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

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

Blood Uric Acid

Kidney Function Tests

Non-Protein Nitrogenous Compounds (NPNs)

Major product of purine catabolism

Final breakdown of nucleic acid catabolism in humans

Diacetyl Monoxime Method

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Chemical Method (1)

Coupled Urease/ Glutamate Dehydrogenase Method – UV Enzymatic Method

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Enzymatic Method (1)

Isotope Dilution Mass Spectrometry

Kidney Function Tests

Methodologies

Blood Urea Nitrogen (BUN)

Another methodology

Jaffe Method

Kidney Function Tests

Methodologies

Creatinine

1st methodology

Creatinine Aminohydrolase – CK Method

Creatininase Hydrogen Peroxide Method

Kidney Function Tests

Methodologies

Creatinine

Enzymatic Method (2)

Reduction-Oxidation React

Kidney Function Tests

Methodologies

Blood Uric Acid

Chemical Method

Uricase Method

Kidney Function Tests

Methodologies

Blood Uric Acid

Enzymatic Method

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

Kidney Function Tests

Disease Correlation

Dehydration, shock, CHF

Increased: BUN

Normal: Creatinine

Renal Azotemia

Kidney Function Tests

Disease Correlation

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

Kidney Function Tests

Disease Correlation

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

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)

Synthesis and storage of amino acids, proteins

Bile drainage

Detoxification

Blood glucose regulation

Blood circulation and filtration

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

Quaternary Structure

Proteins

Protein Structures

Association of 2 or more polypeptide chains = protein

Simple Proteins

Proteins

Types of Proteins

Hydrolysis will yield amino acids

Fibrous

Globular

Proteins

Types of Proteins

Types of simple proteins (2)

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)

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

Biuret

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

SPE

SPE

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

BCG

BCP

MO

HABA

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

<1

Proteins

Albumin

Albumin/Globulin Ratio

Inverted ratio

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

Bound to albumin

Indirect acting

Hemobilirubin

Test Measuring Conjugation and Excretion Function

Bilirubin 1 (5)

Conjugated

Water-soluble

Not bound to albumin

Direct acting

Cholebilirubin

Test Measuring Conjugation and Excretion Function

Bilirubin 2 (5)

Unconjugated

Both (unconjugated and conjugated)\

Conjugated

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

Increased

Normal or decreased

Decreased or absent

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

Normal

Dark

Dark

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	_	_	_

Normal

Very high

Increased

Types of Jaundice	Pre-hepatic	Hepatic	Post Hepatic
AST and ALT	_	_	_

Normal

2-3x

10-12x

Types of Jaundice	Pre-hepatic	Hepatic	Post Hepatic
ALP	_	_	_

Gilbert’s Syndrome

Crigler-Najjar Syndrome

Dubin Johnson Syndrome

Rotor Syndrome

Lucey-Driscoll Syndrome

Derangements in Bilirubin Metabolism (5)

0.1% Sulfanilic acid + HCl

0.5% Na Nitrite

1.5% HCL

Methods of Bilirubin Assay

Diazo Reagents

Diazo A: _

Diazo B: _

Diazo Blank: _

Acidic

Red to reddish-purple

Methanol

Methods of Bilirubin Assay	pH	Color	Accelerator
Evelyn Malloy	_	_	_
Jendrassik Grof

Alkaline

Blue to blue-green

Caffeine-benzoate

Methods of Bilirubin Assay	pH	Color	Accelerator
Jendrassik Grof

Enzymes

Test for Detoxification Function

Used to assess the extent of liver damage and to differentiate hepatocellular from obstructive diseases

Ammonia

Test for Detoxification Function

Marker for liver function

Arises from deamination of amino acids

Liver

urea

kidneys

Test for Detoxification Function

Ammonia

_ removes this via portal vein circulation and converts it to _ then eliminated via the _

Hepatic coma

Test for Detoxification Function

Ammonia

Increased: _ _

19-60 ug/dL (11-35 mmol/L)

Test for Detoxification Function

Ammonia

NV:

Tobacco smoke

Test for Detoxification Function

Ammonia

Contaminant:

Direct

Test for Detoxification Function

Ammonia Methods

Nitrogen ion in PFF is converted to ammonia

Test for Detoxification Function

Ammonia Methods

Nesslerization reaction (yellow to orange brown color)

Berthelot reaction (Na nitroprusside)

Glutamate dehydrogenase

Test for Detoxification Function

Ammonia Methods

Enzymatic method