Clinical Chem Cls 2213 Unit 8

Front 1

enzyme lacking PKU

Back 1

phenylalanine hydroxylase is missing which converts phenylalanine into tyrosine
-at birth screening test is Guthrie test

Front 2

enzymes lacking in tyrosinosis

Back 2

-tyrosine transaminase which converts tyrosine into p-hydroxy phenylpyruvic acid
- or p-hydroxy-phenyl-pyruvate hydroxylase which converts the p-hydroxy phenylpyruvic Acid into homogentistic acid

Front 3

enzyme lacking in alkaptonuria

Back 3

homogentistic acid oxidase is missing which converts homogentistic acid into maleylaceto-acetic acid

Front 4

enzyme lacking in maple syrup disease

Back 4

branched chain keto acid decarboxylase deficiency

Front 5

symptoms and urine findings in PKU

Back 5

siezures, mental retardation
urine- increased phenylalanine

Front 6

symptoms and urine findings in cystinuria

Back 6

-urinary calculi
-Urine- elevated levels of cystine, cysteine, lysine, arginine, and ornithine

Front 7

symptoms and urine findings in tyrosinosis

Back 7

enlarged liver and spleen. severe liver damage, may be fatal
Urine- elevated levels of tyrosine and DOPA

Front 8

symptoms and urine findings in alkaptonuria

Back 8

black pigmentation of ears and other cartilage; arthritis
Urine- homogenitisic acid very elevated, urine darkens within 24 hrs as HGA converts spontaneously to melanin

Front 9

symptoms and urine findings in maple syrup disease

Back 9

severe mental retardation, early infant death, symptoms appear during first week of life
Urine- odor of maple syrup, elevated leucine, isoleucine, valine, and keto acids

Front 10

Tests for PKU

Back 10

FeCl3= Green
DNPH Test= positive

Front 11

Tests for Cystinuria

Back 11

Cyanide-Nitroprusside test= pos

Front 12

Tests for Tyrosinosis

Back 12

DNPH= Pos
FeCl3= quick fading green

Front 13

Tests for alkaptonuria

Back 13

FeCl3= transient blue
Clinitest pos

Front 14

Tests for maple syrup disease

Back 14

DNPH= pos
FeCl3= blue

Front 15

Enzyme absent in histidemia

Back 15

Histidase

Front 16

Enzyme problem in non-ketotic hyperglycemia

Back 16

Block in glycine cleavage enzyme system

Front 17

Enzyme deficiency in Methylmalonic acidemia

Back 17

Methylmalonyl CoA mutase

Front 18

Symptoms and urine findings in Histidinemia

Back 18

sometimes normal, sometimes neurological symptoms like retardation, or speech defects
Urine- Imidazole; pyruvic acid and other histidine metabolites

Front 19

Symptoms and urine findings in Non-ketotic hyperglycemia

Back 19

Siezures, hypotonia, no ketosis. If severe mental retardation, fatal within 2 yrs
Urine- excessive Glycine (also in CSF)

Front 20

Symptoms and urine findings in Methylmalonic acidemia

Back 20

Metabolic ketoacidosis, developmental retardation
Urine- Excessive Glycine; methylmalonic acid (also in CSF); ketonuria

Front 21

Amino acid composition

Back 21

- Amine group
- Carboxy group
- Hydogen
- H group (another thingy that makes it unique)

Front 22

What are poly-peptides

Back 22

many AA's bonded together, but total molecular weight is less than 10,000. More than 10,000 is a protein

Front 23

what is primary, secondary, tertiary, and quaternary structure

Back 23

Primary- sequence of AA
Secondary- coiling of AA
Tertiary- coiling causes one part to hydrogen/ionically bond with another part
Quaternary- Molecule consists of tow or more different proteins

Front 24

Albumins

Back 24

true proteins- consist entirely of AA chains

Front 25

Globulins

Back 25

AA chains forming true proteins but contain non-AA constituents. Not as soluble as albumins

Front 26

Types of globulins

Back 26

AA chains plus;
CHO=glycoproteins
Lipids= lipoproteins
metals= metaloproteins
Funky amine-CHO= mucoproteins

Front 27

Explain TLC

Back 27

Thin Layer Chromatography
- add amino acid controls and sample to paper
- place in solvent to get separation of individual analytes
- take the Rf ratios for ID

Front 28

Explain liquid chromatography

Back 28

use particles or gel beads
-Solvent 1= stationary phase
- Solvent 2= mobile phase
- seperation based on solubility in solvent #1 vs. #2

