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Biomedical subjects

R Rolland

Publications and source records attributed to R Rolland.

At least 55 records · Page 3Linked to original sources

Endometrial epithelial cells in peritoneal fluid during the early follicular phase.

Peritoneal fluid (PF) was obtained during the early follicular phase in 24 women at laparoscopy as part of infertility investigation. The cells present in PF were pelleted and cultured. Developing endometrial epithelial cell colonies were identified in 19 women (79%). Identification of these cell colonies was facilitated using the monoclonal antibody BW 495/36 as specific marker. The number of endometrial epithelial cell colonies showed a large variation (1 to 200 or more PF sample). No significant distinction in incidence and number of cell colonies was found between women with minimal (n = 11) and without endometriosis (n = 12). A significant correlation with number of cell colonies was found in women with infertility and no mechanical and male infertility factors. These data indicate that retrograde transport of viable endometrial cells during menstruation occurs in most women with patent tubes. Implications of the results for the relation between retrograde menstruation, endometriosis, and infertility are discussed.

Cells, Cultured↗

Retrograde seeding of endometrial epithelial cells by uterine-tubal flushing.

OBJECTIVE: To estimate the amount of endometrial epithelial cells in peritoneal fluid (PF) after uterine-tubal flushing (40 mL) throughout the menstrual cycle. DESIGN: We cultured the cell pellet of flush medium present in the peritoneal cavity. SETTING: University Hospital Nijmegen, The Netherlands. PATIENTS: Ninety-two women with various infertility-related factors. INCLUSION CRITERIA: (1) ovulatory cycle, (2) patent tubes, and (3) no adhesions. INTERVENTIONS: None MAIN OUTCOME MEASURE(S): The number of developing epithelial cell colonies were counted after 7 days. We started to record the amount of flush medium recovered during the study. RESULTS: The amount of flush medium recovered was positively correlated with the presence of endometriosis (P = 0.017). Endometrial epithelial cells were identified in 85 flush media (92%). The number of epithelial cell colonies varied from 0 to 100 or more and was higher when flushing was performed during the early follicular phase (P less than 0.01). High estradiol-17 beta and progesterone levels in culture medium did not change the number of developing cell colonies. Methylene blue significantly reduced the number of cell colonies (P = 0.002). CONCLUSIONS: Uterine-tubal flushing results in varying numbers of endometrial epithelial cells in PF. Methylene blue adversely affects the growth potential of these cells.

Cell Division↗

Vero cells stimulate human sperm motility in vitro.

OBJECTIVE: To study human sperm motility in a coculture system for in vitro fertilization (IVF). DESIGN: We studied the viability and motility (percentage and curvilinear velocity) of human spermatozoa after incubation in: (1) medium 199 supplemented with fetal calf serum (M199/FCS) together with Vero cells; (2) Vero cell conditioned M199/FCS; (3) M199/FCS supplemented with Vero cell extract; and (4) some control media. In a second experiment, FCS was substituted by sera from different IVF patients. SETTING: Semen samples were obtained from the fertility laboratory of the St. Radbound Hospital, Nijmegen, The Netherlands. PATIENTS: Twelve men of couples with fertility problems. INTERVENTIONS: None. MAIN OUTCOME MEASURE: The motility parameters were determined with a computerized motility-analyzing system, after 24 hours' incubation at 37 degrees C and 5% CO2. Viability was determined after eosin Y staining. RESULTS: Sperm viability was the same in all media. In the coincubation system, the Vero cell-conditioned medium, and the experiment with human sera, the sperm motility parameters were higher (P less than 0.005) than in the control media. Vero cell extract did not have this positive effect. CONCLUSIONS: Coincubation has a positive effect on sperm motility and may be beneficial to IVF, intrauterine insemination, and artificial insemination.

Animals↗

High CA-125 concentrations in peritoneal fluid of normal cyclic women with various infertility-related factors as demonstrated with two-step immunoradiometric assay.

