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Bertholf, Ph.D. Associate Professor of Pathology Director of Clinical Chemistry & ToxicologyffClassification of lipidsFatty acids (palmitic, linoleic, etc.) Glycerol esters (triglycerides) Sterols (cholesterol, hormones, vitamin D) Terpenes (vitamins A, E, K) Sphingosine derivatives (sphingomyelin)t  -    Fatty acidsEven-numbered fatty acids predominate The most common saturated fatty acids are palmitic (16:0) and stearic (18:0), but unsaturated fatty acids are more common in natureZ,P  > Unsaturated fatty acids2Double bonds in fatty acids are nearly always cis $3.. Essential fatty acidsMammals can synthesize saturated and mono-unsaturated fatty acids. Linoleic (18:2) and linolenic (18:3) fatty acids cannot be synthesized, and therefore must be obtained from the diet (plants). Both are required for the biosynthesis of prostaglandins&99>C   "Clinical importance of fatty acidsJFecal fatty acids are sometimes measured to detect malabsorptive and pancreatic disorders the test is mostly considered obsolete Serum free fatty acids help distinguish between hyperinsulinemic hypoglycemia (FFA normal) and disorders of fatty acid oxidation (FFA elevated and negative ketones)&Z&>3p[Glycerol esters (acylglycerols) ~Triglycerides are the most abundant family of lipids in plant and animal cells, and are major components of the the human dietq*Measuring triglycerides (reference method) =Triglycerides are extracted into chloroform prior to analysis0*Measuring triglycerides (enzymatic method)   Sterols (cholesterol)Sterols are steroid backbones that have a hydroxyl group at position 3 and a branched aliphatic chain of 8 or more carbons at position 17 Cholesterol biosynthesis]About 2% (approximately 1 g) of total body cholesterol is replenished each day Dietary sources account for less than half Cholesterol is synthesized from Acetyl CoA 90% of in vivo synthesis occurs in the intestine and liver (although all cells have the capability) Absorption of dietary cholesterol appears to have a maximum of approximately 1 g/dayHOUO]VU Cholesterol biosynthesis Measuring cholesterol by L-BThe Liebermann-Burchard method is used by the CDC to establish reference materials Cholesterol esters are hydrolyzed and extracted into hexane prior to the L-B reactionZ,   Enzymatic cholesterol methodsEnzymatic methods are most commonly adapted to automated chemistry analyzers The reaction is not entirely specific for cholesterol, but interferences in serum are minimalZ LipoproteinszIn order to be transported in blood, lipids must combine with water-soluble compounds, such as phospholipids and proteins.{Z{_ Lipoprotein classes2%Appearance of hyperlipidemiajStanding Plasma Test for chylomicrons Plasma is placed in refrigerator (4C) overnight Chylomicrons accumulate as floating  cream layer Chylomicrons in fasting plasma are abnormal @&> 1 %  1$Lipoprotein electrophoresisLEP is no longer a common laboratory test Standing plasma test for chylomicrons Total cholesterol, TG, HDL, and LDL can be measured directly**ZcZ*cC >8*Fredrickson classification"Measuring HDL cholesterolUltracentrifugation is the most accurate method HDL has density 1.063  1.21 g/mL Routine methods precipitate apolipoprotein B with a polyanion/divalent cation Includes VLDL, IDL, Lp(a), LDL, and chylomicrons j0"N10"N1lO  #Indirect LDL cholesterolXFriedewald formula assumes that all cholesterol is VLDL, LDL, and HDL lipoproteins Chylomicrons are usually low in normal, fasting subjects, and IDL and Lp(a) are usually insignificant contributors to total cholesterol Since VLDL is 55% TG and 12% Chol: [LDL Chol] = [Tot Chol]  [HDL Chol]  [TG]/5 XS#.S#/~ I :]   $Direct LDL cholesterolOlder direct methods for LDL involved precipitation with heparin or polyvinyl sulfate Newer methods involve precipitation of VLDL, IDL, and HDL with polyvalent antibodies to Apo A and Apo E LDL is almost exclusively Apo B-100&$$>%Direct vs. Indirect LDL2The Friedewald equation assumes that chylomicrons, IDL, and Lp(a) are not significant Non-fasting specimens can have chylomicrons TG > 400 mg/dL indicates the presence of chylomicrons (or remnants) Type III hyperlipidemia is characterized by high b-VLDL, which has a 3:1 TG:C ratioHVpTVp1"   7   <Apolipoproteins~The protein composition differs from one lipoprotein class to another, and the protein constituents are called Apolipoproteins$ooFunctions of apolipoproteins Activate enzymes involved in lipid metabolism (LCAT, LPL) Maintain structural integrity of lipid/protein complex Delivery of lipids to cells via recognition of cell surface receptors Apolipoprotein content of LPs*#Cholesterol metabolism (exogeneous)$$( !