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Guanase activity in endemic balkan nephropathy
CLINIC.4
CGA
CHIMICA
445
ACTA
4613
GUANASE
K. PlIODXNO\ Cmtval
Clinical
(Iicceivcd
ACTIVITY
IN
ENDEMIC
BALKAN
NEPHROPATHY
AXUDA. hSTI
Thrrapeutic
Clinic,
Higher
Institute
oflZIediciw,
Sofia
(Bulgavia)
lRIay 17, 1971)
Serum guanase
activity
was studied
in 71 patients
with renal diseases
and in
30 normal individuals. A significant increase of serum guanase activity was recorded in 73.3 per cent of patients with endemic nephropathy. Only in isolated cases of chronic glomerulonephritis and chronic pyelonephritis, guanase activity was shown to be slightly increased. It is admitted that increased guanase activity in patients with endemic nephropathy is due to destructive lesions and cytolysis in the epithelial cells of the renal tubuli and, possibly, in the liver parenchyma as well. No correlation was found between increased serum guanase activity, on the one hand, and the degree of nitrogen retention, glomerular filtration and flocculation tests, on the other.
Guanase is an enzyme of purine metabolism. In man its maximal activity is found in the liver, brain and kidneys; moderate in the small and large intestines; and negligible in all other organs and biological fluids1p5. Guanase appearance in the blood is most probably associated with damage of the first and eventually the second group of organs. Guanase activity has been shown to be a sensitive index of liver cell damag-e3y4,6s7.High enzymatic activity has been found in vira131”T7and toxic hepatitis, in carbon tetrachloride poisoning8-10, after treatment with radioactive goldl’. Guanase activity has almost never been studied in renal disease. This may be explained with the works of Knight et a1.3, who failed to disclose guanase activity in the blood serum of dogs, following experimental intoxication with uranyl-acetate and mercury chloride. They found no increased guanase activity in acute glomerulonephritis as well. Four years later, however, Bastide and Bastidelz observed a constantly increasing enzymatic activity in the liver, kidneys and red cells 24 h after uranylacetate intoxication in rats. These authors failed to establish any increase of the enzyme in the blood at the time of its increase in the renal tissue. In 1969 Kokot et al.‘” studied the plasma guanase activity in patients with acute and chronic renal failure, subjected to hemodialysis or peritoneal dialysis. Decreased enzymatic activity was recorded in part of the patients after hemodialysis. In an extensive study of 500 patients with different diseases French authors failed to detect increased guanase activity in some renal affections without pointing them’. Increased serum guanase activity in
PRODAKOV,
446 renal damage has been established by Al Khalidi are directly responsible for this increase.
et aZ.14 who believe
ASTRUG
that the kidneys
Increased serum guanase activity has been found by Stefanovic et al.‘5 in some kidney patients with destructive lesions-chronicglomerulonephritis and pyelonephritis and other renal diseases, among which two cases of endemic Balkan nephropathy. The present work is the result of a study of guanase activity in some renal diseases. Our attention was focused on endemic Balkan nephropathy since there are almost no reports of systematic studies in this disease. Patients with chronic pyelonephritis and glomerulonephritis served as controls. MATERIAL
AND METHODS
The survey covers 71 patients-42 women and 29 men-from 19 to 79 years of age with the following diseases: endemic nephropathy, 38 patients, chronic pyelonephritis, 23 patients, and chronic glomerulonephritis, IO patients. A control group of 30 healthy blood donors was examined for guanase activity on another occasions. Guanase activity was determined by the spectrophotometric method of Hue and FreeI with some modifications5. RESULTS
The results of serum guanase shown in Table I. TABLE
activity
studies in the two groups of patients
are
I
SERUM GUANASE ACTIVITY Group
IX
SOME
RENAL
Guanase
activity Upto3.ImlJ
Over 3.2 mu
No.
s:,
No.
9,
30
30
IO0
38 23 IO
9 21 9
29
76.3
No.
Controls Endemic nephropathy Chronic pyelonephritis Chronic glomerulonephritis
DISEASES
23.7 91.2
90.0
Mean value
1.5 2
8.8
I
10.0
11.8 0.5 I.1
I.jH
x.73 0.46 I.12
Patients with endemic nephropathy usually exhibit increased serum guanase activity, its mean value being more than IO times as high as that of the other two groups of kidney patients. The differences between these values are statistically significant (P < 0.001). There was no essential difference in serum guanase activity in patients with endemic nephropathy with and without nitrogen retention. Single patients with pyelonephritis or glomerulonephritis showed slightly elevated serum guanase activity. No correlation was established between guanase activity and nitrogen retention, as evaluated by serum creatinine and urea concentrations. Patients with high-grade nitrogen retention showed absence of guanase activity; conversely, in many patients without renal failure guanase activity was increased. We did not find any correlation between enzymatic activity and glomerular filtration, as deterClin. Chim.
