Biochemistry

43 - Ninhydrin test or Ninhydrin reagent
Ninhydrin (2,2-Dihydroxyindane-1,3-dione) is a chemical used to detect ammonia or

primary and secondary amines. When reacting with these free amines, a deep blue or purple
color known as Ruhemann's purple is produced. Ninhydrin is most commonly used to
detect fingerprints, as the terminal amines or lysine residues in peptides and proteins
sloughed off in fingerprints react with ninhydrin.
Ninhydrin can also be used to monitor deprotection in solid phase peptide synthesis (Kaiser
Test). The chain is linked via its C-terminus to the solid support, with the N-terminus
extending off it. When that nitrogen is deprotected, a ninhydrin test yields blue. Amino-acid
residues are attached with their N-terminus protected, so if the next residue has been
successfully coupled onto the chain, the test gives a colorless or yellow result.
Ninhydrin is also used in amino acid analysis of proteins: Most of the amino acids are
hydrolyzed and reacted with ninhydrin except proline; Also, certain amino acid chains are
degraded. Therefore, separate analysis is required for identifying such amino acids that
either react differently or don't react at all with ninhydrin. The rest of the amino acids are
then quantified colorimetrically after separation by chromatography.
A solution suspected of containing the ammonium ion can be tested by ninhydrin by dotting
it onto a solid support (such as silica gel); treatment with ninhydrin should result in a
dramatic purple color if the solution contains this species. In the analysis of a chemical
reaction by thin layer chromatography (TLC), the reagent can also be used. It will detect, on
the TLC plate, virtually all amines, carbamates and also, after vigorous heating, amides.
When ninhydrin reacts with amino acids, the reaction also releases CO2. The carbon in this
CO2 originates from the carboxyl carbon of the amino acid. This reaction has been used to
release the carboxyl carbons of bone collagen from ancient bones for stable isotope analysis
in order to help reconstruct the palaeodiet of cave bears.
A ninhydrin solution is commonly used by forensic investigators in the analysis of latent

fingerprints on porous surfaces such as paper. Amino acid containing fingermarks, formed
by minute sweat secretions which gather on the finger's unique ridges, are treated with the
ninhydrin solution which turns the amino acid finger ridge patterns purple and therefore
visible.
The carbon atom of a carbonyl bears a partial positive charge enhanced by neighboring
electron withdrawing groups like carbonyl itself. So the central carbon of a 1,2,3-tricarbonyl
compound is much more electrophilic than one in a simple ketone. Thus indane-1,2,3-trione
reacts readily with nucleophiles, including water. Whereas for most carbonyl compounds, a
carbonyl form is more stable than a product of water addition (hydrate), ninhydrin forms a
stable hydrate of the central carbon because of the destabilizing effect of the adjacent
carbonyl groups.
Note that in order to generate the ninhydrin chromophore, the amine is condensed with a
molecule of ninhydrin to give a Schiff base. Thus only ammonia and primary amines can
proceed past this step. At this step, there must also be an alpha proton (H* in the diagram)
for Schiff base transfer, so an amine adjacent to a tertiary carbon cannot be detected by the
ninhydrin test. The reaction of ninhydrin with secondary amines gives an iminium salt,
which is also coloured, and this is generally yellow-orange in color.
Posted by doctor at 6:55 AM 0 comments Links to this post

Labels: biochemistry lab tests, biuret test, lab tests to detect proteins, millon's reagent, millon's
test, ninhydrin reagent, ninhydrin test
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42 - Biuret test or Biuret reagent
The biuret test is a chemical test used for detecting the presence of peptide bonds. In the
presence of peptides, a copper(II) ion forms a violet-colored complex in an alkaline solution.
Several variants on the test have been developed.
The Biuret reaction can be used to assay the concentration of proteins because peptide
bonds occur with the same frequency per amino acid in the peptide. The intensity of the
color, and hence the absorption at 540 nm, is directly proportional to the protein
concentration, according to the Beer-Lambert law.
In spite of its name, the reagent does not in fact contain biuret ((H2N-CO-)2NH). The test is
so named because it also gives a positive reaction to the peptide bonds in the biuret
molecule.
Procedure :
An aqueous sample is treated with an equal volume of 1% strong base (sodium or potassium
hydroxide most often) followed by a few drops of aqueous copper(II) sulfate. If the solution
turns purple, protein is present. 5–160 mg/mL can be determined.
Biuret reagent :
The biuret reagent is made of potassium hydroxide (KOH) and hydrated copper (II) sulfate,
together with potassium sodium tartrate. The reagent turns from blue to violet in the
presence of proteins, blue to pink when combined with short-chain polypeptides.
Not all biuret tests require the biuret reagent. The reagent is commonly used in a biuret
protein assay, a colorimetric assay used to determine protein concentration—such as UV-
VIS at wavelength 540 nm (to detect the Cu2+ ion).
Increasing the sensitivity of the biuret test :
Cu+ is a strong reducing agent which can react for example with Mo(VI) in Folin-Ciocalteu's
reagent to form molybdenum blue. In this way, proteins can be detected in concentrations
between 0.005 and 2 mg/mL. Molybdenum blue in turn can bind certain organic dyes
(malachite green, Auramin O), resulting in further amplification of the of the signal.
Cu+ forms a deep purple complex with bicinchoninic acid (BCA), which allows proteins in
the range of 0.0005 to 2 mg/mL to be determined. This assay is often referred to as "Pierce
assay" after the manufacturer of a reagent kit.

Labels: beer-lambert law, biuret reagent, biuret test, detecting peptide bonds, lab tests to detect
proteins
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41 - Millon's test or Millon's reagent
Millon's reagent is an analytical reagent used to detect the presence of soluble proteins.
A few drops of the reagent are added to the test solution, which is then heated gently.
A reddish-brown coloration or precipitate indicates the presence of tyrosine residues which

occur in nearly all proteins.
Millon's test is not specific for proteins (it actually detects phenolic compounds), and so
must be confirmed by other tests for proteins such as the biuret test and the ninhydrin
reaction.
The reagent is made by dissolving metallic mercury in nitric acid and diluting with water.
The test was developed by the French chemist Auguste Millon (1812–67).

Labels: biuret test, detecting peptide bonds, detecting proteins, millon's reagent,millon's
test, ninhydrin reagent, ninhydrin test
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MONDAY, FEBRUARY 22, 2010
40 - Tetrahydrobiopterin (THB)
*Tetrahydrobiopterin (THB, BH4; Kuvan) or sapropterin, is a naturally occurring nutrient
and essential cofactor of the three aromatic amino acid hydroxylase enzymes, used in the
biosynthesis of the neurotransmitters serotonin (5-hydroxytryptamine (5-HT)), melatonin,
dopamine, norepinephrine (noradrenaline), epinephrine (adrenaline), and nitric oxide
(NO).
*THB was discovered to play a role as an enzymatic cofactor by Seymour Kaufman. The first
enzyme found to use THB is phenylalanine hydroxylase (PAH).
*THB is biosynthesized from guanosine triphosphate (GTP) by three chemical reactions,

those of which are mediated by the enzymes (GTP cyclohydrolase I (GTPCH), 6-
pyruvoyltetrahydropterin synthase (PTPS), and sepiapterin reductase (SPR).
*THB has the following responsibilities as a cofactor:
- Tryptophan hydroxylase (TPH) for the conversion of L-tryptophan (TRP) to 5-

hydroxytryptophan (5-HTP)
- Phenylalanine hydroxylase (PAH) for conversion of L-phenylalanine (PHE) to L-tyrosine
(TYR)
- Tyrosine hydroxylase (TH) for the conversion of L-tyrosine to L-DOPA (DOPA)
- Nitric oxide synthase (NOS) for conversion of a guanidino nitrogen of L-arginine (L-Arg)
to nitric oxide (NO)
- Glyceryl ether monooxygenase (GEMO) for the conversion of 1-alkyl-sn-glycerol to 1-
hydroxyalkyl-sn-glycerol.
*A deficit in THB biosynthesis and/or regeneration results in phenylketonuria (PKU) from
excess L-phenylalanine concentrations or hyperphenylalaninemia (HPA), as well as
monoamine and nitric oxide neurotransmitter deficiency or chemical imbalance. The
chronic presence of PKU can result in severe brain damage, including symptoms of mental
retardation, microcephaly, speech impediments such as stuttering, slurring, and lisps,
seizures or convulsions, and behavioral abnormalities, among other effects.
*THB, developed by BioMarin under the brand name Kuvan and approved by the United
States (U.S.) Food and Drug Administration (FDA) on December 13, 2007, is a synthetic
preparation of the dihydrochloride salt of the substance, used in the treatment of PKU. HPA
due to THB deficiency-mediated PKU.
*According to researchers at the University of Illinois at Chicago, THB may be of benefit in

the management and treatment of intractable systolic heart failure based on research in
mice. This is especially significant given that THB has already been tested and determined
to be safe in human patients using it in the management of phenylketonuria, and also
because there are far fewer ways to manage systolic heart failure than for diastolic heart
failure.
Posted by doctor at 10:39 PM 0 comments Links to this post

