INI-CET 2021 — Biochemistry

Q: Methyl-tetrahydrofolate (5-methyl-THF) gets accumulated in deficiency of which of the following?

Vitamin B12 (Cobalamin). ***Vitamin B12 (Cobalamin)*** - A deficiency in **Vitamin B12** (cobalamin) leads to the accumulation of **5-methyl-tetrahydrofolate (5-methyl-THF)** due to the **"folate trap"** hypothesis. - This occurs because B12 is a cofactor for **methionine synthase**, which converts 5-methyl-THF back to tetrahydrofolate (THF), a necessary step for DNA synthesis and other one-carbon metabolism reactions. - Without B12, folate remains trapped in the methyl form and cannot be utilized for other essential reactions. *Vitamin B2 (Riboflavin)* - **Vitamin B2** (riboflavin) is a precursor for **FAD** and **FMN**, essential coenzymes in various redox reactions in the electron transport chain and other metabolic pathways. - Its deficiency typically presents as **cheilosis**, glossitis, and angular stomatitis, but does not cause methyl-THF accumulation. *Vitamin B1 (Thiamine)* - **Vitamin B1** (thiamine) is a cofactor for enzymes like **pyruvate dehydrogenase** and **alpha-ketoglutarate dehydrogenase** in carbohydrate metabolism. - Its deficiency causes **Beriberi** (wet, dry, or Wernicke-Korsakoff syndrome), affecting the cardiovascular and nervous systems, but does not affect folate metabolism. *Vitamin B6 (Pyridoxine)* - **Vitamin B6** (pyridoxine) is a coenzyme for many metabolic reactions, particularly in amino acid metabolism and neurotransmitter synthesis. - Deficiency can lead to **sideroblastic anemia**, neurological symptoms, and dermatitis, but does not cause methyl-THF accumulation. [Practice more Biochemistry PYQs on OnCourse]

Q: Which of the following molecular analysis methods is used to detect RNA?

RT-PCR. ***RT-PCR*** - **Reverse transcriptase polymerase chain reaction (RT-PCR)** is a molecular method that first converts **RNA into complementary DNA (cDNA)** using reverse transcriptase before amplification via PCR. - This technique is commonly used to detect and quantify specific **RNA sequences**, such as viral RNA or gene expression levels. *Western blot* - **Western blot** is a laboratory technique used to detect specific **proteins** in a sample. - It involves separating proteins by size using gel electrophoresis, transferring them to a membrane, and then detecting the target protein using specific antibodies. *G banding* - **G banding** is a cytogenetic technique used to produce a visible **karyotype** by staining condensed chromosomes. - This method is primarily used to detect large-scale structural changes in chromosomes, not to detect RNA. *Sanger's method* - **Sanger's method**, also known as **dideoxy sequencing**, is a DNA sequencing technique. - It is used to determine the exact order of **nucleotides within a DNA molecule**, not to detect RNA directly. [Practice more Biochemistry PYQs on OnCourse]

Q: Glycated hemoglobin (HbA1c) is best measured using?

Ion exchange chromatography. ***Ion exchange chromatography*** - This method separates hemoglobin variants based on their **charge differences** due to the glucose molecule attached to HbA1c. - It is a highly sensitive and specific method for quantifying HbA1c, widely used in clinical laboratories. *Isoelectric focusing* - This technique separates molecules based on their **isoelectric point (pI)**, the pH at which they have no net charge. - While it can differentiate some hemoglobin variants, it is generally **less efficient and more complex** for routine HbA1c measurement compared to ion exchange chromatography. *Affinity chromatography* - This method separates molecules based on their **specific binding affinity** to a ligand immobilized on a stationary phase. - While it has been explored for HbA1c measurement, it is **not the most commonly used** or preferred method due to potential interferences and cost compared to ion exchange chromatography. *Electrophoresis* - This technique separates molecules based on their **charge and size** in an electric field. - While it can separate major hemoglobin variants, it has **lower resolution and accuracy** for routine HbA1c quantification compared to more specialized chromatographic methods, making it less ideal for precise measurement. [Practice more Biochemistry PYQs on OnCourse]

Q: Which of the following options is false in a patient with advanced liver disease?

