1 - KIDNEY AND LIVER FUNCTION TESTS

Blood Urea Nitrogen

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

Major end product (45%) of protein and amino acid catabolism

1st to be elevated in kidney injury

10-20:1

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

Blood Urea Nitrogen

BUN: Creatinine ratio

Creatinine

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

End product of muscle metabolism derived from creatine

Generation remains fairly constant

Most specific marker in kidney injury

Blood Uric Acid

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

Major product of purine catabolism

Final breakdown of nucleic acid catabolism in humans

Monosodium Urate

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

Uric acid in crystals

Tophi

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

Uric acid in joints + inflammation

Diacetyl Monoxime Method

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Chemical Method (1)

Yellow

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Diacetyl Monoxime Method

Color of end product

Coupled Urease/ Glutamate Dehydrogenase Method – UV Enzymatic Method

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Enzymatic Method (1)

Urease Method

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Enzymatic Method (2)

Urease Method

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Enzymatic Method (2)

Most commonly used method

Derived from jack beans

Isotope Dilution Mass Spectrometry

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Another methodology

Jaffe Method

Kidney Function Tests

Methodologies

Creatinine

1st methodology

Yellow

Saturated picric acid

10% NaOH

Alkaline picrate solution

Kidney Function Tests

Methodologies

Creatinine

Jaffe Method

Reagent: (color)

A:

B:

A + B:

Red orange complex/red tautomer

Kidney Function Tests

Methodologies

Creatinine

Jaffe Method

APS + Serum =

Lloyd’s Reagent

Fuller’s Earth

Kidney Function Tests

Methodologies

Creatinine

Modified Jaffe Method Reagents

_: Na Al Si
_: Al Mg Silicate

Creatinine Aminohydrolase – CK Method

Creatininase Hydrogen Peroxide Method

Creatininase-Creatine Aminohydrase

Kidney Function Tests

Methodologies

Creatinine

Enzymatic Method (3)

Reduction-Oxidation React

Caraway Method

Kidney Function Tests

Methodologies

Blood Uric Acid

Chemical Method (2)

Uricase Method

Kidney Function Tests

Methodologies

Blood Uric Acid

Enzymatic Method (most common)

Isotope dilution mass spectrometry

Kidney Function Tests

Methodologies

Blood Uric Acid

Last method

Azotemia

Kidney Function Tests

Disease Correlation

Elevated level of nitrogenous substances like urea and creatinine in the blood

Pre-renal Azotemia

Hypoperfusion

Kidney Function Tests

Disease Correlation

Cause: _

Dehydration, shock, CHF

Increased: BUN

Normal: Creatinine

Renal Azotemia

Tissue damage

Kidney Function Tests

Disease Correlation

Cause: _

Decreased GFR

Striking BUN level but slowly rising creatinine value

>100 mg/dL

>20 mg/dL

>12 mg/dL

Kidney Function Tests

Disease Correlation

Renal Azotemia

BUN = _

Creatinine = _

Uric acid = _

Postrenal Azotemia

Obstruction

Kidney Function Tests

Disease Correlation

Cause: _

Decreased GFR

Nephrolithiasis, cancer or tumors of GUT

Creatinine = normal or slightly increased

Uremia

Kidney Function Tests

Disease Correlation

Clinical syndrome comprised of marked elevation in plasma urea and other nitrogenous waste products, accompanied by acidemia and electrolyte imbalance of renal failure

Osmolality

Kidney Function Tests

An expression of concentration in terms of the total number of solute particles per kilogram of solvent

Freezing point

Kidney Function Tests

Principle of osmolality

colligative property

Kidney Function Tests

Osmolality

Measured using the _ _ of sample

280-295

300-900

50-1200

Kidney Function Tests

Osmolality

Reference range:

Serum: _-_ mOsm/kg

24-hour urine: _-_ mOsm/kg

Random urine: _-_ mOsm/kg

Kidney injury molecule-1 (KIM-1)

Neutrophil gelatinase associated lipocalin (NGAL)

Insulin-like GF binding protein 7 (IGFB7)