Labels: aromatic amine hydroxylases, bh4 functions, kuvan, reactions where thb acts as a
cofactor, sapropterin, Tetrahydrobiopterin functions, thb functions
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39 - Type I and Type II Skeletal Muscle Fibers
*A Sprinter Uses Creatine Phosphate and Anaerobic Glycolysis to Make ATP, Whereas a
Marathon Runner Uses Oxidative Phosphorylation.
Labels: Difference between energy source for sprinter and Marathon runner,Type 1 and Type 2
skeletal muscle fibers differences, Types of muscle fibers
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THURSDAY, JANUARY 28, 2010
38 - Catecholamine synthesis
37 - Biochemical reactions of Folate coenzymes
Reaction Coenzyme Form Importance
of Folate
Involved and
Single Carbon
unit transferred
Formate activation THF; -CHO Generation of
group 10-formyl-THF
transferred.
Purine synthesis
Formation of glycinamide 5,10- Formation of

ribonucleotide MethyleneTHF;- purines needed
CHO group for DNA, RNA
transferred synthesis, but
10-Formyl reactions
Formylation of probably not
aminoimidazolecarboxamide- (CHO)THF;-
CHO group rate limiting
ribonucleotide (AICAR) transferred
Pyrimidine synthesis
Methylation of deoxyuridine 5,10- Rate limiting in

monophosphate (dUMP) to MethyleneTHF;- DNA synthesis
CH3 group
thymidine monophosphate transferred.
Oxidizes THF
(dTMP) to DHF
Some
breakdown
of folate at the
C-9–N-10 bond
Amino acid interconversion
Serine–glycine THF; =CH2 Entry of single

interconversion group carbon units
transferred into active pool
Homocysteine to methionine 5- Demethylation
Methyl(M)THF;- of 5-MTHF to
CH3 group THF; also
transferred requires
cobalamin,
flavine adenine
dinucleotide,
ATP, and
adenosyl
methionine
Forminoglutamic acid to THF; -HN-CH=
glutamic acid in histidine group
transferred
catabolism

Labels: biochemical reactions of folate coenzymes, folate associated reactions,folate biochemistry
mcqs, formation of glycinamide ribonucleotide, single carbon units transferred in folate reactions
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WEDNESDAY, SEPTEMBER 16, 2009
36 - Diseases caused by mutations in collagen genes

Diseases Caused by Mutations in Collagen Genes or by
Deficiencies in the Activities of Posttranslational Enzymes
Involved in the Biosynthesis of Collagen.
Gene or Enzyme Disease
COL1A1, Osteogenesis imperfecta, type 1 (MIM
COL1A2 166200)
Osteoporosis (MIM 166710)
Ehlers-Danlos syndrome type VII autosomal
dominant (130060)
COL2A1 Severe chondrodysplasias
Osteoarthritis (MIM 165720)
COL3A1 Ehlers-Danlos syndrome type IV (MIM
130050)
COL4A3–COL4A6 Alport syndrome (including both autosomal
and X-linked forms) (MIM 104200)
COL7A1 Epidermolysis bullosa, dystrophic (MIM
131750)
COL10A1 Schmid metaphysial chondrodysplasia (MIM
156500)
Lysyl hydroxylase Ehlers-Danlos syndrome type VI (MIM
225400)
ProcollagenN- Ehlers-Danlos syndrome type VII autosomal
proteinase recessive (MIM 225410)
Lysyl hydroxylase Menkes disease (MIM 309400)

Labels: collagen disorders gene defects, ehler danlos syndrome, menkes disease,mutations in
collagen disorders, osteogenesis imperfecta, schmid metaphyseal chondrodysplasia
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SUNDAY, APRIL 26, 2009
35 - AIIMS MAY 2001 biochemistry mcqs with answers

1q) Vitamin K is needed for the post translational
modification of
a) Carboxylation
b) Methylation
c) Hydroxylation
d) Transketolation
2q) Amber codon refers to
a) Initiating codon
b) Mutant codon
c) Stop codon
d) Codon coding for multiple amino acids
3q) At physiological pH, the most stable amino

acid is
a) Histidine
b) Lysine
c) Arginine
d) Leucine
4q) In cystinuria, amino acids excreted are all the

following except:
a) Ornithine
b) Arginine
c) Lysine
d) Histidine
5q) Dietary triglycerides are transported by
a) Chylomicrons
b) LDL
c) VLDL
d) HDL
6q) In which of the following reaction, thiamine

is not used
a) Alpha ketoglutarate to succinyl CoA

b) Glucose to pentose
c) Oxidative decarboxylation of Alpha keto amino acids
d) Lactate to pyruvate
7q) In chromatography, mass movement of the

substances is seen in
a) Electrophoresis
b) Diffusion
c) Osmosis
d) Paper chromatography
8q) The type of chromatography in which

proteins are bound to another substance is
a) Hydrophobic chromatography
b) Affinity chromatography
c) paper chromatography
d) gel chromatography
9q) The end-product of citric acid cycle used in

detoxification of ammonia in brain is
a) Oxaloacetate
b) Alpha keto glutarate
c) Succinate
d) Citrate
To view all the Mcqs of AIIMS MAY 2001 paper with answers click here

Labels: aiims may 2001 biochemistry mcqs with answers, aiims may 2001 complete paper mcqs with
answers, chromatography mcqs, chylomicrons mcqs,citric acid cycle mcqs, cystinuria mcqs, vitamin
k mcqs
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TUESDAY, MARCH 31, 2009
34 - AIIMS november 2002 biochemistry mcqs

1q: an alfa helix of a protein is most likely to be disrupted if a missense
mutation introduces the following aminoacid with in the alpha helical
structure ?
a. alanine
b. aspartic acid
c. tyrosine
d. glycine
explanation : alfa helix of a protein can be disrupted due to the introduction of

the following aminoacids :
1. proline : the imino group of proline is not geometrically compatible with the
right handed spiral of the alpha-helix
2. charged aminoacids : glutamate ,aspartate, lysine, arginine and histidine .
3. aminoacids with bulky side chains : tryptophan
4. aminoacids that branch at the beta-carbon : valine and isoleucine .

33 - AIIMS november 2008 biochemsitry questions
1q: dna without introns is?
A. B dna
b. Z dna
c. C dna
d. Mitochondrial dna
Answer:
2q: all are true about glutathione except?
A. Converts hemoglobin to methemoglobin

b. Decreases free radicals
c. Helps in conjugation reaction
d. Co factor of various enzymes
Answer:
3q: Ribosome has followin enzymatic activity?
A. Peptidyl transferase
b. Amino acyl t rna synthetase
c. Peptidase
d.
Answer:
4q: which enzyme is responsible for carboxylation reaction?
A. Biotin
b.
C.
D. Thiamine pyrophosphate
Answer:
5q: Glowing of firefly is due to?
A. Atp
b. Nadh
c. Gtp
d. Phospho creatinine
Answer:
6q: in carboxylation of clotting factors by vit k which amino acid is

carboxylated?
A. aspartate
b. Glutamate
c. Histamine
d. Histidine
answer:
7q: synthesis of a immunoglobulin in membrane bound or independent form is

determined by?
A. One turn two turn joining rule

b. Allelic exclusion
c. Class switching
d. Differential rna processing
answer:
8q: phosphorlase b is inhibited by?
A. Atp
b. Amp
c. Glucose
d. Calcium
Answer:
9q: In metabolism of xenobiotics all of the followin reactions occour in phase

one except?
A. Conjugation
b. Reduction
c. Hydrolysis
d.oxidation
Answer:
10q: Which is not a second messenger?
A. Amp
b. Guanyl cyclase
c. Dag
d. Ip3
answer:
11q: structure of proteins can be determined by all except?
A. Mass spectrometry
b. Nmr spectrometry
c. Hplc
d.
Answer:
12q: Functions of thiamine?
A. Co enzyme of pyruvate dehydrogenase and alpha keto dehydrogenase

b. Co enzyme of trans ketolase
c.
D.
Answer:
13q: replacing alanine by which amino acid will increase the absorbance of
proteins at 280nm?
A. Leucine
b.tryptophan
c. Proline
d. Arginine
Answer:
Labels: aiims biochemistry mcqs, aiims immunology mcqs, aiims immunology november 2008
mcqs, AIIMS november 2008 biochemistry mcqs, aiims november 2008 paper, aiims past papers
biochemistry mcqs
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WEDNESDAY, NOVEMBER 12, 2008
32 - AIIMS novemeber 2008 biochemistry mcqs - 1

Q1. what is the source of energy for light production in the firefly ?”
1. ATP
2. GTP
3. phosho creatine
4. NADH
The answer is ATP….
Let me start the discussion with this question …..“DO YOU THINK THAT THE
QUESTION IS RELEVANT TO US ?”
If you think this question is irrelevant , then go through the following discussion
& then you ll understand why these AIIMS people want us to know about the
metabolic machinery of this humble organism.
The firefly (Photinus pyralis) contains an enzyme called LUCIFERASE. This enzyme
acts on its substrate – luciferin in the presence of ATP and calcium. The reaction
produces visible light. This reaction has been exploited by the scientific
community for research on
1. antibiotic susceptibility testing

2. pyro sequencing
3. studies of metabolic pathways
4. tumorigenesis
5. transcriptional activity
6. others
ANTI BIOTIC SUSCEPTIBILITY TESTING
Eg : MDR TB
MDR TB is characterized by the presence of resistance to the two drugs
Isoniazide …….. due to mutations of genes Kat G & INH A
Rifampicin……..due the mutations in the gene rpo B gene
To the mycobacteria cultured from the patient`s clinical samples we add adeno
virus carrying the FFLUX gene (which codes for luciferase). Within a few minutes
the mycobacteria exhibit bioluminescence i.e they emit light. Now add the
antibiotic , say INH. If the mycobacteria are sensitive , they soon become non
viable & their metabolic machinery is paralysed and hence ATP is depleted. In the
absence of ATP , the light production is reduced. In fact the degree of reduction
of the bio luminescence can be taken as an indicator of the potency of the drug .
If the strains are resistant , there is NO reduction in light production
PYRO SEQUENCING
This is a type of DNA sequencing.