Increased oxidative decarboxylation of alpha-ketoglutarate. ***Increased oxidative decarboxylation of alpha-ketoglutarate*** - In advanced liver disease, **hepatic encephalopathy** is a common complication due to the liver's inability to detoxify **ammonia**. - Alpha-ketoglutarate is crucial for detoxifying ammonia into glutamate and then glutamine, processes that involve reductive amination and **ATP consumption**, not increased oxidative decarboxylation. *Decreased ATP synthesis* - Advanced liver disease impairs various metabolic functions, including those of the **mitochondria**, leading to decreased ATP production. - This reduction in **energy reserves** affects numerous cellular processes and organ functions. *Hyperammonemia* - The liver is responsible for detoxifying **ammonia** into urea via the **urea cycle**. - In advanced liver disease, the impaired function of the liver leads to a buildup of ammonia in the blood, resulting in **hyperammonemia**. *Increased synthesis of glutamine and glutamate from alpha-ketoglutarate* - In the brain, when the liver fails to detoxify ammonia, **astrocytes** attempt to convert excess ammonia into **glutamine** using **alpha-ketoglutarate**. - This process is a compensatory mechanism to reduce ammonia toxicity, but it depletes alpha-ketoglutarate, which is a key intermediate in the **Krebs cycle**, leading to **cerebral energy deficits**. [Practice more Biochemistry PYQs on OnCourse]

Q: Which of the following is not a substrate for glucose formation?

Acetyl coenzyme A. ***Acetyl coenzyme A*** - **Acetyl CoA** cannot be converted to glucose because the two carbons from the acetyl group are lost as carbon dioxide in the **Krebs cycle**, making a net synthesis of glucose impossible. - The irreversible nature of the **pyruvate dehydrogenase complex** prevents the conversion of Acetyl CoA back to **pyruvate**, which is a crucial step for gluconeogenesis. *Lactate* - **Lactate** is a major substrate for gluconeogenesis, particularly during exercise and fasting, via the **Cori cycle**. - **Lactate dehydrogenase** converts lactate to **pyruvate**, which can then enter the gluconeogenic pathway. *Glycerol* - **Glycerol**, derived from triglyceride breakdown, enters gluconeogenesis by being converted to **glycerol-3-phosphate** and then to **dihydroxyacetone phosphate (DHAP)**. - DHAP is an intermediate in glycolysis and gluconeogenesis, allowing for its conversion to glucose. *Alanine* - **Alanine** is a **glucogenic amino acid** that can be transaminated to **pyruvate**. - **Pyruvate** can then proceed through the gluconeogenic pathway to synthesize glucose, especially during prolonged fasting. [Practice more Biochemistry PYQs on OnCourse]

11 Previous Year Questions with Answers & Explanations

Questions

Methyl-tetrahydrofolate (5-methyl-THF) gets accumulated in deficiency of which of the following?

Which of the following molecular analysis methods is used to detect RNA?

Glycated hemoglobin (HbA1c) is best measured using?

Which of the following options is false in a patient with advanced liver disease?

Which of the following is not a substrate for glucose formation?

What is the repair mechanism associated with CRISPR-cas9?

Which of the following is not amphipathic?

Beta hCG is structurally similar to which biochemical moiety?

miRNA binds to which part of the mRNA to inhibit translation?

Q10

Which of the following methods cannot be used to precipitate proteins?

INI-CET 2021 - Biochemistry INI-CET Practice Questions and MCQs

Question 1: Methyl-tetrahydrofolate (5-methyl-THF) gets accumulated in deficiency of which of the following?

A. Vitamin B12 (Cobalamin) (Correct Answer)
B. Vitamin B2 (Riboflavin)
C. Vitamin B1 (Thiamine)
D. Vitamin B6 (Pyridoxine)

