Female Volunteers Of Reproductive Age Research Articles

P–587 Highly purified human menopausal gonadotrophin (HP-hMG, Menopur) as a ready-to-use solution for injection in pre-filled pen is bioequivalent to HP-hMG powder for reconstitution

Abstract Study question Are serum FSH levels after single subcutaneous dosing of HP-hMG in a liquid formulation and a powder formulation bioequivalent? Summary answer The 90% CIs for the geometric mean ratios of serum FSH AUCt and Cmax were both within 0.8000–1.2500, thus the two formulations are bioequivalent. What is known already For several decades, HP-hMG (Menopur) has been used for the treatment of infertility; its efficacy and safety compared to other gonadotropins have been consistently demonstrated in several prospective, randomised controlled trials and meta-analyses (Deeks et al 2018; Bordewijk et al 2019). Menopur powder for reconstitution is available in multidose and single dose formulations. Up to 3 single dose vials (each containing 75 IU) may be dissolved into 1 mL solvent for administration. Recently, and for the first time, Menopur has been successfully formulated in a stable, ready-to-use solution for injection, which may be administered by a pre-filled pen. Study design, size, duration This was a randomised, two-way crossover, single dose, bioequivalence trial comparing Menopur liquid injected by pre-filled pen, with Menopur powder injected by conventional syringe and needle. The primary endpoints were AUCt and Cmax of baseline-adjusted FSH. Pituitary-suppressed, healthy women were randomised to receive one treatment sequence including a single subcutaneous injection of 450 IU Menopur liquid (600 IU/0.96 mL) and of 450 IU Menopur powder by two subcutaneous injections of 225 IU in 1 mL. Participants/materials, setting, methods Blood samples were collected pre- and post-dose, until 9 days after each injection. The PK parameters of FSH and hCG were assessed by noncompartmental methods with adjustment for endogenous pre-dose levels. Highly sensitive and specific electrochemiluminescence immunoassays were used for quantification and the LLOQ of the FSH and hCG assays were 1.47 mIU/mL and 0.5 mIU/mL, respectively, and the total validated CV was within 5% for both assays. Main results and the role of chance In total, 76 women were randomised and 56 completed the trial. The main reason for discontinuation was insufficient pituitary suppression prior to the second administration of HP-hMG. The mean FSH and hCG serum concentration-time profiles were comparable between the two HP-hMG formulations. The geometric mean ratios and 90% confidence intervals of FSH for HP-hMG liquid versus HP-hMG powder were 1.12 [1.0562; 1.1889] for AUCt and 1.17 [1.0946; 1.2490] for Cmax, showing that the two formulations were bioequivalent. Maximal serum FSH concentrations were reached at 18.19 h for HP-HMG liquid and 15.55 h for HP-hMG powder. In addition to FSH, the PK parameters for hCG were compared between the two HP-hMG formulations. The geometric mean ratios and 90% confidence intervals for HP-hMG liquid versus HP-hMG powder were 0.93 [0.86; 1.01] for AUCt and 0.94 [0.86; 1.02] for Cmax. There was no difference between the two groups in the incidence or severity of adverse events, and both preparations were well tolerated. Mild injection site reactions were less common after administration of HP-hMG liquid by a single injection compared to HP-hMG powder by two injections and were mostly related to pain and erythema after drug administration. Limitations, reasons for caution This bioequivalence study is based on the comparison of single dose administrations in healthy female volunteers of reproductive age. Wider implications of the findings: The new HP-hMG solution for injection in a pre-filled pen will deliver the efficacy and safety of Menopur in a convenient delivery device. Trial registration number NA

Dynamics of proinflammatory cytokine serum levels in patients with acute inflammatory diseases of pelvic organs in the early stages of conservative treatment