$Cholesterol metabolism (endogeneous)%%( &DyslipoproteinemiasCauses can be primary or secondary Secondary causes include starvation, liver disease, renal failure, diabetes, hypothyroidism, lipodystrophies, drugs Primary causes of hyperlipidemia: Increased production Defective processing Defective cellular uptake Inadequate removalT#t"W#t"W,Y'Dyslipoproteinemias7Hyperchylomicronemia LPL deficiency Apo C-II deficiency&##$Hyperchylomicronemia(- DyslipoproteinemiasHyperchylomicronemia LPL deficiency Apo C-II deficiency Hyperbetalipoproteinemia Overproduction of VLDL Enhanced conversion of VLDL to LDL LDL enriched with cholesteryl esters Defective LDL structure Decreased LDL receptors\##HM 7,Hyperbetalipoproteinemia(+DyslipoproteinemiasWCombined hyperlipoproteinemia Normal LDL receptors Overproduction of VLDL and Apo B-100&::, 0."Combined hyperlipoproteinemia( /!DyslipoproteinemiasCombined hyperlipoproteinemia Normal LDL receptors Overproduction of VLDL and Apo B-100 Dysbetalipoproteinemia Both cholesterol and triglyceride elevated Mutant form of Apo E T:A:Ab 0 0#Dysbetalipoproteinemia(3&DyslipoproteinemiasWFamilial hypercholesterolemia Defect in LDL receptor Absent Defective Incidence = 1:500N :4'Familial hypercholesterolemia( 5(DyslipoproteinemiasFamilial hypercholesterolemia Defect in LDL receptor Absent Defective Incidence = 1:500 Familial defective Apolipoprotein B-100r(( , M6)Familial hypercholesterolemia( 9+High cholesterol, high LDLDiet/Lifestyle 2 to hypothyroidism or nephrotic syndrome (disruption of Apo-B metabolism) Polygenic: (means we don t know) Familial hypercholesterolemia Familial defective Apo-B Rare disordersb'  !:,High TG, normal cholesterolDiet/Lifestyle 2 to diabetes, thiazide diuretics, Cs, beta-blockers, CRF/Nephrotic syndrome Familial hypertriglyceridemia (etiology unknown) ApoC-III excess (interferes with LPL) LPL deficiency ApoC-II deficiencyZb# 1;-High cholesterol, TGObesity 2 to steroids, Cs, hypothyroidism, CRF Familial combined hyperlipidemia (multifactorial) Peroxisome proliferator-activator receptor Dysbetalipoproteinemia (Type III) Hepatic lipase deficiency 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&՜.+,0X    On-screen ShowHn-sZ/ 6Times New RomanTimesArialSymbol Arial Black CHALKBOACS ChemDraw DrawingLipids and LipoproteinsClassification of lipids Fatty acidsUnsaturated fatty acidsEssential fatty acids#Clinical importance of fatty acids Glycerol esters (acylglycerols)+Measuring triglycerides (reference method)+Measuring triglycerides (enzymatic method)Sterols (cholesterol)Cholesterol biosynthesisCholesterol biosynthesisMeasuring cholesterol by L-BEnzymatic cholesterol methods LipoproteinsLipoprotein classesAppearance of hyperlipidemiaLipoprotein electrophoresisFredrickson classificationMeasuring HDL cholesterolIndirect LDL cholesterolDirect LDL cholesterolDirect vs. Indirect LDLApolipoproteinsFunctions of apolipoproteinsApolipoprotein content of LPs$Cholesterol metabolism (exogeneous)%Cholesterol metabolism (endogeneous)DyslipoproteinemiasDyslipoproteinemiasHyperchylomicronemiaDyslipoproteinemiasHyperbetalipoproteinemiaDyslipoproteinemiasCombined hyperlipoproteinemiaDyslipoproteinemiasDysbetalipoproteinemiaDyslipoproteinemiasFamilial hypercholesterolemiaDyslipoproteinemiasFamilial hypercholesterolemiaHigh cholesterol, high LDLHigh TG, normal cholesterolHigh cholesterol, TG Low cholesterol, low/normal HDLLow HDL High HDL  Fonts UsedDesign TemplateEmbedded OLE Servers Slide Titles/&_;%Roger BertholfRoger Bertholf  !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~      !"#$%&'()*+,-./0123456789:;<=>?@ABCDEFGHIJKLMNOPQRSTUVWXYZ[\]^_`abcdefghijklmnopqrstuvwxyz{|}~Root EntrydO)PicturesCurrent UserSummaryInformation(`RPowerPoint Document(;DocumentSummaryInformation8