03 September 2016

Lecture #7: THE ADRENAL GLAND



Structure


1.      Located at the top of the kidneys, fitting like caps.

2.      Made up of two portions:

a.      Adrenal cortex – composed of endocrine tissue
b.   Adrenal medulla – composed of neurosecreting tissue (catecholamines); inner portion.

THE ADRENAL CORTEX

Layers of secretory cells:


1.      Zona glomerulosa – outermost layer, directly under the outer connective tissue capsule of the adrenal gland; secretes mineralocorticoid.

2.      Zona fasciculate – middle layer; secretes glucocorticoid.

3.      Zona reticularis – inner layer; secretes small amounts of glucocorticoid and gonadocorticoids.

The corticosteroids

1.      Mineralocorticoids

a.      Has an important role in the regulatory process of mineral salts in the body.

b.      Aldosterone

(1)   Only physiologically important mineralocorticoid in human; primary function is maintenance of sodium homeostasis in the blood by increasing sodium reabsorption in the kidney.

(2)   Aldosterone also increases water retention and promotes the loss of potassium and hydrogen ions.

(3)   Aldosterone secretion is controlled by the renin–angiotensin mechanism and by blood potassium concentration.

Regulation of secretion of aldosterone

(1)   The initial step of aldosterone synthesis (cholesterol to pregnenolone) is under control of ACTH but the steps after pregnenolone synthesis are controlled by the renin–angiotensin system and the fluid volume.

(2)   If the sodium concentration and or plasma volume decreases, the proteolytic enzyme renin is released from the juxtaglomerular cells of the nephron into the plasma.

(3)   It has been hypothesized that baroceptors in the juxtaglomerular cells respond to changes in blood pressure. If the baroceptors are stretched because of an increase in blood pressure, renin secretion is inhibited, whereas a decrease in blood pressure stimulates renin secretions.

(4)   A circulating protein called angiotensinogen is cleaved by the renin to form angiotensin I.

(5)   Angiotensin I is modified by converting enzymes in the plasma and the lung tissue to become angiotensin II.

(6)   Angiotensinogen II stimulates the cells in the adrenal cortex to synthesize and release aldosterone.

(7)   In addition, angiotensin II causes constriction of the peripheral arteries, resulting in an increase in the blood pressure and an increased filtration rate in the kidney.

Assay methodology

(1)   RIA, available in kits (for renin and aldosterone)

(2)   For aldosterone determination, the position of the patient either lying down (supine) or upright must be noted for proper interpretation of results.

Normal values:         Upright = 5 – 30 ng/dl
                                    Supine = 3 – 10 ng/dl

(3)   The activity of renin is measured rather than the concentration of the enzyme itself and is based on the amount of angiotensin I produced from angiotensinogen.

Normal values:         Upright = 0.1 – 3.1 ng/hour/ml
                                    Supine = 1.6 – 7.4 ng/hour/ml

2.      Glucocorticoids

a.      Main glucocorticoid secreted by the zona fasciculata is cortisol, cortisone and corticosterone with cortisol the only one secreted in significant quantities.

b.      Affect every cell in the body.

c.       They are protein–mobilizing, gluconeogenic and hyperglycemic.

d.     Tend to cause a shift from carbohydrate catabolism to lipid catabolism as an enzyme source.

e.      Essential for maintaining normal blood pressure by aiding norepinephrine and epinephrine to have the full effect, causing vasocontriction.

f.        High blood concentration causes eosinopenia and marked atrophy in lymphatic tissues.

g.      Act with epinephrine to bring about normal recovery from injury produced by inflammatory agents.

h.      Secretion increases in response to stress.

i.        Except during stress response, secretion is mainly controlled by a negative feedback mechanism involving ACTH from the adenohypophysis.


