Image analysis measurements of the microvascularisation in endometrium, superficial and deep endometriotic tissues.

UNLABELLED: The aim of this study was to evaluate precisely the microvascularisation of endometrium, superficial and deep endometriotic lesions, in progestin-treated and non-treated patients suffering from endometriosis. METHODS: A population of 66 women was constituted. Immunohistochemistry was carried out with a specific marker of the endothelial cells (CD31). The number of vessels and the vessel area were assessed by a computer image analysis system. RESULTS: The number of vessels per mm2 were 211, 216, 225 and the vessel area was 270, 141 and 194 microm2, respectively in endometria, superficial and deep endometriotic lesions of untreated women. In endometria, superficial and deep endometriotic lesions of progestin-treated women the number of vessels were respectively 129, 149, and 181 per mm2 and the vessel area was 369, 474 and 254 microm2. CONCLUSION: Statistically significant data indicate that endometriotic lesions are heterogeneous and suggest that progestin treatment induces a reduction in number and a concomitant dilation of microvessels with more microvascular changes in endometrium and superficial endometriotic lesions than in deep endometriotic lesions.

Impact of different progestins on endometrial vascularisation of postmenopausal women. A retrospective image analysis-morphometric study.

OBJECTIVES: To determine if the vascularisation of the endometrium is dependent on the administered progestin during sequential hormone replacement therapy. METHODS: Nine women received percutaneous estradiol-17 beta, 1.5 mg/day from days 1 to 24 combined with 200 mg/day micronised progesterone from days 11 to 24 of the treatment cycle. Fifteen women received percutaneous estradiol, 1.5 mg/day from days 1 to 24, combined with 10 mg/day chlormadinone acetate from days 11 to 24. Eleven women received percutaneous estradiol, 50 microg/day from days 1 to 28 combined with percutaneous norethisterone acetate, 0.3mg/day form days 14 to 28. Twelve women received intranasal estradiol, 300 microg/day from days 1 to 25 combined with 0.5 mg of promegestone from days 11 to 24. Eleven spontaneous cycling women had an endometrial biopsy during luteal phase and served as controls. Endometrial biopsies were processed routinely between days 18 and 24 and sections were immunostained using anti-CD34 antibody to identify vascular endothelial cells, which were treated with an automatic image analysis system. RESULTS: mean (+/-S.D.) vascular density for controls was 147+/-41.5 vessels/mm(2), with mean vessel area of 143+/-60.9 microm(2). In chlormadinone users endometrial microvascular density and mean vessel area did not differ from the control group (150.2+/-58.6 and 152.9+/-70.5). The other three progestins generated a significant increase of mean vessel density, 179.6+/-51.6 with micronised progesterone, 178.5+/-67.6 with norethisterone and 179.6+/-48.4 with promegestone. The mean vessel area was lower in the latter three groups, respectively, 108.4+/-39.0, 97.5+/-46.5 and 141.6+/-66.7 microm(2), promegestone leading to non significant difference with control. CONCLUSION: regarding vascularisation, chlormadinone and control group gave similar patterns. Promegestone was associated with an increase of the number of vessels, as did micronised progesterone or norethisterone; the mean vascular area was the smallest in the norethisterone group.

Comparative endometrial histology in postmenopausal women with sequential hormone replacement therapy of estradiol and, either chlormadinone acetate or micronized progesterone.

