Effects of atrial natriuretic peptide on rat ovarian granulosa cell steroidogenesis in vitro.

PROBLEM: The localization and biological effects of atrial natriuretic peptide (ANP) are not limited to cardiac tissue but extend to a number of extra-atrial tissues and organs, including the ovary. The objective of the present study was to determine the effects of ANP on granulosa cell steroidogenesis. Hence, the direct effects of ANP on the production of progesterone, 20 alpha-hydroxpregn-4-en-3-one (20 alpha-OH-P), and estrogen by undifferentiated and differentiated rat ovarian granulosa cells were examined in vitro. METHOD: Undifferentiated granulosa cells obtained from the ovaries of diethylstilbestrol-primed, immature rats were treated with increasing doses (10(-12) to 10(-6) M) of rANP for 48 h. RESULTS: ANP evoked a two- and threefold increase in progesterone and 20 alpha-OH-P production relative to untreated controls, respectively. Increasing doses of ANP in combination with porcine FSH (125 ng/well) resulted in a biphasic response in progesterone production above FSH-treated controls. Specifically, a maximal inhibition of 35% in progesterone production was achieved at 10(-9) M ANP, followed by a stimulation to levels comparable with FSH-stimulated controls at higher doses examined. Increasing doses of ANP evoked a twofold increase in FSH-stimulated 20 alpha-OH-P production over respective controls. Following pretreatment of granulosa cells with FSH for 48 h to evoke differentiation, ANP caused a significant dose-dependent inhibition in basal progesterone production that resulted in a 76% suppression at the highest dose examined. In contrast, ANP evoked a 2.8-fold increase in 20 alpha-OH-P production when compared with controls. Finally, in FSH-stimulated differentiated granulosa cells, ANP evoked a threefold increase in progesterone production and a 65% inhibition in 20 alpha-OH-P production. ANP exerted no significant effects on estrogen production by either undifferentiated or differentiated granulosa cells in the presence or absence of FSH. CONCLUSIONS: These data demonstrate that ANP can modulate directly progestin steroidogenesis in both undifferentiated and differentiated rat ovarian granulosa cells in vitro and, therefore, may play an important role in granulosa cell differentiation and follicular maturation.

Antigenic differences between the endometrium of women with and without endometriosis.

Serum and peritoneal fluid from 12 women with endometriosis, 4 women with uterine leiomyomata and 6 fertile women without endometriosis (controls) and serum from 4 women with adenomyosis were tested with a passive hemagglutination assay for antibodies against endometrium from all the controls, 8 patients with endometriosis and all patients with uterine leiomyomata and from implants from 8 patients with endometriosis. Serum antibody titers in patients with endometriosis or leiomyomata were significantly higher against endometrial or implant antigens from patients with endometriosis and 2 patients with leiomyomata than those against the controls' endometrium. Peritoneal fluid endometrial antibody titers failed to reflect these antigenic differences. Controls and patients with adenomyosis had low titers of endometrial antibodies in their serum or peritoneal fluid. Antigenic differences appear to exist between the endometrium of patients with endometriosis and that of controls.

Influence of the menstrual cycle on systemic diseases.

Physiological changes associated with the menstrual cycle influence the clinical course of some diseases such as bronchial asthma, allergies, anaphylaxis, epilepsy, migraine, dermatoses, and porphyria. Hormonal manipulation can be beneficial in some patients.

White cell subpopulations in autoimmune infertile men and their wives with cytotoxic sperm antibodies.

