Untitled Quiz

Practical Biochemistry

• Course: First year, Minia University, Faculty of Medicine, Biochemistry Department
• Semester: 2024-2025
• Subject: Practical Biochemistry

Index

• Lab safety: Page 3
• Blood & Plasma & Serum: Page 11
• Electrophoresis: Page 21
• Clinical enzymology: Page 26
• Urine analysis: physical properties: Page 35
• Urine analysis: chemical properties: Page 41
• Non-protein nitrogenous compounds: Page 44
• Blood glucose & Diabetes mellitus: Page 50
• DNA extraction: Page 60
• PCR: Page 64
• Urine sheet: Page 68
• Signature: Page 70

Lab Safety

• Basic lab practices: Proper dressing (lab coat, gloves, protective goggles, IDs), identification of safety signs and hazards, elimination of hazards material, description of centrifuges and their function, and handling of lab glassware.
• Safe working practices: Importance of careful behavior, avoiding careless behavior which could endanger yourself and others in the lab, safe practices to protect yourself, other lab workers, cleaners, visitors, and your work.
• Safety rules: Instructions on appearance, attitude, placing items in cabinets, and listening to lab instructors.

General Work Procedure

• Safety precautions: Always wear approved safety googles; do not eat or drink; extreme caution with flames and hot/dangerous chemicals. Safe handling of glassware; do not taste or touch chemicals. Secure lids; report any malfunctioning equipment.
• Hygiene: Hand hygiene, keep work areas clean, return equipment, and clean spills.
• Emergency procedures: Steps of action in case of an accident with acids or alkalis, eye injury, or fainting.

Safety Practices

• Signs of hazards: List of hazard symbols: flammable, harmful/irritant, corrosive, poison/toxic, explosion, biohazard, oxidizer, environmental hazard, radioactive hazard, animal hazard, sharp instrument hazard, heat hazard, glassware hazard, chemical hazard, electrical hazard, eye & face hazard, fire hazard, laser radiation hazard, radioactive hazard, explosive hazard.

Laboratory Instruments and Apparatuses

• Instruments for Liquid Volume Measurements: Use of graduated cylinders, syringes, burets, pipettes, and volumetric flasks for accurate volume measurements.
• Pipettes: Detailed use for transferring small volumes.
• Graduated Cylinders: Used for measuring volumes of liquids
• Test tubes, Chemicals bottles: Used for chemical experiments, holding samples, etc; labels are required for accurate identification
• Pipette pump, spatula: tools used for handling chemicals; pipette pump for non-mouth suction, spatula for powdered chemicals.
• Test tube racks, holders: Used for carrying out tests, holding tubes in the lab.

Blood & Plasma & Serum

• Definitions: Blood composition, plasma (body fluid consisting of protein), and serum (plasma without clotting factors).
• Components of Blood: Plasma (water, inorganic and organic components such as proteins, hormones, vitamins, lipids, blood gases, and other metabolites), and cellular components (RBCs, WBCs, and platelets).
• Components of Serum: Same as plasma except for clotting factors.
• Separation: Methods for separating plasma and serum involves mixing blood with anticoagulants. Then centrifuge the sample to separate the components.

Protein concentration

• Plasma proteins: Normal plasma protein concentration levels (6-8 g/100 ml), list of proteins in plasma including; Albumin (3.5–5 g/dl), Globulins (2.5–3.5 g/dl), and Fibrinogen (200–400 mg/dl).
• Plasma protein synthesis location: all plasma proteins are synthesized in the liver, except Immunoglobulin.
• Albumin: Globulin ratio: Typically between 1.2:1 and 1.5:1.
• Inverted ratio: Causes include, liver and kidney diseases.

CSF (Cerebrospinal fluid), and Human breast milk composition

• CSF protein levels: Normal range (15-45 mg/dL) in adults and infants; with significant higher levels in infants.
• Human breast milk: Composition of macronutrients (fat, protein, and carbohydrate).

Protein in urine

• Normal urine protein excretion: Less than 150 mg in adult.
• Urine protein components: Albuminuria, mucoproteins (Tamm-Horsfall glycoprotein), Immunoglobulin, others such as hormones and enzymes.
• Types of albuminuria: Normo-albuminuria, Micro-albuminuria, and Macro-albuminuria.
• Causes of Proteinuria: Physiological (physical effort), Pathophysiological, Renal, Post-Renal, and Prerenal causes.
• Methods of detection: Qualitative and quantitative methods.

Enzyme Assays

• Definition: Laboratory techniques measuring enzymatic activity.
• Methods: Fluorescence, spectrophotometric, sampling, electrode, manometric, and polarimetric methods listed for various assays.

Urine Analysis

• Types of specimens: Random, first morning, postprandial, afternoon, day, night, and twenty-four-hours urine specimens.
• Physical properties: Volume, color, odor, reaction, specific gravity, and aspect.
• Chemical properties: Reducing sugars, ketones, proteins, urea, phosphates, calcium, bile salts, bile pigments, and urobilinogen.

DNA Extraction

• Definition: Removal of DNA from cells.
• Sources: Any living tissues.
• Method of Extraction: three steps; crushing the cells, extracting the buffer, and using ice-cold alcohol.
• Extraction Procedure: Steps involved in DNA isolation, including crushing, extracting buffer, cell filtration, alcohol addition, and removal of protein sediments to isolate pure DNA.

PCR

• Definition: A method of DNA cloning in a lab setting.
• Advantages: Simple, sensitive, powerful, and reliable technique, fast.
• Principle: Amplifies specific DNA sequences.
• Components: DNA template, DNA polymerase, primers, and dNTPs.
• Types of PCR: Conventional PCR, Multiplex PCR, Nested PCR, Reverse Transcriptase-PCR, Quantitative RT-PCR, and Hot start PCR; especially Multiplex RT-PCR for simultaneous analysis of different targets.
• Applications of PCR Medicine, including diagnosis of infectious diseases and genetic disorders; forensic examinations for paternity and criminal investigations; environmental studies, including detection of pathogens in water supplies; and agricultural applications, including genetic engineering.

Urine sheet

• Tests and results for various components in urine (volume, color, odor, reaction, specific gravity, aspect, reducing sugars, ketones using Rothera or Acetone test, Urinary Protein, Urea, Phosphate, calcium, bile salts, bile pigments, and urobilinogen)