Professional Documents
Culture Documents
Detoxification
27/9-9/10/2011
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References:
1. "Biochemistry" by Lubert Stryer
(textbook)
Prof. Dr.H.D.El-Yassin1
2011
LECTURE 1
Tuesday 27/9/2011
maltose
proteins
peptides
fats
glucose
amino acids
Prof. Dr.H.D.El-Yassin2
2011
LECTURE 1
Tuesday 27/9/2011
LECTURE 1
Tuesday 27/9/2011
Prof. Dr.H.D.El-Yassin3
2011
Carbohydrates
1. Monosaccharides or simple sugars are either hexoses (6-carbon) like
glucose, galactose and fructose, or pentoses (5-carbon) like ribose.
These are the breakdown products of more complex carbohydrates and
can be efficiently absorbed across the wall of the digestive tube and
transported into blood.
2. Disaccharides are simply two monosaccharides linked together by a
glycosidic bond. The disaccharides most important in nutrition and
digestion are:
3. Polysaccharides :
LECTURE 1
Tuesday 27/9/2011
Prof. Dr.H.D.El-Yassin4
2011
The process of digestion produces glucose, amino acids, glycerol, and fatty
acids (see above). The energy in glucose is used to produce ATP via the
reactions of glycolysis, cellular respiration, and the electron transport system
(see diagram below). The body uses amino acids to construct proteins.
Excess amino acids can be used to synthesize pyruvate, acetyl CoA, and
alpha ketogluterate, which enters the Krebs cycle. Glycerol and fatty acids
can be converted to pyruvate and Acetyl CoA and then enter cellular
respiration.
Mouth
Chewing breaks food into smaller particles so that chemical digestion can
occur faster.
Enzymes: Salivary amylase breaks starch (a polysaccharide) down to
maltose (a disaccharide).
Bicarbonate ions in saliva act as buffers, maintaining a pH between 6.5
and 7.5.
Mucins (mucous) lubricate and help hold chewed food together in a
clump called a bolus.
Prof. Dr.H.D.El-Yassin5
2011
LECTURE 1
Tuesday 27/9/2011
Stomach
The stomach stores up to 2 liters of food. Gastric glands within the stomach
produce secretions called gastric juice.
The muscular walls of the stomach contract vigorously to mix food with gastric
juice, producing a mixture called chyme.
Gastric juice
Pepsinogen is converted to pepsin, which digests proteins. Pepsinogen
production is stimulated by the presence of gastrin in the blood.
HCl
Hydrochloric acid (HCl) converts pepsinogen to pepsin which breaks down
proteins to peptides. HCl maintains a pH in the stomach of approximately 2.0.
It also dissolves food and kills microorganisms.
Mucous protects the stomach from HCl and pepsin.
Secretion of Gastric Juice: Gastrin is a hormone that stimulates the
stomach to secrete gastric juice.
Duodenum
The duodenum is the first part of the small intestine.
Chyme enters in tiny spurts. At this point, proteins and carbohydrates are only
partially digested and lipid digestion has not begun.
Pancreas
The pancreas acts as an exocrine gland by producing pancreatic juice which
empties into the small intestine via a duct.
The pancreas also acts as an endocrine gland to produce insulin.
Pancreatic Juice
Pancreatic juice contains sodium bicarbonate which neutralizes the acidic
material from the stomach.
Pancreatic amylase digests starch to maltose.
Trypsin and Chymotrypsin digest proteins to peptides. Like pepsin
(produced in the stomach), they are specific for certain amino acids, not
all of them. They therefore produce peptides.
Lipase digests fats to glycerol and fatty acids.
Prof. Dr.H.D.El-Yassin6
2011
LECTURE 1
Tuesday 27/9/2011
Liver
The liver produces bile which is stored in gallbladder and sent to the
duodenum through a duct.
Bile emulsifies fats (separates it into small droplets) so they can mix with
water and be acted upon by enzymes.
Other Functions of the Liver
The liver detoxifies blood from intestines that it receives via the hepatic
portal vein.
The liver stores glucose as glycogen (animal starch) and breaks down
glycogen to release glucose as needed. This storage-release process
maintains a constant glucose concentration in the blood (0.1%). If
glycogen and glucose run short, proteins can be converted to glucose.
It produces blood proteins.
It destroys old red blood cells and converts hemoglobin from these cells
to bilirubin and biliverdin which are components of bile.
Ammonia produced by the digestion of proteins is converted to a less
toxic compound (urea) by the liver.
Prof. Dr.H.D.El-Yassin7
2011
It has the opposite effects of gastrin; it inhibits gastric glands in the stomach
and it inhibits the mixing and churning movement of stomach muscles. This
slows the rate of stomach emptying when the duodenum contains food.
LECTURE 1
Tuesday 27/9/2011
Small Intestine
The small intestine is approximately 3 m long. Like the stomach, it contains
numerous ridges and furrows. In addition, there are numerous projections
called villi that function to increase the surface area of the intestine. Individual
villus cells have microvilli which greatly increase absorptive surface area.
The total absorptive surface area is equivalent to 500 or 600 square meters.
Each villus contains blood vessels and a lacteal (lymph vessel).
Peptidases and maltase are embedded within the plasma membrane of the
microvilli.
Peptidases complete the digestion of peptides to amino acids.
Maltase completes the digestion of disaccharides.
