Serum deoxyribonuclease I and II in pathologic conditions other than pancreas diseases

\‘()I.. 4

(Iqy))

SERUM I)NaSe

KiL 4 (‘959)

1 AND ff

IN PATHOLOGIC CONDITIOX3

193

acid (TCA) 3.6 M. After shaking and centrifuging, 0.3 ml of the clear supernatant fluid is withdrawn; deoxyribose is assayed according to the classic DISCHE method 1%. For the assay of DNase II acetic buffer (PH 5.6) is used instead of veronal, and the D&S. is dissolved in twice-distilled water only. The details of the procedure are reported in Table I: TABLE ”..-..A .~. f r

.l.ll-_.~-.._ Serum (ml) Buffer PH 7.4 (m’) Buffer 1’~

5.6

RNA l&U

I

1

DNase II -..._ A c R -. “-_-. _____.._“.___.~ I I 1

_

I

I

I

0.5 -

a.5

o-5

_

x 0.8

I

DNase --3

c

..__

1 _

(ml)

(ml) (ml)

o”5 -

0-S -

0.5

prrt in a water bath at 37O for 4 i; I

Serum (ml) TCA (ml)

,s

a.8

0.8

0.8

0.8

shake, centrifuge and add to 0.5 ml of clear supernatant Dische (ml)

I

I

1

I

1

fluid I

put in a boiling water bath for IO min and then add x.5 ml of sulfuric acid x.50& in acetic acid. ll-_--._----

____

---

~_

..--I___

. ..

On a spectrophotometer (at 625 rnp wavelength) A and B are examined with C as blank and the extinction values are recorded. The number of ,ug of deoxyribose released by the enzymic reaction is obtained from the difference between A and 3 ; this correction is necessary in view of the fact that commercial preparations of deoxyribonucleic acid contain variable amounts of deFolymerized products. TABLE SERUM

-.

..-

.%&jects

_--..

ACTIVITY -

-“--.

No. Mean

_ _. ._______ __I_-_.^ l__l---21 Normal contraIs Malignant disease 46 Liver disease 37 Misc. pathologic conditions 13

II

DNsse I

20.8

7.6 19.3 8.0

_~..._.__ O--IO x0-20 -_____ SE0 “/A 920 y; _i_~--__-________-I6 29 9 43 o-59.4 o-59.4 36 78 6 ‘3 18498224111 o-99.0 9 69 2 15 o-36.3 Range

Deoxyribose (pg) .-. -I-_..__-_ _.._ -.-_-.i._.__ 20-30 30-40 40-50 .p-& 60-70 >70 -_._.._- _ - _ A0 “i; %O y& -3E*pa n” $4 iz@ 7; we “‘0 I I

5 2

_

I

TABLE .___---.

_______-,.--.

_______^l-.-. Subjects

l.“_-

No. Mean

Range

.--._-l

Normd cm~tiols Malignant disease Liver disease Misc. pathologic conditions ..-.-. ______ References p. r96

20 46 37 13

18.2 9.3 x2.9 4,o --

*39.6 o-67.2 o-125.4 o-23.2 ____~

_

5 L _

-.__...

2 L -

IO - 4 -1338 _.

I

-

-_

.-_--._

-

-

_ -..

_“_

III

SEHUFFf DNase II I ...l..-_~

Ij

_~_l--

.~._______I_

.

2 TO I -‘I r 3252

ACTIVITY ___-

_“___..___

. ..^. .I

., __

._~

..__

D~o~~ribase (pg,l

O--IO 10-20 20-30 30-40 40-50 50-60 60-70 >7o ~-----______no % VP 24 no % n0 :& no yO no y. vzO 56 no y. -l_l --. 8 40 8 40 I 5 3 x5 - - -- - - 29 63 12 26 4 9 -- - - -.. f 2 23 62 IO 27 I 3 I 3 - - -- -- 2 5 g 1 8 - -. - II 85 I ._.. I..._.-..~__.. ” . ..__._ . ..-_ _. __

The calibration chart may be obtained by using pure dtoxyribose. Ikzymic activity is expressed as ,~g of dcoxyribose released from I ml of serum. By using progressive dilution of pure l)Kasc I (Mann) we found that 100 pg of dcosyribosc~ arc‘ released by a solution containing 2.5 ,LL.~ of Dru’ase/ml. I)Nasc I and II have been measured in the scr-a of 117subjects; 21 M’C’ICnormal controls, students and physicians, 46 were patients with malignant proliftrative disease (27 untreated carcinomas, 8 carcinomas after SIC treatment, II lcukcmias and Hodgkin’s discsase), 37 were patients with liver disease (12 cirrhosis, 10 obstructive. jaundice and 6 viral hepatitis) ulwr, congcstivc hart failuw,

The results

arc rcportcd

; 13 additional

patients infarction).

myocardial

had various diseases

(gastric

in Tables II and I II and in Figs. I and 2. The following

conclusions can be drawn : I) \Vith simultaneous determinationsno correlation and DIGase II activity, in either normal or pathological 2) There is a wide range of values in all groups.

can befoundbetwetn conditions.

