Medical Technology Board Examination Review Notes Must Know (Rodriguez) 2

Specimen Collection and Handling
Sleeping patients	Must be awakened before blood collection
Unconscious patients	Ask nurse or relative Identification bracelet
Venipuncture	Median Cubital (1st) à Cephalic (2nd) à Basilic (3rd)
Tourniquet	Velcro or Seraket type 3-4 inches above the site Not exceed 1 minute
Needle	Bevel up 15-30O angle Length: 1 or 1.5 inch (Butterfly needle: ½ to ¾ inch)
After blood collection	Cotton à site Apply pressure for 3-5 minutes
BP cuff as tourniquet	Inflate to 60 mmHg
Benzalkonium chloride (Zephiran)	Disinfectant for ethanol testing Dilution – 1:750
IV line on both arms	Discontinue IV for 2 minutes Collect sample below the IV site Initial sample (5mL) à discard
IV fluid contamination	Increased: Glucose (10% contam. w/ 5% dextrose à increased bld glucose by 500 mg/dL) Chloride Potassium Sodium Decreased: Urea Creatinine
Renin blood level	Collected after a 3-day diet, from a peripheral vein
Basal state collection	Early morning blood collection 12 hours after the last ingestion of food
Lancet	1.75mm: preferred length to avoid penetrating the bone
Incision (Skin puncture)	<2.0mm (infants and children) 2-3mm (adults)
1.5-2.4mm	Distance from the skin surface to bone or cartilage (middle finger)
Arterialized capillary blood	Earlobe: Preferred site Lateral plantar heel surface: most commonly used site
Flea	Minute metal filling which may be inserted into the capillary tube before collecting blood to help mix the specimen while the blood is entering the tube
Indwelling umbilical artery	Best site for blood gas analysis (newborns)
1000-3000 RCF for 10 mins	Centrifugation requirement
Hemolysis	Increased: “KLA6MP ITC2” -K+ -LDH (150x) -ACP -ALP -Aldolase -ALT -AST -Albumin -Mg2+ -Phosphorus -Iron -Total protein Affects bilirubin levels Inhibits lipase
Refrigeration/Chilling (Low temp)	Required for: “ABCGLRP2” Ammonia Blood gases Catecholamines Gastrin Lactic acid Renin PTH Pyruvate Decreased: LD 4 and 5 Increased: ALP
Photosensitive analytes	Bilirubin Beta-carotene Folate Porphyrins Vitamins  A and B6
Oxalate	Insoluble salt 1-2 mg/mL blood
Citrate	Non-ionized form 3.2-3.8 g/dL (1:9 ratio)
EDTA	Chelation 1-2 mg/mL blood Versene: disodium salt Sequestrene: Dipotassium salt
Fluoride	Weakly dissociated calcium component 2 mg/mL blood: anti-glycolytic 10 mg/mL blood: anticoagulant
Heparin	A.k.a. Mucoitin polysulfuric acid Universal anticoagulant Antithrombin 0.2 mg/mL blood
Lithium heparin	For glucose, BUN, ionized calcium, electrolyte studies (K+: best) and creatinine
Orange top tube	Additive: Thrombin
Royal blue top tube	Additives: None; Na2EDTA Sodium heparin
Brown top tube	Lead testing
Tan top tube	Lead testing
Black top tube	Additive: Buffered sodium citrate For ESR
Respinning gel tubes	Increases potassium
Thixotropic gel	Gel separator (SG: 1.04) Serum: (SG: 1.03) RBC: (SG: 1.05)
Laboratory Mathematics
% w/v	Grams of solute = % solution desired x total volume desired                                                                    100
% v/v	mL of solute = % solution desired x total volume desired                                                              100
% w/w	Grams of solute = % solution desired x grams of the total solution                                                                        100