Front 29

Concept of ion-exchange chromatography

Back 29

-mixture of analytes at top
- charged resin
- separation of individual analytes by charge

Front 30

HPLC

Back 30

High Pressure Liquid Chromatography
- like liquid-chromatography but under extremely high pressure
- elution time is very fast
- works for volatile and non-volatile analytes

Front 31

Reagents and principles with Ninhydrin

Back 31

-Stains all AA's some hue of pink to purple
- if it has an Alpha Carbon with an amine group and a carboxyl group attached, it will stain with Ninhydrin
- some AA's are distinguishable by color- Aspartic acid= purple

Front 32

Reagents and principles with Bromcresol Green

Back 32

-Most commonly used serum albumin reagent
- Albumin will bind BCG dye and create a Green complex
- will not bind any globulin proteins
- Bromcresol purple is a similar reagent

Front 33

Reagents and principles with Biuret reagent

Back 33

Copper sulfate in NaOH, KI, and gobs of Na, K-trartrate
- Reagent is only acceptable for Serum protein assays. Linear range doesn't go below 1.0 to 2.0 g/dL
-Will react with tri-peptides and larger
-Will not react with single AA's

Front 34

Reagents and principles with refractive index

Back 34

-Proteins influence the refractive index of light
- Special protein ocular can be obtained and used with a refractometer
- Slow manual method (not a RED HOT method)

Front 35

Reagents and principles with Turbidimetric Protein Assays

Back 35

Protein in small amounts such as in urine and CSF can be quantitated photometrically by precipitating them.
- the more cloudiness the more protein
- linear range is 10-150 mg/dL
- Method uses TRICHLOROACETIC ACID (TCA) and SULFOSALYCYLIC ACID

Front 36

Reagents and principles with Coomassie Blue

Back 36

-Reagent comes in concentrated form and is RED
- Our MOST SENSITIVE method of measuring extremely low levels of protein, especially in urine
-linear range can go as low as 5-3 mg/dL

Front 37

Reference range for serum or plasma albumin

Back 37

3.5-5.2 g/dL

Front 38

Reference range for serum or plasma total protein

Back 38

6.0-8.2 g/dL

Front 39

Reference range for serum or plasma A/G ratio

Back 39

1.1-1.8

Front 40

Reference range for CSF protein

Back 40

15-40 mg/dL

Front 41

Reference range for CSF glucose

Back 41

40-70 mg/dL or 2/3 of current blood glucose level

Front 42

Reference range for urine protein

Back 42

50-150 mg/day (<10mg/dL random collection)

Front 43

Causes for elevated protein in CSF

Back 43

Bacterial meningitis
Viral (normal glucose)
Chemical (normal glucose)
Encephalitis
MS
Sub-arachnoid hemorrhage

Front 44

Causes for elevated Urinary proteins

Back 44

Acute glomerulonephritis
Acute pyelonephritis
Nephrotic Syndrome (albumin must be <3.0 gm/dL and tot. PRO <5.0 gm/dL. Oval fat bodies in urine, elevated serum cholesterol)
Multiple Myeloma- due to increased production of LIGHT CHAIN antibody components that are small enough to be excreted

Front 45

Causes for elevated protein in Microalbuminuria

Back 45

High blood pressure causes excessive albumin excretion.
Normal protein excretion is strictly Tamm-Horsfall protein, not albumin or the other globulins

Front 46

list the 5 major serum fractions obtained through Cellulose Acetate Electrophoresis

Back 46

Albumin- 57% or 4.56 g/dL
Alpha-1- 3.5% or 0.28 g/dL
Alpha-2- 11.5% or 0.92 g/dL
Beta- 12.5% or 1.00 g/dL
Gamma- 15.5% or 1.24 g/dL

Front 47

Buffer pH in CAE

Back 47

8.6-8.8

Front 48

Direction of protein migration in CAE

Back 48

Proteins migrate toward the anode

Front 49

Stain for CAE

Back 49

Ponceau-S Red

Front 50

What makes up the Alpha-1 group in CAE

Back 50

Alpha-1 Anti-trypsin(90%)
Alpha lipoproteins
Alpha-1 glycoprotein

Front 51

What makes up the Alpha-2 group in CAE

Back 51

Ceruloplasmin
Haptoglobin
Alpha-2 Macroglobulin
Pre-beta lipoproteins

Front 52

What makes up the Beta group in CAE
Chunk Coerced The Fertellis By Being Honest

Back 52

C-Reactive Protein
Complement c-3 and c-4
Transferrin
Fibrinogen
Beta lipoproteins
Beta-2 microglobulins
Hemopexin

