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Ehrlichs reagent is for urobilinogen as well as porphobilinogen !!! Ehrlichs diazo reagent: Two solutions, one of sodium nitrite, the other of acidified sulphanilic acid, used in bringing about diazotisation. This is reagent used in the Van de Bergh reaction used to estimate bilirubin (Direct and Indirect) in blood. Ehrlichia: Small, often pleomorphic, coccoid to ellipsoidal organisms occurring intracytoplasmically in circulating lymphocytes. They are the aetiologic agents of tick-borne diseases of humans, dogs, cattle, sheep, goats, and horses. Ehrlichs anaemia: Aplastic anaemia Ehrlichs inner body: Heinz-Ehrlich body : A round oxyphil body found in the red blood cell in case of haemocytolysis due to a specific blood poison. Ehrlichs phenomenon: The difference between the amount of diphtheria toxin that will exactly neutralise one unit of antitoxin and that which, added to one unit of antitoxin, will leave one lethal dose free is greater than one lethal dose of toxin; i.e., it is necessary to add more than one lethal dose of toxin to a neutral mixture of toxin and antitoxin to make the mixture lethal (the basis of the L+ dose). Ehrlichs postulate: Side-chain theory : Ehrlich postulated that cells contained surface extensions or side chains (haptophores) that bind to the antigenic determinants of a toxin (toxophores); after a cell is stimulated, the haptophores are released into the circulation and become the antibodies. Ehrlich-turk line: Seldom-used term for the vertical, thin deposition of material on the posterior surface of the cornea in uveitis. Ehrlichs triacid stain: A differential leukocytic stain comprised of saturated solutions of orange G, acid fuchsin, and methyl green. 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Propoxur Ref::Trip 5th Page 81 Park 550 table Quality:reader Discussion Explanation: anticholinesterases Reversible Carbamates Acridine Physostigmine (Eserine) Neostigmine Pyridostigmine Edrophonium Rivastigmine Donepezil tacrine Irreversible Organophosphates Carbamates Dyflos DFP Echothiophate Parathion Malathion Diazinon TIK 20 Tabun Sarin Soman Carbaryl (Sevin) Propoxur(Baygon) Explanation A. dieldrin-organochlorine pesticide. incorrect choice. B. parathionirreversible OPC. correct choice. C. malathion irreversible OPC. correct choice D. Kepone/chlordecone- organochlorine pesticide. incorrect choice pesticide 5. Propoxurirreversible carbamates. incorrect choice Correct choices: b c 203. Which of the following are features of spinal shock a. Spasticity b. Urinary retention c. Areflexia d. Sensory level e. Increased DTRs Ref::clinical acumen Quality:spotter thinker reader BATA Discussion Spinal shock(neuronal) is the initial state just after an acute insult causing an UMN lesion caused by the temporary depressant effect on the anterior horn cells. Characterized by paralysis,areflexia,flaccidity,the chracteristic hypertonia and increased relexes of cotticospinal lesion appear after few hrs-days. Explanation Correct choices B C d 217. Which of the following predispose a patient to prehepatic encepalopathy a. Constipation b. Bacterial peritonitis c. Hemorrhage d. Hyperkalemia e. Dehydration Ref: HPIM 1715 Quality:spotter thinker reader BATA Discussion Common precipitants of hepatic failure Increased nitrogen load GIT bleeding Excess dietary protein Azotemia Constipation Electrolyte & metabolic imbalance Hypokalemia Alkalosis Hypoxia Hyponatremia Drugs Narcotics and Tranqulizers Sedatives Diuretics Miscellaneous Infection surgery Superimposed acute liver disease Progressive liver disease Explanation: A. Constipation B. Bacterial peritonitis being an infection. correct choice. C. Hemorrhage D. Hyperkalemia: Hypokalemia not Hyperkalemia is known to worsen hepatic Encephalopathy. 5. Dehydration can cause hypovolemia & electrolyte disturbances that can ppt. hepatic Encephalopathy Correct choices: a b c e 8. the drugs useful in the treatment of ectopic pregnancy is A. Methotrexate B. Actinomycin D C. Cyclophosphamide D. Potassium fluoride 5. Adriamycin Ref:: Dutta 202 Quality: spotter Explanation A. Methotrexate: used in medical therapy. correct choice. B. Actinomycin D. correct choice. C. Cyclophosphamide. Incorrect choice. D. Potassium fluoride: potassium chloride is used in ectopic pregnancy. Incorrect choice. 5. Adriamycin is not used in ectopic pregnancy. Correct choices A b 210. Adventitial bursitis due to tuberculosis is found in which of the following sites commonly a. Prepatellar b. Greater trochanter of femur c. Sub acromial d. Metatarsal e. Sub olecranon 209. Snow ball opacities in vitreous is seen in which of the following a. Parsplanitis b. Rheumatoid arthritis c. Anterior unietis d. Retinitis pigmentosa e. Endothelial dystrophy Ref: http://www. indmedica. com/cos/journal/vasculitis. html Quality:spotter thinker reader BATA Explanation A. Parsplanitis is characterized by snowball opacities. B. Rheumatoid arthritis is one of the causes of pars planitis hence may show snow ball opacities C. Anterior uveitis D. Retinitis pigmentosa 5. Endothelial dystrophy Correct choices: A b 77. The left border of the heart is formed by a. Pulmonary Artery b. Pulmonary vein c. Right ventricle d. Ascending aorta e. Arch of aorta Ref:: BDC 226,Grays 38th 1476 Quality: spotter thinker reader BATA Discussion Anatomically Right border is formed by RA. Left border of heart is formed by LA and LV. Upper border formed by LA, asc aota & pul trunk infron of it. Inferior border of the heart is formed by RV and contribution of LV at the apex. Correct choices NONE OF THE CHOICES ARE CORRECT Or 77. left border of the heart in chest XR is formed by a. pul artery b. pul vein c. abdominal aorta d. arch of aorta e. right ventricle Quality: :reader Reference: Discussion: Borders of cardiac silhouette on PA view Right border from above downwards Superior vena cava Outer border of R atrium Sometimes asc, arch of aorta & inferior vena cava Left border from above downwards Aortic knuckle Pul. Trunk or L pul artery L atrial appendage outer border of L ventricle correct choice: a d 9. the resting membrane potential depends on which of the following ions A. Magnesium B. Calcium C. Potassium D. Sodium 5. Chloride Ref:: Ganong 50 Quality: spotter Discussion The ionic basis of RMP: Explanation A. Magnesium has no role in RMP. B. Calcium has role in action potential in muscle cells not RMP C. Potassium. The major ion whose movements produce RMP. Correct choice. D. Sodium. incorrect choice. 5. Chloride. No role. Correct choices C 79. In acute inflammation the migration of neutrophils and its attachment to endothelium is mediated by which of the follwing. a. Integrins b. Selection s c. Adhesins d. Perforins e. Opsonins Ref:: Robbins 29, 273 Quality:spotter thinker reader BATA Explanation a. Integrins are transmembrane glycoproteins that function as cell receptors for extra cellular matrix. correct choice. b. Selectins=the loose and transient adhesions involved in rolling of leucocytes over endothelial cells are accounted for by the selectin family of molecules. E-selectin are present on on endothelial cells, P-selectin on platelets and endothelial cells and L-Selectins on leucocytes. c. Adhesins-are molecules that bind bacteria to cells e. g. lipoteichoic acid and protein F. d. Two classes of cytotoxins include perforins and granzymes. . The perforins form pores in the targets cell membranes. incorrect choice. e. Opsonins are protiens or peptides that label targets for phagocytosis by PMNs and/or macrophages. incorrect choice, Correct choices b 102. Susruta samhita was translated by a. Celsius b. harnel c. Bhargabhatta d. heslar e. Bernard Ref: http://www. ayurveda. com/online%20resource/ancient_writings. htm Quality: BATA Discussion: http://www. ayurveda. com/online%20resource/ancient_writings. htm Sushruta Samhita The Sushruta Samhita deals with the practice and theory of surgery, is an important source of Ayurvedic aphorisms. This work is the first to enumerate and discuss the pitta sub-doshas and the marmas. With its emphasis on pitta, surgery, and blood, this work best represents the transformational value of life. It is dais that this work was first redacted by Nagarjuna. This work, originally written in Sanskrit, is now available in English with Devanagari. Bhishagratnas translation is English and Sanskrit. P. V. Sharma has recently written a translation with both the Sanskrit/Devanagari and English that includes Dallanas commentary. Dallana has been regarded as the most influential commentator on Sushrutas work. Explanation Correct choices B d 21. Prozone phenomenon in precipitation reaction indicates a. False positive reaction b. False negative reaction c. due to excess antibodies d. Due to excess antigens e. Broken antibodies Ref: : AN 91 Quality: spotter 203. Which of the following are features of spinal shock f. Spasticity g. Urinary retention h. Areflexia i. Sensory level j. Increased DTRs Ref::clinical acumen Quality:spotter thinker reader BATA Discussion Spinal shock(neuronal) is the initial state just after an acute insult causing an UMN lesion caused by the temporary depressant effect on the anterior horn cells. Characterized by paralysis,areflexia,flaccidity,the chracteristic hypertonia and increased relexes of cotticospinal lesion appear after few hrs-days. Explanation 6. Spasticity appears hrs day after spinal shock has passed in an UMN lesion. 7. Urinary retention is a feature of spinal cord disease. appears first in case of disease originating from the canal and spreading outwards and last in mass lesions compressing spinal cord from the outside. Its a feature of spinal shock. 8. Areflexia is a feature of spinal shock. 9. Sensory level is seen in case of corticospinal lesion. 10. Increased DTRs will be seen after the spinal shock has passed. Correct choices B C d 202. Diseases inherited with triple repeat sequences include a. Huntingtons chorea b. Alzheimers disease c. Spinocerebellar ataxia d. Amyotrophic lateral sclerosis e. Ataxia telangiectasia Ref:: Robbins 197,739,741,157 Quality:spotter thinker reader BATA Discussion Harrison 2294 table gives explicit information about genetic defects in neurological disease. Robbins has enumerated three diseases having triple repeat sequences: Fragile X syndrome Huntington;s ds, Myotonic dystrophy Explanation A. Huntingtons chorea:AD : CAG triple repeats in gene encoding proein huntingtin B. Alzheimers disease:AD mutations presenilin,apolipoprotein E4,APP genen C. Spinocerebellar ataxia:expansion of trinucleotide repeats in ataxin gene D. Amyotrophic lateral sclerosis:AD mutation in Cu-Zn superoxide Dismutase SOD. 5. Ataxia telangiectasia AR mutation in ATM gene which causes triple repeat sequences and abnormal production of protein. Correct choices A c e 59. Tumor markers of hepatocellular carcinoma include a. CEA b. Des gamma carboxyprothrombin c. CA 19-9 d. Fucosylated alpha protein e. Alpha fetoprotien elevated in greater than 70% Ref:: HPIM 579, CMDT 689 http://www. intouchlive. com/consults/ccn9903b. htm Quality:spotter thinker reader BATA Discussion Alpha fetoprotein levels raised in Cirrhosis Massive liver necrosis Chronic hepatitis hepatocellular carcinoma Normal pregnancy Fetal distress or death Fetal neural tube defects-enencephaly & spina bifida Gonadal germ cell tumours teratoblastoma, Levels above 1000 ng/ml HCC CEA (Carcino Embryonic Antigen) elevated in: malignancies of the gastrointestinal tract (pancreas, colon, rectum), lung, breast, prostatic and ovary. inflammation and heavy smoking. CA-125 normally found in adult female fallopian tube, endometrium and endocervix. CA-15 elevated breast and lung cancer. . CA-199 elevated in intra-abdominal carcinomas, adenocarcinomas of the lung, gastric, biliary and colonic neoplasms. Explanation a. serum CEA levels alone are not specific for hepatocellular carcinoma but are nonetheless raised. correct choice. b. Des gamma carboxyprothrombin, an abnormal typre of [rothrombin which correlates with h levels of AFP. c. CA 19-9 levels are elevated in HCC. D most of the patients with hepatocellular carcinoma have an elevated serum concentration of fucosylated alpha-fetoprotein. correct choice. e. Alpha-fetoprotein elevated in greater than 70% . true. Correct choices A bcde 67. Which of the following are causes of posterior mediastinal mass: A. Neuroenteric cyst B. Neurogenic cyst C. Anterior thoraxic meningocele D. Lymphoma E. Bronchogenic cyst Anterior mediastinum: Middle Mediastinum: Posterior mediastinum: Germ cell neoplasm Lymphoma: Morgagni hernia Pericardial cyst Thymic lesions: Thyroid (Retrosternal goiter) Foregut duplication cysts Castlemans disease Lymphangioma Lymphoma Bronchogenic cyst Bochdalek Hernia Extramedullary Hematopoiesis Ganglioneuroma Ganglioneuroblastoma Neuroblastoma Lateral thoracic meningocele Malignant tumor of nerve sheath origin Neurofibroma Paraganglioma Pheochromocytoma Schwannoma Explanation A. Neuroenteric cyst is present in the posterior mediastinum or paravertebral area. B. Neurogenic cyst is present in the posterior mediastinum C. Anterior thoracic meningocele is present in the posterior mediastinum D. Lymphoma is a tumour of the middle or anterior mediastinum E. Bronchogenic cyst is a tumour of the middle mediastinum. Correct choices Abc 213. Peripheral neuropathy is seen in all of the following except a. Tuberculosis b. Polyarteritis nodasa c. Diabetes mellitus d. SLE 89. Diabetes mellitus is associated with all of the following except: a. Neuropathy b. Encephalopathy c. Myopathy d. Myelitis e. Myelopathy Ref: CMDT 1191,HPIM 2481 Quality:spotter thinker reader BATA Discussion Diabetic neuromuscular involvement may manifest as: A. Distal symmetric polyneuropathy B. Isolated peripheral neuropathy-mononeuropathy C. Mononeuritis multiplex D. Painful neuropathy 5. Autonomic neuropathy 6. Cranial neuropathypainful opthalmoplegia 7. Diabetic amyotrophy/proximal neuropathy/lumbosacral plexopathy 8. Ischemic myopathy of thigh muscles D e 127. In carbon monoxide toxicity a. There is left shift of oxygen dissociation curve b. Saturation of oxygen at 50% PaO2 is decreased c. Oxygen content of arterial blood is decreased d. Induces a state of metabolic acidosis e. Oxygen is used in treatment Ref: Ganong 17th 633 Quality:spotter thinker reader BATA Discussion Explanation a. There is left shift of oxygen dissociation curve. COHb shifts the curve of the remaining HbO2 to left decreasing the amount of O2 released by the normal saturated Hb. b. Saturation of oxygen at 50% PaO2 is decreased. I think this choice should be saturation of Hb at given PO2 is decreased which is untrue because the curve shifts to the left. This increases the saturation of Hb at givenPO2 and even makes the available HbO2 useless for oxygenation because it wont release the O2. c. Carboxu Hb reduces oxygen transportation by Hb thus Oxygen content of arterial blood is decreased. correct. d. Induces a state of metabolic acidosis because of the hypoxia e. Ventilation with O2 is preferable to ventilation with fresh air since O2 hastens the dissociation of COHb. Hyperbaric O2 is useful. 