Molarity	M =          _grams of solute_______          GMW x volume of solution
Moles	Mol = weight (grams)                    GMW
To prepare a molar solution	Grams of solute = Molarity x GMW of the solute x Volume (L) desired
To convert % w/v to Molarity	M = % w/v Ÿ 10               GMW
Normality	N = _Grams of solute_         EW x volume (L)
Equivalent weight (EW)	EW = __MW___             valence
To prepare a normal solution of solids	Grams of solute = Normality x EW x Volume (L)
To convert % w/v to Normality	N = w/v Ÿ 10             EW
Normality	N = Molarity x Valence
Molarity	M = Normality           valence
Molality	m =    Grams of solute__         MW x kg of solvent
Milliequivalents	mEq/L = mg/dL Ÿ 10 Ÿ valence                                  MW
Millimoles	mmol/L = mg/dL Ÿ 10                             MW
Ratio	Ratio = _Volume of solute_                Volume of solvent
Dilution	Dilution = __Volume of solute__                       Volume of solution
0.179	Conversion factor for iron (mg/dL à μmol/L)
0.01	Conversion factor for phospholipid (g/dL to g/L)
2.27	Conversion factor for folate
Analytical reagent (AR) grade	For qualitative and quantitative analyses For accuracy Established by American Chemical Society (ACS) Uses: Trace metal analysis and preparation of standard solutions
Ultrapure reagents	Additional purification steps Ex: Spectrograde, nanograde, HPLC grade Uses: Chromatography, atomic absorption, immunoassays
Chemically Pure (CP) or Pure Grade	Indicates that the impurity limitations are not stated Purity is delivered by meas. of melting point or boiling point
Technical/Commercial grade	In manufacturing Never used in clin. lab. testing
United States Pharmacopoeia (USP) and National Formulery (NF)	For human consumption Not applicable for lab. analysis Purpose: For drug manufacturing
Preparation of reagent grade water	Filtration (1st) à Distillation, Ion exchange, Reverse Osmosis
Type I Rgt Water	Min. interference Max. water purity Used immediately For ultramicrochemical analyses, measurements of nanogram or subnanogram concentrations, tissue or cell methods (microscopy) and preparation of standard solutions Uses: FEP, AAS, blood gases and pH, enzyme studies, electrolyte testing, HPLC, trace metal and iron studies
Type II Rgt Water	For clinical laboratory use (hematology, microbiology, immunology, chemistry) For prep. of rgts and QC materials
Type III	For washing glasswares For urinalysis, parasitology and histology
Distilled water	Purified to remove almost all organic materials
Deionized water	Free from mineral salts; removed by ion exchange processes Organic material may still be present
Occupational Safety and Health Act (OSHA)	Req. manuf. to indicate lot no., physical or biological health hazard of the chem.. rgts, and precautions for safe use and storage
College of American Pathologists (CAP)	Recommends that a lab. document culture growth, pH and specific water resistance on reagent grade water
Tests for water purity	Microbiological content pH Resistivity Chemical oxygen demand Ammonia Ions Metals
Detergent-contaminated water	Alkaline pH
Hard water	Contains calcium, iron and other dissolved elements
NCCLS	Now: Clinical and Laboratory Standards Institute (CLSI)
Dilute solution	Relatively little solute
Concentrated solution	Large quantity of solute in solution
Saturated solution	Excess of undissolved solute particles