Sixty six infertile men with cytotoxic sperm antibodies had fewer and lower percentages of OKT4-positive helper/inducer T lymphocytes p = 0.004 and 0.003, respectively), than the fertile (n = 14) and infertile (n = 12) antibody-negative controls. Ratio of OKT4/OKT8 was decreased (p less than 0.01), while numbers of OKDR and BD-Leu12 positive predominantly B lymphocytes were increased (p less than 0.0001) in infertile men with sperm antibodies. T lymphocytes positive for OKT3, OKT11 and OKT4 and the OKT4/OKT8 ratio were decreased in infertile women with sperm antibodies, while B lymphocytes positive for OKDR and BD-Leu12 were increased in infertile women with or without sperm antibodies (p less than 0.0001, versus fertile controls). Lymphocyte responses to phytohemagglutinin (PHA), concanavalin A (ConA) and pokeweed mitogen (PWM), however, were similar in 20 fertile and 136 infertile couples. Lymphocytes from infertile men with sperm antibodies had an enhanced stimulatory response to autologous sperm and seminal plasma. It is concluded that sperm antigens from autoimmune infertile men stimulate heightened immune responses to sperm antigens and altered distribution of white cell populations in both partners.

Bilateral fallopian tubal polyps: radiologic and pathologic correlation.

A case of bilateral fallopian tube filling defects found on hysterosalpingography performed on a patient for infertility work-up is presented. Repeat hysterosalpingography performed 4 years later showed no change. Microsurgical resection revealed the defects to be fallopian tubal polyps of endometrial origin.

Time-related decline in sperm motility patterns in men with cytotoxic antibodies.

Sperm motility patterns in semen from 10 fertile nonautoimmune men (fertile control group) and 33 infertile men with various titers of cytotoxic sperm antibodies in their seminal plasma (group 1: titers less than or equal to 16, n = 6; group 2: titers 64 to 512, n = 12; group 3: titers greater than or equal to 1024, n = 15) were evaluated every 2 hours for 12 hours and finally at 24 hours. A significant decline in sperm swimming speed and linearity was observed at 6 hours in semen from 27 infertile men with sperm antibodies. Beginning at 8 hours, semen from sperm antibody-positive men in group 2 showed a significant decline in percentage motile sperm (p less than 0.001) compared to the fertile controls. The percentage motility in semen of donors in groups 1 and 3 was significantly lower than that in semen of fertile donors at 10 hours (p less than 0.05), 12 hours (p less than 0.01), and 24 hours (p less than 0.001). The mean velocity in groups 2 and 3 was significantly less than that in fertile controls at 10 and 12 hours (p less than 0.05). The linearity of sperm motility started to decline 4 hours after semen samples were obtained from sperm antibody-positive groups 2 and 3 in contrast to sperm antibody-negative fertile or infertile groups (p less than 0.05). It is concluded that the presence of cytotoxic sperm antibodies in the seminal plasma adversely affects sperm linearity as early as 4 hours after semen collection.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunological aspects of surgically induced experimental endometriosis: variation in response to therapy.

Elevated endometrial antibody titers were detected in serum and peritoneal fluid of rabbits with experimentally induced endometriosis. Surgical extirpation of implants or suppression with gonadotropin-releasing hormone agonist resulted in decreased endometrial antibody titers, whereas the antibody titers in untreated rabbits with endometriosis increased significantly. Endometrial implants and normal endometrial tissue had similar proteins by polyacrylamide gel electrophoresis. However, the serum and peritoneal fluid from rabbits with experimentally induced endometriosis had gamma G immunoglobulin antibodies to an endometrial protein with molecular weight of approximately 40 kD. These antibodies were absent in rabbits without endometriosis. Isolation of endometrial antigens eliciting the humoral immune response in endometriosis may aid in the development of a specific antibody marker for endometriosis.

Endometrial antigens involved in the autoimmunity of endometriosis.

Serum and peritoneal fluid from five fertile women without endometriosis and serum (n = 23) and peritoneal fluid (n = 12) from infertile women with endometriosis were tested for the presence of antibodies against endometrial tissue antigens by a Western blot analysis. Antigens with molecular weights (MW) of 19, 31, 38, and 42 kd reacted with antibodies in the serum and peritoneal fluid from both fertile and infertile women. Antibodies in 20 of 23 (87%) sera and all 12 (100%) peritoneal fluid samples from endometriosis patients reacted against endometrial antigens with molecular weights (MW) of 26 kd and/or 34 kd. Serum from 10 patients (43%) and peritoneal fluid from 6 patients (50%) also had antibodies to an endometrial antigen with MW of 21.5 kd. Reactivity to other endometrial antigens with MW 16, 24, 48, and 75 kd was also noted in patients with endometriosis. Antibodies in the serum and peritoneal fluid from fertile women failed to react against these antigens. It is concluded that the humoral and local endometrial autoimmunity detected in patients with endometriosis is primarily directed against antigens with MW of 26 and 34 kd.