Explanation: ***Vitamin B12 (Cobalamin)*** - A deficiency in **Vitamin B12** (cobalamin) leads to the accumulation of **5-methyl-tetrahydrofolate (5-methyl-THF)** due to the **"folate trap"** hypothesis. - This occurs because B12 is a cofactor for **methionine synthase**, which converts 5-methyl-THF back to tetrahydrofolate (THF), a necessary step for DNA synthesis and other one-carbon metabolism reactions. - Without B12, folate remains trapped in the methyl form and cannot be utilized for other essential reactions. *Vitamin B2 (Riboflavin)* - **Vitamin B2** (riboflavin) is a precursor for **FAD** and **FMN**, essential coenzymes in various redox reactions in the electron transport chain and other metabolic pathways. - Its deficiency typically presents as **cheilosis**, glossitis, and angular stomatitis, but does not cause methyl-THF accumulation. *Vitamin B1 (Thiamine)* - **Vitamin B1** (thiamine) is a cofactor for enzymes like **pyruvate dehydrogenase** and **alpha-ketoglutarate dehydrogenase** in carbohydrate metabolism. - Its deficiency causes **Beriberi** (wet, dry, or Wernicke-Korsakoff syndrome), affecting the cardiovascular and nervous systems, but does not affect folate metabolism. *Vitamin B6 (Pyridoxine)* - **Vitamin B6** (pyridoxine) is a coenzyme for many metabolic reactions, particularly in amino acid metabolism and neurotransmitter synthesis. - Deficiency can lead to **sideroblastic anemia**, neurological symptoms, and dermatitis, but does not cause methyl-THF accumulation.

Question 2: Which of the following molecular analysis methods is used to detect RNA?

A. Western blot
B. G banding
C. RT-PCR (Correct Answer)
D. Sanger's method

Explanation: ***RT-PCR*** - **Reverse transcriptase polymerase chain reaction (RT-PCR)** is a molecular method that first converts **RNA into complementary DNA (cDNA)** using reverse transcriptase before amplification via PCR. - This technique is commonly used to detect and quantify specific **RNA sequences**, such as viral RNA or gene expression levels. *Western blot* - **Western blot** is a laboratory technique used to detect specific **proteins** in a sample. - It involves separating proteins by size using gel electrophoresis, transferring them to a membrane, and then detecting the target protein using specific antibodies. *G banding* - **G banding** is a cytogenetic technique used to produce a visible **karyotype** by staining condensed chromosomes. - This method is primarily used to detect large-scale structural changes in chromosomes, not to detect RNA. *Sanger's method* - **Sanger's method**, also known as **dideoxy sequencing**, is a DNA sequencing technique. - It is used to determine the exact order of **nucleotides within a DNA molecule**, not to detect RNA directly.

Question 3: Glycated hemoglobin (HbA1c) is best measured using?

A. Isoelectric focusing
B. Affinity chromatography
C. Electrophoresis
D. Ion exchange chromatography (Correct Answer)

Explanation: ***Ion exchange chromatography*** - This method separates hemoglobin variants based on their **charge differences** due to the glucose molecule attached to HbA1c. - It is a highly sensitive and specific method for quantifying HbA1c, widely used in clinical laboratories. *Isoelectric focusing* - This technique separates molecules based on their **isoelectric point (pI)**, the pH at which they have no net charge. - While it can differentiate some hemoglobin variants, it is generally **less efficient and more complex** for routine HbA1c measurement compared to ion exchange chromatography. *Affinity chromatography* - This method separates molecules based on their **specific binding affinity** to a ligand immobilized on a stationary phase. - While it has been explored for HbA1c measurement, it is **not the most commonly used** or preferred method due to potential interferences and cost compared to ion exchange chromatography. *Electrophoresis* - This technique separates molecules based on their **charge and size** in an electric field. - While it can separate major hemoglobin variants, it has **lower resolution and accuracy** for routine HbA1c quantification compared to more specialized chromatographic methods, making it less ideal for precise measurement.

Question 4: Which of the following options is false in a patient with advanced liver disease?

A. Decreased ATP synthesis
B. Hyperammonemia
C. Increased synthesis of glutamine and glutamate from alpha-ketoglutarate
D. Increased oxidative decarboxylation of alpha-ketoglutarate (Correct Answer)

