Effects of tamoxifen and estradiol on estrogen binding sites in the urogenital tract: an experimental study in the rabbit.

The aim of the study was to investigate the effects of estradiol and tamoxifen alone and in combination on the estrogen binding site status of the urogenital tract in the rabbit. Bilaterally ovariectomized rabbits were divided into four groups of six. Whereas the control group received no treatment, the remaining rabbits were treated with estrogen or/and tamoxifen. Cytosolic and nuclear fractions were isolated from the uterus, vagina, urethra and urinary bladder and used for binding site assay, by radioligand binding. The total weight of the rabbit vagina and uterus was increased significantly by both estradiol, tamoxifen and the combination of the two. The total weight of the urethra was increased only in the combination group. The cytosol binding site was downregulated by estradiol, tamoxifen and combination in the uterus, and in the vagina. Cytosol binding site in the urethra was not detected. The combination of estrogen-tamoxifen markedly reduced the nuclear binding site in the urethra and decreased affinity of the nuclear binding sites in all three tissues. The data suggest that tamoxifen has a specific ability to modulate the transcriptional activity of the estrogen binding sites in the rabbit urogenital tract.

Peripheral benzodiazepine receptor in human endometrium and endometrial carcinoma.

Since elevated concentrations of peripheral benzodiazepine (PBZr) have been reported in some tumors it was of interest to study the distribution PBZr in the endometrium and endometrial carcinoma. A specific ligand or PK 11195 was used to measure PBZr in the mitochondria (m-fraction) and the microsomal fraction (p-fraction). Density of binding sites and binding affinity (KD) was computed from Scatchard analysis. Binding in both m- and p-fractions from endometrium and endometrial carcinoma were saturable and of high affinity. The PBZr density in p-fraction from endometrium was slightly higher than in m-fraction, whereas the binding in m-fraction in endometrial carcinoma was more than 2-fold higher than in p-fraction. The difference between the densities in m-fraction from endometrial cancer and endometrium was highly significant (p < 0.005). There was no significant difference in the binding affinities (KD values) for PK 11195 in the various fractions. The selective increase in the density of PBZr in mitochondria of tumors suggests an association with energy demand and utilization and could be of diagnostic and/or therapeutic value in endometrial cancer. The role of a relatively high density of PBZr in microsomal fractions remains to be elucidated.

Elevated concentrations of mitochondrial peripheral benzodiazepine receptors in ovarian tumors.

Using PK 11195, a high affinity ligand for peripheral benzodiazepine receptor (PBZr) binding sites in isolated mitochondria (m-fraction) and microsomal fraction (p-fraction) from human ovaries and ovarian tumors were studied. Binding in both m and p-fractions from ovaries and tumors were saturable and of high affinity. The PBZr density in m- and p-fractions from ovaries was very similar whereas the binding in m-fraction in ovarian tumors was nearly 3-fold higher than p-fraction. There was no significant difference in the binding affinities (KD values) for PK 11195 in the various fractions. The selective increase in the density of PBZr in mitochondria of tumors suggests an association with energy demand and utilization. The role of a relatively high density of PBZr in microsomal fractions remains to be elucidated.

Identification of TP53 gene mutations in uterine corpus cancer with short follow-up.

The involvement of the TP53 tumor suppressor gene in uterine corpus cancer was investigated by single-stranded conformation polymorphism and sequence analysis of its exons 4 to 10. Mutations were found in 12 (18.5%) of 65 cases. Ten of these 12 were single-base substitutions (8 missense and 2 nonsense mutations), whereas 2 were frame-shifting mutations. TP53 gene mutations correlated significantly with advanced surgical stage of disease (P = 0.006) and unfavorable tumor histology types (P = 0.003), whereas the association to myometrial wall invasion did not reach statistical significance (P = 0.054). TP53 gene mutations also correlated significantly with allelic loss at TP53 locus (P = 0.024), absence of estrogen (P = 0.045) and progesterone receptors (P = 0.001), DNA nondiploidy (P = 0.002), and high S-phase fraction values (P = 0.002). Our results suggest that inactivation of the TP53 checkpoint function is associated with disease transition into a stage of rapid progression and spread.

Karyotypic characteristics of borderline malignant tumors of the ovary: trisomy 12, trisomy 7, and r(1) as nonrandom features.