Absorption:
Prof. Dr.H.D.El-Yassin8
2011
LECTURE 1
Tuesday 27/9/2011
ENZYME
SOURCE
PRODUCTS
CARBOHYDRATES
Salivary amylase
Pancreatic amylase
Maltase
Salivary glands
Pancreas
Small intestine
Maltose
Maltose
Glucose
PROTEINS
Pepsin
Trypsin
Peptidases
Stomach mucosa
Pancreas
Intestinal mucosa
Peptides
Peptides
Amino acids
FATS
Lipase
Pancreas
Fatty acids
and glycerol
ENZYME
FOOD
PRODUCT
MOUTH (salivary
glands)
Salivary amylase
Polysaccharides
Maltose
STOMACH
Pepsin
Proteins
Peptides
PANCREAS
Pancreatic
amylase
Trypsin
Lipase
Polysaccharides
Proteins
Fats
Maltose
Peptides
Fatty acids
and glycerol
SMALL INTESTINE
Maltase
Peptidases
Maltose
Peptides
Glucose
Amino acids
Prof. Dr.H.D.El-Yassin9
2011
LECTURE 2
Thursday 29/9/2011
The Stomach
Foodstuffs entering the stomach have been, crushed and reduced
in size by mastication, with saliva. The stomach provides four basic
functions that assist in the early stages of digestion and prepare the
ingesta for further processing in the small intestine:
1. It serves as a short-term storage reservoir, allowing a rather
large meal to be consumed quickly and dealt with over an extended
period.
2. It is in the stomach that substantial chemical and enzymatic
digestion is initiated, particularly of proteins.
3. Vigorous contractions of gastric smooth muscle mix and grind
foodstuffs with gastric secretions, resulting in liquefaction of
food, a prerequisite for delivery of the ingesta to the small
intestine.
4. As food is liquefied in the stomach, it is slowly released into
the small intestine for further processing.
Prof. Dr.H.D.El-Yassin
10
2011
If the lining of the stomach is examined with a hand lens, one can
see that it is covered with numerous small holes. These are the
openings of gastric pits which extend into the mucosa as straight and
branched tubules, forming gastric glands.
LECTURE 2
Thursday 29/9/2011
Four major types of secretory epithelial cells cover the surface of the
stomach and extend down into gastric pits and glands:
LECTURE 2
Thursday 29/9/2011
Gastric secretions
1. Mucosal Protection
Mucus layer on gastric surface forms a mucosal barrier to damage
against several forms of potential injury to the gastric mucosa.
1. A gel 0.2mm thick; 80% CHO; 20% protein
2. Secreted by neck cells, surface epithelium
3. Can be cleaved by pepsin, so continual production is required
4. Release is stimulated by acetylcholine from nerve endings
5. Also rich in bicarbonate
a. HCO3- content creates a "micro-environment" around surface
cells to prevent acid damage
b. HCO3- secretion is inhibited by adrenergic input (prominent in
stress)
2. Acid Secretion
Hydrochloric acid is secreted from parietal cells into the lumen where
it establishes an extremely acidic environment. This acid is important
for activation of pepsinogen and inactivation of ingested
microorganisms such as bacteria.
2.1. Function of Gastric acid
1.
LECTURE 2
Thursday 29/9/2011
Prof. Dr.H.D.El-Yassin
13
2011
LECTURE 2
Thursday 29/9/2011
Prof. Dr.H.D.El-Yassin
14
2011
Hydrogen ion is pumped out of the cell, into the lumen, in exchange
for potassium through the action of the proton pump; potassium is thus
effectively recycled.
Accumulation of osmotically-active hydrogen ion in the cannaliculus
generates an osmotic gradient across the membrane that results in
outward diffusion of water - the resulting gastric juice is 155 mM
HC1 and 15 mM KC1 with a small amount of NaCl.
LECTURE 2
Thursday 29/9/2011
GASTRIN
o binds to CCK-B receptor on cell surface
o releases intracellular Ca++
HISTAMINE
o released from mast cells
o binds to parietal cell surface receptor
o activates adenyl cyclase (increases cyclic AMP, an
intracellular messenger)
Prof. Dr.H.D.El-Yassin
15
2011
LECTURE 2
Thursday 29/9/2011
HCO3
H 2 CO3
Lactate
Lactic acid
All tend to increase on the side of the base i.e.:HPO4-2, HCO3and lactate, with the result that the pH of plasma is raised and an
alkaline urine is excreted for some hours following intake of food
and gastric secretion. This is known as the alkaline tide.
Prof. Dr.H.D.El-Yassin
17
2011
LECTURE 2
Thursday 29/9/2011
3. Proteases:
Pepsinogen, an inactive zymogen, is secreted into gastric juice from
both mucous cells and chief cells. Once secreted, pepsinogen is
activated by stomach acid into the active protease pepsin, which is
largely responsible for the stomach's ability to initiate digestion of
proteins, in young animals; chief cells also secrete chymosin (rennin),
a protease that coagulates milk protein allowing it to be retained more
than briefly in the stomach.
Pepsinogens and Pepsins
Pepsinogens are secreted in a form such that the activation
peptide assumes a compact structure that occludes the active site.
On exposure to an acidic (pH < 4) environment such as occurs in the
lumen of the stomach, the activation peptide unfolds, allowing the
active site to clip it off, yielding mature, catalytically active pepsin.
Optimal activity of pepsins is at pH of 1.8 to 3.5, depending on the
isoform, They are reversibly inactivated at about pH 5 and irreversibly
inactivated at pH 7 to 8.
The mature, active enzymes are roughly 325 amino acids with a
mass of approximately 35 kDa.
Pepsin initiates protein digestion by splitting certain amino acid linkages
in proteins (Cleaves preferentially C-terminal. It does not cleave at V, A or
G. Other residues may be cleaved, with very variable rates) to yield
peptide fragments.
LECTURE 2
Thursday 29/9/2011
LECTURE 2
Thursday 29/9/2011
4. Hormones
The principle hormone secreted from the gastric epithelium is gastrin,
a peptide that is important in control of acid secretion and gastric
motility. Gastrin is secreted by G-cells and released into the blood
where it travels to the parietal cells to stimulate acid secretion, and to
Enterochromaffin-Like (ECL) Cells to stimulate histamine secretion.
The net result of gastrin secretion is increased acid production
through two mechanisms:
1. Direct stimulation of the parietal cells,
2. Tropic action on parietal cells increasing their number.
N.B. in gastrinoma (Zollinger-Ellison syndrome) increased production
of gastrin causes hypersecretion of acid which is not subject to normal
inhibitory mechanisms.
