catabolism); Apo E (characteristics, clones and gene structures, genetic mutations, regulation of expression, functions and catabolism, link to Alzheimer's disease?); Minor apolipoproteins Lipoprotein(a) - Physical, chemical and biological properties; Apo (a) (Kringle domain, serine protease domain, homology to plasminogen); Apo B; Lipids; Synthesis; Catabolism; Genetics; Atherogenesis HDLc formation and metabolism Integration of overall lipoprotein metabolism - Roles of LCAT, ACAT, LPL, CETP, PLTP; Influence of diet; Effects of hormones (insulin, growth hormone, thyroid hormone, steroid hormones, other) Use of human hepatoma cells to study lipoprotein metabolism - Types of particles secreted; Efficiency of lipoprotein catabolism; Pitfalls of use as a model of liver Disorders Abetalipoproteinemia; Tangier's disease; Type I hyperlipidemia; Familial LPL deficiency; Apo C-II deficiency; Type IIa hyperlipidemia (hypercholesterolemia) (LDL receptor defects, polygenic hypercholesterolemia); Type IIb hyperlipidemia (familial multiple hyperlipidemia); Type III hyperlipidemia (familial dys--lipoproteinemia); Type IV hyperlipidemia (familial hypertriglyceridemia (mild), sporadic hypertriglyceridemia, association with obesity, alcohol intake and NIDDM); Type V hyperlipidemia (mixed hyperlipidemia) (familial hypertriglyceridemia (severe), familial LPL deficiency, apo C-II deficiency); Secondary hyperlipidemias (uncontrolled diabetes mellitus, hypothroidism, systemic lupus erythematosus, nephrotic syndrome, Cushing's syndrome, alcoholism, uremia, liver disease with biliary stasis, hypopituitarism, stress, various drugs and hormone preparations); Implications for diet therapy; Other therapeutic approaches (drugs, gene therapy, liver transplants)

Lipid Oxidation and Antioxidants

Overview Sequence of events in lipid peroxidation - Generation of oxygen radicals; Nonenzymatic peroxidation (formation of peroxyl radical, formation of hydroperoxide, products); Enzymatic peroxidation (lipoxygenase and cyclooxygenase, hydrogen peroxide and superoxide anion production and roles in peroxidation of lipids); Role of iron and other metals in peroxidation (Fenton reaction, biological sources of ions); Other methods of generation of oxygen radicals (radicals derived from drugs and chemicals, pulmonary hyperoxia)

Measurement of peroxidation - Conjugated dienes, thiobarbituric acid assay, breath alkanes, chromatography methods: Advantages and disadvantages of these assays; Lipid oxidation products in foods Cellular injury and its assessment - Meaning of "oxidative stress"; Biochemical lesions, DNA-adduct formation, Aldehyde modification of amino groups, peroxidation of fatty acids and cholesterol); Types of damage (endothelial injury, reperfusion injury of myocardium and other sites, inflammatory diseases, effects on aging?, nephrotoxicity); Cell mediated oxidation of LDL (proposed mechanism, significance) Antioxidant nutrients and other compounds - Direct scavengers and their reactions (Vitamin E, Vitamin C, Carotene); Antioxidant enzymes and their essential minerals (superoxide dismutase, glutathione peroxidase, catalase); Other antioxidant compounds (Ubiquinone-10, Dihydrolipoic acid, tocotrienols and similar quinines, flavonoids, Probucol and other drugs); Differences in oxidative susceptibility of plasmalogens and other phospholipids; Influence of alcohol consumption; Studies on antioxidants and various dietary oils (animal and human studies) Relationship of Lipid Metabolism to Atherosclerosis and Coronary Heart Disease Overview Stages of atherogenesis - Fatty streak formation; Atheroma formation; Calcification and plaque formation; Sequelae: Compromised circulation and infarction Risk factors for development of atherosclerosis and CHD Genetics; Hypertension and its relationship to other factors; Gender and sex hormone relationships to other factors; Smoking; Hypercholesterolemia (National Cholesterol Education Program, current recommendations for clinical cut-points, relationship to other factors); Hypertriglyceridemia; Diabetes mellitus; Other factors; Interactions of risk factors Theories of atherogenesis and supporting evidence - Response to endothelial injury hypothesis (mechanical damage, damage by microbial or viral infections, antigen-antibody complex, lipid peroxides, homocysteine, xenobiotic and other chemicals, elevated plasma cholesterol); Monoclonal cell hypothesis; Current "unified" theory Overall model of atherogenesis - Sequence of events after initiation of intimal injury (role of elevated lipids, roles of monocyte/macrophages); Smooth muscle cell proliferation; PDGF and other growth factors; Role of modified LDL and the macrophage scavenger cell receptor; Influence of leukocytes and leukocyte factors; Secretion and actions of interleukins, PAF, TNF and adhesion molecules; Contribution of hyperlipidemia; Contribution of oxidized LDL; Factors affecting LDL interaction with arterial matrix components Further development of atheroma and lipid accumulation Thrombogenesis - Platelet aggregation and adhesion; Increase in thromboxane A2 and serotonin levels; Promotion of vasoconstriction and cell proliferation Impact of hypertension and diabetes mellitus Studies on regression of stenosis Studies on relationship of risk factors, diets and coronary heart disease - Animal studies (high/low fat diets, manipulation of fiber, type fat or other dietary components, transgenic animal studies); Human studies (Epidemiological studies retrospective, intervention, prospective, "Clinical" trials, at-risk individuals, public health approach to prevention and rationale)

Dietary Fats: Uses and Abuses

Overview Caloric intake and need for fat in the diet Alterations in dietary SF:MUF:PUF ratio - General effects; Differences in needs of population groups; Impact of ratio of omega-9, -6, -3 dietary fatty acids - Human epidemiological and clinical studies; Animal studies; Effects on lipoprotein composition, structure, and biological actions (hypotriglyceridemic effect of fish oils, alteration of ratio of TC/HDL-C; concerns about high intakes of fish oils) Importance of chain length of dietary SFA - Impact on absorption; Impact on in vivo cholesterol synthesis Trans fatty acids - Metabolism and fate; Impact on lipoproteins and risk of cardiovascular disease; Current dietary recommendations Conjugated diene fatty acids in the diet - Metabolism and fate; Implications for health (claims for anti-carcinogenic effect and antiatherosclerosis effects of conjugated linoleic acid) Therapeutic uses of gamma-linolenic acid Relationships between dietary fat and the immune response - Review of biochemical relationships; Impact of amount, as well as type, of dietary fat; PUF influence on cytokine synthesis Relationships between dietary fat and brain function - The developing nervous system and special needs; FA therapy in schizophrenia, clinical depression; Theory of very low cholesterol levels and brain function Relationships between dietary fats and certain chronic diseases - Insulin resistance; Obesity; Tumors and cancers; Arthritis; Atherosclerosis; Immune diseases; Genetic diseases Survey of health claims and known effects of specific dietary oils - Cottonseed, palm and soybean oils; Sunflower, safflower, corn and peanut oils; Canola, olive and other high oleic oils; Sesame seed, flaxseed and evening primrose oils; Oils from nuts; Menhaden and other fish oils; Mead acid (n-9 eicosatrienoic acid); Rice, barley, oat and amaranth oils

Impact of fat substitutes, low fat and low cholesterol foods on dietary habits and human health