Regulation of secretion of cortisol

a.      ACTH from the anterior pituitary is responsible for stimulating the release and synthesis of glucocorticoids from the zona fasciculata. The conversion of cholesterol to pregnenolone (the rate–limiting step) is under the control of ACTH as well as subsequent reaction that produce cortisol.

b.      Ninety percent of cortisol released into the blood circulates attached to CBG or albumin and has a half life of approximately 90 minutes.

c.       The secretion of cortisol is diurnal and is associated with a person’s sleep–wake cycle. A peak level of cortisol is usually observed between 6 and 8 am and a low level between 6pm and 12am. The concentration of cortisol in the plasma at 8pm is approximately 50% of the level at 8am.

d.     Changes in sleeping patterns or stress levels cause alterations in the secretion of ACTH and cortisol.

e.      Cortisol stimulates both gluconeogenesis and glycogenesis in the liver, resulting in increases in the plasma glucose.

(Click here for discussion about gluconeogenesis under Carbohydrate)
(Click here for discussion about Liver Function Test)

f.        The stimulation of gluconeogenesis in the liver occurs in the fasting state and in conjunction with cortisol’s inhibition of glucose uptake in peripheral tissues, causes the increase in plasma glucose.

g.      Since many amino acids are being used for gluconeogenesis in the liver, muscle uptake of amino acids is inhibited and protein catabolism increases, usually causing a loss of muscle tissue.

h.      Excess cortisol has been shown to decrease the amount of calcium that is absorbed from the intestine, initiating a decrease in plasma calcium. Plasma calcium concentrations are maintained by the body by removing calcium from storage in the bone tissue.

i.        Cortisol will stimulate lipolysis in adipose tissue so that the plasma concentration of glycerol and free fatty acids will increase.

j.        Cortisol in high concentrations will cause both anti–inflammatory and immunosuppressive actions. These actions of cortisol make it a valuable therapeutic agent in some types of diseases such as rheumatoid arthritis, systemic lupus erythematosus and multiple sclerosis.

GENERAL METHODS EMPLOYED IN STERIOD DETERMINATION

A.    Hydrolysis

1.      Acid hydrolysis (mineral acid method) – portion of a 24 hour urine sample is boiled under reflux in a specific length of time (10–60 minutes). Except for acid labile hormones, this method is preferred because of simplicity, speed and completeness of reaction.

2.      Enzymatic hydrolysis

a.      A portion of a 24 hour urine specimen is adjusted with buffer to the optimal pH of the enzyme employed.

b.      An adequate amount of the respective enzyme (beta–glucosidase to hydrolyze glucosidunates and sulfatase to hydrolyze sulfate conjugates) are added and the test sample incubated for 18–76 hours at 37oC).

c.       This method requires special attention regarding optimal concentration of enzyme, optimal pH of the enzyme, temperature and duration of incubation.

d.     The method is employed when the steroids are labile in strong acid solution (e.g. pregnanetriol, corticosteroid)

B.     Extraction

Organic solvents are used for extraction. Selection of solvents is based on polarity of the steroids.

Steroids with one or two oxygen (like androgens and estrogens) are low polarity and solvent used are chloroform, dichlormethane or ethyl acetate.

C.    Purification

This is necessary because a large number of urinary pigment chromogenic substances and other non–specific materials are invariable extracted along with the steroid and they will interfere in the final estimation of the steroid if they are not removed.

Strongly acid component migrate into aqueous layer and discarded. Neutral and phenolic steroids can be done in the same way. Phenolic steroids (estrogen) being acid in nature are readily extracted with an aqueous solution of hydroxide. After lowering the pH of the alkaline solution, estrogens are re – extracted with a suitable solvent like diethyl ether and are processed further for final estimation.

D.    Estimation

1.      Colorimetric method
2.      Flurometric method
3.      Gas chromatography

Urinary metabolites of cortisol quantitated

1.      17–hydroxycorticosteroids – have hydroxyl groups at C–21 and C–17 and a keto group at C–20; this configuration is known as dihydroxyacetone side chain. These compounds are measured using the Porter–Silber reaction.