Super saturated solution	Greater concentration of undissolved solute particles than does a saturated solution of the same substance
Primary standard (IUPAC)	Highly purified Measured directly to produce a substance of exact known concentration
Secondary standard	Low purity Concentration is determined by comparison w/ a primary standard
Laboratory Safety
National Fire Protection Association (NFPA) Classification of Fires
Class A fire	Ordinary combustibles: paper, cloth, rubbish, plastics, wood Extinguisher: Water (A), Dry chemical (ABC), loaded steam
Class B fire	Flammable liquids: grease, gasoline, paints, oil Extinguisher: Dry chemical (ABC), carbon dioxide (BC), halon foam (BC)
Class C fire	Electrical equipment and motor switches Extinguisher: Dry chemical (ABC), Carbon dioxide (BC), halon (BC)
Class D fire	Flammable metals: mercury, magnesium, sodium, lithium Extinguisher: Metal X Fought be fire fighters only
Class E fire	Detonation (Arsenal fire) Allowed to burn out and nearby materials protected
Standard Hazards Identification System (Diamond-shaped color coded symbol)
Blue quadrant	Health hazard
Red quadrant	Flammable hazard
Yellow quadrant	Reactivity/Stability hazard
White quadrant	Other special information
Chemical spills	1st step: assist/evacuate personnel
1:10 dilution of chlorine bleach (10%)	To disinfect and clean bench tops In contact with the area for at least 20 minutes HBV: 10 minutes HIV: 2 minutes
Poisonous vapors	Chloroform Methanol Carbon tetrachloride Bromide Ammonia Formaldehyde Mercury
Flammable and combustible solvents	Acetone Ethanol Toluene Methanol Xylene Benzene Isopropanol Heptane
Flammable liquids	Flash point below 37.8OC
Combustible liquids	Flash point at or above 37.8OC
Strong acids or bases	Neutralized before disposal Water should NEVER be added to concentrated acid
Ether	Deteriorate over time à hazardous Forms explosive peroxides
Benzidine	Known carcinogen
Fumehoods	Ventilation: velocity of 100-120 ft/min
Safety showers	Deliver 30-50 gal/min of H2O at 20-50 psi
Carbohydrates
Glycol aldehyde	The simplest carbohydrate
Sucrose	Most common nunreducing sugar
Pancreas	Exocrine: Enzymes (AMS, LPS) Endocrine: Hormones (Insulin, glucagon, somatostatin)
Hyperglycemic Hormones	“GAG CHET” Glucagon ACTH GH Cortisol Human Placental Lactogen Epinephrine Thyroxine
Hyperglycemia (≥126 mg/dL)	Electrolyte Imbalance: Decreased: Sodium, Bicarbonate Increased: Potassium
Hypoglycemia	50-55 mg/dL = Symptoms ≤50 mg/dL = Diagnostic
Whipple’s triad (Hypoglycemia)	Low blood glucose concentration Typical symptoms Symptoms alleviated by glucose administration
6:1	Ratio of BHA to AA in severe DM (Normal = 1:1)
Type 1 DM	IDDM Juvenile Onset Brittle Ketosis-prone 80-90% reduction of beta-cells à Symptomatic Type 1 DM HLA-DR3 and DR4 (+) Glutamic acid decarboxylase (GAD65) (+) Insulin autoantibodies (+) Microalbuminuria: 50-200 mg/24 hours = Diabetic nephropathy (-) C-peptide
Complications of Type I DM	Microvascular disorders: Nephropathy Neuropathy Retinopathy
Type 2	NIDDM Adult type/Maturity Onset Stable Ketosis-resistant Receptor-deficient Insulin resistance: relative insulin deficiency Strong genetic predisposition Geneticist’s nightmare If untreated à glucose: >500 mg/dL à nonketotic hyperosmolar coma
