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Alpha -Antitrypsin in Cervical Mucus
Alpha -Antitrypsin in Cervical Mucus GEBHARD F. B. SCHUMACHER, M.D., F.A.C.O.G.,* and MANUEL J. PEARL, M.D., F.A.C.O.G.
of human serum allows rough differentiation of albumin, a1- and a2-globulins, fi-globulins, and y-globulins according to their electrophoretic mobility. These fractions, except albumin, are nonhomogeneous. They consist of several subfractions with individual molecular characteristics. High-resolution systems and immunochemical methods permit further differentiation. More than 30 proteins, glycoproteins, and lipoproteins are now well characterized. The major component of a group of glycoproteins and lipoproteins that migrate with the electrophoretic mobility of a1-globulins is a1-antitrypsin. Its synonyms are al-3,5-glycoprotein, au-globulin, and a,-seromucolde de Schultze. 1, 5, 6,13,14 This a1-globulin is a glycoprotein (12% carbohydrate) with a molecular weight of 45,000, which is small as compared with those of other serum proteins. It inhibits trypsin and chymotrypsin and has therefore been named al-antitrypsin. 13 , 14 Its capacity to inhibit trypsin is in the range of 0.25-0.50 mg. trypsin per milligram of a1-antitrypsin. ac antitrypsin also inhibits autoactivated plasmin but not streptokinaseactivated plasmin. 15 The normal range of this protein appears to be between 2 and 5 mg.jml. of serum. 2 ,13,15 The immunoelectrophoretic analysis of cervical mucus indicates in most normal specimens the presence of acantitrypsin (Fig. 1), as previously reported. 18 If developed with antiserum against total human serum, the immunoelectrophoretic pattern shows five precipitin lines. Specific antiserum against purified a1-antitrypsin identifies the precipitin line in the a1-globulin area as a1-antitrypsin.17,18 The other bands indicate the presence of prealbumin, albumin, transferrin, and 7S y-globulin, as previously ELECTROPHORETIC SEPARATION
From the Division of Laboratories and Research, New York State Department of Health, and from the Departments of Obstetrics and Gynecology, and of Biochemistry, Albany Medical College, Albany, N. Y. Presented at the 23rd Annual Meeting of The American Fertility Society, Washington, D. C., Apr. 14--16, 1967. We thank Miss Margarita Feifel for her technical assistance. Supported in part by Grant IROI HD 02682-01 from the U. S. Public Health Service. *Present address: University of Chicago, Department of Obstetrics and Gynecology, The Chicago Lying-in Hospital, Chicago, Ill.
91
92
SCHUMACHER
&
PEARL
FERTILITY
& STERILITY
Fig. 1. Immunoelectrophoresis of cervical mucus.
reported by Moghissi and Neuhaus l l and by Heinen. The trypsin-inhibitor activity of !Xl-antitrypsin in cervical mucus has been established16 by a combined immunoelectrophoretic and enzyme diffusion technic in fibrincontaining agar gel,3 Immunoelectrophoretic technics of this type provide only qualitative information. The quantitative assessments of proteins present in cervical mucus would be highly desirable to gain insight in secretory functions under the influence of hormones and drugs and under pathologic conditions. METHODS
Immunochemical micromethods are the methods of choice to determine concentrations of soluble proteins in cervical mucus specimensY' 19-21 A simple and accurate method of high sensitivity is the single-radial-immunodiffusion technic in agar gel, introduced a few years ago by Mancini et ap· 8 The principles of this method are illustrated in Fig. 2. A commercially available antiserum* against !Xl-antitrypsin is incorporated into a buffered, uniformly thin, agar gellayer.t The cervical mucus is deposited *Hoechst Pharmaceutical Co., Kansas City, Mo. tDetails of the preparation of the gel are being published elsewhere." Ready-to-use "Partigen"-Immunodiffusion Plates are now also available from Hoechst Pharmaceutical Co.
VOL.