Dynamics of serum proinflammatory cytokines such as interleukins (IL)-1, -2 and -6, tumor necrosis factor alpha (TNFα) was examined in patients of reproductive age suffering from acute inflammatory diseases of the pelvic organs. Our study was carried out with female patients at hospital admission prior to therapy applied with conventional methods of treatment and improved treatment by using vaginal low-frequency laser in a constant continuous magnetic field. The data obtained were compared with serum cytokine level from 20 healthy female volunteers of reproductive age consulted on better contraception methods. It was found that cytokine profile of in patients with acute inflammatory processes in pelvic organs was characterized by a high level of proinflammatory cytokines. It was shown that patients receiving conventional treatment contained decreased level of serum IL-1, IL-2, IL-6 and TNFα displaying slight dynamics, which did not reach it in control group. This may contribute to ongoing inflammatory process, despite the positive clinical dynamics. In turn, imbalance of immune responses leads to a persistently impaired fertility in women and need to perform subsequent comprehensive rehabilitation measures. Moreover, patients applied with intravaginal low-frequency laser radiation in a constant magnetic field were found to contain serum TNFα 100 pg/ml observed in 59.7% of cases, IL-6 level was lower than 20 pg/ml (prevalent in control group) found in 54.2% of cases. Serum IL-2 level was decreased by 3.5-fold compared to baseline, whereas for IL-1β it was higher than 100 pg/ml in as few as 23.6% patients. Such temporal pattern of inflammatory markers with rapid significant decrease of serum proinflammatory cytokines in patients with preformed pathogenic factors can reduce probability of connective tissue formation and activate their own repair as well as regenerative events. The results obtained allow to wider use combined physical interventional factors for therapy of patients with acute inflammatory diseases of the pelvic organs.

Insulin resistance is associated with hirsutism in unselected reproductive-aged women.

Hirsutism affects 5%-10% of reproductive-aged women worldwide and exhibits clinical importance as a cutaneous manifestation of underlying hyperandrogenism. Racial and genetic factors play roles in manifestation of hirsutism, and the prevalence of hirsutism seems to be low in East Asians. However, the reference value of the modified Ferriman-Gallwey (mFG) score to diagnose hirsutism and the prevalence of hirsutism have not been determined in Korean populations to date. We aimed to investigate the distribution of the mFG score and establish its reference value for defining hirsutism and to examine its relationship with metabolic and reproductive traits in reproductive-aged Korean women. We enrolled 2139 female volunteers of reproductive age (15-39years). We recorded mFG scores from 0 to 4 on 9 different body locations (upper lip, chin, chest, arm, upper abdomen, lower abdomen, upper back, lower back and thighs). Hirsutism was defined as >95th percentile of mFG score. In addition, a 75-g oral glucose tolerance test was performed, and the homoeostasis model assessment of insulin resistance (HOMA-IR) was calculated. The mFG values of the 50th, 75th, 90th and 95th percentiles were 0, 1, 4 and 6, respectively. Therefore, the mFG score was indicative of hirsutism when the score was 6 or greater, which represents the 95th percentile. In the correlation analysis, total testosterone, free testosterone, fasting plasma insulin and HOMA-IR were positively correlated with mFG score (all Ps <0.05). Multiple linear regression analysis revealed that HOMA-IR (β=0.081) was positively associated with mFG score after adjustments for age, body mass index, total testosterone and the number of menses per year (P<0.001). In conclusion, setting the 95th percentile of the mFG score as normal, the reference value to define hirsutism was 6 in reproductive-aged Korean women. HOMA-IR was positively associated with the mFG score even after adjustment for biochemical hyperandrogenism.

First Evidence of Ovulation Induced by Oral LH Agonists in Healthy Female Volunteers of Reproductive Age

Two new low-molecular-weight LH agonists (Org 43553 and Org 43902) were shown to induce ovulation in preclinical experiments. Our objective was to assess the safety, pharmacokinetics, and pharmacodynamics of Org 43553 and Org 43902 when administered to healthy females. Org 43553 and 43902 studies were randomized, placebo-controlled, single-rising-dose first-in-human trials, which included 159 healthy female volunteers. Part 1 of the studies assessed the safety and pharmacokinetics. Part 2 evaluated the pharmacodynamics effect of a single oral dose of Org 43553 (25-900 mg) or Org 43902 (30-300 mg) to induce ovulation after the development of a large preovulatory follicle, whereas the endogenous LH surge was suppressed due to GnRH antagonist treatment while follicular development was supported with recombinant FSH. Org 43553 and 43902 were safe and well tolerated. Both compounds showed a fast absorption after oral intake, with peak concentrations reached within 0.5 to 1 hour. The elimination half-life of Org 43553 was 30 to 47 hours and that of Org 43902 was 17 to 22 hours. Ovulation induction confirmed by midluteal progesterone rise ≥15 nmol/L was proven in both studies, also when excluding subjects with an endogenous LH rise. The minimal effective dose for ovulation induction was 300 mg in both studies and resulted in an ovulation rate of 83% and 82%, respectively. These first proof-of-concept studies both demonstrated that a single oral intake of an low-molecular-weight LH agonist induces ovulation of the preovulatory follicle in pituitary-suppressed female volunteers of reproductive age.