Gestational DM	Screening: 1hr GCT (50g) – bet. 24 and 28 weeks of gestation Confirmatory: 3-hr GTT (100g) Infants: at risk for respiratory distress syndrome, hypocalcemia, hyperbilirubinemia After giving birth, evaluate 6-12 weeks postpartum Converts to DM w/in 10 years in 30-40% of cases
OGTT (GDM)	FBS = ≥95 mg/dL 1-Hr = ≥ 180 mg/dL 2-Hr = ≥ 155 mg/dL 3-Hr = ≥ 140 mg/dL GDM = 2 plasma values of the above glucose levels are exceeded
Impaired fasting glucose (Pre-diabetes)	FBS = 100-125 mg/dL
Impaired glucose tolerance	FBS = <126 mg/dL 2-Hr OGTT = 140-199 mg/dL
FBS	WB = 15% lower than in serum or plasma VB = 7 mg/dL lower than capillary and arterial blood
CSF glucose	60-70% of the plasma glucose
Peritoneal fluid glucose	Same with plasma glucose
Plasma glucose increases w/ age	Fasting: 2 mg/dL/decade Postprandial: 4 mg/dL/decade Glucose challenge: 8-13 mg/dL/decade
w/in 1 hour (Preferably w/in 30 mins)	Separate serum/plasma from the cells
5-7%/hr	Glycolysis at room temperature
1-2 mg%/hr	Glycolysis at refrigerated temperature
Copper reduction methods	Cupric à Cuprous à Cuprous oxide
Folin Wu	Cuprous ions + phosphomolybdate à phosphomolybdenum blue
Nelson-Somogyi	Cuprous ions + arsenomolybdate à arsenomolybdenum blue
Neocuproine method	Cuprous ions + neocuproine à Cuprous-neocuproine complex (yellow)
Benedict’s method	Reducing substances in blood and urine
Alkaline Ferric Reduction method (Hagedorn-Jensen)	Ferricyanide ---(Glucose)--> Ferrocyanide     (Yellow)                                     (Colorless)
Ortho-toluidine (Dubowski method)	Schiff’s base
Glucose oxidase	Measures beta-D-glucose (65%)
Mutarotase	Converts alpha-D-glucose (35%) to beta-D-glucose (65%)
NADH/NADPH	Absorbance at 340nm
Polarographic glucose oxidase	Consumption of oxygen on an oxygen-sensing electrode O2 consumption α glucose concentration
Hexokinase method	Most specific method Reference method Uses G-6-PD
G-6-PD	Most specific enzyme rgt for glucose testing
Interfering substances (Glucose oxidase)	False-decreased Bilirubin Uric acid Ascorbate
Hemolysis (>0.5 g/dL Hgb)	Major interfering substance in hexokinase method (false-decreased)
Dextrostics	Cellular strip Strip w/ glucose oxidase, peroxidase and chromogen
OGTT	Janney-Isaacson method (Single dose) = most common Exton Rose (Double dose) Drink the glucose load within 5 mins
IVGTT	For patients with gastrointestinal disorders (malabsorption) Glucose: 0.5 g/kg body weight Given w/in 3 mins 1st blood collection: after 5 mins of IV glucose
Requirements for OGTT	Ambulatory Fasting: 8-14 hours Unrestricted diet of 150g CHO/day for 3 days Do not smoke or drink alcohol
Glucose load	75 g = adult (WHO std) 100 g = pregnant 1.75 g glucose/kg BW = children
HbA1c	2-3 months Glucose = beta-chain of HbA1 1% increase in HbA1c = 35 mg/dL increase in plasma glucose 18-20% = prolonged hyperglycemia 7% = cutoff Specimen: EDTA whole blood Test: Affinity chromatography (preferred)
IDA and older RBCs	High HbA1c
RBC lifespan disorders	Low HbA1c
Fructosamine (Glycosylated albumin/ plasma protein ketoamine)	2-3 weeks Useful for patients w/ hemolytic anemias and Hgb variants Not used in cases of low albumin Specimen: Serum
Galactosemia	Congenital deficiency of 1 of 3 enzymes in galactose metabolism Galactose-1-phosphate uridyl transferase (most common) Galactokinase Uridine diphosphate galactose-4-epimerase
Essential fructosuria	Autosomal recessive Fructokinase deficiency
Hereditary fructose intolerance	Defective fructose-1,6-biphosphate aldolase B activity