Tissue inhibitor metalloproteinase-2 (TIMP-2)

Kidney Function Tests

Biomarkers of Kidney Injury (4)

Synthetic function

Conjugation

Detoxification & drug metabolism

Excretory and secretory

Storage

Liver Function Tests

Liver Functions (5)

Primary Structure

Proteins

Protein Structures

Amino acid sequence

Secondary Structure

Proteins

Protein Structures

Winding of polypeptide chain

Specific 3-D conformations: alpha-helix, beta-pleated sheath, bend form

alpha-helix, beta-pleated sheath, bend form

Proteins

Protein Structures

Secondary Structure

Specific 3-D conformations: (3)

Tertiary Structure

Proteins

Protein Structures

Actual 3-D configuration

Ex: Myoglobin

Quaternary Structure

Proteins

Protein Structures

Association of 2 or more polypeptide chains = protein

Ex: Hemoglobin

Simple Proteins

Proteins

Types of Proteins

Hydrolysis will yield amino acids

Fibrous

Globular

Proteins

Types of Proteins

Types of simple proteins (2)

fibrinogen, troponin

Proteins

Types of Proteins

Simple Proteins

Examples of Fibrous

hemoglobin, plasma proteins

Proteins

Types of Proteins

Simple Proteins

Examples of Globular

Conjugated Proteins

Proteins

Types of Proteins

Protein (apoprotein) + nonprotein moiety (prosthetic group)

Metalloproteins

Lipoproteins

Glycoproteins

Mucoproteins

Nucleoproteins

Proteins

Types of Proteins

Types of conjugated proteins (5)

ferritin, ceruloplasmin

Proteins

Types of Proteins

Simple Proteins

Examples of Metalloproteins

CM, VLDL, LDL, HDL

Proteins

Types of Proteins

Simple Proteins

Examples of Lipoproteins

haptoglobin, AAT

Proteins

Types of Proteins

Simple Proteins

Examples of Glycoproteins

mucin

Proteins

Types of Proteins

Simple Proteins

Examples of Mucoproteins

chromatin

Proteins

Types of Proteins

Simple Proteins

Examples of Nucleoproteins

Kjeldahl (Digestion)

Proteins

Total Proteins

Reference method

Measurement of nitrogen content

6.54

15.1-16.8%

Proteins

Total Proteins

Kjeldahl (Digestion)

1 g N2 = _ grams of protein

_ = nitrogen content of CHON

Sulfuric acid

Proteins

Total Proteins

Kjeldahl (Digestion)

Reagent

Ammonia

Proteins

Total Proteins

Kjeldahl (Digestion)

End product

Biuret

Proteins

Total Proteins

Most widely used (IFCC)

Requires at least 2 peptide bonds

Alkaline CuSO4; Rochelle salt (NaK Tartrate); NaOH; KI

Proteins

Total Proteins

Biuret

Reagent (4)

Violet (545 nm)

Proteins

Total Proteins

Biuret

End product

Folin-Ciocalteu (Lowry)

Proteins

Total Proteins

Highest analytical sensitivity

Oxidation of phenolic compounds

Phenol; Biuret

Proteins

Total Proteins

Folin-Ciocalteu (Lowry)

Reagent (2)

Blue color

Proteins

Total Proteins

Folin-Cioicalteu

End product

280

210

tryptophan, tyrosine and phenylalanine

Proteins

Total Proteins

UV Absorption Method

Proteins absorb light at _ nm and _ nm

280 nm = (3)

Refractometry

Proteins

Total Proteins

Alternative test

Measurement of RI of solutes in serum

Turbidimetry

Proteins

Total Proteins

Formation of a uniform fine precipitate which blocks light

SSA or TCA

Proteins

Total Proteins

Turbidimetry

Reagent

Nephelometric Methods

Proteins

Total Proteins

Formation of a uniform fine precipitate which scatters light

SSA or TCA

Proteins

Total Proteins

Nephelometric Methods

Reagent

Salt fractionation

Proteins

Total Proteins

Globulins are separated from albumin by salting-out procedures using sodium salts

