Beta-glucuronidase studies in women

IV. The Significrtnce S. C.

of the Leucocytie Popda.ticm

M.D.,

KASDON, W.

El.

E.

ROMSEY,

FISHMAN,

A.B.,

PH.D.,

in Veal

I?. HOMBUR~ER, BOSTON,

M.D.,

AND

MASP.

(Prom the Cancer Research, and Ca,noer Control Unit, Departme&s of Swrgeq, lkchemistry, Tqfts Co!Tege Nedical ScS,hool I

A

BltM*

A4edicAv,e,ccnd

TTENTION

has been cent,ered on the role and sourcc~of the enzyme hetaglucuronidase in physiological and pathological st-ates. 1-n earlier publications,‘, n control values for vaginal fluid and uterine cervix tissues in health and disease have been presented. It has been demonstrated that the blind vaginal pouch, with the uterus and cervix ablated, is frequently rich in betaglucuronidase activity. Two obvious possible sources of the enzyme in vaginal fluid, Trichomonas ~raginalis and the presence of blood elements, were considered. An evaluation has been made of the significance. of the Trichomonas growing in the vaginal fluid as a contributor* lo the euzymr conlc~nt of vagina,I fluid+” The present study investigates the Ir~lcocytes and their relation to vaginal fluid beta-glucuronidase activity. Sinctl previous data on human blood and lymph4 showed low enzyme activity in these substances, not esceeding 30 to 30 units per gram, the leucocytes,*. ’ averaging nrountl 1.OOOunits per gram, remain the most probable extraneous SOUIW rich iu Ijct~~-glucuronidasc. Plan of Study Vaginal fluid has been collected in the manner previously described’ h> means of the standard Papanicolaou pipette and bulb from lOO-nonpregnant, women without cancer of the genital tract. Standard smears were made as fol routine exfoliative cytological study from an aliquot of the specimen used for enzyme determinations. The smears were then stained according to Papanicolaou’s trichrome technique.G The number of leucocytes for one hundred epithelial cells from representative fields on each slide were counted in order to arrive at a reasonable estimate of the density of leucocytes in each specimen. Multiple counts on slides with large leucocyte populations showed a variation of less than 15 per cent in the total number of leucocytes per 100 epithelial cells. The number of leucocyt.es per 100 epit,helial cells was then plotted against the beta-glucuronidase activity of the identical specimens in order to establish the relationship between the two. *This study was aided by grants from the United American Cancer Society, ‘Inc., New York, an& tt-le Damon Research, New York.

II I:!

States Runyon

Public Health Service, the Memorial P%md for Cancer

BETA-QLUCURONIDASE

STUDIES

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1143

WOMEN

Results and Comment It can be seen from Fig. 1 that there is drift upward when the average number of leucocytes per 100 epithelial cells is plotted against the betaglucuronidase activity in ranges of from 50 units per gram up to 400 units per gram. The scattergram of values upon which the curve was based is superimposed upon Fig. 1. Above the last-named level, 17 observations were made in which the vaginal fluid beta-glucuronidase activity ranged regularly from 500 to 1,500 units and here the average leucocyte count per 100 epithelial cells was 394. AVERAGE PER

ICO

LEUCOCYTE EPITHELIAL

COUNT CELLS

17 OBSERVATIONS FROM

300

500

WITH ACTIVITIES

T O 1500

UNITS

AVERAGE 394 : 100 EPITHELIAL

LEUCCCYTE CELLS

400

500

PER

GRAM

COURT PER

:::: I 100

P-GLUCURONIDASE

AVERAGE

200

300

UNITS

ACTIVITY

PER GRAM

VALUES Fig.

1.

Although in the beta-glucuronidase range of 0 to 300 units per gram of vaginal fluid there is a correlation with the leucocyte population, the great number of specimens in which large numbers of leucocytes were not paralleled by high enzyme titers (SC. scattergram) makes it seem unlikely that the leucocyte population is an important source of vaginal fluid glucuronidase. Moreover, the vaginal fluids rich in beta-glucuronidase did not contain leucocytes in proportion to their activity (Table I). The fact that so many vaginal fluids containing large numbers of leucocytes did not exhibit more than.300 units per gram of activity makes it further probable that the leucocyte population is of minor importance as a source of beta-glucuronidase in vaginal fluid. Summary

and Conclusions

Vaginal fluid specimens were collected from 100 women and were studied with respect to leucocyte population and vaginal beta-glucuronidase activity. Counts of leucocytes per 100 epithelial cells were made and related to the enzyme activity of the vaginal fluid. It was found that as the beta-glu-

Am. 1. Obst. & Gym: May. 195

Volume 61 Number 5

BETA-GLUCURONIDASE

STUDIES

IN

WOMEN

1145

curonidase activity rose from 0 to 300 units per gram, the leucocyte count similarly increased. However, this relationship was not linear but leveled off in the range of activity from 300 to 1,500 units per gram. There were many vaginal fluids which were rich in leucocytes and poor in beta-glucuronidase activity and conversely there were instances where a high beta-glucuronidase activity was associated with a low leucocyte count. It is concluded that the leucocyte population is of some minor importance as a source of beta-glucuronidase in the vaginal fluid. References 3. Fishmao, 2. Kasdon, 3. Fishman,

W. H., Kasdon, S. C., Fishman, W. H., Kasdon,

J. OBST. 4. Fishman, W. 5. Anlyan, A. J., 6. Papani&laou,

& GYNEC.

5. C., and Homburger, F.: J. A. M. A. 143: 350, 1950. W. H., and Homburger, F.: J. A. M. A. 144: 892, 1950. S. C., Bonner, C. D., Fishman, L. TV., and Homburger, F.: 6;1:

647,

1951.

H., Springer, B., and Brunetti, Gamble, J., and Hoster, H. A.: G. N.: Science 95: 438, 1942.

R.: J. Biol. Chem. 173: Cancer 3: 116, 1950.

449,

1948.

AM.