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Answer: D (X-ray diffraction) Ref: Harper, 25th edn, 56 may appear to indicate NMR; Harper 25th edn, 55; Harper 24th edn, 10 : The most detailed information is by X-Ray Diffraction and Crystallography; Harrison 15th edn,Chapter 322 -Gout and other crystal arthropathies; Harrison 15th edn,Chapter 327 Principles of Endocrinology; Campbells Urology 8th edn,3274; Mention option D as the investigating modality in revealing structures of microcrystal Quality: SpotterIf you had already read the references shown above Status: New QTDF: Harrison Discussion The use of polarising microscopy during synovial fluid analysis and the application of other crystallographic techniques, such as electron microscopy, energy-dispersive elemental analysis, and X-ray diffraction, have established the role of different microcrystals, including monosodium urate (MSU), calcium pyrophosphate dihydrate (CPPD), calcium hydroxyapatite (HA), and calcium oxalate (CaOx), in inducing acute or chronic arthritis or periarthritis. The family of nuclear receptors has grown to nearly 100 members, many of which are still classified as orphan receptors because their ligands, if they exist, remain to be identified. Otherwise, most nuclear receptors are classified based on the nature of their ligands. Though all nuclear receptors ultimately act to increase or decrease gene transcription, some (e.g., glucocorticoid receptor) reside primarily in the cytoplasm, whereas others (e.g., thyroid hormone receptor) are always located in the nucleus. After ligand binding, the cytoplasmically localised receptors translocate to the nucleus. The structures of nuclear receptors have been extensively studied, including by X-ray crystallography. X-ray crystallography shows that various SERMs induce distinct receptor conformations. Explanation A. Fluorescence spectroscopy: When a primary X-ray excitation source from an X-ray tube or a radioactive source strikes a sample, the X-ray can either be absorbed by the atom or scattered through the material. The process in which an X-ray is absorbed by the atom by transferring all of its energy to an innermost electron is called the photoelectric effect. During this process, if the primary X-ray had sufficient energy, electrons are ejected from the inner shells, creating vacancies. These vacancies present an unstable condition for the atom. As the atom returns to its stable condition, electrons from the outer shells are transferred to the inner shells and in the process give off a characteristic X-ray whose energy is the difference between the two binding energies of the corresponding shells. Because each element has a unique set of energy levels, each element produces X-rays at a unique set of energies, allowing one to non-destructively measure the elemental composition of a sample. The process of emissions of characteristic X-rays is called X-ray Fluorescence, or XRF. Analysis using X-ray fluorescence is called X-ray Fluorescence Spectroscopy. In most cases the innermost K and L shells are involved in XRF detection. A typical X-ray spectrum from an irradiated sample will display multiple peaks of different intensities. This is similar to the absorption spectrometry we use for determining the various forms of haemoglobin (given in Chaterjee Biochemistry and Narayana Reddy FM Blood stains) but instead of the visible light we use in Hb determination, we use X-rays here B. Electron microscopy: Electron microscopes are scientific instruments that use a beam of highly energetic electrons to examine objects on a very fine scale. 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NADPH-cytochrome P-450 reductase is involved in the reactivation of cytochrome P-450 which is not a stranger. B. Glutathione S-transferase is an enzyme involved in activation of xenobiotics, Harper has clearly stated that of all Phase 1 reactions it is hydroxylation (mono-oxygenation - Monooxygenatiuon means adding one oxygen. When that oxygen is added to a R-H group, we get R-OH and because of that -OH we call it as hydroxylation) which is important. Hence, there is no doubt regarding this enzyme is not the main enzyme. C. NADPH cytochrome P-450-reductase.is involved, but not in activation of xenobiotics: Xenobiotics is (Gk xenos stranger) is a compound that is foreign to the body, the principal classes of xenobiotics of medical relevance are drugs, chemical carcinogens. Obviously, P-450 is not a xenobiotic !!!! Then how come an enzyme (which activates another enzyme, which is in our body) be a main enzyme in the activation of xenobiotics !!!! Again the question is clear.. Activation of xenobiotics. Atleast if the question is concerned with xenobiotics,.. there can be a little confusion. D. Glucuronyl transferase is an enzyme involved in activation of xenobiotics, but it is not the main enzyme. Comments The confusa in this question is due to the line NADPH, not NADH is involved in the reaction mechanism of cytochrome P-450 in Harper. But as explained, NADPH and NADH is used, but not used in the activation of xenobiotics. Tips Xenobiotics is a high yield topic in both biochemistry and pharmacology. 35. The protein rich in basic amino acids, which functions in the packaging of DNA in chromosomes, is: A. Histone. B. Collagen. C. Hyaluronic acid-binding protein. D. Fibrinogen. Answer: A (Histone) Ref: Harper 25th edn, 412 Quality: Spotter Status: Repeat QTDF: Harper Discussion Histones are proteins found in the nuclei of all eukaryotic cells where they are complexed to DNA in chromatin and chromosomes. They are of relatively low molecular weight and are basic, having a very high arginine/lysine content. They are highly conserved and can be grouped into five major classes. Two copies of H2A, H2B, H3 and H4 bind to about 200 base pairs of DNA to form the repeating structure of chromatin, the nucleosome, with H1 binding to the linker sequence. They may act as non-specific repressors of gene transcription. Explanation A. Histones. B. Collagen is the protein substance of the white fibres (collagenous fibres) of skin, tendon, bone, cartilage and all other connective tissue, composed of molecules of tropocollagen, it is converted into gelatin by boiling. The word has originated from the Greek word Kolla = glue and gennan = to produce are the structural mole. C. Hyaluronic acid-binding protein occurs in pharyngeal epithelial cells. The capsular polysaccharide of Group A Streptococcus plays a role in colonisation of the pharynx by binding to CD44, a hyaluronic acid-binding protein expressed on human pharyngeal epithelial cells. Though this is not related to this question, I have given this because this may be asked in future. Hyaluronic acid, binding protein 2 is a serine protease. D. Fibrinogen is soluble plasma protein (340 kD, 46 nm long), composed of 6 peptide chains (2 each of A_, B_ and _) and present at about 2-3 mg/ml. Comments Questions from genetics are asked directly from Harper and are often taken from the blue headings given before each paragraph. Tips If you dont have time to read Harper thoroughly, read at least the blue headings given at the top of each paragraph. 36. Which of the following is not a post-transcriptional modification of RNA? A. Splicing. B. 5' capping. C. 3' polyadenylation. D. Glycosylation. Answer: D (Glycosylation) Ref: Harper 25th edn, 448; Lippincott 2nd edn, 385,386 Quality: Reader Status: Repeat QTDF: Lippincott Fig. 31.14, 31.15. Discussion Transcription, as we all know, is a step in the synthesis of mRNA from DNA. First heteronuclear RNA (hnRNA) is synthesised (which in nothing but a reverse or complimentary copy of the DNA from which it was synthesised) the sequence of which is exactly same as the other strand of DNA (except for the Uracil), and it undergoes few changes, (which are called as post-transcriptional modifications) to become an active mRNA. The changes are: 1. Additions 5 cappingAddition of 7 methyl guanosine capping to Initiate translation Protection of 5 end from attack of 5-3 exonuclease 3 cappingAddition of poly a tail or 3 polyadenylation 2. SplicingRemoval of introns and Merging the exons Explanation A. Splicing occurs. 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Tips Please be thorough with acid-base and electrolyte balance. 44. The following separation technique depends on the molecular size of the protein: A. Chromatography on a carboxymethyl (CM) cellulose column. B. Iso-electric focusing. C. Gel filtration chromatography. D. Chromatography on a diethylaminoethly (DEAE) cellulose column. Answer: C (Gel filtration chromatography) Ref: Harper 25th edn, 58, 42; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde, 73 and 74 Quality: Spotter Status: Repeat QTDF: Harper Discussion Chromatography is a broad range of physical methods used to separate and or to analyse complex mixtures. The components to be separated are distributed between two phases: A stationary phase bed and a mobile phase which percolates through the stationary bed. Chromatography Basis Principle Procedure Gel filtration Size Columns having pores, Unknown Molecular calibrated with proteins protein identified Weight of known size (mol wt) by elution Iso-electric Iso electric Each protein is Electricity is used focussing pH electrically neutral at a and hence this is particular pH electrophoresis Ion Exchange Electrostatic The resins used See the Tips chromatography Attraction are carboxy methyl between a cellulose or Diethyl charged aminoethyl cellulose molecule called CM and and DEAE. opposite charged molecule on the resins Explanation A. Chromatography on a carboxymethyl (CM) cellulose column is ion exchange chromatography. B. Iso-electric focusing is electrophoresis. C. Gel filtration chromatography uses the molecular weight or size of the molecules. D. Chromatography on a diethylaminoethly (DEAE) cellulose column is ion exchange chromatography. Comments Types of chromatography: Adsorption Chromatography Adsorption chromatography is probably one of the oldest types of chromatography around. It utilises a mobile liquid or gaseous phase that is adsorbed onto the surface of a stationary solid phase. The equilibriation between the mobile and stationary phase accounts for the separation of different solutes. Partition Chromatography This form of chromatography is based on a thin film formed on the surface of a solid support by a liquid stationary phase. Solute equilibriates between the mobile phase and the stationary liquid. Ion Exchange Chromatography In this type of chromatography, the use of a resin (the stationary solid phase) is used to covalently attach anions or cations onto it. Solute ions of the opposite charge in the mobile liquid phase are attracted to the resin by electrostatic forces. Molecular Exclusion Chromatography Also known as gel permeation or gel filtration, this type of chromatography lacks an attractive interaction between the stationary phase and solute. The liquid or gaseous phase passes through a porous gel which separates the molecules according to its size. The pores are normally small and exclude the larger solute molecules, but allow smaller molecules to enter the gel, causing them to flow through a larger volume. This causes the larger molecules to pass through the column at a faster rate than the smaller ones. Affinity Chromatography This is the most selective type of chromatography employed. It utilises the specific interaction between one kind of solute molecule and a second molecule that is immobilised on a stationary phase. For example, the immobilised molecule may be an antibody to some specific protein. When solute containing a mixture of proteins are passed by this molecule, only the specific protein is reacted to this antibody, binding it to the stationary phase. This protein is later extracted by changing the ionic strength or pH. Tips Another topic that is favourite for examiners is electrophoresis 45. The substances present in the gallbladder stones or the kidney stones can be best identified by the following technique: A. Fluorescence spectroscopy. B. Electron microscopy. C. Nuclear magnetic resonance. ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l )H,@ l IH,@ l ,@ l \X1X\X deposition of material on the pE,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l /,@ l ,@ l hXXXX,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l .,@ l LhIh,@ l ,@ l XXXX ,@ l u,@ l ,@ l h9Q<8h ,@ l XmXXGX ,@ l ,@ l ,@ l ,@ l :hXX$XX,@ l ,@ l _,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l S,@ l P,@ l hdhLhEh]hjh?,@ l ,@ l EXgXXhX ,@ l i,@ l ,@ l K,@ l ,@ l 3hXXXX,@ l ,@ l k,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l There are five major classes of haemoglobinopathies: 1. Structural haemoglobinopathies: occur when mutations alter the amino acid sequence of a globin chain, altering the physiologic properties of the variant haemoglobins and producing the characteristic clinical abnormalities. The variant haemoglobins relevant to this chapter: polymerise abnormally, as in sickle cell anaemia, or exhibit altered solubility or oxygen-binding affinity. 2. Thalassaemia syndromes arise from mutations that: impair production or translation of globin mRNA: leading to deficient globin chain biosynthesis. Clinical abnormalities are attributable: to the inadequate supply of hemoglobin and the imbalances in the production of individual globin chains, leading to premature destruction of erythroblasts and red cells. 3. Thalassaemic haemoglobin variants combine features of thalassaemia (e.g., abnormal globin biosynthesis) and of structural haemoglobinopathies (e.g., an abnormal amino acid sequence). 4. Hereditary persistence of foetal haemoglobin (HPFH) is characterised by: synthesis of high levels of foetal haemoglobin in adult life. 5. Acquired haemoglobinopathies include modifications of the haemoglobin molecule by toxins (e.g., acquired methaemoglobinemia), and abnormal haemoglobin synthesis (e.g., high levels of HbF production in preleukaemia and a-thalassaemia in myeloproliferative disorders). Few details about HbS Haematology Normal in the heterozygote; haemolytic anaemia of varying degrees in homozygotes; painful vaso-occlusive episodes; leg ulcers; jaundice, stroke, congestive heart failure; meningitis; lowered resistance to salmonella and pneumococcus organisms Lectrophoresis HbS and HbA can readily be separated at both alkaline and acidic pH Chromatography HbS and HbA separate by both cation and anion exchange chromatography Structure Tryptic digestion; fingerprinting; cation studies exchange chromatography; amino acid analysis; sequencing; HPLC DNA Analyses A GAG->GTG mutation at codon 6 functions Normal studies Stability Normal; in the deoxy form HbS precipitates, particularly at higher salt concentrations Occurrence Heterozygotes and homozygotes found in many ethnic groups but predominantly in the Black race and in some Indian tribes Other HbS is the most studied Hb variant Information Explanation A. An abnormality in porphyrin part of haemoglobin will lead to porphyrias. B. Replacement of glutamate by valine in B-chain of HbA is the cause of HbS. C. A nonsense mutation in the B-chain of HbA will lead to chain termination. D. Substitution of valine by glutamate in the alpha chain of HbA will lead Hb Pavie in which valine in position 135 is replaced by glutamate. And the following table gives the haemoglobin variants caused by the substitution of valine by glutamate in the beta chain of HbA Hb Position Original AA Replaced Original Mutation AA Sequence Trollhttan 20(B2) Val Glu GTG GAG Cagliari 60(E4) Val Glu GTG GAG M-Milwaukee-I 67(E11) Val Glu GTG GAG Mainz 98(FG5) Val Glu GTG GAG New York 113(G15) Val Glu GTG GAG Hofu 126(H4) Val Glu GTG GAG North Shore 134(H12) Val Glu GTG GAG Comments Since most of the details for this are already in all the books, let us see few facts which are likely to be asked in future exams. The types and number of haemoglobin variants are: Type Number Alpha chain variants 199 Beta chain variants 335 Gamma chain variants (G-gamma = 38; A-gamma = 20; unknown = 3; special = 7) 68 Delta chain variants 28 Variants with two amino acid replacements (alpha = 1; beta = 17) 18 Variants with hybrid chains 10 Variants with elongated chains (at the C-terminus = 9; at the N-terminus = 4) 13 Variants with deletions (15); with insertions (4); with deletions and insertions (3) 22 Total 693 Few haemoglobins where the substitution occurs in position 6 in beta chain: Hb Position Original Replaced Original Mutation AA AA Sequence S 6(A3) Glu Val GAG GTG C 6(A3) Glu Lys GAG AAG G-Makassar 6(A3) Glu Ala GAG GCG Machida 6(A3) Glu Gln GAG CAG One more alpha chain disorder which is: Ann Arbor 80(F1) Leu->Arg (CTG > CGG) Tips Refer Table 106 1 in Harrison 15th Edition and the relevant chapters in Nelson. For a complete list visit www.rxpgonline.com, www.targetpg.com 31. Decreased glycolytic activity impairs oxygen transport by haemoglobin due to: A. Reduced energy production. B. Decreased production of 2,3-bisphosphoglycerate. C. Reduced synthesis of haemoglobin. D. Low levels of oxygen. Answer: B (Decreased production of 2,3-bisphosphoglycerate) Ref: Harper 25th,70, 195; Lippincott 2nd edn, 31,92 Figure 7.8; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde, 258 Quality: Reader Status: Thinker QTDF: Harper Discussion 2,3-DPG (2,-3-biphosphoglycerate) is produced by the alternate pathway and when the glycolytic activity decreases, the amount of 2,-3 DPG decreases and we all know that 2, 3-DPG releases oxygen from Hb to the tissues. Thus when there is low amount of 2, 3-DPG, there is shift to left, that is oxygen is bound tightly to the Hb and as a result there is decreased availability of oxygen to the tissues. Explanation A. Oxygen delivery is not related to energy production. B. Decreased production of 2,3-bisphosphoglycerate leads to shift to left. C. Decreased glycolytic activity does not reduce synthesis of haemoglobin all of a sudden. D. Low levels of oxygen leads to decreased glycolytic activity (of course) but not the opposite as given in this choice. Comments XJ`J (Desired HCO3 Measured HCO3) Replacement in hour = 0.25 Body Weight (Desired HCO3 Measured HCO3) = 0.25 50 (25-5) = 250 mEq Comments The base excess is a calculated figure which provides an estimate of the metabolic component of the acid-base balance. The base excess is defined as the amount of H+ ions that would be required to return the pH of the blood to 7.35 if the PCO2 were adjusted to normal. Because the base excess is a calculated (not a measured) value, it may be inaccurate and misleading. Despite these problems, it is important to understand the concept. 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In short, more questions (75%) come from the topics you allot, 10% of your total time for Biochem. Does this tell something???? 28. The conversion of an optically pure isomer (enantiomer) into a mixture of equal amounts of both dextro and levo forms is called: A. Polymerisation. B. Stereoisomerisation. C. Racemisation. D. Fractionation. Answer: C (Racemisation) Ref: Harper 25th edn, 150 Quality: Spotter Status: New QTDF: Harper Discussion Any optically active compounds may occur as: D-isomer is dextro isomer which rotates the polarised light to the right L-isomer is levo isomer which rotates the polarised light to the left When equal amounts of D and L forms are present, the mixture is called Racemic or DL mixture. (A mixture of equal amounts of the two stereoisomers of an optically active substance). In this case, the polarised light is not rotated as the actions of the D and L form cancel each other. Explanation A. Polymerisation is the act or process of changing to a polymeric form and the condition resulting from such change. For example, monovinyl chloride to polyvinyl chloride or glucose to starch or cellulose or glycogen. B. Stereoisomerisation is the process of interchanging compounds whose molecules have the same number and kind of atoms and the same atomic arrangement, but differ in their spatial relationships. C. Racemisation is partial conversion of one enantiomorph into another (as an l-amino acid to the corresponding d-amino acid), so that the specific optical rotation is decreased, or even reduced to zero, in the resulting mixture. D. Fractionation is a term used to describe any method for separating and purifying biological molecules. Commonly used in separation of cell component (Another use is the seperation of petrol, diesel, etc. from crude oil). Comments About polarisation, crystals of gout are negatively birefringent and that of pseudogout are positively birefringent. Tips Isomers: Due to a asymmetrical carbon: D-glucose (OH group on right) and L-glucose (Capital D and L). Optical isomers are enantiomeres: Dextro rotatory or D or + and levo rotatory or l or negative (note the small d and l). Anomers: They are different in carbon 1 : alpha D-Glucose and beta D-Glucose Epimers: The sugars are different with respect to orientatioin of one carbon atom. 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Anaemia, obstructive lung disease, cystic fibrosis, and congenital heart disease are all accompanied by increases in 2,3-DPG. When more oxygen is required because of increased metabolism, such as in hyperthyroidism, more 2,3-DPG is produced. Decreased 2,3-DPG results from an inherited lack of the red blood cell enzymes 2,3-DPG mutase and 2,3-DPG phosphatase. These enzymes are needed to make 2,3-DPG. Without 2,3-DPG to control the movement of oxygen to its tissues, the body responds by making more red blood cells, leading to polycythaemia (erythrocytosis). The outside membrane of the cell is weakened, causing it to have an irregular shape and burst, or haemolyse, easily. This condition leads to nonspherocytic haemolytic anaemia. 2,3-DPG levels are important in large blood transfusions, because stored blood quickly loses 2,3-DPG and its ability to deliver oxygen. After transfusion, the red cells rebuild the 2,3-DPG, but it takes about 24 hours to regain a normal level of 2,3-DPG and haemoglobin function. Tips Read the alternate pathways of glucose metabolism 32. An enzyme involved in the catabolism of fructose to pyruvate in the liver is: A. Glyceraldehyde-3-phosphate dehydrogenase. B. Phosphoglucomutase. C. Lactate-dehydrogenase. D. Glucokinase. Answer: A (Glyceraldehyde-3-phosphate dehydrogenase). Ref: Harper 25th, 192,225 Figure 19.4; Lippincott 2nd edn, 128,129; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 288 Quality: Reader Status: Repeat QTDF: Harper Discussion Glyceraldehyde-3-phosphate dehydrogenase is an enzyme that is involved in the catabolism of fructose to pyruvate. Refer to the figure 19.4 Harper 25th edition for more details. Explanation A. Glyceraldehyde-3-phosphate dehydrogenase catalyses the transformation of glyceraldehyde-3-Phosphate to 1,3-bisphosphoglycerate which is eventually converted to pyruvate. This enzyme is a tetramer. B. Phosphoglucomutase converts alpha-D-glucose-6-phospate to alpha-D-glucose-1-phospate. In short, it leads to the uronic acid pathway. It is also called as glucose phosphomutase. And the maximum activity is only obtained in the presence of alpha-D-glucose 1,6-bisphosphate. This bisphosphate is an intermediate in the reaction, being formed by transfer of a phosphate residue from the enzyme to the substrate, but the dissociation of bisphosphate from the enzyme complex is much slower than the overall isomerisation. Also, more slowly, catalyses the interconversion of 1-phosphate and 6-phosphate isomers of many other alpha-D-hexoses, and the interconversion of alpha-D-ribose-1-phosphate and 5-phosphate. Also, called EC 5.4.2.2 Formerly called EC 2.7.5.1. C. Lactate dehydrogenase is an oxidoreductase which catalyzes the conversion of lactate to pyruvate. It consists of 4 subunits which may be of 2 different types = M and H (muscle and heart formerly known as A and B respectively). 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The molecular weight of all isoenzymes is ca. 140 kDa. L(+)-lactate dehydrogenase is specific for L(+)-lactate and does not react with D()-lactate. LDH is used in coupled enzyme assays, for example, in the determination of ATPase, myokinase and pyruvatekinase. It may also be used in the determination of lactate (5), pyruvate (6) and various other metabolites. D. Glucokinase [Hexokinase D (EC 2.7.1.1)] is the characteristic isoenzyme of hexokinase in the liver, and it is responsible for glucose phosphorylation in hepatocytes. Its importance derives from the fact that glucose uptake by the liver is an essential physiological process, crucial for glucose homeostasis. And it can phosphorylate glucose only and cannot be involved in fructose metabolism. Comments This enzyme is also called NAD-dependent glyceraldehyde-3-phosphate dehydrogenase and triosephosphate dehydrogenase and it also acts very slowly on D-glyceraldehyde and some other aldehydes. Thiols can replace phosphate. Tips The reaction catalysed by glyceraldehyde-3-phosphate dehydrogenase catalyses is an reaction important in the generation of ATP. Make a note of all the reactions in various pathways in which high energy phosphates and NAD/NADP are involved. 33. B-oxidation of odd-chain fatty acids produces: A. Succinyl CoA. B. Propionyl CoA. C. Acetyl CoA. D. Malonyl CoA. Answer: B (Propionyl CoA) Ref: Harper 25th, 240; Lippincott 2nd edn, 183, 184 Figure 17.17; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 381; Chaterjee 2nd edn, 542. MCQ given at the end of the chapter 20 Quality: SpotterFirst MBBS Status: Repeat QTDF: ChaterjeeMCQ given at the end of the chapter Discussion Beta oxidarion of even chain fatty acids produce acetly CoA and that of odd chain fatty acids producs acetyl CoA and one propionyl CoA. Explanation A. Succinyl CoA is not produced. B. One molecule of propionyl CoA is produced. C. Acetyl CoA also is produced but it is produced in odd as well as even chain chain fatty acids. D. Malonyl CoA is not produced. Comments A question that is repeated for a long period of time. Tips Go through the MCQs given at the end of each chapters in Chatterjee. Questions are being asked from that repeatedly. 