Effects of tampons and menses on the composition and diversity of vaginal microbial communities over time

To investigate the influence of menses on the vaginal microbiota and determine whether tampons that differ in material composition influence these bacterial communities in different ways. A single-centre trial with randomised, complete block design. Procter & Gamble facility. Seven self-declared healthy, female volunteers of reproductive age. Volunteers used a pad and two types of tampons during the study, one product exclusively each month for three sequential menstrual cycles. During menses and once each mid-cycle, vaginal bacterial community composition was characterised by cultivation-independent methods based on pyrosequencing of V1-V2 variable regions of 16S ribosomal RNA genes. Changes in the species composition, abundance and diversity in vaginal bacterial communities over time and between treatments. The vaginal microbiotas of all seven women were dominated by Lactobacillus spp. at mid-cycle, and the compositions of those communities were largely consistent between cycles. Community dynamic patterns during menses varied considerably and were more or less individualised. In three of the seven women the community diversity during pad use was significantly different from at least one tampon cycle. Changes in the composition of the vaginal microbiota during menses were common, but the magnitude of change varied between women. Despite these changes, most communities were capable of resuming a composition similar to previous mid-cycle sampling times following menstruation. Overall we conclude that the two tampons tested do not significantly impact the vaginal microbiota in different ways; however, larger studies should be performed to confirm these findings.

Intrauterine devices and uterine peristalsis: evaluation with MRI

Intrauterine devices (IUDs) have been viewed as an effective form of contraception. However, the mechanism by which IUDs disturb fertility remains controversial. This study aimed to evaluate the effects of IUDs on uterine contractility using cine MR. Eleven healthy female volunteers of reproductive age bearing IUDs and 12 women not bearing IUDs were evaluated during the periovulatory phase. MR images were obtained with a 1.5-T magnet, acquiring 60 serial images every 3 s via half-Fourier acquisition single-shot turbo spin echo to be displayed on cine mode. Assessments were based on (a) the presence of peristaltic waves, (b) the frequency and direction of peristaltic waves and (c) the extent of peristaltic waves. Static images were evaluated for thickness of the junctional zone (JZ) and myometrium. A fundo-cervical (FC)-directed peristaltic wave was identified in 4 of 11 IUD-bearing subjects and in only 1 of 12 subjects from the control group. FC waves extended through more than half of the thickness of the myometrium. Peristaltic frequency in IUD users (5.0/3 min) was less than that of the control group (6.5/3 min). The JZ and myometrium were significantly thicker in IUD users. FC-directed waves were more often observed in IUD-bearing subjects, which might explain the inhibition of active sperm transport.

Effects of a single Silastic® contraceptive implant containing nomegestrol acetate (Uniplant) on endometrial morphology and ovarian function for 1 year

Purpose This study was undertaken to evaluate the effects of a subdermal implant containing nomegestrol acetate (Uniplant) on endometrial histology and ovarian function. Methods Twenty healthy female volunteers of reproductive age were included and completed a menstrual diary throughout the study. Hysteroscopy, transvaginal sonography and blood sampling were performed prior to implant insertion (control cycle) and following 6 and 12 months of Uniplant use. Transvaginal sonography was performed every other day from Day 8 of the cycle up to the obtainment of sonographic evidence of a 12-mm follicle, then every day until the obtainment of sonographic evidence of follicular rupture and thereafter every other day until the next menstrual bleeding. Blood samples were taken for the measurement of estradiol, follicle-stimulating hormone, luteinizing hormone and progesterone on the same days on which transvaginal sonography was performed. The implants were removed after 1 year. Results Twenty percent of cycles were ovulatory, and 80% were anovulatory. The development of persistent nonluteinized follicle occurred in 40% of all cycles studied, inadequate luteal phase occurred in 20% of cycles and no follicular development occurred in 40%. Endometrial thickness remained below 8 mm in all cycles studied. Alterations in endometrial vascularization were observed in all treated cycles. Conclusion Our results suggest that this long-acting contraceptive method affects follicular growth and endometrial vascularization, disrupts endometrial architecture and leads to inadequate luteal phase.

