Biomedical subjects
Publications and source records attributed to B Lunenfeld.
Human menopausal gonadotropins were administered in multiple repetitive cycles for ovulation induction. Ovarian response, as judged by peak E2 levels and the day of hCG administration, remained similar in 3-12 immediately successive cycles. We conclude that hMG can be administered in multiple successive cycles without clinically impairing ovarian response.
The response to a rapid intravenous administration of adrenocorticotropin (ACTH, cortrosyn) was compared in 24 unexplained infertile women and 13 fertile women. There was no significant difference in the response to a rapid ACTH stimulation as expressed by the slope of testosterone (T) response in both groups. However, the individual and mean T values prior and following the stimulation were significantly higher in the infertile group (P less than 0.001; P less than 0.005, respectively). There was no significant difference in individual values as well as in mean basal values or in mean values of 17-hydroxyprogesterone (17-OHP) after the stimulation between the two study groups (P less than 0.3; P less than 0.9, respectively). There was no significant difference in individual values as well as in mean values of dehydroepiadnrosterone (DHEA) before or 90 min after the stimulation between the two study groups (P less than 0.1; P less than 0.2, respectively). It can therefore be concluded that the elevated basal testosterone values in the infertile group originated from the ovary. Despite the fact that no attempt was made to reduce androgen values 7 (46.66%) of 15 infertile women who were available for follow-up and treatment conceived following ovulation induction therapy.
Estradiol receptor (RE2) levels were measured in normal (n = 22) and pathological (n = 14) human endometria and in various segments and tissues of the same uteri (n = 8). The highest total estradiol levels (TRE2) were found in midcycle normal cyclic endometria and the lowest in the secretory phase of normal cyclic endometria and in normal postmenopausal endometria (the TRE2 levels in the latter two groups were within similar ranges). The highest nuclear RE2 levels were found in proliferative-phase normal cyclic endometria. TRE2 levels in pathological postmenopausal endometria were high, in the range of those found in proliferative normal cyclic endometria, but the nuclear RE2 levels were low as compared with those in normal proliferative endometria. Examination of the distribution of the TRE2 levels in various segments and tissues of normal proliferative uteri revealed similar trends in all cases as follows: the highest levels were measured in the functional endometrium (fundus) and the lowest in the myometrium (fundus). Basal endometrial TRE2 levels decreased along the longitudinal axis from fundus to isthmus. In 4 cases, TRE2 levels in leiomyoma tissues were higher than those in the corresponding myometrium. These results indicate the possibility of a differing regulating mechanism of the RE2 in various segments or tissues of the human uterus.
Cytosolic and nuclear progesterone receptor levels (RPC, RPN) were measured in normal and in pathological human endometria after single-dose progesterone injection 1-3 h before tissue collection (test subjects) and the results compared to those obtained in controls (no progesterone treatment). In test cases of late proliferative or midcycle normal endometria and in estrogen-primed normal postmenopausal endometria, RPN levels were higher and RPC levels were lower compared with those obtained in control samples. In test cases of postmenopausal pathological endometria, histologically normal endometria of women with postmenopausal bleeding and in one case of endometrial polyp, RPN levels were also higher and RPC levels lower than those in the respective controls, but the change of distribution of the RP (cytosol/nuclear extract) was significantly lower than in normal endometrium.
The anti-estrogenic effect of clomiphene citrate (CC) and tamoxifen (TMX) on cervical mucus was evaluated in a prospective crossover study. Ten women underwent randomized alternate cyclical treatment with either 100 mg CC, 40 mg TMX daily, or with placebo. The effect of CC, TMX and placebo on serum estradiol (E2), cervical mucus secretion and on the development of ovarian follicles was evaluated. Compared with placebo, treatment with CC and TMX significantly increased the number of mature ovarian follicles on the day of assumed ovulation (p less than 0.05), elevated E2 secretion (p less than 0.05) and decreased cervical score (p less than 0.05). It can be concluded that anti-estrogenic agents reduced the secretion of cervical mucus. However, in most cases, the excessive E2 production due to multiple follicular growth overcomes the anti-estrogenic effect.
