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Saliva
For Final Exam
22
Biology
Undergraduate 3
04/30/2013

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Term
How much saliva do the human salivary glands produce per day?
Definition
1.0-1.5 L/day
Term
What is saliva?
Definition
A slightly alkaline secretion, comprised of water (mostly), mucus, proteins, salts, and enzymes
Term
Tests for saliva
Definition
NO test is specific for saliva
Idntification of amylase indicates saliva
Term
Amylase
Definition
Enzyme that cleaves polysaccharidse that are composed of D-glucose units at the alpha1-4 linkages. AKA cleaves starch.
Digests carbohydrates in the oral cavity
Term
Starch
Definition
Contains two glucose polymers: amylose and amylopectin
Term
Amylose
Definition
Long, unbranced glucose connected at the alpha1-4
Term
Amylopectin
Definition
Branched alpha1-6 connections and linear alpha1-4
Term
Two Types of Amylases
Definition
beta-amylase in plant and bacterial sources
alpha-amylase in human, human saliva (HSA), and human pancreatic (HPA)
Term
Human alpha-amylases: Two isoforms
Definition
Differ in amino acid sequence
Human alpha-amylsase (HSA)
Human pancreatic amylase (HPA)
Term
Human Alpha-Amylase (HSA)
Definition
From saliva
Inactivated by stomach acids
AMY1 locus
Term
Human Pancreatic Alpha-Amylase (HPA)
Definition
From pancreas
Inactivated in the intestines with some activiy remaining in the feces.
AMY2 locus
Term
Where is amylase found?
Definition
Semen, tears, milk, persperation, vaginal secretions, serum, and urine
Term
Presumptive Assays
Definition
Visual examination with ALS or microscope
Starch-iodine assays
Colorimetric Assays
Term
Visual Examination: ALS or Microscope
Definition
Can see mucous cells to test if it is saliva
ALS: Fluorescence of saliva is less intense than that of semen
Microscopic: stain for buccal epithelial cells
Term
Starch-iodine Assays
Definition
Iodine used to test for starch
NOT specific because it is technically for STARCH
Starch appears blue, amylopectin appears reddish-purple in the presence of iodine
Albumin and other proteins produce a false positive
Term
Colirmetric Assays
Definition
Dye-labeled amylase substrates are employed
Phadebas Test
Press Test
Term
Phadebas Test
Definition
Clinical diagnosis & forensic use
Procion Red Amylopectin (PRA) or phadebas reagent:
-starch is linked to the dye forming insoluble complex
-dye becomes sluble, creating a colored product measured by a spectrophotometer
-done via paper or tube method
-may not detect low levels
Term
Press Test
Definition
-filters are sprayed with Phadebas solution and allowed to air dry
-paper soaked with dried Phadebas solution is pressed over the area to be tested
-initial time, single minutes for the first 10 minutes, and 5 minutes for up to 40 minutes total = intervals to note observations
Appears light blue on surface of paper
Term
Confirmatory Assays
Definition
Immunochromatographic Assays - RSID-Saliva
ELISA - monoclonal HSA antibody
RNA-based Assays
-based on certain genes specifically expressed in certain types of cells
-based on mRNA & htRNA
Term
Confirmatory Assays: Immunochromatographic Assays
Definition
RSID-Saliva
Sensititivy: 1 microL saliva necessary
specificity: no cross reaction with animal saliva, earwax, etc. most body fluids are okay.
utilizes 2 anti-salivary alhpa-amylase monocolonal antibodies in a lateral flow immunochromatographic strip test formation to detect the presence of salivary amylase rather than the activitiy of the enzyme
No high dose Hook effect
Term
Confirmatory Assays: ELISA
Definition
monoclonal HSA (human salivary alpha-amylase) antibody
Only cross-reactivity for chimpanzee, mouse, rat, and eel
Did not work on old stains
Specific and can be used for quantification
Term
Confirmatory Assays: RNA-based Assays
Definition
mRNA that is expressed only in the oral cavity
Uses reverse transcriptase - PCR (RT-PCR) to detect levels of mRNA
HTN3 encodes histain 3 - a protein involved in non-immune response host defense
STATH encodes statherin - inhibits precipitation of calcium salts in the salivary glands & oral cavity
Highly specific and can be automated, BUT RNA is unstable due to degradation
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