Vaginal gel versus intramuscular progesterone for luteal phase supplementation: a prospective randomized trial.

The aim of this randomized study was to compare the efficacy of intramuscular progesterone (IMP) and progesterone in vaginal gel (VGP) at two different doses for luteal support in IVF. A total of 412 patients, aged between 28 and 37 years, were randomized into three groups. The day after oocyte retrieval each patient began supplementation with one of the following: IMP 50 mg daily (150 patients), VGP 90 mg once daily (143 patients), or VPG 90 mg twice daily (148 patients). No significant difference was found between the three groups in any of the endpoints. The rate of positive beta-human chorionic gonadotrophin per transfer was 38.4% with IMP, 35.0% with VPG once daily and 43.1% with VPG twice daily. Clinical pregnancy rate per transfer and implantation rate were 32.6% and 19.6% with IMP, 26.3% and 16.4% with one dose of VGP, and 37.2% and 21.1% with two doses of VGP. Live birth rate per transfer was 26.1%, 23.4% and 29.9%, respectively. Progesterone vaginal gel can be successfully used as an alternative to intramuscular progesterone for luteal support in IVF. One daily dose appears sufficient to induce clinical pregnancies and live births at a rate comparable to intramuscular supplementation.

Clinical relevance of sperm DNA damage in assisted reproduction.

Many studies have shown how a 'paternal effect' can cause repeated assisted reproduction failures. In particular, with increasing experience of intracytoplasmic sperm injection (ICSI), it became evident that spermatozoa from some patients repeatedly fail to form viable embryos, although they can fertilize the oocyte and trigger early preimplantation development. Many authors have shown how this paternal effect can be traced back to anomalies in sperm chromatin organization: the spermatozoa of subfertile men are characterized by numerical abnormalities in spermatozoal chromosome content, Y chromosome microdeletions, alterations in the epigenetic regulation of paternal genome and non-specific DNA strand breaks. In particular, pathologically increased sperm DNA fragmentation is one of the main paternal-derived causes of repeated assisted reproduction failures in the ICSI era. The intention of this review is to describe nuclear sperm DNA damage, with emphasis on its clinical significance and its relationship with male infertility. Assessment of sperm DNA damage appears to be a potential tool for evaluating semen samples prior to their use in assisted reproduction, helping to select spermatozoa with intact DNA or with the least amount of DNA damage for use in assisted conception.

Cumulative pregnancy rates resulting from the use of fresh and frozen oocytes: 7 years' experience.

Storing supernumerary embryos and transferring them later fully utilizes the reproductive potential of retrieved oocytes, allowing a significant increase in the overall number of pregnancies achieved from a single cycle of ovarian stimulation treatment. As an alternative to embryo cryopreservation, preservation of unfertilized oocytes has been proposed to maximize clinical outcome. This paper presents data concerning the cumulative pregnancy rate after use of fresh and cryopreserved oocytes. In 80 treatment cycles in which patients chose to have only a few fresh oocytes inseminated, 24 pregnancies were obtained (30.0%), with an implantation rate of 22.6%. After cryopreservation with the standard slow-cooling protocol, the survival, fertilization and cleavage rates of 918 frozen oocytes were 43.4, 51.5 and 86.0% respectively. A total of 14 frozen pregnancies were achieved, with pregnancy rate 19.2% per transfer and implantation rate 12.3%. The cumulative pregnancy rate was 47.5% per patient. Therefore, despite a low rate of oocyte post-thaw survival, it appears that oocyte storage appreciably improves the number of pregnancies per treatment cycle in cases in which only a minority of oocytes are destined for the fresh treatment. This outcome provides valuable information for appraising the chances of clinical success when the option of embryo cryopreservation is not available.

Live birth after IVF in a 46-year-old woman.