Take the template DNA which is to be sequenced. To this we add luciferase,
luciferin, calcium , DNA polymerase, ATP sulfurylase
Then we add the deoxy nucleotides (A, C , G , T) one by one and wash them off.
For instance let the d-NTP in x position of the template be A. When we add A, G ,
C there will be no reaction. But when we add T , base pairing occurs , releasing a
PYRO PHOSPHATE, which is acted upon by ATP sulfurylase to produce ATP.
When there is ATP , luciferase is activated and there is luminescence. So we know
that the template contains the d NTP which is complementary to the one that we
added.in the same way the entire NA can be sequenced
STUDYING METABOLIC PATHWAYS
Coenorhabditis elegans (a nematode) was the FIRST MULTICELLULAR ORGANISM

FOR WHICH THE ENTIRE GENOME WAS SEQUENCED.
An interesting aspect is that its cuticular layer is transparent. We can add the
luciferase gene to its genome with the help of adeno or lenti virus. So whenever
its metabolic machinery is activated an ATP is produced, light is emitted.
Likewise whenever a metabolic poison is given the ‘light is switched off’
The degree of bio luminescence is proportional to the amount of ATP present
STUDYING TRANSCRIPTIONAL ACTIVITIES
To study the transcriptional activity of a gene , incorporate the luciferase gene

close to the index gene `s promoter
Whenever there is transcription, luciferase is also activated and light is emitted
TUMORIGENESIS MODELS
The tumor cells tagged with luciferase gene become luminescent and thereby
their growth and susceptibility to chemo therapeutic agents can be studied in
vivo within mouse models
OTHERS
Luciferase gene is also used in forensic medicine - detect blood and in studying
specific cell lines
Labels: aiims biochemistry mcqs, AIIMS november 2008 biochemistry mcqs,aiims november 2008
paper, ATP mcqs, fire fly light production mechanism,luciferase, photinus pyralis
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MONDAY, AUGUST 18, 2008
31 - Rate limiting enzymes of biochemical pathways

RATE LIMITING ENZYMES OF DIFFERENT BIOCHEMICAL PATHWAYS
1. GLYCOLYSIS –-------------------------- phosphofructokinase
2. GLYCOGEN SYNTHESIS –---------- glycogen synthetase
3. GLYCOGENOLYSIS –----------------- glycogen phosphorylase
4. TCA cycle –-------------------------------- isocitrate dehydrogenase
5. CHOLESTEROL SYNTHESIS –----- HMG CoA reductase
6. KETONE BODY SYNTHESIS –------ HMG CoA synthetase
7. UREA SYNTHESIS --------------------– carbamoyl transferase
( ornithine transcarbamoylase )
8. FATTY ACID SYNTHESIS –---------- acetyl coA carboxylase
9. BILE ACID SYNTHESIS –------------- 7 alpha hydroxylase
10. CATECHOLAMINE SYNTHESIS –- tyrosine hydroxylase

Labels: 7 alpha hydroxylase, acetyl coa carboxylase, bile acid synthesis,biochemistry
mcqs, catecholamine synthesis, fatty acid synthesis, rate limiting enzymes mcqs, tyrosine
hydroxylase
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30 - eicosanoid synthesis - PGs , LTs , TXA synthesis
source : www.wikipedia.org
Labels: arachidonic acid metabolism, cox-1, cox-2, cyclooxygenase, ecosanoid synthesis, leukotriene
synthesis, lipoxygenase, prostacyclin synthesis,prostaglandin synthesis, thromboxane synthesis
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SUNDAY, AUGUST 10, 2008
29 - cholesterol synthesis
Cholesterol is a lipid found in the cell membranes of all animal tissues, and is
transported in the blood plasma of all animals. Cholesterol is also a sterol (a
combination steroid and alcohol). Because cholesterol is synthesized by all
eukaryotes, trace amounts of cholesterol are also found in membranes of
plants and fungi.
The name originates from the Greek chole- (bile) and stereos(solid), and the
chemical suffix -ol for an alcohol, as researchers first identified cholesterol in
solid form in gallstones by François Poulletier de la Salle in 1769. However, it is
only in 1815 that chemist Eugène Chevreul named the compound
"cholesterine".
Most of the cholesterol in the body is synthesized by the body and some has
dietary origin. Cholesterol is more abundant in tissues which either synthesize
more or have more abundant densely-packed membranes, for example, the
liver, spinal cord and brain. It plays a central role in many biochemical
processes, such as the composition of cell membranes and the synthesis of
steroid hormones.
Since cholesterol is insoluble in blood, it is transported in the circulatory
system within lipoproteins, complex spherical particles which have an exterior
composed mainly of water-soluble proteins; fats and cholesterol are carried
internally. There is a large range of lipoproteins within blood, generally called,
from larger to smaller size: chylomicrons, very low density lipoprotein (VLDL),
intermediate density lipoprotein (IDL), low density lipoprotein (LDL) and high
density lipoprotein (HDL). The cholesterol within all the various lipoproteins
is identical.
According to the lipid hypothesis, abnormally high cholesterol levels
(hypercholesterolemia), or, more correctly, higher concentrations of LDL and
lower concentrations of functional HDL are strongly associated with
cardiovascular disease because these promote atheroma development in
arteries (atherosclerosis). This disease process leads to myocardial infarction
(heart attack), stroke and peripheral vascular disease. Since higher blood LDL,
especially higher LDL particle concentrations and smaller LDL particle size,
contribute to this process more than the cholesterol content of the LDL
particles [4], LDL particles are often termed "bad cholesterol" because they
have been linked to atheroma formation. On the other hand, high
concentrations of functional HDL, which can remove cholesterol from cells
and atheroma, offer protection and are sometimes referred to colloquially as
"good cholesterol". These balances are mostly genetically determined but can
be changed by body build, medications, food choices and other factors.
Labels: acetyl coa metabolism, bisphosphonates, cholesterol synthesis flowchart,hmg coa

reductase, hmg coa synthetase, lanosterol, mevalonate, squalene,statins, thiolase
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TUESDAY, JUNE 10, 2008
28 - urea cycle
Reactions of cycle:
Ste Catalyze
Reactant Product
p d by
carbamoyl
2ATP + HCO3- +
1M phosphate + 2ADP CPS
NH4+
+ Pi
carbamoyl
2M phosphate + citrulline + Pi OTC
ornithine
citrulline + argininosuccinate
3C ASS
aspartate + ATP + AMP + PPi
4C argininosuccinate Arg + fumarate ASL
5C Arg + H2O ornithine + urea ARG1
CPS1 stands for carbamoyl phosphate synthase 1 enzyme. OTC stands for
ornithine transcarbamoylase. ASS stands for arginine succinate synthase. ASL
stands for arginosuccinate lyase . ARG stands for arginase .the first two steps
occur in mitochondria(M) and the next 3 steps occur in cytosol(C)

Labels: arginosuccinate synthase, CPS 1, kreb-hanseliet urea cycle, ornithine cycle, OTC
enzyme, reactions in mitochondria, steps of urea cycle in mitochondria and cytosol, urea cycle
Reaction
s:
THURSDAY, MAY 29, 2008
27 - polymerase chain reaction ( PCR )

check out this page for an animation which excellently depicts the
basic principle and mechanism underlying PCR . CLICK HERE
The polymerase chain reaction (PCR) is a technique widely used in