Explanation: ***Increased oxidative decarboxylation of alpha-ketoglutarate*** - In advanced liver disease, **hepatic encephalopathy** is a common complication due to the liver's inability to detoxify **ammonia**. - Alpha-ketoglutarate is crucial for detoxifying ammonia into glutamate and then glutamine, processes that involve reductive amination and **ATP consumption**, not increased oxidative decarboxylation. *Decreased ATP synthesis* - Advanced liver disease impairs various metabolic functions, including those of the **mitochondria**, leading to decreased ATP production. - This reduction in **energy reserves** affects numerous cellular processes and organ functions. *Hyperammonemia* - The liver is responsible for detoxifying **ammonia** into urea via the **urea cycle**. - In advanced liver disease, the impaired function of the liver leads to a buildup of ammonia in the blood, resulting in **hyperammonemia**. *Increased synthesis of glutamine and glutamate from alpha-ketoglutarate* - In the brain, when the liver fails to detoxify ammonia, **astrocytes** attempt to convert excess ammonia into **glutamine** using **alpha-ketoglutarate**. - This process is a compensatory mechanism to reduce ammonia toxicity, but it depletes alpha-ketoglutarate, which is a key intermediate in the **Krebs cycle**, leading to **cerebral energy deficits**.

Question 5: Which of the following is not a substrate for glucose formation?

A. Lactate
B. Glycerol
C. Alanine
D. Acetyl coenzyme A (Correct Answer)

Explanation: ***Acetyl coenzyme A*** - **Acetyl CoA** cannot be converted to glucose because the two carbons from the acetyl group are lost as carbon dioxide in the **Krebs cycle**, making a net synthesis of glucose impossible. - The irreversible nature of the **pyruvate dehydrogenase complex** prevents the conversion of Acetyl CoA back to **pyruvate**, which is a crucial step for gluconeogenesis. *Lactate* - **Lactate** is a major substrate for gluconeogenesis, particularly during exercise and fasting, via the **Cori cycle**. - **Lactate dehydrogenase** converts lactate to **pyruvate**, which can then enter the gluconeogenic pathway. *Glycerol* - **Glycerol**, derived from triglyceride breakdown, enters gluconeogenesis by being converted to **glycerol-3-phosphate** and then to **dihydroxyacetone phosphate (DHAP)**. - DHAP is an intermediate in glycolysis and gluconeogenesis, allowing for its conversion to glucose. *Alanine* - **Alanine** is a **glucogenic amino acid** that can be transaminated to **pyruvate**. - **Pyruvate** can then proceed through the gluconeogenic pathway to synthesize glucose, especially during prolonged fasting.

Question 6: What is the repair mechanism associated with CRISPR-cas9?

A. Mismatch repair
B. Non-homologous end joining (Correct Answer)
C. Nucleotide excision repair
D. Base excision repair

Explanation: ***Non-homologous end joining*** - **CRISPR-Cas9** creates a **double-strand break (DSB)** in DNA, which is primarily repaired by **non-homologous end joining (NHEJ)**. - **NHEJ** is the **predominant repair pathway** in most cells, accounting for 60-90% of DSB repairs. - **NHEJ** is an error-prone repair mechanism that ligates the broken ends directly, often leading to small **insertions or deletions (indels)** causing gene knockout. - Note: **Homology-directed repair (HDR)** is another CRISPR-associated mechanism used for precise editing when a donor template is provided, but **NHEJ is the primary endogenous repair pathway**. *Mismatch repair* - This mechanism corrects errors that arise during **DNA replication**, such as incorrect base pairing. - It does not repair **double-strand breaks** induced by CRISPR-Cas9. *Nucleotide excision repair* - This pathway removes **bulky DNA adducts** and lesions, such as those caused by UV radiation. - It is not involved in repairing **CRISPR-Cas9 induced double-strand breaks**. *Base excision repair* - This mechanism corrects **damaged or modified bases**, typically single base changes, without affecting the sugar-phosphate backbone significantly. - It handles different types of DNA damage than the **double-strand breaks** generated by CRISPR-Cas9.

Question 7: Which of the following is not amphipathic?