OBJECTIVE: To determine CA-125 concentrations and total amounts in peritoneal fluid (PF) of women with various infertility-related factors throughout the menstrual cycle. DESIGN: Peritoneal fluid was obtained at laparoscopy. CA-125 was determined using the assessed two-step immunoradiometric assay (IRMA) which, in contrast to the one-step IRMA, gives valid results. SETTING: University Hospital Nijmegen, Nijmegen, The Netherlands. PATIENTS: One hundred six infertile women with a regular and ovulatory cycle were included. INTERVENTIONS: None. MAIN OUTCOME MEASURE(S): The mean PF CA-125 concentration and total amount were significantly lower during the luteal phase as compared with other phases of the menstrual cycle. No correlation was found with the presence or absence of endometriosis, adhesions, a male and/or cervical mucus infertility factor, and with patent or closed fallopian tubes. RESULTS: Peritoneal fluid CA-125 concentrations varied from 630 to 12,000 arbitrary units/mL (mean +/- SD = 3,437 +/- 2,286). Total PF CA-125 amounts (concentration x PF volume) varied from 1,760 to 13,300 arbitrary units (mean +/- SD = 30,219 +/- 26,841). CONCLUSIONS: CA-125 secretion into the abdominal cavity varies during the menstrual cycle. Retrograde menstruation is not the main source of CA-125 in PF.

Antigens, Tumor-Associated, Carbohydrate↗

Lactation inhibition by the dopamine agonist CV 205-502.

In an open pilot study with a parallel group design 30 bottle feeding women were randomly assigned in a two to one ratio to receive either the new dopamine agonist CV 205-502 or bromocriptine for lactation inhibition. Ten women who intended to breast feed served as normal controls. All treated women reached prepregnant prolactin levels within 72 h with once-daily 0.075 mg of CV 205-502 or twice-daily 2.5 mg of bromocriptine, at starting doses of 0.05 mg and 2.5 mg respectively. Fifteen of the 20 women treated with CV 205-502 reported breast symptoms, 50% occurring on days 3 and 4 of treatment. Three of the 10 women treated with bromocriptine had breast symptoms between days 2 and 28. Overall efficacy and tolerance in the two groups was very good. Side effects did not differ between the groups, with the exception of pulse rate in the standing position, which was significantly higher in the bromocriptine treated group than in either the CV 205-502 group (P = 0.02) or the breast feeding group (P less than 0.01). The coagulation tests (fibrinogen, AT III, PTT and APTT) showed no significant differences between the three groups.

Administration, Oral↗

Pulsatile secretion of luteinizing hormone and prolactin in lactating and nonlactating women and the response to naltrexone.

To investigate the mechanisms responsible for the postpartum suppression of reproductive function, LH and PRL levels were determined at 10-min intervals for 8 h in 10 lactating women, in 10 nonlactating women treated with bromocriptine, and in 5 nonpuerperal women. The lactating women were studied on postpartum day 7 (n = 5) or between days 28 and 35 (n = 5). All nonlactating women were studied on day 7. Five of them were treated with the opioid antagonist naltrexone to evaluate the role of endogenous opioids. By using a specific LH assay which does not cross-react with hCG, we were able to measure pulsatile LH secretion in the early puerperium for the first time. On day 7, LH levels were below the detection limit in both lactating and nonlactating women, hence no pulses could be detected. Chronic opioid blockade in bromocriptine-treated non-lactating women did not result in increased LH levels. Pulsatile LH secretion was present between days 28 and 35 of lactation, although pulse amplitude (P less than 0.05) and mean LH level (P less than 0.05) were lower than in nonpuerperal women. Mean LH in lactating women was negatively correlated with mean PRL (-0.87, P less than 0.05). Deconvolution analysis of the PRL responses to suckling revealed that PRL was secreted in distinct bursts, separated by intervals of secretory quiescence. Mean duration of PRL bursts was 40 +/- 4 min and the maximum secretory rate was reached around the end of suckling. We conclude that pulsatile LH secretion is completely suppressed during early lactation and partially suppressed during late lactation. Because the duration of suckling was similar, the relatively strong suppression during the early puerperium must be due to additional inhibitory factors. It has been suggested that endogenous opioids are involved in this process, but our results in puerperal women do not support this hypothesis.

Adult↗

Nafarelin versus danazol in the treatment of endometriosis.

A double-blind, double-dummy, 6-month multicenter study comparing the effects of nafarelin and danazol in the treatment of endometriosis was completed recently by investigators from 13 institutions in seven European countries. The 194 patients (aged 18 to 45 years) selected for the study were divided into two groups. One group received nafarelin, 200 micrograms twice daily, and placebo capsules identical to danazol, twice daily. The other group received danazol capsules, 200 mg twice daily, and a placebo nasal spray identical to nafarelin, twice daily. A comparison of decreases in both groups of American Fertility Society endometriosis scores and in severity scores developed by the investigators indicates that nafarelin and danazol are equally effective in the treatment of endometriosis.