19, No.1, 1968
aI-ANTITRYPSIN
93
into wells, which are punched in the gel with stainless-steel well cutters of 3-mm. diameter.* The lXI-antitrypsin diffuses (with other soluble compounds) radially into the antibody-containing agar, forming a growing disc of antigen-antibody precipitates. The growth of the discs is a function
ANTIGEN SOLUTION
Fig. 2. Principles of singleradial-immunodiffusion technic in agar gel.
ANTIBODY' CONTAINING AGAR
_lADIA~~~~;'ON 8J.dFJ m IDE
O-TIME
AFTER MINUTES
AFTER HOURS
of antigen concentration, of time, of antigen-antibody ratio, and of the diffusion coefficient of the diffusing molecules. The diameter of the discs can be measured and related to standard solutions of purified IXl-antitrypsint Figure 3 shows the results of an experiment with standard solutions of known concentrations. The square of the radius of the discs plotted against the concentration of the standard solutions shows almost a straight line in concentrations between 10 and 1000 /Lg. jml. if the precipitates are measured after 72 hr. of diffusion at room temperature. The r2 values of the discs produced by cervical mucus specimens can be evaluated now by this standard curve. Each series of lXI-antitrypsin determinations requires the use of these standard solutions. Specimens and standard solutions in small portions are stored at 20° C. RESULTS
Cervical mucus specimens from normal healthy women during ovulatory cycles and under the influence of hormonal contraceptives have been investigated by means of this immunologic method for determining the IX]antitrypsin content. A typical picture of an ovulatory cycle is shown in Fig. 4. The lXI-antitrypsin concentration decreases from high postmenstrual values to very low values between Days 11 and 13 and increases again remarkably on Day 14. The low values coincide with high spinnbarkeit values, positive Fern test, very low cellularity, mucorrhea, and positive Sims-Huhner tests. According to these parameters and to the increase of *Immunodiffusion Set, LKB-Instruments, Inc., Rockville, Md. tThe purified ",,-antitrypsin was made available by the courtesy of Dr. Schwick, Behringwerke, Marburg, Germany.
94
SCHUMACHER
&
PEARL
FERTILITY
&
STERILITY
basal body temperature, one can assume that ovulation has occurred on about the twelfth day. Figure 5 shows the preovulatory and postovulatory courses of aI-antitrypsin, spinnbarkeit, and basal body temperature (BBT) of 10 patients. In three of these patients, ovulation was induced by administration of gonadotropins. The day of minimum acantitrypsin values, which ranges between Days 10 and 16 of the spontaneous cycles, has been defined in this
2 YtHrs.
,50 100
,500
Fig. 3. Results of experiment using standard solutions of known concentrations in single-radial-immunodiffusion technic. The growth of the precipitin discs over a period of 72 hr. of diffusion is demonstrated.
VOL.
19, No.1, 1968
95
lq -ANTITRYPSIN
graph as Day o. The days prior to the day of minimal aI-antitrypsin values and the following days are arranged in order as minus or plus days. The spinnbarkeit and BBT values are arranged corresponding to their respective aI-antitrypsin values. The BBT curves and other clinical data give evidence for biphasic cycles in all cases and suggest that ovulation did occur. Two of these individuals became pregnant during the cycle studied. Minimum values of aI-antitrypsin concentration and maximum values of spinnbarkeit
5
o
GOO 500 400
300
200 100
Menses
o
3 4 5 6
~~
9 10 1{12 13 14 IfilS 17 18 19 20 2122
000 00,0
..
a 2425 262728C~
···Qh; .·cdaJj
···fMMUNOPREClPITATES oFD(;"ANTI TRYPSiN;>: Fig. 4. The aI-antitrypsin content of cervical mucus, fem test, presence of cellularity, occurrence of mucorrhea, Sims-Huhner test results, spinnbarkeit, and BBT during ovulatory menstrual cycle.