Pregnancies after IVF have been reported in women aged > or =44 years, but nobody older than 45 years at oocyte retrieval delivered. We report a case of birth of a healthy child after IVF in a 46-year-old infertile woman. Ovarian stimulation was performed with clomiphene citrate, 150 mg daily for 5 days. Three oocytes were retrieved and one embryo was replaced. The patient delivered a healthy male infant after Caesarean section at 39 weeks. A successful pregnancy after IVF with homologous oocytes can be achieved in women older than 45 years. At this age IVF is not a cost-effective treatment compared with oocyte donation, but it may be offered in countries in which gamete donation is forbidden.

Implantation markers and endometriosis.

The receptive phase of the endometrium seems to occur in close association with the appearance of pinopodes and endometrial integrins that may be activated by the interleukin-1 system (IL-1). Embryo attachment is the result of adhesion protein expression, and the invasion of the embryo is governed by proteolytic enzymes. Leukaemia inhibitory factor (LIF) is produced by natural killer lymphocytes that interact with the invading trophoblast. This may activate urokinase plasminogen activator (uPA) and gelatinase enzymes, which play a crucial role in trophoblast invasion. Oestrogen stimulates, while progesterone inhibits, LIF. The role of endometrial contractility in displacing human embryos from the Fallopian tube to the lumen cavity of the uterus or vagina in terms of pregnancy or wastage is still a matter of discussion. Endometriosis and its associated abnormal uterine contractions may be also linked to implantation failure. Unless new evidence emerges to indicate otherwise, it can be assumed that progesterone is, either in a direct (non-genomic: contractility) or indirect (genomic: decidualization) manner, the only determinant of endometrial priming necessary for embryo nidation.

Predictive factors for embryo implantation potential.

In spite of recent improvements in IVF, pregnancy rates have not increased significantly and one of the major problems remains the high multiple pregnancy rate. Better criteria are therefore necessary to establish the viability of a transferable embryo. Early prognosis of the developmental fate of the oocyte would help in selecting the best embryos to transfer, but non-invasive selection at the oocyte stage (extracytoplasmic and intracytoplasmic morphology) has proved to be of little prognostic value. Recently, it has been shown that follicular vascularization appears to be predictive of oocyte developmental fate, making it a good first-step approach for selection. Observation of pronuclei patterns at the zygote stage appears to offer an additional prognostic tool, correlating well with IVF outcome. Morphological evaluation of the embryo at days 2-3 remains the most used and valid method of selection, even though it is not sufficient to select embryos with the higher implantation potential. Blastocyst culture is another possible strategy for selecting the best embryos with reduced risk of aneuploidies, though not all major chromosomal aberrations are excluded by prolonged in-vitro culture. In summary, selecting the best embryo for transfer is a decision that should be based on choices made during the different stages of assisted reproductive technologies.

Pregnancies and births after oocyte cryopreservation.

OBJECTIVE: To determine the potential of oocyte cryopreservation techniques. DESIGN: Retrospective data analysis. SETTING: A tertiary infertility center. PATIENT(S): Sixty-eight patients (29 to 37 years of age) undergoing assisted reproduction procedures for infertility problems. INTERVENTION(S): Oocytes from women treated for infertility were cryopreserved with a slow cooling/rapid thawing protocol in which 1,2 propanediol and sucrose were used as cryoprotectants. Eighty-six thawing cycles were performed. MAIN OUTCOME MEASURE(S): Rates of survival after thawing, fertilization after intracytoplasmic sperm injection, cleavage, implantation, and pregnancy. RESULT(S): We treated 68 patients through 86 thawing cycles. Seven hundred thirty-seven oocytes were thawed, and 59 transfer cycles were performed. The survival rate was 37%. The fertilization and cleavage rates were 45.4% and 86.3%, respectively. A total of 15 clinical pregnancies were achieved with pregnancy rates of 25.4% per transfer and 22% per patient. There were three miscarriages, resulting in an abortion rate of 20%. Seventeen of the 104 transferred embryos implanted, corresponding to an implantation rate of 16.4%. Thirteen babies were born, 8 females and 5 males. CONCLUSION(S): Statistically significant results were obtained for fertilization, cleavage, and pregnancy rates. Our results show oocyte cryopreservation may represent an alternative to embryo storage in selected cases.