Fructose-1,6-biphosphate deficiency	Failure of hepatic glucose generation by gluconeogenic precursors such as lactate and glycerol
Glycogen Storage Disease	Autosomal recessive Defective glycogen metabolism Test: IVGTT (Type I GSD)
Ia = Von Gierke	Glucose-6-Phosphatase deficiency (most common worldwide)
II = Pompe	Alpha-1,4-glucosidase deficiency (most common in the Philippines)
III = Cori Forbes	Debrancher enzyme deficiency
IV = Andersen	Brancher enzyme deficiency
V = McArdle	Muscle phosphorylase deficiency
VI = Hers	Liver phosphorylase deficiency
VII = Tarui	Phosphofructokinase deficiency
XII = Fanconi-Bickel	Glucose transporter 2 deficiency
CSF glucose	Collect blood glucose at least 60 mins (to 2 hrs) before the lumbar puncture (Because of the lag in CSF glucose equilibrium time)
< 0.5	Normal CSF : serum glucose ratio
C-peptide	Formed during conversion of pro-insulin to insulin
5:1 to 15:1	Normal C-peptide : insulin ratio
D-xylose absorption test	Differentiate pancreatic insufficiency from malabsorption (low blood or urine xylose)
Gerhardt’s ferric chloride test	Acetoacetate
Nitroprusside test	10x more sensitive to acetoacetate than to acetone
Acetest tablets	Acetoacetate and acetone
Ketostix	Detects acetoacetate better than acetone
KetoSite assay	Detects beta-hydroxybutyrate but not widely used
Normal Values (Carbohydrates)	RBS = <140 mg/dL FBS = 70-100 mg/dL HbA1c = 3-6% Fructosamine = 205-285 μmol/L 2-Hr PPBS = <140 mg/dL GTT: 30 mins = 30-60 mg/dL above fasting 1-Hr = 20-50 mg/dL above fasting 2-Hr = 5-15 mg/dL above fasting 3-Hr = fasting level or below
Lipids
Phospholipids	Most abundant lipid Amphipathic: polar (hydrophilic head) and nonpolar (hydrophobic side chain)
Sphingomyelin	Reference material during 3rd trimester of pregnancy Concentration is constant as opposed to lecithin Not derived from glycerol but from sphingosine (amino alcohol)
Forms of phospholipids	70% Lecithin/Phosphatidyl choline 20% Sphingomyelin 10% Cephalin
TLC + Densitometric quantitation	Method for L/S ratio
Microviscosity	Measured by fluorescence polarization
Cholesterol	Not a source of fuel Not affected by fasting 70% Cholesterol ester (plasma/serum) 30% Free cholesterol (plasma/serum and RBC)
LCAT	Esterification of cholesterol
Apo A-1	Activator of LCAT
Cholesterol increases after the age of 50	2 mg/dL/year between 50 and 60 years old
Liebermann Burchardt	Cholestadienyl Monosulfonic acid Green end color
Salkowski	Cholestadienyl Disulfonic acid Red end color
Color developer mixture (Cholesterol)	Glacial acetic acid Acetic anhydride Conc. H2SO4
One-step method	Colorimetry (Pearson, Stern and Mac Gavack)
Two-step method	Color. + Extraction (Bloor’s)
Three-step method	Color. + Extract. + Saponification (Abell-Kendall)
Four-step method	Color. +Extract. + Sapon. + Precipitation (Schaenheimer Sperry, Parekh and Jung)
Abell, Levy and Brodie mtd (Chemical method)	CDC reference method for cholesterol: -Hydrolysis/saponification (Alc. KOH) -Hexane extraction -Colorimetry (Liebermann-Burchardt)
Triglycerides	Most insoluble lipid Main storage lipid in man (adipose tissue) – 95% Fasting: 12 hours
Triglyceride increases after the age of 50	2 mg/dL/year between 50 and 60 years old
Van Handel & Zilversmith (Colorimetric)	Chromotropic acid (+) Blue color compound
Hantzsch Condensation (Fluorometric)	Diacetyl acetone (+) Diacetyl lutidine compound