96
SCHUMACHER
& .PEARL
F'ERTILITY
& STERILl'ry
are quite well correlated and coincide with the last day prior to the stepwise increase of basal body temperature. Therefore, one may assume that ovulation occurred on Day 0 or on Day +1 in some cases. If one compares these three parameters, it appears that the al-antitrypsin values show only "F 99
Basal bady temperature 98 975 97
C:Oj
Spinnbarkeit
10
5
o
___
l-~../
I'o/ml
700
600 500 400 300
200
0(,
-Antitrypsin
100
Fig. 5. Pre- and postovulatory courses of al-antitrypsin, spinnbarkeit, and BBT in 10 patients. G indicates cycles in which ovulation was induced by administration of gonadotropins. Arrow indicates day of minimum al-antitrypsin concentration.
little increase during the first 48 hr. after the minimum on Day 0, while the changes in the spinnbarkeit values and in the BBT values are more pronounced. From the second or third dayan, significantly higher al-antitrypsin concentrations were observed. The values seem to decrease again premenstrually. Menstrual blood shows very high values, similar to those in blood serum. DISCUSSION
The results of this study on a)-antitrypsin in cervical mucus were obtained from only 10 cycles. However, the preovulatory and postovulatory changes of this protein were uniform and very pronounced. Therefore, it seems permissible to comment on these results.
VOL.
19, No.1, 1968
al-ANTITRYPSIN
97
The preovulatory decrease of acantitrypsin concentrations over a wide range could be interpreted as an effect of increasing estrogen activity. It is conceivable that the decrease in concentration is due to dilution. However, decreased secretion or selective re-resorption should be considered as another possible mechanism. The postovulatory increase and decrease can be interpreted as a progesterone effect. Support for this hypothesis has been obtained from the study of cervical mucus of women under the sequential regimen of hormonal contraception. Figure 6 shows as a
Fig. 6. BBT, spinnbarkeit, fern test, cervical mucus cellularity, mucorrhea, and antitrypsin levels during cycle of patient treated with C-Quens.
al-
SCHUMACHER & PEARL
FERTILITY & STERILITV
typical example the observations made during a cycle under C-Quens* medication. The aI-antitrypsin decreases remarkably during the first 5 days and remains at low values under estrogen administration. The values increase as soon as the combined estrogen-progestational preparation has been taken. High values in this range have been observed also in patients undergoing treatment with combined preparations. The pattern of acantitrypsin changes under the influence of sex hormones appears to be similar to electrophoretic changes of the albumin fraction described recently by Moghissi et al. 9 ,12 However, the question remains as to whether the other serum proteins present in cervical mucus also follow a similar pattern. Little is known about the physiologic function of aI-antitrypsin. If proteolytic enzymes in semen playa role in the process of sperm penetration (a hypothesis which has been presumed by Moghissi et al. IO ), the hormone-dependent decrease and increase of a powerful protease inhibitor could be an important factor in the mechanism of sperm migration. 431 Ashland Ave. Homewood, Ill.