2.      17–ketogenic steroids group as:

Group I          :           cortisol, cortisone, 11–deoxycortisol, tetrahydro S
Group II         :           cortols and cortolones
Group III        :           pregnanetriol and 11–oxygenated derivatives
Group IV       :           17–hydroxyprogesterone, 17–hydroxypregnenolone

·         These compounds are measured using the Zimmerman reaction

Interference in 17–ketosteroid determination

a.      Cortisol
b.      Dexamethasone
c.       Equanil
d.     Librium
e.      Quinine
f.        Seconal
g.      Thorazine
h.      Serpasil
i.        Nodular
j.        Paraldehyde


The Porter–Silber reaction

17–hydroxycorticosteroids and phenylhydrazine are incubated at 60oC for 30 minutes. A characteristic yellow pigment is formed that can be measured in a spectrophotometer at 410 nm.


The Zimmerman reaction (Appleby method)

1.      Treatment of sodium bismuthate to break the 17–hydroxyl bond and replace it with a ketogroup at position 17.

2.      The 17–ketosteroid is then quantitated by producing a color reaction with dinitrobenzene in the presence of alcohol and potassium hydroxide.

3.      A reddish purple pigment is formed and readings are taken in a spectrophotometer at 480, 520 and 580 nm.


Biosynthesis of Adrenal Hormone

The biosynthesis of the adrenal steroids is a multienzyme (cytochrome P–450 oxygenase) process that takes place in the cytoplasm and mitochondria of the cortex endocrine cells. Synthesis of the various steroid hormones start at the mitochondria with the conversion of cholesterol, primarily from the plasma lipoprotein, to pregnenolone. The pregnenolone must be transported out of the mitochondria and to the various intracellular sites for the process of hormone synthesis to continue.

In addition, this is the rate–limiting step in the synthesis of adrenal steroids and is primarily under the control of adrenocorticotropic hormone (ACTH). At this point, the pathways diverge to produce the different hormone. The individual layers of the adrenal cortex contain the enzyme necessary for each of the respective hormones produced there. For example, the zona glomerulosa has the enzyme 18– hydroxysteroid dehydrogenase that is necessary for the production of aldosterone but lacks the enzyme 11–beta–hydroxylase necessary for cortisol synthesis. This layer also lacks the enzyme 17–alpha–hydroxylase that is required for the production of androgens.


Metabolism of adrenal hormone

The major site of metabolism for the steroid hormone is the liver. The rate of metabolism by the liver is dependent on whether the hormone is bound to a protein carrier. A higher percentage of aldosterone is not protein bound is removed from circulation and metabolized by the liver more rapidly than cortisol, which is mostly bound to a protein carrier (transcortin, also called corticosteroid–binding protein, CBG).

Most steroids excreted in urine must be conjugated by the liver to either glucuronic acid or sulfates to form soluble compounds. The metabolism of cortisol begins in the liver with a delta reductase enzyme and dehydrocortisol is produced. Dehydrocortisol is not excreted in urine and is further metabolized by 3–alpha– hydroxysteroid dehydrogenase. The metabolites formed from this reaction are the major (50%) excretory compounds of cortisol (tetrahydrocortisol, tetrahydrocortisone). Approximately 30% of urinary metabolites of cortisol are cortrol and cortolone and are formed by the hydrogenation of the C–20 ketone group. In normal patients, very little urinary free cortisol is found. Conversely, relatively large amounts of urinary free cortisol are seen in patients with Cushing’s syndrome.