A. Triglycerides (Correct Answer)
B. Sphingolipids
C. Glycolipids
D. Phosphoglycerol

Explanation: ***Triglycerides*** - Triglycerides are composed of a **glycerol backbone** esterified to three fatty acids, making them entirely **hydrophobic** and thus not amphipathic. - They serve primarily as **energy storage** molecules and do not form membranes because they lack a polar head group. *Sphingolipids* - Sphingolipids are amphipathic because they contain a **hydrophilic polar head group** (e.g., phosphocholine or a sugar) and two **hydrophobic tails** derived from a fatty acid and the sphingosine backbone. - This dual nature allows them to be fundamental components of **cell membranes**. *Glycolipids* - Glycolipids are characterized by a **carbohydrate head group** attached to a lipid moiety, rendering them amphipathic. - The sugar portion is **hydrophilic**, while the lipid portion (e.g., ceramide) is **hydrophobic**, enabling them to participate in cell recognition and membrane stability. *Phosphoglycerol* - Phosphoglycerol (more commonly referred to as **glycerophospholipids**) are amphipathic, consisting of a **glycerol backbone**, two fatty acid tails, and a **phosphate group** often linked to an alcohol. - The **phosphate and alcohol group** form the hydrophilic head, and the **fatty acid tails** form the hydrophobic region, making them crucial for lipid bilayers.

Question 8: Beta hCG is structurally similar to which biochemical moiety?

A. LH (Correct Answer)
B. Oxytocin
C. FSH
D. ACTH

Explanation: ***LH*** - **Beta-hCG** and **Luteinizing Hormone (LH)** both belong to the family of glycoprotein hormones and share a common alpha subunit. - Their **beta subunits** are also very similar, with a significant overlap in their amino acid sequences, allowing **hCG** to bind to and activate **LH receptors**. *Oxytocin* - **Oxytocin** is a **peptide hormone** primarily involved in uterine contractions and milk ejection. - It is structurally distinct from **glycoprotein hormones** like **hCG** and lacks the alpha and beta subunit structure. *FSH* - **Follicle-Stimulating Hormone (FSH)** is a glycoprotein hormone that shares the same alpha subunit with hCG, LH, and TSH. - However, its **beta subunit** is distinct from that of **hCG**, which gives it its unique biological specificity. *ACTH* - **Adrenocorticotropic Hormone (ACTH)** is a **peptide hormone** produced by the anterior pituitary gland, regulating adrenal gland function. - It has a completely different structure and function compared to **hCG** and other glycoprotein hormones.

Question 9: miRNA binds to which part of the mRNA to inhibit translation?

A. Gene promoter
B. 3'UTR (Correct Answer)
C. Gene body
D. 5'UTR

Explanation: ***3'UTR*** - MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene expression. - They primarily bind to the **3' untranslated region (3'UTR)** of messenger RNA (mRNA) molecules, leading to translational repression or mRNA degradation. *Gene promoter* - The **gene promoter** is a region of DNA located upstream of a gene, where regulatory proteins bind to initiate transcription. - miRNAs do not directly bind to gene promoters to inhibit translation. *Gene body* - The **gene body** refers to the entire transcribed region of a gene, including exons and introns. - While some regulatory elements can be found within the gene body, the primary binding site for miRNAs to exert translational control is the 3'UTR. *5'UTR* - The **5' untranslated region (5'UTR)** is located at the 5' end of an mRNA molecule, upstream of the start codon. - While the 5'UTR can play a role in regulating translation initiation, it is not the primary target for miRNA binding to inhibit translation.

Question 10: Which of the following methods cannot be used to precipitate proteins?

A. Add alcohol and acetone
B. Using heavy metal ions
C. Adding trichloroacetic acid
D. Moving pH away from isoelectric pH (Correct Answer)

Explanation: ***Moving pH away from isoelectric pH*** - Proteins are **least soluble** at their **isoelectric point (pI)**, where their net charge is zero, causing them to aggregate and precipitate. - Moving the pH **away from the isoelectric point** increases the net charge on the protein, enhancing its solubility and preventing precipitation. *Add alcohol and acetone* - **Organic solvents** like alcohol and acetone reduce the dielectric constant of water, weakening the **hydrophobic interactions** that maintain protein solubility. - This leads to increased protein-protein interactions and **precipitation** as the protein unfolds or aggregates. *Using heavy metal ions* - **Heavy metal ions** (e.g., lead, mercury) are positively charged and bind strongly to the negatively charged groups on proteins, such as **carboxylates** and **sulfhydryl groups**. - This binding can disrupt protein structure, lead to aggregation, and cause **precipitation**. *Adding trichloroacetic acid* - **Trichloroacetic acid (TCA)** is a strong acid that significantly lowers the pH of the solution, causing proteins to become **protonated**. - This change in charge and the disruption of **salt bridges** and hydrogen bonds lead to protein denaturation and **precipitation**.