REFERENCES 1. BUNDY, H. F., and MEHL, J. W. Trypsin inhibitors of human serum. II. Isolation of the arinhibitor and its partial characterization. J Bioi Chem 234: 1124, 1959. 2. CLEVE, H. Quantitative immunologische Bestimmung von saurem al-glykoprotein und aI-antitrypsin: Serumkonzentrationen bei gesunden Blutspendern. Klin Wschr 44: 1256, 1966. 3. HEIMBURGER, N., and SCHWICK, G. Die Fibrinagar-Elektrophorese. 1. und II. Mitt. Thromb Diath Haemorrh 7:432, 1962. 4. HEINEN, G. "Immunelektrophoretische Untersuchungen del' weiblichen vaginal und zervikal Sekretion." In Protides of the Biological Fluids (Vol. 12). Elsevier, Amsterdam, 1965, pp. 210-212. 5. KUEPPERS, F., BRISCOE, W. A., and BEARN, A. G. Hereditary deficiency of serum aI-antitrypsin. Science 146:1678, 1964. 6. LAURELL, C.-B., and ERIKSSON, S. The electrophoretic aI-globulin pattern of serum in aI-antitrypsin deficiency. Scand J Clin Lab Invest 15:132, 1963. 7. MANCINI, G., CARBONARA, A. 0., and HEREMANS, J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemist'fy 2:235, 1965. 8. MANCINI, G., VAERMAN, J.-P., CARBONARA, A. 0., and HEREMANS, J. F. "A Singleradial-diffusion Method for the Immunological Quantitation of Proteins." In Protides of the Biological Fluids (Vol. 11). Elsevier, Amsterdam, 1964, pp. 370-373. 9. MOGHISSI, K. S. Cyclic changes of cervical mucus in normal and progestintreated women. Fertil Steril17:663, 1966. 10. MOGHISSI, K. S., DABICH, D., LEVINE, J., and NEUHAUS, O. W. Mechanism of sperm migration. Fertil Steril15:15, 1964. 11. MOGHISSI, K. S., and NEUHAUS, O. W. Composition and properties of human cervical mucus. II. Immunoelectrophoretic studies of the proteins. Amer J Obstet Gynec 83:149, 1962. *Eli Lilly Co., Indianapolis, Ind.
VOL. 19, No.1, 1968
(q -ANTITRYPSIN
99
12. MOGHISSI, K. S., and NEUHAUS, O. VV. Cyclic changes of cervical mucus proteins. ArneI' ] Obstet Gynec 96:91, 1966. 13. SCHULTZE, H. E., and HEREMANS, J. F. Molecular Biology of Human Proteins. Nature and Metabolism of Extracellular Proteins (Vol. 1). Elsevier, Amsterdam, 1966. 14. SCHULTZE, H. E., HEIDE, K., and HAUPT, H. al-Antitrypsin aus Humanserum. Klin Wschr 40:427, 1962. 15. SCHULTZE, H. E., HEIMRl'RGER, N., HEIDE, K., HAUPT, H., STORIKO, L., and SCHWICK, H. C. Preparation and Characterization of arTrypsin Inhibitor and a2-Plasmin Inhibitor of Human Serum. In Proceedings of the 9th Congress of the European Society of Hematology held in Lisbon in 1963. Karger, Basel, 1963, pp. 1315-1320. 16. SCHUMACHER, C. F. B. Unpublished data. 17. SCHUMACHER, C. F. B. Protein analysis of secretions of the female genital tract. ] Reprod M ed (In press). 18. SCHUMACHER, C. F. B., STRAUSS, E. K., and \VIED, C. L. Serum proteins in cervical mucus. Amer] Obstet Gynec 91:1035, 1965. 19. SCHUMACHER, C. F. B., and WIED, C. L. "Semiquantitative Microanalysis of Proteins in Cervical Mucus." In Proceedings of the Vth World Congress on Fertility and Sterility, Stockholm, 1966. International Congress Series, Westin, B., and Wiqvist, N., Eds. Excerpta Medica Foundation, Amsterdam, 1966, p. 975. 20. SCHWICK, H. C. "Quantitative Immunpracipitation zur Bestimmung einzelner Plasmaproteine." In 15th Colloquium Ges Physiol Chern, Mosbach, Westphal, 0., Ed. Springer, Berlin, 1964, pp. 55-67. 2l. SCHWICK, C., and STORIKO, K. "Qualitative and Quantitative Determination of Plasma Proteins by Immunoprecipitation." In Lab Synopsis (Vol. 1). Lloyd Brothers, Cincinnati, 1965, pp. 1-68.
