EN1 and 2
EN1 and 2
EN1 and 2
● related tests: T3 and T4 Hashimoto Thyroiditis ↓/N ↓ /N ↑ N/↓ ↓ N/↓ N
○ Adults: 0.5-4.7 μunits/L Nonthyroidal illness ↓ N/↓ V V N/↑ N N
○ Pregnancy (1st): 0.3-4.5 μunits/L hyroid Hormone
T
○ Pregnancy (2nd): 0.5-4.6 μL Resistance ↑ ↑ N/↑ ↑ ↑ ↑ N
○ Pregnancy (3rd): 0.8-5.2 μL
TSH Immunoassay PARATHYROID GLAND
● S econd-Generation TSH Immunometric Assays ● Located near the thyroid capsule,
○ with detection limits of 0.1 mU/L ○ sometime within the thyroid gland
○ screen for hyperthyroidism ○ Most of the people they have four parathyroid gland but in some they have
● Third-generation TSH chemiluminometric assays eight or as few as two
○ with detection limits of 0.01 mU/L, (euthyroidism and hyperthyroidism) ○ Secrete PTH
○ routinely used to monitor and adjust thyroid hormone replacement therapy ● Samples endocrine gland in the body
○ screen both hyperthyroidism and hypothyroidism. ● four tiny glands attached to the thyroid
○ The sensitivity of this has lead to the ability to detect what is turned ● releases PTH
subclinical disease or the mild degree of thyroid dysfunction ○ The primary role of the PTH is to prevent hypocalcemia
Increased TSH Decreased TSH ○ It regulates the blood calcium
Primary Hypothyroidism Primary Hyperthyroidism ○ Preserve calcium and phosphate within the normal range
Hashimoto’s Thyroiditis Secondary and Tertiary Hypothyroidism ○ Promotes bone resorption → release calcium into the bloodstream
Thyrotoxicosis due to Pituitary Tumor Treated Grave’s Disease ○ PTH also increased renal reabsorption of calcium
TSH Antibodies Euthyroid Sick Disease ○ Stimulate the conversion of inactive vitamin D to vitamin D3
Thyroid Hormone Resistance Over Replacement Hormone in ○ actions directed to bone, kidney and intestines
Hypothyroidism ○ controls calcium and phosphate metabolism with the help of calcitonin
Types of cells:
Anti-TSH Receptor Autoantibody
● Chief cells
for diagnosis ofGrave’s disease
● ○ synthesize and secrete hormone PTH
● detects autoantibodies that interfere with the binding of TSH to TSH receptor ● Oxyphil cells
s
● erum + TSH receptor + I125 labelled TSHtracer ○ non secretory cell
● amount of free tracer is measured ○ seen only after puberty
○ lower than 9 U/L DISEASES ASSOCIATED WITH HORMONES OF THE PARATHYROID GLAND
Thyrotropin Releasing Hormone (TRH) Stimulation Test
It measure the relationship between TRH and TSH secretion
● Hyperparathyroidism
● Used to differentiate euthyroid and hyperthyroid patient who both had
undetectable TSH level Primary Hyperparathyroidism
● Also be helpful in the detection of the thyroid hormones resistance ● Physiologic defect lies with theParathyroid gland
syndrome ● Most common causeof hypercalcemia
● injection of TRH and measurement of the output of TSH ● Is due to the presence of a functioningparathyroidadenoma
○ Dose needed TRH: 500 ug by IV ● Is accompanied withphosphaturia
○ Increase: primary hypothyroidism ● If it goes undetected,severe demineralizationmayoccur (osteitis fibrosa
○ Decrease: hyperthyroidism cystica)
● used in the diagnosis of combined pituitary- thyroid disorders ● Lab Results:
● differentiates 2° hypothyroidism and 3°hypothyroidism ○ PTH increased
Other Laboratory Tests: ○ Ionized Ca increased
● Radioactive iodine (RAI) uptake: ○ Hypercalciuria
○ based on the ability of the thyroid to concentrate, convert and release4 I2 ○ Hypophosphatemia (fasting state)
○ Used to measure the ability of the thyroid gland to trap iodine Secondary Hyperparathyroidism
○ Helpful in establishing the cause of hyperthyroidism ● Develops in response to decrease serum calcium
○ High uptake = indicates that there is metabolically active hormone + ● There isdiffuse hyperplasiaof all 4 glands
Low or TSH deficiency = autonomous thyroid activity ● The patient develops severebone disease