Modified Van Handel and Zilversmith (Chemical method)	CDC reference method for triglycerides: -Alkaline hydrolysis -Chloroform extraction à extract treated w/ silicic acid -Color reaction w/ chromotropic acid – meas. HCHO (+) Pink colored
Fatty acids	Short chain = 4-6 C atoms Medium chain = 8-12 C atoms Long chain = >12 C atoms Saturated = w/o double bonds Unsaturated = w/ double bonds Substrate for gluconeogenesis Most is bound to albumin
Palmitic acid	16:0
Stearic acid	18:0
Oleic acid	18:1
Linoleic acid	18:2
Arachidonic acid	20:4
Lipoprotein lipase (Lipemia clearing factor)	Hydrolyzes TAG in lipoproteins, releasing fatty acid and glycerol
Hepatic lipase	Hydrolyzes TAG and phospholipids from HDL Hydrolyzes lipids on VLDL and IDL
Endothelial lipase	Hydrolyzes phospholipids and TAG in HDL
Apolipoprotein	Protein component of lipoprotein Amphipathic helix – ability of proteins to bind to lipids
Chylomicrons	Largest and least dense Produced by the intestine SG: <0.95 80-95% TAG (exogenous) Apo B-48 (Major) EP: Origin
VLDL	Secreted by the liver SG: 0.95-1.006 65% TAG (endogenous) Apo B-100 (Major) EP: pre-beta
LDL	Synthesized by the liver SG: 1.006-1.063 50% CE Apo B-100 (Major) EP: beta Cholesterol transport: LiveràTissues Target of cholesterol lowering therapy Better marker for CHD risk
HDL	Smallest but dense SG: 1.063-1.21 45-55% protein 26-32% phospholipid Apo A-1 (Major) EP: alpha Produced by the liver and intestine Reverse cholesterol transport: TissueàLiver
IDL	Product of VLDL catabolism Seen in Type 3 hyperlipoproteinemia (Apo E-III def.; beta-VLDL) SG: 1.006-1.019
Lp(a)	Sinking pre-beta lipoprotein SG: 1.045-1.080 Apo B-100 EP: pre-beta (VLDL) UC: like LDL Independent risk factor for atherosclerosis
LpX	Found in obstructive jaundice (cholestasis) and LCAT deficiency 90% FC and PL Apo C and albumin
Beta-VLDL	Floating beta-lipoprotein SG: <1.006 EP: beta (LDL) UC: like VLDL Found in type 3 hyperlipoproteinemia (Apo E-III def; IDL) Rich in cholesterol content than VLDL
Lipoprotein methodologies	Specimen: sample from serum separator tubes (preferred) EDTA plasma: choice for research studies of LPP fractions Fasting state: TAG à VLDL Nonfasting state: TAG à CM
Ultracentrifugation	Reference method for LPP quantitation Reagent: Potassium bromide (SG: 1.063) Ultracentrifugation of plasma for 24 hours Expressed in Svedberg units
Electrophoresis	Electrophoretic pattern: (+) HDL ßVLDL ß LDL ß CM (Origin) (-) Agarose gel: sensitive medium VLDL: migrates w/ alpha2-globulin (pre-beta)
Chemical precipitation	Uses polyanions (heparin and divalent cations) and polyethylene glycol Dextran sulfate-Mg2+ Heparin-Mn2+
3-step procedure: Ultracentrifugation Precipitation Abell-Kendall assay	CDC Reference method for HDL
Beta quantification + Ultracentrifugation + Chemical precipitation	Method for LDL Sample: EDTA plasma
Immunoturbidimetric assay	Measures Lipoprotein (a)
LDL Cholesterol	Total Cholesterol – HDL – VLDL
Friedewald method	Most commonly used VLDL = TAG/2.175 (mmol/L) VLDL = TAG/5 (mg/dL) Not applicable if TAG is >400 mg/dL
De Long method	VLDL = TAG/2.825 (mmol/L) VDL = TAG/6.5 (mg/dL)
Apo A-1	Activates LCAT
Apo B-100	LDL à LDL receptor
Apo B-48	CM (major) Not recognized by LDL receptor
Apo C-II	Activates LPL
Apo D	Activates LCAT
Apo E	Apo E-4: associated w/ high LDL, higher risk of CHD and Alzheimer’s disease