Oral contraceptives and uterine peristalsis: Evaluation with MRI

To evaluate the effects of oral contraceptives (OCs) on uterine contractility using cine MRI, and correlate the kinematic findings with the static findings. Healthy female volunteers of reproductive age (23 taking OCs, and 15 not) were evaluated at mid-cycle. MR images were obtained with a 1.5T magnet, with 60 serial images taken every three seconds by half-Fourier acquisition single-shot turbo spin echo (HASTE) to be displayed in cine mode. Assessments were based on the 1) detectability of uterine peristalsis in cine mode, 2) peristaltic frequency and direction, 3) thickness of the endometrium, myometrium, and junctional zone (JZ) on T2-weighted images (T2WIs), and 4) intensity of the myometrium and cervical mucus on T2WIs and T1-weighted images (T1WIs), respectively. Uterine peristalsis was markedly decreased in the OC users. The endometrium and JZ were significantly thinner, and the myometrium was thicker in the OC users compared to controls. The signal intensity of the myometrium and cervical mucus was significantly higher in the OC users than in controls. OCs markedly suppressed uterine peristalsis at mid-cycle, and the uterus displayed a globular configuration with a thin JZ that may be related to decreased uterine peristalsis in OC users.

Pharmacokinetic and pharmacodynamic characteristics of ganirelix (Antagon/Orgalutran ). part II. dose-proportionality and gonadotropin suppression after multiple doses of ganirelix in healthy female volunteers

Objective: To assess the dose-proportionality and pharmacodynamic properties of multiple doses of ganirelix (Antagon/Orgalutran; NV Organon, Oss, the Netherlands). Design: Randomized, parallel, pharmacokinetic, and pharmacodynamic study. Setting: Phase I clinical research unit. Patient(s): Three groups of 15 healthy female volunteers of reproductive age. Intervention(s): Subcutaneous injections of 0.125 mg, 0.25 mg, or 0.50 mg of ganirelix were given once daily for 7 days. Blood samples were taken to assess serum ganirelix, LH, FSH, and E 2 concentrations. Main Outcome Measure(s): Pharmacokinetic parameters and hormone suppression. Result(s): Steady-state levels were reached between days 2 and 3. Peak concentrations, which occurred approximately 1 hour after dosing, increased in a dose-proportional manner and averaged 5.2 ng/mL, 11.2 ng/mL, and 22.2 ng/mL for the 0.125-mg, 0.25-mg, and 0.50-mg doses, respectively. Corresponding mean values for the area under the curve over one dosing interval (24 hours) were 33 ng · h/mL, 77.1 ng · h/mL, and 137.8 ng · h/mL, respectively. After the last 0.25-mg dose of ganirelix, serum LH, FSH, and E 2 concentrations were maximally decreased (by 74%, 32%, and 25% at 4 hours, 16 hours, and 16 hours after injection, respectively). Serum hormone levels returned to pretreatment values within 2 days after the last injection. Conclusion(s): The pharmacokinetics of ganirelix were dose-proportional within the dose range studied. Multiple injections resulted in immediate suppression of gonadotropins, which was rapidly reversed after treatment discontinuation.

Pharmacokinetic and pharmacodynamic characteristics of ganirelix (Antagon/Orgalutran ). part I. absolute bioavailability of 0.25 mg of ganirelix after a single subcutaneous injection in healthy female volunteers

Objective: To assess the absolute bioavailability of ganirelix (Antagon/Orgalutran; NV Organon, Oss, the Netherlands) after a single SC injection. Design: Randomized, crossover, pharmacokinetic study. Setting: Phase I clinical research unit. Patient(s): Nineteen healthy female volunteers of reproductive age. Intervention(s): Two separate injections of 0.25 mg of ganirelix were given, one subcutaneously and one intravenously, with a washout period of 1 week between injections. Blood samples were taken for assessment of serum ganirelix concentrations, and blood pressure, heart rate, and adverse events were monitored. Main Outcome Measure(s): Pharmacokinetic parameters. Result(s): Fifteen subjects were evaluated. The mean concentration-time profile after SC administration was comparable to that after IV administration. The mean (±SD) peak concentration and time of occurrence after SC administration were 14.8 ± 3.2 ng/mL and 1.1 ± 0.3 hours, respectively. The mean (±SD) half-lives after IV administration and SC administration were highly similar (12.7 ± 3.7 hours and 12.8 ± 4.3 hours, respectively). Mean (±SD) AUC 0–∞ (area under the concentration-time curve) values of 105 ± 11 ng/mL × hours and 96 ± 12 ng/mL × hours were calculated for IV administration and SC administration, respectively, resulting in an absolute mean (±SD) bioavailability of 91.3% ± 6.7%. Both treatments were well tolerated. Conclusion(s): Ganirelix is absorbed rapidly and extensively after SC administration, resulting in a high absolute bioavailability of >90%.

Dietary composition and ovarian function among Lese horticulturalist women of the Ituri Forest, Democratic Republic of Congo.