Clonal karyotypic abnormalities were detected in five of 14 cytogenetically analyzed borderline malignant ovarian tumors of clinical stages I-II. One mucinous and one seropapillary tumor had trisomy 7 and r(1)(p36q42) as the sole chromosome abnormality, respectively. Trisomy 12 was found in the remaining three cases. It was the only change in one mucinous and one serous tumor, whereas the third, a seropapillary borderline tumor, had the karyotype 49,XX,+5,+8, +12. These findings, especially when collated with those of previous reports on ovarian borderline tumor cytogenetics, indicate that +12 is the most consistent chromosomal aberration in this group of neoplasms and that also +7 and r(1) are nonrandom features. From the karyotypic point of view, benign ovarian tumors and well-differentiated carcinomas are similar to borderline ovarian tumors, with the possible exception that the former have no tendency to form r(1). Highly malignant carcinomas, on the other hand, are typically much more complex. Chromosome-level changes therefore cannot account for the putative phenotypic passage through the most innocuous tumor stages as epithelial ovarian neoplasms go from benign to fully malignant.

Elevated concentrations of antiestrogen binding sites in membrane fractions of human ovarian tumors.

The density and affinity of tamoxifen (TAM) binding sites in isolated plasma membrane fractions from human ovarian tissue and ovarian tumors was measured. TAM binding in both membrane preparations was specific, saturable, and had a high affinity (KD < 1 nM). The density of TAM binding sites in the tumors was at least three times higher than that in the normal ovaries, whereas the affinity of TAM binding sites in the tumors was lower than in normal ovaries. The relatively large number of TAM binding sites that are distinct from estrogen receptors in ovarian tumors suggest that TAM alone and in combination with other chemotherapy could be used with advantage in ovarian cancer.

Laparoscopic surgery for stress urinary incontinence.

Retropubic bladder neck suspension can easily be performed laparoscopically giving the same therapeutic result as open surgery. Laparoscopy enables better visibility of the operative field, exact placement of paraurethral sutures and acceptable hemostasis. Patients who undergo laparoscopic retropubic suspension meet with less postoperative discomfort, fewer operative complications, a shorter hospital stay and less postoperative scars. The patients are allowed to drive and return to work 1-2 weeks after surgery.

Laser "miniconisation' and the outcome of subsequent pregnancy.

We routinely use the carbon dioxide laser miniconisation procedure for cervical interepithelial neoplasia I-III. A retrospective case control study was conducted in 1,485 miniconised women during 250 subsequent pregnancies (including three twin pregnancies). The analysis showed that the miniconised subjects had no significant pregnancy complications and the same preterm birth ratio (8.0%) as controls matched for age and parity.

Insidious urinary retention after vaginal delivery: prevalence and symptoms at follow-up in a population-based study.

The purpose of this study was to investigate the prevalence of postpartum urinary retention in women after vaginal delivery and to determine whether parturients with retention develop voiding problems later. During a 3-month period, all parturients in the catchment area of the University Hospital, Lund, were investigated 3 days after delivery, residual volume being measured by ultrasonography. All those with postpartal retention were contacted 4 years after delivery, when they were reexamined by ultrasonography and asked to fill in a questionnaire regarding urinary problems. In all, 539 women were scanned post partum, and 8 (1.5%) had a residual volume exceeding 150 ml (range 156-320 ml). Retention was more common among primiparae after instrumental delivery or epidural analgesia. The symptoms were normalized spontaneously within a few days in all cases. At follow-up 4 years later, the prevalence of urinary symptoms was not higher than that in the general population. Ultrasonography to detect urinary retention does not seem to have any place in the normal postpartal care. However, extended supervision may be appropriate in parturients receiving epidural analgesia or in those submitted to instrumental deliveries.

Effects of long-term treatment with estrogen and progesterone on in vitro muscle responses of the female rabbit urinary bladder and urethra to autonomic drugs and nerve stimulation.

Ovariectomized virginal rabbits were treated with either estrogen or progesterone for 4 to 6 months. In vitro responses of muscle strips of the bladder and circular urethra were examined. Estrogen resulted in bladder contractions in response to noradrenaline and phenylephrine, whereas these agonists were without effect or evoked weak relaxations in castrated animals (and normals). Estrogen also caused a rightward shift of the frequency-contraction curve to nerve stimulation. Progesterone increased bladder sensitivity to contraction-evoking bethanechol. Contractile urethral responsiveness to bethanechol increased after both steroids. Urethral sensitivity to noradrenaline, evoking contraction, increased following estrogen. Further, estrogen abolished the marked relaxatory urethral response to nerve stimulation of castrated (and normal) rabbits and caused contraction only, which was abolished by a combination of alpha-adrenoceptor antagonists and scopolamine. When comparing the present results with those of other studies, it is evident that hormone-induced changes become manifest at an early stage. The present animal study gives support for the use of estrogen in the therapy of stress incontinence and, further, it provides no objections to the use of progesterone in combination with estrogen in this condition.

Surgical results and urodynamic studies 10 years after retropubic colpourethrocystopexy.

