Differential expression of p73 splice variants and protein in benign and malignant ovarian tumours.

The p73 gene encodes a protein with substantial structural and functional similarities to the tumour-suppressor p53. Alternative splicing of p73 mRNA leads to expression of 6 known RNA species and proteins (alpha, beta, gamma, delta, epsilon, zeta). We analysed the expression of these splice variants in ovarian adenocarcinoma by RT-PCR followed by detection of amplicons with the Southern technique and by immunoblot in 32 malignant and benign epithelial ovarian tumour specimens and 3 ovarian adenocarcinoma cell lines (A2780, 2008, OVCAR-3). p73alpha mRNA was expressed in all 17 ovarian cancer specimens, and 14 of 17 expressed at least 3 splice variants. In contrast, a different expression pattern was present in the ovarian adenomas: p73alpha was detected in 6 of 12 benign tumours, and only 1 adenoma expressed 3 splice variants. p73 protein was expressed in 9 of 16 ovarian cancer specimens, in all cell lines and in 1 of 3 borderline tumours. In contrast, none of 9 ovarian adenomas expressed detectable amounts of p73 protein. Expression of p73 mRNA and protein was not correlated with FIGO stage and histological grade, but we observed a significant correlation with over-expression of p53 protein. In summary, epithelial ovarian cancers express a more complex p73 isoform pattern and higher levels of p73 mRNA and protein than ovarian adenomas.

Decrease of serum endothelin levels with postmenopausal hormone replacement therapy or tibolone.

Endothelin is the most potent vasoconstrictor peptide known to date. Hormone replacement therapy (HRT) with estrogen reduces plasma endothelin levels. We measured endothelin in 51 postmenopausal patients before and during HRT. Patients were randomly allocated to receive either oral tibolone, oral or transdermal 17 beta-estradiol. A group of comparable volunteers served as controls. After 24 months, endothelin levels decreased in all treatment groups: tibolone, 18.2%; oral 23.1%; transdermal, 20.8%. Endothelin levels increased in the controls by 36.6% (p < 0.01). Tibolone decreases endothelin levels to a similar degree as conventional estrogen-progestogen-replacement therapy. These data provide another potential mechanism supporting the cardioprotective effects of tibolone.

The long-term tolerability and efficacy of OESCLIM: results of a 1-year study.

OBJECTIVES: A 1-year, open-label, non-comparative study evaluated the long-term tolerability and acceptability of a new generation matrix patch in post menopausal women with estrogen deficiency. METHODS: Menopausal women (224) from 37 centres in five European countries received OESCLIM 50 microg/d (17-beta estradiol) for 3 months, titrated if necessary to either 25 or 100 microg/d for a further 9 months. Patients received either a continuous or discontinuous estradiol regimen with concomitant sequential progestogen (except hysterectomised patients). Skin tolerability was assessed by patient diaries and questionnaires. Global tolerability, efficacy, laboratory parameters and global acceptability were also monitored. RESULTS: Almost two-thirds of women did not experience any kind of skin reaction and only 4.3% of all applications (752/17,702) caused site reactions. Of these, the majority caused only slight or no discomfort (63.2%). Only 0.37% of total applications required patch removal; none required therapy. A low percentage of patients withdrew due to tolerability issues: 2.7% due to skin reactions; 7.5% due to hyperestrogenism. The mean number of hot flushes experienced by symptomatic women reduced by 91% from 4.0 at baseline to 0.4 after 2 months. Total cholesterol reduced by 3.9% and LDL cholesterol by 5.1%, with no increase in triglyceride levels. Investigators assessed treatment as effective in 96.8% of cases; well tolerated locally in 93.1% and well tolerated generally in 89.5%. At the end of this 1 year study, 79% of patients wished to continue therapy. CONCLUSION: OESCLIM is well tolerated locally and systemically in long-term therapy with a high proportion of patients wishing to continue therapy after 1 year.

Prevention of postmenopausal bone loss using tibolone or conventional peroral or transdermal hormone replacement therapy with 17beta-estradiol and dydrogesterone.

