What are some preanalytical factors that might influence A.A. testing?

1. Fasting 6-8 hours prior (avoid a.a. absorbed from diet)
2. Free of Hemolysis 
3. Heparinzed Plasma 
4. Deproteinized within 30 minutes (immediately run or freeze)

What is Primary Aminoaciduria?

• Missing an enzyme essential to A.A. metabolic pathway
• Causes a buildup of toxic A.A. and/or byproducts in the blood

What is Secondary Aminoaciduria?

Renal mechanism, something wrong with renal clearence or renal absorption

Buildup of A.A with normal renal clearence 

Increased A.A. in urine 

Increased A.A in plasma

Caused by malfunction of renal reaborption of A.A 

Increased A.A in urine

Normal A.A. in plasma

What enzyme is absent in PKU?

Mousy odor in urine

Causes brain damage

Phenylalanine levels >1200umol/L

Type I Tyrosinemia

-Furmarylacetoacetate acid hydrolase (FAH) deficiency

[LETHAL!]

Absence or reduction of Alpha-ketoacid decarboxylase.

Blocks the normal metabolism of leucine, isoleucine, and valine (all essential branched chain A.A)

Which Primary Aminoaciduria has the distinct odor of sweaty feet due to a build up of isovaleric acid?

What is the enzyme deficiency?

Isovaleric Acidemia

Due to the absence of Isovaleryl-CoA dehydrogenase

What is Homocystinuria?

What does it cause a buildup of?

Absence of Cystathionine-beta-synthetase

Causes a buildup of methionine and homocystein (precursor) in the urine and plasma 

What is cystinuria?

Absence of carrier protein that helps in reabsorption of cysteine, lysine, ornithine, and arginine

Causes a build-up of amino acids in urine and renal calculi formation

1. Screening tests:

--Use Thin Layer Chromatography (stain with ninhydrin)

2.Quantitative methods: 

--Ionexchange 
--Capillary Electrophoresis 
--Tandem Mass Spec

serves as transport protein for thyroxine and triiodothyronine

rarely seen as distinct band in electrophoresis 

migrates ahead of albumin 

When is an increase of serum pre-albumin seen? Decreased?

Increased: in some cases of renal disease with GFR

Decreased: in severe liver disease, burns, salicylate poisoning, malnutrition, and inflammation

• Serves as transport protein for fatty acids, bilirubin, drugs, etc.
• maintains oncotic pressure  
• Fastest and most abundant migration on electrophoreisis

Decreased: seen in malnutrition, starvation, malabsorption, protein losing disorders, severe liver disese

Increased: dehydration and albumin therapy 

1. Alpha-1-antitrypsin (AAT)
2. Alpha-1-acid glycoprotein (AAG)
3. Alpha-1-fetoprotein (AFP)
4. Alpha-1-lipoprotein (HDL)

• a protease inhibitor
• compromises 90% of alpha1 band and migrates immediatel behind albumin

• Decrease: severe emphysema like pulmonary disease 
• Increase: infants, pregnancy, and oral contraceptive therapy  and acute inflammation

• Postive acute phase reactant 
• found in platelets, and hormone (progesterone) inactivation 

• associated with acute inflammation mostly
• also in cancer, pneumonia, pregnancy, and oral contraceptive therapy

1. Haptoglobin
2. Alpha-2-macroglobulin
3. Ceruloplasmin

in nephrotic disorders

increased serum values are found in conditions of protein loss

Wilsons Disease

Also nephrotic syndrome and advanced liver disease

1. Transferrin
2. Hemopexin
3. Betalipoprotein (LDL)
4. Complement: C3 and C4
5. Fibrinogen
6. Beta-2-Microglobulin

• Highly responsive and most sensitive acute phase reactant 
• increased during inflammation
• Used to monitor Arthrosclerosis risk

1. Kjeldahl
2. Biuret
3. Refractory 
4. UV absorption 

Dye Binding method using bromcresol green (BCG)

Intereference: Bilirubin (intereferes with albumin binding to dye)

[image]

What is this?

Nephrotic Syndrome

Name the functions of plasma proteins

1. Osmotic force/electrolyte balance 
2. involved with clotting and inflammation
3. Gammaglobulins act as antibodies 
4. maintina acid-base balance
5. source of energy

Chronic inflammation 

Chronic liver disorders

those amino acids degraded directly to acetyl coA

(Leucine and Lysine)

Potential infection (meningitis)

Neoplasmic conditions 

Multiple Sclerosis

1.Increased total protein 

2.Presence of Myelin Basic Protein (MBP)

3.Abnormal paraproteins 

4.Oligoclonal banding in CSF protein electrophoresis