molecular biology. It derives its name from one of its key components, a
DNA polymerase used to amplify a piece of DNA by in vitro enzymatic
replication. As PCR progresses, the DNA thus generated is itself used as
template for replication. This sets in motion a chain reaction in which the
DNA template is exponentially amplified. With PCR it is possible to amplify
a single or few copies of a piece of DNA across several orders of magnitude,
generating millions or more copies of the DNA piece. PCR can be
extensively modified to perform a wide array of genetic manipulations.
Almost all PCR applications employ a heat-stable DNA polymerase, such as Taq
polymerase, an enzyme originally isolated from the bacterium Thermus
aquaticus. This DNA polymerase enzymatically assembles a new DNA strand
from DNA building blocks, the nucleotides, using single-stranded DNA as
template and DNA oligonucleotides (also called DNA primers) required for
initiation of DNA synthesis. The vast majority of PCR methods use thermal
cycling, i.e., alternately heating and cooling the PCR sample to a defined series
of temperature steps. These thermal cycling steps are necessary to physically
separate the strands (at high temperatures) in a DNA double helix (DNA
melting) used as template during DNA synthesis (at lower temperatures) by the
DNA polymerase to selectively amplify the target DNA. The selectivity of PCR
results from the use of primers that are complementary to the DNA region
targeted for amplification under specific thermal cycling conditions.
Developed in 1983 by Kary Mullis,[1] PCR is now a common and often
indispensable technique used in medical and biological research labs for a
variety of applications.[2][3] These include DNA cloning for sequencing, DNA-
based phylogeny, or functional analysis of genes; the diagnosis of hereditary
diseases; the identification of genetic fingerprints (used in forensics and
paternity testing); and the detection and diagnosis of infectious diseases. In
1993 Mullis won the Nobel Prize for his work on PCR.[4]
PCR PRINCIPLE
PCR is used to amplify specific regions of a DNA strand (the DNA target). This
can be a single gene, a part of a gene, or a non-coding sequence. Most PCR
methods typically amplify DNA fragments of up to 10 kilo base pairs (kb),
although some techniques allow for amplification of fragments up to 40 kb in
size.[5]
A basic PCR set up requires several components and reagents.[6]These
components include:
• DNA template that contains the DNA region (target) to be amplified.
• One or more primers, which are complementary to the DNA regions at
the 5' (five prime) and 3' (three prime) ends of the DNA region.
• A DNA polymerase such as Taq polymerase or another DNA polymerase
with a temperature optimum at around 70°C.
• Deoxynucleoside triphosphates (dNTPs; also very commonly and
erroneously called deoxynucleotide triphosphates), the building blocks from
which the DNA polymerases synthesizes a new DNA strand.
• Buffer solution, providing a suitable chemical environment for optimum
activity and stability of the DNA polymerase.
• Divalent cations, magnesium or manganese ions; generally Mg2+ is used,
but Mn2+ can be utilized for PCR-mediated DNA mutagenesis, as higher
Mn2+concentration increases the error rate during DNA synthesis[7]
• Monovalent cation potassium ions.
The PCR is commonly carried out in a reaction volume of 15-100 μl in small
reaction tubes (0.2-0.5 ml volumes) in a thermal cycler. The thermal cycler
heats and cools the reaction tubes to achieve the temperatures required at each
step of the reaction (see below). Many modern thermal cyclers make use of the
Peltier effect which permits both heating and cooling of the block holding the
PCR tubes simply by reversing the electric current. Thin-walled reaction tubes
permit favorable thermal conductivity to allow for rapid thermal equilibration.
Most thermal cyclers have heated lids to prevent condensation at the top of the
reaction tube. Older thermocyclers lacking a heated lid require a layer of oil on
top of the reaction mixture or a ball of wax inside the tube.
Procedure
The PCR usually consists of a series of 20 to 35 repeated temperature
changes called cycles; each cycle typically consists of 2-3 discrete
temperature steps. Most commonly PCR is carried out with cycles that
have three temperature steps (Fig. 2). The cycling is often preceded by
a single temperature step (called hold) at a high temperature (>90°C),
and followed by one hold at the end for final product extension or
brief storage. The temperatures used and the length of time they are
applied in each cycle depend on a variety of parameters. These
include the enzyme used for DNA synthesis, the concentration of
divalent ions and dNTPs in the reaction, and the melting temperature
(Tm) of the primers.[8]
• Initialization step: This step consists of heating the reaction to a
temperature of 94-96°C (or 98°C if extremely thermostable polymerases are
used), which is held for 1-9 minutes. It is only required for DNA polymerases
that require heat activation by hot-start PCR.[9]
• Denaturation step: This step is the first regular cycling event and
consists of heating the reaction to 94-98°C for 20-30 seconds. It causes melting
of DNA template and primers by disrupting the hydrogen bonds between
complementary bases of the DNA strands, yielding single strands of DNA.
• Annealing step: The reaction temperature is lowered to 50-65°C for 20-
40 seconds allowing annealing of the primers to the single-stranded DNA
template. Typically the annealing temperature is about 3-5 degrees Celsius
below the Tm of the primers used. Stable DNA-DNA hydrogen bonds are only
formed when the primer sequence very closely matches the template sequence.
The polymerase binds to the primer-template hybrid and begins DNA
synthesis.
• Extension/elongation step: The temperature at this step depends on the
DNA polymerase used; Taq polymerase has its optimum activity temperature
at 75-80°C,[10][11]and commonly a temperature of 72°C is used with this enzyme.
At this step the DNA polymerase synthesizes a new DNA strand complementary
to the DNA template strand by adding dNTPs that are complementary to the
template in 5' to 3' direction, condensing the 5'-phosphate group of the dNTPs
with the 3'-hydroxyl group at the end of the nascent (extending) DNA strand.
The extension time depends both on the DNA polymerase used and on the
length of the DNA fragment to be amplified. As a rule-of-thumb, at its optimum
temperature, the DNA polymerase will polymerize a thousand bases per
minute. Under optimum conditions, i.e., if there are no limitations due to
limiting substrates or reagents, at each extension step, the amount of DNA
target is doubled, leading to exponential (geometric) amplification of the
specific DNA fragment.
• Final elongation: This single step is occasionally performed at a
temperature of 70-74°C for 5-15 minutes after the last PCR cycle to ensure that
any remaining single-stranded DNA is fully extended.
• Final hold: This step at 4-15°C for an indefinite time may be employed for
short-term storage of the reaction.
Figure 3: Ethidium bromide-stained PCR products after gel
electrophoresis. Two sets of primers were used to amplify a target
sequence from three different tissue samples. No amplification is
present in sample #1; DNA bands in sample #2 and #3 indicate
successful amplification of the target sequence. The gel also shows a
positive control, and a DNA ladder containing DNA fragments of
defined length for sizing the bands in the experimental PCRs.
To check whether the PCR generated the anticipated DNA fragment (also
sometimes referred to as the amplimer or amplicon), agarose gel
electrophoresis is employed for size separation of the PCR products. The
size(s) of PCR products is determined by comparison with a DNA ladder (a
molecular weight marker), which contains DNA fragments of known size, run
on the gel alongside the PCR products (see Fig. 3).
PCR STAGES
The PCR process can be divided into three stages:
Exponential amplification: At every cycle, the amount of product is doubled
(assuming 100% reaction efficiency). The reaction is very specific and precise.
[citation needed]
Levelling off stage: The reaction slows as the DNA polymerase loses activity
and as consumption of reagents such as dNTPs and primers causes them to
become limiting.
Plateau: No more product accumulates due to exhaustion of reagents and
enzyme.
PCR optimization
In practice, PCR can fail for various reasons, in part due to its sensitivity to
contamination causing amplification of spurious DNA products. Because of
this, a number of techniques and procedures have been developed for
optimizing PCR conditions.[12][13]Contamination with extraneous DNA is
addressed with lab protocols and procedures that separate pre-PCR mixtures
from potential DNA contaminants.[6] This usually involves spatial separation of
PCR-setup areas from areas for analysis or purification of PCR products, and
thoroughly cleaning the work surface between reaction setups. Primer-design
techniques are important in improving PCR product yield and in avoiding the
formation of spurious products, and the usage of alternate buffer components
or polymerase enzymes can help with amplification of long or otherwise
problematic regions of DNA.
APPLICATION OF PCR
Isolation of genomic DNA

