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• Lection 1: Presentation in March 6, fig 37-8.
• Case studie is a debate with multiple myeloma.
• The biuret method contains Cu²⁺ in an alkaline solution and the sample would be non-hemoglobin.
• Hemolysis causes color changes and a deviation decreases lower d2, can also be caused by electrodenaturation, hep B junctions are higher.
• The presentations, lectures, and mini-quizzes are each class.
• The most proteins are synthesized by the hepatic parenchymal cells.
• A paraprotein consists of a Monoclonal Immuno.
• Enzymes assay measured using single measurements and fixed time.
• International Unit of enzyme activity is 1 micromole of substrate per minute.
• Highest CK isoenzymes of healthy is CK-MM.
• Children have higher ALP activities, ALP leaks from osteoblasts during normal growth.
• Loss of oxygen in arterial blood supply is Ischemia.
• A condition from ineffective pumping of blood is congested heart failure.
• Oxygen-binding protein of cardiac + Skeletal, myoglobin
• Creatine kinase isoenzymes "cardia isoemyma CK-2.

Exam 1 Study

• Cryoglobulins are proteins that precipitate at lower temperatures (up to 35°C).
• 3 types can be associated with or without immune complex diseases:
  • Type 1 is associated with multiple myeloma protein (25%).
  • Type 2 is a mixed multiple myeloma protein with other proteins (25%).
  • Type 3 are mixed polyclonal proteins.
• Clinical assessments:
  • Refrigerate samples for 72 hours.
  • Resuspend precipitated protein in warm saline.
  • Characterize by serum protein electrophoresis (SPE) and immunofixation electrophoresis (IFE).

Oligoclonal Bands

• Oligoclonal bands in CSF;
  • Detection of Ig in CSF of MS patients.
  • Observed in 90-95% of MS patients.
  • Indicative of Ig synthesis within the CNS.
  • Super dark blue bands
• Proteins separated by charge are amphoteric.
• Isoelectric point is where a pH where protein has net (-) charge.
  • if PH>(greater) isoelectric point, the protein has a net (-) charge, migrate towards anode (+ terminal).
  • if PH<(less) isoelectric point: protein has net (+) charge and it, migrates towards cathode (- terminal).
• Speed of migration depends largely on: the degree of ionization of the protein at the pH of the buffer system. It also depends on electric field strength, Size + shape of molecule, temp, & buffer characteristics.
• You can perform using urine or CSF, sample must be concentrated or sensitive staining methods used, identification of bence-jones.
• 1st order kinetics: rate is linearly dependent on the amount of substrate, saturation of enzyme-active sites.
• Limited # of active sites; at low conc. of substrate rate of rxn will depend on substrate conc. because there will be a large effective conc. of unfilled active Sites.
• Limited # of active sites, enzyme is constant, as substrate increases, more active Sites are filled until saturated, making it appear as the rxn rate is indep of Substrate conc.
• Competitive inhibitors: bind to the active site.
• Non-competitive inhibitors: bind enzyme at site other than the active site.
• Uncompetitive inhibitors: bind to Es complex.
  • Creatine kinase Assay (CPK,CK).
  • Creatine + ATP results phosphocreatine -ADP that can transfers a phosphate group.
  • Muscle contraction is when 2 subunits with (M+B)
• 1° Separate chromosomes are (19+14) and those are found in the cytosol of striated muscle, heart, and brain,.
• The 3 primary isoenzymes can exist. CK-BB (1) is predominant in brain, CK-MB(2) mainly in cardiac muscle, also other types of muscle, CK-MM(3) is predominant in skeletal muscle & cardiac tissue Measured by coupled rins
• Serum is the preferred heparin, EDTA, citrate.

Creatine Kinase Assay cont.