A number of other enzymes are secreted by gastric epithelial cells,
including a lipase and gelatinase. One secretory product of
considerable importance in man is intrinsic factor, a glycoprotein
secreted by parietal cells that is necessary for intestinal absorption of
vitamin B12.
LECTURE 2
Thursday 29/9/2011
Intrinsic Factor
Intrinsic factor is a glycoprotein secreted by parietal (humans) of the
gastric mucosa. In humans, it has an important role in the absorption of
vitamin B12 (cobalamin) in the intestine, and failure to produce or
utilize intrinsic factor results in the condition pernicious anemia.
as a result of an autoimmune attack against parietal cells
Dietary vitamin B12 is released from ingested proteins in the
stomach through the action of pepsin and acid. It is rapidly bound
by one of two vitamin B12-binding proteins that are present in gastric
juice; at acid pH, these binding proteins have a greater affinity for
the vitamin than does intrinsic factor. In the small intestine
pancreatic proteases digest the binding proteins, releasing vitamin
B12 which then becomes bound to intrinsic factor. Finally, there are
receptors for intrinsic factor on the ileal mucosa which bind the
complex, allowing vitamin B12 to be absorbed into portal blood.
In all mammals, vitamin B12is necessary for maturation of erythrocytes,
and a deficiency of this vitamin leads to development of anemia.
Since efficient absorption of vitamin B12in humans depends on
intrinsic factor, diseases which decrease the secretion of intrinsic
factor (e.g. atrophic gastritis), interfere with cleavage of the binding
proteins (e.g. pancreatic exocrine insufficiency) or decrease binding
and absorption of the intrinsic factor-vitamin B12 complex (e.g. ileal
disease or resection) can result in this type of anemia.
Absorption in the Stomach
The stomach absorbs very few substances, although small amounts
of certain lipid-soluble compounds can be taken up, including
aspirin, other non-steroidal anti-inflammatory drugs, and ethanol.
Notably, these substances are also well-recognized causes of
gastric irritation and their use (especially overuse) is commonly
associated with development of gastritis and gastric ulcers.
LECTURE 3
Sunday 2/10/2011
The Pancreas
The pancreas plays a vital role in accomplishing the followings:
Acid must be quickly and efficiently neutralized to prevent damage to the
duodenal mucosa
Macromolecular nutrients - proteins, fats and starch - must be broken down
much further before their constituents can be absorbed through the mucosa into
blood
Insufficient exocrine secretion by the pancreas leads to starvation, even if the body is
consuming adequate quantities of high quality food.
In addition to its role as an exocrine organ, the pancreas is also an endocrine organ.
The major hormones it secretes - insulin and glucagon - play a vital role in
carbohydrate and lipid metabolism.
LECTURE 3
Sunday 2/10/2011
pancreatic
proteases
are
trypsin
and
chymotrypsin
both
are
LECTURE 3
Sunday 2/10/2011
Once trypsinogen and chymotrypsinogen are released into the lumen of the small
intestine, they must be converted into their active forms in order to digest proteins,
Trypsinogen is activated by the enzyme enterokinase, which is embedded in the
intestinal mucosa.
Once trypsin is formed, it activates chymotrypsinogen, as well as additional molecules
of trypsinogen. The net result is a rather explosive appearance of active protease
once the pancreatic secretions reach the small intestine.
Trypsin and chymotrypsin digest proteins into peptides and peptides into smaller
peptides, but they cannot digest proteins and peptides to single amino acids. Some
of the other proteases from the pancreas, for instance carboxypeptidase
(exopeptidase) (This enzyme removes, one by one, the amino acids at the C-terminal of peptides).
But the final digestion of peptides into amino acids is largely the effect of peptidases in small intestinal
epithelial cells.
LECTURE 3
Sunday 2/10/2011
b. Pancreatic Lipase
The major form of dietary fat is triglyceride, or neutral lipid. A triglyceride molecule
cannot be directly absorbed across the intestinal mucosa. It must first be digested
into a 2-monoglyceride and two free fatty acids. The enzyme that performs
this hydrolysis is pancreatic lipase.
Sufficient quantities of bile salts must also be present in the lumen of the intestine
in order for lipase to efficiently digest dietary triglyceride and for the resulting
fatty acids and monoglyceride to be absorbed. This means that normal
digestion and absorption of dietary fat is critically dependent on secretions from
both the pancreas and liver.
Pancreatic lipase has recently been in the limelight as a target for management
of obesity. The drug orlistat (Xenical) is a pancreatic lipase inhibitor that
interferes with digestion of triglyceride and thereby reduces absorption of
dietary fat. Clinical trials support the contention that inhibiting lipase can lead to
significant reductions in body weight in some patients.
c. Amylase
The major dietary carbohydrate for many species is starch, a storage form of glucose in
plants. Amylase is the enzyme that hydrolyses starch to maltose (a glucose-glucose
disaccharide), as well as the trisaccharide maltotriose and small branchpoints fragments
called dextrins.
d. Other Pancreatic Enzymes
In addition to the proteases, lipase and amylase, the pancreas produces a host of
other digestive enzymes, including nucleases, gelatinase and elastase.
Nucleases. These hydrolyze ingested nucleic acids (RNA and DNA) into their
component nucleotides.
Elastase: Cuts peptide bonds next to small, uncharged side chains such as
those of alanine and serine.
LECTURE 3
Sunday 2/10/2011
LECTURE 3
Sunday 2/10/2011
The most important stimuli for pancreatic secretion come from three hormones secreted
by the enteric endocrine system:
Cholecystokinin: This hormone is synthesized and secreted by enteric endocrine
cells located in the duodenum. Its secretion is strongly stimulated by the presence of
partially digested proteins and fats in the small intestine. As chyme floods into the
small intestine, cholecystokinin is released into blood and binds to receptors on
pancreatic acinar cells, ordering them to secrete large quantities of digestive
enzymes. It also stimulates the gallbladder to release bile and the pancreas to
produce pancreatic enzymes.