Acta,
35 (1971) 445-448
ENDEMIC
BALKAN
NEPHROPATHY
447
mined by endogenous creatinine clearance, neither was any dependence established between flocculation tests and transaminase activity, on the one hand, and guanase activity,
on the other.
DISCUSSIOX
The principal
finding in this study was the increase
of serum guanase
activity
in more than 314 of patients with endemic nephropathy. It is difficult to give an adequate explanation for this phenomenon. Epithelial cell destruction and cytolysis in the renal tubuli, observed in patients with endemic nephropathy, may be held responsible for increased guanase activity. In a histomorphological study of kidney biopsy and necropsy material from patients with endemic nephropathy PetrinskaVenkovska17*1s described severe parenchymatous and lipoid dystrophy, occasionally even with necrobiosis in the epithelial cells of the convoluted tubuli. It seems that under normal conditions guanase, 96y0 of which may be found in the cytoplasm of the kidney cells and only 5% in the nuclei and mitochondrials, with difficulty passes through the cell membrane in the extracellular fluids. This is likely to be due to the size of the enzyme molecule, whose molecular weight has been estimated to approximately 1oo~15o,ooo~~~~~ and to the ability of guanase to form molecular complexes as well. Destructive cellular changes with cytolysis seem to occur in some severe intoxications and affections with toxic genesis, thus enabling guanase emersion in the extracellular fluid. Bell et ~2.7 termed guanase “the cytolysis enzyme”. It may be admitted that increased serum guanase activity in endemic nephropathy is of hepatic origin, since in this disease dystrophic lesions have equally been observed in the liver cellsQ917. Mile+ recorded moderate liver function changes in patients with endemic nephropathy, without disclosing changes in serum transaminase and aldolase activity in these patients. REFERENCES I H. M. KALCKAR, J. Biol. Chem., 167 (1947) 461. 2 R. LEVIZIE, T. HALL AND C. HARRIS, Cancer, 16 (1963) 269. 3 E. KXIGHTS, J. WHITEHOUSE, A. HUE AXD C. SAXTOS, J. Lab. Clin. Med., 65 (1965) 355. 4 E. COODLEY, Amer. J. Gastroenterol., 52 (1969) 189. - K. PRODAXOV, Unpublished data (1969). 2 E. DYACHIN.~, Labor. Delo, 5 (1969) 289. 7 A. BELL, R. DIETSCH, R. ALARY, B. SAYOX, R. LEVRAT, J. NESMOZ AND N. NYSSEN, Presse Me’d., 78 (1970) 495. E. SURMA AND J, KRAWCZYXSKI, Diagn. Labor., I (1966) 29. E. SURMA AXD 1. KRAWCZYNSKI. Clin. Chim. Acta. 16 (1966) 29. K. PROD~OV, ir. KOLARSKI AND Ts. GRANCHAROVA, l&i. ked.. 8 (1969) 233. N. FEODOROV. L. FURAEVA, K. FROMICHENKO AND M. BOGOMAZOV, Labor. Delo, 9 (1969) 339. J. BASTIDE AND P. BASTIDE, Compt. Rend. Sot. Biol., 162 (1969) 1169. FR. KOKOT, J. KUSKA, H. KOZIAK AND M. KOWALIK, Zschr. Iner. Med., 24 (1969) 702. U. A. S. AL-KHALIDI, S. APTIMOS AND N. SANAMUSH-ARRASIEH, Clan. Chim. Acta, 29 (1970) 381. I5 0. STEFANOVIC, M. PUSOVIC, P. TRIIMAC, P. TEPAVCEVIC, D. FELLE AND T. BUNOVIC, Archiv Farm., 20 (1970) 193. 16 A. HUE AND A. FREE, Clin. Chem., II (1965) 708. 17 S. PETRIXSKA-VENKOVSKA, in A. PUCHLEV (Ed), Endemic Nephropathy, M and F., Sofia, 1962, 8 o 10 II 12 13 14
P. 72. 18 S. PETRIXSKA-VENKOVSKA, in A. PUCHLEV (Ed), Morphologic Aspect of Endemic h’ephropathy, I. Inter. Symp. Endem. Nephropathy, Bulg. Acad. Sci. Press, Sofia, 1963, p. 95. 19 K. SUDHIR, 1’. JOSAN, S. SANGER, K. TEWARI AXEDS. KRISHNAN, Biochem. J., 102 (1967) 691. Clin. Chim.