34. The main enzyme responsible for activation of xenobiotics is: A. Cytochrome P-450 B. Glutathione S-transferase. C. NADPH cytochrome P-450-reductase. D. Glucuronyl transferase. Answer: A (Cytochrome P-450) Ref: Harper 24th edn, Chapter 61 Metabolism of Xenobiotics, 750; Harper 25th edn, 781; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 487 Quality: Thinker !!! Status: New QTDF: Harper Discussion A xenobiotic (Gk xenos stranger) is a compound that is foreign to the body, the principal classes of xenobiotics of medical relevance are drugs, chemical carcinogens, and various compounds that have found their way into our environment. It is convenient to consider the metabolim of xenobiotics in two phases. In phase 1, the major reaction involved is hydroxylation, catalysed by members of a class of enzymes referred to as mono-oxygenases or cytochrome P-450s. Hydroxylation may terminate the action of a drug, though this is not always the case. In addition to hydroxylation, these enzymes catalyse an astonishingly a wide range of reactions, including those involving deamination, dehalogenation, desulphuration, epoxidation, peroxygenation, and reduction. Reactions involving hydolysis (e.g., catalysed by esterases) and certain other non-P450-catalysed reactions also occur in phase 1. In phase 2, the hydroxylated or other compounds produced in phase 1 are converted by specific enzymes to various polar metabolites by conjugation with glucuronic acid, sulphate, acetate, glutathione or certain amino acids or methylation. Oxidation by mono-oxygenases proceed as the following reaction, catalysed by cytochrome P-450 (and in the process of which cytochrome P-450 turn into oxidised P-450. Reduced cytochrome P-450 + RH + O2 R-OH + H2O + Oxidised Cytochrome P-450 And here in this reaction, which is the reaction responsible for activation of xenobiotics (asked in the question) cytochrome P-450 is involved and then there is another reaction in which NADPH, not NADH is involved. It is in the reaction mechanism of cytochrome P-450. The enzyme that uses NADPH to yield the reduced cytochrome P-450, shown in the left hand side of the above equation, is called NADPH - cytochrome P-450 reductase. Electrons, are transferred from NADPH to NADPH-cytochrome P-450 reductase and then to cytochrome P-450. 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The buffering capacity of a buffer is maximum at pH equal to: A. 0.5 pKa. B. pKa C. pKa+1 D. 2 pKa Answer: B (pKa) Ref: Harper 25th edn, 23 Figures 3.6, 3.7 Quality: Reader Status: Repeat QTDF: Harper Discussion pKa of a substance is the pH at which it is 50% ionised. By buffers, we mean those substances that resist changes in pH when an acid or base is added into the medium. Buffering is a tendency of a substance which is not equal at all ranges of pH. The usual buffers are solutions of weak acids/bases and their salts. Explanation A. pH of 0.5 pKa means pH < pKa, that is, at an acidic pH the capacity of the buffer to resist change when an acid is added is greatly reduced. B. The buffering capacity is maximum at pKa. C. pKa + 1 is alkaline pH and the capacity of the buffer to resist change when an alkali is added is greatly reduced. D. pH of 2 pKa means pH > pKa, that is, at an alkaline pH the capacity of the buffer to resist change when an alkali is added is greatly reduced. Comments Please note that answers C and D are themselves absurd. For example, in choice C, there cannot be a pH when pKa is more than 13 and for choice D there will be no pH if the pKa is more than 7!!! (how can pH be more than 14). Tips Often in Objective exams, few choices will stand out if we carefully analyse, even though when we dont know the correct answer !!!. The hurdle is the limited time (about 40 seconds/question in AIPG). To improve your speed practise the mock tests at www.rxpg.com and www.rxpgonline.com 30. The primary defect which leads to sickle cell anaemia is: A. An abnormality in porphyrin part of haemoglobin. B. Replacement of glutamate by valine in B-chain of HbA. C. A nonsense mutation in the B-chain of HbA. D. Substitution of valine by glutamate in the a-chain of Hba. Answer: B (Replacement of glutamate by valine in B-chain of HbA) Ref: Harper 25th edn, 71; Lippincott 2nd edn, 35; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde Page 134 Quality: First MBBS Status: Repeat from the time MCQs are asked !!!! QTDF: All books have this (Physio, Biochem, Path, Medicine, Paed) Discussion ,(xKdd9X(xKdddLX(xKddd^X(xKdddzX(xKddd ,(xKddP (xKdd (xKdd (xKdd ,(xKdd2 (xKdd (xKddSh<(xKddd.X(xKdddX(xKdddX(xKdddX(xKddd ,(xKdd1 ,(xKdd (xKdd (xKdd (xKdd (xKdd (xKdd (xKdd (xKdd (xKdd ,(xKdd (xKdd ,(xKddX(xKdddX(xKdddFh<(xKddd(xKddd(xKddd(xKddd(xKddd(xKddd(xKdddX(xKddd ,(xKdd (xKdd ,(xKdd (xKdd (xKdd4h<(xKdddX(xKdddX(xKdddX(xKdddlGáD*w-kDLD*w-kH!0߸pP0PP -0񄋋D_-kD'|x%A\Pb0񄋋D_-kD'|x%A\Pb0߸pP0PP .Gd<Gd22GdIn acidosis, the pump mechanism is altered and the intracellular potassium wanders out of its home into the ECF During tissue injury, due to the loss of cell membrane integrity, hyperkalaemia occurs Hypokalaemia (Increased potassium inside cells): a. Insulin b. Aldosterone c. Alkalosis d. Beta adrenergic drugs (Salbutamol) Hyperkalaemia (Decreased potassium outside cells): a. Diabetes b. Addison's c. Acidosis d. Beta blockersPropanolol e. Cell injury f. Exercises Explanation A. Most of potassium is intracellular. B. Three quarter of the total body potassium is found in skeletal muscle. C. Intracellular potassium is released into extracellular space in response to severe injury remember that rhabdomyolysis leads to hyperkalaemia. D. Acidosis leads to movement of potassium from intracellular to extracellular fluid compartment. Comments There will be no question paper without a question on sodium, potassium, calcium, acidosis, alkalosis (Acid and Electrolyte Balance), etc. In any question paper, you can expect 5% of questions on this. Tips Prepare well. Read the relevant chapters in all books, For certain topics, it is enough if you read any one of the book, but for this you have to read all the concerned books. 41. Causes of metabolic alkalosis include all the following, except: A. Mineralocorticoid deficiency. B. Bartters syndrome. C. Thiazide diuretic therapy. D. Recurrent vomiting. Answer: A (Mineralocorticoid deficiency) Ref: Harrison 15th edn, Chapter 50, Table 50.4 and 288 Quality: Reader Status: Repeat QTDF: Harrison Discussion Metabolic alkalosis occurs as a result of net gain of [HCO3] or loss of nonvolatile acid (usually HCl by vomiting) from the extracellular fluid. Since it is unusual for alkali to be added to the body, the disorder involves a generative stage, in which the loss of acid usually causes alkalosis, and a maintenance stage, in which the kidneys fail to compensate by excreting HCO3 because of volume contraction, a low GFR, or depletion of Cl or K. Under normal circumstances, the kidneys have an impressive capacity to excrete HCO3. Continuation of metabolic alkalosis represents a failure of the kidneys to eliminate HCO3 in the usual manner. For HCO3 to be added to the extracellular fluid, it must be administered exogenously or synthesised endogenously, in part or entirely by the kidneys. The kidneys will retain, rather than excrete, the excess alkali and maintain the alkalosis if: 1. volume deficiency, chloride deficiency, and K+ deficiency exist in combination with a reduced GFR, which augments distal tubule H+ secretion; or 2. hypokalaemia exists because of autonomous hyperaldosteronism. In the first example, alkalosis is corrected by administration of NaCl and KCl, while in the latter it is necessary to repair the alkalosis by pharmacologic or surgical intervention, not with saline administration. Explanation A. In patients with decreased mineralocorticoid production or action, disorders of aldosterone biosynthesis or action are associated with high renin levels, salt wasting, and hyperkalaemia. The aldosterone levels may be low or elevated. In patients with a deficiency in aldosterone biosynthesis, the transformation of corticosterone into aldosterone is impaired, owing to a mutation in the aldosterone synthase (CYP11B2) gene. These patients have low to absent aldosterone secretion, elevated plasma renin levels, and elevated levels of the intermediates of aldosterone biosynthesis (corticosterone and 18-hydroxycorticosterone). Pseudohypoaldosteronism type I (PHA-I) is an autosomal recessive disorder that is seen in the neonatal period and is characterised by salt wasting, hypotension, hyperkalemia, and high renin and aldosterone levels. In contrast to the gain-of-function mutations in the epithelial sodium channel (ENaC) in Liddles syndrome, mutations in PHA-I result in loss of ENaC function. B. GdZB. 5' capping is the first process and 7-methyl guanosine is attached from the back to stabilise the mRNA. C. 3' polyadenylation occurs when about 40 to 200 adenine nucleosides are attached to the 3 end of the mRNA and this stabilises and facilitates the exit from the nucleus. This tail is later shortened. D. Glycosylation does not occur as a post-translational modification. Comments DNA, RNA and protein synthesis are potential areas for MCQs and MCQs from these areas are often repeated even from pharmacology. Tips SLE is due to antibodies against SnRnPs. SnRNA, in association with proteins forming small nuclear ribonucleo protein particles (SnRNPs) facilitated the splicing of some exons segments by forming base pairs with each end of the untron. 37. The collagen triple helix structure is not found in: A. Cytoplasm. B. Golgi apparatus. C. Lumen of endoplasmic reticulum. D. Intracellular vesicles. Answer: A (Cytoplasm) Ref: Harper 25th edn, 697 and Table 57.3; Lippincott 2nd edn, 40 to 42 and Figure 3.26 Quality: Spotter Status: New QTDF: Lippincott Discussion Collagen is a protein that functions outside the cell and is a structural protein. The various stages in the synthesis of collagen can be summarised as follows: 1. Genes for pro-a1 and pro-a2 chains are transcribed into mRNAs 2. mRNA is translated into pre pro a-polypeptide chains that are extruded into the endoplasmic reticulum where signal sequence is removed 3. Selected proline and lysine residues are hydroxylated 4. Selected lysisne residues are glycosylated with glucose and galactose 5. Three pro alpha chains assemble. Intra and interchain disulphide bonds form at C terminal propeptide extension 6. Triple helix forms by zipper-like folding 7. Procollagen molecule secreted from Golgi vacuole into extracellular matrix through the vesicles 8. N terminal and C terminal propeptides cleaved by procollagen peptidases 9. Self-assembly of collagen molecules and subsequent cross linking 10. Oxidative deamination of epsilon amino group of lysyl and hydroxyl residues to aldehyde 11. Formation of intra- and interchain cross linkings as Schifff's bases and Aldol condensation products Steps 1 to 6 are intracellular and 8 to 11 are extracellular. Explanation A. The triple helix structure is not formed inside the cytoplasm. B. The triple helix structure is found inside Golgi apparatus from which they go to the vesicles. C. The triple helix structure is found inside lumen of endoplasmic reticulum from which they are send to the Golgi apparatus. D. The triple helix structure is found inside intracellular vesicles from where they are secreted out. Comments A new question from Lippincott (even though the answer is in Harper it is best explained in Lippincott). Tips Go through the figures in Lippincott (of at least the first few chapters). 38. The primary role of chaperones is to help in: A. Protein synthesis. B. 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In antenatal and classic Bartters syndrome, impaired Cl reabsorption in the thick ascending limb of the loop of Henle is the underlying defect. Inadequate Cl reabsorption causes volume depletion and activates the renin-angiotensin system. Distal delivery of NaCl and water are high in the presence of high aldosterone, promoting secretion of K+ and H+ ions. Prostaglandin overproduction is mediated by volume depletion, hypokalaemia, and high angiotensin II and kallikrein levels. Increased prostaglandin production contributes to the severity of disease by inducing resistance to the pressor effects of angiotensin II and reducing reabsorption in the thick ascending limb of the loop of Henle. Mutations in the bumetanide-sensitive Na:K:2Cl channel, the apical ATP-regulated K+ channel, and the basolateral Cl channel have been described in classic and antenatal Bartters. C. Thiazide diuretics increase urinary excretion of sodium and water by inhibiting sodium reabsorption in the early distal tubules. They increase the rate of delivery of tubular fluid and electrolytes to the distal sites of hydrogen and potassium ion secretion, while plasma volume contraction increases aldosterone production. The increased delivery and increase in aldosterone levels promote sodium reabsorption at the distal tubules, thus increasing the loss of potassium and hydrogen ions. And this leads to alkalosis. D. Recurrent vomiting leads to loss of acid and that leads to alkalosis. Comments Metabolic acidosis leads to hyperkalaemia as a result of cellular shifts in which H+ is exchanged for K+ or Na+. For each decrease in blood pH of 0.10, the plasma [K+] should rise by 0.6 mmol/L. This relationship is not invariable. Diabetic ketoacidosis, lactic acidosis, diarrhoea, and renal tubular acidosis (RTA) are often associated with potassium depletion because of urinary K+ wasting. Tips Read about RTA (Renal Tubular Acidosis) in Harrison. 