Urinary LH Research Articles

Pituitary-Gonadal Function in Adolescent Males Born Appropriate or Small for Gestational Age with or without Intrauterine Growth Restriction

Being born small for gestational age (SGA) is suggested to influence female pituitary-gonadal axis, but only a few studies have focused on male pituitary-gonadal function. Our objective was to evaluate the influence of fetal growth rate on male reproductive function. We conducted a follow-up study of a prospective study with data on third trimester fetal growth velocity and birth weight. The study was conducted at Copenhagen University Hospital. Fifty-two healthy adolescent males participated. They were divided into those born appropriate for gestational age (AGA) and SGA, with or without intrauterine growth restriction. Pubertal stage, testicular size, and reproductive hormones were determined. Overnight 20-min LH and FSH profiles and overnight urine LH and FSH were determined in an additional study (n=30). No significant differences were found in testosterone levels (19.2 vs. 18.9 nmol/liter), inhibin B levels (186.5 vs. 188.0 pg/ml), or LH/testosterone ratio (0.15 vs. 0.18) between AGA and SGA, respectively. No significant differences in overnight secretory patterns of gonadotropins or testicular size and morphology were determined by ultrasonography between AGA and SGA. Fetal growth velocity did not influence any of the reproductive hormone levels. Overnight urinary LH and FSH excretion correlated statistically significantly with overnight LH (r=0.50; P=0.02) and FSH (r=0.44; P=0.04) secretion, respectively. Poor third trimester growth and/or low birth weight had no effect on subsequent male reproductive hormones. Contrasting a previous study, we found no difference in testosterone or inhibin B levels between SGA and AGA, suggesting that testicular function was not impaired in adolescent males born after compromised fetal growth.

Human chorionic gonadotropin administration vs. luteinizing monitoring for intrauterine insemination timing, after administration of clomiphene citrate: a meta-analysis

To systematically compare hCG administration as a method for intrauterine insemination (IUI) timing with conservative urinary LH surge detection in infertility treatment. Meta-analysis of prospective and retrospective trials. Tertiary fertility and IVF center. One thousand four hundred sixty-one patients who received hCG after a clomiphene citrate regimen, compared with 1,162 patients who had an LH surge detection for IUI timing. Both MEDLINE and Cochrane Collaboration were searched. References of retrieved articles were included in the search. The meta-analysis included all controlled trials examining the effectiveness of hCG administration before IUI on clinical-pregnancy rates in comparison with LH detection. Two independent reviewers performed data extraction. Pregnancy rates. Seven studies with 2,623 patients were included in the meta-analysis (1,461 patients received hCG, and 1,162 had LH surge detection). When all studies were combined, patients who received hCG before IUI demonstrated lower clinical-pregnancy rates than did women who had IUI after spontaneous ovulation (odds ratio, 0.74; 95% confidence interval, 0.57-0.961). In subgroup analysis of studies that considered ovulatory dysfunction to be the infertility reason, the results favored women who received hCG. In contrast, across studies that reported male factor as the infertility reason, as well as across studies including women with unexplained infertility, results appeared to favor the LH surge detection approach. However, none of those subgroup analyses reached statistical significance. Available data do not demonstrate a consistent, clinically important benefit of hCG-induced ovulation compared with spontaneous ovulation for IUI timing.

P-780: Impact of the use of recombinant LH (RLH) versus urinary LH (uLH) in conjunction with recombinant FSH (rFSH) in GnRH agonist and antagonist-treated IVF cycles

Controversy exists as to the indication, dosage and type of preparation when considering LH addition to rFSH-treated IVF cycles. We aimed to compare the impact of the use of rLH versus uLH in conjunction with rFSH in IVF cycles treated with a GnRH agonist or antagonist in patients with anticipated low response.

P-1000: Evaluation of pregnancy rates following intrauterine inseminations using a commercially available sperm preparation kit (Sperm Select® System)

To compare pregnancy rates following IUI between the Sperm Select® kit and standard sperm preparation techniques. Retrospective review. This study included 3245 non-donor IUI in clomiphene or hMG stimulated cycles. IUI was timed 24-36 hours after positive urine LH surge or intramuscular administration of 10,000 units of hCG. Sperm Select® System preparations were performed exclusively at one of our satellite clinics according to the manufacturer’s directions. Standard sperm preparations were performed in a dedicated andrology laboratory and included straight wash, density gradient or swim-up techniques. Pregnancies were determined after at least two rising serum hCG levels. Pregnancy frequencies were compared using Chi-square statistical analysis. Pregnancy rates following IUI were similar for standard versus Sperm Select® System preparations (p=0.83, Table 1). Furthermore, there was no statistically significant difference between Sperm Select® System and any of the individual standard sperm preparation techniques (Table 2). The pregnancy rates varied with the type of ovarian stimulation. However, there was no statistically significant difference based on the type of sperm preparation used. Clomiphene cycle pregnancies were 10.31% with the Sperm Select® System and 9.49% with the standard preparation. HMG cycle pregnancies were 12.35% with the Sperm Select® System and 13.60% with the standard preparation.Tabled 1Tabled 1 The Sperm Select® System yields pregnancy rates after IUI comparable to standard sperm preparation techniques. It is a viable alternative for clinics without access to a specialized andrology laboratory.