Perifollicular vascularity monitoring and scoring: a clinical tool for selecting the best oocyte.

The aim of our study was to assess relationship between perifollicular vascularity and outcome of in vitro fertilization (IVF) cycles. In our study, women who received embryos originating from oocytes developed in well vascularized follicles had a statistically higher pregnancy rate than women who received embryos chosen without considering follicles of origin.

The patterns of uterine contractility in normal menstruating women: from physiology to pathology.

The nonpregnant uterus shows uterine activity throughout the menstrual cycle. This uterine activity was detected both by single strep tissue and by using intrauterine pressure recordings in vitro and in vivo. Today, ultrasound has made it possible to study this activity with a noninvasive approach and to assess uterine contractions (UCs) in real-time movements of the uterus. Throughout the menstrual cycle, wavelike activity patterns of the reproductive organ were established. These patterns are under control of steroid hormones. Adequate UCs may provide for gamete/embryo transportation throughout the uterotubal cavities and successful embryo implantation in spontaneous and/or assisted reproduction. Inadequate UCs may produce ectopic pregnancy, miscarriages, retrograde bleeding with dysmenorrhea, and endometriosis.

Psychological distress of conservative and nonconservative uterine surgery: a prospective study.

OBJECTIVE: The aim of the present study is to evaluate the psychological reaction to conization before and after the operation compared to hysterectomy. To study the incidence of psychological stress related to conization, 60 women undergoing conization were compared to 40 women who had undergone hysterectomy. METHOD: Psychological disease was rated in the pre- and postoperative periods using the Symptom Questionnaire (SQ). Each patient was evaluated 2 weeks before the operation, and 3, 6 and 12 months after it. RESULTS: Both the conization and hysterectomy groups showed a significant reduction in anxiety and depression at the 3-, 6- and 12-month follow-ups compared to the preoperative period. Two weeks before surgery, 8 conization patients (19.5%) showed an anxious status, while 10 (24.3%) presented high levels of anxiety and depression. Within hysterectomy patients, the occurrence was respectively of 4 (12.9%) and 10 women (32.2%). Twelve months after surgery, of the women with preoperative depression, only four (9.7%) conization and four (12.9%) hysterectomy patients presented a negative mood status. A similar trend was present for somatic symptoms but only in the conization group, because the hysterectomy patients did not show a reduction in these symptoms from the preoperative to the postoperative period. This result could be related to the surgical menopause due to the bilateral oophoriectomy executed in more than half of the hysterectomy group. CONCLUSION: In general, the results of the present study show that the conservative and nonconservative uterine surgery determines a good psychological prognosis in the short- and long-term postoperative periods.

Isolated premature pubarche: ultrasonographic and color Doppler analysis--a longitudinal study.

Twenty-seven girls with premature pubarche were studied by ultrasonographic and color Doppler analyses to determine the incidence of polycystic ovaries (PCO), to longitudinally assess their evolution, and to search for any hormonal correlation. The girls were submitted to auxological, clinical, and hormonal evaluation, and 21-hydroxylase deficiency was ruled out by an ACTH test. Furthermore, the girls underwent ultrasonographic and color Doppler ovarian and uterine analyses. Among girls with premature pubarche, the prevalence of PCO was 41%. Advanced skeletal maturation, tall stature, and increased hair distribution were constant in these patients. In patients with ultrasonographic and color Doppler evidence of PCO, the ovarian volume, the number of small-sized subcapsular follicles, the stromal echogenicity, and the ovarian stromal vascularization progressively increased during the study. In the whole studied population, ovarian volume correlated with the number of small-sized follicles (r = 0.719; P < 0.0001). Furthermore, a slight and inverse correlation has been found between ovarian volume and ovarian stromal artery pulsatility index (r = -536; P = 0.048). In conclusion, we affirm that PCO are greatly represented among girls with premature pubarche and progressively evolve.