Postmenopausal bone loss can be prevented by continuous or intermittent estradiol (E2) administration. Concomitant progestogen therapy is mandatory in nonhysterectomized women to curtail the risk of endometrial hyperplasia or cancer. However, the recurrence of vaginal bleeding induced by sequential progestogen therapy in addition to continuous estrogen administration is one of the reasons for noncompliance to hormone replacement therapy (HRT). Tibolone, a synthetic steroid with simultaneous weak estrogenic, androgenic, and progestational activity, which does not stimulate endometrial proliferation, has recently been proposed for the treatment of climacteric symptoms. To compare the efficacy of conventional oral and transdermal HRT with that of tibolone in the prevention of postmenopausal bone loss, 140 postmenopausal women (age, 52 +/- 0.6 years; median duration of menopause, 3 years) were enrolled in an open 2-year study. Volunteers had been offered a choice between HRT and no therapy (control group, CO). Patients selecting HRT were randomly allocated to one of the following three treatment groups: TIB, tibolone, 2.5 mg/day continuously, orally; PO, peroral E2, 2 mg/day continuously, plus sequential oral dydrogesterone (DYD), 10 mg/day, for 14 days of a 28-day cycle; TTS, transdermal E2 by patch releasing 50 microg/day, plus DYD as above. Bone densitometry of the lumbar spine, upper femur, and whole body was performed using dual-energy X-ray absorptiometry at baseline, and then 6, 12, 18, and 24 months after initiation of therapy. One hundred and fifteen women (82%) completed the 2 years of the study. The dropout rate was similar in each group. Over 2 years, bone preservation was observed in all three treatment groups as compared with controls, without significant differences among treatment regimens. In conclusion, tibolone can be regarded as an alternative to conventional HRT to prevent postmenopausal bone loss.

Placental protein 14 (PP14) does not predict endometrial status under hormone replacement therapy.

Our objective was to compare serum level of placental protein 14 (PP14) with histological findings in endometrial evaluation of postmenopausal women using hormone replacement therapy (HRT). In a subset of 109 out of 140 women included in a randomized comparative study, serum levels of PP14 were determined after 12 months of use of (1) no HRT; (2) oral micronized 17 beta-estradiol/oral sequential dydrogesterone; (3) transdermal 17 beta-estradiol/oral sequential dydrogesterone; or (4) oral tibolone. Subjects underwent Pipelle biopsy after 12 months. The serum level of PP14 was determined by sandwich enzyme immunoassay (ELISA). The two-tailed t-test and one-way ANOVA or their non-parametric equivalents were used to test for statistical significance. All three HRT regimens were safe with respect to the endometrium. Hyperplastic or malignant changes were not observed. There was a significant difference in the mean values of PP14 between the groups of inactive/atrophic and secretory endometrium (p < 0.01). However, there was a wide range of individual values for PP14 within the groups and a wide overlap in values between the groups of non- substituted and hysterectomized women. The quantitative determination of PP14 in the serum did not provide supplementary information on the substituted endometrium. From this study it can be concluded that the serum PP14 determination is not useful to predict endometrial status under HRT. The relatively high levels of PP14 in hysterectomized patients suggest ectopic production.

Microscopic findings of the nail-fold capillaries--dependence on menopausal status and hormone replacement therapy.

The aim of our controlled study was to evaluate peripheral microcirculation at the level of the nail-fold capillaries in relation to menopause status and postmenopausal hormone replacement therapy (HRT). A total of 105 postmenopausal women were randomly allocated to three different HRT groups of equal size. A fourth group of 35 similar healthy volunteers served as controls. HRT was either peroral or transdermal 17-beta-oestradiol with cyclic addition of dydrogesterone or 2.5 mg Tibolone (Org OD 14) in a daily peroral dose. Morphological parameters such as capillary diameters, loop width, papillary width and capillary density, measured by video-capillaroscopy at the nail-fold, were unaffected in early menopause and also under HRT. A significant decrease of capillary blood flow velocity (P < 0.001) could be demonstrated in postmenopausal (n = 41, v = 0.53 +/- 0.16 mm/s) as compared to premenopausal women (n = 37, v = 0.65 +/- 0.15 mm/s). HRT resulted in an increase of capillary blood flow velocity in the nail-fold after 6 and 12 months leading to an increase in capillary blood flow in the order of 20%-30% of the initial values, and was independent of the type of HRT.

Curative treatment of clinical stage II endometrial carcinoma.

In a retrospective study of 54 patients we compared three treatment policies of clinical stage II endometrial carcinoma: One group of patients (n = 29) was surgically staged with histological examination made at surgery and treated by abdominal total hysterectomy with bilateral salpingo-oophorectomy. In case of cervical infiltration by tumour, a pelvic node sampling was added. The second group was treated by radical hysterectomy (n = 9) and the third group of patients (n = 16) was treated by abdominal total hysterectomy with bilateral oophorectomy only. The median follow-up time was 68 months. The overall five year-survival was 64%. The overall five year-survival rates according to surgical procedure were not significantly different. The survival curves were significantly different from each other as were those according to surgical stage and age. In a multivariant analysis stage, grading and age proved to have an independent relation with the survival rate whereas type of surgical procedure, depth of myometrial infiltration and type of radiotherapy had no influence on survival. Intraoperative histological staging allows adaption of treatment to the true stage, thus minimising treatment-related morbidity.

