1 proteinuria. analysis of urine seminar no. 11 - chapter 12 -
TRANSCRIPT
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Proteinuria. Analysis of urine
Seminar No. 11
- Chapter 12 -
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Filtration of plasma
• glomerular membrane is a filter system
• structural barrier (collagen IV)
• electrostatic barrier (neg. charges of sialic acid in
glycoproteins repulse anionic macromolecules - proteins)
• the basement membrane allows free movement of
electrolytes, water and small molecules (urea, glucose, AA,
creatinine ...)
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The filtration of proteins strongly depends
on their molecular mass
• Proteins with Mr < 60 000 (microproteins)
pass easily into urine regardless of their charge
• Proteins with Mr 60 000 – 150 000
only very small amount is filtered into urine
• Proteins with Mr > 150 000
do not pass into urine regardless of their charge
scheme on p. 69
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Q.
How are proteins resorbed from primary urine?
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Resorption of proteins
• microproteins (Mr < 60 000) are resorbed from the primary
urine by pinocytosis
• after hydrolysis in tubular cells are returned into AA pool
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Q.
Which enzyme is readily filtered into urine?
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A.
• see p. 26
• amylase (Mr ~ 50 000)
• catalyses the hydrolysis of starch into dextrins and maltose
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Q.
What is normal daily excretion of
a) proteins b) albumin?
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A.
Daily excretion of proteins
total proteins: < 150 mg/day
albumin: < 20 mg/day
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Q.
What are chemical tests for proteins in urine?
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1. Test strips based on protein error of acid-base indicators
primarily
specific for albumin (albuPhan,
Pliva-Lachema, Brno)
2. Precipitation tests
traditional test performed in a test tube
proteins are precipitated (denaturated) by 5-
sulfosalicylic acid as fine particles (turbidity)COOH
OH
HO3S
see lab manual
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Q.
What is microalbuminuria?
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A.
• urine excretion of albumin in the range
of 20 – 300 mg/day
• an early indicator of diabetic nephropathy
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Q.
Explain how microalbuminuria can be detected?
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A.
• immunochemical tests using antibody against human
albumin
• antibody can be gold-labelled red coloured zone
on a strip corresponds with albumin concentration in
urine sample (test strips Micral)
• antibody freely soluble nephelometry (p. 10)
see lab manual
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Q.
What is SDS-PAGE?
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A.
• sodium dodecyl sulfate polyacrylamide gel electrophoresis
• for principle see p. 7
• a modification of simple electrophoresis
• separation according to molecular mass to distinguish
individual proteins see scheme on p. 70
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Glomerular proteinuria
• consequence of the loss of glomerular membrane integrity
• typical protein: albumin
• selective glomerular proteinuria:
proteins with Mr 60 000 – 100 000 pass into urine
• non selective glomerular proteinuria:
more severe glomerular lessions
proteins of all sizes Mr > 60 000 pass into urine
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Tubular proteinuria
• tubules are unable to reabsorb proteins
• small proteins molecules (microproteins) Mr < 60 000
appear in the urine
• typical protein: β2-microglobulin
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Urine sediment
• suspension material obtained by centrifugation of fresh
urine sample under defined condition
• semiquantitative quantitative
• chemical sediment – crystals of various compounds (salts)
• biological sediment – cells (RBC,WBC), casts, bacteria
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Factors involved in renal stones formation
• increased concentrations of certain ions (Ca2+, Mg2+ ...)
• extreme values of pH of urine
• low intake of fluid (low diuresis)
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Insoluble compounds - Complete
CompoundInsolubility pH range
Acid Neutral Alkaline
Uric acid
Ammonium hydrogen urate
Calcium oxalate
Calcium hydrogen phosphate
Hydroxyapatite
Ammonium magnesium phosphate
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Insoluble compounds
CompoundInsolubility pH range
Acid Neutral Alkaline
Uric acid
Ammonium hydrogen urate
Calcium oxalate
Calcium hydrogen phosphate
Hydroxyapatite
Ammonium magnesium phosphate
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Q.
What is uric acid?
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Uric acid in lactim form is a diprotic acid
N
N
N
NH
OH
HO
OHN
N
N
NH
OH
HO
ON
N
N
NH
OH
O
O
uric acid (lactim) hydrogenurate urate
pKA1 = 5.4 pKA2 = 10.3
2,6,8-trihydroxypurine
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Q.
Give the formulas of compounds:
Calcium oxalate
Calcium hydrogen phosphate
Hydroxyapatite
Ammonium magnesium phosphate
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A.
Calcium oxalate Ca(COO)2
Calcium hydrogen phosphate CaHPO4
Hydroxyapatite Ca5(PO4)3OH
Ammonium magnesium phosphate NH4MgPO4
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Some food components conduce to urolithiasis
Food Commentary
Meat (excess)
Milk products
Mineral waters
Spinach
Rhubarb
Vitamin C (excess)
cystein catabolized to SO42- + 2H+ urine acidification
rich in Ca2+ and phosphates
some of them are rich in Ca2+ and Mg2+
contains oxalic acid (up to 1 %)
contains oxalic acid (up to 1 %)
catabolized to oxalic acid
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Rhubarb (Rheum rhaponticum)
• a herb with long reddish leaf-stalks, rich in oxalic acid
• rhubarb juicy stalks are edible when cooked and sweetened
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Q.
Why is urine alkaline in urinary infections?
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Bacterial urease catalyzes the hydrolysis of urea
H2N-CO-NH2 + H2O 2 NH3 + CO2
NH3 + H2O NH4+ + OH-
alkaline urine concrements of CaHPO4
ammonia
ammonium
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Q.
Which types of stones occur most frequently
in European population?
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A.
• oxalates
• see the table in lab manual, chapter 11
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Be sure to have the index
on the next Monday
May 29, 2006
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Some students have absences
Student Missed seminars
Botonaki F.
Cheema S.S.
Maloiso C.U.
Nikolaides E.
Sulaibeekh S.
Tandail M.
Utulu D.I.
No. 9
No. 4 + TEST, No. 5
No. 9
No. 9
No. 4 + TEST, No. 5
No. 4 + TEST
No. 5
they are strongly advised:
• to make them up in the written form (= answer questions from the corresponding chapter in seminar book)
• to write the 1st revision test, preferably on next Monday, after regular seminar (Cheema, Sulaibeekh, Tandail)