Adolescent↗

Prevention of postoperative adhesions by tissue-type plasminogen activator (t-PA) in the rabbit.

The fibrinolytic system of the peritoneum is important in the pathogenesis of adhesions. Plasminogen activator activity is depressed by serosal types of injury, which cause ischemia. Ischemic peritoneum induces fibrinous adhesions. When local fibrinolytic activity is impaired, persistence of fibrin and organisation of the fibrinous adhesions may occur. Peritoneal adhesion in rabbits were created by drying the serosa of the uterine horns and by introducing a small amount of blood intraperitoneally. In one group of rabbits tissue-type plasminogen activator (t-PA) was given intraperitoneally at the end of the operation: another group served as controls. 48 rabbits were operated on. The animals were killed 3 h, 1, 3 and 7 days postoperatively for evaluation of adhesions. All animals of the control group had adhesions. The animals of the t-PA group had less adhesions than the animals of the control group.

Animals↗

Lactation inhibition by a single injection of a new depot-bromocriptine.

Recently, long-acting injectable forms of bromocriptine have become available for the prevention of lactation. The first developed depot-form was very effective, but had the disadvantage of a slowly metabolized carrier. we investigated the pharmacokinetics, efficacy, tolerance and safety of 40 and 50 mg of a new rapidly eliminated depot-form in 61 postpartum women. Bromocriptine rapidly increased after injection and prolactin was effectively suppressed during the study-period of 60 days. Overall efficacy was very good or good in 98% and no rebound lactation occurred. Sixteen women experienced side effects. Tolerance at the injection site was good and safety tests did not show abnormalities. There were no differences between the two dosages. We conclude that this new depot-bromocriptine is a safe, well tolerated and effective drug in the suppression of prolactin and the prevention of post-partum lactation.

Adult↗

Postpartum return of pituitary and ovarian activity during lactation inhibition with the new dopamine agonist CV 205-502 and during normal lactation.

The new nonergot dopamine agonist CV 205-502 appears to be a promising alternative in the treatment of hyperprolactinemia. Regarding the potential use of CV 205-502 in infertility practice, we studied the influence of CV 205-502 on the return of endocrine fertility parameters during the physiological hyperprolactinemia of the puerperium. The resumption of pituitary and ovarian activity in 18 CV 205-502 treated women was compared with that in 10 bromocriptine-treated women. LH was measured by a new specific assay, which does not cross-react with hCG. This assay was also used in a second part of the study in which the pituitary function of 10 breast-feeding women was investigated. Both dopamine agonists adequately suppressed PRL. Pituitary secretion returned in the second week and was initially characterized by a high FSH/LH ratio. There were no major differences between CV 205-502 and bromocriptine. Ovulations occurred from day 18 on. The PRL rebound at the end of treatment seemed to play a role in the ovulation process. An acute increase of PRL just before midcycle was able to prevent ovulation. Breastfeeding women showed a delayed return of pituitary secretion: after a hypogonadotropic period, FSH returned in the third week and was followed by a period with a high FSH/LH ratio and follicular inactivity.

Aminoquinolines↗

Human choriogonadotropin (hCG): comparisons between determinations of intact hCG, free hCG beta-subunit, and "total" hCG + beta in serum during the first half of high-risk pregnancy.

We have studied the concentrations of intact human choriogonadotropin (hCG) and the free hCG beta-subunit in blood samples from singleton pregnancies at risk for habitual or threatened abortion. The samples were obtained weekly between the 6th and 12th weeks and in the 14th and 16th weeks of gestational age. The concentrations of intact hCG, of the free hCG beta-subunit, and of "total" hCG (i.e., intact hCG and the free hCG beta-subunit: hCG + beta) were measured in serum by specific immunoassays. The distributional statistics (the 5th, 50th, and 95th percentiles) of "total" hCG + beta and of intact hCG showed very similar patterns, whereas the response curves for the free hCG beta-subunit showed very much lower serum concentrations. From these data we also estimated distributional statistics of the percent molar ratios of free hCG beta-subunit to intact hCG. We conclude that (a) the relatively small proportion of free hCG beta-subunit in serum during the first half of singleton pregnancy is far too low to interfere with the applied "total" hCG assay, as compared with the serum values obtained for intact hCG, and (b) the percent molar ratios of free hCG beta-subunit to intact hCG, or to "total" hCG + beta, never exceeded 1.0% throughout the period of pregnancy studied.