Retropubic colpourethrocystopexy has been shown to be associated with good immediate cure rate in treating female urinary stress incontinence. Urodynamic data is often lacking in follow-up studies. This study presents long-term results 8-12 years after operation. Between 1979-1982, 72 women suffering from genuine stress incontinence underwent a colpourethrocystopexy. Follow-up was done 1989-1990. Each patient had a complete urodynamic evaluation preoperatively and at the follow-up. Mean age at operation was 46.2 years and follow-up was done on average 9.4 years later (8-12 years). 71% were postmenopausal at the time of follow-up. Surgical cure rate was 90.3%, 9.7% (seven patients) were considered failures. Five of the seven patients had experienced much improvement. A number of patients showed a decrease in one or more urodynamic parameter without a recurrence of stress incontinence. The bladder neck was incompetent in 38 (52.8%) of the patients, 31 of the 38 patients were still continent. Retropubic colpourethrocystopexy should be considered the primary operation for the treatment of female stress incontinence. Cure rate and postoperative evaluation cannot be judged by urodynamic parameters only, as some of the continent women showed a decrease in one or more urodynamic parameters that are considered important for continence. Bladder neck competence is not necessary for postoperative continence.

Abdominal sacral colpopexy with use of synthetic mesh.

Forty patients with post-hysterectomy vaginal vault prolapse underwent surgery at Lund University Hospital between 1983 and 1991. The standard procedure was an abdominal colposacropexy, using Marlex mesh in 25 cases and Gore-Tex mesh in 15. There were no intraoperative complications. One patient developed a recurrent vault prolapse and one patient developed a draining sinus in the vaginal apex. The duration of follow-up has varied between one and ten years. In 39 of the 40 patients (97%), both subjective and objective improvement has been achieved.

Characterisation of muscarinic cholinergic receptors in human ovaries, ovarian tumours and tumour cell lines.

Using [3H]quinuclidinyl benzilate (QNB) as radioligand, muscarinic cholinergic receptor sites in isolated plasma membrane fractions from human ovarian tumours, cultured tumour cells, and normal ovarian tissue were characterised. QNB binding to all preparations, except from poorly differentiated tumour, was specific, saturable, and of high affinity. In contrast to normal ovaries, benign tumours, well differentiated adenocarcinoma and OVCAR-3 cells, the poorly differentiated adenocarcinoma and SKOV-3 cells completely lacked specific QNB binding. The muscarinic receptor densities and the Kd values in preparation from ovaries, receptor-positive tumours and OVCAR-3 cells were similar. QNB binding was strongly inhibited by the classical muscarinic receptor antagonist atropine, but poorly by the agonist carbachol. In contrast to atropine, inhibition by pirenzepine and AF-DX 116 was relatively low. These data suggest that muscarinic receptors in ovaries and ovarian tumours are of m3 type.

Effect of estrogen treatment on the peroxidase activity and estrogen receptors in the female rabbit urogenital tissues.

The effect of one, 4 and 8 weeks of continuous estrogenization of rabbit on peroxidase (PO) activity and on both cytoplasmic and nuclear estrogen receptors was studied in the uterus, vagina, urethra and urinary bladder. Whereas the peroxidase activity in the urogenital tissue of untreated controls was near zero, after one week of estrogen treatment it increased very substantially in the uterus and vagina and much less dramatically in the urethra or urinary bladder. With continuation of estrogen treatment for 4 or 8 weeks the PO decreased by 80-90% in the uterus and vagina and by only 40% in the urethra. After one week of estrogen treatment the density of both cytosolic and nuclear estrogen receptors decreased by several fold in both the uterus and vagina, whereas it decreased by 30-50% in the urethra and bladder. The concentration of both cytosolic and nuclear receptors decreased further although less dramatically with continuing estrogen treatment, up to 8 weeks, in all tissues. These data suggest that although the general pattern of responses to estrogen showing an initial increase in PO followed by a reduction with continuing estrogen treatment is the same in all urogenital tissues, the responses seem to have a prolonged time scale in the case of lower urinary tract tissues. The quantitative aspect of the response generally corresponds with the density of estrogen receptors in the urogenital tissues.

Effects of protracted administration of estriol on the lower genito urinary tract in postmenopausal women.

We investigated 80 postmenopausal women, 48 of whom (60%) agreed to undergo long-term treatment with estriol suppositories. All had symptoms of vaginal atrophy and urinary incontinence. Endometrial samples were taken after 8-10 years of therapy. Estriol had induced slight proliferative changes in the endometrium in 7 of 48 patients studied by endometrial sampling. 75% of the women reported significant subjective improvement of stress incontinence. Estriol supplementation did not produce any significant change in urethral pressure, functional length, or cystometric parameters. However, a significant increase in pressure transmission ratio to the proximal urethra was noted after vaginal medication with estriol. Replacement therapy in post-menopausal women must take into account the patients perception of risks and benefits. The risk of estriol treatment is insignificant.