Clinical significance of adrenal cortex function

1.      Primary Mineralocorticoid Excess

a.      Primary Aldosteronism

2.      Syndrome with Deoxycorticosterone (DOC) Excess

a.      17–alpha–hydroxylase deficiency
b.      11–beta–hydroxylase deficiency
c.       Androgen–estrogen–producing adrenal tumors
d.     Syndrome of Primary Cortisol Resistance

3.      Syndrome with Secondary Mineralocorticoid Excess

a.      Hypertension

4.      Syndrome of Glucocorticoid Excess and Hypertension

a.      Cushing’s syndrome

b.      Other hypertensive syndromes with pseudorhyperaldosteronism

(1)   Secondary Mineralocorticoid Deficiency
(2)   11–beta–hydroxysteroid dehydrogenase deficiency
(3)   Chronic ingestion of licorice
(4)   Liddle’s syndrome
(5)   Arnold–Healy–Gordon syndrome


THE ADRENAL MEDULLA

Structure

1.      Located at the middle portion of the adrenal gland

2.      Chromaffin cells or pheochromocyte

a.      Large, ovoid, columnar cells arranged in clumps or cords around blood vessels.

b.      Their granules turn brown when stained with chromic acid due to the oxidation of epinephrine and norepinephrine to melanin.

c.       These cells have large nuclei and well–developed Golgi complex.

d.     They contain large numbers of vesicles or granules containing catecholamine. Vesicles containing norepinephrine are darker than those containing epinephrine.

e.      The reason for the high production of epinephrine is the presence of the enzyme phenylethanolamine–N–methyltransferase (PNMT).
  


Schema of embryonic development of adrenergic cells


Hormones of the adrenal medulla

Catecholamine is a collective term used to describe the hormones of the adrenal medulla. The adrenal medulla is a highly innervated tissue with many neurons that appear to be in direct contact with chromaffin cells. Catecholamine hormone release can be stimulated by the action of these neurons.

1.      Dopamine

a.      Appear in highest concentration in the central nervous system, where if functions as a neurotransmitter.

b.      In hypothalamus, it helps to control the synthesis of prolactin from the pituitary. When dopamine is present, prolactin is inhibited and when dopamine is absent, prolactin is freely secreted.

c.       The final metabolite of dopamine is homovanillic acid (HVA).


2.      Norepinephrine

a.      The highest concentration of norepinephrine is found in the brain (CNS) and to a lesser degree in the sympathetic nervous system (SNS) where it functions as neurotransmitter.

b.      Acting primarily through the alpha adrenergic receptors, norepinephrine can cause many varied responses. These responses include such physiologic actions as vasoconstriction of the small vessels in the skin or relaxation of the smooth muscle of the gastrointestinal tract.

3.      Epinephrine

a.      The major catecholamine produced by the chromaffin cells of the adrenal medulla.

b.      It is often called the “flight or fight” hormone since it is released in response to physiologic (injuries) or psychological (stress, anxiety, threats).

c.       Epinephrine secretion causes increased heart rate, blood pressure, respiration and metabolic rate.

d.     In addition, it stimulates glucogenolysis in the liver and skeletal muscle that leads to increase in the plasma glucose level.


Synthesis of catecholamine

The synthesis of catecholamine can be measured by the following procedures

1.      Conversion of tyrosine to dopa

The catecholamines are synthesized from tyrosine, which may be derived from ingested food or synthesized from phenylalanine in the liver. Tyrosine circulates at a concentration of 1–1.5 mg/dl of blood. It enters neurons and chromaffin cells by an active transport mechanism and is converted to dihydroxyphenylalanine (dopa). The reaction is catalyzed by tyrosine hydroxylase, which is transported via axonomal flow to the nerve terminal.


2.      Conversion of dopa to dopamine

This is through the action of the enzyme dopa decarboxylase. This enzyme is found in all tissues, with the highest concentrations in the liver, kidney, brain and vas deferens. Competitive inhibitors of dopa decarboxylase such as methyldopa are converted to substances that are then stored in granules in the nerve cell and released in place of norepinephrine.


3.      Conversion of dopamine to norepinephrine

This is through the action of the enzyme dopamine β–hydroxylase. This enzyme when newly synthesized is incorporated directly into storage vesicles that take up, synthesize and store catecholamines. The membranes of these vesicles contain dopamine β–hydroxylase, ATPase, cytochrome P–561: NADH reductase. Dopamine β–hydroxylase is also found within the granules and is released with norepinephrine during secretion. Inhibitors of the enzyme, such as disulfuramic and picolinic acid, have no clinical importance.