LECTURE 3
Sunday 2/10/2011
The Liver
The liver is the largest gland in the body and performs an astonishingly large
number of tasks that impact all body systems. One consequence of this complexity is
that hepatic disease has widespread effects on virtually all other organ systems.
The three fundamental roles of the liver are:
1. Vascular functions: including formation of lymph and hepatic phagocytic system.
2. Metabolic achievements in control of synthesis and utilization of
carbohydrates, lipids and proteins.
3. Secretory and excretory functions, particularly with respect to the synthesis
of secretion of bile.
The latter is the only one of the three that directly affects digestion - the liver,
through its biliary tract, secretes bile acids into the small intestine where they
assume a critical role in the digestion and absorption of dietary lipids.
LECTURE 3
Sunday 2/10/2011
LECTURE 3
Sunday 2/10/2011
1. Cholic acid
2. Deoxycholic acid
3. Decholin
4. Chenodiol (Chenix), is used to dissolve gallstones in patients who cannot
tolerate surgery. Chenodiol is a natural bile acid that blocks production of
cholesterol . This action leads to gradual dissolution of cholesterol gallstones.
LECTURE 3
Sunday 2/10/2011
Synthesis of bile acids is one of the predominant mechanisms for the excretion of
excess cholesterol. However, the excretion of cholesterol in the form of bile acids is
insufficient to compensate for an excess dietary intake of cholesterol.
These acids are then conjugated with glycine or taurine and secreted as Na + (or K+)
salts. Conjugation causes a decrease in their pKa values, making them more water
soluble.
The most abundant bile acids in human bile are chenodeoxycholic acid (45%) and
cholic acid (31%). These are referred to as the primary bile acids. Within the
intestines the primary bile acids are acted upon by bacteria and converted to the
secondary bile acids, identified as deoxycholate (from cholate) and lithocholate
(from chenodeoxycholate). Both primary and secondary bile acids are reabsorbed by
the intestines and delivered back to the liver via the portal circulation. Within the
liver the carboxyl group of primary and secondary bile acids is conjugated via an
amide bond to either glycine or taurine before their being resecreted into the bile
canaliculi.
These conjugation reactions yield glycoconjugates and tauroconjugates, respectively.
The bile canaliculi join with the bile ductless, which then form the bile ducts. Bile acids
are carried from the liver through these ducts to the gallbladder, where they are stored
for future use. The ultimate fate of bile acids is secretion into the intestine, where they
aid in the emulsification of dietary lipids. In the gut the glycine and taurine residues are
removed and the bile acids are either excreted (only a small percentage) or
reabsorbed by the gut and returned to the liver. This process of secretion from the liver
to the gallbladder, to the intestines and finally reabsorbtion is termed the enterohepatic
circulation.
LECTURE 3
Sunday 2/10/2011
Enterohepatic Recirculation
After the bile acids has been released into the small intestine via the bile duct to play an
integral role in the absorption of dietary lipids and lipid soluble vitamins. More than
90% of the bile salts are actively reabsorbed (by a sodium-dependent cotransport process) from the ileum into the hepatic-portal circulation from where they are
cleared and resecreted by the liver to once again be stored in the gall bladder. This
secretion/reabsorption cycle is called the Enterohepatic Circulation.
Systemic circulation: supplies nourishment to all of the tissue located throughout your
body, with the exception of the heart and lungs because they have their own systems.
Systemic circulation is a major part of the overall circulatory system.
Portal circulation: Blood from the gut and spleen flow to and through the liver before
returning to the right side of the heart. This is called the portal circulation and the large vein
through which blood is brought to the liver is called the portal vein.
The net effect of this enterohepatic recirculation is that each bile salt molecule is
reused about 20 times, often two or three times during a single digestive phase.
LECTURE 3
Sunday 2/10/2011
Note: liver disease can dramatically alter this pattern of recirculation - for instance, sick
hepatocytes have decreased ability to extract bile acids from portal blood and damage to the
canalicular system can result in escape of bile acids into the systemic circulation. Assay of
systemic levels of bile acids is used clinically as a sensitive indicator of hepatic disease.
Bile acids are facial amphipathic, that is, they contain both hydrophobic
(lipid soluble) and polar (hydrophilic) faces. The cholesterol-derived portion of
a bile acid has one face that is hydrophobic (that with methyl groups) and one
that is hydrophilic (that with the hydroxyl groups); the amino acid conjugate is
polar and hydrophilic.
Their amphipathic nature enables bile acids to carry out two important
functions:
1. Emulsification of lipid aggregates: Bile acids have detergent action
on particles of dietary fat, which causes fat globules to break down
or be emulsified into minute, microscopic droplets. Emulsification is not
digestion per se, but is of importance because it greatly increases the
surface area of fat, making it available for digestion by lipases, which
cannot access the inside of lipid droplets.
2. Solubilization and transport of lipids in an aqueous environment: Bile
acids are lipid carriers and are able to solubilize many lipids by
forming micelles - aggregates of lipids such as fatty acids, cholesterol
and monoglycerides - that remain suspended in water. Bile acids are
also critical for transport and absorption of the fat-soluble vitamins.
34
Prof.Dr H.D.El-Yassin
2011
LECTURE 3
Sunday 2/10/2011
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Prof.Dr H.D.El-Yassin
2011
LECTURE 3
Sunday 2/10/2011
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
The liver plays several roles in detoxification: it filters the blood to remove
large toxins, synthesizes and secretes bile full of cholesterol and other fatsoluble toxins, and enzymatically disassembles unwanted chemicals.
LECTURE 4
Tuesday 4/10/2011
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
phase I and
phase II.
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
Phase I Detoxification
This pathway converts a toxic chemical into a less harmful chemical. This is
achieved by various chemical reactions (such as oxidation, reduction and
hydrolysis), and during this process free radicals are produced which, if
excessive, can damage the liver cells. Antioxidants reduce the damage
caused by these free radicals. If antioxidants are lacking and toxin exposure
is high, toxic chemicals become far more dangerous. Some may be
converted from relatively harmless substances into potentially carcinogenic
substances.