We assessed the hypothesis that due to variations in the conformation of the progesterone receptor induced by the antiprogestin RU38486 compared to the progestin ORG 2058, differences may result in the association of the receptor with some of the chromatin components. The physical properties of the receptor-bound chromatin fragments released by micrococcal nuclease digestion were characterized by sucrose gradient sedimentation and by gel filtration on Agarose A-1.5m or Agarose A-5m columns. The nuclear fraction was isolated from T47D cells previously exposed to 0.1 microM [3H]RU38486 or 0.1 microM [3H]ORG 2058. Micrococcal nuclease digestion solubilized two receptor forms sedimenting at 4.4 S and 6.3 S for the antiprogestin bound receptor and only one receptor at 4.4 S for the progestin ligated receptor. High-salt buffer dissociated either the antiprogestin or the progestin-bound receptor to smaller receptor forms sedimenting at 3.5 S. Chemical cross-linking with the cross-linker 2-iminothiolane of the micrococcal nuclease solubilized receptor forms resulted in 6.7-S and 4.4-S forms sedimenting on 0.4 M KCl gradients for the antiprogestin and progestin ligated receptors, respectively. Stokes radii of 7.3 nm and 6.4 nm were determined by gel filtration in 0.4 M KCl for the 6.7-S and the 4.4-S receptor forms, respectively. Using the sedimentation coefficient and the Stokes radius, molecular weights of 202,000 and 116,000 were calculated for the antiprogestin and progestin ligated receptors. We conclude that the micrococcal nuclease solubilized antiprogestin ligated receptor is associated with additional or different chromatin components compared to the progestin bound receptor.
The influence of four different copper-containing intrauterine devices (IUD) (Nova-T, Multiload, Fincoid 250 and copper-T 250) on the penetration of human sperm into bovine cervical mucus were assessed. Pooled samples of predetermined sperm concentration were suspended in Earl's medium in which a copper IUD was previously incubated for periods between one hour to fourteen days. The mean copper concentration was determined for each of the medium containing IUD and was found to be between 284 +/- 93 micrograms/100 ml to 392 +/- 138 micrograms/100ml. While there was no adverse effect on sperm motility by the copper-containing medium, there was a significant reduction in the number of sperm penetrated into the bovine cervical mucus as compared to the penetration of sperm suspended in pure Earl's medium. It therefore seems that the influence of copper on sperm penetration might be by an effect on the environment or spermatozoal migration rather than by direct effect of copper on sperm motility.
The proteolytic fragments of the nuclear estrogen receptor in the MCF-7 cell line were characterized following limited digestion with chymotrypsin and trypsin. Nuclei were isolated from cells previously exposed to 10 nM [3H]estradiol. The proteolytic digestion was performed either on the micrococcal nuclease hydrolysate or on intact nuclei. The molecular weights (Mr) were calculated from the sedimentation coefficients determined on a sucrose gradient and from the Stokes radii estimated by gel filtration. Digestion of the nuclei with micrococcal nuclease solubilized a receptor form of Mr = 151,000. This receptor form was degraded by chymotrypsin to a receptor of Mr = 33,000 and by trypsin to a receptor of Mr = 60,000. Digestion of intact nuclei with chymotrypsin solubilized a receptor form of Mr = 62,000 which dissociated in 0.4 M KCl to a receptor of Mr = 32,000. Digestion of intact nuclei with trypsin followed by micrococcal nuclease solubilized a receptor form of Mr = 75,000 which was further dissociated by 0.4 M KCl to a receptor form of Mr = 60,000. The ability of the receptor forms to bind DNA was tested using DNA-cellulose column chromatography. About 40% of the micrococcal nuclease solubilized receptor form, compared to about 7% of the chymotrypsin degraded receptor and to about 13% of the trypsin degraded receptor forms, all bound to the column and could be eluted by high salt concentrated buffer. We conclude that the nuclear estrogen receptor in the MCF-7 cell line can be partially degraded either in the micrococcal nuclease hydrolysate or in intact nuclei by chymotrypsin or trypsin generating protein moieties, probably receptor fragments of Mr = 33,000 and 60,000 respectively. Both fragments retain their estradiol binding domain and it may be hypothesized that the heavier fragment retains its chromatin binding domain.
