Untitled Quiz
48 Questions
1 Views

Choose a study mode

Play Quiz
Study Flashcards
Spaced Repetition
Chat to Lesson

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

Which of the following is NOT a physiological cause of proteinuria?

  • Pregnancy
  • Severe muscular exercise
  • Prolonged exposure to cold
  • Nephritis (correct)

What is the primary product of protein catabolism?

  • Uric acid
  • Ammonia
  • Creatinine
  • Urea (correct)

Which test helps detect proteins in urine?

  • Refractometry
  • Biuret test
  • Heat coagulation test (correct)
  • Acid precipitation test

Which of the following actions is acceptable in the lab?

<p>Wearing gloves when handling blood (C)</p> Signup and view all the answers

Where is urea formed in the body?

<p>Liver (B)</p> Signup and view all the answers

What should you do if you notice broken glassware in the lab?

<p>Inform the instructor immediately (D)</p> Signup and view all the answers

What characterizes Bence Jones protein in urine?

<p>It clots when heated to 60°C (B)</p> Signup and view all the answers

Which non-protein nitrogenous compound is used in kidney function tests?

<p>Creatinine (A)</p> Signup and view all the answers

How should heated test tubes be handled?

<p>Point them away from yourself and others (D)</p> Signup and view all the answers

What is the normal plasma urea level?

<p>20-40 mg/dl (A)</p> Signup and view all the answers

In the event of a chemical spill, what is the immediate action to take?

<p>Clean all spills immediately (A)</p> Signup and view all the answers

What should you do if you experience a fainting episode in the lab?

<p>Provide fresh air and keep the head lower than the body (C)</p> Signup and view all the answers

Which of the following conditions is a pre-renal cause of proteinuria?

<p>Intraabdominal tumors (B)</p> Signup and view all the answers

When is it essential to wear safety goggles in the lab?

<p>When instructed by the lab supervisor (A)</p> Signup and view all the answers

What is the best practice regarding food and drinks in the lab?

<p>Food and drinks should be kept in bags (A)</p> Signup and view all the answers

What is the first thing you should do in case of an acid or alkali burn?

<p>Wash the affected area with plenty of running water (A)</p> Signup and view all the answers

What is the primary purpose of using detergent in the DNA extraction process?

<p>To dissolve the cell membrane (C)</p> Signup and view all the answers

Which component of the extraction buffer is specifically used to inhibit the nuclease enzyme?

<p>EDTA (C)</p> Signup and view all the answers

Why does the DNA precipitate when ice-cold alcohol is added to the solution?

<p>DNA is insoluble in alcohol. (A)</p> Signup and view all the answers

Which barrier is specifically present only in plant cells and must be overcome during DNA extraction?

<p>Cell wall (B)</p> Signup and view all the answers

What is the function of salt in the DNA extraction buffer?

<p>To stabilize the negatively charged phosphate groups of DNA (D)</p> Signup and view all the answers

In which step of DNA extraction is the tomato crushed to start the process?

<p>Step (1): Crushing (A)</p> Signup and view all the answers

Which of the following statements correctly describes what happens to the proteins during the DNA extraction process?

<p>They sink to the bottom of the test tube. (C)</p> Signup and view all the answers

What is the role of ice-cold alcohol in the DNA extraction protocol?

<p>To precipitate the DNA out of solution. (B)</p> Signup and view all the answers

What enzyme isoform is specifically elevated in myocardial infarction?

<p>CK2 (MB) (D)</p> Signup and view all the answers

What is the specific pattern of enzyme elevation in liver diseases?

<p>Very high ALT/AST with mild elevation of ALP (C)</p> Signup and view all the answers

How quickly does CK2 rise after the onset of pain in myocardial infarction?

<p>4-8 hours (D)</p> Signup and view all the answers

Which enzyme is more specific for liver damage when comparing ALT and AST?

<p>ALT (C)</p> Signup and view all the answers

Which isoenzyme is used for monitoring the follow-up of myocardial infarction?

<p>LDH1 (C)</p> Signup and view all the answers

What happens to CK2 levels after a myocardial infarction event?

<p>Returns to normal after 2-3 days (B)</p> Signup and view all the answers

Which liver condition is characterized by a rapid rise in transaminases before bilirubin elevation?

<p>Viral hepatitis (D)</p> Signup and view all the answers

In which muscle type is CK3 (MM) specifically located?

<p>Skeletal muscle (D)</p> Signup and view all the answers

What is a normal protein excretion ratio for children under 2 years old?

<p>&lt; 0.5 (D)</p> Signup and view all the answers

What does a protein ratio greater than 2 indicate?

