3. Phenylalanine-Tyrosine Disorders
β
β
Many of most frequently requested special urinalysis
assoc. w/ this metabolic pathway
Major inherited disorders:
PKU
β Tyrosyluria
β Alkaptonuria
Metabolic Defects: Melanoma
β
β
4. Phenylketonuria
β
Most well known aminoaciduria
β
1/10k-20k births
β
Can cause:
Severe mental retardation if undetected
β Fair complexions due to decreased prod of
tyrosine & melanin
Identified in Norway by Ivan Folling (1934)
β
β
β
β
β
Peculiar mousy odour of urine of mentally retarded
child
High levels of keto acids, inlcuding
phenylpyruvate
Conversion of phenylalanine to tyrosine is disrupted
5. β
β
β
Absence of gene needed to prod.
Phenylalanine hydroxylase
Autosomal recessive trait: normal
heterozygous carrier
Newborn screening test: (+) PKU = eliminate
phenylalanine in diet (major constituent in milk)
= prevent excessive buildup in serum = avoid
mental retardation
β
Alternate pathways develop as child matures
β
Aspartame: high levels of phenylalanine
6. β
Blood Test
β
β
β
β
β
Initial screening
done b/f being discharged from hospital
Can be detected 4hrs after birth if cutoff is
lowered to 2mg/dL
May need to repeat test in early visit to
pediatrician
Slower rise in girls- more prone to escape
detection during early tests
7. β
Microbial Inhibition Assay β Guthrie
β
β
Most well-known blood test for PKU
P: phenylalanine counteracts action of beta-2thienylalanine, an inhibitor of B. subtilis
1. Blood from heelstick absorbed into filter
paper circles
2. Paper disks placed on CM streaked w/
B. Subtilis
(+) growth around paper disk
(-) no growth
8. β
Urine Test
β
β
β
β
Appears after 2-6 weeks in urine
Follow up procedure in questionable diagnostic
cases
Screening test to ensure proper dietary control
Monitoring dietary intake of pregnant women
lacking phenylalanine hydroxylase
9. β
Ferric Chloride
β
β
β
β
Urine tube test testing for phenylpyrvic acid
Nonspecific: reacts with other amino acids &
medications
False (+) produced by certain brands of
disposable diapers
Permanent blue-green colour
1. Place 1mL of urine in tube
2. Slowly add 5 drops of 10% ferric chloride
3. Observe colour
10. Tyrosyluria
β
Tyrosinemia produces urinary overflow
β
Inherited/Metabolic
β
Since 2 reactions are directly involved in tyrosin
metabolism, urine may have:
β
β
Excess tyrosine
Excess degraded products:
β P-hydroxyphenylpyruvic acid
β P-hydroxyphenyllactic acid
11. β
Transitory Tyrosinemia
β
β
β
β
Most frequently seen in premature infants
Caused by underdevelopment of liver fxn req.