4.      Conversion of norepinephrine to epinephrine

PNMT catalyzes the N–methylation of norepinephrine to epinephrine, using 8–adenosylmethionine as a methyl donor. It is found only in the adrenal medulla and in a few neurons in the central nervous system. The enzyme is found in the cytosol. Norepinephrine leaves the granule and after methylation reenters different granules. This enzyme is induced by the high levels of glucocorticoids found in the adrenal medulla. The conversion of dopamine to epinine is catalyzed by a non–specific N–methyltransferase


Metabolism of catecholamines
                       


Catecholamines that are secreted into the plasma have a short half–life of about 1 to 2 minutes. They are rapidly remove from the circulation by the liver and kidneys or taken up by sympathetic neurons.

Two enzyme systems are responsible for the breakdown or inactivation of the catecholamine hormones. Monoamine oxidase (MAO) deaminates the catecholamines and is present in many tissues throughout the body.

The second enzyme, catechol–o–methyl–transferase (COMT), methylates a hydroxyl group on the aromatic ring structure of the catecholamine molecule. COMT is found in highest concentration in kidneys and liver. The methylation reaction of this enzyme produces metaneprine from epinephrine and normetanephrine from norepinephrine.

Both metanephrine and normetanephrine can be conjugated with sulfate or glucuronide and excreted in the urine. The final metabolite of both epinephrine and norepinephrine is vanillylmandelic acid (VMA).
  

Receptors

The action of the catecholamine hormones is mediated through adrenergic receptors. There are two alpha receptors and two beta receptors. These receptors are found throughout the body in most tissues. Some tissues have both alpha and beta receptors present, whereas other tissues may have only one type.

Both epinephrine and norepinephrine interact with these receptors, although the receptor affinity for the two hormones may be different. The alpha–1 receptor uses calcium and phosphatidyl inositol for its second messengers. The alpha–2, beta–1 and beta 2–receptors are cAMP as their second messenger.


Regulation of secretion of catecholamines


Adrenal medullary catecholamine secretion is increased by exercise, angina pectoris, myocardial infarction, hemorrhage, ether anesthesia, surgery, hypoglycemia, anoxia and asphyxia and many other stressful stimuli.

The rate of secretion of epinephrine increases more than that of norepinephrine in the presence of hypoglycemia and most other stimuli. However, anoxia and asphyxia produce a greater increase in adrenal medullary release of norepinephrine than is observed with other stimuli.

Secretion of the adrenal medullary hormones is mediated by the release of acetylcholine from terminals of preganglionic fibers. The resulting depolarization of the axonal membrane triggers an influx of calcium ions. The contents of the storage vesicles, including the chromograms (storehouse of catecholamines) and soluble dopamine beta–hydroxylase, are released by exocytosis by the calcium ion increase. Membrane bound dopamine beta–hydroxylase is not released.

Tyramine, however, releases norepinephrine primarily from the free store in the cytosol. Cocaine and monoamine oxidase inhibitors inhibit the effect of tyramine but do not affect the release of catecholamines by nervous stimulation. The rate of release in response to nerve stimulation is increased or decreased by a wide variety of neurotransmitter acting at specific receptors on the presynaptic neuron. Norepinephrine has an important role in modulating its own release by activating the alpha receptors on the presynaptic membrane.

Alpha–2 receptor antagonists inhibit this reaction. Conversely, presynaptic beta receptors enhance norepinephrine release, whereas beta receptor blockers increase it. The accumulation of excess catecholamine that is not in the storage granules is prevented by the presence of intraneuronal monoamine oxidase.


Laboratory evaluation of adrenomedullary function

1.      Dopamine

a.      Assay methodology: radioenzymatic kit method using plasma

Utilizes the enzyme COMT to transfer a H–methyl group to the catecholamine. All catecholamine present in the sample are H– methylated. The catecholamines is extracted from the sample and are separated by thin layer chromatography. The amount of radioactivity in each of the chromatography spots can be measured and the individual catecholamine quantitated.