Association of Clinical Endocrinology The Association of Clinical Endocrinology will hold a Symposium on the physiopathology of adipose tissue, entitled "Troisiemes Joumees Internation ales d'Endocrinologie de Marseille," on May 9-12, 1968, in Marseille, France. For further information, please contact the Secretary-General, Prof. AG. J.-L. CODACCIONI, Hopital de la Conception, 144, Rue Saint-Pierre, F.13, Marseille, France.
of human serum allows rough differentiation of albumin, a1- and a2-globulins, fi-globulins, and y-globulins according to their electrophoretic mobility. These fractions, except albumin, are nonhomogeneous. They consist of several subfractions with individual molecular characteristics. High-resolution systems and immunochemical methods permit further differentiation. More than 30 proteins, glycoproteins, and lipoproteins are now well characterized. The major component of a group of glycoproteins and lipoproteins that migrate with the electrophoretic mobility of a1-globulins is a1-antitrypsin. Its synonyms are al-3,5-glycoprotein, au-globulin, and a,-seromucolde de Schultze. 1, 5, 6,13,14 This a1-globulin is a glycoprotein (12% carbohydrate) with a molecular weight of 45,000, which is small as compared with those of other serum proteins. It inhibits trypsin and chymotrypsin and has therefore been named al-antitrypsin. 13 , 14 Its capacity to inhibit trypsin is in the range of 0.25-0.50 mg. trypsin per milligram of a1-antitrypsin. ac antitrypsin also inhibits autoactivated plasmin but not streptokinaseactivated plasmin. 15 The normal range of this protein appears to be between 2 and 5 mg.jml. of serum. 2 ,13,15 The immunoelectrophoretic analysis of cervical mucus indicates in most normal specimens the presence of acantitrypsin (Fig. 1), as previously reported. 18 If developed with antiserum against total human serum, the immunoelectrophoretic pattern shows five precipitin lines. Specific antiserum against purified a1-antitrypsin identifies the precipitin line in the a1-globulin area as a1-antitrypsin.17,18 The other bands indicate the presence of prealbumin, albumin, transferrin, and 7S y-globulin, as previously ELECTROPHORETIC SEPARATION
From the Division of Laboratories and Research, New York State Department of Health, and from the Departments of Obstetrics and Gynecology, and of Biochemistry, Albany Medical College, Albany, N. Y. Presented at the 23rd Annual Meeting of The American Fertility Society, Washington, D. C., Apr. 14--16, 1967. We thank Miss Margarita Feifel for her technical assistance. Supported in part by Grant IROI HD 02682-01 from the U. S. Public Health Service. *Present address: University of Chicago, Department of Obstetrics and Gynecology, The Chicago Lying-in Hospital, Chicago, Ill.
91
92
SCHUMACHER
&
PEARL
FERTILITY
& STERILITY
Fig. 1. Immunoelectrophoresis of cervical mucus.
reported by Moghissi and Neuhaus l l and by Heinen. The trypsin-inhibitor activity of !Xl-antitrypsin in cervical mucus has been established16 by a combined immunoelectrophoretic and enzyme diffusion technic in fibrincontaining agar gel,3 Immunoelectrophoretic technics of this type provide only qualitative information. The quantitative assessments of proteins present in cervical mucus would be highly desirable to gain insight in secretory functions under the influence of hormones and drugs and under pathologic conditions. METHODS
Immunochemical micromethods are the methods of choice to determine concentrations of soluble proteins in cervical mucus specimensY' 19-21 A simple and accurate method of high sensitivity is the single-radial-immunodiffusion technic in agar gel, introduced a few years ago by Mancini et ap· 8 The principles of this method are illustrated in Fig. 2. A commercially available antiserum* against !Xl-antitrypsin is incorporated into a buffered, uniformly thin, agar gellayer.t The cervical mucus is deposited *Hoechst Pharmaceutical Co., Kansas City, Mo. tDetails of the preparation of the gel are being published elsewhere." Ready-to-use "Partigen"-Immunodiffusion Plates are now also available from Hoechst Pharmaceutical Co.
VOL.