Apo(a)	Lp(a) Homologous to plasminogen
Abetalipoproteinemia (Basses-Kornzweig syn.)	Autosomal recessive Defective apo B synthesis Deficient fat soluble vitamins
Niemann-Pick disease	Sphingomyelinase deficiency
Tangier’s disease	Deficiency of HDL (1-2 mg/dL) Defects in the gene for the ABCA1 transporter
LPL deficiency (Chylomicronemia)	TAG = 10,000 mg/dL Do not develop premature coronary disease (CM are not atherogenic) Abdominal pain and pancreatitis
LCAT deficiency	Fish-eye disease Low HDL
Tay-Sachs disease	Hexosaminidase A deficiency
Fredrickson Classification
Type 1	LPL deficiency (Chylomicronemia) Increased: CM (TAG)
Type 2a	Familial hypercholesterolemia Increased: LDL (cholesterol)
Type 2b	Combined hyperlipidemia (most common primary hyperlipidemia) Increased: LDL (cholesterol), VLDL (TAG)
Type 3	Dysbetalipoproteinemia Increased: IDL, (+) beta-VLDL (+) Apo E-II (+)  Eruptive and palmar xanthomas
Type 4	Hypertriglyceridemia Increased: VLDL (TAG)
Type 5	Increased: VLDL (Endo.TAG), CM (Exo.TAG)
Normal Values (Lipids)	Cholesterol: Desirable = <200 mg/dL Borderline high = 200-239 mg/dL High = >240 mg/dL Triglycerides: Desirable = <150 mg/dL Borderline high = 150-199 mg/dL High = 200-499 Very high = >500 mg/dL HDL: Low = <40 mg/dL (Cutoff) High = >60 mg/dL LDL: Optimal = <100 mg/dL Near/above optimal = 100-129 mg/dL Borderline high = 130-159 mg/dL High = 160-189 mg/dL Very high = >190 mg/dL
Proteins
Proteis	First rank of importance
Proteins	Amphoteric: positive and negative charges Effective blood buffers Synthesized by the liver except immunoglobulins (plasma cells) Provide 12-20% of total daily body energy requirement Composed of 50-70% of the cell’s dry weight
Primary structure	Amino acid sequence Det. the identity of protein, molecular structure, function binding capacity, recognition ability
Secondary structure	Winding of polypeptide chain Specific 3-D conformations: alpha-helix, beta-pleated sheath, bend form
Tertiary structure	Actual 3-D configuration Folding pattern Physical and chemical properties of proteins
Quarternary structure	Association of 2 or more polypeptide chains à protein
Albumin	No quarternary structure
Glucogenic amino acids	Alanine (pyruvate) Arginine (alpha-ketoglutarate) Aspartate (oxaloacetate)
Ketogenic amino acids	Degraded to acetyl-CoA Leucine Lysine
Simple proteins	Hydrolysis à Amino acids Fibrous: fibrinogen, troponins, collagen Globular: hemoglobin, plasma proteins, enzymes, peptide hormones
Conjugated proteins	Protein (apoprotein) + nonprotein moiety (prosthetic group) Metalloproteins: ferritin, ceruloplasmin, hemoglobin, flavoproteins Lipoproteins: VLDL, HDL, LDL, CM Glycoproteins: haptoglobin, alpha1-antitrypsin (10-40% CHO) Mucoproteins or proteoglycans: Mucin (CHO > CHON) Nucleoproteins: Chromatin (combined w/ nucleic acids)
Nitrogen balance	Balance bet. anabolism and catabolism
Negative nitrogen balance	Catabolism > anabolism Excessive tissue destruction
Positive nitrogen balance	Anabolism > catabolism Growth and repair processes
Prealbumin (Transthyretin)	Transports thyroxine and retinol (Vit. A) Landmark to confirm that the specimen is really CSF
Albumin	Maintains osmotic pressure Negative acute phase reactant
Alpha1-antitrypsin	Acute phase reactant Major inhibitor of protease activity 90% of alpha1-globulin band