● Thyroid Blocking Immunoglobulin (TBI): ● Causes:vit. D deficiency and chronic renal failure
○ based on the thyroid hormone transport system indirectly measuring the ● Lab results:
amount of TBG ○ PTH increased
● Protein Bound Iodine Test (PBI): ○ Ionized calcium decreased
○ based on thyroid hormone concentration representing the organic fraction Tertiary Hyperparathyroidism
of blood iodine that precipitates with serum proteins
● Itoccurswithsecondary hyperparathyroidism
○ A laboratory test that indirectly assesses the thyroid function by
● Develops autonomous function of the hyperplastic parathyroid glandsor of
measuring the concentration of iodine bound to proteins circulating in the
parathyroid adenoma
bloodstream
● The phosphate levels are normal to high;Calcium phosphatesprecipitates in
● Basal Metabolic Rate (BMR):
soft tissues
○ based on metabolic response measuring the O2 consumption in the
Hypoparathyroidism
resting fasting state
● Is due toaccidental injury to the parathyroid glands(neck) during surgery-
Summary: Thyroid Function Tests
post-surgical cause
1. TRH Stimulation Test
● Other cause:autoimmuneparathyroid destruction
2. TSH Test
● Individual are unable to maintain calcium concentration in the blood without
3. Radioactive Iodine Uptake (RAIU)
calcium supplementation
4. Thyglobulin (Tg) assay
● In hypoparathyroidism, the distal convoluted tubules reabsorbsbicarbonate as
5. Reverse T3 (rT3)
well as phosphateresulting in acidosis
6. Free Thyroxine Index (FT4I or T7)
● PTH normally interferes with bicarbonate reabsorption in the PCT; therefore, the
7. Total T3 (TT3), FT3, FT4
renal tubular bicarbonate threshold tends to be in increased in
8. T3 Uptake Test
hypoparathyroidism
L
● ow PTH causes elevated bicarbonate reabsorption-alkalosis Glucagon Immunoassay (RIA)
● The best method for PTH measurement involves the use ofantibodies that 125
● glucagon competes with I tracer for binding sites
detect both the amino terminal fragment and intact PTH
125
Laboratory method for PTH: ● a mount of I is measured and is inversely proportionalto the concentration of
● PTH level measurement: glucagon
○ overnight fasting ● Fasting: 60-200 pg/mL
■ Intact PTH: 10-65 pg/mL
■ PTH N-terminal (includes intact PTH): 8-24 pg/mL
■ PTH C-terminal (includes C-terminal, intact PTH, and midmolecule):
50-330 pg/mL
● related tests: Calcium, Phosphorus and Creatinine
Pancreas
● Organ located in the abdomen
● Essential role: converting food into fuel for the body's cell
● Two main function
○ Exocrine organ – that help indigestion
○ Endocrine – regulate blood sugar
● lying immediately beneath the stomach
● both an exocrine and an endocrine gland
Types of tissues:
● Acini
○ secretes digestive juices into the intestine
● Islets of Langerhans
○ secretes hormones directly into the blood
Hormones in Pancreas
● Glucagon: glycogenolysis and gluconeogenesis
● Insulin: glycogenesis, glycolysis, lipogenesis
● Somatostatin
DISEASES ASSOCIATED WITH HORMONES OF THE PANCREAS
Diabetes mellitus
d
● e ficiency of insulin or defects in insulin receptors
● Increase in blood glucose level
● Causes: due to stress, severe infection, dehydration or pregnancy,
pancreatectomy, haemochromatosis
● FBS – high >126 mL/d
Hyperinsulinism
● It refers to above normal level of insulin in the blood of a patient
○ The secretion of insulin are closely related to the level of glucose in the blood
○ The insulin of an individual is maybe normal but it can be low or high to
another individual
❖ The secretion is depend on level of the blood glucose of the individual in
the body
❖ For a certain individual we should check what is the normal level of
his/her insulin before it considered that there is hyperinsulinism
● hypersecretion of insulin
○ Insulin is the primary hormone responsible for the entry of glucose in the cell
❖ Hypoglycemic agent → release by beta cell