Normal value:    0 – 83 pg/dl

Precaution in specimen collection

(1)   Specimen should be collected by catheterization since venipuncture tend to increase catecholamine

(2)   The blood drawing tubes should contain sodium thiosulfate and EDTA.

(3)   Blood samples must be placed on ice immediately after drawing

(4)   The blood should be separated with a refrigerated centrifuge as soon as possible after drawing and the plasma frozen at –70oC until the analysis is performed.


b.     Urine HVA using GC or HPLC

Precaution in specimen collection

(1)   A 24 hours specimen is collected and 10 ml of concentrated HCl is added as preservative.

(2)   During the collection time, the urine container should be kept in refrigerator.

(3)   Once the 24 hour urine specimen is returned to the laboratory, the toal volume should be recorded and the specimen is aliquoted.

(4)   The pH of all aliquots should be adjusted to about pH 3 to 4 using 6M HCl.

Normal values:               <15 mg/24 hours


2.      Norepinephrine and epinephrine

a.      Using plasma sample

(1)   The radioenzymatic method described previously for dopamine using COMT and H–methylation of the catecholamine followed by TLC can also be used here.

Normal values:    Epinephrine = 0 – 62 pg/ml
                                Norepinephrine = 111 – 603 pg/ml

(2)   Ethylenediamine (EDA) Fluorometric method

(a)   Extracting the catecholamine from the plasma first with an alumina column followed by an Amberlite CG–50 column.

(b)   EDA is added to the collected eluate, where it combines with the catecholamine

(c)    Three fluorescent products are obtained from norepinephrine and one from the epinephrine.

(d)  These products are measured in a fluorometer at two different wavelengths (emission 510 and 580 nm, activation 420) and concentrations of the hormones are calculated.

Normal values:               Epinephrine = 140 – 300 pg/ml
                                          Norepinephrine = 360 – 800 pg/ml

Assay interference:        Penicillin, coffee, tea

b.     Using urine sample

(1)   Urine vanillylmandelic acid and colorimetric method

(a)   Hydrolysis with HCl to destroy conjugates is the initial step in the process.

(b)   Solvent extraction with ethyl acetate separates the VMA into the organic layer.

(c)    Further purification of the mixture is accomplished by reextracting the VMA into an aqueous potassium carbonate solution.

(d)  Treatment with sodium metaperiodate oxidizes VMA to vanillin which can be quantitated spectrophotometrically.

Normal values:         2 – 7 mg/24 hours


Interference in the determination

(a)   Falsely decreased values

pH above 3 decomposes VMA, salicylates, clofibrate and L–dopa, clonidine, disulfram, hydrazine derivatives, imipramine, MAO inhibitors, morphine, phenothiazines and some radiography agents.


(b)  Falsely elevated values

Bananas, coffee, tea, vanilla, chocolates, salicylates, disulfram, nalidixic acid, oxytetracycline, pthalein dyes, insulin, isoproterenol, L–dopa, lithium, reserpine


(2)   Fluorometric assay

(a)   Adjust the urine pH to alkaline range. Both epinephrine and norepinephrine exist as neutral compounds at pH values above 8 since the amine groups on these molecules are not protonated. These forms of molecules then bind to alumina or resin ion– exchange columns.

(b)   After washing to remove other materials, the catecholamine is eluted with dilute acid.

(c)    Oxidation with ferricyanide converts the catecholamine to cyclic derivatives. Addition of trihydroxyindole derivatives to fluorescent forms.

Normal values:               Epinephrine = 0 – 15 ug/24 hours
                                          Norepinephrine = 0 – 100 ug/24 hours
                                          Dopamine = 65 – 400 ug/24 hours

Interference in the assay:

Falsely elevated values is seen in pH values below about 8.5 will lead to lowered extraction efficiency, prolonged refrigeration sometimes discolors the sample causing increased blank values.