PCR allows isolation of DNA fragments from genomic DNA by selective
amplification of a specific region of DNA. This use of PCR augments many
methods, such as generating hybridization probes for Southern or northern
hybridization and DNA cloning, which require larger amounts of DNA,
representing a specific DNA region. PCR supplies these techniques with high
amounts of pure DNA, enabling analysis of DNA samples even from very small
amounts of starting material.
Other applications of PCR include DNA sequencing to determine unknown
PCR-amplified sequences in which one of the amplification primers may be
used in Sanger sequencing, isolation of a DNA sequence to expedite
recombinant DNA technologies involving the insertion of a DNA sequence into
a plasmid or the genetic material of another organism. Bacterial colonies
(E.coli) can be rapidly screened by PCR for correct DNA vector constructs[14].
PCR may also be used for genetic fingerprinting; a forensic technique used to
identify a person or organism by comparing experimental DNAs through
different PCR-based methods.
Some PCR 'fingerprints' methods have high discriminative power and can be
used to identify genetic relationships between individuals, such as parent-child
or between siblings, and are used in paternity testing (Fig. 4). This technique
may also be used to determine evolutionary relationships among organisms.
Amplification and quantitation of DNA
Because PCR amplifies the regions of DNA that it targets, PCR can be used to
analyze extremely small amounts of sample. This is often critical for forensic
analysis, when only a trace amount of DNA is available as evidence. PCR may
also be used in the analysis of ancient DNA that is thousands of years old.
These PCR-based techniques have been successfully used on animals, such as a
forty-thousand-year-old mammoth, and also on human DNA, in applications
ranging from the analysis of Egyptian mummies to the identification of a
Russian Tsar.[15]
Quantitative PCR methods allow the estimation of the amount of a given
sequence present in a sample – a technique often applied to quantitatively
determine levels of gene expression. Real-time PCR is an established tool for
DNA quantification that measures the accumulation of DNA product after each
round of PCR amplification.
PCR in diagnosis of diseases
PCR allows early diagnosis of malignant diseases such as leukemia and
lymphomas, which is currently the highest developed in cancer research and is
already being used routinely.[citation needed]PCR assays can be performed directly
on genomic DNA samples to detect translocation-specific malignant cells at a
sensitivity which is at least 10,000 fold higher than other methods.[citation needed]
PCR also permits identification of non-cultivatable or slow-growing
microorganisms such as mycobacteria, anaerobic bacteria, or viruses from
tissue culture assays and animal models. The basis for PCR diagnostic
applications in microbiology is the detection of infectious agents and the
discrimination of non-pathogenic from pathogenic strains by virtue of specific
genes.[citation needed]
Viral DNA can likewise be detected by PCR. The primers used need to be specific to the
targeted sequences in the DNA of a virus, and the PCR can be used for diagnostic analyses
or DNA sequencing of the viral genome. The high sensitivity of PCR permits virus detection
soon after infection and even before the onset of disease. Such early detection may give
physicians a significant lead in treatment. The amount of virus ("viral load") in a patient can
also be quantified by PCR-based DNA quantitation techniques (see below).
Variations on the basic PCR technique
• Allele-specific PCR: This diagnostic or cloning technique is used to
identify or utilize single-nucleotide polymorphisms (SNPs) (single base
differences in DNA). It requires prior knowledge of a DNA sequence, including
differences between alleles, and uses primers whose 3' ends encompass the
SNP. PCR amplification under stringent conditions is much less efficient in the
presence of a mismatch between template and primer, so successful
amplification with an SNP-specific primer signals presence of the specific SNP
in a sequence.[16] See SNP genotyping for more information.
• Assembly PCR or Polymerase Cycling Assembly (PCA): Assembly PCR is
the artificial synthesis of long DNA sequences by performing PCR on a pool of
long oligonucleotides with short overlapping segments. The oligonucleotides
alternate between sense and antisense directions, and the overlapping
segments determine the order of the PCR fragments thereby selectively
producing the final long DNA product.[17]
• Asymmetric PCR: Asymmetric PCR is used to preferentially amplify one
strand of the original DNA more than the other. It finds use in some types of
sequencing and hybridization probing where having only one of the two
complementary stands is required. PCR is carried out as usual, but with a great
excess of the primers for the chosen strand. Due to the slow (arithmetic)
amplification later in the reaction after the limiting primer has been used up,
extra cycles of PCR are required.[18] A recent modification on this process,
known as Linear-After-The-Exponential-PCR (LATE-PCR), uses a limiting
primer with a higher melting temperature (Melting temperature|Tm) than the
excess primer to maintain reaction efficiency as the limiting primer
concentration decreases mid-reaction.[19]
• Helicase-dependent amplification: This technique is similar to
traditional PCR, but uses a constant temperature rather than cycling through
denaturation and annealing/extension cycles. DNA Helicase, an enzyme that
unwinds DNA, is used in place of thermal denaturation.[20]
• Hot-start PCR: This is a technique that reduces non-specific
amplification during the initial set up stages of the PCR. The technique may be
performed manually by heating the reaction components to the melting
temperature (e.g., 95˚C) before adding the polymerase.[21] Specialized enzyme
systems have been developed that inhibit the polymerase's activity at ambient
temperature, either by the binding of an antibody[9] or by the presence of
covalently bound inhibitors that only dissociate after a high-temperature
activation step. Hot-start/cold-finish PCR is achieved with new hybrid
polymerases that are inactive at ambient temperature and are instantly
activated at elongation temperature.
• Intersequence-specific (ISSR) PCR: a PCR method for DNA
fingerprinting that amplifies regions between some simple sequence repeats to
produce a unique fingerprint of amplified fragment lengths.[22]
• Inverse PCR: a method used to allow PCR when only one internal
sequence is known. This is especially useful in identifying flanking sequences
to various genomic inserts. This involves a series of DNA digestions and self
ligation, resulting in known sequences at either end of the unknown sequence.
[23]
• Ligation-mediated PCR: This method uses small DNA linkers ligated to
the DNA of interest and multiple primers annealing to the DNA linkers; it has
been used for DNA sequencing, genome walking, and DNA footprinting.[24]
• Methylation-specific PCR (MSP): The MSP method was developed by
Stephen Baylin and Jim Herman at theJohns Hopkins School of Medicine,
[25]
and is used to detect methylation of CpG islands in genomic DNA. DNA is
first treated with sodium bisulfite, which converts unmethylated cytosine
bases to uracil, which is recognized by PCR primers as thymine. Two PCRs are
then carried out on the modified DNA, using primer sets identical except at any
CpG islands within the primer sequences. At these points, one primer set
recognizes DNA with cytosines to amplify methylated DNA, and one set
recognizes DNA with uracil or thymine to amplify unmethylated DNA. MSP
using qPCR can also be performed to obtain quantitative rather than
qualitative information about methylation.
• Miniprimer PCR: Miniprimer PCR uses a novel thermostable polymerase
(S-Tbr) that can extend from short primers ("smalligos") as short as 9 or 10
nucleotides, instead of the approximately 20 nucleotides required by Taq. This
method permits PCR targeting smaller primer binding regions, and is
particularly useful to amplify unknown, but conserved, DNA sequences, such
as the 16S (or eukaryotic 18S) rRNA gene. 16S rRNA miniprimer PCR was used
to characterize a microbial mat community growing in an extreme
environment, a hypersaline pond in Puerto Rico. In that study, deeply
divergent sequences were discovered with high frequency and included
representatives that deﬁned two new division-level taxa, suggesting that
miniprimer PCR may reveal new dimensions of microbial diversity.[26] By
enlarging the "sequence space" that may be queried by PCR primers, this
technique may enable novel PCR strategies that are not possible within the
limits of primer design imposed by Taq and other commonly used enzymes.
• Multiplex Ligation-dependent Probe Amplification (MLPA): permits
multiple targets to be amplified with only a single primer pair, thus avoiding
the resolution limitations of multiplex PCR (see below).
• Multiplex-PCR: The use of multiple, unique primer sets within a single
PCR mixture to produce amplicons of varying sizes specific to different DNA
sequences. By targeting multiple genes at once, additional information may be
gained from a single test run that otherwise would require several times the
reagents and more time to perform. Annealing temperatures for each of the
primer sets must be optimized to work correctly within a single reaction, and
amplicon sizes, i.e., their base pair length, should be different enough to form
distinct bands when visualized by gel electrophoresis.
• Nested PCR: increases the specificity of DNA amplification, by reducing
background due to non-specific amplification of DNA. Two sets of primers are
being used in two successive PCRs. In the first reaction, one pair of primers is
used to generate DNA products, which besides the intended target, may still
consist of non-specifically amplified DNA fragments. The product(s) are then
used in a second PCR with a set of primers whose binding sites are completely
or partially different from and located 3' of each of the primers used in the first
reaction. Nested PCR is often more successful in specifically amplifying long
DNA fragments than conventional PCR, but it requires more detailed
knowledge of the target sequences.
• Overlap-extension PCR: is a genetic engineering technique allowing the
construction of a DNA sequence with an alteration inserted beyond the limit of
the longest practical primer length.
• Quantitative PCR (Q-PCR): is used to measure the quantity of a PCR
product (preferably real-time). It is the method of choice to quantitatively
measure starting amounts of DNA, cDNA or RNA. Q-PCR is commonly used to
determine whether a DNA sequence is present in a sample and the number of
its copies in the sample. The method with currently the highest level of
accuracy is Quantitative real-time PCR. It is often confusingly known as RT-
PCR (Real Time PCR) or RQ-PCR. QRT-PCR or RTQ-PCR are more appropriate
contractions. RT-PCR commonly refers to reverse transcription PCR (see
below), which is often used in conjunction with Q-PCR. QRT-PCR methods use
fluorescent dyes, such as Sybr Green, or fluorophore-containing DNA probes,
such as TaqMan, to measure the amount of amplified product in real time.
• RT-PCR: (Reverse Transcription PCR) is a method used to amplify,
isolate or identify a known sequence from a cellular or tissue RNA. The PCR is
preceded by a reaction using reverse transcriptase to convert RNA to cDNA.
RT-PCR is widely used in expression profiling, to determine the expression of a
gene or to identify the sequence of an RNA transcript, including transcription
start and termination sites and, if the genomic DNA sequence of a gene is
known, to map the location of exons and introns in the gene. The 5' end of a
gene (corresponding to the transcription start site) is typically identified by an
RT-PCR method, named RACE-PCR, short for Rapid Amplification of cDNA
Ends.
• Solid Phase PCR: encompasses multiple meanings, including Polony
Amplification (where PCR colonies are derived in a gel matrix, for example),
'Bridge PCR' (the only primers present are covalently linked to solid support
surface), conventional Solid Phase PCR (where Asymmetric PCR is applied in
the presence of solid support bearing primer with sequence matching one of
the aqueous primers) and Enhanced Solid Phase PCR[27](where conventional
Solid Phase PCR can be improved by employing high Tm solid support primer
with application of a thermal 'step' to favour solid support priming).
• TAIL-PCR: Thermal asymmetric interlaced PCR is used to isolate
unknown sequence flanking a known sequence. Within the known sequence
TAIL-PCR uses a nested pair of primers with differing annealing temperatures;
a degenerate primer is used to amplify in the other direction from the
unknown sequence.[28]
• Touchdown PCR: a variant of PCR that aims to reduce nonspecific
background by gradually lowering the annealing temperature as PCR cycling
progresses. The annealing temperature at the initial cycles is usually a few
degrees (3-5˚C) above the Tm of the primers used, while at the later cycles, it is
a few degrees (3-5˚C) below the primer Tm. The higher temperatures give
greater specificity for primer binding, and the lower temperatures permit
more efficient amplification from the specific products formed during the
initial cycles.[29]
• PAN-AC: This method uses isothermal conditions for amplification, and
may be used in living cells.[30][31]
• Universal Fast Walking: this method allows genome walking and genetic
fingerprinting using a more specific 'two-sided' PCR than conventional 'one-
sided' approaches (using only one gene-specific primer and one general primer
- which can lead to artefactual 'noise')[32] by virtue of a mechanism involving
lariat structure formation. Streamlined derivatives of UFW are LaNe RAGE
(lariat-dependent nested PCR for rapid amplification of genomic DNA
ends) [33], 5'RACE LaNe[34] and 3'RACE LaNe [35].
HISTORY
A 1971 paper in the Journal of Molecular Biology by Kleppe and co-workers
first described a method using an enzymatic assay to replicate a short DNA
template with primers in vitro. However, this early manifestation of the basic
PCR principle did not receive much attention, and the invention of the
polymerase chain reaction in 1983 is generally credited to Kary Mullis.[37]
At the core of the PCR method is the use of a suitable DNA polymerase able to
withstand the high temperatures of >90°C (>195°F) required for separation of
the two DNA strands in the DNA double helix after each replication cycle. The
DNA polymerases initially employed for in vitro experiments presaging PCR
were unable to withstand these high temperatures. So the early procedures for
DNA replication were very inefficient, time consuming, and required large
amounts of DNA polymerase and continual handling throughout the process.
A 1976 discovery of Taq polymerase a DNA polymerase purified from the
thermophilic bacterium, Thermus aquaticus, which naturally occurs in hot (50
to 80 °C (120 to 175 °F)) environments paved the way for dramatic
improvements of the PCR method. The DNA polymerase isolated from T.
aquaticus is stable at high temperatures remaining active even after DNA
denaturation,thus obviating the need to add new DNA polymerase after each
cycle. This allowed an automated thermocycler-based process for DNA
amplification.
At the time he developed PCR in 1983, Mullis was working in Emeryville,
California for Cetus Corporation, one of the first biotechnology companies.
There, he was responsible for synthesizing short chains of DNA. Mullis has
written that he conceived of PCR while cruising along the Pacific Coast
Highwayone night in his car. He was playing in his mind with a new way of
analyzing changes (mutations) in DNA when he realized that he had instead
invented a method of amplifying any DNA region through repeated cycles of
duplication driven by DNA polymerase.
In Scientific American, Mullis summarized the procedure: "Beginning with a
single molecule of the genetic material DNA, the PCR can generate 100 billion
similar molecules in an afternoon. The reaction is easy to execute. It requires
no more than a test tube, a few simple reagents, and a source of heat." He was
awarded the Nobel Prize in Chemistry in 1993 for his invention,seven years
after he and his colleagues at Cetus first put his proposal to practice. However,
some controversies have remained about the intellectual and practical
contributions of other scientists to Mullis' work, and whether he had been the
sole inventor of the PCR principle.
Patent wars
The PCR technique was patented by Cetus Corporation, where Mullis worked
when he invented the technique in 1983. The Taqpolymerase enzyme was also
covered by patents. There have been several high-profile lawsuits related to the
technique, including an unsuccessful lawsuit brought by DuPont. The
pharmaceutical company Hoffmann-La Roche purchased the rights to the
patents in 1992 and currently holds those that are still protected.
A related patent battle over the Taq polymerase enzyme is still
ongoing in several jurisdictions around the world between Roche and
Promega. The legal arguments have extended beyond the life of the
original PCR and Taq polymerase patents, which expired on March 28,
2005
Labels: dna amplification, DNA polymerases, kary mullis, PCR, pfu polymerase,polymerase chain
reaction, real time pcr, taq polymerase, use of pcr in forensic sciences
Reaction
s:
TUESDAY, MAY 27, 2008
24 - biochemistry mcqs - 156 to 166