• CK-M+ (type 2 macro) is a mitochondrial isoenzyme, located between inner & outer membranes between of mitochondria
• The Macro-ck becomes an indicator of severe illness
• Macroc-ck (Type 1) is the complex: CK:CK between CK-BB +Ig) that can conjugate with lg bigger
• Type 1: most-often CK-1 complexe with IgG; sometimes CK-3 wl IgA.
  • Associated with increased mortality in gastrointestinal disorders, adenomal carcinoma, myocardial/vascular.
  • often in women after 75 minutes.
• Type 2: predominantly in adults severely ill wi malignancy or liver disease, children wi myocardial disease interferes wi assay of CK-2 by ion exchange or immunoinhibition
• Elevated in heart disease; Myocardial infarction.
• Increased Levels of: CK-2, CK-3 in Others
• Clinical significance:
  • Elevated in muscle disorders, before symptoms muscular dystrophy (esp. Duchemes usually CL-3 C(MM) Elevated in CNS disease
• "CK-2 found in cases of diseased/damaged skeletal muscle
• CK-3: acute cerebrovascular disease, cerebral ischemia, neurosurgical intervention. CK-1: head trauma.
• Thyroid disease.
• Elevated in hypothyroidism (5-50%, total: due to muscle cramping).
• mostly CK-3, some cx-2
• Decreased in hyperthyroidism
• Total CK increase assays by change in Abs340 measured (NAD/NADH)

Congestive Heart Failure.

• Edema in lungs, rest of body.
• Cause by: coronary artery disease, cardiomyopathies, myocarditis valvular disease, cardiac arrhythmias
• Inability of heart to pump blood effectively with lower cardia output.
• Left side fails: fluid accumulates in lungs, the pulmonary edema, and reduced output to systemic circulation and kidneys retain fluid.
• Right side fails: fluid accumulates in systemic venous circulation, general edema, poor blood goes to lungs.
• Decreases output to arterial circulation.
• Diagnosis:
  • Blood urea nitrogen (BUN).
    • Measure serum urea nitrogen can use either; -Enzymatic Cindirect: urease + ammonia).
      • direct (Fearon rxn). -increased assoc. wl renal failure -ICHF: ↑ BUN W/normal creatinine, Uremia the increased blood urea and Azotemia is the increase in nz-containing end products, urea, creatinine, and uric acid. -BNPs should be <100 pg/mL, unlikely 100-400 pg/mL, CHF likely and 7400 pg/mL: CHF very likely.

Enzymatic Markers for MI

• Total CK
  • CK-1 (BB) predominately in the brain.
  • CK-2 (MB): relatively specific to cart 10-20% of total CK in hearts >20% total ck in skeletal muscle
  • CK-3 (MM): cardiac + Skeletal + Elevated 4-6 hr post MI, peaks: 24hr, normal 48- 72 hr. The total must be serially measured 0-3,3-6,6-9, 12-24 hr after presentation.
• can be immuno with Ab specific for CK: and there the limits lµg/2 detect early 100% specific
• Myoglobin
• Heme-containing Oz binding protein Storage.
• Rapidly released wi muscle damage
• 1-2 hr post MI. Levels normal 10-12 hr post.
• Detected by immunoassay "ELISA, Fluroscent, turbidometric, nephelometric.
• Comparable to CK-2 (Stays elevated longer). "Not for early diagnosis 4-6 hr post, stays elevated 5-10 days +Fluorescent immunoassay.

Brain natriuretic Peptide (BNP)

Released in response to ventricle vol. expansion and pressure overload, works in tandem wi atrial natriuretic protein. Diagnosis of CHF

• <100 pg/ML, CHF unlikely
• 100-400 pg/ML CHF likely

• 400 pg/mL CHF very likely

• Provides prognostic info
• Lipoproteins are measured by km Km Constant Cenzymes
  • Michaelis-menten constant: is the substrate] when v Vmax/2 constant for a given enzyme under given conditions .

Chylomicrons.