Studies on sperm survival and motility in the presence of cytotoxic sperm antibodies.

The effect of cytotoxic sperm antibodies and native complement in the serum and secretions from 40 fertile and 93 infertile couples on in vitro sperm survival and motion characteristics was studied. Sperm survival in vitro was unaffected by sera from fertile and infertile subjects without cytotoxic sperm antibodies and from infertile men with antibodies to control but not to autologous sperm. Sperm survival was reduced (P less than .001) by sera from infertile men with antibodies to autologous sperm or to antologous and control sperm and from women with cytotoxic antibodies to sperm from both. Sera from fertile couples without sperm antibodies enhanced sperm swimming speed and motility index (P less than .0001). Sera from infertile women with or without cytotoxic sperm antibodies did not affect sperm motility. Sperm survival and motility were reduced by seminal plasma from infertile men with cytotoxic antibodies to autologous and/or control sperm. Seminal plasma from fertile men enhanced sperm survival. Cervical mucus from infertile women with antibodies to autoimmune husbands' sperm or to husbands' and control sperm inhibited sperm motion, whereas cervical mucus from infertile women without sperm antibodies and women with antibodies to control sperm failed to have any effect. It is concluded that cytotoxic sperm antibodies developed through exposure to sperm antigens in autoimmune infertile men decrease in vitro sperm survival and/or motility.

Special antigens on sperm from autoimmune infertile men.

Sera from three fertile men and four infertile men without sperm antibodies, 17 infertile men with sperm antibodies in serum and seminal plasma (S.P.), and 25 infertile men with sperm antibodies in S.P. were tested by Western Blot analysis against sperm membrane extracts and S.P. from fertile nonautoimmune men and infertile autoimmune men. Sera from fertile men reacted against common antigens with molecular weights (MW) of 28, 38, 48, 60, and 68 kD present on sperm from autoimmune and nonautoimmune men and special antigen of MW 76 kD on the sperm of fertile men. Sera from 15 of 17 (88%) autoimmune infertile men with sperm antibodies in serum and S.P. detected special antigens with MW of 58 kD (sera reactivity in 47% of these men), 43kD (in 29%), 30 kD (in 24%), 35 kD (in 18%), 52 kD (in 12%), 41 kD (in 6%), and 71 kD (in 6%) on the sperm of autoimmune men in addition to the common antigens. Sera from 15 of 25 (60%) men with sperm antibodies in their S.P. showed reactivity to special antigens with MW 52 kD (in 20%), 35 kD (in 16%), 41 kD (in 16%), 58 kD (in 8%), 70/71 kD (in 8%), 30 kD (in 8%), and 56 kD (in 4%). Sera from 18 of 42 (43%) infertile men with sperm antibodies also detected special antigens of MW 26, 46, and 76 kD present only in fertile men's sperm. Sera from only 15 of 42 (36%) autoimmune infertile men reacted against special antigens with MW 17, 20, 23, 30, 43, and 58 kD in the seminal plasma of autoimmune infertile men.(ABSTRACT TRUNCATED AT 250 WORDS)

A computerized postcoital test sperm motility: comparison with clinical postcoital test and correlations with sperm antibodies.