● may be due to a tumor, insulinoma
Glucagonoma
● hypersecretion of glucagon by a tumor
○ Glucagon is a primary hormone responsible for an increase in glucose level
○ It is a hyperglycemic agent → alpha cell → release during stress and fasting
state
○ Work with insulin to control the amount of sugar in the blood
○ If there is an tumor they produced large amount of glucagon
○ If high levels continuously it can create severe and life-threatening symptoms
Somatostatinoma
● hypersecretion of somatostatin by a tumor
● Somatostatin which is produced by delta cells
○ Inhibits the action of insulin and glucagon
Laboratory Measurement of some hormones secreted by the Pancreas
ake note: after the cholesterol was converted into pregnenolone you will now have
T
here the different hormones
Cortisol
● Principalglucocorticoid
● Synthesis is regulated byACTH
● Critical to hemodynamic and glucose-homeostasis
○ The glucocorticoids they maintain blood glucose by inducing lipolysis and
gluconeogenesis
○ They have anti-inflammatory and immunosuppressive action
Hormones ● Mostly bound to glycoprotein(transcortin)
Adrenal Cortex Adrenal Medulla ● Stimulateslipolysis and gluconeogenesisin the liver
● The only adrenal hormone thatinhibitsthe secretion of ACTH**
Cortisol Norepinephrine
● Therapeutic agent forRA, SLE, MS**
Aldosterone Epinephrine
● Secretion is diurnaland is associated with sleep-wakecycle (highest at 8-9am
Weak androgens Dopamine
and lowest 10pm-12am)
Adrenal Cortex
● Major site of steroids hormone production
○ Cholesterol is the parent cell of all steroids hormones
● The outer region of the adrenal gland secreting thesteroid hormone
● The secretion of adrenal glucocorticoids and androgens is regulated by
Adrenocorticotropic hormone (ACTH), which is under the control of the
hypothalamiccorticotropin-releasing hormone(CRH)
● Mineralocorticoidssecretion is controlled by theRAAS
● Cortical hormones are composed of CPPP(cyclopentanoperhydrophenanthrene)
3 Layers of Adrenal Cortex
1. Zona Glomerulosa(outermost layer)- 10%
○ Principal source of mineralocorticoids he cortisol can be found or released in the F-zone, In the F-zone the main steroid is
T
■ Salt regulation the cortisol and the regulator is the ACTH. the function is for blood pressure and
2. Zona Fasciculata(middlemost layer) -75% glucose homeostasis
○ Site of glucocorticoid synthesis Syndrome:
○ Also synthesize unsulfated DHEA Low cortisol → adrenal insufficiency → symptoms: hypotension, hypoglycemia,
3. Zona Reticularis(innermost layer) -15% weight loss, weakness
○ Produces androstenedione and dehydroepiandrosterone (weak androgens) Increase cortisol → hypercortisolism → symptoms: hypertension, hyperglycemia,
3 Layers of Adrenal Cortex central obesity, weakness
● Aldosterone
○ low cytoplasm to nuclear ratio and small nuclei withdense chromatinwith
intermediate lipid inclusions.
● Cortisol-
○ high cytoplasm to nuclear ratio, lipid laden with“foamy”cytoplasm.
● DHEAS
○ lipid deficient buthas lipofuscindeposit sharply demarcated cells
For the measurement of aldosterone Adrenal Insufficiency
● Serum, plasma, saliva and urine may be used, ● L ow baseline cortisol levels (8:00 am, supine) and an elevated ACTH greater
○ blood samples should be drawn at8 am than 200 pg/mL(PRIMARY)
● Urine free cortisollevels are sensitive indicators of adrenal hyperfunction ● Lower serum concentrations of ACTH and cortisol(SECONDARY)
(endogenous corticolism-24 hour urine collection) ○ Usually suggested in patient who have near normal response in cosyntropin
Urinary Metabolites test 200 pg/mL but abnormal response to metyrapone test
● The liver degrades all glucocorticoids to metabolites excreted in urine Cosyntropin Test
1. 17-hydroxycorticosteroid ● determines the capacity of the adrenal gland to increase hormone production in
● Measured byPorter-Silber Method response to stimulation.