Alpha1-fetoprotein	Gestational marker Tumor marker: hepatic and gonodal cancers Screening test for fetal conditions (Spx: maternal serum) Amniotic fluid: confirmatory test Increased: Hepatoma, spina bifida, neural tube defects Decreased: Down Syndrome (Trisomy 21)
Alpha1-acid glycoprotein/ orosomucoid	Low pI (2.7) Negatively charged even in acid solution
Alpha1-antichymotrypsin	Acute phase reactant Binds and inactivates PSA Increased: Alzheimer’s disease, AMI, infection, malignancy, burns
Haptoglobin (alpha2)	Acute phase reactant Binds free hemoglobin (alpha chain)
Ceruloplasmin (alpha2)	Copper binding (6-8 atoms of copper are attached to it) Has enzymatic activities Decreased: Wilson’s disease (copper à skin, liver, brain, cornea [Kayser-Fleisher rings])
Alpha2-macroglobulin	Larges major nonimmunoglobulin protein Increased: Nephrotic syndrome (10x) Forms a complex w/ PSA
Group-specific component (Gc)-globulin (bet. alpha1 and alpha2)	Affinity w/ vitamin D and actin
Hemopexin (beta)	Binds free heme
Beta2-microglobulin	HLA Filtered by glomeruli but reabsorbed
Transferrin/Siderophilin (beta)	Negative acute phase reactant Major component of beta2-globulin fraction Pseudoparaproteinemia in severe IDA Increased: Hemochromatosis (bronze-skin), IDA
Complement (beta)	C3: major
Fibrinogen (bet. beta and gamma)	Acute phase reactant Between beta and gamma globulins
CRP (gamma)	General scavenger molecule Undetectable in healthy individuals hsCRP: warning test to persons at risk of CAD
Immunoglobulins (gamma)	Synthesized by the plasma cells IgG>IgA>IgM>IgD>IgE
Myoglobin	Marker: Ischemic muscle cells, chest pain (angina), AMI
Troponins	Most important marker for AMI
TnT (Tropomyosin-binding subunit)	Specific for heart muscle Det. unstable angina (angina at rest)
TnI (Inhibitory subunit or Actin-binding unit)	Only found in the myocardium Greater cardiac specificity than TnT Highly specific for AMI 13x more abundant in the myocardium than CK-MB Very sensitive indicator of even minor amount of cardiac necrosis
TnC	Binds calcium ions and regulate muscle contractions
Glomerular proteinuria	Most common and serious type Often called albuminuria
Tubular proteinuria	Defective reabsorption Slightly increased albumin excretion
Overload proteinuria	Hemoglobinuria Myoglobinuria Bence-Jones proteinuria
Postrenal proteinuria	Urinary tract infection, bleeding, malignancy
Microalbuminuria	Type 1 DM Albumin excretion ≥30 mg/g creatinine (cutoff: DM) but ≤300 mg/g creatinine Microalbuminuria: 2 out of 3 specimens submitted are w/ abnormal findings (w/in 6 months)
CSF Oligoclonal banding	2 or more IgG bands in the gamma region: Multiple sclerosis Encephalitis Neurosyphilis Guillain-Barre syndrome Neoplastic disorders
Serum Oligoclonal banding	Leukemia Lymphoma Viral infections
Alkaptonuria	Ochronosis (tissue pigmentation)
Homocystinuria	Impaired activity of cystathione beta-synthetase Elevated homocysteine and methionine in blood and urine Screen: Modified Guthrie test (Antagonist: L-methionine sulfoximine)
MSUD	Markedly reduced or absence of alpha-ketoacid decarboxylase 4 mg/dL of leucine is indicative of MSUD Screen: Modified Guthrie test (Antagonist: 4-azaleucine) Diagnostic: Amino acid analysis (HPLC)
PKU	Deficiency of tetrahydrobiopterin (BH4) à elevated blood phenylalanine
Normal Values (Proteins)	Total protein = 6.5-8.3 g/dL Albumin = 3.5-5.0 g/dL Globulin = 2.3-3.5 g/dL