The assumption that a different conformational form was induced in the nuclear estrogen receptor following binding by antiestrogens compared to estrogens was studied by analysing the proteolytic fragments of the receptor following limited digestion with chymotrypsin and trypsin. Nuclei were isolated from MCF-7 cells previously exposed to [3H] 4-OHTAM. The proteolytic digestion was performed either on the micrococcal nuclease hydrolysate or on intact nuclei. The molecular weights (Mr) were calculated from the sedimentation coefficients (S) determined on a sucrose gradient and from the Stokes radii (Rs) estimated by gel filtration. Digestion of the nuclei with micrococcal nuclease solubilized a receptor form of Mr = 155,000. This receptor form was degraded by chymotrypsin to a receptor of Mr = 63,000 which could not be further dissociated by 0.4 M KCl and 3 M urea. A similar receptor molecule was released by chymotrypsin from intact nuclei. Digestion of the micrococcal nuclease hydrolysate with trypsin degraded the receptor to a form of a Mr = 67,000 which could not be further dissociated by 0.4 M KCl and 3 M urea. Digestion of intact nuclei with trypsin followed by micrococcal nuclease, solubilized a receptor form of Mr = 80,000 which could be further dissociated with 0.4 M KCl and 3 M urea to a receptor form of Mr = 67,000. This trypsin degraded receptor form seems to be similar in Mr to the chymotrypsin degraded form. On the other hand different receptor fragments of Mr = 33,000 and Mr = 60,000 were excised by chymotrypsin and trypsin respectively from the estradiol ligated estrogen receptor. (Geier et al., J. steroid Biochem. 26 [1987] 35-40.) These results support the assumption of a different conformational form for the antiestrogen ligated receptor, compared to the estrogen ligated receptor since they were differentially susceptible to proteolytic degradation by chymotrypsin.
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Progesterone (P) receptor levels were measured in the cytosol and in the 0.4 M KCl nuclear extract in human cervical tissues with [3H]-R5020 as a ligand and the results compared with those obtained in the myometrium and endometrium of the same uteri. Tissue samples were obtained from 28 women, grouped as follows: group A, 12 premenopausal controls; group B, 7 perimenopausal women who received estrogen 7 to 11 days before operation; and group C, 9 perimenopausal and postmenopausal women who received estrogen as in group B plus P injection 1 to 3 hours before operation. Estrogen administration resulted in a significant rise in total P receptor levels in the cervix, compared with the endometrium and myometrium. P injection after estrogen priming resulted in down-regulation of the P receptor in the cervix to undetectable levels, whereas in the endometrium and myometrium it resulted only in redistribution of the P receptor, with higher nuclear levels and lower cytosol levels. These results indicate the possibility of a different mechanism of regulation of the P receptor in the human cervix, compared with that in the endometrium and myometrium.
Monitoring of human menopausal gonadotropin (hMG) treatment for induction of ovulation according to either preovulatory estrogen levels or the presence of a dominant ovarian follicle was found insufficient to prevent ovarian hyperstimulation syndrome (OHS). In 65 infertile patients treated with hMG and human chorionic gonadotropin (hCG), a possible correlation between the number and size of all ovarian follicles on the day of assumed ovulation and the occurrence of OHS was evaluated in order to assess the value of ultrasonography in predicting OHS. It was found that patients with OHS had significantly more follicles at the time of hCG than patients without OHS. Mild OHS was characterized by the presence of eight to nine follicles, 68.7% of which were of intermediate size (9 to 15 mm). In moderate to severe OHS 95% of the preovulatory follicles were less than 16 mm, most of them (54.7%) less than 9 mm in diameter. It can be concluded that a specific preovulatory follicular configuration characterizes mild and severe hyperstimulation. This is important information before hCG administration and emphasizes the value of ovarian ultrasonography in predicting OHS.