Long-term results after Burch colposuspension.

OBJECTIVE: Our purpose was to review the long-term (5 to 10 years) clinical and urodynamic outcome in patients with stress urinary incontinence after Burch colposuspension. STUDY DESIGN: A follow-up of 87 women with stress urinary incontinence who had a Burch colposuspension between 1979 and 1985 at the Department of Obstetrics and Gynecology, University of Berne, was performed by clinical and urodynamic reevaluation of the patients. RESULTS: Stress incontinence was cured in 81.6% of patients. The cure rate was not significantly related to age, hormonal status, body weight, or previous surgical procedures for incontinence. Burch colposuspension stabilized the urethrovesical junction. Urodynamic measurement at follow-up compared with the preoperative evaluation showed in the cured group a significant increase in (1) the functional urethral length at rest and at stress, (2) maximum urethral closure pressure at stress, and (3) pressure transmission. On the contrary, in unsuccessful operations none of the recorded parameters had improved. Women with failed surgery had significantly lower preoperative maximum urethral closure pressures at rest and at stress, lower continence areas, smaller functional urethral lengths at stress, smaller length to peak pressures, and lower index values of urethral relaxation at stress. The procedure had a low operative and postoperative morbidity, with no significant disturbance of voiding function noted at 5 to 10 years' follow-up. CONCLUSIONS: Our results with the Burch colposuspension showed a high success rate at 5 to 10 years' follow-up. The high cure rate and low operative and postoperative morbidity were related to careful preoperative selection.

Bone mineral density in young women with long-standing amenorrhea: limited effect of hormone replacement therapy with ethinylestradiol and desogestrel.

To assess bone mineral density (BMD) at different skeletal sites in women with hypothalamic or ovarian amenorrhea and the effect of estrogen-gestagen substitution on BMD we compared BMD of 21 amenorrheic patients with hypothalamic or ovarian amenorrhea with that of a control population of 123 healthy women. All amenorrheic patients were recruited from the outpatient clinic of the Division of Gynecological Endocrinology at the University of Berne, a public University Hospital. One hundred and twenty-three healthy, regularly menstruating women recruited in the Berne area served as a control group. BMD was measured using dual-energy X-ray absorptiometry (DXA). At each site where it was measured, mean BMD was lower in the amenorrheic group than in the control group. Compared with the control group, average BMD in the amenorrheic group was 85% at lumbar spine (p < 0.0001), 92% at femoral neck (p < 0.02), 90% at Ward's triangle (p < 0.03), 92% at tibial diaphysis (p < 0.0001) and 92% at tibial epiphysis (p < 0.03). Fifteen amenorrheic women received estrogen-gestagen replacement therapy (0.03 mg ethinylestradiol and 0.15 mg desogestrel daily for 21 days per month), bone densitometry being repeated within 12-24 months. An annual increase in BMD of 0.2% to 2.9% was noted at all measured sites, the level of significance being reached at the lumbar spine (p < 0.0012) and Ward's triangle (p < 0.033). In conclusion BMD is lower in amenorrheic young women than in a population of normally menstruating, age-matched women in both mainly trabecular (lumbar spine, Ward's triangle, tibial epiphysis) and mainly cortical bone (femoral neck, tibial diaphysis).(ABSTRACT TRUNCATED AT 250 WORDS)

Kallmann syndrome and associated malformation of the uterus.

Kallmann syndrome is a rare combination of hypogonadotropic hypogonadism due to hypothalamic insufficiency and anosmia. In both patients treated at our institution for infertility, a malformation of the uterus was noted: one patient had a unicornuate uterus, the other a uterus with a fundal hypoplasia and tubes of approximately 9 cm. It is not clear if the malformation is in association with Kallmann syndrome or purely coincidental. Given the known association of other malformations with Kallmann syndrome, we suppose that probably an underlying genetic defect having to do with organogenesis is the cause of the uterine malformations.

Benefits of different routes of administration.

The benefits of hormone replacement therapy (HRT) are well established for the different systemic administration routes used today. Relief from somatic and vasomotor postmenopausal symptoms can be obtained by all recognized forms of HRT. The prophylactic action of peroral and percutaneous administration of HRT on postmenopausal bone loss is identical if the appropriate protective dosage of the estrogen component is chosen for each galenic form. The second metabolic benefit of HRT, the cardioprotective effect of ERT, is well accepted for the peroral route. New data on the direct effect of estradiol on the arterial wall as well as recent serum lipid results obtained during percutaneous administration of estradiol in postmenopausal women allow the conclusion that the percutaneous route induces cardioprotection similar to that obtained with the classical peroral route. This opinion is supported by animal data. Furthermore, observations in animals suggest that the benefits of ERT are not neutralized by either sequential or fixed addition of progestin to the estrogen. Except in some rare cases with a particular indication for either the peroral or the percutaneous administration of HRT, the choice of route of administration should be made on the basis of maximum acceptability by the patient.