Front 53

What makes up the gamma group in CAE

Back 53

IgG
IgM
IgA
IgE

Front 54

What is Wick-Flow

Back 54

Evaporation due to heat
minimized by using otter pops

Front 55

What is the advantage of Constant current as compared to constant-voltage

Back 55

Constant current minimizes heat production where as constant voltage tends to create excessive hear

Front 56

Equation for electrophoresis migration rate

Back 56

u=netcharge/K x Size x Buffer viscosity

Front 57

what problems can occur with improper buffer

Back 57

If pH is wrong- afffects charge and direction of migration
If ionic strength wrong- affects migration and seperation

Front 58

what problems can occur with unsatisfactory staining

Back 58

if stain washes off during the 5% acetic acid washes, the stain is too old
- if it fails to stain too little sample was applied

Front 59

what problems can occur with incorrect amount of sample applied

Back 59

If sample is over applied- scanned albumin will be LOW, and globulin will be HIGH
- if wample was under applied- albumin will be HIGH and globulin will be LOW

Front 60

what problems can occur with discontinuities in sample application

Back 60

if applicator wire is bent- bow tie patterns or won't pick up correct amount of sample

Front 61

what problems can sause distortion of protein zones

Back 61

-Dirt on membrane
- strip didn't hydrate evenly
- Not sufficient blotting prior to applying sample
- too much blotting dries out membrane
- wiggling during sample application

Front 62

what problems can cause artifacts

Back 62

-Buffer contaminated with bacteria
- bent applicator wires
- membrane allowed to dry-out
- membrane blotted too much

Front 63

what problems can cause the proteins to not move

Back 63

-Short in electrophoresis cell
- Jayson didn't turn on power source
- No wicks in chamber
- Cellulose acetate side is not touching wicks
- Wrong buffer- pH is at proteins pI

Front 64

special sample prep steps for CSF and urine

Back 64

1- centrifuge to remove swimmies and floaties, and other artifacts
2- concentrate the sample using osmosis chambers

Front 65

Monoclonal spike

Back 65

A single protein is in abundance. typical of multiple myeloma

Front 66

Polyclonal spike

Back 66

A broad spike that representsa bunch of proteins being produced at the same time, like a bunch of different antibodies

Front 67

Bence jones protein

Back 67

-Found in multiple myeloma
-Light chain protein
-Will heat coagulate at 56c then will redissolve at 90-100c

Front 68

Ceruloplasmin disorders

Back 68

- Carries copper
- decreased in liver disease and especially in Wilson's disease
- A lack causes copper deposits in tissues causing a brown halo in the iris

Front 69

Alpha 1 anti-trypsin alleles

Back 69

M, S, and Z
-MM is normal
-ZZ develops premature emphysema
- liver production of AAT increases during inflammation

Front 70

Transferrin stuff

Back 70

-NOT an acute phase protein
- drops during inflammation
- increases in iron deficiency anemia

Front 71

other name for pre-albumin

Back 71

Transthyretin
- increasing levels are a sign of good prognosis
- sort of a wellnes marker

Front 72

How does CAE help diagnose MS

Back 72

the presence of oligoclonal bands in the gamma region indicate MS autoimmune antibodies

Front 73

Details on Multiple Myeloma

Back 73

Multiple clones of malignant plasma cells often causing "spontaneous Fracture"
- production of gobs of heavy and light chain proteins that are never made into antibodies
- Heavy chains back up in the blood causing hyperviscocity syndrome resulting in rouleoux
- If light chain is Kappa then good prognosis, if Lambda then poorer prognosis
- 85% of diseases are heavy and light chain
- 15% are light chain only-these have a worse prognosis

Front 74

Alpha 1 fetal protein

Back 74

used as an adult serum marker for
- Primary liver cancer
- Cirrhosis
- Hepatitis B and C
- Testicular carcinomas

Front 75

Benefits of Agarose Gel

Back 75

Gives better resolution that cellulose acetate

Front 76

Benefits of Polyacrylamide gels

Back 76

Extremely high resolution for things like HIV antibodies and antigens
-media is colored so can't be scanned

Front 77

Benefits of immunoelectrophoresis

Back 77

-possible to isolate each individual protein

Front 78

Benefits of immuno-fixation electrophoresis

Back 78

anti-sera is poured on top of gel and precipitation rxn occurs within an hour. Each protein has it's own precipitation band

Front 79

Benefits of immunonepholometry

Back 79

a way to quantitate individual proteins like AAT