The effects of exposure to toxins varies from individual to individual. Some
people are highly sensitive to different endogenous and exogenous toxins.
Others, because their bodies are more resilient and their livers can detoxify
more efficiently, aren't as sensitive.
CYTOCHROME P450 MONOOXYGENASE SYSTEM
Monooxygenase (mixed function oxidases) incorporate one atom from
molecular oxygen into a substrate (creating a hydroxyl group), with the
other atom being reduced to water. In the cytochrome P450
monooxygenase system NADPH provides the reducing equivalents
required by the series of reactions. This system performs different
functions in two separate locations in cells.The overall reaction catalyzed
by a cytochrome P450 enzyme is:
R-H + O2 + NADPH + H+
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
Tuesday 4/10/2011
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
Phase II Detoxification
This is called the conjugation pathway, whereby the liver cells add
another substance (eg. cysteine, glycine or a sulphur molecule) to a toxic
chemical or drug. This makes the toxin or drug water-soluble, so it can then
be excreted from the body via watery fluids such as bile or urine. Individual
xenobiotics and metabolites usually follow one or two distinct pathways.
There are essentially six phase II detoxification pathways:
1. Glutathione conjugation
2. Amino acid conjugation
3. Methylation
4. Sulfation
5. Acetylation
6. Glucuronidation
1. Glutathione conjugation
A primary phase II detoxification route is
conjugation with glutathione(glutamylcysteinylglycine), (a
tripeptide composed of three amino acids-cysteine, glutamic acid, and glycine).
Glutathione conjugation produces watersoluble mercaptates which are excreted via
the kidneys.
The elimination of fat-soluble compounds,
especially heavy metals like mercury and lead, is dependent upon adequate
levels of glutathione, which in turn is dependent upon adequate levels of
methionine and cysteine. When increased levels of toxic compounds are
present, more methionine is utilized for cysteine and glutathione synthesis.
Methionine and cysteine have a protective effect on glutathione and prevent
depletion during toxic overload. This, in turn, protects the liver from the
damaging effects of toxic compounds and promotes their elimination.
If the availability of methionine is reduced, not only will the capability of
the liver to detoxify be impaired, but there will also be less glutathione
available to complex with foreign substances.
Studies have demonstrated that a deficiency of methionine can, in itself,
cause liver cancer without the presence of a carcinogen, and also that
the deficiency of methionine can permit a heavy metal to cause toxic
effects.
LECTURE 4
Tuesday 4/10/2011
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Prof.Dr H.D.El-Yassin
2011
LECTURE 4
Tuesday 4/10/2011
LECTURE 4
Tuesday 4/10/2011
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Prof.Dr H.D.El-Yassin
2011
4. Sulfation
Sulfation is the conjugation of toxins with sulfur-containing compounds. The
sulfation system is important for detoxifying several drugs, food additives,
and, especially, toxins from intestinal bacteria and the environment. In
addition to environmental toxins, sulfation is also used to detoxify some
normal body chemicals and is the main pathway for the elimination of
steroid and thyroid hormones. Since sulfation is also the primary route for
the elimination of neurotransmitters, dysfunction in this system may
contribute to the development of some nervous system disorders.
Many factors influence the activity of sulfate conjugation. For example, a
diet low in methionine and cysteine has been shown to reduce sulfation.
5. Acetylation
Conjugation of toxins with acetyl-CoA is the primary method by which the
body eliminates sulfa drugs. This system appears to be especially sensitive
to genetic variation, with those having a poor acetylation system being far
more susceptible to sulfa drugs and other antibiotics. While not much is
known about how to directly improve the activity of this system, it is known
that acetylation is dependent on thiamine, pantothenic acid, and vitamin C.
6. Glucuronidation
Glucuronidation, the combining of glucuronic acid with toxins, in Phase II
can be reversed by Beta glucuronidase enzymes produced by pathological
bacteria and cause toxins to be reabsorbed increasing toxicity. Many of the
commonly prescribed drugs are detoxified through this pathway. It also
helps to detoxify aspirin, menthol, vanillin (synthetic vanilla), food additives
such as benzoates, and some hormones.
Sulfoxidation
Sulfoxidation is the process by which the sulfur-containing molecules in
drugs and foods are metabolized. It is also the process by which the body
eliminates the sulfite food additives used to preserve many foods and
drugs. Normally, the enzyme sulfite oxidase (molybdenum
dependentenzyme) metabolizes sulfites to safer sulfates, which are then
excreted in the urine. Those with a poorly functioning sulfoxidation system,
however, have an increased ratio of sulfite to sulfate in their urine. Those
with a poorly functioning sulfoxidation detoxification pathway are more
sensitive to sulfur-containing drugs and foods containing sulfur or sulfite
additives.
Lecture 5
Lecture 5
Thursday 6/10/2011
Thursday 6/10/2011
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Prof.Dr H.D.El-Yassin
2011
Lecture 5
Thursday 6/10/2011
45
Prof.Dr H.D.El-Yassin
2011
Lecture
5
Thursday 6/10/2011
CLINICAL CORRELATION
Cystic fibrosis
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Abnormal activation of the cAMP-dependent chloride channel (CFTR) in crypt cells has
resulted in the deaths of millions upon millions of people. Several types of bacteria produce
toxins that strongly, often permanently, activate the adenylate cyclase in crypt enterocytes.
This leads to elevated levels of cAMP, causing the chloride channels to essentially become
stuck in the "open" position". The result is massive secretion of water that is manifest as
severe diarrhea. Cholera toxin, produced by cholera bacteria, is the best known example of
this phenomenon, but several other bacteria produce toxins that act similarly.