Abortion, Threatened↗

Successful pregnancy after ZIFT in a patient with congenital cervical atresia.

We report the third patient with a successful pregnancy following operative correction of congenital cervical atresia. The pregnancy was achieved through zygote intrafallopian transfer (ZIFT). Although stenosis of the newly formed cervical canal causes considerable morbidity, therapy should be aimed at the creation of a conduit for menstrual blood and for possible future pregnancy. New techniques in assisted reproduction such as in vitro fertilization-embryo transfer, gamete intrafallopian transfer, and ZIFT increase the chances of pregnancy in patients with congenital cervical atresia. A hysterectomy, as advocated until very recently, should not, in our opinion, be the first treatment of choice.

Adolescent↗

ETN-1: a new human endometrial carcinoma cell line producing ascites and distant metastases in nude mice.

A human carcinoma cell line (ETN-1) has been established from a skin metastasis of a moderately differentiated adenocarcinoma of endometrial origin. The cell line has been so far maintained for 27 months through 55 passages, growing as a monolayer as well as in 3-dimensional clusters with a population doubling time of 72 hr. The number of chromosomes per cell varied from 39 to 107 (average number 61.0 +/- 19.8), with a modal number of 46-48. Seven clonal marker chromosomes were detected. Flow cytometric analysis revealed a population of pseudo-tetraploid cells (DNA index 2.1) next to a pseudo-diploid population (DNA index 1.1). The epithelial character of the cells was confirmed by a positive immunocytochemical reaction using monoclonal antibodies (MAbs) to different keratins, the epithelial cell markers BW 495/36 and HMFG-2, as well as by the presence of many junctional complexes. The tumour cells retained a positive reaction with the anti-ovarian carcinoma OV-TL 3, OV-TL 10 and OC 125 MAbs, although the reaction was markedly diminished in comparison with the original tumour. Tumour cells inoculated subcutaneously in nude mice produced well differentiated adenomatous tumour nodules with formation of glandular lumina and basal lamina. Tumour cells injected intraperitoneally produced malignant ascites and regional as well as distant metastases of adenomatous character.

Adenocarcinoma↗

Endometriosis: treatment with gonadotropin-releasing hormone agonist Buserelin.

Fifty-one women with pelvic endometriosis were treated with the gonadotropin-releasing hormone agonist (GnRHa) Buserelin (Hoechst Holland N.V., Amsterdam, The Netherlands) 300 micrograms three times a day intranasally for 6 months. Forty-nine women completed treatment; 42 were available for 6 months of follow-up following treatment. Symptoms showed prompt and significant improvement. Follow-up after treatment revealed persistent relief from dysmenorrhea and dyspareunia in, respectively, 58.6% and 88.2% of the women, whereas pelvic pain returned to pretreatment scores. Serum estradiol (E2) was suppressed to predominantly early follicular phase concentrations. Laparoscopy at the end of therapy showed significant reduction of scores for implants only. There was no relation between the degree of E2 suppression during therapy and the improvement of symptoms or the reduction of endometriosis. Statistical analysis in 22 infertile patients, of whom 7 conceived during follow-up, revealed no differences in E2 levels during therapy, improvement of symptoms, or reduction of endometriosis. Buserelin appears to be safe, well tolerated, and effective in the management of endometriosis and associated complaints.

Adult↗

Return of gonadotropic function in postpartum women during bromocriptine treatment.

The rapid postpartum return of gonadotropic function during bromocriptine treatment was investigated through hormonal measurements in 15 women. Luteinizing hormone (LH) was measured by a novel and specific assay that does not cross-react with human chorionic gonadotropin (hCG). Prolactin was effectively suppressed by bromocriptine and hCG was cleared within 2 weeks. During the first week, both follicle-stimulating hormone (FSH) and LH were undetectable. The return of FSH and LH secretion occurred in the second postpartum week and initially was characterized by a high FSH/LH ratio. The causes of the 1-week delay in resumption of pituitary gonadotropin secretion are not known, but endogenous opioids may play a role. The possible relation between the high FSH/LH ratio and low LH pulse frequency also is discussed.

Bromocriptine↗