156. which of the following require template for its formation ?
a- carbohydrate
b- protein
c- lipids
d- phospholipids
e- nucleic acids
answer : b and e . proteins require the m RNA template where as the
nucleic acids require the DNA template .
157. which of the following are intermediate metabolites in TCA cycle ?
a- pyruvate
b- malonate
c- oxaloacetate
d- isocitrate
e- nitric oxide
answers : c and d .
158. which is the smallest fundamental unit coding for DNA synthesis ?
a- cistron
b- operon
c- replican
d- anti codon
answer : a . cistron .
159. metabolic bone disease is caused by excess intake of which vitamin ?
a- vitamin A
b- vitamin B
c- vitamin C
d- vitamin D
e- vitamin E
answer : a and d are the answers . both vitamin D and vitamin A .
160. hypolipidemic agents act on :
a- HMG COA synthetase
b- HMG COA oxygenase
c- HMG COA reductase
d- HMG COA hydratase
e- HMG COA mutase
Answer : c. HMG CO A reductase .
161. correct sequence of enzymes required for DNA formation ?
Answer : DNA topoisomerase – RNA polymerase – DNA polymerase 3 – DNA
polymerase 1 – DNA ligase .
I did not give u the options because , it would confuse u further .
162. which of the following are the bile acids synthesized in the liver ?
( primary bile acids )
a- cholic acid
b- chenodeoxy cholic acid
c- deoxycholic acid
d- lithocholic acid
e- taurocholic acid
f- glycocholic acid
answer : a and b are the answers . c and d are the secondary bile acids . e
and f are the bile salts . sodium taurocholate and sodium glycocholate are
the bile salts formed on combining with sodium .
163. vitamin required for the conversion of hydroxy proline to proline ?
a- vitamin C
b- vitamin E
c- pyridoxal phosphate
d- biotin
answer : vitamin C .
164. enzyme activity measured in beri beri is ?
a- transketolase
b- transaminase
c- decarboxylase
d- deaminases
answer : a . transketolase .
165. muscle cannot make use of glycogen for energy because of deficiency
of ?
a- glucokinase
b- phosphoglucomutase
c- glucose 6 phosphatase
d- muscle phosphorylase
answer : c .
166. pompe’s disease is due to deficiency of which enzyme ?
a- branching enzyme
b- glucose 6 phosphatase
c- acid maltase deficiency
d- muscle phosphorylase
answer : c . acid maltase deficiency . acid maltase is otherwise called
alpha glucosidase .
Labels: bile acids mcqs, biochemistry mcqs, biochemistry pgi chandigarh mcqs,citric acid cycle
mcqs, glycogen storage disorders mcqs, primary and secondary bile acids, tca cycle mcqs, vitamin c
mcqs
Reaction
s:
MONDAY, MAY 26, 2008
23 - cytochrome oxidase inhibitors

question : cytochrome oxidase is inhibited by :
a- cyanide
b- aluminium phosphide
c- phenobarbitone
d- carbonmonoxide
e- NO
answer : cyanide and carbon monoxide are the answers .
inhibitors of cytochrome ( a + a3 ) oxidase ( complex IV ) of electron transport

chain ( ETC ) are :
1. carbonmonoxide ( inhibits the enzyme by combining with O2 binding sites )

2. cyanide
3. azide ( sodium azide example )
4. H2S
Labels: azides mcqs, biochemistry pgi chandigarh mcqs, carbonmonoxide mcqs,complex 4 of

etc, cyanide mcqs, cytochrome oxidase inhibitors, electron transport chain mcqs, etc mcqs
Reaction
s:
22 - DNA polymerase mcqs

question : DNA polymerases have
a- 3' - 5' polymerase activity

b- 5' - 3' polymerase activity
c- 3' - 5' exonuclease activity
d- 5' - 3' exonuclease activity
e- endonuclease activity
answer : b , c , d are the answers .
DNA Polymerase 1 has both proof reading and excision repair activity , that is it
has both 3' - 5' exonuclease activity and 5' - 3' exonuclease activity respectively.
where as the DNA polymerase 2 and 3 have only proof reading activity and no
excision repair activity, that is they have 3' - 5' exonuclease activity and no 5' -
3' exonuclease activity.
all the three DNA polymerases have only 5' -3' polymerase activity only.
Labels: 3'-5' exonuclease activity, 5'-3' polymerase activity, DNA polymerase 1,DNA polymerase
2, DNA polymerase 3, DNA polymerase mcqs, DNA replication enzymes, excision repair, genetics
mcqs, proof reading
Reaction
s:
WEDNESDAY, MAY 14, 2008
21 - biochemical pathways and their location

PATHWAYS AND THE ORGANELLES IN WHICH THEY TAKE PLACE :
1. GLYCOLYSIS --------------------- CYTOPLASM

2. T.C.A CYCLE --------------------- MITOCHONDRIA
3. FATTY ACID SYNTHESIS ---- CYTOPLASM
4. FATTY ACID OXIDATION ---- MITOCHONDRIA
--------------------------------------------------------------------------------------------------
-----
5. KETONE BODY SYNTHESIS - MITOCHONDRIA
6. KETONE BODY OXIDATION – MITOCHONDRIA
7. H.M.P SHUNT --------------------- CYTOPLASM
8. GLUCONEOGENESIS ----------- CYTOPLASM AND
MITOCHONDRIA
--------------------------------------------------------------------------------------------------
-----
9. FATTY ACID ELONGATION – S . E . R
10. VERY LONG CHAIN FATTY ACID OXIDATION – PEROXISOME
11. ETHER PHOSPHOLIPID SYNTHESIS – PEROXISOME
12. CHOLESTEROL SYNTHESIS – CYTOPLASM AND ENDOPLASMIC
RETICULUM .
--------------------------------------------------------------------------------------------------
-----
Labels: biochemical pathways and their location, cholesterol synthesis organelle,fatty acid elongation
organelle, gluconeogenesis location, reactions in peroxisomes, tca cycle organelle
Reaction
s:
20 - enzymes with zinc as cofactor
QUESTION : all the following enzymes have zinc as a cofactor except ?
a- glutamate dehydrogenase
b- alcohol dehydrogenase
c- lactate dehydrogenase
d- alkaline phosphatase
e- glutathione peroxidase
answer : e . glutathione peroxidase uses selenium as a cofactor and not zinc.
the list of enzymes which use zinc as a cofactor are :
1. glutamate dehydrogenase
2. alcohol dehydrogenase
3. lactate dehydrogenase
4. carbonic anhydrase
5. alkaline phosphatase
6. DNA polymerase
7. RNA polymerase
8. delta-ALA dehydratase
9. superoxide dismutase
10. pancreatic carboxypeptidase