OBJECTIVES: To determine the endometrial response in postmenopausal women treated with a sequential hormone replacement therapy (HRT) of estradiol and, either chlormadinone acetate (CA) or micronized progesterone (MP). METHODS: Three hundred and thirty-six postmenopausal women with a normal endometrium were randomized in the double-blind study. All patients received percutaneous estradiol 1.5 mg/day from day 1 to day 24 and either CA 10 mg/day or oral MP 200 mg/day from day 10 to day 24. The total duration of treatment was 18 months. Endometrial biopsies were performed before treatment and between day 18 and day 24 of the 18th month of HRT. RESULTS: Of the 336 patients selected, 317 had a biopsy at inclusion. Of them, 244 patients (124 in the CA group and 120 in the P group) were suitable for evaluation for analysis at the 18th month. Insufficient sampling occurred in 33.9% in the CA group and 60% in the MP group (probably atrophic). No case of hyperplasia could be reported in both groups. The endometrium was atrophic in 19.5 versus 27.1%, proliferative in 3.7 versus 8.3% and secretory in 76.8 versus 62.5% in CA and MP groups, respectively. It was possible to see histological differences induced by the two progestins. The CA endometria showed fewer glands lined by a cubo-cylindrical epithelium, with an edematous stroma, compared to the MP endometria which had more glands lined by a cylindrical epithelium, stroma being poorly edematous. These figures varied in intensity due to the length of progestative impregnation, predecidualization occurring later in the CA group, with distended capillaries. CONCLUSIONS: These results show that CA 10 mg/day is a powerful progestin compared to MP 200 mg/day, on weakly estradiol-primed endometria, giving a molecule-specific histological aspect with a good endometrial safety.

Chlormadinone acetate versus micronized progesterone in the sequential combined hormone replacement therapy of the menopause.

OBJECTIVE: The efficacy and safety of chlormadinone acetate (CA) versus micronized progesterone (P) were assessed in non-hysterectomized postmenopausal women. MATERIALS AND METHODS: This was a multicenter, randomized, parallel group study with a 6-month double-blind period followed by a 12-month open period. Patients were randomized to receive every month during 18 months percutaneous 17 beta-estradiol (E(2)) 1.5 mg/day from Day 1 to 24 of treatment cycle, combined from Day 11 to 24 to either CA 10 mg/day (n=167) or P 200 mg/day (n=169). Endometrial biopsy (EB, main analysis criterion) was performed at baseline, and at Day 18-24 of the 6th and 18th cycles. RESULTS: At Month 6, EB did not evidence any hyperplasia. EB were inadequate for assessment in 24.5% and 47.5% of patients in the CA and MP groups, respectively. CA was found to be as protective as P (96.3% and 92.0% of success). However, the hormonal status of the endometrium differed (P<0.001): a secretory endometrium was found in 81.5% of the CA patients, compared to 50.7% in the P group. These transformations resulted in predictable, cyclic bleeding in 94.5% of the CA patients, compared to only 62.3% of the P patients (P=0.0001). Unscheduled bleeding, spotting and/or metrorrhagia, were more frequent under P than under CA (17.9% and 13.7%, respectively). The beneficial effects on hot flushes were more important in the CA group than in the P (P<0.001). At Month 18, the biopsy and clinical results were similar to those obtained at Month 6. The safety profile, particularly the lipid one, was similar in both groups, except for drowsiness and dizziness, which were significantly more frequent under P than under CA. CONCLUSION: The progestative effects of CA on the endometrium and on menopause-related symptoms were at least as good as those of P. Moreover, CA resulted more often than P in secretory effects, and in satisfying bleeding patterns.

Endometrial safety and tolerability of AERODIOL(R) (intranasal estradiol) for 1 year.

