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PATH 406 > Reproductive Endocrinology > Flashcards

Reproductive Endocrinology Flashcards

1
Q
  1. What is FSH produced by?
    a. What is the molecular weight of FSH?
    b. What kind of dimer is FSH?
    c. How many subunits does FSH have? State the length of the subunit.
    d. Immunoassays are specific for which subunit?
A
  1. Pituitary gland
    a. 30000 Da
    b. Heterodimer
    c. 2 subunits, alpha and beta.
    Alpha (116 aa) - 14000 Da
    Beta (129 aa)
    d. Beta dimer
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2
Q

What does FSH act on and what does it do?

A

Ovarian Follicles
1. FSH stimulates follicular development and production of estradiol and other estrogens during the follicular phase of the menstrual cycle.
2. Acts synergistically with LH to cause ovulation

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3
Q

How do we detect FSH?

A

Siemens Centaur
Siemens Immulite

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4
Q

What kind of procedure is the Siemens Centaur? How does the Siemens Centaur Methodology work?

A

This FSH procedure is a 2-site chemiluminescent immunoassay.
1. FSH in patient’s serum binds to an acridinium labeled mouse monoclonal Ab
2. A second Ab specific for FSH attached to magnetic particles captures the complex
3. Magnetic particles are separated
4. Acridinium ester remaining on solid phase reacts chemically to release light
Amount of FSH is directly proportional to RLUs (Relative light units)

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5
Q

How does the Siemens Immulite Methodology works?

A
  1. Latex beads coated with monoclonal mouse anti-FSH are added
  2. After incubation, a second monoclonal mouse anti-FSH conjugated with bovine ALP is added
  3. Latex beads allow separation
  4. ALP activity can be measured by adding a substrate for ALP that is chemiluminescent when it reacts

Amount of FSH directly proportional to signal

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6
Q

What are the reference ranges for FSH?

A

Reference Intervals for pubescent females by Tanner stage:
I - 0.4-6.5 IU/L
II - 1.0-8.4 IU/L
III - 1.0-9.5 IU/L
IV/V - 0.6-9.4 IU/L

Reference Intervals for Adult Females
Follicular: 3.5-12.5 IU/L
Mid-cycle: 4.7-12.5 IU/L
Luteal: 1.7-7.7 IU/L
Post-menopausal: 25.8-134.8 IU/L

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7
Q
  1. What is LH?
    a. What is the molecular weight of LH?
    b. What is the structure of LH?
    c. What are the lengths of the subunits and the mass?
    d. Where is the alpha and beta subunit found?
    e. Which subunit does IA detect?
A
  1. Luteinizing hormone
    a. 32000 Da
    b. Heterodimer, alpha and beta subunit
    c. Alpha (116 aa) 14000 Da,
    Beta (141 aa)
    d. Alpha common in gonadotrophins and TSH while beta specific to LH because of its aa sequence and carbohydrate content
    e. B
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8
Q

What is a way to measure LH?

A

Two-site immunometric
- Spectrophotometric
- Chemiluminescent

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9
Q

What is the reference range for LH?

A

RI for pubescent females:
I - <2.9
II - <8
III - <23.1
IV/V - <25.4

RI for adult females
Follicular - 2.4-12.6
Mid-cycle - 14-95.6
Luteal - 1-11.4
Post menopausal - 7.7-58.5

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10
Q
  1. What kind of steroids are estrogens?
  2. What are the 3 important estrogens?
  3. What are the characteristics of the 3 types of estrogens?
A
  1. C-18 steroids
  2. Estradiol (E2), Estrone (E1), Estriol (E3)
  3. E2 - most biologically active
    E1 - relatively inactive
    E3 - predominates in pregnancy, derives from fetoplacental unit
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11
Q

What are the actions of E2?

A
  1. Development and maintenance of female sexual characteristics
  2. Endometrial proliferation
  3. Breast ductal manipulation
  4. Decrease bone resorption
  5. Increase sex hormone binding globulin (SHBG), cortisol binding globulin (CBG), and thyroxine binding globulin (TBG)
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12
Q

How is E2 transported?

A
  1. E2 is a non-polar molecule that is required to be transported in aqueous phase
  2. Main transporter proteins are SHBG and albumin
  3. Albumin is a high capacity, low affinity binder carrying 20-30%
  4. SHBG is a low capacity, high affinity binder carrying 70-80%
  5. About 2-3% of E2 is free and biologically active
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13
Q

What does the term bioavailable estrogen refer to?

A

The sum of the free and the albumin bound estrogen

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14
Q

What do we need to do before measuring E2?

A
  1. Use of competitive assay is required because E2 is small
  2. Due to the binding of SHBG and albumin, E2 needs to be freed by precipitation with the addition of 8-napthalene-sulfonic acid
  3. Dihydrotestosterone can also be used because it has greater affinity for SHBG
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15
Q

What are the methods used to measure E2? How do we interpret the results? What is the reference method?

A

RIA
Non-isotopic with chemiluminsecent detection

Signal is inversely proportional with the analyte concentration

GC-MS is the reference method

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16
Q

When does E3 become the major plasma estrogen?

A

During the third trimester of pregnancy

17
Q

How is E3 produced?

A

Cholesterol to DHEAS to 16-OH-DHEAS to 16-OH-AD to E3

Fetal adrenal to fetal liver to placenta to placenta

18
Q

What happens to E3 in the maternal serum?

A

Conjugated to either sulfate or glucuronic acid and excreted out through the maternal urine

Unconjugated E3 (uE3) is not soluble and therefore is strongly bound to SHBG

19
Q

What is uE3 measured by?

A

Autodelfia method - competitive, time-resolved fluorescence

20
Q

What type of steroid is progesterone? What is the main distinctive organic feature? What is progesterone primarily transported by?

A

C21 steroids
alphabeta unsaturated ketone at positions 3, 4 and 5
CBG, 2-10% free

21
Q

What is the reference method for progesterone?

A

GC-MS

22
Q

What is primary amenorrhea?

A

Failure to establish spontaneous periodic menses by 16 yo

23
Q

What kind of steroids are testosterones?

A

C-19 steroids

24
Q

What is the main androgen produced by the Leydig cells of the testicle?

A

Testosterone

25
Q

What are the two most important androgens?

A

Testosterone and dihydrotestosterone
T and DHT

26
Q

What other hormones do the testicles produce beside T and DHT?

A

Dihydroepiandrosterone (DHEA)
Androstendione

27
Q

What hormones does the adrenal produce?

A

DHEA
DHEA-S
Androstendione
Androstendiol

28
Q

What are the actions of testosterone?

A
  1. Masculinization of genital tract
  2. Development and maintenance of secondary sexual characteristics
  3. Muscle bulk, bone mass, libido, sexual performance
29
Q

How is testosterone transported?

A

Main transport proteins are SHBG and albumin
1. 40-60% SHBG bound
2. 30-50% albumin bound
3. 2-3% free

30
Q

What is the reference method for testosterone?

A

GC-MS

31
Q

What does the term bioavailable testosterone refer to? How to calculate?

A

Sum of free + albumin-bound testosterone
Vermeulen equation

32
Q

What does free testosterone refer to? How to quantify free testosterone?

A

Sum of freely dissolved testosterone
1. Equilibrium dialysis and Immunoassay or LC-MS/MS

33
Q

When is DHEAS elevated?

A
  1. PCOS
  2. Congenital adrenal hyperplasia (CAH)
  3. Tumors of the adrenal cortex
34
Q

Is protein binding an issue with DHEAS?

A

No because it is water soluble

PATH 406 (24 decks)

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