Blood and Urine Analysis Quiz

Practical Biochemistry Study Notes

• Course Name: Practical Biochemistry
• University: Minia University
• Faculty: Medicine
• Department: Biochemistry
• Year: First year
• Academic Year: 2024-2025

Lab Safety

• Essential Practices: Practice proper lab attire (lab coat, gloves, protective goggles, IDs), identify warning signs, understand hazard material procedures, and know how centrifuges work.
• Safe Working: Lab is a serious work environment, careless behavior is not tolerated. Safe working protects the user, other lab workers, cleaners, visitors, and the workspace.
• Safety Rules: Adhere to lab coat, book placement, and lab manual use regulations. Listen carefully to instructions and stand properly during lab sessions. Read experiment instructions beforehand.
• General Work Procedure: During lab work, ask questions, wear safety eyewear, and gloves when handling blood specimens. Avoid broken glassware and inspect glassware for cracks. Use flames cautiously and avoid carrying hot or dangerous equipment and chemicals unsupervised. Never taste or handle chemicals with your bare hands; keep lids on bottles and inform instructor about any issues.

Laboratory Instruments and Apparatuses

• Volume Measurement: Instruments for volume measurements depend on the instrument's size—a smaller diameter generally means more accurate measurement.
  • Graduated Cylinders: Used to measure various liquid volumes.
  • Syringes: Used for precise liquid transfer.
  • Burets: Instruments with stopcocks for precise liquid control.
  • Pipettes: Used to accurately measure and deliver specific portions of liquids (1mL-10mL). Automatic pipettes are most accurate (1µL-1000µL).
• Chemical Handling:
  • Test Tubes: Holding/processing liquid samples for various lab procedures (heating, dissolving, centrifugation).
  • Bottles/Containers: For storage of various chemicals and reagents. Use informative labels.
  • Pipette Pump: Used for glassware delivery without mouth suction.
  • Spatulas: Used for handling solid powdered chemicals.
  • Test Tube Racks: Hold/organize test tubes on a lab bench. Plastic racks may melt if heated excessively.
  • Test Tube Holders: Used when handling hot/unsafely handled test tubes.

Blood & Plasma & Serum

• Blood: Red-colored fluid, composed of blood cells suspended in blood plasma.
• Plasma: Straw-colored liquid component of blood containing proteins, glucose, clotting factors, minerals, and waste.
• Serum: Plasma without fibrinogen and other clotting factors. 
• Plasma Components: Water (91%), Inorganic components (Na, K, Ca, Mg, Cl, HCO3, HPO4), Organic components (plasma proteins 6-8%, hormones [cortisol, thyroxine], vitamins, lipids, blood gases (O2 and CO2), other metabolites [glucose, amino acids, urea, uric acid]). 
• Cellular Components: Blood cells (RBCs, WBCs, and platelets) comprise 40-45% of total blood volume.

Methods of Separation of Plasma and Serum

• Mixing blood with anticoagulants (heparin, oxalate, EDTA) prevents clotting. Thoroughly mix.
• Centrifuge the mixture, (2000-2500 rpm for 5-10 min). 
• Separate the supernatant fluid (plasma) or serum (plasma without clotting factors).

Protein concentration in different body fluid

• Normal plasma protein content: 6-8 g/100 mL.
• Key protein components: Albumin (3.5-5 g/dL), Globulins (2.5-3.5 g/dL), and Fibrinogen (200-400 mg/dL). 
• Most plasma proteins are synthesized in the liver (except immunoglobulins). 
• Albumin:globulin ratio (A/G ratio) is usually between 1.2:1 or 1.5:1.

CSF (Cerebrospinal Fluid)

• Normal CSF protein levels: 15-45 mg/dL.
• Infants have higher CSF protein levels than adults.
• CSF protein concentration falls rapidly from birth to six months.

Urine Analysis

• Volume: Normal adult urine volume: 1-1.8 liters/24 hours. Ratio of night-to-day urine production is usually 1:2 or 1:3.
• Color: Normal urine color: from pale yellow to deep yellow, depending on concentration. 
• Odor: Normal odor is slightly aromatic, but in standing urine, the typical odor is ammonical.
• Reaction (pH): Normal urine pH is slightly acidic (around 6.8).
• Specific Gravity: Adult values 1.001-1.025, newborn typically 1.015-1.018. A higher value typically denotes a higher concentration of solutes in the urine.
• Physical Properties:
  • Appearance  (Crystal Formation, Pus, Blood Cells, Bacteria, and Fatty substances).

Chemical Examination of Urine

• Reducing Sugars: Test positive for reducing sugars like glucose, galactose, or fructose, using Benedict's test.
• Ketone Bodies: Detection of ketone bodies using Rothera test.
• Urinary Proteins: Test for presence of urinary proteins, such as albumin, globulin, or other using heat coagulation test.
• Urea: Detect Urea utilizing the diacetyl monoxime test.
• Phosphate: presence of phosphate using ammonium molybdate test.
• Calcium: Presence/absence of calcium with ammonium oxalate test.
• Bile Salts: Test for presence of bile salts using the Hay sulfur test.

Clinical Enzymology

• Enzyme Definition: Biological catalysts accelerating reaction rates and often specific to tissues.
• Serum Enzymes: Enzymes in blood; some are essential (clotting) and others are present but not active in blood.
• Clinical Application: Measurement of tissue-specific enzyme levels can indicate injury, death, or tissue problems in organs.

DNA Extraction

• Definition: Removing DNA from cells.
• Sources: Any living tissue (plants, animals).
• DNA Extraction Procedure:
  • Crushing /Lyse: Break open cells to release DNA.
  • Extraction Buffer: Dissolve membranes, proteins, and stabilizes DNA.
  • Ice-Cold Alcohol: Precipitates DNA.
• DNA Extraction Protocol: outlines detailed steps and components, and lab equipment to conduct procedure.

PCR (Polymerase Chain Reaction)

• Definition: In vitro cloning technique amplifying specific DNA sequences efficiently.
• Principle: Repeating cycles of DNA synthesis utilizing heat-stable enzymes (and primers) for efficient target amplification.
• Components: DNA polymerase, primers, DNA template, dNTPs.
• Steps: Denaturing, Annealing, and Extending using DNA polymerase catalyzing primer extension.
• Applications: Diagnose infectious diseases, genetic disorders, genetic engineering, forensic science, and environmental studies.
• Types: Conventional, Multiplex, Nested, and other specialized PCR variants.

Urine Sheet

• This is a form to record specific urine analysis results.
• Results of urine tests are recorded, including details about physical characteristics (volume, color, odor) and chemical properties (reducing sugar, ketone bodies, proteins, urea, phosphates, and bile salts/bile pigments).

Test your knowledge on various aspects of blood and urine analysis. This quiz covers the processes and components involved in centrifugation, protein content in plasma, and indicators of health based on urine characteristics. Dive into the details of common conditions like gout and the significance of test results.