Labels: alkaline phosphatase, delta-ala dehydratase, enzyme with selenium as cofactor, enzymes with
zinc as cofactor, gdh, glutathione peroxidase, ldh,pancreatic carboxypeptidase
Reaction
s:
THURSDAY, MARCH 6, 2008
19 - vitamin B6 ( pyridoxine )
Vitamin B6
Vitamin B6 is a water-soluble vitamin that was first isolated in the 1930s. There
are three traditionally considerd forms of vitamin B6: pyridoxal (PL),
pyridoxine (PN), pyridoxamine (PM). The phosphate ester derivative pyridoxal
5'-phosphate (PLP) is the principal coenzyme form and has the most
importance in human metabolism .
Function
Vitamin B6 must be obtained from the diet because humans cannot synthesize
it. PLP plays a vital role in the function of approximately 100 enzymes that
catalyze essential chemical reactions in the human body . For example, PLP
functions as a coenzyme for glycogen phosphorylase, an enzyme that catalyzes
the release of glucose from stored glycogen. Much of the PLP in the human
body is found in muscle bound to glycogen phosphorylase. PLP is also a
coenzyme for reactions used to generate glucose from amino acids, a process
known as gluconeogenesis .
Nervous system function
In the brain, the synthesis of the neurotransmitter, serotonin, from the amino
acid, tryptophan, is catalyzed by a PLP-dependent enzyme. Other
neurotransmitters, such as dopamine, norepinephrine and gamma-
aminobutyric acid (GABA), are also synthesized using PLP-dependent enzymes
.
Red blood cell formation and function
PLP functions as a coenzyme in the synthesis of heme, an iron-containing
component of hemoglobin. Hemoglobin is found in red blood cells and is
critical to their ability to transport oxygen throughout the body. Both PL and
PLP are able to bind to the hemoglobin molecule and affect its ability to pick up
and release oxygen. However, the impact of this on normal oxygen delivery to
tissues is not known .
Niacin formation
The human requirement for another B vitamin, niacin, can be met in part by
the conversion of the essential amino acid, tryptophan, to niacin, as well as
through dietary intake. PLP is a coenzyme for a critical reaction in the
synthesis of niacin from tryptophan; thus, adequate vitamin B6 decreases the
requirement for dietary niacin .
Hormone function
Steroid hormones, such as estrogen and testosterone, exert their effects in the
body by binding to steroid hormone receptors in the nucleus of the cell and
altering gene transcription. PLP binds to steroid receptors in a manner that
inhibits the binding of steroid hormones, thus decreasing their effects. The
binding of PLP to steroid receptors for estrogen, progesterone, testosterone,
and other steroid hormones suggests that the vitamin B6 status of an individual
may have implications for diseases affected by steroid hormones, including
breast cancer and prostate cancers .
Nucleic acid synthesis
PLP serves as a coenzyme for a key enzyme involved in the mobilization of
single-carbon functional groups (one-carbon metabolism). Such reactions are
involved in the synthesis of nucleic acids. The effect of vitamin B6 deficiency on
the function of the immune system may be partly related to the role of PLP in
one-carbon metabolism (see Disease Prevention).
Deficiency
Severe deficiency of vitamin B6 is uncommon. Alcoholics are thought to be
most at risk of vitamin B6 deficiency due to low dietary intakes and impaired
metabolism of the vitamin. In the early 1950s, seizures were observed in
infants as a result of severe vitamin B6 deficiency caused by an error in the
manufacture of infant formula. Abnormal electroencephalogram (EEG)
patterns have been noted in some studies of vitamin B6 deficiency. Other
neurologic symptoms noted in severe vitamin B6 deficiency include irritability,
depression, and confusion; additional symptoms include inflammation of the
tongue, sores or ulcers of the mouth, and ulcers of the skin at the corners of the
mouth .
The Recommended Dietary Allowance (RDA)
Because vitamin B6 is involved in many aspects of metabolism, several factors
are likely to effect an individual's requirement for vitamin B6. Of those factors,
protein intake has been the most studied. Increased dietary protein results in
an increased requirement for vitamin B6, probably because PLP is a coenzyme
for many enzymes involved in amino acid metabolism . Unlike previous
recommendations, the Food and Nutrition Board (FNB) of
the Institute of Medicine did not express the most recent RDA for vitamin B6 in
terms of protein intake, although the relationship was considered in setting the
RDA . The current RDA was revised by the Food and Nutrition Board (FNB) in
1998 and is presented in the table below.
Recommended Dietary Allowance (RDA) for Vitamin B6
Life Stage Age Males (mg/day) Females (mg/day)
Infants 0-6 months 0.1 (AI) 0.1 (AI)
Infants 7-12 months 0.3 (AI) 0.3 (AI)
Children 1-3 years 0.5 0.5

Adolescents 14-18 years 1.3 1.2
Adults 19-50 years 1.3 1.3
Adults 51 years and older 1.7 1.5
Pregnancy all ages - 1.9
Breast-feeding all ages - 2.0
Disease Prevention
Homocysteine and cardiovascular disease
Even moderately elevated levels of homocysteine in the blood have been
associated with increased risk for cardiovascular disease, including heart
disease and stroke . During protein digestion, amino acids, including
methionine, are released. Homocysteine is an intermediate in the metabolism
of methionine. Healthy individuals utilize two different pathways to metabolize
homocysteine. One pathway converts homocysteine back to methionine and is
dependent on folic acid and vitamin B12. The other pathway converts
homocysteine to the amino acid cysteine and requires two vitamin B6(PLP)-
dependent enzymes. Thus, the amount of homocysteine in the blood is
regulated by at least three vitamins: folic acid, vitamin B12, and vitamin
B6 (diagram). Several large observational studies have demonstrated an
association between low vitamin B6 intake or status with increased blood
homocysteine levels and increased risk of cardiovascular diseases. A large
prospective study found the risk of heart disease in women who consumed, on
average, 4.6 mg of vitamin B6 daily was only 67% of the risk in women who
consumed an average of 1.1 mg daily . Another large prospective study found
higher plasma levels of PLP were associated with a decreased risk of
cardiovascular disease independent of homocysteine levels . Further, several
studies have reported that low plasma PLP status is a risk factor for coronary
artery disease . In contrast to folic acid supplementation, studies
supplementing individuals with only vitamin B6 have not resulted in significant
decreases in basal (fasting) levels of homocysteine. However, one study found
that vitamin B6 supplementation was effective in lowering blood homocysteine
levels after an oral dose of methionine (methionine load test) was given ,
suggesting vitamin B6 may play a role in the metabolism of homocysteine after
meals.
Immune function
Low vitamin B6 intake and nutritional status have been associated with
impaired immune function, especially in the elderly. Decreased production of
immune system cells known as lymphocytes, as well as decreased production
of an important immune system protein called interleukin-2, have been
reported in vitamin B6 deficient individuals . Restoration of vitamin B6 status
has resulted in normalization of lymphocyte proliferation and interleukin-2
production, suggesting that adequate vitamin B6intake is important for optimal
immune system function in older individuals . However, one study found that
the amount of vitamin B6 required to reverse these immune system
impairments in the elderly was 2.9 mg/day for men and 1.9 mg/day for women;
these vitamin B6 requirements are higher than the current RDA .
Cognitive function
A few studies have associated cognitive decline in the elderly or Alzheimer's
disease with inadequate nutritional status of folic acid, vitamin B12, and
vitamin B6 and thus, elevated levels of homocysteine . One observational study
found that higher plasma vitamin B6 levels were associated with better
performance on two measures of memory, but plasma vitamin B6 levels were
unrelated to performance on 18 other cognitive tests . Similarly, a double-
blind, placebo-controlled study in 38 healthy elderly men found that vitamin
B6 supplementation improved memory but had no effect on mood or mental
performance (19). Further, a placebo-controlled trial in 211 healthy younger,
middle-aged, and older women found that vitamin B6 supplementation (75
mg/day) for five weeks improved memory performance in some age groups but
had no effect on mood (20). Recently, a systematic review of randomized trials
concluded that there is inadequate evidence that supplementation with vitamin
B6, vitamin B12, or folic acid improves cognition in those with normal or
impaired cognitive function . Because of mixed findings, it is presently unclear
whether supplementation with B vitamins might blunt cognitive decline in the
elderly. Further, it is not known if marginal B vitamin deficiencies, which are
relatively common in the elderly, even contribute to age-associated declines in
cognitive function, or whether both result from processes associated with
aging and/or disease.
Kidney stones
A large prospective study examined the relationship between vitamin B6 intake
and the occurrence of symptomatic kidney stones in women. A group of more
than 85,000 women without a prior history of kidney stones were followed
over 14 years and those who consumed 40 mg or more of vitamin B6 daily had
only two thirds the risk of developing kidney stones compared with those who
consumed 3 mg or less . However, in a group of more than 45,000 men
followed over six years, no association was found between vitamin B6 intake
and the occurrence of kidney stones . Limited data have shown that
supplementation of vitamin B6 at levels higher than the tolerable upper intake
level (100 mg/day) decreases elevated urinary oxalate levels, an important
determinant of calcium oxalate kidney stone formation in some individuals.
However, it is less clear that supplementation actually resulted in decreased
formation of calcium oxalate kidney stones. Presently, the relationship
between vitamin B6 intake and the risk of developing kidney stones requires
further study before any recommendations can be made.
Disease Treatment
Vitamin B6 supplements at pharmacologic doses (i.e., doses much larger than
those needed to prevent deficiency) have been used in an attempt to treat a
wide variety of conditions, some of which are discussed below. In general, well
designed, placebo-controlled studies have shown little evidence that large
supplemental doses of vitamin B6 are beneficial .
Side effects of oral contraceptives
Because vitamin B6 is required for the metabolism of the amino acid
tryptophan, the tryptophan load test (an assay of tryptophan metabolites after
an oral dose of tryptophan) was used as a functional assessment of vitamin
B6 status. Abnormal tryptophan load tests in women taking high-dose oral
contraceptives in the 1960s and 1970s suggested that these women were
vitamin B6deficient. Abnormal results in the tryptophan load test led a number
of clinicians to prescribe high doses (100-150 mg/day) of vitamin B6 to women
in order to relieve depression and other side effects sometimes experienced
with oral contraceptives. However, most other indices of vitamin B6 status
were normal in women on high-dose oral contraceptives, and it is unlikely that
the abnormality in tryptophan metabolism was due to vitamin B6deficiency . A
more recent placebo-controlled study in women on the lower dose oral
contraceptives, which are commonly prescribed today, found that doses up to
150 mg/day of vitamin B6(pyridoxine) had no benefit in preventing side effects,
such as nausea, vomiting, dizziness, depression, and irritability .
Premenstrual syndrome (PMS)
The use of vitamin B6 to relieve the side effects of high-dose oral contraceptives
led to the use of vitamin B6 in the treatment of premenstrual syndrome (PMS).
PMS refers to a cluster of symptoms, including but not limited to fatigue,
irritability, moodiness/depression, fluid retention, and breast tenderness, that
begin sometime after ovulation (mid-cycle) and subside with the onset of
menstruation (the monthly period). A review of 12 placebo-controlled double-
blind trials on vitamin B6 use for PMS treatment concluded that evidence for a
beneficial effect was weak. A more recent review of 25 studies suggested that
supplemental vitamin B6, up to 100 mg/day, may be of value to treat PMS;
however, only limited conclusions could be drawn because most of the studies
were of poor quality .
Depression
Because a key enzyme in the synthesis of the neurotransmitters serotonin and
norepinephrine is PLP-dependent, it has been suggested that vitamin
B6 deficiency may lead to depression. However, clinical trials have not
provided convincing evidence that vitamin B6 supplementation is an effective
treatment for depression , though vitamin B6 may have therapeutic efficacy in
premenopausal women .
Morning sickness (nausea and vomiting in pregnancy)
Vitamin B6 has been used since the 1940s to treat nausea during pregnancy.
Vitamin B6 was included in the medication Bendectin, which was prescribed
for the treatment of morning sickness and later withdrawn from the market
due to unproven concerns that it increased the risk of birth defects. Vitamin
B6 itself is considered safe during pregnancy and has been used in pregnant
women without any evidence of fetal harm . The results of two double-blind,
placebo-controlled trials that used 25 mg of pyridoxine every eight hours for
three days or 10 mg of pyridoxine every eight hours for five days suggest that
vitamin B6 may be beneficial in alleviating morning sickness. Each study found
a slight but significant reduction in nausea or vomiting in pregnant women. A
recent systematic review of placebo-controlled trials on nausea during early
pregnancy found vitamin B6 to be somewhat effective. However, it should be
noted that morning sickness also resolves without any treatment, making it
difficult to perform well-controlled trials.
Carpal tunnel syndrome
Carpal tunnel syndrome causes numbness, pain, and weakness of the hand and
fingers due to compression of the median nerve at the wrist. It may result from
repetitive stress injury of the wrist or from soft tissue swelling, which
sometimes occurs with pregnancy or hypothyroidism. Several early studies by
the same investigator suggested that vitamin B6 status was low in individuals
with carpal tunnel syndrome and that supplementation with 100-200 mg/day
over several months was beneficial . A recent study in men not taking vitamin
supplements found that decreased blood levels of PLP were associated with
increased pain, tingling, and nocturnal wakening, all symptoms of carpal
tunnel syndrome . Studies using electrophysiological measurements of median
nerve conduction have largely failed to find an association between vitamin
B6deficiency and carpal tunnel syndrome. While a few trials have noted some
symptomatic relief with vitamin B6 supplementation, double-blind, placebo-
controlled trials have not generally found vitamin B6 to be effective in treating
carpal tunnel syndrome .
Sources
Food sources
Surveys in the U.S. have shown that dietary intake of vitamin B6averages about
2 mg/day for men and 1.5 mg/day for women. A survey of elderly individuals
found that men and women over 60 years old consumed about 1.2 mg/day and
1.0 mg/day, respectively; both intakes are lower than the current RDA. Certain
plant foods contain a unique form of vitamin B6 called pyridoxine glucoside;
this form of vitamin B6 appears to be only about half as bioavailable as vitamin
B6 from other food sources or supplements. Vitamin B6 in a mixed diet has
been found to be approximately 75% bioavailable . In most cases, including
foods in the diet that are rich in vitamin B6 should supply enough to prevent
deficiency. However, those who follow a very restricted vegetarian diet might
need to increase their vitamin B6 intake by eating foods fortified with vitamin
B6 or by taking a supplement. Some foods that are relatively rich in vitamin
B6 and their vitamin B6 content in milligrams (mg) are listed in the table below.
For more information on the nutrient content of specific foods, search the
USDA food composition database.
Food Serving Vitamin B6 (mg)
Fortified cereal 1 cup 0.5-2.5
Banana 1 medium 0.43
Salmon, wild, cooked 3 ounces* 0.48