● Reagents: Phenylhydrazine in H2SO4 + acohol Metyrapone Test
2. 17-ketogenic steroids ● block 11β-hydroxylase, increasing 11-deoxycortisol (>7 μg/dL) while cortisol
● Measured byZimmermann Reaction(reddish purple) decreases (<5 μg/dL).
● Reagent: Meta-dinitrobenzene In both test there will be synthetic stimulator of cortisol and aldosterone secretion that
● Oxidation Procedure: Norymberski (Na bismuthate) will be given
Clinical Disorders Primary Hypocortisolism
● Hypercortisolism ● Due to decreased cortisol production- 90% destruction of the adrenal cortex;
● Hypocortisolism aldosterone deficiency; excess ACTH release
○ Primary Hypocortisolism Disorder:
○ Secondary Hypocortisolism Addison’s Disease
● Congenital Adrenal Hyperplasia ○ Hypotension, hyponatremia, hyperkalemia, weight loss, hyperpigmentation)
○ 21-hydroxylase deficiency ○ Due to autoimmune adrenalitis, tuberculosis, hemorrhage, HIV/AIDS infection
○ 11ß-hydroxylase deficiency Screening Test:
○ 3ß-hydroxysteroid dehydrogenase isomerase deficiency ○ ACTH Stimulation Test
○ C-17,20-lyase/ 17α-hydroxilase deficiency ■ Increased ACTH and decreased cortisol and aldosterone
Hypercortisolism Secondary Hypocortisolism
● Also known as thesecondary adrenal insufficiency
Cushing’s Syndrome ● Due to hypothalamic-pituitary insufficiency with loss ofACTH
● Primarily caused by excessive production ofcortisol and ACTH ● No problem with mineralocorticoid secretion;absence of hyperpigmentation
● Caused by overuse ofcorticosteroid ● Lab Test:ACTH Stimulation Test
Signs and Symptoms: ○ (+) result- delayed in response to stimulation test.
● Weight gain but with thin extremities“buffalo hump”, hyperglycemia, thinning of ○ Decreased ACTH and cortisol
the skin, poor wound healing, hypertension, hypercholesterolemia, decreased Procedure of ACTH Stimulation Test
WBC ● Also known asCosyntropin Stimulation Test
Screening Test ● Also differentiates secondary adrenal insufficiency (decreased or no ACTH
● 24 hour urinary free cortisol (inc >120 ug/day) response) from tertiary adrenal insufficiency (increased ACTH response)
● Overnight dexamethasone suppression test-most widely used salivary ○ stimulation test that can be used to distinguished hypothalamic or tertiary
cortisol adrenal insufficiency and pituitary or secondary adrenal insufficiency
Confirmatory Test ● Cosyntropin is a synthetic cortisol and aldosterone stimulator
● Low-dose dexamethasone suppression test- ● It requires administration of250 ugof Cosyntropin IV or IM
● Midnight plasma cortisol (> 5.0 ug/dL) Metyrapone Test
● CRH stimulation test ● Measures the ability of the pituitary gland to respond to declining levels of
circulating cortisol, thereby secretes ACTH
● Is used as an alternative diagnostic or confirmatory test fortertiary adrenal
insufficiency
● 500 to 750 mg of metyrapone is orally administered every 4 hours for 24 hours
● Performed only if the ACTH stimulation test gives normal result
● (+) result: decreased ACTH(24 hour urine)
● Metyrapone is an inhibitor of 11-beta hydroxylase
Things to consider:
● 24 hour urine free cortisolis themost sensitive and specific screening test
fo r excess cortisol production using HPLC or GC-MS- because plasma cortisol is
affected by diurnal variation
● Urinary free cortisolis theonly portionof cortisol thatpasses through
glomerular filtration
● HPLC-MS- the current reference method for measuring urinary free cortisol
● Insulin Tolerance testis thegold standard for secondary and tertiary
hypocorticolism, confirms borderline response to ACTH stimulation
Standard assessment tests for diagnosing Cushing’s syndrome
Determine Loss of Normal Cortisol Suppression by Dexamethasone