Sixty-five infertile women had a conventional postcoital test (PCT), a computerized postcoital test (cPCT), and sperm antibody testing. Twenty-four women had good cervical mucus and good PCT sperm motility (group 1), 23 had poor cervical mucus and poor PCT sperm motility (group 2), and 18 had good cervical mucus but poor PCT sperm motility (group 3). The percentage of motile sperm, mean linearity, and the motility index of sperm by cPCT also were decreased in groups 2 and 3 (p less than 0.001) in contrast to group 1. A reduced PCT sperm count was significantly associated with positive titers of antibodies to autologous sperm in the husbands' serum, whereas a reduced PCT motility correlated with high titers of cytotoxic antibodies to husbands' sperm in the wives' serum and cervical mucus. An increased percentage of vibratory sperm at PCT correlated with elevated titers of cytotoxic antibodies to husbands' sperm in the wives' serum and cervical mucus, and hemagglutinating (r = 0.44; p less than 0.001) and immunofluorescent IgA antibodies to husbands' sperm (r = 0.47; p less than 0.001) in the cervical mucus. Mean swimming speed of sperm by cPCT correlated inversely with cytotoxic and hemagglutinating antibody titers to husbands' sperm, and immunobead-binding IgM and immunofluorescent IgG, IgA, and IgM (r = 0.52; p less than 0.001) antibodies to sperm in the seminal plasma. Motility indices correlated inversely with cytotoxic antibody titers to husbands' sperm in the wives' serum, and hemagglutinating antibody titers to husbands' sperm in cervical mucus. The predictive values of PCT and cPCT for the presence of cytotoxic and immunofluorescent IgA antibodies to autoimmune husbands' sperm were 76% and 71%, respectively, in the serum and 85% and 75%, respectively, in the cervical mucus of the wives. The predictive value of PCT and cPCT for immunobead-binding and immunofluorescent IgM antibodies to sperm in the wives' serum was 71%. Computerized PCT measures more sperm characteristics than PCT, although it is in general agreement with PCT.

Efficacy of sperm wash media in improving sperm motility.

Spermatozoa from 15 fertile men were washed with Ham's F10 and incubated with two commercially available sperm nutrient media for 2, 4, 6, and 24 h. Both sperm capacitation medium (Irvine Scientific Co., Santa Ana, CA) and Pro-ception (Milex Products, Inc., Chicago, IL) proved to be capable of improving sperm motion characteristics. These media may be used for incubating sperm for intrauterine insemination or for in vitro fertilization.

Concurrence of the Holt-Oram syndrome and the Rokitansky-Kuster-Hauser syndrome. A case report.

A 15-year-old white woman, karyotype 46,XX, had an absent vagina and uterus (Rokitansky-Kuster-Hauser syndrome) as well as an atrial septal defect and transphalangeal thumb (Holt-Oram syndrome). The concurrence of the above two syndromes has not been reported on before.

Cytotoxic sperm antibodies and in vitro fertilization of mature oocytes: a preliminary report.

The fertilization rates of mature oocytes during in vitro fertilization and embryo transfer (IVF-ET) using fetal cord serum-supplemented insemination media were greater than or equal to 57% for five infertile couples without sperm antibodies (group 1). But they were less than or equal to 50% for four of nine infertile couples (group 2) with cytotoxic sperm antibodies in both partners (n = 6) or the husband alone (n = 3). Two women in group 1 were successful in achieving normal, full-term pregnancies with the delivery of normal infants (chi2 = 4.2, P less than 0.05, by chi-square analysis). One of them consistently tested negative for sperm antibodies, while her husband was previously treated with antibiotics for infection and transient sperm antibodies in the seminal plasma. Subsequently, antibody titers in the husband were in the normal range when the successful IVF-ET was performed. One woman in group 2, with antibodies to her autoimmune husband's sperm but not control sperm and with a long-standing poor postcoital test sperm motility, conceived through artificial insemination with donor sperm (AID) after failing to conceive with her husband through IVF-ET. These data suggest that the presence of cytotoxic sperm antibodies in the serum and/or secretions of both partners reduces the rates of fertilization of mature oocytes in spite of using fetal cord serum in the IVF media. Pregnancy achievement is impaired in this group.

Interleukin-1: a possible role in the infertility associated with endometriosis.