I)h’asc 1

3) In a large number of patients with malignant diseases low values both of I)Nasc I and I)Sasr II have been observed. M’hereas only z8.6”,, (I)i\;ase I) and L+o”(,

A

B

C

0

E

F

100

m

90 60

E

70

$ 0

$

60

g

50

!$

40

2

30

z: s

20 10 0

G

H

. . . .

..

..

. . . . .. .. .. .. . * ,...

. . I.....

.. . . .*. : . . . . .... . ...”

.

.

. . .

..

l

m.....

-

(DBase II) of normal controls show values between o and IO pg, up I) and 63’:h (I>?rTase II) of the cases with cancer are found in this low As to the DNase I the difference has been proved to be statistically 15.387; P 0.001). Ko definite conclusions can be drawn in regard observed x2 value having a P between 0.10 and 0.05. 4) Only occasionally very high values have been found (above of 60 ,q of deoxyribose for DiK’ase I and 40 ,~g for DNase II). These in patients with a very severe degree of hepatic damage. Rt+ences p. ryfi

to 78.2’J,J (Dru’ase range of activity. significant (x2 to DNasr II, the

the normal limits were consistently

VOL. 4 (1959)

SERUM

DNase I AND A

x 0 _m 0: z& : !2 ; I: d la

-

130 120 110 100 90 80 70 60 50 40 . 30

6

-

II IN PATHOLOGICCONDITIONS -

C

-

0

E

-

F

G

-

-

195

H

.

.

. l

*

.

.

.

20 I :.;. __

.

. .. l

.. . . -

0

’ . ,

. I. ... .

. . l

*

I...

.

a**

. . . .

. . a.:.

.

Fig. a. DNase II activity (A = normal controls; B = carcinomas; C = carcinomas after xR treatment: D = leukemias and Hodgkin’s diseases; E = cirrhosis; F = obstructive jaundice; G = hepatitis: H = various pathologic conditions). COMMENT A significative

increase

of pancreatic

DNase

activity

has recently

been shown

in acute pancreant as well as in pancreatic necrosis experimentally induced with ethioninel or COXSACKIE virus 17. The present data show that increased DNase I and DNase II activities cannot be considered characteristic of any other disease. Occasional high values of both enzymes have been observed in a few cases of advanced liver insufficiency with severe jaundice. According to KURNICK AND CARRERAis the liver might withdraw the enzyme from the blood stream and thus regulate its plasma level: a decreased clearing function would thus follow liver impairment. They observed also that serum DNase (DNase I) is considerably elevated in parenchymatous liver disease but not in obstructive icterus in man. The present data do not substantiate these results. Therefore the determination of serum levels of these enzymes would not seem to be as reliable a guide for the evaluation of liver damage as the use of other enzymes (phosphatases, transaminase, aldolase, phosphoisomerase, phosphoglyceromutase) . Significantly low values of DNase I have been observed by WROBLESKI AND BODANSKY~~ in 50 patients with malignant diseases as compared with normal individuals of the same age; no significant changes have been observed by KURNICK”. The present results show a statistically significant decrease of DBase I. Since many of these patients have very low values, further investigation is needed to ascertain the possible relationship of level with size and type of tumor. SCHREIDER et aLlo have reported increased values of both enzymes in children irradiated for extensive tumors. Our limited data on irradiated adult subjects seem to conflict with the previous report but need further study. ACKNOWLEDGEMENT The authors for his assistance References p.

196

wish to thank and criticism.

Prof. O.C. Dogliotti,

Director

of the Medical Clinic,

sl.nlnl.9K\ Serum dcoxyril~onucleast I and II have heen studied in a group of patients with liver disease, carcinomas and miscellaneous conditions. Since both enzymes have lwcu found in human w-a, their beha\-ior in pathological conditions may 1~ of interest and c~vcntually of diagnostic value. .j wide range of valuc~ has lwcn found in all groups with

no correlation

lwtn

found

sionally

lwtwcen

in patients

okrved

with

I)Saw

I and

maliqxmt

in casc5 with

advanced

l)Xaw

diseases liver

I I. Significantlv

whereas

high

insut’hcicticy.

values

low ~xlues have

liavc~

lwcn occa-