● Use of Dexamethasone
○ Act as exogenous cortisol substitute
○ Suppress the ACTH
■ If the pituitary gland is normal and cortisol if the adrenal gland is normal
● An overnight dexamethasone suppression test is commonly used to screen
patients for autonomous overproduction of cortisol
○
Test
● P
rocedure ofOvernight / Rapid Dexamethasone suppression test
○ 1 mg of dexamethasone is orally given to patient between11pm to 12
midnight
○ Blood is collected the following day8 am to 9 amand urine may be tested for
17 OHCS
○ Normal patient w/o Cushing Syndrome has a cortisol value < 5 ug/dL and 17
OHCS of < 4mg.g creatinine after the test
○ (+) result:all results not suppressed
● Procedure for Low - Dose Dexamethasone suppression test
○ 0.5 mg oral dexamethasone every6 hours for 2 days
○ 24 hour urine and serum samples are collected
○ (+) result:elevated cortisol levels
Congenital Adrenal Hyperplasia Aldosterone
● It results from deficiency of enzymes such as21- hydroxylase deficiency, 11ß M
● ajorelectroregulating hormone
hydroxylase deficiency, 3ß hydroxysteroid dehydrogenase isomerase deficiency ● Most potentmineralocorticoid
(necessary for the secretion of cortisol) ● Asteroid hormonethat helpsregulate water and electrolytes and blood
● This will result todecreased plasma cortisol, increased ACTH and increased levels(Na level in the serum depends almost completely on the interplay
levels of androgens between aldosterone and ADH
● 24 hour urinary free cortisol isnot consistentwithCAH ● Main determinant of renal excretion of potassium
Definitive Tests: ● Acts on renal tubular epithelium to increase retention of Na and Cl, and excretion
● 17-OHP measurement in amniotic fluid of K and H- promotes 1:1 exchange of Na for K or H
● Genotyping cellsfrom chorionic villous sampling-most preferred ● The synthesis of this hormone is primarily controlled by theRAAS
● 18-hydroxysteroid dehydrogenase- an enzyme needed for the synthesis of
aldosterone
● In G-zone
○ The main steroid is the aldosterone
○ Main regulator is the Renin-angiotensin aldosterone system (RAAS/RAS)
○ Major function is blood pressure and potassium (K+) homeostasis
● Syndrome
○ Decrease: Hypoaldosteronism (Distal RTA type IV)
■ Clinical Finding: Hyperkalemia, renal salt wasting
○ Increase:Hyperaldosteronism (aldosteronism)
■ Clinical Finding:HTN, hypokalemia, metabolic alkalosis
Stimulators of aldosterone
● Angiotensin II, ACTH, elevated serum potassium
Inhibitors/Suppressor of Aldosterone
● progesterone and dopamine
● ANP, intracellular calcium, and certain drugs are aldosterone suppressors,
including ketoconazole, ACE inhibitors, nonsteroidal anti-inflammatory drugs, and
heparin
Testosterone
P
● rincipalandrogen hormone- most potent
● Synthesized by the Leydig Cells of the testis of the male, derived from
Progesterone
● Controlled byFSH and LH
Clinical Disorders and Methods
● Function:growth and development of the reproductive system, prostate and
● Clinical Disorders external genitalia
○ Pheochromocytoma ● Levels demonstrate a circardian pattern and peak at (8 am) and fall to their
○ Neuroblastoma lowest level at8 pm
● Methods ● There is gradual reduction of testosterone after age30, with an average decline
○ Chromatography of about 110 ng/dL every decade
○ Radioimmunoassays ● Test for male infertility:
Pheochromocytoma ○ Semen analysis, testosterone, FSH and LH
● Tumors of the adrenal medulla or sympathetic ganglia ● Reference Values: 3.9 – 7.9 ng/mL (serum)
● Commonly seen in 3rd to 5th decades of life (30-50s) ● Transport proteins
● Due tooverproductionof the catecholamine ○ Sex hormone binding globulin (SHBG)- 60%
Signs and symptoms: ○ Albumin- 40%