Absorption in the Small Intestine: General Mechanisms
Virtually all nutrients from the diet are absorbed into blood across the mucosa of the
small intestine.To remain viable, all cells are required to maintain a low intracellular
concentration of sodium. In polarized epithelial cells like enterocytes, low intracellular
sodium is maintained by a large number of Na +/K+ ATPases - so-called sodium
pumps - embedded in the basolateral membrane. These pumps export 3 sodium ions
from the cell in exchange for 2 potassium ions, thus establishing a gradient of both
charge and sodium concentration across the basolateral membrane.
Aside from the electrochemical gradient of sodium, several other concepts are
required to understand absorption in the small intestine. Also, dietary sources of
protein, carbohydrate and fat must all undergo the final stages of chemical digestion
just prior to absorption of, for example, amino acids, glucose and fatty acids.
Lecture 5
Thursday 6/10/2011
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2011
The small intestine must absorb massive quantities of water. A normal person akes in roughly
1 to 2 liters of dietary fluid every day. On top of that, another 6 to 7 liters of fluid is received
by the small intestine daily as secretions from salivary glands, stomach, pancreas, liver and
the small intestine itself.
By the time the ingesta enters the large intestine, approximately 80% of this fluid has been
absorbed. Net movement of water across cell membranes always occurs by osmosis, and the
fundamental concept needed to understand absorption in the small gut is that absorption of
water is absolutely dependent on absorption of solutes, particularly sodium:
Sodium is absorbed into the cell by several mechanisms, but chief among
them is by co-transport with glucose and amino acids - this means that
efficient sodium absorption is dependent on absorption of these organic
solutes.
Absorbed sodium is rapidly exported from the cell via sodium pumps - when a
lot of sodium is entering the cell, a lot of sodium is pumped out of the cell,
which establishes a high osmolarity in the small intercellular spaces between
adjacent enterocytes.
Water diffuses in response to the osmotic gradient established by sodium - in
this case into the intercellular space. It seems that the bulk of the water
absorption is transcellular, but some also diffuses through the tight junctions.
Water, as well as sodium, then diffuses into capillary blood within the villus.
Water is thus absorbed into the intercellular space by diffusion down an osmotic gradient.
However, looking at the process as a whole, transport of water from lumen to blood is often
against an osmotic gradient - this is important because it means that the intestine can absorb
water into blood even when the osmolarity in the lumen is higher than osmolarity of blood.
2) Absorption of Monosaccharides
Monosaccharides, are only rarely found in normal diets. Rather, they are derived by
enzymatic digestion of more complex carbohydrates within the digestive tube.
Particularly important dietary carbohydrates include starch and disaccharides such as lactose
and sucrose. None of these molecules can be absorbed for the simple reason that they cannot
cross cell membranes unaided and, unlike the situation for monosaccharides, there are no
transporters to carry them across.
Brush Border Hydrolases Generate Monosaccharides
Polysaccharides and disaccharides must be digested to monosaccharides prior to absorption
and the key players in these processes are the brush border hydrolases, which include maltase,
lactase and sucrase. Dietary lactose and sucrose are "ready" for digestion by their respective
brush border enzymes. Starch, as discussed previously, is first digested to maltose by amylase
in pancreatic secretions and, saliva.
Lecture 5
Thursday 6/10/2011
Dietary lactose and sucrose, and maltose derived from digestion of starch, diffuse in
the small intestinal lumen and come in contact with the surface of absorptive
epithelial cells covering the villi where they engage with brush border hydrolases:
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1. the transporter is initially oriented facing into the lumen - at this point it is capable of
binding sodium, but not glucose
2. sodium binds, inducing a conformational change that opens the glucose-binding
pocket
3. glucose binds and the transporter reorients in the membrane such that the pockets
holding sodium and glucose are moved inside the cell
4. sodium dissociates into the cytoplasm, causing glucose binding to destabilize
5. glucose dissociates into the cytoplasm and the unloaded transporter reorients back to
its original, outward-facing position
Thursday 6/10/2011
CLINICAL CORRELATION
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Disaccharidase Deficiency
Intestinal disaccharidase deficiencies are encountered relatively frequently in humans.
Deficiency can be present in one enzyme or several enzymes for a variety of reasons (genetic
defect, physiological decline with age, or the result of "injuries" to the mucosa). Of the
disaccharidases, lactase is the most common enzyme with an absolute or relative deficiency,
which is experienced as milk intolerance. The consequences of an inability to hydrolyze
lactose in the upper small intestine are inability to absorb lactose and bacterial fermentation of
ingested lactose in the lower small intestine. Bacterial fermentation results in the production
of gas (distension of gut and flatulence) and osmotically active solutes that draw water into
the intestinal lumen (diarrhea). The lactose in yogurt has already been partially hydrolyzed
during the fermentation process of making yogurt. Thus individuals with lactase deficiency
can often tolerate yogurt better than unfermented dairy products. The enzyme lactase is
commercially available to pretreat milk so that the lactose is hydrolyzed.
3) Absorption of Amino Acids and Peptides
Dietary proteins are, with very few exceptions, not absorbed. Rather, they must be
digested into amino acids or di- and tripeptides first, through the action of gastric and
pancreatic proteases. The brush border of the small intestine is equipped with a
family of peptidases. Like lactase and maltase, these peptidases are integral
membrane proteins rather than soluble enzymes. They function to further the
hydrolysis of lumenal peptides, converting them to free amino acids and very small
peptides. These end products of digestion, formed on the surface of the enterocyte,
are ready for absorption.
a) Absorption of Amino Acids
The mechanism by which amino acids are absorbed is conceptually identical to that of
monosaccharides. The lumenal plasma membrane of the absorptive cell bears at least four
sodium-dependent amino acid transporters - one each for acidic, basic, neutral and amino
acids. These transporters bind amino acids only after binding sodium. The fully loaded
transporter then undergoes a conformational change that dumps sodium and the amino acid
into the cytoplasm, followed by its reorientation back to the original form.
