Maha Shivaratri Celebration-Monday, March 7, 2016 Maha Rudra ...
Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.
-
Upload
esmond-gibson -
Category
Documents
-
view
218 -
download
4
Transcript of Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.
![Page 1: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/1.jpg)
Clinical diagnostic biochemistry - 4
Dr. Maha Al-Sedik2015
CLS 334
![Page 2: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/2.jpg)
What are Proteins?
Proteins are made of amino acids.
They contain carbon, hydrogen, oxygen (like carbohydrates
and fats).
In addition, also contain nitrogen
Each amino acid has:
• Acid group (-COOH)
• Amine group (-NH2)
• Side chain (unique)
![Page 3: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/3.jpg)
Amino Acid
![Page 4: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/4.jpg)
![Page 5: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/5.jpg)
Amino acids
Essential a.a
Non essential a.a
![Page 6: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/6.jpg)
Amino Acids
Proteins are made up of 20 AAs
o 9 are essential ( can not be formed by the body )
o 11 are nonessential :
Can be made in the body from other amino acids
![Page 7: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/7.jpg)
![Page 8: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/8.jpg)
![Page 9: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/9.jpg)
Proteins are a link of Amino acid with peptide bonds:
Dipeptides: have two amino acids bonded together.
Tripeptides: have three amino acids bonded together.
Polypeptides: have more than two amino acids bonded together.
![Page 10: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/10.jpg)
![Page 11: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/11.jpg)
Amino acid sequence: determines shape and function and what protein is made.
INSULIN
![Page 12: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/12.jpg)
Protein Denaturing
Unfolding of protein shape by heat, acids, bases or salts that
changes its ability to function.
Stomach acid untangles proteins to aid in digestion.
![Page 13: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/13.jpg)
Denaturation
![Page 14: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/14.jpg)
Protein Digestion and Absorption
Mouth
Crushed and moisten
No digestion of protein in the mouth
Stomach
protein is denatured
Enzymes start to break the peptide bonds
HCl activates pepsinogen to pepsin
Pepsin cleaves proteins into smaller polypeptides.
![Page 15: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/15.jpg)
small intestine:
In the small intestine, polypeptides broken down into tri-,
dipeptides and amino acids by pancreatic protease
(trypsin).
Cells in the small intestine absorb amino acids
to be used by the body.
![Page 16: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/16.jpg)
Amino acids in blood are filtered through the glomerular
membranes, but normally are reabsorbed in the renal tubules by
transport systems.
When the transport mechanisms become saturated or are
defective, amino acids spill into urine, resulting in a condition
known as aminoaciduria.
Aminoaciduria
Protein and the kidney:
![Page 17: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/17.jpg)
Two types of aminoaciduria have been identified:
1. Overflow aminoaciduria: occurs when the plasma concentration
of one or more amino acids exceeds the renal threshold (capacity
for reabsorption).
2. Renal aminoaciduria: occurs when plasma concentrations are
normal, but the renal tubular reabsorption system has a congenital
or acquired defect.
Aminoaciduria
![Page 18: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/18.jpg)
Causes of aminoaciduria:
Primary aminoaciduria ( overflow ):
Due to:
Inherited enzyme defect e.g. defect in the pathway of amino
acid metabolism e.g. ( cysteinuria ).
DNA abnormality.
Secondary amino aciduria ( Renal ):
Due to:
generalized renal tubular dysfunction.
![Page 19: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/19.jpg)
Many procedures are available to measure amino acids in biological
samples.
To diagnose pathological disorders, the following three groups of
tests for amino acid analysis are important:
1) Screening tests.
2) Quantitative tests.
3) Specific tests.
Analysis of amino acids
![Page 20: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/20.jpg)
Analysis of amino acidsScreening tests
Quantitative tests
Specific tests
![Page 21: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/21.jpg)
1- TLC: TLC analysis of amino acids is conducted in three stages:
(1) preparation of the sample,
(2) chromatographic separation,
(3) identification of the separated amino acids.
2- Photometric Screening Tests for Urine
Screening tests
![Page 22: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/22.jpg)
Amino acids are measured quantitatively in body fluids with a variety
of techniques, including:
1. Electrophoresis .
2. Gas chromatography (GC).
3. High-performance liquid chromatography (HPLC).
4. Ion-exchange chromatography.
Quantitative tests
![Page 23: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/23.jpg)
In addition to the general analytical techniques discussed
previously, a variety of simple tests exist that are specific for
individual amino acids.
These tests are used in the diagnosis of specific disorders.
Tests for Specific Amino Acids
![Page 24: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/24.jpg)
Plasma proteins
![Page 25: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/25.jpg)
(1) Enzymes are proteins that catalyze biochemical reactions essential
to metabolism.
(2) Hormones regulate body functions.
(3) Immunity: Antibodies and complement system protect against
infection.
(4) Osmotic pressure: Plasma proteins maintain the osmotic pressure
of plasma.
(5) They transport hormones, vitamins, metals, and drugs.
(6) Hemostasis: coagulation system.
Functions of plasma proteins:
![Page 26: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/26.jpg)
Causes of increased plasma protein levels:1. Dehydration: decreased volume of distribution due to relative
water deficiency.
2. Hemoconcentration: due to stasis of blood during prolonged
application of tourniquet.
3. Paraproteinaemia: increase in abnormal protein synthesis.
4. Hypergammaglobulinemia: increase in the antibody
concentration.
![Page 27: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/27.jpg)
Causes of decreased plasma protein levels:1. Overhydration: increased volume of distribution ( IV fluid
administration ).
2. Artifactual: blood samples from drip arm.
3. Decrease in protein synthesis: sever liver disease or
immunodiffeciency.
4. Excessive protein loss: in urine e.g. nephrotic syndrom or from
skin e.g. severe burns.
![Page 28: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/28.jpg)
o Normal total plasma protein concentration: 6 – 8.5 gm / dl.
o Electrophoresis separates serum Proteins into 5 distinct zones or
bands at barbital buffer at pH 8.6 :
Prealbumin.
Albumin.
Alpha globulin ( Alpha 1 and Alpha 2 ).
Beta globulin ( Beta 1 and Beta 2 ).
Gamma globulin: IgG, IgA, IgM, IgD, IgE and C-reactive protein.
![Page 29: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/29.jpg)
The width of each band is dependent upon the number
of proteins that are present in that fraction.
![Page 30: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/30.jpg)
Q – WHAT is the difference between serum proteins and plasma proteins in electrophoresis?
![Page 31: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/31.jpg)
Serum is a clear yellowish fluid that remains from blood plasma after
clotting factors (fibrinogen, prothrombin ect.) that have been used in
the formation of a clot.
Plasma is a clear yellowish fluid that still contains all of the clotting
factors and have not been solidified into clot.
![Page 32: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/32.jpg)
If plasma is used in electrophoresis , there will be an extra band
( fibrinogen ) between beta and gama bands.
Fibrinogen
![Page 33: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/33.jpg)
Reference: Burtis and Ashwood Saunders, Teitz fundamentals of Clinical Chemistry, 4th edition, 2000.
![Page 34: Clinical diagnostic biochemistry - 4 Dr. Maha Al-Sedik 2015 CLS 334.](https://reader036.fdocuments.in/reader036/viewer/2022062322/5697bfb71a28abf838c9f03c/html5/thumbnails/34.jpg)
THANK YOU