(3)   Urine metanephrine using Pisano method

(a)   Hydrolysis with HCl breaks down all conjugates, allowing more uniform extraction of all fractions.

(b)   The solution containing the metanephrine mixture is poured onto an ion–exchange column which loosely binds the metanephrine but allows other components to pass through and be removed from the reaction mixture.

(c)    Elution with ammonium hydroxide washes the metanephrine off the column.

(d)  Conversion to vanillin is accomplished through periodate oxidation.

Normal values:               Metanephrine = 74 – 297 ug/24 hours
                                          Normetanephrine = 105 – 354 ug/24 hours
                                          HVA = 66 – 222 ug/24 hours

Interference in the assay:

Falsely elevated values: chlorpromazine, imipramine, phenothiazines, methyldopa, tetracycline


Clinical significance

1.      Increased catecholamine level is seen in

a.      Pheochromocytoma
b.      Neuroblastoma
c.       Essential hypertension
d.     Hypothyroidism
e.      Diabetic acidosis
f.        Cardiac disease
g.      Burns
h.      Septicemia
i.        Depression

2.      Decreased catecholamine level is seen in

a.      Hyperthyroidism
b.      Long–term diabetes


******  PHEOCHROMOCYTOMA ******

Pheochromocytoma is a rare tumor of the chromaffin cells. Adrenal chromaffin cell tumors make up about 90% of all pheochromocytoma. The adrenal tumors may secrete large amounts of epinephrine, norepinephrine or varying combinations of both. Pheochromocytomas found in areas other than the adrenal medulla secrete mostly norepinephrine. Most pheochromocytomas occur in adults and in each sex about equally.

Signs and symptoms

1.      Symptoms during or following paroxysms

a.      Headache
b.      Sweating
c.       Forceful heartbeat with or without tachycardia
d.     Anxiety or fear of impending death
e.      Tremor
f.        Fatigue or exhaustion
g.      Nausea and vomiting
h.      Abdominal or chest pain
i.        Visual disturbances

2.      Symptoms between paroxysm

a.      Increased sweating
b.      Cold hands
c.       Weight loss
d.     Constipation

The hypertension seen in connection with a pheochromocytoma in an alpha receptor effect related to the excess of norepinephrine produced. The hypertension can be sustained or paroxysmal. The hypermetabolic features seen in pheochromocytoma resemble those seen in patients with hyperthyroidism. These include heat intolerance, tremor, weight loss, palpitations and increase BMR.

In contrast to hyperthyroidism, in which the patient has hyperthyroidism, patients with pheochromocytoma have constipation. The increased levels of catecholamine reduced gastrointestinal motility and increase sphincter tone, which also lead to nausea, abdominal pain and possibly obstruction.

Pheochromocytoma may be misdiagnosed since the symptoms are similar to those of several other disorders. When diagnosed correctly, they are easily treated surgically. If left untreated, the patient usually dies.

Catecholamine has a tendency to block insulin release, increase gluconeogenesis and cause fatty acid mobilization. These actions can elevate serum glucose levels. Some patients have hypokalemia owing to their diuretic therapy or an elevation of plasma renin and aldosterone.



******  NEUROBLASTOMA  ******


Neuroblastoma is a rare form of malignant tumor of cells that are neuron precursors found in infants and children. The primary tumor often is in or near the adrenal gland and is not found until after the tumor has metastasized to another site such as the liver. Neuroblastoma secretes sporadic increases in catecholamine hormones. The symptoms due to the catecholamines may be transient hypertension, sweating, tachycardia and headaches. Generally, the most persistent symptom is due to tissue damage from the secondary metastatic sites, that is, the liver or lymph nodes.

Problems encountered in collecting sample

1.      Collection of a 24 hours specimen from infants and small children.

2.      Seventy to eighty percent of children show increases in urinary VMA, but the increases vary from just slightly above normal to about ten times the normal level.



  



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