Thus, absorption of amino acids is also absolutely dependent on the electrochemical gradient
of sodium across the epithelium. Further, absorption of amino acids, like that of
monosaccharides, contributes to generating the osmotic gradient that drives water absorption.
The basolateral membrane of the enterocyte contains additional transporters which export
amino acids from the cell into blood. These are not dependent on sodium gradients.
Lecture 5
CLINICAL CORRELATION
Neutral Amino Aciduria (Hartnup Disease)
Thursday 6/10/2011
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2011
Thursday 6/10/2011
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2011
CLINICAL CORRELATION
A- -Lipoproteinemia
A-b-lipoproteinemia is an autosomal recessive disorder characterized by the absence of all
lipoproteins containing apo- -lipoprotein, that is, chylomicrons, very low density lipoproteins
(VLDLs), and low density lipoproteins (LDLs). Serum cholesterol is extremely low. This defect is
associated with severe malabsorption of triacylglycerol and lipid-soluble vitamins (especially
tocopherol and vitamin E) and accumulation of apo B in enterocytes and hepatocytes. The
defect does not appear to involve the gene for apo B, but rather one of several proteins involved
in processing of apo B in liver and intestinal mucosa, or in assembly and secretion of
triacylglycerol-rich lipoproteins, that is, chylomicrons and VLDLs from these tissues,
respectively.
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2011
Lecture 5
Thursday 6/10/2011
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2011
Lecture 5
Thursday 6/10/2011
Iron is absorbed by villus enterocytes in the proximal duodenum. Efficient absorption requires
an acidic environment.
Ferric iron (Fe+++) in the duodenal lumen is reduced to its ferrous form through the action of
a brush border ferrireductase. Iron is then co transported with a proton into the enterocyte via
the divalent metal transporter DMT-1. This transporter is not specific for iron, and also
transports many divalent metal ions.
Once inside the enterocyte, iron follows one of two major pathways:
Iron limiting states: iron is exported out of the enterocyte via a transporter
(ferroportin) located in the basolateral membrane. It then binds to the ironcarrier transferrin for transport throughout the body.
d) Copper
There appear to be two processes responsible for copper absorption:
i) a rapid, low capacity system and
ii) a slower, high capacity system, which may be similar to the two processes seen
with calcium absorption.
Many of the molecular details of copper absorption remain to be elucidated. Inactivating
mutations in the gene encoding an intracellular copper ATPase have been shown responsible
for the failure of intestinal copper absorption in Menkes disease.
A number of dietary factors have been shown to influence copper absorption. For example,
excessive dietary intake of either zinc or molybdenum can induce secondary copper
deficiency states.
e) Zinc
Zinc homeostasis is largely regulated by its uptake and loss through the small intestine.
Although a number of zinc transporters and binding proteins have been identified in villus
epithelial cells, a detailed picture of the molecules involved in zinc absorption is not yet in
hand.
A number of nutritional factors have been identified that modulate zinc absorption. Certain
animal proteins in the diet enhance zinc absorption. Phytates from dietary plant material
(including cereal grains, corn, rice) chelate zinc and inhibit its absorption. Subsistence on
phytate-rich diets is thought responsible for a considerable fraction of human zinc
deficiencies.
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Lecture 6
Sunday 9/10/2011
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2011
Lecture 6
Sunday 9/10/2011
This Na flux is electrogenic; that is, it is associated with an electrical current, and it
can be inhibited by the diuretic drug amiloride at micromolar concentrations
Microbial Fermentation
Fermentation is the enzymatic decomposition and utililization of foodstuffs,
particularly carbohydrates, by microbes
The large intestine does not produce its own digestive enzymes, but contains huge
numbers of bacteria which have the enzymes to digest and utilize many substrates.
In all animals, two processes are attributed to the microbial flora of the large
intestine:
1. Digestion of carbohydrates not digested in the small intestine
2. Synthesis of vitamin K and certain B vitamins
Cellulose is common constituent in the diet of many animals, including man, but no
mammalian cell is known to produce a cellulase. Several species of bacteria in the
large bowel synthesize cellulases and digest cellulose. Importantly, the major end
products of microbial digestion of cellulose and other carbohydrates are volatile fatty
acids, lactic acid, methane, hydrogen and carbon dioxide. Fermentation is thus the
major source of intestinal gas. Volatile fatty acids (acetic, proprionic and butyric
acids) generated from fermentation can be absorbed by diffusion in the colon.
Synthesis of vitamin K by colonic bacteria provides a valuable supplement to dietary
sources and makes clinical vitamin K deficiency rare. Similarly, formation of B
vitamins by the microbial flora in the large intestine is useful to many animals. They
are not absorbed in the large intestine, but are present in feces.
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Lecture 6
Sunday 9/10/2011
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2011
Lecture 6
Sunday 9/10/2011
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Lecture 6
Sunday 9/10/2011
Digestive Disorders
1. Stomach and Intestine
Peptic ulcers
Peptic ulcer is a general term that refers to ulcers occurring in the lower
esophagus, the stomach, or the duodenum (upper part of the small
intestine).
Sunday 9/10/2011
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2011
Caffeine
Caffeine seems to stimulate acid secretion in the stomach, which can
aggravate the pain of an existing ulcer. However, the stimulation of
stomach acid cannot be attributed solely to caffeine.
Alcohol
Although no proven link has been found between alcohol consumption
and peptic ulcers, ulcers are more common in people who have cirrhosis
of the liver, a disease often linked to heavy alcohol consumption.
Stress
Mucos HCO3 content creates a "micro-environment" around surface cells
to prevent acid damage, but its secretion is inhibited by adrenergic input
(prominent in stress!)
Acid and pepsin
It is believed that the stomach's inability to defend itself against the
powerful digestive fluids, hydrochloric acid and pepsin, contributes to
ulcer formation.
nonsteroidal anti-inflammatory drugs (NSAIDs)
These drugs (such as aspirin, ibuprofen, and naproxen sodium) make
the stomach vulnerable to the harmful effects of acid and pepsin.