<p>Nephrotic range proteinuria (D)</p> Signup and view all the answers

In electrophoresis, which of the following factors affects the migration of proteins?

<p>Charge of the molecules (C)</p> Signup and view all the answers

What is one of the clinical applications of electrophoresis?

<p>Plasma protein electrophoresis (B)</p> Signup and view all the answers

At what pH is serum protein electrophoresis performed?

<p>Alkaline pH (B)</p> Signup and view all the answers

Which of the following statements about plasma proteins in electrophoresis is correct?

<p>Proteins migrate towards the anode. (B)</p> Signup and view all the answers

What must be known to interpret the results of serum protein electrophoresis accurately?

<p>Common patterns of proteins (D)</p> Signup and view all the answers

Which of the following instruments is NOT typically used in electrophoresis?

<p>Spectrophotometer (B)</p> Signup and view all the answers

What is the primary purpose of primers in PCR?

<p>To initiate DNA synthesis on the target DNA (A)</p> Signup and view all the answers

Which of the following characteristics does Taq DNA polymerase possess?

<p>It has high processivity and does not exhibit proofreading (B)</p> Signup and view all the answers

In a standard PCR cycle, what occurs during the extension phase?

<p>Taq polymerase synthesizes new DNA strands (A)</p> Signup and view all the answers

What is the main advantage of using multiplex PCR?

<p>It allows for the simultaneous detection of multiple pathogens (B)</p> Signup and view all the answers

What is the purpose of reverse transcriptase in RT-PCR?

<p>To convert RNA into complementary DNA (C)</p> Signup and view all the answers

Which type of PCR specifically targets RNA for conversion to DNA?

<p>Reverse Transcriptase-PCR (A)</p> Signup and view all the answers

During which step of the PCR thermal cycle is the DNA separated into single strands?

<p>Denaturation (D)</p> Signup and view all the answers

What is one application of PCR in genetic engineering?

<p>To amplify genomic regions for sequencing (A)</p> Signup and view all the answers

Flashcards

Lab Behavior

Follow safety rules and procedures in the laboratory to prevent accidents and ensure a safe working environment.

Food/Drink in Lab

Eating and drinking are strictly prohibited in the lab. Food and drinks must be kept in bags.

Glassware Handling

Use clean, dry glassware. Check for cracks. Handle with holders when hot.

Chemical Handling

Never taste or touch chemicals directly. Never pour used reagents back into bottles.

Signup and view all the flashcards

Emergency Procedures (Burns)

Immediately flush affected area with plenty of water in case of acid or alkali burns.

Signup and view all the flashcards

Emergency Procedures (Eyes)

Irrigate eyes with plenty of water for at least 15 minutes in case of eye injuries.

Signup and view all the flashcards

Emergency Procedures (Fainting)

Provide fresh air and position the person with their head lower than their body in case of fainting.

Signup and view all the flashcards

Lab Cleanup

Thoroughly clean the laboratory surfaces before leaving and return equipment to its proper location

Signup and view all the flashcards

Protein excretion ratio in children under 2

Less than 0.5 in children less than 2 years old

Signup and view all the flashcards

Protein excretion ratio (children over 2)

Less than 0.2 in children over 2 years old

Signup and view all the flashcards

Nephrotic range proteinuria

Protein excretion ratio greater than 2

Signup and view all the flashcards

Electrophoresis

Migration of charged particles (like proteins) in an electric field

Signup and view all the flashcards

Plasma protein electrophoresis

Electrophoresis technique for separation of plasma proteins

Signup and view all the flashcards

Serum protein electrophoresis pH

Performed at an alkaline pH

Signup and view all the flashcards

Plasma protein migration

Negatively charged proteins migrate towards the anode

Signup and view all the flashcards

Normal plasma protein pattern interpretation

Interpretation of protein bands based on the normal distribution patterns for various proteins

Signup and view all the flashcards

Cell Wall Barrier

A rigid outer layer found in plant cells, protecting the cell and providing support.

Signup and view all the flashcards

Cell Membrane Barrier

The thin, flexible layer surrounding all cells, controlling what enters and exits.

Signup and view all the flashcards

Nuclear Membrane Barrier

The membrane surrounding the nucleus of a cell, regulating passage into and out.

Signup and view all the flashcards

Extraction Buffer

A solution containing detergent, salt, and EDTA used to break down cell membranes.

Signup and view all the flashcards

Detergent's Role

Dissolves the cell membranes (fatty component) to extract the DNA.

Signup and view all the flashcards

Salt's Role

Stabilizes DNA by neutralizing its negative charges to prevent its strands from repelling each other.