42. Normal anion gap metabolic acidosis is caused by: A. Cholera. B. Starvation. C. Ethylene glycol poisoning. D. Lactic acidosis. Answer: A (Cholera) Ref: Harrison 15th edn, Chapter 50 and, 285 Quality: Reader Status: Repeat QTDF: Harrison Discussion There are four principal causes of a high-anion gap acidosis: 1. Lactic acidosis: a. Anaerobic metabolism predominatingshock/cardiopulmonary arrest b. Severe anaemia, c. Poisoning with CO or cyanide, 2. Ketoacidosis: a. Diabetes b. Alcoholics c. Starvation 3. Ingested toxins: a. Ethylene glycol b. Salicylates c. Methanol 4. Acute and chronic renal failure: Explanation A. In cholera the anion gap is normal. B. Starvation is a cause of increased anion gap acidosis C. Ethylene glycol poisoning is a cause of increased anion gap acidosis D. Lactic acidosis is a cause of increased anion gap acidosis. Comments All evaluations of acid-base disorders should include a simple calculation of the anion gap (AG); It represents those unmeasured anions in plasma (normally 10 to 12 mmol/L) It is calculated as follows: AG = Na+ (Cl + HCO3). The unmeasured anions include anionic proteins, phosphate, sulphate, and organic anions. When acid anions, such as acetoacetate and lactate, accumulate in extracellular fluid, the AG increases, causing a high-AG acidosis. An increase in the AG is most often due to an increase in unmeasured anions and less commonly is due to a decrease in unmeasured cations (calcium, magnesium, potassium). In addition, the AG may increase with an increase in anionic albumin, either because of increased albumin concentration or alkalosis, which alters albumin charge. A decrease in the AG can be due to: An increase in unmeasured cations; The addition to the blood of abnormal cations, such as lithium (lithium intoxication) or cationic immunoglobulins (plasma cell dyscrasias); A reduction in the major plasma anion albumin concentration (nephrotic syndrome); A decrease in the effective anionic charge on albumin by acidosis; or Hyperviscosity and severe hyperlipidaemia, which can lead to an underestimation of sodium and chloride concentrations. Tips Other conditions with normal anion gap are 1. Diarrhoea 2. Fistula 3. Ureterosigmoidostomy 4. Renal tubular acidosis 5. Ingestion of ammonium chloride 6. Mineralocorticoid deficiency. 43. A 50 kg man with severe metabolic acidosis has the following parameters: pH 7.05. pCO2 12 mmHg, pO2 108 mmHg. HCO3 5 mEq/L. base excess-30 mEq/L. The approximate quantity of sodium bicarbonate that he should receive in half hour is: A. 250 mEq. B. 350 mEq. C. 500 mEq. D. 750 mEq. Answer: A (250 mEq) Ref: Harrison 15th edn, Chapter 50 page 284 Quality: Reader Status: Repeat QTDF: Harrison Discussion Bicarbonate replacement Not usually recommended unless pH < 7.2 or HCO3- deficit is greater than 5 mEq/L. 8.4% NaHCO3 = 1 mEq/ml 1 gm of baking soda = 12 mEq of NaHCO3 Formula for bicarbonate replacement 0.3 (BW in kg) (HCO3- deficit) = mEq of NaHCO3 needed 0.3 assumes 30% of BW is extracellular fluid. If the figure of 0.3 is used, bicarbonate replacement can be given fairly rapidly (30 minutes to 1 hour). If figure of .5 is used bicarbonate replacement should be given over 12-24 hour period and half of it should be given in the first half hour. If you have a mixed respiratory/metabolic acidosis, important to address the respiratory acidosis first, before treating the metabolic acidosis. Explanation In our case replacement = 0.5 Body weight (Desired HCO3 Measured HCO3) Replacement in hour = 0.25 Body weight (Desired HCO3 Measured HCO3) = 0.25 50 (25-5) = 250 mEq Comments The base excess is a calculated figure which provides an estimate of the metabolic component of the acid-base balance. The base excess is defined as the amount of H+ ions that would be required to return the pH of the blood to 7.35 if the PCO2 were adjusted to normal. Because the base excess is a calculated (not a measured) value, it may be inaccurate and misleading. Despite these problems, it is important to understand the concept. 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Restriction fragment length polymorsphism is used for: A. Analysis of chromosome structure. B. DNA estimation. C. Synthesis of nucleic acid. D. Detecting proteins in a cell. Answer: B (DNA estimation) Ref: Harper, 25th edn, 500; Harrison 15th edn, Table 65-9; Lippincott 2nd edn, 415 and Figure 33.13 Quality: Spotter Status: Repeat QTDF: Harrison Discussion Methods Used for the Detection of Mutations Method Principle Type of Mutation Detected Cytogenic Unique visual appearance of Numerical or structural analysis various chromosomes abnormalities in chromosomes Fluorescent Hybridisation to chromo- Numerical or structural in situ somes with fluorescently abnormalities in hybridisation labelled probes chromosomes (FISH) Southern blot Hybridisation with genomic Large deletion, insertion, probe or cDNA probe after rearrangement, expansions of digestion of high molecular triplet repeat, amplification DNA Polymerase Amplification of DNA Expansion of triplet repeats, chain reaction segment variable number of tandem (PCR) repeats (VNTR), gene rearrangements, translocations; prepare DNA for other mutation methods Reverse Reverse transcription, Analysed expressed mRNA transcriptase amplification of DNA (cDNA) sequence; detect loss PCR (RT-PCR) segment absence or of expression reduction of mRNA transcription DNA Direct sequencing of Point mutations, small sequencing PCR products deletions and insertions Sequencing of DNA segments cloned into plasmid vectors Single-strand PCR of DNA segment: Point mutations, small conformational Mutations result in deletions and insertions polymorphism conformational change and (SSCP) altered mobility Contd... Contd... Denaturing PCR of DNA segment: Point mutations, small gradient gel Mutations result in deletions and insertions electrophoresis conformational change and (DGGE) altered mobility RNAse Cleavage of mismatch Point mutations, small cleavage between mutated and deletions and insertions wild-type sequence Restriction Detection of altered fragment restriction pattern of genomic length poly- DNA (Southern blot) or Point mutations, small morphism PCR products deletions and insertions (RFLP) Oligo- Hybridisation of PCR Point mutations, small nucleotide products to wild-type deletions and insertions specific or mutated oligonucleotides hybridisation immobilised on chips or (OSH) slides Microarrays Hybridisation of PCR Point mutations, small products to wild-type or deletions and insertions mutated oligonucleotides Protein Transcription/translation of Mutations leading to truncation cDNA isolated from tissue premature truncations test (PTT) sample Explanation A. Analysis of chromosome structure is by cytogenic analysis and fluorescent in situ hybridisation (FISH). B. DNA estimation is by restriction fragment length polymorphism (RFLP). C. Synthesis of nucleic acid is used in various other methods listed above. D. Detecting proteins in a cell is by Western Blot. Comments Strictly speaking, DNA is a chromosome and chromosome is a DNA and so there are few who might tell that the answer is A. But in case of any controversy, we always go by a standard book and here Harrison is taken as a standard. But remember that with RFLP we cannot see the chromosome which is needed if we want to analyse the structure. Tips Southern was the name of the person who invented Southern Blot, a process of determining the DNA structure. Then the process of determining RNA was called Northern Blot and the process of determining proteins Western Blot (DNA RNA Protein) What will be then Eastern Blot J 48. Serum total lactate dehydrogenase level will NOT be raised in: A. Muscle crush injury. B. Stroke. C. Myocardial infarction. D. Haemolysis. Answer: B (Stroke) Ref: Textbook of Medical Biochemistry 2nd edn, Chaterjee and Shinde, 264 and 967; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde, 572 Quality: Spotter Status: Repeat QTDF: Chaterjee Discussion LDH or lactate dehydrogenase is an enzyme involved in glycolysis and it converts pyruvate to lactate under anaerobic conditions. In the presence of oxygen lactic acid can be oxidized to pyruvic acid again. The characteristic features of this reaction are: Reversible Same enzyme Same co-enzyme Oxidation reduction LDH is found more in the following cells: Cardiac muscle Skeletal muscle Liver Kidney RBC Normal value of LDH is 60 to 250 IU/L (120 500 units/ml by karmen spectroscopic method) It is elevated in acute myocardial infarction, carcinomatosis, acute leukaemias, granulocytic leukaemia, pulmonary infarction, renal necrosis and muscle disease. Less pronounced LDH increases are seen in inflammatory hepatic disorders. Explanation A. Muscle crush injury is associated with elevated levels. B. Stroke is not associated with elevated levels. C. Myocardial infarction there is elevated LDH level. The peak rise in serum LDH is roughly proportional to the extent of the injury to the myocardial tissue The elevated levels may persist for more than a week after CPK and SGOT levels have returned to normal Serum LDH level > 1500 IU/L in AMI suggests grave prognosis D. Red blood cells are rich in LDH, hence in haemolysis LDH levels are elevated. Particular care has to be taken while measuring LDH levels to ascertain that there is no haemolysis in the sample. Comments There are five isoenzymes fo LDH. The active LDH molecule is a tetramer of molecular weight 130000 having 2 M and 2 H each having mol wt of 34,000 Daltons. Only the tetrameric forms are enzymatically active: Iso- Subunits Mol Found Starch Electrophoresis enzyme Formula in LDH 1 HHHH H4 Cardiac Moves far towards anode, b/w albumin and alpha-1-globulin LDH 2 HHHM H3M alpha-2-globulin LDH 3 HHMM H2M2 Beta region LDH 4 HMMM HM3 Fast gamma LDH 4 MMMM M4 Liver Behind gamma globulins Tips Read the other enzymes. etions and insertions wild-type sequence Restriction y8Y0񄋋D_-kD'|x%A\Pb '\P-kDlGáD*w-kDLD*w-kH!0Pp0PP z '\P-kDlGáD*w-kDLD*w-k '\P-kDlGáD*w-kDLD*w-kH!0Pp0PP {0񄋋D_-kD'|x%A\Pb '\P-kDlGáD*w-kDLD*w-kH!0߸pP0PP } '\P-kDlGáD*w-kDLD*w-k '\P-kDlGáD*w-kDLD*w-kH!0߸pP0PP ~.50񄋋D_-kD'|x%A\Pb '\P-kDlGáD*w-kDLD*w-kH!0߸pP0PP am ChKcamCh9camChPc*C, camChcamChcamChcamChHcamChec7#,#C c7+,3#>#:# C@B, Cca:4+XhCXcaN/CX CxcaD3CXcasm3#5Cxcasm5#2C:casm2# Cca05#Xh:#Xh6#XhCX @B, Cz:#:#<# C CdCcp cpDcpcplocp CpcpcpDcpCplvCdCcpcp<cpDcpcpl#p cpcp<cpDcpcplCp ocpcp<cpDcp*cplcp Cpcpcp<cpDcp[cpl cp Cpcpcp<cpDcp*cpl cp Cpcc<cDcclGc ?CCdC@B,CrClGáD*wn the proper folding of other proteins without themselves being a compound of latter. They stabilise unfolded or partially folded intermediates, allowing them to fold properly and prevent inappropriate interactions. Few substances that can be called as chaperones include: 1. Calreticulin 2. HSP 90A (Heat shock proteins) 3. Synuclein alpha 4. Lectin mannose binding 1 5. DNA fragmentation factor 45 6. Tubulin specific chaperone C 7. Arylhydrocarbon-interacting receptor protein-like 1 8. Tubulin specific chaperone D 9. Unactive progesterone receptor 23KD (Yeah, That is its nameit is not printing mistake) Explanation A. Protein synthesis is a big process that involves a whole lot of enzymes. B. Protein degradation is by the digestive enzymes. C. Protein denaturation is usually by heat, organic solvents, mechanical mixing, strong acids or bases, detergents and ions of heavy metals such as lead and mercury. D. Protein folding is by chaperones. Comments Few more points about chaperones 1. Present in many species 2. Few are induced by conditions which cause unfolding of newly synthesised proteins 3. Bind predominantly to hydrophobic regions 4. Act as editing mechanisms or quality control 5. Most have ATPase activity 6. Found in: a. Cytosol b. Mitochondria c. Lumen of endoplasmic reticulum. Tips Read the enzymes involved in human as well as bacterial protein synthesis with relevance to pharmacology (Anti- microbial and chemotherapeutic agent). 39. Which of the following is present intracellularly in muscle cells: A. Insulin. B. Corticosteroid. C. Epinephrine. D. Glucagon. Answer: B (Corticosteroid) Since all are present inside the cell, we read the question as for which of the following, the receptor is present inside the cell and then the answer will be. Ref: KDT 4th edn, 240,241; KDT 5th edn, 214; Ganong Quality: Spotter with a small mistake Status: Repeat QTDF: Ganong Discussion 1. Receptors for most of the hormones are in the cell membrane: Act by camp Adrenaline, glucagon, TSH, FSH, LH, PTH, ACTH, some hypothalamic releasing factors Act by IP3/DAG: Vasopressin Oxytocin Act by enzyme activation: Tyrosine LinaseInsulin Receptors of few are inside the cell: In cytoplasm: Steroids other than estrogen In nucleus: Estrogen (Refer Ganong Chapter 1 and the MCQs given at the back of the book in Chapter 1) Thyroid hormones T3, T4 Explanation A. Insulin acts by transmembrane enzyme activating receptor. B. Corticosteroid has intracellular receptors. C. Epinephrine acts by membrance receptor. D. Glucagon acts by membrane receptors. Comments After the exam, the answer for this question was not debated and it was found that most had attempted even though they knew that the question is wrong !!! Tips If you feel that there is a mistake in the question and ONLY one of the choices could be the right choice if the question was corrected then go for it. 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wasting, hypotension, hyperkalemia, and high renin and aldosterone levels. In contrast to the gain-of-function mutations in the epithelial sodium channel (ENaC) in Liddles syndrome, mutations in PHA-I result in loss of ENaC function. B. @Bc)Increasing the amount of 2,3-DPG is the bodys primary way of responding to a lack of oxygen. Anaemia, obstructive lung disease, cystic fibrosis, and congenital heart disease are all accompanied by increases in 2,3-DPG. When more oxygen is required because of increased metabolism, such as in hyperthyroidism, more 2,3-DPG is produced. Decreased 2,3-DPG results from an inherited lack of the red blood cell enzymes 2,3-DPG mutase and 2,3-DPG phosphatase. These enzymes are needed to make 2,3-DPG. Without 2,3-DPG to control the movement of oxygen to its tissues, the body responds by making more red blood cells, leading to polycythaemia (erythrocytosis). The outside membrane of the cell is weakened, causing it to have an irregular shape and burst, or haemolyse, easily. This condition leads to nonspherocytic haemolytic anaemia. 