● Tachycardia, headache, tightness of chest, sweating, hypertension ○ Concentration determines the level of testosterone
Screening Test: Types of Testicular Infertility
● High plasma metanephrines and normetanephrines by HPLC ((plasma-EDTA) ● Pretesticular Infertility (secondary hypogonadism)
Diagnostic Test ○ Due tohypothalamic/pituitarylesions
● High 24 hour urinary excretion of metanephrines and normetanephrines (urine) ○ Testosterone, FSH, LH –Normal/decreased
Pharmacological test ● Testicular Infertility (primary hypogonadism)
1. Clonidine: ○ May be congenital (cryptorchidism, Klinefelter’s syndrome and 5-a-reductase
○ Differentiates pheochromocytoma (not suppressed) to neurogenic deficiency) or acquired (varicocele, tumor, orchitis)
hypertension (50% decreased cathecolamines) ○ Decreasedtestosterone levels andIncreaseFSH and LH
○ Confirmatory: 0.3 mg clonidine → patient → differentiate the borderline ● Post-testicular infertility
result from 1,000-2,000 pg/mL ○ Due to disorders of spermtransport and function
2. Glucagon Stimulation Test ○ Testosterone, FSH, and LH-Normal
○ Used if it is highly suggestive of pheochromocytoma but blood pressure is
Other Disorders of Sexual Development
normal and cathecolamines are modestly elevated: 3 folds increased
1. Testicular feminization Syndrome
Neuroblastoma
○ Most severe form of androgenresistance syndrome, resulting inLackof
● A fatal malignant condition inchildrenresulting toexcessive production of testosterone action in the target tissue
norepinephrine ○ Physical development pursues the female phenotype, with fully developed
● (+) high urinary excretion of (HVA) or VMA or both and dopamine breast and female distribution of fat and hair
● Specimen:24 hour urine and blood (plasma) ○ Noutility or response to administration of exogenous testosterone
Patient Preparation ○ Lab tests:Normallevels of testosterone withelevatedFSH and LH
● The patient should undergoovernight fasting 2. Sertoli-cell only syndrome
● Avoid smoking or drinking coffee at least4 hoursprior to blood collection ○ Characterized by lack ofgerm cells
● The patient is placed in a reclining position in quiet environment and a heparin ○ Men present with small testes, high FSH levels, azoospermia, and normal
lock is inserted intravenously. testosterone level
● After20 to 30 minsblood is collected in apre-chilled EDTA tube ○ Testicular biopsyis the only procedure to confirm this diagnosis
● Plasma concentrations of cathecolamines are affected by body positioning 3. Kallman’s syndrome
and samples must be collected after 30 mins in a stable position decreased ○ A result of an inherited, X-linked recessive trait that manifests as
value when supine hypogonadism during puberty
○ Reclining position Dehydroepiandrosterone
Methods ● The principal androgen formed by the adrenal cortex; weak androgen
● Chromatography ● Derived from the adrenal gland
○ HPLC or GC-MS – (VMA and metanephrines) ● Valuable in the assessment of adrenal cortical hormones
● Radioimmunoassay Estrogen
○ Sensitive screening test for total plasma cathecolamines ● Arises through structural alteration of thetestosteronemolecule
○ > 2000pg/mL of plasma cathecolamines- diagnostic for pheochromocytoma ● Function:promotion of breast development, maturation of external genitalia,
Specimen Consideration deposition of body fat (secondary sexual characteristic of female)
● Catheterization: ● In conjunction withprogesterone, they function in uterine growth and regulation
○ Preferred method of blood collection to eliminate anxiety of venipuncture of menstrual cycle and maintenance of pregnancy
● Urine Preservation ● Deficiency:irregular and incomplete development ofthe endometrium