19, No.1, 1968
aI-ANTITRYPSIN
93
into wells, which are punched in the gel with stainless-steel well cutters of 3-mm. diameter.* The lXI-antitrypsin diffuses (with other soluble compounds) radially into the antibody-containing agar, forming a growing disc of antigen-antibody precipitates. The growth of the discs is a function
ANTIGEN SOLUTION
Fig. 2. Principles of singleradial-immunodiffusion technic in agar gel.
ANTIBODY' CONTAINING AGAR
_lADIA~~~~;'ON 8J.dFJ m IDE
O-TIME
AFTER MINUTES
AFTER HOURS
of antigen concentration, of time, of antigen-antibody ratio, and of the diffusion coefficient of the diffusing molecules. The diameter of the discs can be measured and related to standard solutions of purified IXl-antitrypsint Figure 3 shows the results of an experiment with standard solutions of known concentrations. The square of the radius of the discs plotted against the concentration of the standard solutions shows almost a straight line in concentrations between 10 and 1000 /Lg. jml. if the precipitates are measured after 72 hr. of diffusion at room temperature. The r2 values of the discs produced by cervical mucus specimens can be evaluated now by this standard curve. Each series of lXI-antitrypsin determinations requires the use of these standard solutions. Specimens and standard solutions in small portions are stored at 20° C. RESULTS
Cervical mucus specimens from normal healthy women during ovulatory cycles and under the influence of hormonal contraceptives have been investigated by means of this immunologic method for determining the IX]antitrypsin content. A typical picture of an ovulatory cycle is shown in Fig. 4. The lXI-antitrypsin concentration decreases from high postmenstrual values to very low values between Days 11 and 13 and increases again remarkably on Day 14. The low values coincide with high spinnbarkeit values, positive Fern test, very low cellularity, mucorrhea, and positive Sims-Huhner tests. According to these parameters and to the increase of *Immunodiffusion Set, LKB-Instruments, Inc., Rockville, Md. tThe purified ",,-antitrypsin was made available by the courtesy of Dr. Schwick, Behringwerke, Marburg, Germany.
94
SCHUMACHER
&
PEARL
FERTILITY
&
STERILITY
basal body temperature, one can assume that ovulation has occurred on about the twelfth day. Figure 5 shows the preovulatory and postovulatory courses of aI-antitrypsin, spinnbarkeit, and basal body temperature (BBT) of 10 patients. In three of these patients, ovulation was induced by administration of gonadotropins. The day of minimum acantitrypsin values, which ranges between Days 10 and 16 of the spontaneous cycles, has been defined in this
2 YtHrs.
,50 100
,500
Fig. 3. Results of experiment using standard solutions of known concentrations in single-radial-immunodiffusion technic. The growth of the precipitin discs over a period of 72 hr. of diffusion is demonstrated.
VOL.
19, No.1, 1968
95
lq -ANTITRYPSIN
graph as Day o. The days prior to the day of minimal aI-antitrypsin values and the following days are arranged in order as minus or plus days. The spinnbarkeit and BBT values are arranged corresponding to their respective aI-antitrypsin values. The BBT curves and other clinical data give evidence for biphasic cycles in all cases and suggest that ovulation did occur. Two of these individuals became pregnant during the cycle studied. Minimum values of aI-antitrypsin concentration and maximum values of spinnbarkeit
5
o
GOO 500 400
300
200 100
Menses
o
3 4 5 6
~~
9 10 1{12 13 14 IfilS 17 18 19 20 2122
000 00,0
..
a 2425 262728C~
···Qh; .·cdaJj
···fMMUNOPREClPITATES oFD(;"ANTI TRYPSiN;>: Fig. 4. The aI-antitrypsin content of cervical mucus, fem test, presence of cellularity, occurrence of mucorrhea, Sims-Huhner test results, spinnbarkeit, and BBT during ovulatory menstrual cycle.