2,3-DPG levels are important in large blood transfusions, because stored blood quickly loses 2,3-DPG and its ability to deliver oxygen. After transfusion, the red cells rebuild the 2,3-DPG, but it takes about 24 hours to regain a normal level of 2,3-DPG and haemoglobin function. Tips Read the alternate pathways of glucose metabolism 32. An enzyme involved in the catabolism of fructose to pyruvate in the liver is: A. Glyceraldehyde-3-phosphate dehydrogenase. B. Phosphoglucomutase. C. Lactate-dehydrogenase. D. Glucokinase. Answer: A (Glyceraldehyde-3-phosphate dehydrogenase). Ref: Harper 25th, 192,225 Figure 19.4; Lippincott 2nd edn, 128,129; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 288 Quality: Reader Status: Repeat QTDF: Harper Discussion Glyceraldehyde-3-phosphate dehydrogenase is an enzyme that is involved in the catabolism of fructose to pyruvate. Refer to the figure 19.4 Harper 25th edition for more details. Explanation A. Glyceraldehyde-3-phosphate dehydrogenase catalyses the transformation of glyceraldehyde-3-Phosphate to 1,3-bisphosphoglycerate which is eventually converted to pyruvate. This enzyme is a tetramer. B. Phosphoglucomutase converts alpha-D-glucose-6-phospate to alpha-D-glucose-1-phospate. In short, it leads to the uronic acid pathway. It is also called as glucose phosphomutase. And the maximum activity is only obtained in the presence of alpha-D-glucose 1,6-bisphosphate. This bisphosphate is an intermediate in the reaction, being formed by transfer of a phosphate residue from the enzyme to the substrate, but the dissociation of bisphosphate from the enzyme complex is much slower than the overall isomerisation. Also, more slowly, catalyses the interconversion of 1-phosphate and 6-phosphate isomers of many other alpha-D-hexoses, and the interconversion of alpha-D-ribose-1-phosphate and 5-phosphate. Also, called EC 5.4.2.2 Formerly called EC 2.7.5.1. C. Lactate dehydrogenase is an oxidoreductase which catalyzesthe conversion of lactate to pyruvate. It consists of 4 subunits which may be of 2 different types = M and H (muscle and heart formerly known as A and B respectively). 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They stabilise unfolded or partially folded intermediates, allowing them to fold properly and prevent inappropriate interactions. Few substances that can be called as chaperones include: 1. Calreticulin 2. HSP 90A (Heat shock proteins) 3. Synuclein alpha 4. Lectin mannose binding 1 5. DNA fragmentation factor 45 6. Tubulin specific chaperone C 7. Arylhydrocarbon-interacting receptor protein-like 1 8. Tubulin specific chaperone D 9. Unactive progesterone receptor 23KD (Yeah, That is its nameit is not printing mistake) Explanation A. Protein synthesis is a big process that involves a whole lot of enzymes. B. Protein degradation is by the digestive enzymes. C. Protein denaturation is usually by heat, organic solvents, mechanical mixing, strong acids or bases, detergents and ions of heavy metals such as lead and mercury. D. Protein folding is by chaperones. Comments Few more points about chaperones 1. Present in many species 2. Few are induced by conditions which cause unfolding of newly synthesised proteins 3. Bind predominantly to hydrophobic regions 4. Act as editing mechanisms or quality control 5. Most have ATPase activity 6. Found in: a. Cytosol b. Mitochondria c. Lumen of endoplasmic reticulum. Tips Read the enzymes involved in human as well as bacterial protein synthesis with relevance to pharmacology (Anti microbial and chemotherapeutic agent). 39. Which of the following is present intracellularly in muscle cells: A. Insulin. B. Corticosteroid. C. Epinephrine. D. Glucagon. Answer: B (Corticosteroid) Since all are present inside the cell, we read the question as for which of the following, the receptor is present inside the cell and then the answer will be. Ref: KDT 4th edn, 240,241; KDT 5th edn, 214; Ganong Quality: Spotter with a small mistake Status: Repeat QTDF: Ganong Discussion 1. Receptors for most of the hormones are in the cell membrane: Act by camp Adrenaline, glucagon, TSH, FSH, LH, PTH, ACTH, some hypothalamic releasing factors Act by IP3/DAG: Vasopressin Oxytocin Act by enzyme activation: Tyrosine LinaseInsulin Receptors of few are inside the cell: In cytoplasm: Steroids other than estrogen In nucleus: Estrogen (Refer Ganong Chapter 1 and the MCQs given at the back of the book in Chapter 1) Thyroid hormones T3, T4 Explanation A. Insulin acts by transmembrane enzyme activating receptor. B. Corticosteroid has intracellular receptors. C. Epinephrine acts by membrance receptor. D. Glucagon acts by membrane receptors. Comments After the exam, the answer for this question was not debated and it was found that most had attempted even though they knew that the question is wrong !!! Tips If you feel that there is a mistake in the question and ONLY one of the choices could be the right choice if the question was corrected then go for it. On the other hand, if more than one choice can turn out to be the correct answer for a correct question, you will be risking a negative mark. 40. All of the following statements are correct about potassium balance, except: A. Most of potassium is intracellular. B. Three quarter of the total body potassium is found in skeletal muscle. C. Intracellular potassium is released into extracellular space in response to severe injury. D. Acidosis leads to movement of potassium from extracellular to intracellular fluid compartment. Answer: D (Acidosis leads to movement of potassium from extracellular to intracellular fluid compartment) Ref: Ganong 20th edn, 30; Harrison 15th edn, 278 Quality: Reader Status: Repeat QTDF: Discussion Potassium is a major intracellular cation and is one of the determinants of resting membrane potential as well as the process of depolarisation and repolarisation. 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Nuclear magnetic resonance spectroscopy: It is a powerful and theoretically complex analytical tool technique used for determining the content and purity of a sample as well as its molecular structure. For example, NMR can quantitatively analyse mixtures containing known compounds. For unknown compounds, NMR can either be used to match against spectral libraries or to infer the basic structure directly. Once the basic structure is known, NMR can be used to determine molecular conformation in solution as well as studying physical properties at the molecular level such as conformational exchange, phase changes, solubility, and diffusion. In order to achieve the desired results, a variety of NMR techniques are available. D. X-ray diffraction: The term X-ray diffraction refers to the diffraction of X-rays by a crystal. The wavelengths of X-rays are comparable in size to the distances between atoms in most crystals, and the repeated pattern of the crystal lattice acts as a diffraction grating for X-rays. Comments The term X-ray powder diffraction refers in general to X-ray diffraction of a collimated monochromatic beam by a sample (a powder) containing a large number of tiny crystals having random orientations. The term X-ray crystallography refers to the use of X-ray diffraction to determine the structure of crystals or molecules. In general, the technique involves directing a beam of X-rays at a crystalline sample and recording the diffracted X-rays on a photographic plate. The X-ray diffraction pattern consists of a pattern of spots on the plate, and the crystal structure can be determined from the positions and intensities of the diffraction spots. Since X-rays are diffracted by the electrons in a molecule, if molecular crystals of a compound are used, the electron density distribution in the molecule can be determined. X-ray diffraction is currently one of the most important tools of solid-state physics and chemistry, the technique used for differentiation between crystalline and amorphous materials, determination of the structure of crystalline materials, determination of electron distributions within atoms and throughout unit cells of a crystal, determination of the orientation of single crystals, determination of the texture of polygrained materials, etc. In the biological sciences, x-ray diffraction has contributed greatly to our understanding of processes occurring in living cells, with the technique revealing the structures of proteins and nucleic acids and their constituent monomers, and also of drugs, hormones, and vitamins. Knowledge of 3-dimensional structures of various biomolecules has had a profound impact on the whole of biology. Tips Do you know that the following types of microscopes are used for identification purpose: 1. Stereoscopic microscope 2. Comparative microscope 3. Fluorescent microscope 4. Polarising microscope 5. Electron microscope 46. Porphobilinogen in urine produces pink colour with: A. Fouchets reagent. B. Benedicts reagent. C. Sodium nitropruside. D. Ehrlichs aldehyde reagent. Answer: D (Ehrlichs aldehyde reagent) Ref: J. Sengupta, Synopsis of Clinical Pathology & Microbiology, 5th, 82; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde, 477; Chaterjee and Sinde 2nd edn, 322 Chapter on Porphyrias Quality: Spotter Status: New QTDF: Chaterjee Discussion Watson and Schwartz test for porphobilinogen. Procedure 1. Mix equal volume of fresh urine and Ehrlich's reagent (0.7 gm of p-dimethyl amino benzaldehyde + 150 ml of conc HCl and 100 ml distilled water) 2. Allow to stand for 3 minutes 3. To this add saturated aqueous sodium acetate solution (2 volumes) and 4. Leave for 3 minutes 5. Add a few ml of chloroform 6. Shake thoroughly 7. Porphobilinogen forms a red aldehyde compound which (unlike that formed by urobilinogen) is insoluble in chloroform. Interpretation Red colour after second extraction with chloroform means a positive test for porphobilinogen and highly suggestive of acute intermittent porphyria. On the other hand, if the chloroform becomes pink, it is urobilinogen Explanation A. Fouchets reagent detects bile pigments. B. Benedicts reagentAll of you will know one Benedict's reagent, but there is another also !!! (See uric acid in the Table below) C. Sodium nitropruside is used along with ammonium sulphate for detection of ketones in rohtera test. D. Ehrlichs aldehyde reagent. Is used as above. Comments The table below is incomplete and should not be used for studying. A complete table is beyond the scope of this book, but may be found at www.rxpg.com, www.rxpgonline.com and www.targetpg.com. This is for refreshing your memory. Only the reagents are dealt with and the procedures are not given for want of space Few points about Urine Analysis Substance Name of Reagent Colour Detected the Test Chloride Fantus AgNO3 White Sulphate Acetic Acid, BaCl2 White Ethereal Acetic Acid, BaCl2 ,HCl Turbidity sulphates Indiacan Jaffe Hypochlorite (K chlorate Indigo blue and conc HCl) extract with Chloroform Oberneyers Ferric chloride + HCl Indigo blue extract with chloroform Calcium Sulkowich Oxalic acid, ammonium Fine white cloud or oxalate and glacial heavy milky white acetic acid Urea Hypobromite Sodium hypobromite Effervescence Specific Phenol red, 1% Faint yellow red Urease Acetic acid or 2% Na2CO3 Specific Above test + 1 ml of Intense pink colour Urease Urease + shake and wait for 10 minutes Doremus Ureometer Uric acid Benedicts Benedicts uric acid Blue colour Uric acid test Reagent + Na2CO3 Murexide conc HNO3 + Heat Reddish deposit Test Above test+ dip a rod Reddish violet in dil NH3 and draw it through the deposit Deposit + NaOH Bluish violet Protein Heat test Test tube heated Precipitate Sulpho- 25 % sulphosalicylic White precipitate salicylic acid Acid test Nitirc acid Conc HNO3 White ring test Contd... Contd... 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Structural haemoglobinopathies: occur when mutations alter the amino acid sequence of a globin chain, altering the physiologic properties of the variant haemoglobins and producing the characteristic clinical abnormalities. The variant haemoglobins relevant to this chapter: polymerise abnormally, as in sickle cell anaemia, or exhibit altered solubility or oxygen-binding affinity. 2. Thalassaemia syndromes arise from mutations that: impair production or translation of globin mRNA: leading to deficient globin chain biosynthesis. Clinical abnormalities are attributable: to the inadequate supply of hemoglobin and the imbalances in the production of individual globin chains, leading to premature destruction of erythroblasts and red cells. 3. Thalassaemic haemoglobin variants combine features of thalassaemia (e.g., abnormal globin biosynthesis) and of structural haemoglobinopathies (e.g., an abnormal amino acid sequence). 