Otello Magrini

Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes

OBJECTIVE To describe the first birth achieved after intracytoplasmic sperm injection (ICSI) of cryopreserved human oocytes. DESIGN Case report. SETTING University of Bologna Hospital, Department of Obstetrics and Gynecology, Reproductive Endocrinology Unit, IVF and Infertility Center. PATIENT(S) One patient undergoing IVF. INTERVENTION(S) Transvaginal ultrasound-guided oocyte retrieval followed by oocyte freezing. Artificial preparation of the endometrium with E2 and P, oocyte thawing, and ICSI. RESULT(S) Four of 12 cryopreserved oocytes survived; using ICSI, 2 underwent normal fertilization but only 1 cleaved. One good-quality 4-cell embryo was transferred. A single gestation was confirmed by ultrasound at the 7th week. Amniocentesis was performed at the 16th week and demonstrated a normal female karyotype of 46,XX. After a normal pregnancy, a healthy female infant was born at the 38th week of gestation. CONCLUSION(S) The combination of ICSI and oocyte cryopreservation is a new tool in assisted reproductive technology.

Meiotic spindle recovery is faster in vitrification of human oocytes compared to slow freezing

OBJECTIVE To investigate spindle behavior during and after slow freezing at room temperature (RT) and vitrification at different temperatures. DESIGN Randomized, comparative study. SETTING University hospital. PATIENT(S) Patients undergoing IVF treatment volunteered for the study and donated part of their supernumerary oocytes. INTERVENTION(S) Metaphase II oocytes were divided into group A: slow freezing RT /thawing RT; group B: vitrification RT/warming RT; group C: vitrification RT/warming 37 degrees C; and group D: vitrification 37 degrees C/warming 37 degrees C. Spindle presence was evaluated at each step of the four procedures and in culture. MAIN OUTCOME MEASURE(S) Cumulative spindle recovery rate comparing warming phase of the three vitrification groups and culture phase among the four groups. RESULT(S) During warming, the three vitrification groups showed a significantly fast spindle recovery rate compared to the thawing of the slow freezing group. A progressively significant fast cumulative recovery rate was observed in the three vitrification groups by increasing the number of phases at physiological temperature (hazard rate = 2.68; 95% confidence interval 1.71-4.02). CONCLUSION(S) The present study demonstrates that spindle recovery is faster in vitrification than in slow freezing. These data support a possible protective effect of vitrification/warming at 37 degrees C on the meiotic spindle structure and, therefore, on the subsequent clinical outcome of the procedure, although comparative clinical studies are needed.

Postmenarchal evolution of endocrine pattern and ovarian aspects in adolescents with menstrual irregularities.

Ninety-five adolescents with menstrual irregularities persisting since menarche were studied and the data analyzed in relation to gynecologic age. In each year, in the premenstrual phase, luteinizing hormone (LH), testosterone (T), and androstenedione (delta 4A) values were higher than those of adults. Estrone (E1), estradiol (E2), 17-hydroxyprogesterone (17-OHP), progesterone (P), and dehydroepiandrosterone (DHEA) and its sulfate (DHEAS) gradually increased up to adult values correlating with gynecologic age. By ultrasound, nearly half the ovaries were multicystic and ovarian volume was greater than that of adults in each gynecologic year. When the data were classified as ovulatory or anovulatory, ovulation appeared to be a dynamic process characterized by ovaries similar to those of adults and by increasing hormonal levels correlated to gynecologic age. By contrast, in anovulatory cycles, mean T, delta 4A, and LH values were stable in each year and constantly higher than in ovulatory cycles and adult controls. These findings suggest that, despite persistent irregular cycles, some adolescents normalize all endocrine and ovarian parameters toward maturity, while subjects with persistent irregular anovulatory cycles maintain marked hyperandrogenism, increasingly high LH values, and enlarged multicystic ovaries.

EPISODIC PULSATILE SECRETION OF FSH, LH, PROLACTIN, OESTRADIOL, OESTRONE, AND LH CIRCADIAN VARIATIONS IN POLYCYSTIC OVARY SYNDROME

Pulsatile secretion of LH, FSH, PRL, oestradiol and oestrone was studied in a group of 16 patients with micropolycystic ovary syndrome (PCOS) and compared with that of normal ovulatory women in the fifth to sixth day of the cycle. Hormone concentrations were measured at 10 min intervals for 8 h starting at 0930 h. In seven subjects, the study was prolonged for 24 h, with 20 min interval samples, in an attempt to evaluate the circadian rhythm of LH by cosinor analysis. Significant fluctuations occurred in the concentration of each hormone. Values shown are mean ± SD. PCOS subjects had high LH mean values (27.9 ± 5.9 IU/1) (P< 0.005). LH pulse amplitude was higher than controls (11 6 ± 3.7 IU/1 versus 5.2 ± 1.8 IU/1; P<0.005) while no consistent changes in frequency or interpulse interval (62.0±10.7 min versus 65.8 ± 19.2 min; P= NS) were found. A mean of 4.8 ± 1.2 pulses of FSH occurred in 8 h and the mean pulse amplitude was 2.68 ± 1.11 with no differences from controls. All patients were normoprolactinaemic. A mean of 5.5 ± 1.9 pulses occurred in 8 h, the interpulse interval was 76.1 ± 14.4 min and the amplitude was 2.87 ± 0.76 ng/ml and there were no significant differences from controls; 75% of PRL pulses showed a temporal relationship with LH pulses. Oestrone mean basal values were higher in PCOS (47.2 ± 12.5 pg/ml) than controls (32.0 ± 9.9 pg/ml; P<002), while no differences were observed as regards oestradiol. Oestradiol pulse amplitude was nearly the same as oestrone (43.6 ± 18.8 pg/ml and 37.7 ± 16‐1 pg/ml, respectively); 6‐0 ± 2.2 pulses and 6.0 ± 1.6 pulses occurred in 8 h with an interpulse interval of 81.1 ± 27.1 min and 71.8 ± 11.1 min, respectively. Sixty‐five per cent of LH pulses were followed by an oestradiol and oestrone peak. The mean time of the appearance was 17 ± 15 min and 25 ± 23 min, respectively. In the PCOS group a consistent 24 h rhythm in mean plasma