Acta,
35 (1971)
445-448
448
PRODANW’,
ASTRUG
CGA
CHIMICA
445
ACTA
4613
GUANASE
K. PlIODXNO\ Cmtval
Clinical
(Iicceivcd
ACTIVITY
IN
ENDEMIC
BALKAN
NEPHROPATHY
AXUDA. hSTI
Thrrapeutic
Clinic,
Higher
Institute
oflZIediciw,
Sofia
(Bulgavia)
lRIay 17, 1971)
Serum guanase
activity
was studied
in 71 patients
with renal diseases
and in
30 normal individuals. A significant increase of serum guanase activity was recorded in 73.3 per cent of patients with endemic nephropathy. Only in isolated cases of chronic glomerulonephritis and chronic pyelonephritis, guanase activity was shown to be slightly increased. It is admitted that increased guanase activity in patients with endemic nephropathy is due to destructive lesions and cytolysis in the epithelial cells of the renal tubuli and, possibly, in the liver parenchyma as well. No correlation was found between increased serum guanase activity, on the one hand, and the degree of nitrogen retention, glomerular filtration and flocculation tests, on the other.
Guanase is an enzyme of purine metabolism. In man its maximal activity is found in the liver, brain and kidneys; moderate in the small and large intestines; and negligible in all other organs and biological fluids1p5. Guanase appearance in the blood is most probably associated with damage of the first and eventually the second group of organs. Guanase activity has been shown to be a sensitive index of liver cell damag-e3y4,6s7.High enzymatic activity has been found in vira131”T7and toxic hepatitis, in carbon tetrachloride poisoning8-10, after treatment with radioactive goldl’. Guanase activity has almost never been studied in renal disease. This may be explained with the works of Knight et a1.3, who failed to disclose guanase activity in the blood serum of dogs, following experimental intoxication with uranyl-acetate and mercury chloride. They found no increased guanase activity in acute glomerulonephritis as well. Four years later, however, Bastide and Bastidelz observed a constantly increasing enzymatic activity in the liver, kidneys and red cells 24 h after uranylacetate intoxication in rats. These authors failed to establish any increase of the enzyme in the blood at the time of its increase in the renal tissue. In 1969 Kokot et al.‘” studied the plasma guanase activity in patients with acute and chronic renal failure, subjected to hemodialysis or peritoneal dialysis. Decreased enzymatic activity was recorded in part of the patients after hemodialysis. In an extensive study of 500 patients with different diseases French authors failed to detect increased guanase activity in some renal affections without pointing them’. Increased serum guanase activity in
PRODAKOV,
446 renal damage has been established by Al Khalidi are directly responsible for this increase.
et aZ.14 who believe
ASTRUG
that the kidneys
Increased serum guanase activity has been found by Stefanovic et al.‘5 in some kidney patients with destructive lesions-chronicglomerulonephritis and pyelonephritis and other renal diseases, among which two cases of endemic Balkan nephropathy. The present work is the result of a study of guanase activity in some renal diseases. Our attention was focused on endemic Balkan nephropathy since there are almost no reports of systematic studies in this disease. Patients with chronic pyelonephritis and glomerulonephritis served as controls. MATERIAL
AND METHODS
The survey covers 71 patients-42 women and 29 men-from 19 to 79 years of age with the following diseases: endemic nephropathy, 38 patients, chronic pyelonephritis, 23 patients, and chronic glomerulonephritis, IO patients. A control group of 30 healthy blood donors was examined for guanase activity on another occasions. Guanase activity was determined by the spectrophotometric method of Hue and FreeI with some modifications5. RESULTS
The results of serum guanase shown in Table I. TABLE
activity
studies in the two groups of patients
are
I
SERUM GUANASE ACTIVITY Group
IX
SOME
RENAL
Guanase
activity Upto3.ImlJ
Over 3.2 mu
No.
s:,
No.
9,
30
30
IO0
38 23 IO
9 21 9
29
76.3
No.
Controls Endemic nephropathy Chronic pyelonephritis Chronic glomerulonephritis
DISEASES
23.7 91.2
90.0
Mean value
1.5 2
8.8
I
10.0
11.8 0.5 I.1
I.jH
x.73 0.46 I.12
Patients with endemic nephropathy usually exhibit increased serum guanase activity, its mean value being more than IO times as high as that of the other two groups of kidney patients. The differences between these values are statistically significant (P < 0.001). There was no essential difference in serum guanase activity in patients with endemic nephropathy with and without nitrogen retention. Single patients with pyelonephritis or glomerulonephritis showed slightly elevated serum guanase activity. No correlation was established between guanase activity and nitrogen retention, as evaluated by serum creatinine and urea concentrations. Patients with high-grade nitrogen retention showed absence of guanase activity; conversely, in many patients without renal failure guanase activity was increased. We did not find any correlation between enzymatic activity and glomerular filtration, as deterClin. Chim.