4. Hereditary persistence of foetal haemoglobin (HPFH) is characterised by: synthesis of high levels of foetal haemoglobin in adult life. 5. Acquired haemoglobinopathies include modifications of the haemoglobin molecu le by toxins (e.g., acquired methaemoglobinemia), and abnormal haemoglobin synthesis (e.g., high levels of HbF production in preleukaemia and a-thalassaemia in myeloproliferative disorders). Few details about HbS Haematology Normal in the heterozygote; haemolytic anaemia of varying degrees in homozygotes; painful vaso-occlusive episodes; leg ulcers; jaundice, stroke, congestive heart failure; meningitis; lowered resistance to salmonella and pneumococcus organisms Lectrophoresis HbS and HbA can readily be separated at both alkaline and acidic pH Chromatography HbS and HbA separate by both cation and anion exchange chromatography Structure Tryptic digestion; fingerprinting; cation studies exchange chromatography; amino acid analysis; sequencing; HPLC DNA Analyses A GAG->GTG mutation at codon 6 functions Normal studies Stability Normal; in the deoxy form HbS precipitates, particularly at higher salt concentrations Occurrence Heterozygotes and homozygotes found in many ethnic groups but predominantly in the Black race and in some Indian tribes Other HbS is the most studied Hb variant Information Explanation A. An abnormality in porphyrin part of haemoglobin will lead to porphyrias. B. Replacement of glutamate by valine in B-chain of HbA is the cause of HbS. C. A nonsense mutation in the B-chain of HbA will lead to chain termination. D. Substitution of valine by glutamate in the alpha chain of HbA will lead Hb Pavie in which valine in position 135 is replaced by glutamate. And the following table gives the haemoglobin variants caused by the substitution of valine by glutamate in the beta chain of HbA Hb Position Original AA Replaced Original Mutation AA Sequence Trollhttan 20(B2) Val Glu GTG GAG Cagliari 60(E4) Val Glu GTG GAG M-Milwaukee-I 67(E11) Val Glu GTG GAG Mainz 98(FG5) Val Glu GTG GAG New York 113(G15) Val Glu GTG GAG Hofu 126(H4) Val Glu GTG GAG North Shore 134(H12) Val Glu GTG GAG Comments Since most of the details for this are already in all the books, let us see few facts which are likely to be asked in future exams. The types and number of haemoglobin variants are: Type Number Alpha chain variants 199 Beta chain variants 335 Gamma chain variants (G-gamma = 38; A-gamma = 20; unknown = 3; special = 7) 68 Delta chain variants 28 Variants with two amino acid replacements (alpha = 1; beta = 17) 18 Variants with hybrid chains 10 Variants with elongated chains (at the C-terminus = 9; at the N-terminus = 4) 13 Variants with deletions (15); with insertions (4); with deletions and insertions (3) 22 Total 693 Few haemoglobins where the substitution occurs in position 6 in beta chain: Hb Position Original Replaced Original Mutation AA AA Sequence S 6(A3) Glu Val GAG GTG C 6(A3) Glu Lys GAG AAG G-Makassar 6(A3) Glu Ala GAG GCG Machida 6(A3) Glu Gln GAG CAG One more alpha chain disorder which is: Ann Arbor 80(F1) Leu->Arg (CTG > CGG) Tips Refer Table 106 1 in Harrison 15th Edition and the relevant chapters in Nelson. For a complete list visit www.rxpgonline.com, www.targetpg.com 31. Decreased glycolytic activity impairs oxygen transport by haemoglobin due to: A. Reduced energy production. B. Decreased production of 2,3-bisphosphoglycerate. C. Reduced synthesis of haemoglobin. D. Low levels of oxygen. Answer: B (Decreased production of 2,3-bisphosphoglycerate) Ref: Harper 25th,70, 195; Lippincott 2nd edn, 31,92 Figure 7.8; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde, 258 Quality: Reader Status: Thinker QTDF: Harper Discussion 2, 3 DPG (2, 3-biphosphoglycerate) is produced by the alternate pathway and when the glycolytic activity decreases, the amount of 2, 3 DPG decreases and we all know that 2, 3-DPG releases oxygen from Hb to the tissues. Thus when there is low amount of 2, 3-DPG, there is shift to left, that is oxygen is bound tightly to the Hb and as a result there is decreased availability of oxygen to the tissues. Explanation A. Oxygen delivery is not related to energy production. B. Decreased production of 2,3-bisphosphoglycerate leads to shift to left. C. Decreased glycolytic activity does not reduce synthesis of haemoglobin all of a sudden. D. Low levels of oxygen leads to decreased glycolytic activity (of course) but not the opposite as given in this choice. 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(xKddh(xKdddH(xKddd(xKddd#(xKdddh(xKddd (xKddd(xKddd(xKdddh(xKddd(xKddd(xKddd (xKddd%h(xKddd ,(xKdd(X(xKddd:X(xKdddJX(xKdddCX(xKddd ,(xKddfh(xKddd ,@ l 8,@ l H,@ l ,@ l ,@ l M,@ l <,@ l G,@ l }hthXhUh[hhqh ,@ l ,@ l ,@ l I,@ l ,@ l hh XX%XX,@ l 8,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l XMXKX,=#C</1/ 8G @{.@@.-@c-00%%((@    @     @@U  @ @ "      u @  @{ @?  2@  @@ &%   @6 6     @    5 @ +  @    @   @@8 :      @W , @ @; ;  6@@B@XY'.@A@(`?)6bT@*@-0E@F\@,In acidosis, the pump mechanism is altered and the intracellular potassium wanders out of its home into the ECF During tissue injury, due to the loss of cell membrane integrity, hyperkalaemia occurs Hypokalaemia (Increased potassium inside cells): a. Insulin b. Aldosterone c. Alkalosis d. Beta adrenergic drugs (Salbutamol) Hyperkalaemia (Decreased potassium outside cells): a. Diabetes b. Addison's c. Acidosis d. Beta blockersPropanolol e. Cell injury f. Exercises Explanation A. Most of potassium is intracellular. B. Three quarter of the total body potassium is found in skeletal muscle. C. Intracellular potassium is released into extracellular space in response to severe injury remember that rhabdomyolysis leads to hyperkalaemia. D. Acidosis leads to movement of potassium from intracellular to extracellular fluid compartment. Comments There will be no question paper without a question on sodium, potassium, calcium, acidosis, alkalosis (Acid and Electrolyte Balance), etc. In any question paper, you can expect 5% of questions on this. Tips Prepare well. Read the relevant chapters in all books, For certain topics, it is enough if you read any one of the book, but for this you have to read all the concerned books. 41. Causes of metabolic alkalosis include all the following, except: A. Mineralocorticoid deficiency. B. Bartters syndrome. C. Thiazide diuretic therapy. D. Recurrent vomiting. Answer: A (Mineralocorticoid deficiency) Ref: Harrison 15th edn, Chapter 50, Table 50.4 and 288 Quality: Reader Status: Repeat QTDF: Harrison Discussion Metabolic alkalosis occurs as a result of net gain of [HCO3] or loss of nonvolatile acid (usually HCl by vomiting) from the extracellular fluid. Since it is unusual for alkali to be added to the body, the disorder involves a generative stage, in which the loss of acid usually causes alkalosis, and a maintenance stage, in which the kidneys fail to compensate by excreting HCO3 because of volume contraction, a low GFR, or depletion of Cl or K. Under normal circumstances, the kidneys have an impressive capacity to excrete HCO3. Continuation of metabolic alkalosis represents a failure of the kidneys to eliminate HCO3 in the usual manner. For HCO3 to be added to the extracellular fluid, it must be administered exogenously or synthesised endogenously, in part or entirely by the kidneys. The kidneys will retain, rather than excrete, the excess alkali and maintain the alkalosis if: 1. volume deficiency, chloride deficiency, and K+ deficiency exist in combination with a reduced GFR, which augments distal tubule H+ secretion; or 2. hypokalaemia exists because of autonomous hyperaldosteronism. In the first example, alkalosis is corrected by administration of NaCl and KCl, while in the latter it is necessary to repair the alkalosis by pharmacologic or surgical intervention, not with saline administration. Explanation A. In patients with decreased mineralocorticoid production or action, disorders of aldosterone biosynthesis or action are associated with high renin levels, salt wasting, and hyperkalaemia. The aldosterone levels may be low or elevated. In patients with a deficiency in aldosterone biosynthesis, the transformation of corticosterone into aldosterone is impaired, owing to a mutation in the aldosterone synthase (CYP11B2) gene. These patients have low to absent aldosterone secretion, elevated plasma renin levels, and elevated levels of the intermediates of aldosterone biosynthesis (corticosterone and 18-hydroxycorticosterone). Pseudohypoaldosteronism type I (PHA-I) is an autosomal recessive disorder that is seen in the neonatal period and is characterised by salt wasting, hypotension, hyperkalemia, and high renin and aldosterone levels. In contrast to the gain-of-function mutations in the epithelial sodium channel (ENaC) in Liddles syndrome, mutations in PHA-I result in loss of ENaC function. 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Its importance derives from the fact that glucose uptake by the liver is an essential physiological process, crucial for glucose homeostasis. And it can phosphorylate glucose only and cannot be involved in fructose metabolism. Comments This enzyme is also called NAD-dependent glyceraldehyde-3-phosphate dehydrogenase and triosephosphate dehydrogenase and it also acts very slowly on D-glyceraldehyde and some other aldehydes. Thiols can replace phosphate. Tips The reaction catalysed by glyceraldehyde-3-phosphate dehydrogenase catalyses is an reaction important in the generation of ATP. Make a note of all the reactions in various pathways in which high energy phosphates and NAD/NADP are involved. 33. B-oxidation of odd-chain fatty acids produces: A. Succinyl CoA. B. Propionyl CoA. C. Acetyl CoA. D. Malonyl CoA. Answer: B (Propionyl CoA) Ref: Harper 25th, 240; Lippincott 2nd edn, 183, 184 Figure 17.17; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 381; Chaterjee 2nd edn, 542. MCQ given at the end of the chapter 20 Quality: SpotterFirst MBBS Status: Repeat QTDF: ChaterjeeMCQ given at the end of the chapter Discussion Beta oxidarion of even chain fatty acids produce acetly CoA and that of odd chain fatty acids producs acetyl CoA and one propionyl CoA. Explanation A. Succinyl CoA is not produced. B. One molecule of propionyl CoA is produced. C. Acetyl CoA also is produced but it is produced in odd as well as even chain chain fatty acids. D. Malonyl CoA is not produced. Comments A question that is repeated for a long period of time. Tips Go through the MCQs given at the end of each chapters in Chatterjee. Questions are being asked from that repeatedly. 34. The main enzyme responsible for activation of xenobiotics is: A. Cytochrome P-450 B. Glutathione S-transferase. C. NADPH cytochrome P-450-reductase. D. Glucuronyl transferase. Answer: A (Cytochrome P-450) Ref: Harper 24th edn, Chapter 61 Metabolism of Xenobiotics, 750; Harper 25th edn, 781; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 487 Quality: Thinker !!! Status: New QTDF: Harper Discussion A xenobiotic (Gk xenos stranger) is a compound that is foreign to the body, the principal classes of xenobiotics of medical relevance are drugs, chemical carcinogens, and various compounds that have found their way into our environment. It is convenient to consider the metabolim of xenobiotics in two phases. In phase 1, the major reaction involved is hydroxylation, catalysed by members of a class of enzymes referred to as mono-oxygenases or cytochrome P-450s. Hydroxylation may terminate the action of a drug, though this is not always the case. In addition to hydroxylation, these enzymes catalyse an astonishingly a wide range of reactions, including those involving deamination, dehalogenation, desulphuration, epoxidation, peroxygenation, and reduction. Reactions involving hydolysis (e.g., catalysed by esterases) and certain other non-P450-catalysed reactions also occur in phase 1. In phase 2, the hydroxylated or other compounds produced in phase 1 are converted by specific enzymes to various polar metabolites by conjugation with glucuronic acid, sulphate, acetate, glutathione or certain amino acids or methylation. Oxidation by mono-oxygenases proceed as the following reaction, catalysed by cytochrome P-450 (and in the process of which cytochrome P-450 turn into oxidised P-450. Reduced cytochrome P-450 + RH + O2 R-OH + H2O + Oxidised Cytochrome P-450 And here in this reaction, which is the reaction responsible for activation of xenobiotics (asked in the question) cytochrome P-450 is involved and then there is another reaction in which NADPH, not NADH is involved. It is in the reaction mechanism of cytochrome P-450. The enzyme that uses NADPH to yield the reduced cytochrome P-450, shown in the left hand side of the above equation, is called NADPH - cytochrome P-450 reductase. Electron, are transferred from NADPH to NADPH-cytochrome P-450 reductase and then to cytochrome P-450. 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In antenatal and classic Bartters syndrome, impaired Cl reabsorption in the thick ascending limb of the loop of Henle is the underlying defect. Inadequate Cl reabsorption causes volume depletion and activates the renin-angiotensin system. Distal delivery of NaCl and water are high in the presence of high aldosterone, promoting secretion of K+ and H+ ions. Prostaglandin overproduction is mediated by volume depletion, hypokalaemia, and high angiotensin II and kallikrein levels. Increased prostaglandin production contributes to the severity of disease by inducing resistance to the pressor effects of angiotensin II and reducing reabsorption in the thick ascending limb of the loop of Henle. Mutations in the bumetanide-sensitive Na:K:2Cl channel, the apical ATP-regulated K+ channel, and the basolateral Cl channel have been described in classic and antenatal Bartters. C. Thiazide diuretics increase urinary excretion of sodium and water by inhibiting sodium reabsorption in the early distal tubules. They increase the rate of delivery of tubular fluid and electrolytes to the distal sites of hydrogen and potassium ion secretion, while plasma volume contraction increases aldosterone production. The increased delivery and increase in aldosterone levels promote sodium reabsorption at the distal tubules, thus increasing the loss of potassium and hydrogen ions. And this leads to alkalosis. D. Recurrent vomiting leads to loss of acid and that leads to alkalosis. Comments Metabolic acidosis leads to hyperkalaemia as a result of cellular shifts in which H+ is exchanged for K+ or Na+. For each decrease in blood pH of 0.10, the plasma [K+] should rise by 0.6 mmol/L. This relationship is not invariable. Diabetic ketoacidosis, lactic acidosis, diarrhoea, and renal tubular acidosis (RTA) are often associated with potassium depletion because of urinary K+ wasting. Tips Read about RTA (Renal Tubular Acidosis) in Harrison. 