• Lipid component: 98% (chol: 9%, tri:82%, phosp.:7% ).
• Protein component: 2%
• that comes from Intestine
• Function in transport of exogenous triglycerides.

VLDL.

• Lipid component: 92% (chol. 22%, tri: 52%, phosp:18% ).
• Protein component: 8%
• Source: Liver _Function in transport of endogenous triglycerides. IDL
• Lipid component: 85% (chol: 38%, tri: 20%, prosp: 20%). "Source: catabolism of VLDL
• Present in low conc. of fasting plasma. Precursor of LDL. LDL.
• Lipid component: 79% (chol: 47%, tri: 9%, phosp:23% ).
• Protein component: 23% (90-95% total plasma Aпов-100).
• Source: catabolism of IDL.
• Functions in cholesterol transport.

High-density lipoprotein HDL.

• HDL Lipid component: 50% (chol:19%, tri: 3%, phosp:28%). _Lecithin: important functional role in plasma cholesterol esterification is catalyzed by LCAT. "HDL Protein component: 50%
• HDL Source: Liver
• Furhter fractioned by centrifuge into HDL2 HDL3
• Triglyceride-Rich Lipoproteins Chylomicrons: 82% VLDL: 52%
• that carries VLDL High VLDL directly correlates to high triglyceride level, that transports triglycerides in the blood,
• What creamy layer on serum means?
• multiple myeloma means caused by high levels of chylomicrons.

Lipemic Specimen of High trigycerides

• that turbidity through High in VLDL+IDL Reverse Cholesterol transport macrophages with a nascent HDL - mature HDL - Liver+ Protects against atherosclerosis
• Excess-free cholesterol is removed from cells in peripheral tissues The released FC is returned can transfer of the liver for excretion in the bile
• key molecules ATP-binding cassette protein A1 (ABCAI) and Lecithin: cholesterol acyltransferase (LCAT) Scavenger receptor class B, Type 1 (SR-BI) Uptake of HDL Cholesterol. #1: cholesterol esters taken up from HDL from hepatic HDL receptors is where HDL particles are returned to circulation the cholesteral ester transferred from HDL 10 B-100-containing lipoprotein is were the cholesterol in taken up by liver by receptors for lipoprotein If HDL: ApoE can be recognized by hepatic remnant receptors because the cholesterol returned to liver is either reused or excreted

Multiple Mycloma

that can presence of paraproteins can be found in the serum (80%) and urine (50%) Bone pain, that results in anemia, fatigue causedslytic lesions, hypercalcemia, and renal insuffiency with 210 plasma cells in BM "Lipemic specimen of HDL Coronary Heart Disease LDL Risk that are high in triglycerides" VLDL The HDL: High sensitivity -CRP mg/L are high "10 not consistent w/ CHD risk <100μmol/L that are 78 high risk <100μmol/L, normal, High sensitivity - CRP mg/L and <100μmol/L. 1-2 normal <=1 low risk Formula to Calculate LDL conc 1,000-13.6 is needed is appropriate on EDTA plasma.

• "Based on ultra + polyanion centrifugation d =1.006 g/mL [VLDL] = [total] -[1.006 g/mL] (LDL) = (d=1.006 g/mL)
• (HOL
• Friedwald formula to calculate HDL conc. in plasma (mg/dL) = Total chiesterol - HOL 400mg/41

HDL-C LDL-C and CHD, LDL, and High density lipoprotein HDL

The desirable levels are the: LDL; less then 130mg 130-160, High risk is great or is above the level 40mg Low density is normal

is the High and protective LDL-C, and for normal is the:130-160

• People are develop diseases. The Type 3 Hyperlipoproteinemia with dysbetalipoproteinemia's Primary genetic defeet is to remove the that are mutants of ApoE
• that is less then 100 of this result the increase of and deposits

That test results result increased - 100km that is related the and intestinal 10% to the LDLs with no high and 100m-to-the .100km

The 1,0000-136 and the.