Acta,
35 (1971) 445-448
ENDEMIC
BALKAN
NEPHROPATHY
447
mined by endogenous creatinine clearance, neither was any dependence established between flocculation tests and transaminase activity, on the one hand, and guanase activity,
on the other.
DISCUSSIOX
The principal
finding in this study was the increase
of serum guanase
activity
in more than 314 of patients with endemic nephropathy. It is difficult to give an adequate explanation for this phenomenon. Epithelial cell destruction and cytolysis in the renal tubuli, observed in patients with endemic nephropathy, may be held responsible for increased guanase activity. In a histomorphological study of kidney biopsy and necropsy material from patients with endemic nephropathy PetrinskaVenkovska17*1s described severe parenchymatous and lipoid dystrophy, occasionally even with necrobiosis in the epithelial cells of the convoluted tubuli. It seems that under normal conditions guanase, 96y0 of which may be found in the cytoplasm of the kidney cells and only 5% in the nuclei and mitochondrials, with difficulty passes through the cell membrane in the extracellular fluids. This is likely to be due to the size of the enzyme molecule, whose molecular weight has been estimated to approximately 1oo~15o,ooo~~~~~ and to the ability of guanase to form molecular complexes as well. Destructive cellular changes with cytolysis seem to occur in some severe intoxications and affections with toxic genesis, thus enabling guanase emersion in the extracellular fluid. Bell et ~2.7 termed guanase “the cytolysis enzyme”. It may be admitted that increased serum guanase activity in endemic nephropathy is of hepatic origin, since in this disease dystrophic lesions have equally been observed in the liver cellsQ917. Mile+ recorded moderate liver function changes in patients with endemic nephropathy, without disclosing changes in serum transaminase and aldolase activity in these patients. REFERENCES I H. M. KALCKAR, J. Biol. Chem., 167 (1947) 461. 2 R. LEVIZIE, T. HALL AND C. HARRIS, Cancer, 16 (1963) 269. 3 E. KXIGHTS, J. WHITEHOUSE, A. HUE AXD C. SAXTOS, J. Lab. Clin. Med., 65 (1965) 355. 4 E. COODLEY, Amer. J. Gastroenterol., 52 (1969) 189. - K. PRODAXOV, Unpublished data (1969). 2 E. DYACHIN.~, Labor. Delo, 5 (1969) 289. 7 A. BELL, R. DIETSCH, R. ALARY, B. SAYOX, R. LEVRAT, J. NESMOZ AND N. NYSSEN, Presse Me’d., 78 (1970) 495. E. SURMA AND J, KRAWCZYXSKI, Diagn. Labor., I (1966) 29. E. SURMA AXD 1. KRAWCZYNSKI. Clin. Chim. Acta. 16 (1966) 29. K. PROD~OV, ir. KOLARSKI AND Ts. GRANCHAROVA, l&i. ked.. 8 (1969) 233. N. FEODOROV. L. FURAEVA, K. FROMICHENKO AND M. BOGOMAZOV, Labor. Delo, 9 (1969) 339. J. BASTIDE AND P. BASTIDE, Compt. Rend. Sot. Biol., 162 (1969) 1169. FR. KOKOT, J. KUSKA, H. KOZIAK AND M. KOWALIK, Zschr. Iner. Med., 24 (1969) 702. U. A. S. AL-KHALIDI, S. APTIMOS AND N. SANAMUSH-ARRASIEH, Clan. Chim. Acta, 29 (1970) 381. I5 0. STEFANOVIC, M. PUSOVIC, P. TRIIMAC, P. TEPAVCEVIC, D. FELLE AND T. BUNOVIC, Archiv Farm., 20 (1970) 193. 16 A. HUE AND A. FREE, Clin. Chem., II (1965) 708. 17 S. PETRIXSKA-VENKOVSKA, in A. PUCHLEV (Ed), Endemic Nephropathy, M and F., Sofia, 1962, 8 o 10 II 12 13 14
P. 72. 18 S. PETRIXSKA-VENKOVSKA, in A. PUCHLEV (Ed), Morphologic Aspect of Endemic h’ephropathy, I. Inter. Symp. Endem. Nephropathy, Bulg. Acad. Sci. Press, Sofia, 1963, p. 95. 19 K. SUDHIR, 1’. JOSAN, S. SANGER, K. TEWARI AXEDS. KRISHNAN, Biochem. J., 102 (1967) 691. Clin. Chim.
Acta,
35 (1971)
445-448
448
PRODANW’,
ASTRUG