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To explore further the relation between infertility and breast and female reproductive cancers, cancer incidence among 2,632 Israeli women treated for infertility between 1964 and 1974 was evaluated. Cancer incidence through December 1981 was determined by matching the study cohort to the Israel Cancer Registry. The observed number of cancers was compared with sex-age-ethnic and calendar-year, site-specific national cancer rates. There were 42 cancers observed compared with 37.4 expected, yielding a standardized incidence ratio of 1.1 (95% confidence interval (CI) = 0.8-1.5). Analysis by infertility diagnosis demonstrated no significant excess of total cancer incidence; the standardized incidence ratio was 1.3 (95% CI = 0.8-1.8) for infertility due to hormonal deficiency, 0.7 (95% CI = 0.3-1.4) for mechanical infertility, 1.6 (95% CI = 0.6-3.6) for infertility of the male partner, and 1.1 (95% CI = 0.5-2.2) for unclassified diagnosis. Site-specific analyses revealed a significantly increased risk (8.0; 95% CI = 2.5-19.3; four cases observed, 0.50 expected) of endometrial cancer for the hormonal group and a nonsignificant excess of breast cancer and melanoma. Although numbers were small, women with disorders causing unopposed estrogen production had a risk of 1.4 (95% CI = 0.8-2.2) for all cancer sites, which reached 10.3 (95% CI = 2.6-28.2; three cases observed, 0.29 expected) for endometrial cancer and 1.8 (95% CI = 0.8-3.4; eight cases observed, 4.43 expected) for breast cancer. Among women with nonhormonal infertility, there was a suggestion of increased risks of carcinoma of the ovary (3.2; 95% CI = 0.3-32.9; two cases observed, 0.63 expected) and thyroid (3.0; 95% CI = 0.3-24.6; two cases observed; 0.67 expected). No evidence of an association between ovulation-inducing drugs and cancer was found. This study supports the hypothesis that infertility caused by hormone deficiency is a risk factor for uterine cancer, but is inconclusive regarding breast cancer.
Nuclear progesterone receptors (RPN) were characterized in the 0.4 M KCl nuclear extract of the endometrium using [3H]R5020 as a ligand, in women in the proliferative phase of the cycle (n = 5), in the midcycle (n = 5), in the secretory phase (n = 4), and in premarin-treated women (n = 3), 1-3 h following progesterone injection. The receptor character of the binding was demonstrated by high specificity for progestin binding, high saturable affinity for R5020 and a sedimentation constant of about 3S in sucrose density gradient. The RPN characteristics were similar in all 4 groups and were also similar to those of the remaining cytosolic RP. Despite the fact that total RP levels differed in all 4 groups, the percentage of the receptor that was measured in the nuclear extract was similar in all groups and ranged from 52 to 61% of the total RP levels.
Human seminal plasma from normal or patients with abnormal parameters of the ejaculates contains an inhibitory material that expresses potent in vitro inhibitory activity on PHA-M-induced blast transformation and NK activity. Using the test of inhibition of NK activity, the semen samples from individuals with higher concentrations of fructose had higher inhibitory activity. The results described herein suggest that inhibitory activity for blast transformation may be present in the prostatic fluid while the NK inhibition aspects are correlated with the vesicle-marker (fructose). Inhibition of the immune responses by human seminal plasma of the effector functions indicates the interesting implication that soluble factors may indirectly protect against or promote human autoimmune infertility disease.
Progesterone receptor (RP) levels were measured in the cytosol and in the 0.4 M KCl nuclear extract of human uterine leiomyoma, myometrium and endometrium; the synthetic progestin [3H]-R5020 was used as a ligand. The women were divided into groups A-D as follows: A, 7 women with no treatment; B, 7 women who received progesterone injection 1-3 h before tissue collection; C, 4 women who received conjugated estrogens 7-11 days before tissue collection; D, 5 women who received both conjugated estrogen and progesterone treatments. The following results were demonstrated: Total RP levels in the leiomyoma were significantly higher than the corresponding levels in the myometrium of the same uterus; after estrogen treatment these differences diminished. Leiomyoma challenged with progesterone expressed significantly lower RP levels in their nuclei than did the corresponding endometrium and myometrium. These results indicate the possibility of a defect of RP dynamics in human uterine leiomyoma.
The effect of unilateral testicular torsion on the contralateral testis and the fertility rate was studied in Charles River adult rats. Animals were divided into groups that underwent a sham operation or torsion and ligation of the left testicular vessels followed by orchiectomy after 24 h, orchiectomy after 48 h, release of the ligature after 24 h, release of the ligature after 48 h, and no further treatment following ligation. Another group of animals underwent unilateral orchiectomy. After 8 weeks animals were allowed to mate and were sacrificed 2 weeks later. The results did not point to either histological alterations in the contralateral testis or impairment of fertility in any group of treatment compared with the control.