Postmenopausal hormone replacement therapy with Tibolone decreases serum lipoprotein(a).

Lipoprotein(a) is a cholesterol-rich plasma lipoprotein consisting of LDL and apolipoprotein(a). Apolipoprotein(a) shows structural similarity with plasminogen and thus may interfere with thrombogenesis. Lipoprotein(a) has been shown to be a strong independent risk factor for coronary heart disease. So far no drug or diet is known to have prominent effects on the serum levels of lipoprotein(a). In the present study we found a highly significant decrease (in the order of 26%) of lipoprotein(a) in 28 women treated for 6 months with Tibolone, compared with an age-matched healthy control group. Tibolone is a synthetic steroid with gestagenic and weak androgenic and oestrogenic properties, which shows no stimulation of the endometrium. Tibolone also produced a decrease in HDL-cholesterol of 23% (p < 0.001), a decrease in apolipoprotein A-I of 14% (p < 0.001) and an increase in apolipoprotein B of 17% (p < 0.001), whereas the control group showed no significant changes in these quantities. Tibolone in a daily dose of 2.5 mg is at present the only complete postmenopausal hormone replacement therapy that shows a significant inhibiting influence on serum levels of lipoprotein(a). Its effect on lipoprotein(a) might counterbalance, at least to some extent, the theoretical adverse effect on the other lipoprotein risk factors.

A new peroral estrogen/progestin combination for postmenopausal hormonal substitution: an open multicentric field study.

An open prospective multicentric trial has been conducted over 6 months in 241 postmenopausal volunteers. One-hundred forty-one women had an intact uterus. All patients received a fixed peroral combination of conjugated estrogens CE (1.25 mg per day from day 1 to day 21) and medrogestone (5 mg per day from day 12 to day 21) followed by 7 days without substitution (day 22 to day 28). After 3 months of treatment, the managing physician could, according to the patient's clinical response, reduce the dosage of CE to 0.625 mg daily. This dose reduction took place in 79 patients (38.9%). The trial was designed to study efficacy, compliance and side-effects of this combination. Of the patients 68.9% showed a very good, 27.7% a good and 1.9% a satisfactory improvement of their preexisting subjective complaints. Of the patients 28.6% suffered from minor side-effects leading to drop-outs in 7.8% of the cases. Of the women participating in the study 92.2% completed the trial without from the treatment scheme. No serious complications have been noted. After 6 months of treatment, a regular bleeding pattern has been observed in 71.5% of the 144 non-hysterectomized women, an irregular pattern in 9.7% and amenorrhoea in 18.8%. Total cholesterol showed no change, whereas HDL rose significantly from 1.58 to 1.72 mmol/l (P < 0.01) resulting in a drop of Total-Cholesterol-HDL-Ratio of -8.8% (P < 0.01). LDL decreased from 3.71 +/- 1.56 to 3.45 +/- 1.39 (P < 0.05). Considering the two patient groups with and without estrogen reduction after 3 months, HDL increase was significant in both groups but was dose dependent. The HDL increase compared to the initial value was 5.7% with 0.625 mg CE and +10.8% with 1.25 mg CE, respectively. The fixed peroral combination of CE and medrogestone tested was effective, easy to administer and safe. The bleeding pattern observed was mostly regular. The pattern of serum lipids changed favorably in a significant way. Therefore, the use of this new peroral estrogen/progestin combination can be recommended for routine substitution in postmenopausal women.

Combined bilateral tubal and multiple intrauterine pregnancy after ovulation induction.

Over the last 20 years, the frequency of multiple pregnancy has increased mainly because of the introduction of exogenous pituitary gonadotropins in the treatment of sterility. The incidence of ectopic pregnancies also increased during the same period of time. This paper describes the first reported case of a simultaneous bilateral tubal and multiple intra-uterine pregnancy after ovulation induction with human gonadotropins.

Urethral isolation of Chlamydia trachomatis in women with urinary incontinence.

The overall prevalence of Chlamydia trachomatis (CT) infection in 168 patients with urinary incontinence was 7.7%. However, in patients with urethral instability CT was found significantly more often than in those with stable urethra (P less than 0.001). Clinical and urodynamic improvement followed the treatment with doxycycline in 8 out of 9 patients with urethral instability. According to our results urethral infection with CT may play an important role in the etiology of urge incontinence and urethral instability.