to prod. enzymes needed for complete
metabolism
Rarely causes permanent damage
Difference in urinary test w/ PKU
β Green colour fades away quickly
12. β
Acquired liver Disease
β
Resembles transitory newborn tyrosyluria
β
Leucine & Tyrosine crystals seen
β
Hereditary Disorders
β
β
β
β
β
Absence of enzymes needed in metabolic pathway
Serious & usually fatal
Liver & Renal Tubular Damage
Generalized aminoaciduria
3 types (all produce tyrosylemia & tysosyluria)
13. β
Type 1:
Deficient in Fumarylacetoacetate hydrolase
(FAH)
β Generalized renal tubular disorder & progressive
liver failure
Type 2:
β
β
Deficient in Tyrosine aminotransferase
β Corneal erosion; lesions of palms, fingers, &
soles of feet due to crystallization of tyrosine in
cells
Type 3:
β
β
β
β
Deficient in P-hydroxyphenylpyruvic acid
dioxygenase
Mental retardation if dietary restrions of
phenylalanine & tyrosine not implemented
14. β
Nitroso-naphtol Test
Urinary screening test for tyrosine & its
metabolites
β Nonspecific
β (+) orange-red = indicates need of further tests
1. Place 5 drops of urine in a tube
2. Add 1mL of 2.63N nitric acid
3. Add 1 drop of 21.5% sodium nitrite
4. Add 0.1mL l-nitroso-2-napthol
5.Mix
6. Wait 5 minutes
7. Observe colour
MS/MS screening test
β
β
β
Available for Type 1 & 2
15. Melanuria
β
β
β
2nd pathway of Tyrosine
Responsible for prod. of melanin, thyroxine,
epinephrine, protein & tyrosine sulfate
Major lab concern: Melanin
β
β
β
β
Confers dark hair, eye, and skin colour
Deficieny: Albinism
Elevated: overproliferation of melanocytes =
Malignant Melanoma
Tumor cells secrete 5,6-dihydroxyindole =
oxidizes to melanogen, then melanin =
dark urine
17. Alkaptonuria
β
1 of 6 inborn errors of metabolism
β
3rd major defect in phenylalanine-tyrosine pathway
β
β
β
β
β
Absence of gene needed to prod. Homogentisic acid
oxidase
Homogentisic acid accumulates in blood, tissue, urine
βalkali loverβ - turns dark & alkali after standing in room
temp
Brown pigment deposited in body tissues (esp. ear);
deposit in catilage (arthritis)
Many develop liver & cardiac disorders
18. β
Homogentisic acid reacts w/:
1. Ferric Chloride: transient deep blue colour
2. Clinitest: yellow precipitate
3. Alkali: darkening of urine (interference: large
amounts of ascorbic acid)
Silver nitrate & ammonium hydroxide: black
Read:
Spectrophotometer, Chromatography procedures
19. Branched-Chain Amino Acid
Disorders
β
A methyl group branches from main aliphatic
carbon chain
1. accumulation of 1/more early amino acid
degradation products (maple syrup urine dse)
2. organic acidemias: accumulation of organic
acids further down metabolic pathway
β
Ketonuria in newborn
20. Maple Syrup Disease
β
Rare, Autosomal recessive trait, caused by IEM
β
Involved: leucine, isoleucine, valine
β
β
β
Absence of gene for oxidative carboxylation of keto
acids (alpha-ketoisovaleric, alpha-ketoisocaproic,
alpha-keto-beta-methylvaleric) = accumulation in
blood & urine
Failure to thrive after aprox. 1 week
Maple syrup odor (11th day): due to rapid
accumulation of keto acids
β
REPORT! Prevent severe mental retardation &
death:
21. β
Newborn screening: MS/MS
β
2,4-dinitrophenylhydrazine (DNPH) reaction
β
β
β
β
β
Most frequently performed urinary screening
(+) yellow turbidity or precipitate
Interferece: Large doses of ampicillin
Nonspecific
Treatment can be started based on odor, clinical
symptoms & positive DNPH test while
confirmatory prodecures are being done
24. β
Isovaleric Acidemia:
β
β
Sweaty feet odor of urine
β Caused by accumulation of
isovalerylglycine due to a deficiency of
isovaleryl coenzyme A in the leucine
pathway
Propionic & Methylmalonic acidemia:
β
β
Errors converting isoleucine, valine, threonine,
methionine to succinyl coenzyme A
Immediate precursor to methylmalonic acid:
Propionic acid
26. Purine Disorders
β
Lesch-Nyhan Disease
β
β
β
β
β
β
Sex linked recessive
Massive excretion of uric acid crystals
Absence of Hypoxanthine guanine
phosphoribosyltransferase
Severe motor defects, mental retardation,
tendency towards self-destruction, gout, renal
calculi
First 6-8 months: normal development
First symptom: (uric acid) orange sand in
diapers