OBJECTIVE: the purpose of this study was to assess the endometrial safety and patient acceptability of a pulsed estrogen therapy provided by S21400 (intranasal 17 beta-estradiol) in the treatment of postmenopausal symptoms. DESIGN: postmenopausal women (n=408) entered an open-label, community based, multicentre trial. Patients received S21400 plus sequential (>90% of patients) or continuous progestogen. Treatment was initiated with a standard daily dose of 300 microg but dose adaptation was possible every 3 months from 150 to 600 microg daily. Endometrial biopsies were performed at entry and at 12 months, and bleeding patterns were recorded at 3-monthly intervals throughout the trial. RESULTS: 71% of patients received 300 microg per day S21400 throughout the study, 3% had their dose decreased, 19% had their dose increased and 7% had their dose both decreased and increased. Three hundred and eleven biopsies were obtained after 12 months of treatment, there were no cases of endometrial hyperplasia. The 95% confidence interval [CI] for the rate of incidence was 0-1.2%. Cyclical bleeding occurred in 82% of sequential treatment cycles. Unexpected bleeding occurred in 5% of the treatment cycles. Presence of unexpected bleeding varied according to the treatment regimen, 15 and 4% of the cycles with combined continuous and sequential regimen, respectively. Unexpected bleeding was mostly spotting. Nasal treatment was well accepted. Nasal symptoms (itching sensation, rhinorrhea and sneezing) were mostly mild in intensity and they led to treatment withdrawal in approximately 3% of patients. The rate of treatment continuation was 85% at 1 year. CONCLUSIONS: S21400, in combination with continuous or sequential progestogen, exhibits good endometrial safety and patient acceptability in postmenopausal women.

[Preliminary evaluation of a histocytological association predicted in a single sample of uterine exploration via a modified pipette: Mark II pipette]. / Evaluation préliminaire d'une association histo-cytologique prélevée en un seul geste pour exploration endo-utérine par une pipelle modifiée: la pipelle Mark II.

The pipelle has become the sampling tool of choice for histological detection of endometrial adenocarcinoma. Some practitioners still prefer cytological sampling. The distal extremity of the pipelle has been modified to allow removal of both types of samples simultaneously. The aim of this preliminary study was to evaluate the feasibility of this method. A positive agreement between histological and cytological results was found in 10 out of the 12 cases examined, among which an adenocarcinoma was positively detected. A prospective study on larger size must confirm these first findings. The new modified pipelle (Pipelle Mark II) offers a possible approach for reduction of false negatives.

Trophic effects on testes in paraplegics.

Testicular biopsies and hormone profiles were obtained from 23 paraplegic patients who had sustained a complete spinal cord section. The hormone profiles were normal, but patients with a spinal lesion including the T10-L2 metameres showed a particular pattern of germinal cell abnormalities. The atrophy is multifactorial, but may well include destruction of the sympathetic innervation of the testis by the lesion.

Profiles of plasma estrogens, progesterone and their metabolites after oral or vaginal administration of estradiol or progesterone.

Doses of 100 mg of micronized progesterone (P) and of 0.5 mg of micronized estradiol (E2) were administered vaginally and orally, respectively, in the early follicular phase of the menstrual cycle in six premenopausal women. In the second cycle, the same doses were administered in the same subjects, orally for P and vaginally for E2. Serial blood samples were collected and the following steroids were assayed by highly reliable techniques: P, E2, estrone (E1), deoxycorticosterone (DOC), 5 alpha- and 5 beta-pregnanolone and the sulfates of E1, E2, and DOC. Circulating P and E2 levels were higher after vaginal than after oral administration, while those of E1 were similar after either route. Metabolites of P (DOC, DOCS and pregnanolone) were higher after oral administration. Concerning estrogen sulfates, E1S concentrations were similar whichever the route, while those of E2S were lower after oral than after vaginal administration. This study has confirmed that metabolism of ingested P and E2 occurs mainly in the intestine. Moreover, P was predominantly metabolized to 5 alpha-reduced derivatives, whatever the route of administration. In view of the metabolic pathways which are operative and of the peripheral plasma levels which were found, the vaginal route appears to be more adequate than the oral one for hormone replacement therapy.

Androgen and 19-norsteroid profiles in human preovulatory follicles from stimulated cycles: an isotope dilution-mass spectrometric study.