96
SCHUMACHER
& .PEARL
F'ERTILITY
& STERILl'ry
are quite well correlated and coincide with the last day prior to the stepwise increase of basal body temperature. Therefore, one may assume that ovulation occurred on Day 0 or on Day +1 in some cases. If one compares these three parameters, it appears that the al-antitrypsin values show only "F 99
Basal bady temperature 98 975 97
C:Oj
Spinnbarkeit
10
5
o
___
l-~../
I'o/ml
700
600 500 400 300
200
0(,
-Antitrypsin
100
Fig. 5. Pre- and postovulatory courses of al-antitrypsin, spinnbarkeit, and BBT in 10 patients. G indicates cycles in which ovulation was induced by administration of gonadotropins. Arrow indicates day of minimum al-antitrypsin concentration.
little increase during the first 48 hr. after the minimum on Day 0, while the changes in the spinnbarkeit values and in the BBT values are more pronounced. From the second or third dayan, significantly higher al-antitrypsin concentrations were observed. The values seem to decrease again premenstrually. Menstrual blood shows very high values, similar to those in blood serum. DISCUSSION
The results of this study on a)-antitrypsin in cervical mucus were obtained from only 10 cycles. However, the preovulatory and postovulatory changes of this protein were uniform and very pronounced. Therefore, it seems permissible to comment on these results.
VOL.
19, No.1, 1968
al-ANTITRYPSIN
97
The preovulatory decrease of acantitrypsin concentrations over a wide range could be interpreted as an effect of increasing estrogen activity. It is conceivable that the decrease in concentration is due to dilution. However, decreased secretion or selective re-resorption should be considered as another possible mechanism. The postovulatory increase and decrease can be interpreted as a progesterone effect. Support for this hypothesis has been obtained from the study of cervical mucus of women under the sequential regimen of hormonal contraception. Figure 6 shows as a
Fig. 6. BBT, spinnbarkeit, fern test, cervical mucus cellularity, mucorrhea, and antitrypsin levels during cycle of patient treated with C-Quens.
al-
SCHUMACHER & PEARL
FERTILITY & STERILITV
typical example the observations made during a cycle under C-Quens* medication. The aI-antitrypsin decreases remarkably during the first 5 days and remains at low values under estrogen administration. The values increase as soon as the combined estrogen-progestational preparation has been taken. High values in this range have been observed also in patients undergoing treatment with combined preparations. The pattern of acantitrypsin changes under the influence of sex hormones appears to be similar to electrophoretic changes of the albumin fraction described recently by Moghissi et al. 9 ,12 However, the question remains as to whether the other serum proteins present in cervical mucus also follow a similar pattern. Little is known about the physiologic function of aI-antitrypsin. If proteolytic enzymes in semen playa role in the process of sperm penetration (a hypothesis which has been presumed by Moghissi et al. IO ), the hormone-dependent decrease and increase of a powerful protease inhibitor could be an important factor in the mechanism of sperm migration. 431 Ashland Ave. Homewood, Ill.