Signup and view all the flashcards

Ice Cold Alcohol's Role

Used to precipitate DNA making it visible and separate from other cellular components.

Signup and view all the flashcards

DNA Extraction Procedure

A series of steps—crushing, buffer treatment, and alcohol precipitation—to isolate DNA from a cell.

Signup and view all the flashcards

Proteinuria causes

Protein in the urine, caused by physiological (like prolonged standing) or pathological factors (pre-renal, renal, post-renal)

Signup and view all the flashcards

Pre-renal proteinuria

Protein in urine caused by conditions before the kidney, e.g., dehydration, heart failure.

Signup and view all the flashcards

Renal proteinuria

Protein in urine due to problems within the kidney, e.g., glomerulonephritis.

Signup and view all the flashcards

Post-renal proteinuria

Proteinuria caused by issues after the kidneys, e.g., urinary tract infections.

Signup and view all the flashcards

Bence Jones protein

Abnormal protein found in cancers such as multiple myeloma, affecting urine reactions.

Signup and view all the flashcards

Urea formation

Liver converts toxic ammonia into non-toxic urea, a crucial metabolic step.

Signup and view all the flashcards

Plasma Urea Level

Normal plasma urea levels range from 20-40 mg/dL, used to assess kidney function.

Signup and view all the flashcards

Heat coagulation test

A test for proteins in urine; if coagulation happens with heating, proteins are present (Albumin or Globulin).

Signup and view all the flashcards

Creatine Kinase (CK)

An enzyme that converts creatine and ATP into creatine phosphate and ADP, crucial for energy transfer in muscles.

Signup and view all the flashcards

CK Isoforms

Different forms of creatine kinase (CK), each specific to a type of muscle tissue (brain, heart, skeletal).

Signup and view all the flashcards

CK-MB in MI

CK-MB (heart-specific CK) significantly elevates in myocardial infarction (heart attack), used for early detection.

Signup and view all the flashcards

Myocardial Infarction (MI)

Occlusion of coronary arteries and heart muscle damage due to lack of blood flow.

Signup and view all the flashcards

ALT/AST in Liver Disease

Liver enzymes ALT and AST show elevated levels in liver damage, with ALT being more specific for liver injury.

Signup and view all the flashcards

Liver Enzyme Elevation in Viral Hepatitis

Viral hepatitis causes very rapid rises of liver enzymes (AST and ALT) in blood, before bilirubin elevation.

Signup and view all the flashcards

Time Course of CK-MB in MI

CK-MB levels rise within ~4-8 hours after MI symptoms, peak within 24 hours, and return to normal within 2-3 days.

Signup and view all the flashcards

LDH1 in MI Follow-up

LDH1 (Lactate Dehydrogenase) enzyme levels are used for long-term monitoring of MI, complementing CK-MB use.

Signup and view all the flashcards

PCR Primers

Short, synthetic DNA sequences that bind to specific target DNA sequences in a PCR reaction.

Signup and view all the flashcards

PCR Enzyme

Thermostable polymerase, usually Taq polymerase, that copies DNA during PCR, crucial for the amplification process.

Signup and view all the flashcards

PCR Denaturation

The step in PCR where heating separates the DNA double helix into two strands.

Signup and view all the flashcards

PCR Annealing

The step in PCR where the temperature is lowered so primers bind to their complementary target sequences.

Signup and view all the flashcards

PCR Extension

The step in PCR where DNA polymerase adds nucleotides to the primers, extending the DNA strands.

Signup and view all the flashcards

Multiplex PCR

A PCR technique used to detect multiple pathogens or DNA sequences in a single reaction.

Signup and view all the flashcards

Reverse Transcription PCR (RT-PCR)

A PCR technique that amplifies RNA sequences after converting them to cDNA.

Signup and view all the flashcards

PCR Applications

PCR has various uses, including genetic engineering (e.g., insertion of cloned DNA into organisms) and diagnostics (identifying pathogens, genetic mutations).

Signup and view all the flashcards

Study Notes

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)

Studying That Suits You

Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

Quiz Team

Related Documents

Practical Biochemistry 2024-2025 PDF

More Like This

Untitled Quiz
55 questions

Untitled Quiz

StatuesquePrimrose avatar
StatuesquePrimrose
Untitled Quiz
18 questions

Untitled Quiz

RighteousIguana avatar
RighteousIguana
Untitled Quiz
50 questions

Untitled Quiz

JoyousSulfur avatar
JoyousSulfur
Untitled Quiz
48 questions

Untitled Quiz

StraightforwardStatueOfLiberty avatar
StraightforwardStatueOfLiberty
Use Quizgecko on...
Browser
Open
Browser
Browser