Technical aspects of oocyte cryopreservation

Since the successful development in the mouse, the oocyte cryopreservation has been applied with varying success to a number of different species including the human. The recently reported successes in terms of pregnancies obtained by human oocyte cryopreservation are encouraging. Several studies typically reported different rates of survival (20-80%), fertilization (30-60%) and cleavage (32-100%). This variability of results throws some doubts on the usefulness of oocyte cryopreservation in IVF treatment cycles. It remains to be determined whether the relatively different success rates reported in literature, mainly in terms of survival rate, are due to methodological differences. We tried to investigate the effect of some factors on the oocyte survival rate after thawing: the presence or absence of cumulus oophorus and the exposure time of the oocytes to cryoprotectant. We suggest that a combination of several factors including both morphological and biophisical ones can affect the oocyte survival rate.

Human urinary follicle-stimulating hormone and human menopausal gonadotropin in induction of multiple follicle growth and ovulation

Five normally menstruating women were treated, in an attempt to induce development of multiple follicles, with pharmacologic doses of purified human urinary follicle-stimulating hormone (hU-FSH) and (in another instance) with human menopausal gonadotropin (hMG) administered on the second and third days after the onset of menses. All of the cycles were ovulatory: the follicular phase was short and the luteal phase length was normal in both hMG and hU-FSH treatment. No substantial differences were seen between the two types of treatment in regard to plasma values of FSH, luteinizing hormone (LH), estradiol (E2), testosterone, and progesterone (P). FSH, E2, and P increased to supraphysiologic levels, and LH fluctuated within the normal range. On ultrasound examination, a large number of growing and matured follicles were visualized during both treatments: at human chorionic gonadotropin administration, multiple preovulatory follicles (greater than or equal to 15 mm) and only a few small follicles (less than 10 mm) were imaged, without any difference between the two types of treatment. Multiple corpora lutea were often obtained. These data underline that pharmacologic doses of FSH alone are able to induce the growth of multiple preovulatory follicles when the initiation of stimulation is timed early. Besides this, exogenous LH does not seem to interfere with follicular recruitment, and it is not required for follicular maturation and ovarian steroidogenesis when endogenous normal LH mean values are present.

Induction of ovulation with human urinary follicle-stimulating hormone: endocrine pattern and ultrasound monitoring☆

Nine infertile patients, suffering from polycystic ovaries, were treated with human urinary FSH and hCG (eight cases) to induce ovulation. Oestrone, oestradiol, 17 alpha-hydroxyprogesterone, testosterone and androstenedione serum levels increased during the treatment. A decrease in luteinizing hormone serum levels was noticed, and in five cases a spontaneous peak was observed. No changes were noted in 5 alpha-dihydrotestosterone serum levels. Ultrasound scanning of the ovaries often revealed multiple follicles at different speeds and stages of growth and their marked turnover was observed. The beginning of the LH spontaneous surge was precocious compared to the normal ovulating follicular sizes: it does not appear that an optimum size exists but when the LH peak occurred too prematurely, ovulation did not take place. Administration of hCG seems to be necessary as spontaneous peaks of LH are not always followed by rupture of the follicle. Ultrasound scanning plays an important role in monitoring ovulation induction, while oestradiol and 17 alpha-hydroxyprogesterone serum levels are not good indicators of follicular maturity in multifollicular growth. Ovulation occurred in eight patients; four conceptions were obtained, one of which resulted in abortion. No ovarian hyperstimulations were observed.

Comparison between Human Urinary Follicle-Stimulating Hormone and Human Menopausal Gonadotropin Treatment in Polycystic Ovary

Five infertile patients with polycystic ovarian disease were treated to induce ovulation with pure human urinary follicle-stimulating hormone and human menopausal gonadotropin consisting of follicle-stimulating hormone and luteinizing hormone in 1:1 ratio. No substantial differences were seen between the two types of treatment regarding plasma values of follicle-stimulating hormone, prolactin, testosterone, dihydrotestosterone, progesterone, and 17-hydroxypro-gesterone. Estrone, estradiol, and androstenedione values were higher during human urinary follicle-stimulating hormone treatments. Luteinizing hormone levels dropped in both treatments, but the fall was greater during human urinary follicle-stimulating hormone. No real differences were observed concerning number of ovulations, length of treatments, and follicle-stimulating hormone amounts administered; no hyperstimulations were observed. These data do not confirm the observation that more controlled responses of the ovaries can be elicited when low luteinizing hormone gonadotropin preparations are used.