○ Urine samples with10 mL of 6N HCl(cathecolamines and metabolitesare ● Precursor:acetate, cholesterol, progesterone andtestosterone
rapidly oxidized at higher pH ● Forms:estrone, estradiol and estriol
● 24-hour urine creatinine test ● Estrone and estriol are metabolites of intraovarian and extraglandular conversion
○ To assess the quality of urine collection (0.8 g/day of urine creatinine is A.Estrone
needed to validate thecompleteness of the urinecollection) ● The mostabundantestrogen in postmenopausal women
● To preventcatecholamine oxidation,blood samples must be transported on
B.Estradiol
ice
● Mostpotentestrogen secreted by the ovary, major estrogen
● S ynthesized fromtestosterone, then diffuses out of the thecal cells of the Serotonin (5 hydroxytryptamine)
ovaries of the female
● n amine derived from hydroxylation and decarboxylation of tryptophan
A
● Precursorof both E1 and E3 – serves as negative feedback forFSH ● Synthesized byargentaffin cells, primarily in GI tract
● Used to assessOvarian Function ● Also found in high concentration in pineal gland and CNS
● Transport proteins: Albumin (60%) and SHBG (38%) ● Binds to platelets and released during coagulation
● Free form of E2 is approximately 2% ● Urinary metabolites:5-HIAA
C.Estriol - metabolite of estradiol ● 5-HIAAis a diagnostic marker forcarcinoid tumor
● Estrogen found in thematernal urine ● Test for 5-HIAA:Ehrlich’s Aldehyde
● Major estrogen secreted by the placenta during pregnancy- formation in the Somatostatin
pregnant women is dependent on fetal and placental function
● Also calledGrowth Inhibiting Hormone
● Used to assess the fetoplacental unit (fetoplacental viability), postdate ● Found in the GIT, hypothalamus and delta cells of the pancreas
gestations and intrauterine retardation ● An inhibitor ofGH, Glucagon and insulin
● Used as marker for Down Syndrome Inhibin A
● Preferred specimen:Plasma ● a reproductive system hormone which inhibitsFSH activity
Progesterone Methods
● Produced by the granulose (lutein) cells of the corpus luteum in the female
● Primesecretory productof the ovary
Sample for Hormonal Assay
● Dominant hormone responsible for the luteal phase cycle among females
1. Whole blood: LH and Testosterone
● Single best hormone to determine whetherovulationhas occured
2. Plasma
● Primarily for the evaluation offertilityin female
○ EDTA(ACTH, ADH, PTH)
● Serves to prepare the uterus for pregnancy and the lobules of the breast for
○ Heparin(Cathecolamines, Cortisol, Dopamine, FSH)
lactation
3. Serum
● Deficiency: failure of implantation of the embryo
○ Aldosterone, androstenedione, DHEA, estrogen, FSH, GH, HCG,
● Metabolites: pregnanediols, pregnanediones, pregnanelones
progesterone0
4. Urine: Estriol
Test for Menstrual Cycle Dysfunction Test for femaleInfertility
○ Boric Acid(1 g/dL) preserves estriol and estrogen for 7 days
and Anovulation
○ 10 mL of 6N HClis added to 3-4 L of container (cathecolamines,
Estrogen CG
H
Progesterone PRL vanillylmandelic acid, 5-HIAA)
FSH FT4 Classic Assays
LH TSH ● Bioassays
FSH ● Competitive Protein Binding
LH Immunologic Assays
Estradiol ● Radioimmunoassays
Progesterone ● Immunoradiometric Assays
● Enzyme-linked Immunosorbent Assays
Pancreas ● Enzyme Multiplied Immunosorbent Technique
A digestive gland in thegastrointestinal system
● Fluorescent Techniques
Functions ● Fluorescence Polarization Immunoassay
1. Exocrine:
High Performance Liquid Chromatography
● Responsible for the synthesis of digestive enzymes
●
● Acinus: functional secretory unit
Colorimetry
2. Endocrine
● Responsible for the synthesis ●
● Alpha cells (20-30%) –glucagon Classic Assays
● Beta cells (60-70%) – insulin Bioassay