Characteristics of uterine contractility during menses in women with mild to moderate endometriosis.

OBJECTIVE: To establish the role of uterine contractions in retrograde menstruation with subsequent abdominal implantation of endometrial tissue. DESIGN: Controlled prospective study. SETTING: University hospital-based study. PATIENT(S): Infertile women with (n = 22) and without (n = 22) endometriosis. MAIN OUTCOME MEASURE(S): Frequency, amplitude, and basal pressure tone of uterine contractions; correlation of contractions with retrograde bleeding and presence of viable endometrial cells; and dysmenorrhea before and 3 and 24 months after surgery. RESULT(S): Compared with controls, patients with endometriosis had uterine contractions with higher frequency (22.73 +/- 5.66 osc/10 min vs. 11.09 +/- 3.26 osc/10 min), amplitude (20.83 +/- 3.94 mm Hg vs. 6.77 +/- 2.83 mm Hg), and basal pressure tone (50.14 +/- 16.30 mm Hg vs. 24.68 +/- 6.14 mm Hg). Dysmenorrhea was scored as 4.09 +/- 1.44 in patients with endometriosis and 0.86 +/- 1.42 in controls. Retrograde bleeding was found in 73% of patients with endometriosis vs. 9% of controls, and only 45% of patients with endometriosis had viable endometrial cells in the cul-de-sac. CONCLUSION(S): Endometriosis may result from abnormal myometrial contractility through tubal transportation, dissemination, and implantation of endometrial viable cells into the abdomen.

Twenty-one day administration of dienogest reversibly suppresses gonadotropins and testosterone in normal men.

Androgen-progestin combinations are promising male contraceptive regimens. Optimization of these regimens includes the development of new progestins with more favorable biological properties. In this context we tested the effects of the progestin dienogest (DNG) on reproductive hormones and metabolic parameters in men. After a 3-wk control period, 25 men were randomly assigned to receive daily doses of 2, 5, or 10 mg DNG or placebo and 10 mg cyproterone acetate for 21 d (n = 5 subjects/group). Subjects were followed for 2 wk after cessation of hormone administration. Weekly blood samples, physical examinations, hormone and chemistry measurements, semen analysis, and sexual/behavioral assessments were performed. These parameters were compared among study groups and within each group at different time points throughout the study periods. DNG and cyproterone acetate administration resulted in profound suppression of gonadotropins and T with no change in SHBG levels. No adverse effects were detected in any subject. Hormone levels returned to baseline after stopping hormone intake. DNG is a potent suppressor of gonadotropins and T in men and does not induce adverse effects over a 21-d administration. DNG is a promising progestin to be used in clinical trials for male contraception.

Endometrial preparation for frozen-thawed embryo transfer with or without pretreatment with gonadotropin-releasing hormone agonist.

OBJECTIVE: To test the efficacy of endometrial preparation with exogenous steroids, without pretreatment with gonadotropin-releasing hormone (GnRH) agonist, in women with normal ovarian function. DESIGN: Prospective randomized study. SETTING: Private outpatient infertility clinic. PATIENT(S): Two hundred ninety-six women undergoing frozen-thawed embryo transfer. INTERVENTION(S): In group 1 (146 patients), depot GnRH agonist was administered in the luteal phase; treatment with 17beta-estradiol transdermal patches at steadily increasing dosage from 100 to 300 microg was then given for at least 12 days. In group 2 (150 patients), endometrial preparation began on day 1 of menstrual cycle. The starting dose was 200 microg; this was increased to 300 microg after 7 days. MAIN OUTCOME MEASURE(S): Pregnancy, abortion, implantation and cancellation rates. RESULT(S): In group 2, six cycles (4%) were cancelled due to evidence of ovulation. Groups were similar in the percentage of embryos that survived freezing-thawing (77.1% in group 1 and 76.6% in group 2) and in the number of embryos transferred per patient (2.1 +/- 0.6 and 2.1 +/- 0.7, respectively). Groups 1 and 2 did not differ significantly in rates of pregnancy (19.7% and 24.1%), abortion (17.8% and 11.7%), and implantation (10.4% and 11.9%). CONCLUSION(S): Endometrial preparation for frozen-thawed embryo transfer based exclusively on steroid administration appears to be as effective as the more conventional protocol involving preliminary desensitization with a GnRH agonist. This simplified protocol reduces costs, minimizes pharmacologic treatment, and increases patient compliance.