2. Bile and the Biliary System
a. Gallstones (Cholelithiasis)
There are two major types of gallstones, which form due to distinctly different
pathogenetic mechanisms.
1. Cholesterol Stones
About 90% of gallstones are of this type. These stones can be almost
pure cholesterol or mixtures of cholesterol and substances such as mucin.
The key event leading to formation and progression of cholesterol stones is
precipitation of cholesterol in bile. There are clearly important genetic
determinants for cholesterol stone formation. There is also an important
gender bias in development of stones - the prevalence in adult females is two
to three times that seen in males and use of contraceptive steroids is a risk
factor for development of gallstones.
2. Pigment Stones
Roughly 10% of human gallstones are pigment stones composed of
large quantities of bile pigments, along with lesser amounts of cholesterol and
calcium salts. The most important risk factor for development of these
stones is chronic hemolysis from almost any cause - bilirubin is a major
constituent of these stones. Additionally, some forms of pigment stones are
associated with bacterial infections. Apparently, some bacteria release
deconjugate bilirubin, leading to precipitation as calcium salts.
Lecture 6
Sunday 9/10/2011
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b. Jaundice
Jaundice, is yellowing of the skin, sclera (the white of the eyes) and
mucous membranes caused by increased levels of bilirubin in the human
body. Usually the concentration of bilirubin in the blood must exceed 23mg/dL for the coloration to be easily visible. Jaundice comes from the
French word jaune, meaning yellow.
Causes of jaundice
When red blood cells
die, the heme in their
hemoglobin is converted
to bilirubin in the spleen.
The bilirubin is
processed by the liver, enters bile and is eventually excreted through
faeces.
Consequently, there are three different classes of causes for jaundice.
Pre-hepatic or hemolytic causes, where too many red blood cells are
broken down, hepatic causes where the processing of bilirubin in the
liver does not function correctly, and post-hepatic or extrahepatic
causes, where the removal of bile is disturbed.
1. Pre-hepatic
Pre-hepatic (or hemolytic) jaundice is caused by anything which causes
an increased rate of hemolysis (breakdown of red blood cells). Malaria
can cause jaundice. Certain genetic diseases, such as glucose 6phosphate dehydrogenase deficiency can lead to increase red cell lysis
and therefore hemolytic jaundice. Defects in bilirubin metabolism also
present as jaundice.
2. Hepatic
Hepatic causes include acute hepatitis, hepatotoxicity and alcoholic liver
disease. Jaundice commonly seen in the newborn baby is another
example of hepatic jaundice.
Lecture 6
Sunday 9/10/2011
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Neonatal jaundice
Neonatal jaundice is usually harmless: this condition is often seen in
infants around the second day after birth, lasting till day 8 in normal
births, or to around day 14 in premature births. Serum bilirubin normally
drops to a low level without any intervention required: the jaundice is
presumably a consequence of metabolic and physiological adjustments
after birth. Infants with neonatal jaundice are typically treated by
exposing them to high levels of colored light to break down the bilirubin.
Lecture 6
Sunday 9/10/2011
c. Cirrhosis
Cirrhosis is characterized anatomically by widespread nodules in the
liver combined with fibrosis. The fibrosis and nodule formation causes
distortion of the normal liver architecture which interferes with blood flow
through the liver. Cirrhosis can also lead to an inability of the liver to
perform its biochemical functions.
Causes of Cirrhosis
Alcoholic liver disease
Chronic viral hepatitis B, C and D
Chronic autoimmune hepatitis
Inherited metabolic diseases
Chronic bile duct diseases
Chronic congestive heart failure infections
Parasitic infections
liver inflammation that can be caused by fatty liver
long term exposure to toxins or drugs
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Lecture 6
Sunday 9/10/2011
3. Intestine
Diarrhea is an increase in the volume of stool or frequency of defecation.
It is one of the most common clinical signs of gastrointestinal disease,
but also can reflect primary disorders outside of the digestive system
There are numerous causes of diarrhea, but in almost all cases, this
disorder is a manifestation of one of the four basic mechanisms
described below.
1. Osmotic Diarrhea
Absorption of water in the intestines is dependent on adequate
absorption of solutes. If excessive amounts of solutes are retained in the
intestinal lumen, water will not be absorbed and diarrhea will result.
Osmotic diarrhea typically results from one of two situations:
Ingestion of a poorly absorbed substrate: The offending molecule
is usually a carbohydrate or divalent ion. Common examples include
mannitol or sorbitol, epson salt (MgSO 4) and some antacids (MgOH2).
Malabsorption: Inability to absorb certain carbohydrates is the most
common deficit in this category of diarrhea, but it can result virtually
any type of malabsorption. A common example is lactose intolerance
resulting from a deficiency in the brush border enzyme lactase. In
such cases, a moderate quantity of lactose is consumed (usually as
milk), but the intestinal epithelium is deficient in lactase, and lactose
cannot be effectively hydrolyzed into glucose and galactose for
absorption. The osmotically-active lactose is retained in the intestinal
lumen, where it "holds" water.
A distinguishing feature of osmotic diarrhea is that it stops after the
patient is fasted or stops consuming the poorly absorbed solute.
2. Secretory Diarrhea
Large volumes of water are normally secreted into the small intestinal
lumen, but a large majority of this water is efficiently absorbed before
reaching the large intestine. Diarrhea occurs when secretion of water into
the intestinal lumen exceeds absorption.
Many millions of people have died of the secretory diarrhea associated
with cholera. The responsible organism, Vibrio cholerae, produces
cholera toxin, which strongly activates adenylyl cyclase, causing a
prolonged increase in intracellular concentration of cyclic AMP within
crypt enterocytes. This change results in prolonged opening of the
chloride channels that are instrumental in secretion of water from the
crypts, allowing uncontrolled secretion of water.
Exposure to toxins from several other types of bacteria (e.g. E. coli heatlabile toxin) induce the same series of steps and massive secretory
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Lecture 6
Sunday 9/10/2011
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