● Delta cells (2-8%) -somatostatin ● Based on observation of physiologic responses specific for the hormone being
Human Chorionic Gonadotropin (HCG) measures
● Produced by the trophoblast cells of the placenta ● Involvesinjectionof the test materials into animals
● Serves to maintainprogesterone production by the corpus luteum in the early Competitive Binding Protein
pregnancy ● Based on competition for protein-binding sites between a known“tagged/label”
● Can be detected2-3 daysafter ovulation hormone and the unlabeled hormone in the patient’s sample
● Qualitative test for urine samples has detection limit of about 50 mIU/mL ● Measurement of total T4 is based on the specific binding properties of TBG
● Method: Immunometric (sandwich) method Immunologic Assays
Human Placental Lactogen ● widely used to quantify hormones using labeled-antibody withnon-isotopic
● Functionally, structurally and immunologically similar toGrowth Hormone and labels
Prolactin Radioimmunoassays
● Can be measured inUrine, serum, and amniotic fluid ● Is a CBP technique that utilizesradio labeledhormone as the tagged hormone
● Stimulates the development of themammary gland and antisera prepared against the specific hormone as the binding site
● Increases maternal plasma glucose level and promotes positive nitrogen balance Immunoradiometric Assay
● Important in the diagnosis of intrauterine growth retardation ● A radiolabeled substance is attached to the antibody instead of the hormone; an
Gastrin antigen-antibody reaction
● A peptide secreted by the G cells of the antrum of the stomach ELISA
● Released in response tovagalstimulation andFoodin the stomach ● Anenzymeattached to an antibody in which the end product can be measured
● Causes secretion of theHClby parietal cells spectrophotometrically
● Diagnostic marker forZollinger-Ellison syndrome Enzyme Multiplied Immunosorbent Technique (EMIT)
● Major stimulus: presence ofAmino Acid ● Enzyme tags are used; the enzyme is attached to the hormone or drug being
● Increased levels: ZES, Achlorhydria, Chronic RenalFailure tested
Gastric Fluid Acidity ● The rate ofNADHproduced is proportional to the amount of drug tested
● G cells → secrete gastrin ● Requires ofno separationbound and free antigen This method is widely used in
● Parietal cells (Gastrin) → HCl TDM
● Chief cells (produce pepsinogen) → HCl → pepsin Immunometric
BAO BAO/MAO ● ForTSHtest; sensitive test
Basal Acid Output Maximal Acid Output
Fluorescent Techniques
NORMAL 2.5 10%
PERNICIOUS ANEMIA 0 0 Fluorescence Polarization Immunoassay
GASTRIC CARCINOMA 1.0 25% ● flourescein-labeled drug, serum, and antibody are mixed and placed in the light
DUODENAL CANCER 5.0 17% path of a fluorometer.Antibody bound conjugate is inversely proportional to
ZOLLINGER-ELLISON SYNDROME 18.0 72% serum drug concentration
High Performance Liquid Chromatography
● Based on the differentialpartitioningof the compounds between the mobile
phase and stationary phase
Colorimetry
A.Porter-silber method:17-OCHS
B.Zimmermann Reaction
○ measures those steroids with 17-KS
C.Pisano Method
○ For quantifying metanephrines and normetanephrines
D. Kober Reaction
○ For estrogen
○ H2SO4 + hydroquinone
○ (+) reddish brown color
Summary Of Glands And Hormones
Gland Hormones
nterior Pituitary Gland
A H, TSH, ACTH, MSH, LH/ICSH, FSH, PRL
G
Posterior Pituitary Gland ADH, Oxytocin
Thyroid Gland T3, T4, Calcitonin
Parathyroid Gland PTH
Adrenal Gland Epinephrine, norepinephrine, dopamine,
mineralocorticoids, glucocorticoids, adrenal
androgens
ancreas
P Insulin, glucagon, somatostatin
Reproductive Gland Testosterone, estrogens, progesterone
Thymus Gland Thymosin
Pineal Gland Melatonin