Follicular fluid was aspirated from preovulatory follicles of women under ovarian stimulation for in vitro fertilization and analyzed by a highly specific technique based on gas chromatography-mass spectrometry associated with stable isotope dilution. 19-Nortestosterone and 19-norandrostenedione were identified and quantified for the first time in human follicular fluid. There was a strong positive correlation between 19-nortestosterone and estradiol-17 beta and between 19-norandrostenedione and estrone concentrations, thus indicating a common cellular origin. The accumulation of 19-norsteroids in follicular fluid confirms that they are weakly active intermediates in the multistep enzymatic conversion of androgen to estrogen. Testosterone concentrations were significantly lower than those obtained by radioimmunoassay; cross-reaction with substantially higher levels of 19-nortestosterone seems to be at the origin of this discrepancy. Androstenedione concentrations were similar to those reported in the literature and it was therefore confirmed that an estradiol/androstenedione concentration ratio above 20 is favourable for oocyte cleavage. Other and some newly estimated androgens are: testosterone sulfate, 5-androstene-3 beta, 17 beta-diol 3-sulfate and disulfate, dihydrotestosterone sulfate, epitestosterone, 19-hydroxyandrostenedione, 5 alpha-androstane-3 alpha, 17 beta-diol, 5 alpha-androstane-3 beta, 17 beta-diol, 5 alpha-androstane-3,17-dione and androsterone. Dehydroepiandrosterone sulfate was by far the most abundant androgen in this type of follicles.

Delayed appearance of plasmatic chorionic gonadotropin in pregnancies after in vitro fertilization and embryo transfer.

The detection of pregnancy through the rise of human chorionic gonadotropin hormone secretion, on maternal plasma level, has been studied in normally developed pregnancies following in vitro fertilization and embryo transfer (IVF-ET), and compared with two other groups of pregnancies, the first group being pregnancies following artificial insemination with donor semen (AID) in spontaneous cycles ("AID group") and the second group being pregnancies following in vivo fertilization in a stimulated cycle ("stimulated group"). The day of human chorionic gonadotropin detection level significant for pregnancy (Dd) has been first defined and then determined for each pregnancy. Thereafter, mean levels for Dd (Dd) have been compared for each pregnancy group. It has been found that in pregnancies following IVF-ET, Dd is 12.05 +/- 0.8 days after ovulatory stimulus, which is delayed in comparison with spontaneous cycle pregnancies (Dd = 9.5 +/- 1.0) and with stimulated cycle pregnancies (Dd = 8.0 +/- 1.5). The hypothesis to explain this observation is then discussed.

Effects of freezing and thawing on mammalian oocyte.

In an attempt to study the deleterious effects which occur during the freezing and thawing of mammalian oocytes, we developed a cryomicroscope controlled by digital programmable equipment. The program permits any cooling rate between 0.1 and 60 degrees C/min with a precision of 0.6 degrees C. Using a precooled stage, it is possible to obtain rapid cooling (100 degrees C/min). The maximum thawing rate is about 60 degrees C/min. A copper-- constantan microthermocouple allows precise measurement of the specimen temperature. All information (specimen, temperature of the specimen, date, hour, and minutes) is recorded at the same time on photographic film by a camera fitted with a " Recordata Back" and a motor drive which allows three frames per second. Our preliminary results show that: (1) rapid cooling yields a supercooling with simultaneous extra- and intracellular crystallization; (2) slow cooling with seeding at -8 degrees C gives an extracellular crystallization which is achieved by -9 degrees C, followed by an extracellular recrystallization occurring at almost -8 degrees C which alters the morphology of the oocyte and the zona pellucida, without any visible intracellular crystallization; (3) during continued slow cooling the oocytes dehydrate without any intracellular freezing; and (4) during rewarming a partial rehydration of the cell occurs with a swelling of the oocytes to their original volumes after the thawing has been achieved.

Pregnancy of the wife of a complete paraplegic by homologeous insemination after an intrathecal injection of neostigmine.

A case of successful pregnancy following artificial insemination following intrathecal neostigmine injection in the wife of a complete traumatic paraplegic (T7-T8 to T11-T12) is described.