Increased numbers of activated macrophages are associated with mild endometriosis. Interleukin-1 (IL-1) is a protein produced by macrophages and is believed to be a primary mediator of host responses. IL-1 induces prostaglandin and fibrinogen synthesis and stimulates fibroblast proliferation. This study was undertaken to evaluate the role of IL-1 in the infertility associated with mild endometriosis. Peritoneal fluid (PF) was obtained at laparoscopy from 11 patients with minimal or mild endometriosis and from 7 women undergoing tubal ligation. Peritoneal macrophages were isolated and cultured for 24 hours. Peritoneal and macrophage culture fluids were studied for IL-1 activity, which was measured with the EL-4 assay. IL-1 activity was present in the PF of 10 of the 11 patients with endometriosis and 11 of the 11 macrophage culture fluids and was absent in the PF and macrophage culture fluid of the tubal ligation patients. The effect of recombinant alpha-IL-1 on the in vitro growth of 2-cell mouse embryos was also studied. IL-1 in concentrations similar to those present in the PF (greater than 1 U/ml) was toxic to mouse embryo development. We conclude that the IL-1 may play a role in the infertility associated with endometriosis.

Sperm antibodies and human leukocyte antigens in couples with early spontaneous abortions.

Twenty-six couples with two or more early spontaneous abortions and 53 couples with recently proven fertility (control group) were typed for human leukocyte antigens (HLA) of A and B loci and screened for the presence of cytotoxic and hemagglutinating antibodies to sperm. Eighteen other couples with two or more abortions were tested for sperm antibodies alone. Both partners in 24 of 44 (54%) aborting couples were positive (titers of greater than or equal to 32), whereas the control group had sperm antibodies in the negative range (titers of 0 to 16; P less than .0001). Thirty-one husbands (70%) and 24 wives (54%) were positive for sperm antibodies in the study group. Sharing of HLA-A and/or -B antigens between partners was similar in the control and study groups. Fourteen of 26 (54%) couples with two or more abortions did not share HLA-A and/or -B antigens, in contrast to 14 of 53 (26%) in the control fertile group (P less than .02). Antigenic frequencies of HLA-B7 singly and in combination with HLA-B35 were increased in females in the aborting but not in the fertile control group (P less than .001). The combined incidence of HLA-B7, -B8, and -B35 in both partners was significantly higher in the aborting couples as contrasted with the control group. It is suggested that the presence of sperm antibodies is associated with early pregnancy wastage. Histocompatibility antigens B7 and B35 may play a role through their association with sperm antibodies and early spontaneous abortions.

Sperm and seminal plasma antigens from autoimmune men induce immunological infertility.

Adult male rabbits were immunized with normal saline (controls), sperm extracts from 2 autoimmune men, seminal plasmas from the same autoimmune men, sperm extract from a fertile nonautoimmune man, and seminal plasma from the same fertile nonautoimmune man. All the sperm donors were free from infections. Rabbits immunized with fertile men's sperm extract and seminal plasma had significantly elevated postimmunization hemagglutinating but not cytotoxic sperm antibodies and reproduced normally. Rabbits immunized with autoimmune men's sperm and seminal plasma antigens developed high titers of cytotoxic and hemagglutinating sperm antibodies in their serum and seminal plasma and their reproduction was markedly reduced. Their sera and seminal plasma reduced motility of sperm from a normal donor. The immune responses were confirmed by electron microscopic immunocytochemistry. This technique revealed membrane-bound endogenous IgG on sperm from only those rabbits immunized with sperm extracts from autoimmune men. These antisera reacted against a protein in the 58,000 D range; antisera to fertile man's sperm extract reacted against three proteins with molecular weights of 15,000, 18,400, 25,000, and 44,000 D, as judged by Western blot. Rabbit antisera to seminal plasma from autoimmune men reacted against several proteins; additionally, it detected two proteins with 43,000 and 68,000 molecular weight detected by antiserum to fertile man's seminal plasma. Sperm and seminal plasma antigens from autoimmune men are different in their elicited immunogenic responses from those of fertile nonautoimmune men. These responses are relevant to infertility.

