NEET MDS Lessons
Biochemistry
Protein electrophoresis is a laboratory technique used to separate proteins based on their size, charge, or other physical properties. It plays a vital role in diagnosing and monitoring various diseases, especially those involving abnormal protein production or structure.
Types of Protein Electrophoresis
1. SPE (Serum Protein Electrophoresis)
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Principle: Separation of serum proteins based on their charge.
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Major Fractions:
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Albumin
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Alpha-1 globulin
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Alpha-2 globulin
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Beta globulin
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Gamma globulin
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Clinical Applications:
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Multiple Myeloma: Detects monoclonal spike (M-protein) in gamma region.
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Chronic Infections: Polyclonal increase in gamma globulins.
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Nephrotic Syndrome: Decreased albumin, increased alpha-2 globulin.
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Liver Disease: Altered albumin and beta-gamma bridging.
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2. Hemoglobin Electrophoresis
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Principle: Separation of hemoglobin variants based on charge differences.
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Common Hemoglobins:
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HbA, HbA₂, HbF, HbS, HbC, HbE
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Clinical Applications:
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Thalassemia: Elevated HbA₂ or HbF levels.
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Sickle Cell Disease: Presence of HbS.
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Hemoglobinopathies: Differentiates variants like HbC, HbE, etc.
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3. SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis)
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Principle: Proteins are denatured and separated by molecular weight.
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SDS: Imparts uniform negative charge, eliminating charge-based separation.
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Clinical Applications:
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Protein Purification: Identifies specific proteins in research and diagnostics.
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Genetic Disorders: Detects abnormal or truncated proteins.
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Muscular Dystrophies: Analyzes dystrophin protein expression.
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Cancer Research: Studies tumor markers and oncogenic proteins.
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Sphingosine is an amino alcohol present in sphingomyelins (sphingophospholipids). They do not contain glycerol at all.
Sphingosine is attached by an amide linkage to a fatty acid to produce ceramide. The alcohol group of sphingosine is bound to phosphorylcholine in sphingomyelin structure. .
Sphingomyelins are important constituents of myelin and are found in good quantity in brain and nervous tissues.
- There are two important phospholipids, Phosphatidylcholine and Phosphatidylserine found the cell membrane without which cell cannot function normally.
- Phospholipids are also important for optimal brain health as they found the cell membrane of brain cells also which help them to communicate and influence the receptors function. That is the reason food stuff which is rich in phospholipids like soy, eggs and the brain tissue of animals are good for healthy and smart brain.
- Phospholipids are the main component of cell membrane or plasma membrane. The bilayer of phospholipid molecules determine the transition of minerals, nutrients, and drugs in and out of the cell and affect various functions of them.
- As phospholipids are main component of all cell membrane, they influence a number of organs and tissues, such as the heart, blood cells and the immune system. As we grown up the amount of phospholipids decreases and reaches to decline.
- Phospholipids present in cell membrane provide cell permeability and flexibility with various substances as well its ability to move fluently. The arrangement of phospholipid molecules in lipid bilayer prevent amino acids, carbohydrates, nucleic acids, and proteins from moving across the membrane by diffusion. The lipid bi-layer is usually help to prevent adjacent molecules from sticking to each other.
- The selectivity of cell membrane form certain substances are due to the presence of hydrophobic and hydrophilic part molecules and their arrangement in bilayer. This bilayer is also maintained the normal pH of cell to keeps it functioning properly.
- Phospholipids are also useful in the treatment of memory problem associated with chronic substances as they improve the ability of organism to adapt the chronic stress.
TRIGLYCEROL
Triacylglycerols (formerly triglycerides) are the esters of glycerol with fatty acids. The fats and oils that are widely distributed in both plants and animals are chemically triacylglycerols.
They are insoluble in water and non-polar in character and commonly known as neutral fats.
Triacylglycerols are the most abundant dietary lipids. They are the form in which we store reduced carbon for energy. Each triacylglycerol has a glycerol backbone to which are esterified 3 fatty acids. Most triacylglycerols are "mixed." The three fatty acids differ in chain length and number of double bonds
Structures of acylglycerols :
Monoacylglycerols, diacylglycerols and triacylglycerols, respectively consisting of one, two and three molecules of fatty acids esterified to
a molecule of glycerol
Lipases hydrolyze triacylglycerols, releasing one fatty acid at a time, producing diacylglycerols, and eventually glycerol
Glycerol arising from hydrolysis of triacylglycerols is converted to the Glycolysis intermediate dihydroxyacetone phosphate, by reactions catalyzed by:
(1) Glycerol Kinase
(2) Glycerol Phosphate Dehydrogenase
Free fatty acids, which in solution have detergent properties, are transported in the blood bound to albumin, a serum protein produced by the liver.