REFERENCES 1. BUNDY, H. F., and MEHL, J. W. Trypsin inhibitors of human serum. II. Isolation of the arinhibitor and its partial characterization. J Bioi Chem 234: 1124, 1959. 2. CLEVE, H. Quantitative immunologische Bestimmung von saurem al-glykoprotein und aI-antitrypsin: Serumkonzentrationen bei gesunden Blutspendern. Klin Wschr 44: 1256, 1966. 3. HEIMBURGER, N., and SCHWICK, G. Die Fibrinagar-Elektrophorese. 1. und II. Mitt. Thromb Diath Haemorrh 7:432, 1962. 4. HEINEN, G. "Immunelektrophoretische Untersuchungen del' weiblichen vaginal und zervikal Sekretion." In Protides of the Biological Fluids (Vol. 12). Elsevier, Amsterdam, 1965, pp. 210-212. 5. KUEPPERS, F., BRISCOE, W. A., and BEARN, A. G. Hereditary deficiency of serum aI-antitrypsin. Science 146:1678, 1964. 6. LAURELL, C.-B., and ERIKSSON, S. The electrophoretic aI-globulin pattern of serum in aI-antitrypsin deficiency. Scand J Clin Lab Invest 15:132, 1963. 7. MANCINI, G., CARBONARA, A. 0., and HEREMANS, J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemist'fy 2:235, 1965. 8. MANCINI, G., VAERMAN, J.-P., CARBONARA, A. 0., and HEREMANS, J. F. "A Singleradial-diffusion Method for the Immunological Quantitation of Proteins." In Protides of the Biological Fluids (Vol. 11). Elsevier, Amsterdam, 1964, pp. 370-373. 9. MOGHISSI, K. S. Cyclic changes of cervical mucus in normal and progestintreated women. Fertil Steril17:663, 1966. 10. MOGHISSI, K. S., DABICH, D., LEVINE, J., and NEUHAUS, O. W. Mechanism of sperm migration. Fertil Steril15:15, 1964. 11. MOGHISSI, K. S., and NEUHAUS, O. W. Composition and properties of human cervical mucus. II. Immunoelectrophoretic studies of the proteins. Amer J Obstet Gynec 83:149, 1962. *Eli Lilly Co., Indianapolis, Ind.
VOL. 19, No.1, 1968
(q -ANTITRYPSIN
99
12. MOGHISSI, K. S., and NEUHAUS, O. VV. Cyclic changes of cervical mucus proteins. ArneI' ] Obstet Gynec 96:91, 1966. 13. SCHULTZE, H. E., and HEREMANS, J. F. Molecular Biology of Human Proteins. Nature and Metabolism of Extracellular Proteins (Vol. 1). Elsevier, Amsterdam, 1966. 14. SCHULTZE, H. E., HEIDE, K., and HAUPT, H. al-Antitrypsin aus Humanserum. Klin Wschr 40:427, 1962. 15. SCHULTZE, H. E., HEIMRl'RGER, N., HEIDE, K., HAUPT, H., STORIKO, L., and SCHWICK, H. C. Preparation and Characterization of arTrypsin Inhibitor and a2-Plasmin Inhibitor of Human Serum. In Proceedings of the 9th Congress of the European Society of Hematology held in Lisbon in 1963. Karger, Basel, 1963, pp. 1315-1320. 16. SCHUMACHER, C. F. B. Unpublished data. 17. SCHUMACHER, C. F. B. Protein analysis of secretions of the female genital tract. ] Reprod M ed (In press). 18. SCHUMACHER, C. F. B., STRAUSS, E. K., and \VIED, C. L. Serum proteins in cervical mucus. Amer] Obstet Gynec 91:1035, 1965. 19. SCHUMACHER, C. F. B., and WIED, C. L. "Semiquantitative Microanalysis of Proteins in Cervical Mucus." In Proceedings of the Vth World Congress on Fertility and Sterility, Stockholm, 1966. International Congress Series, Westin, B., and Wiqvist, N., Eds. Excerpta Medica Foundation, Amsterdam, 1966, p. 975. 20. SCHWICK, H. C. "Quantitative Immunpracipitation zur Bestimmung einzelner Plasmaproteine." In 15th Colloquium Ges Physiol Chern, Mosbach, Westphal, 0., Ed. Springer, Berlin, 1964, pp. 55-67. 2l. SCHWICK, C., and STORIKO, K. "Qualitative and Quantitative Determination of Plasma Proteins by Immunoprecipitation." In Lab Synopsis (Vol. 1). Lloyd Brothers, Cincinnati, 1965, pp. 1-68.
Association of Clinical Endocrinology The Association of Clinical Endocrinology will hold a Symposium on the physiopathology of adipose tissue, entitled "Troisiemes Joumees Internation ales d'Endocrinologie de Marseille," on May 9-12, 1968, in Marseille, France. For further information, please contact the Secretary-General, Prof. AG. J.-L. CODACCIONI, Hopital de la Conception, 144, Rue Saint-Pierre, F.13, Marseille, France.