Motion characteristics of spermatozoa from men with cytotoxic sperm antibodies.

Semen samples from 55 fertile nonautoimmune and 44 infertile sperm autoimmune men were evaluated by computerized sperm cell motion analysis. Sperm counts (mean +/- SEM, 59.6 +/- 10.3 X 10(6) per ml), motility (39.0 +/- 4.6%), mean swimming speed (micron/sec, 26.5 +/- 0.9), mean linearity (straight line distance of the cell track divided by the actual track length and multiplied by 10, 6.5 +/- 0.2), and motility index (% motility X mean speed, 10.7 +/- 1.4) in 23 men with significant titers of cytotoxic sperm antibodies in their serum and seminal plasma were less (p less than 0.0001) than those in the fertile controls. However, these parameters were comparable in 18 men with sperm antibodies in their seminal plasma but not in their serum, and the control group. Infertile men with serum cytotoxic sperm antibodies had more sperm cells swimming at 11-30 micron/sec, and fewer moving at 31 micron or higher; this was in contrast to results obtained from fertile men (p less than 0.05). The percentages of sperm cells moving at 21-30 micron/sec were increased, while those moving at 51-60 microns/sec were decreased in men with seminal plasma sperm antibodies, versus controls. Spermatozoa with low linearities (less than or equal to 6) were higher (p less than 0.05) in men with serum and seminal plasma cytotoxic sperm antibodies than in the fertile group.(ABSTRACT TRUNCATED AT 250 WORDS)

An endometrial antibody assay in the clinical diagnosis and management of endometriosis.

Coded serum samples from 11 normal fertile men and 17 fertile women without endometriosis (control groups) and 41 women with endometriosis were tested blindly for the presence of endometrial antibodies by use of a passive hemagglutination assay. Endometrial antibodies were either absent or present in low baseline titers in the serum samples from the control group. In contrast, 17 of the 23 (74%) patients with untreated endometriosis had elevated titers of endometrial antibodies in their serum. Of the 18 patients treated with danazol, endometrial antibodies were absent in 7 women who showed a good response at repeat laparoscopy, whereas 4 of 5 women with a poor response had significantly positive titers of antibodies. Six patients treated with danazol did not have repeat laparoscopy, but were found to have endometrial antibody titers in the baseline control range. Endometrial antibody titers in women with a good response to danazol were significantly lower than those in women with untreated endometriosis or with a poor response to danazol (P = 0.003). No correlation was observed between the antibody titers and the stage of endometriosis. The results suggest that the assay for serum endometrial antibodies may prove to be a clinically useful, noninvasive aid in the diagnosis of endometriosis. Sequential determination of endometrial antibody titers may be helpful in assessing the efficacy of pharmacologic therapy for endometriosis and evaluating the cases of patients with possible recurrence of the disease.

A computerized sperm motion analysis [published erraturm appears in Fertil Steril 1986 Nov;46(5):980].

Semen samples from 35 fertile men and husbands of 32 infertile couples were evaluated with the use of a rapid computerized sperm motion analysis (CSA). The counts and percentages of motile sperm measured by routine semen analysis (RSA) and CSA were comparable in both groups. In CSA, mean values of swimming speed (microns per second), linearity, and motility index of sperm in the fertile men were significantly higher than those in the infertile group. At 3 hours, the number of sperm moving at a speed of 21 to 30 microns/second were higher in the infertile group than in the fertile group. In contrast, the number of sperm moving at a speed of 31 to 50 microns/second and higher was increased in the fertile group over the infertile group. Good correlations were obtained between RSA and CSA sperm counts and percentages of progressively motile sperm. Sperm counts, percent motility, indices of motility, and mean sperm speed obtained by two observers with the use of CSA correlated well. It seems that the sperm speed and the motility index remain unaltered in repeat semen analyses of fertile men. It is concluded that a computerized analysis of sperm swimming speed is a reliable and rapid mode for evaluating semen samples and offers more discriminatory sperm motion characteristics than the RSA.