Several proteins have been identified that facilitate transport of long chain fatty acids into cells, including the plasma membrane protein CD36
Anaerobic organisms lack a respiratory chain. They must reoxidize NADH produced in Glycolysis through some other reaction, because NAD+ is needed for the Glyceraldehyde-3-phosphate Dehydrogenase reaction (see above). Usually NADH is reoxidized as pyruvate is converted to a more reduced compound, that may be excreted.
The complete pathway, including Glycolysis and the re-oxidation of NADH, is called fermentation.
For example, Lactate Dehydrogenase catalyzes reduction of the keto group in pyruvate to a hydroxyl, yielding lactate, as NADH is oxidized to NAD+.
Skeletal muscles ferment glucose to lactate during exercise, when aerobic metabolism cannot keep up with energy needs. Lactate released to the blood may be taken up by other tissues, or by muscle after exercise, and converted via the reversible Lactate Dehydrogenase back to pyruvate
Fermentation Pathway, from glucose to lactate (omitting H+):
glucose + 2 ADP + 2 Pi → 2 lactate + 2 ATP
Anaerobic catabolism of glucose yields only 2 “high energy” bonds of ATP.
The Bicarbonate Buffer System
This is the main extracellular buffer system which (also) provides a means for the necessary removal of the CO2 produced by tissue metabolism. The bicarbonate buffer system is the main buffer in blood plasma and consists of carbonic acid as proton donor and bicarbonate as proton acceptor :
H2CO3 = H+ + HCO3–
If there is a change in the ratio in favour of H2CO3, acidosis results.
This change can result from a decrease in [HCO3 − ] or from an increase in [H2CO3 ]
Most common forms of acidosis are metabolic or respiratory
Metabolic acidosis is caused by a decrease in [HCO3 − ] and occurs, for example, in uncontrolled diabetes with ketosis or as a result of starvation.
Respiratory acidosis is brought about when there is an obstruction to respiration (emphysema, asthma or pneumonia) or depression of respiration (toxic doses of morphine or other respiratory depressants)
Alkalosis results when [HCO3 − ] becomes favoured in the bicarbonate/carbonic acid ratio
Metabolic alkalosis occurs when the HCO3 − fraction increases with little or no concomitant change in H2CO3
Severe vomiting (loss of H+ as HCl) or ingestion of excessive amounts of sodium bicarbonate (bicarbonate of soda) can produce this condition
Respiratory alkalosis is induced by hyperventilation because an excessive removal of CO2 from the blood results in a decrease in [H2CO3 ]
Alkalosis can produce convulsive seizures in children and tetany, hysteria, prolonged hot baths or lack of O2 as high altitudes.
The pH of blood is maintained at 7.4 when the buffer ratio [HCO3 − ] / [ H2CO3] becomes 20
Parathyroid Hormone
Parathyroid hormone (PTH), parathormone or parathyrin, is secreted by the chief cells of the parathyroid glands.
It acts to increase the concentration of calcium (Ca2+) in the blood, whereas calcitonin (a hormone produced by the parafollicular cells of the thyroid gland) acts to decrease calcium concentration.
PTH acts to increase the concentration of calcium in the blood by acting upon the parathyroid hormone 1 receptor (high levels in bone and kidney) and the parathyroid hormone 2 receptor (high levels in the central nervous system, pancreas, testis, and placenta).
Effect of parathyroid hormone in regulation of serum calcium.
Bone -> PTH enhances the release of calcium from the large reservoir contained in the bones. Bone resorption is the normal destruction of bone by osteoclasts, which are indirectly stimulated by PTH forming new osteoclasts, which ultimately enhances bone resorption.
Kidney -> PTH enhances active reabsorption of calcium and magnesium from distal tubules of kidney. As bone is degraded, both calcium and phosphate are released. It also decreases the reabsorption of phosphate, with a net loss in plasma phosphate concentration. When the calcium:phosphate ratio increases, more calcium is free in the circulation.
Intestine -> PTH enhances the absorption of calcium in the intestine by increasing the production of activated vitamin D. Vitamin D activation occurs in the kidney. PTH converts vitamin D to its active form (1,25-dihydroxy vitamin D). This activated form of vitamin D increases the absorption of calcium (as Ca2+ ions) by the intestine via calbindin.