Sodium sulfate salt

Proteins

Total Proteins

Salt fractionation

Reagent

Prothrombin Time (Vitamin K Response Test)

Proteins

Differentiates intrahepatic from extrahepatic liver disease

II, VII, IX, X, Protein C, Protein S

Proteins

Vitamin K dependent factors

Hydroquinone

Proteins

other name for Vitamin K

SPE

SPE

Cellulose acetate

Serum Protein Electrophoresis

Medium used

Prealbumin

Serum Protein Electrophoresis

Also called as transthyretin which carries thyroxine and retinol

Abundant in CSF

All have “a1” in the name

Serum Protein Electrophoresis

α1

Ceruloplasmin, haptoglobin, a2 macroglobulin

Serum Protein Electrophoresis

α2

Hemoglobin, transferrin

Serum Protein Electrophoresis

ꞵ

Immunoglobulins

IgG, IgA, IgM, IgD, IgE

Serum Protein Electrophoresis

Ɣ:

Hypoalbuminemia

Serum Protein Electrophoresis

↓ albumin

Hyperalbuminemia

Serum Protein Electrophoresis

↑ albumin

Emphysema

Serum Protein Electrophoresis

↓ α1

Nephrotic syndrome

Serum Protein Electrophoresis

↑ α2

Liver cirrhosis

Serum Protein Electrophoresis

Ꞵ + Ɣ bridging

Endosmosis

Serum Protein Electrophoresis

Proteins that migrate in the same charge

Severe dehydration

Prolonged tourniquet application

Proteins

Albumin

Hyperalbuminemia (2)

Decreased synthesis

Proteins

Albumin

Hypoalbuminemia

CLD

Malabsorption syndrome

Malnutrition and muscle wasting

Increased loss

Proteins

Albumin

Hypoalbuminemia

Nephrotic syndrome

Massive burns

Protein-losing enteropathy

Orthostatic proteinuria

Increased catabolism

Proteins

Albumin

Hypoalbuminemia

Massive burns

Widespread malignancy

Thyrotoxicosis

Analbuminemia

Proteins

Albumin

Absence or inability to synthesize albumin

Bisalbuminemia

Proteins

Albumin

Presence of two albumin bands

Bromocresol Green – most common

Bromocresol Purple – most specific

Methyl Orange

HABA (2-(4'-hydroxyazobenzene)-benzoic acid)

Proteins

Albumin

Dyes used for Albumin measurement (4)

total protein

Proteins

Albumin

Albumin/Globulin Ratio

Ratio of albumin and globulin in relation to the _ _

1.3-3:1

Proteins

Albumin

Albumin/Globulin Ratio

NV

Cirrhosis, multiple myeloma, Waldenstrom macroglobulinemia

Proteins

Albumin

Albumin/Globulin Ratio

Inverted ratio (3)

Bilirubin

Test Measuring Conjugation and Excretion Function

End product of hemoglobin metabolism and the principal pigment in bile

myoglobin

catalase

cytochrome oxidase

Test Measuring Conjugation and Excretion Function

Bilirubin

Also formed from _, _ and _ _

Unconjugated

Water-insoluble

Non-polar

Indirect acting

Hemobilirubin

Test Measuring Conjugation and Excretion Function

Bilirubin 1 (5)

Conjugated

Water-soluble

Polar

Direct acting

Cholebilirubin

Test Measuring Conjugation and Excretion Function

Bilirubin 2 (5)

B1

B1 + B2

B2

Types of Jaundice	Pre-hepatic	Hepatic	Post Hepatic
Bilirubin increased	_	_	_

Increased

±

Absent

Types of Jaundice	Pre-hepatic	Hepatic	Post Hepatic
Urine urobilinogen	_	_	_

Normal

Dark Yellow

Dark Yellow

Types of Jaundice	Pre-hepatic	Hepatic	Post Hepatic
Urine color	_	_	_

Dark brown

Normal

Clay colored

Types of Jaundice	Pre-hepatic	Hepatic	Post Hepatic
Stool color	_	_	_