42. Normal anion gap metabolic acidosis is caused by: A. Cholera. B. Starvation. C. Ethylene glycol poisoning. D. Lactic acidosis. Answer: A (Cholera) Ref: Harrison 15th edn, Chapter 50 and, 285 Quality: Reader Status: Repeat QTDF: Harrison Discussion There are four principal causes of a high-anion gap acidosis: 1. Lactic acidosis: a. Anaerobic metabolism predominatingshock/Cardiopulmonary arrest b. Severe anaemia, c. Poisoning with CO or cyanide, 2. Ketoacidosis: a. Diabetes b. Alcoholics c. Starvation 3. Ingested toxins: a. Ethylene glycol b. Salicylates c. Methanol 4. Acute and chronic renal failure: Explanation A. In cholera the anion gap is normal. B. Starvation is a cause of increased anion gap acidosis C. Ethylene glycol poisoning is a cause of increased anion gap acidosis D. Lactic acidosis is a cause of increased anion gap acidosis. Comments All evaluations of acid-base disorders should include a simple calculation of the anion gap (AG); It represents those unmeasured anions in plasma (normally 10 to 12 mmol/L) It is calculated as follows: AG = Na+ - (Cl + HCO3). The unmeasured anions include anionic proteins, phosphate, sulphate, and organic anions. When acid anions, such as acetoacetate and lactate, accumulate in extracellular fluid, the AG increases, causing a high-AG acidosis. An increase in the AG is most often due to an increase in unmeasured anions and less commonly is due to a decrease in unmeasured cations (calcium, magnesium, potassium). In addition, the AG may increase with an increase in anionic albumin, either because of increased albumin concentration or alkalosis, which alters albumin charge. A decrease in the AG can be due to: An increase in unmeasured cations; The addition to the blood of abnormal cations, such as lithium (lithium intoxication) or cationic immunoglobulins (plasma cell dyscrasias); A reduction in the major plasma anion albumin concentration (nephrotic syndrome); A decrease in the effective anionic charge on albumin by acidosis; or Hyperviscosity and severe hyperlipidaemia, which can lead to an underestimation of sodium and chloride concentrations. Tips Other conditions with normal anion gap are 1. Diarrhoea 2. Fistula 3. Ureterosigmoidostomy 4. Renal tubular acidosis 5. Ingestion of ammonium chloride 6. Mineralocorticoid deficiency. 43. A 50 kg. man with severe metabolic acidosis has the following parameters:pH 7.05. pCO2 12 mm Hg., pO2 108 mmHg. HCO3 5 mEq/L. base excess-30 mEq/L. The approximate quantity of sodium bicarbonate that he should receive in half hour is: A. 250 mEq. B. 350 mEq. C. 500 mEq. D. 750 mEq. Answer: A (250 mEq) Ref: Harrison 15th edn, Chapter Quality: Reader Status: Repeat QTDF: Harrison Discussion Bicarbonate replacement Not usually recommended unless pH < 7.2 or HCO3- deficit is greater than 5 mEq/L. 8.4% NaHCO3 = 1 mEq/ml 1 gm of baking soda = 12 mEq of NaHCO3 Formula for bicarbonate replacement 0.3 (BW in Kg) (HCO3- deficit) = mEq of NaHCO3 needed 0.3 assumes 30% of BW is extracellular fluid. If the figure of 0.3 is used, bicarbonate replacement can be given fairly rapidly (30 minutes to 1 hour). If figure of .5 is used bicarbonate replacement should be given over 12-24 hour period and half of it should be given in the first half hour If you have a mixed respiratory/metabolic acidosis, important to address the respiratory acidosis first, before treating the metabolic acidosis Explanation In our case replacement = 0.5 Body Weight (Desired HCO3 Measured HCO3) Replacement in hour = 0.25 Body Weight (Desired HCO3 Measured HCO3) = 0.25 50 (25-5) = 250 mEq Comments The base excess is a calculated figure which provides an estimate of the metabolic component of the acid-base balance. The base excess is defined as the amount of H+ ions that would be required to return the pH of the blood to 7.35 if the PCO2 were adjusted to normal. Because the base excess is a calculated (not a measured) value, it may be inaccurate and misleading. Despite these problems, it is important to understand the concept. 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The molecular weight of all isoenzymes is ca. 140 kDa. L(+)-lactate dehydrogenase is specific for L(+)-lactate and does not react with D()-lactate. LDH is used in coupled enzyme assays, for example, in the determination of ATPase, myokinase and pyruvatekinase. It may also be used in the determination of lactate (5), pyruvate (6) and various other metabolites. D. Glucokinase [Hexokinase D (EC 2.7.1.1)] is the characteristic isoenzyme of hexokinase in the liver, and it is responsible for glucose phosphorylation in hepatocytes. Its importance derives from the fact that glucose uptake by the liver is an essential physiological process, crucial for glucose homeostasis. And it can phosphorylate glucose only and cannot be involved in fructose metabolism. Comments This enzyme is also called NAD-dependent glyceraldehyde-3-phosphate dehydrogenase and triosephosphate dehydrogenase and it also acts very slowly on D-glyceraldehyde and some other aldehydes. Thiols can replace phosphate. Tips The reaction catalysed by glyceraldehyde-3-phosphate dehydrogenase catalyses is an reaction important in the generation of ATP. Make a note of all the reactions in various pathways in which high energy phosphates and NAD/NADP are involved. 33. B-oxidation of odd-chain fatty acids produces: A. Succinyl CoA. B. Propionyl CoA. C. Acetyl CoA. D. Malonyl CoA. Answer: B (Propionyl CoA) Ref: Harper 25th, 240; Lippincott 2nd edn, 183, 184 Figure 17.17; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 381; Chaterjee 2nd edn, 542. MCQ given at the end of the chapter 20 Quality: SpotterFirst MBBS Status: Repeat QTDF: ChaterjeeMCQ given at the end of the chapter Discussion Beta oxidarion of even chain fatty acids produce acetly CoA and that of odd chain fatty acids producs acetyl CoA and one propionyl CoA. Explanation A. Succinyl CoA is not produced. B. One molecule of propionyl CoA is produced. C. Acetyl CoA also is produced but it is produced in odd as well as even chain chain fatty acids. D. Malonyl CoA is not produced. Comments A question that is repeated for a long period of time. Tips Go through the MCQs given at the end of each chapters in Chatterjee. Questions are being asked from that repeatedly. 34. The main enzyme responsible for activation of xenobiotics is: A. Cytochrome P-450 B. Glutathione S-transferase. C. NADPH cytochrome P-450-reductase. D. Glucuronyl transferase. Answer: A (Cytochrome P-450) Ref: Harper 24th edn, Chapter 61 Metabolism of Xenobiotics, 750; Harper 25th edn, 781; Textbook of Medical Biochemistry 4th edn, Chaterjee and Shinde 487 Quality: Thinker !!! Status: New QTDF: Harper Discussion A xenobiotic (Gk xenos stranger) is a compound that is foreign to the body, the principal classes of xenobiotics of medical relevance are drugs, chemical carcinogens, and various compounds that have found their way into our environment. It is convenient to consider the metabolim of xenobiotics in two phases. In phase 1, the major reaction involved is hydroxylation, catalysed by members of a class of enzymes referred to as mono-oxygenases or cytochrome P-450s. Hydroxylation may terminate the action of a drug, though this is not always the case. In addition to hydroxylation, these enzymes catalyse an astonishingly a wide range of reactions, including those involving deamination, dehalogenation, desulphuration, epoxidation, peroxygenation, and reduction. Reactions involving hydolysis (e.g., catalysed by esterases) and certain other non-P450-catalysed reactions also occur in phase 1. In phase 2, the hydroxylated or other compounds produced in phase 1 are converted by specific enzymes to various polar metabolites by conjugation with glucuronic acid, sulphate, acetate, glutathione or certain amino acids or methylation. Oxidation by mono-oxygenases proceed as the following reaction, catalysed by cytochrome P-450 (and in the process of which cytochrome P-450 turn into oxidised P-450. Reduced cytochrome P-450 + RH + O2 R-OH + H2O + Oxidised Cytochrome P-450 And here in this reaction, which is the reaction responsible for activation of xenobiotics (asked in the question) cytochrome P-450 is involved and then there is another reaction in which NADPH, not NADH is involved. It is in the reaction mechanism of cytochrome P-450. The enzyme that uses NADPH to yield the reduced cytochrome P-450, shown in the left hand side of the above equation, is called NADPH - cytochrome P-450 reductase. Electron are transferred from NADPH to NADPH-cytochrome P-450 reductase and then to cytochrome P-450. Now let us see what NADPH-cytochrome P-450 reductase does NADPH + Oxidised cytochrome P-450 ,@ l 8,@ l H,@ l ,@ l ,@ l M,@ l <,@ l G,@ l }hthXhUh[hhqh ,@ l ,@ l ,@ l I,@ l ,@ l hh XX%XX,@ l 8,@ l ,@ l ,@ l ,@ l ,@ l ,@ l ,@ l XMXKXPp reduced cytochrome P-450 + NADP This reaction generates reduced P-450 for the first reaction, i.e. it activates cytochrome P-450 which is an inherent part of our body. Explanation A. Cytochrome P-450 : the chief enzyme involved in activation of Strangers is mono-oxygenase or cytochrome P-450. NADPH-cytochrome P-450 reductase is involved in the reactivation of cytochrome P-450 which is not a stranger. B. Glutathione S-transferase is an enzyme involved in activation of xenobiotics, Harper has clearly stated that of all Phase 1 reactions it is hydroxylation (mono-oxygenation - Monooxygenatiuon means adding one oxygen. When that oxygen is added to a R-H group, we get R-OH and because of that -OH we call it as hydroxylation) which is important. Hence, there is no doubt regarding this enzyme is not the main enzyme. C. NADPH cytochrome P-450-reductase.is involved, but not in activation of xenobiotics: Xenobiotics is (Gk xenos stranger) is a compound that is foreign to the body, the principal classes of xenobiotics of medical relevance are drugs, chemical carcinogens. Obviously, P-450 is not a xenobiotic !!!! Then how come an enzyme (which activates another enzyme, which is in our body) be a main enzyme in the activation of xenobiotics !!!! Again the question is clear.. Activation of xenobiotics. Atleast if the question is concerned with xenobiotics,.. there can be a little confusion. D. Glucuronyl transferase is an enzyme involved in activation of xenobiotics, but it is not the main enzyme. Comments The confusa in this question is due to the line NADPH, not NADH is involved in the reaction mechanism of cytochrome P-450 in Harper. But as explained, NADPH and NADH is used, but not used in the activation of xenobiotics. Tips Xenobiotics is a high yield topic in both biochemistry and pharmacology. 35. The protein rich in basic amino acids, which functions in the packaging of DNA in chromosomes, is: A. Histone. B. Collagen. C. Hyaluronic acid-binding protein. D. Fibrinogen. Answer: A (Histone) Ref: Harper 25th edn, 412 Quality: Spotter Status: Repeat QTDF: Harper Discussion Histones are proteins found in the nuclei of all eukaryotic cells where they are complexed to DNA in chromatin and chromosomes. They are of relatively low molecular weight and are basic, having a very high arginine/lysine content. They are highly conserved and can be grouped into five major classes. Two copies of H2A, H2B, H3 and H4 bind to about 200 base pairs of DNA to form the repeating structure of chromatin, the nucleosome, with H1 binding to the linker sequence. They may act as non-specific repressors of gene transcription. Explanation A. Histones. B. Collagen is the protein substance of the white fibres (collagenous fibres) of skin, tendon, bone, cartilage and all other connective tissue, composed of molecules of tropocollagen, it is converted into gelatin by boiling. The word has originated from the Greek word Kolla = glue and gennan = to produce are the structural mole. C. Hyaluronic acid-binding protein occurs in pharyngeal epithelial cells. The capsular polysaccharide of Group A Streptococcus plays a role in colonisation of the pharynx by binding to CD44, a hyaluronic acid-binding protein expressed on human pharyngeal epithelial cells. Though this is not related to this question, I have given this because this may be asked in future. Hyaluronic acid, binding protein 2 is a serine protease. D. Fibrinogen is soluble plasma protein (340 kD, 46 nm long), composed of 6 peptide chains (2 each of A_, B_ and _) and present at about 2-3 mg/ml. Comments Questions from genetics are asked directly from Harper and are often taken from the blue headings given before each paragraph. Tips If you dont have time to read Harper thoroughly, read at least the blue headings given at the top of each paragraph. 36. Which of the following is not a post-transcriptional modification of RNA? A. Splicing. B. 5' capping. C. 3' polyadenylation. D. Glycosylation. Answer: D (Glycosylation) Ref: Harper 25th edn, 448; Lippincott 2nd edn, 385,386 Quality: Reader Status: Repeat QTDF: Lippincott Fig. 31.14, 31.15. Discussion Transcription, as we all know, is a step in the synthesis of mRNA from DNA. First heteronuclear RNA (hnRNA) is synthesised (which in nothing but a reverse or complimentary copy of the DNA from which it was synthesised) the sequence of which is exactly same as the other strand of DNA (except for the Uracil), and it undergoes few changes, (which are called as post transcriptional modifications) to become an active mRNA. The changes are: 1. Additions 5 cappingAddition of 7 methyl guanosine capping to Initiate translation Protection of 5 end from attack of 5-3 exonuclease 3 cappingAddition of poly a tail or 3 polyadenylation 2. SplicingRemoval of introns and Merging the exons Explanation A. 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