Adjuvant L-arginine treatment in controlled ovarian hyperstimulation: a double-blind, randomized study.

BACKGROUND: Enhanced vascularization appears to be important for follicular selection and maturation in both spontaneous and stimulated IVF cycles. Nitric oxide, formed in vivo from L-arginine, may play a key role in follicular maturation and ovulation. METHODS: To evaluate the role of L-arginine supplementation in controlled ovarian hyperstimulation, 37 IVF patients were divided into two groups according to ovarian stimulation protocols: group I, GnRH agonist plus pure (p)FSH plus oral L-arginine (n = 18); and group II, GnRH agonist plus pFSH plus placebo (n = 19). Hormonal, ultrasonographic and Doppler evaluations were performed, and plasma and follicular fluid nitrite/nitrate concentrations were monitored. RESULTS: Thirty-two patients completed the study. In group I (n = 16), plasma L-arginine concentrations increased from (basal) 87 +/- 12 micromol to 279 +/- 31 micromol (P = 0.002) on the day of beta-HCG administration. In this group, pFSH treatment was shorter (P = 0.039) than in group II (n = 16). The number of the follicles > or =17mm was lower (P = 0.038) in group I than group II. The "good quality" embryos were fewer in number (P = 0.034) and pregnancy rate, both per patient (P = 0.024) and per embryo transfer (P = 0.019), was lower in group I. In the L-arginine group, an increased follicular fluid concentration of nitrite/nitrate was observed. On day 8 of the cycle, elevated plasma estradiol levels were associated with decreased blood flow resistances of perifollicular arteries. Follicular fluid concentrations of nitrite/nitrate were inversely correlated with embryo quality (r = -0.613; P = 0.005) and perifollicular artery pulsatility index (r = -0.609; P = 0.021). CONCLUSIONS: L-Arginine supplementation may be detrimental to embryo quality and pregnancy rate during controlled ovarian hyperstimulation cycles.

Polycystic ovaries in childhood: a common finding in daughters of PCOS patients. A pilot study.

BACKGROUND: Polycystic ovarian syndrome (PCOS) is a controversial endocrine pathology and, recently, it has been suggested that the condition is hereditary. The aim of this study was to prospectively determine in daughters of patients with PCOS, by ultrasonographic (US) and colour Doppler analyses, the incidence of polycystic ovaries and search any correlation with specific hormonal parameters. METHODS: Fifteen prepubertal offspring (Group I) of patients with PCOS and 10 normal control prepubertal girls (Group II) were submitted to clinical, auxological, and basal hormonal assay. In addition all patients were submitted to US and colour Doppler ovarian and uterine evaluation. RESULTS: Among Group I girls the prevalence of polycystic ovaries was 93%, whereas no subjects among Group II had polycystic ovaries. The ovarian volume (2.76 +/- 1.21 ml versus 0.87 +/- 0.46 ml; P < 0.001) and the number of small sized follicles (5.36 +/- 2.2 versus 0.75 +/- 0.92; P < 0.001) were significantly higher in Group I than Group II patients. In addition, a normal stromal score and an absent stromal vascularization was observed in the control group. The hormone levels did not differ between the two groups. CONCLUSION: In conclusion we speculate that polycystic ovaries in childhood may be considered a sign of genetic predisposition to PCOS and that environmental factors may express the adult clinical and hormonal presentation of the syndrome.