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What is the average ejaculate volume?
~2-5ml
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What are the components of Seminal fluid?
- 60% Sminal vesicle fluid ( proteins and flavin)
- 30% Prostatic fluid (AP, PSA)
- 5% epididymis secretions
- 5% bulburethral gland secretions
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The non-cellular portion of semen is known as ________
seminal plasma
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The three gel forming proditens produced by the seminal vesicle are:
- fibronectin
- Semenogelin I
- Semeogelin II
-
Oligospermia
Low sperm count of 20million sperm per ml or less!
-
Azzospermia
lack of sperm in semen aka aspermia
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Two types of Acid Phosphatase
Semianl Acid Phosphatase (SAP) aka Prostatic Acid Phosphatase (PAP) or Prosate Specific Acid Phosphatase (PSAP) or Acid Phosphatase (AP or ACP)
and
Vaginal Acid Phosphatase (VAP)
-
SAP produced in:
Epithelial cells in ducts of prostate gland
-
SAP found:
- High concentration in semen
- Low conc. in blood of healthy males
- Higher levels with prostate cancer
- Male urine
-
SAP structure
dimeric glycoprotein
-
Vaginal AP is present
in vaginal secretions (vaginal drainage) =/
-
Only difference between VAP and SAP
electrocphertic mobility
-
Seminal stains can exhbit AP activity for ____ if stored in a _____
years
cool dry place
Wet or dry seminal stains have been reported to maintain 100% of AP activity after 1 year stored at -20degreees C
-
Dry seminal stains will lose HALF of their AP activity in _____ if stored at 37 deg C
six months
-
Wet seminal stains will lose HALF of their AP acitivity in ________ if stored at 37deg C
2-3 months
-
Prostate Specific Antigen (PSA) also known as:
P-30
-
PSA is present in seminal fluid at concentrations of:
0.5 to 2.0 mg/ml
-
PSA is produced by
Epithelial cells lining the ducts of the prostate gland
-
True or False:
PSA is confirmatory
FALSE!
- some consider it to be however it is not found to be prostate specific
- Found in low levels in: nipple aspirate fluid, breast milk, semen free vaginal swabs, female urine, tumors in female breasts, ovaries, kidneys liver, colon, espohagus, parotid, thyroid and lymphoma
- in blood in healthy males- higher concentration in prostate cancer
-
Approximate total length of a sperm
50-55um
head: 4-5um by 2-3um
-
Abnormally shaped sperm accuont for ______% of total sperm in normal human males
40-60
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Semen Testing:
Screening tests performed first are: _____________
Confirmatory tests are:_____________
Visible stains and latent stains
immunoassays and microscopy
-
Crystalline tests
Presumptive
- The Florence Test- KI and iodine in water
- Positive results w/ vaginal and uterine mucus from dead bodies
- The Barberio Test-picric acide and absolute ethanol solution
- Formed yellow spermine picrate crystals
- Spermine later recognized to be present in many other tissue types
- Puranen's Test- dinitronaphtholsulfonic acid or Napthol Yellow S
- Reacts with spermine
-
Acid Phosphatase Testing
- Most widely used presumptive test
- AP present in high concentrations in semen
- highly stable
- cheap quick and easy!
- BCIP- forms blue precipitate
- Diazo Red- orange/red ppt
- Sodium Thymolphthalein Monophosphate (STMP)-blue color change
- Fast Blue B- (Brentamine Blue) turns dark pink/violet
Tests are qual and semi-quant
-
The Brentamine Reaction (mechanism)
1. a-naphthyl phosphate + Acid phosphatse ---> Free Napthol + Sodium phosphate
2. Free napthol + colorless azo dye ---> Color change
-
In fast blue B the free napthyl reacts with _________to produce a _____ compound
3,3 Dimethoxybenzidine
purple
-
In Diazo red test the free napthyl reacts with the addtion of ______
Fast red RC salt
-
AP Spot the _______ and ________ are already combined to make it a one step test
alpha-napthyl phosphate and Fast blue B
-
Ways to ID PSA
Radial Immunodiffusion: anit P-30 to agarose based gel
Radial diffusion
Rocket Immunoelectrophoresis
Cross over electrophoresis
Immunochromatographic tests
-
Abacus Diagnostics PSA Cards are sensitive to ______
0.004ug/ml or up to 1/1,000,000 semen dilutions
-
_______ were originailly developed as a screenting test for prostate cancer
Abacus Diagnostics PSA cards
-
RSID-Semen is an immunochromatographic test for:
Seminal vesicle-specific Antigen (SVSA)
uses mouse monoclonal antibodies specific for human semenologin
-
Seminal vesicle-Specific antigen (SVSA) is found in _______ and _______
Semenogelin I and II
-
____1___ is on the test pad
___2____ are immobilized on the T line
The C line has _____3_____
1-monoclonal mouse antibodies
2- additional mouse antibodies immobilized
3- immobilized anti-mouse IGg acts as control
-
ELISA is sensitive to:
Less than 10ng/ml
-
_____ utilizes a solid-phase double antibody sandwich assay
Enzyme Linked immunosorbent assay (ELISA)
-
ELISA uses:
polyclonal or monoclonal anti-P30
-
Dried semen stains may fluoresce under
- Short wave and long wave UV
- an argon laser
- wavelenghts between 450-495nm with an orange barrier filter (interference filter of ~530nm)
-
How ALS works:
The emitted light has a relatively ____ intensity that is overpowered by the ______light
To observe fluoresecne ______ must be filtered out
-
The single best confirmation of semen:
Microscopic analysis!
-
A _____ is useful in identifying intact sperm on unstained smear slides
Phase contrast microscope
-
Nuclear fast red will stain _____ red
Sperm heads
-
Picroindigocarmine will stain____ and ____ green
Sperm tails and cytoplasm of e-cells
-
In Xmas tree stain: ______ may also stain red
Bacteria, yeast and WBC
-
Hemotoxlyin ane eosin stains:
Purple and pink
- Heads:purple
- Tails: pink
- Ecell cytoplasm: pink
- Ecell nuclie: purple
-
Positive AP or other preliminary color test:
Analysis of_____ gave presumptive chemical indication for the presence of AP, a component of semen
-
Positive ALS
Numerous fluorescing stains were observed on ____ using an ALS
-
Positive AP and Positive P30 with Negative microscopic
Components of semen were identified on ______, however microscopic examination failed to demonstrate presence of spermatozoa
-
Positive AP, Negative microscopeic, negative P30
No semen was identified on____
-
Negative AP
Analysis of ____failed to demosntrate presence of semen
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