Hormonal Assays and Interpretation

Hormonal Assays and Interpretation

Hormonal Assays and Interpretation

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Assay Principles - Hormones Under Scope

  • Immunoassays: Core of hormone measurement.
    • Competitive: Labeled & unlabeled hormone compete for limited antibody sites. Signal $\propto$ 1/[Analyte].
    • Non-competitive (Sandwich): Analyte captured between two antibodies. Signal $\propto$ [Analyte].
  • Key Techniques:
    • ELISA: Enzyme-linked, colorimetric/fluorometric signal.
    • RIA: Radiolabeled antigen ($^{125}$I), highly sensitive.
    • CLIA: Chemiluminescent signal, high sensitivity & wide range.
    • LC-MS/MS: Gold standard for steroids; high specificity & multiplexing.
  • Hormones Covered: Thyroid (TSH, T3, T4), PTH, Cortisol, Aldosterone, GH, ACTH, Prolactin, Insulin, Estrogen, Testosterone, hCG. Direct vs. Indirect Competitive Immunoassay

⭐ Free hormone fractions (e.g., fT4, fTestosterone) often better reflect biological activity than total hormone levels, as they are unbound to proteins.

Pituitary & Thyroid - Master Regulators

  • Pituitary Gland:
    • Anterior Lobe: Secretes TSH, ACTH, GH, PRL, FSH, LH. 📌 Mnemonic: FLAT PiG.
      • TSH (Thyrotropin): Stimulates thyroid hormone (T3, T4) synthesis/release. Regulated by hypothalamic TRH (+) and thyroid hormone (-).
    • Posterior Lobe: Stores & releases ADH, Oxytocin (synthesized in hypothalamus).
  • Thyroid Gland:
    • Hormones: Thyroxine (T4), Triiodothyronine (T3), Calcitonin.
    • T4 is prohormone; converted to T3 (more potent) in periphery by deiodinases.
    • Binding proteins: TBG (Thyroxine-binding globulin) carries most T4/T3.
  • Hypothalamic-Pituitary-Thyroid (HPT) Axis:
  • Thyroid Function Tests (TFTs) Interpretation:
    • TSH: Best initial screening. Normal: 0.4-4.0 mIU/L.
      • Primary Hypothyroidism: ↑ TSH, ↓ Free T4.
      • Primary Hyperthyroidism: ↓ TSH, ↑ Free T4.
      • Subclinical Hypothyroidism: ↑ TSH, Normal Free T4.
    • Anti-TPO Abs: Hashimoto's. TSH-Receptor Abs (TSI): Graves' disease.

⭐ TSH is the most sensitive indicator of primary thyroid dysfunction; changes in TSH often precede changes in T4/T3 levels.

Adrenal & Gonadal - Stress & Sex Hormones

  • Adrenal Cortex:
    • Cortisol (Glucocorticoid): Stress response, ↑glucose. Reg: CRH-ACTH-Cortisol.
      • Tests: Dexamethasone suppression, ACTH stimulation, 24hr Urinary Free Cortisol (UFC).
      • Disorders: Cushing's syndrome (↑cortisol), Addison's disease (↓cortisol).
    • Aldosterone (Mineralocorticoid): Na⁺/H₂O reabsorption, K⁺ excretion, BP control. Reg: RAAS, K⁺.
      • Tests: Aldosterone:Renin Ratio (ARR). Conn's syndrome (↑aldosterone).
    • Adrenal Androgens (DHEA, DHEAS): Precursors to sex hormones.
      • Test: 17-hydroxyprogesterone (17-OHP) for Congenital Adrenal Hyperplasia (CAH).
  • Adrenal Medulla:
    • Catecholamines (Epinephrine, Norepinephrine): Fight-or-flight.
      • Test: Plasma/Urinary Metanephrines, Vanillylmandelic Acid (VMA) for Pheochromocytoma.
  • Gonadal Hormones:
    • Testosterone (Androgen): Male 2° sexual characteristics, spermatogenesis.
      • Tests: Total/Free Testosterone, LH, FSH.
    • Estrogen (Estradiol) & Progesterone: Female 2° sexual characteristics, menstrual cycle, pregnancy.
      • Tests: Estradiol, Progesterone, LH, FSH. Polycystic Ovary Syndrome (PCOS) common.

Adrenal and Gonadal Hormone Synthesis Pathways

⭐ 21-Hydroxylase deficiency is the most common CAH cause: ↓cortisol, ↓aldosterone, ↑androgens. Presents with salt wasting & virilization (females).

Pancreas & Calcium - Metabolic Managers

  • Pancreatic Hormones:
    • Insulin (β-cells): Anabolic. ↓ Blood glucose. C-peptide reflects endogenous insulin.
    • Glucagon (α-cells): Catabolic. ↑ Blood glucose, glycogenolysis, gluconeogenesis.
    • Somatostatin (δ-cells): Universal inhibitor (insulin, glucagon, GH).
  • Calcium Homeostasis:
    • PTH: ↑ Serum $Ca^{2+}$, ↓ Serum $PO_4^{3-}$ (📌 Phosphate Trashing Hormone). Stimulates osteoclasts, renal $Ca^{2+}$ reabsorption, Vit D activation.
    • Vitamin D ($1,25(OH)_2D_3$): ↑ Intestinal $Ca^{2+}$ & $PO_4^{3-}$ absorption.
    • Calcitonin (Thyroid C-cells): Weakly ↓ Serum $Ca^{2+}$ by inhibiting osteoclasts.

⭐ Primary hyperparathyroidism classically presents with high PTH, high serum calcium, low serum phosphate, and increased urinary cAMP. Calcium Homeostasis: PTH, Calcitonin, and Calcitriol Actions

High‑Yield Points - ⚡ Biggest Takeaways

  • Immunoassays (ELISA, RIA, CLIA) are mainstay for hormone quantification.
  • Dynamic function tests (e.g., Dexamethasone suppression) assess endocrine reserve/autonomy.
  • Free hormone levels (e.g., FT4) better reflect bioactivity than total hormone levels.
  • Pulsatile secretion (LH) & diurnal variation (cortisol) demand timed/pooled sampling.
  • Assay interferences (e.g., HAMA) can cause falsely high or low results.
  • Interpret results with clinical context, considering age, sex, pregnancy, and medications.

Practice Questions: Hormonal Assays and Interpretation

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Which hormone is released when serum calcium levels decrease?

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Flashcards: Hormonal Assays and Interpretation

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_____ is diagnosed by increased serum and urine levels of catecholamines, or their breakdown products metanephrines and vanillylmandelic acid (VMA)

TAP TO REVEAL ANSWER

_____ is diagnosed by increased serum and urine levels of catecholamines, or their breakdown products metanephrines and vanillylmandelic acid (VMA)

Pheochromocytoma

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