Turkey, without skin,
3 ounces 0.39
cooked
Chicken, light meat

3 ounces 0.51
without skin, cooked
Potato, Russet, baked,

1 medium 0.70
with skin
Spinach, cooked 1 cup 0.44
Hazelnuts, dry roasted 1 ounce 0.18
Vegetable juice cocktail 6 ounces 0.26
*A 3-ounce serving of meat or fish is about the size of a deck of cards.

Supplements
Vitamin B6 is available as pyridoxine hydrochloride in multivitamin, vitamin B-
complex, and vitamin B6 supplements .
Safety
Toxicity
Because adverse effects have only been documented from vitamin
B6 supplements and never from food sources, safety concerning only the
supplemental form of vitamin B6 (pyridoxine) is discussed. Although vitamin
B6 is a water-soluble vitamin and is excreted in the urine, long-term
supplementation with very high doses of pyridoxine may result in painful
neurological symptoms known as sensory neuropathy. Symptoms include pain
and numbness of the extremities and in severe cases, difficulty walking.
Sensory neuropathy typically develops at doses of pyridoxine in excess of 1,000
mg per day. However, there have been a few case reports of individuals who
developed sensory neuropathies at doses of less than 500 mg daily over a
period of months. Yet, none of the studies in which an objective neurological
examination was performed reported evidence of sensory nerve damage at
intakes below 200 mg pyridoxine daily . To prevent sensory neuropathy in
virtually all individuals, the Food and Nutrition Board of the Institute of
Medicine set the tolerable upper intake level (UL) for pyridoxine at 100 mg/day
for adults (see table below) . Because placebo-controlled studies have generally
failed to show therapeutic benefits of high doses of pyridoxine, there is little
reason to exceed the UL of 100 mg/day.
Tolerable Upper Intake Level (UL) for Vitamin B6
Age Group UL (mg/day)
Infants 0-12 months Not possible to establish*
Children 1-3 years 30

Adolescents 14-18 years 80
Adults 19 years and older 100
*Source of intake should be from food and formula only.

Drug interactions
Certain medications interfere with the metabolism of vitamin B6; therefore,
some individuals may be vulnerable to a vitamin B6deficiency if supplemental
vitamin B6 is not taken. Anti-tuberculosis medications, including isoniazid and
cycloserine, the metal chelator penicillamine, and antiparkinsonian drugs
including L-dopa, all form complexes with vitamin B6 and thus create a
functional deficiency. Additionally, the efficacy of other medications may be
altered by high doses of vitamin B6. For instance, high doses of vitamin B6 have
been found to decrease the efficacy of two anticonvulsants, phenobarbital and
phenytoin, as well as L-dopa .
Linus Pauling Institute Recommendation
Metabolic studies suggest that young women require 0.02 mg of vitamin B6 per
gram of protein consumed daily . Using the upper boundary for acceptable
levels of protein intake for women (100 grams/day), the daily vitamin
B6 requirement for young women would be calculated at 2.0 mg daily. Older
adults may also require at least 2.0 mg/day. For these reasons, the Linus
Pauling Institute recommends that all adults consume at least 2.0 mg of
vitamin B6daily. Following the Linus Pauling Institute recommendation to take
a daily multivitamin-mineral supplement containing 100% of the Daily Value
for vitamin B6 will ensure an intake of at least 2.0 mg/day of vitamin
B6. Although a vitamin B6 intake of 2.0 mg daily is slightly higher than the most
recent RDA, it is 50 times less than the tolerable upper intake level (UL) set by
the Food and Nutrition Board (see Safety).
Older adults (65 years and older)
Metabolic studies have indicated that the requirement for vitamin B6 in older
adults is approximately 2.0 mg daily ; this requirement could be even higher if
the effect of marginally deficient vitamin B6intakes on immune function and
homocysteine levels are clarified. Despite evidence that the requirement for
vitamin B6 may be slightly higher in older adults, several surveys have found
that over half of individuals over age 60 consume less than the current RDA
(1.7 mg/day for men and 1.5 mg/day for women). For these reasons, the Linus
Pauling Institute recommends that older adults take a
multivitamin/multimineral supplement, which generally provides at least 2.0
mg of vitamin B6 daily.

Labels: dopamine synthesis, GABA synthesis, norepinephrine synthesis,pyridoxal
phosphate, pyridoxamine, pyridoxine, serotonin synthesis, tryptophan load test, vitamin
b6, xanthinuric acid
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Biochemistry

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43 - Ninhydrin test or Ninhydrin reagent

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A ninhydrin solution is commonly used by forensic investigators in the analysis of latent

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Increasing the sensitivity of the biuret test :

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A reddish-brown coloration or precipitate indicates the presence of tyrosine residues which

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*THB is biosynthesized from guanosine triphosphate (GTP) by three chemical reactions,

*THB has the following responsibilities as a cofactor:

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36 - Diseases caused by mutations in collagen genes

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35 - AIIMS MAY 2001 biochemistry mcqs with answers

2q) Amber codon refers to

3q) At physiological pH, the most stable amino

4q) In cystinuria, amino acids excreted are all the

5q) Dietary triglycerides are transported by

6q) In which of the following reaction, thiamine

a) Alpha ketoglutarate to succinyl CoA

7q) In chromatography, mass movement of the

8q) The type of chromatography in which

9q) The end-product of citric acid cycle used in

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34 - AIIMS november 2002 biochemistry mcqs

explanation : alfa helix of a protein can be disrupted due to the introduction of

2. charged aminoacids : glutamate ,aspartate, lysine, arginine and histidine .

3. aminoacids with bulky side chains : tryptophan

4. aminoacids that branch at the beta-carbon : valine and isoleucine .

1q: dna without introns is?

2q: all are true about glutathione except?

A. Converts hemoglobin to methemoglobin

3q: Ribosome has followin enzymatic activity?

4q: which enzyme is responsible for carboxylation reaction?

5q: Glowing of firefly is due to?

6q: in carboxylation of clotting factors by vit k which amino acid is

7q: synthesis of a immunoglobulin in membrane bound or independent form is

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8q: phosphorlase b is inhibited by?

9q: In metabolism of xenobiotics all of the followin reactions occour in phase

10q: Which is not a second messenger?

11q: structure of proteins can be determined by all except?

12q: Functions of thiamine?

A. Co enzyme of pyruvate dehydrogenase and alpha keto dehydrogenase

WEDNESDAY, NOVEMBER 12, 2008

32 - AIIMS novemeber 2008 biochemistry mcqs - 1

The answer is ATP….

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Coenorhabditis elegans (a nematode) was the FIRST MULTICELLULAR ORGANISM

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To study the transcriptional activity of a gene , incorporate the luciferase gene

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MONDAY, AUGUST 18, 2008

31 - Rate limiting enzymes of biochemical pathways