To assess dietary composition in relation to energetic status and ovarian function. An eight-month prospective study undertaken in 1989. Ituri Forest, northeast Democratic Republic of Congo. Sixty-four female volunteers of reproductive age (16-45), with a subsample of 30 for nutritional focal follows, 39 for analysis of salivary progesterone levels, and 18 for analysis of salivary oestradiol. Regular anthropometric measurements of heights, weights, and mid-triceps skinfolds, 204 h of nutritional focal follows when all foods prepared and consumed were recorded and weighed, analysis of dietary composition using African food tables, collection of saliva samples every-other day for radioimmunoassay of salivary oestradiol and progesterone. The primary staple in the Lese diet is manioc, consumed at nearly every meal, but the diet is subject to seasonal fluctuations. Important seasonal crops are groundnuts and rice. Carbohydrates comprise 64% of the diet, protein 14% and fat 22%, but seasonal fluctuations in food items create significant differences in protein content (P = 0.007). Energy intake falls by 25% in the hunger season. Lese energy balance reflects this lack with women losing a mean 7% of total body weight. The mean BMI for Lese women at the beginning of the study was 22.8 but, during the hunger season, the BMI for 20% of women fell to < 18.5, indicative of chronic energy shortage. Mean levels of Lese salivary oestradiol and progesterone are chronically and significantly lower than healthy, Western controls (P = 0.0001 for progesterone; P = 0.03 for follicular values of oestradiol, P = 0.0001 for mid-cycle values, and P = 0.0002 for luteal values). Mean salivary progesterone levels were significantly lower for those Lese women losing weight (P = 0.03), and significantly lower for weight-loss women when comparing levels at the beginning and end of the hunger season (P = 0.03). The relatively low-fat, high-fibre diet of the Lese appears healthy, but there are occasional seasonal deficiencies in protein-energy and micronutrient content, especially for pregnant and lactating women. Dietary composition may affect Lese levels of reproductive steroids, partly explaining the chronically low salivary oestradiol profile of this population.

Ovarian steroids modulate human monocyte tumor necrosis factor alpha messenger ribonucleic acid levels in cultured human peripheral monocytes

To determine whether tumor necrosis factor alpha (TNF-alpha) messenger ribonucleic acid (mRNA) levels in human peripheral monocytes are regulated by ovarian steroids. Human granulosa-luteal cells and cultured, activated human peripheral monocytes were subjected to Northern blot analysis for TNF-alpha mRNA. Academic research laboratory. Two human female volunteers of reproductive age and in the luteal phase of the menstrual cycle. Human granulosa-luteal cells produce TNF-alpha mRNA. Physiological levels of progesterone (P) and estradiol (E2) modulate TNF-alpha mRNA from peripheral blood monocytes with an apparent inverse relationship between steroid concentration and TNF-alpha message. Progesterone and E2 at physiological concentrations regulate TNF-alpha mRNA production. The P antagonist mifepristone (RU486) and the E2 agonist/antagonist tamoxifen modulate total TNF-alpha mRNA levels, suggesting involvement of specific receptors.

A pilot study on the assessment of a progesterone/estradiol sustained release as once-a-month-injectable contraceptive

A pilot study to assess the use of natural hormones in macrocrystalline sustained release system was undertaken in normal menstruating women. Progesterone at a dose of 100 mg in combination with 5 mg estradiol-17β aqueous macrocrystalline suspension (3ml) of defined particle size range (100–250 μm) were administered to five female volunteers of reproductive age, on day 5 of their normal menstrual cycles and then every 28 days consecutively for the next two months. Peripheral venous blood samples were obtained from the women three times a week for 60 days after the third injection for the measurement of serum progesterone, estradiol-17β, LH and FSH. The menstrual bleeding patterns were closely monitored during the study period. The results obtained indicate that the exogenous hormone administration produces blood levels similar to those observed during the luteal phase of the menstrual cycle. Follicular maturation as assessed by endogenous estradiol rise, above 150 pg/ml, occurred 29.7 days s.d. 6.4 after the injection. Ovulation as measured by progesterone levels above 5 ng/ml was documented 34.4 days s.d. 4.3 after the third injection. The bleeding patterns were regular though initially shorter but these increased progressively towards normal pattern during course of the study. The data suggest that progesterone/ estradiol —17β combination administered as an aqueous macrocrystalline suspension is capable of producing sustained ovulation inhibition and could be applied in the design of new once-a-month injectable contraceptives.