Tissue specific effects of progesterone on progesterone and estrogen receptors in the female urogenital tract.

The effect of progesterone administration on progesterone and estrogen receptors in the uterus, vagina and urethra of rabbits was studied. After 24 h of progesterone treatment the concentration of cytosolic progesterone receptors decreased to about 25% of the control value in the uterus, whereas no significant change in receptor concentration was observed in the vagina or the urethra. The concentration of the nuclear progesterone receptor did not change in any of the three tissues studied. The apparent dissociation constant (Kd) of nuclear progesterone receptor increased after progesterone treatment in all three tissues. Although the Kd of the cytosolic progesterone receptor also increased in all tissues, the difference was significant for only the vagina and urethra. The concentration of cytosolic estrogen receptors in the uterus decreased significantly (P less than 0.001) after progesterone treatment whereas the Kd value increased slightly (P less than 0.05). In vagina or the urethra, there was no change in either estrogen receptor concentration or Kd values after progesterone treatment. These data clearly showed that the reduction by progesterone of progesterone and estrogen receptor concentrations occurs only in the uterus and not in the vagina or the urethra.

Prevalence of urinary incontinence in middle-aged women.

An anonymous questionnaire was used to interview 946 premenopausal women (age 29-52 years), previously either hysterectomized or laparoscopically sterilized, concerning disorders of the urinary tract. Of the 750 (79.3%) women that responded, 307 (40.9%) admitted to some degree of urinary disorder. The prevalence of urinary disorders was 170/415 (40.9%) for the women with a previous supravaginal or total hysterectomy and 137/335 (40.8%) for the women in the sterilization group. Of the responding women 85 (11.3%) experienced a urinary loss sufficient to necessitate the wearing of a sanitary napkin or change of underclothing several times a day.

Urodynamic studies before and after retropubic colpo-urethrocystopexy in fertile women with stress urinary incontinence.

We compared urodynamic parameters in a group of 30 healthy women of fertile age, before and after primary retropubic colpo-urethrocystopexy carried out to correct genuine stress urinary incontinence. The mean interval between surgical treatment and follow-up was 38.0 (SD 10.2) months. In 26 women, subjectively and objectively cured at follow-up, we found an increase of 5.1 mm in functional length of the urethra (t = 4.5; P less than 0.001), an increase of 12.3 cm H2O in the urethral closure pressure (t = 3.9; P less than 0.001), and a 22.1% increase in the transmission ratio (t = 3.2; P less than 0.01). However, 9 of these 26 women satisfied with the results of the surgical treatment showed a drop in at least 2 of the parameters studied. In the group of 4 women not satisfied with the results of the surgical treatment we found an improvement in all of the urodynamic parameters studied, similar to those found in the group of satisfied women, though less pronounced. This study indicates that successful surgical treatment of genuine stress urinary incontinence is not related to surgically induced changes in the urodynamic parameters.

Sexual function after colpo-urethrocystopexy in middle-aged women.

This is the first prospective study to analyse sexual function in women following an incontinence operation. The author has personally interviewed and examined 156 patients, first before and then 1-2 years after operation. 32% of the women with stress incontinence had had sexual problems before surgery. This proportion decreased after surgery to 10%. Colpo-urethrocystopexy elevates the anterior vaginal wall and tilts the axis of the vagina. These corrections do not affect the sexual function of the vagina. None of the patients developed dyspareunia after surgery.

Progesterone receptors in the female lower urinary tract.

When female estrogenized rabbits were injected i.v. with 3H-progesterone, the tritium concentration determined after one hour was about two to three times higher in urethra, urinary bladder and vagina than in the heart. High affinity progesterone receptors (KD = 1-2 nM) could be demonstrated in both cytoplasmic and nuclear fractions prepared from estrogenized rabbit urethra, bladder and vagina. The cytosolic receptor concentration in both urethra and bladder was about half of that in the vagina. The concentration of nuclear receptors in urethra was not significantly different from that in the vagina, but in the bladder the concentration was only about one fourth of that in the vagina or urethra. The mean KD of cytosolic receptors from bladder was significantly higher than the corresponding values in urethra and vagina. Progesterone binding sites in the bladder had a broader hormonal specificity than those in the urethra or vagina. The present demonstration of specific progesterone receptors in the female urethra might provide a possible link between estrogen progesterone interaction and the appearance of urinary incontinence during pregnancy in women.