L-Selectin Ligand in Endometrium From Women With Infertility

To evaluate the pattern and intensity of the L-Selectin ligand in the endometrium from both fertile and infertile women throughout the menstrual cycle and during the window of implantation. This is a retrospective study on 48 endometrial samples from reproductive-aged women at various times in the menstrual cycle. Immunohistochemistry and western blot analysis was performed using the antibody, MECA-79, which recognizes the ligand for the L-selectin adhesion molecule. Endometrial biopsies were obtained from both fertile and infertile women during the menstrual cycle. Secretory phase samples were timed to the urinary LH surge. Overall, 48 samples were compared. Eighteen biopsies from normal cycling women included 3 proliferative, 3 early secretory (day15-19), 10 mid-secretory (day 20 to 24) and 2 late secretory phase (day 25-28). Additionally, mid-secretory samples were obtained from 7 women with PCOS in ovulatory cycles and 17 women with endometriosis, and 6 samples with significant histologic delay and luteal phase defect (LPD). These were compared to biopsies from normal fertile women in the mid-secretory phase. Statistical analysis was performed using ANOVA with Bonferroni correction with significance assigned at the 95% confidence interval (p < 0.05). The L-selectin ligand was absent during the proliferative phase and peaks during the early and mid-secretory phase in both glands (p=0.002 and 0.003) and lumen (p=0.004 and 0.005). In this first comparison of the L-selectin ligand in infertile and fertile women, we show that there was a slight decrease in overall expression of this antigen in glandular epithelium of women with PCOS and endometriosis (P =0.07 and P=0.052, respectively), compared to normal controls. However, in a subset of these patients the loss of L-selectin ligand was greater. A significant decrease was seen in the luminal expression in both PCOS and endometriosis patients (p=0.02 and p=0.048, respectively). Endometrium from women with LPD also had significantly less MECA79 staining on both glands (p=0.02) and lumen (p = 0.03), respectively. These findings were corroborated using western blot analysis. Given the suspected role in L-selectin as the attachment receptor for the human embryo, these results shed light on the maternal-fetal interface during the implantation window. The identification of depressed expression of luminal and glandular L-selectin ligand in a subset of women with PCOS and endometriosis provides further evidence for selective defects in endometrial receptivity in certain groups of women with infertility. The L-Selectin ligand is significantly reduced in some women with PCOS and endometriosis at the time of implantation. Further studies will be required to investigate the mechanism for this decrease, but evidence points to a reduction in the N-acetylglucosamine-6-O-sulfotransferase activity. Screening for such defects may be important prior to IVF or other infertility treatments.

The use of urine LH detection kits to time intrauterine insemination with donor sperm

The study was carried out to determine the most likely time of day for the onset of the LH surge as detected using urine LH dipsticks, and to calculate the optimum time interval from the onset of the LH surge to intrauterine insemination (IUI). A prospective study of 1540 cycles of IUI with donor sperm at Cleveland Fertility Centre, Middlesbrough, between June 1990 and February 2004. Only 951 cycles (where a positive urine LH dipstick result was immediately preceded by a negative result) were included in our study. To determine the best time interval between the onset of the LH surge and IUI, women were divided into five subgroups according to the positive urine test-IUI time interval and the pregnancy rate and live birth rate per cycle were calculated for each group. The first positive test was most frequently (44.5%) found at lunch-time (11:00-15:00). The live birth per cycle achieved was 5.6% when the insemination was performed 18-23 h from the first detection of the LH surge, and 11.7% when it was performed between 24 and 42 h. The live birth rate declined to 6.5% when IUI was performed later than that. Overall, no significant differences were discovered in live birth or pregnancy rate when insemination was performed at any of the time points between 18 and 53 h. Our study suggested that lunch-time is the best time to check for the LH surge using urine dipsticks and insemination at any time between 18 and 53 h after the onset of the surge will produce optimal results.

Charles Thibault and assisted reproduction in France

Charles Thibault was liked by French gynaecologists. There was not a year that Charles Thibault did not attend clinician gynaecology conferences. He made great strides in research on in vitro fertilisation, being the first to perform in vitro fertilised (IVF) oocyte transfers in rabbits. Later, in 1978 the first human pregnancy following IVF was achieved in the UK when Louise Brown was born. In 1980, two French teams,one at the Sèvres hospital and the other at the Clamart University Teaching Hospital, carried out egg retrievals in patients with natural cycles, after determination of the urinary LH peak, under general anaesthesia and by laparoscopy. The Clamart team developed LH SIR, which enabled a more accurate determination of the ideal time for egg collection. In 1983, the same team reported the first ambulatory oocyte retrievals by ultrasound, under local anaesthesia. This new technique did not require general anaesthesia. Finally, in 1983, the rate of births, per transfer, for the Sèvres team rose to 5.31%. 1984 showed considerable improvement: 13.83%. The first step in establishing IVF in France was completed with the Carghese symposium, in September 1984, where Charles Thibault pleaded for animal experimentation before human clinical trials. It was only later that ART developed significantly, necessitating a legislative framework and organisations such as GEFF and FIVNAT.

Clinical study of ion-intreoduction therapy in patients with dysfunction ovulation

目的 探讨中药内服外用治疗卵巢排卵功能障碍所致不孕症的临床疗效.方法选择卵巢排卵功能障碍所致不孕患者45例,治疗组采用中药内服配合离子导入治疗,对照组仅内服中药,观察两组治疗前后的基础体温曲线(BBT)、尿液黄体生成激素(LH)等变化.结果中药内服外治诱发排卵的总有效率为86.6%,治疗组疗效明显高于对照组(P<0.01).结论中药内服配合离子导入治疗排卵功能障碍有较好的临床疗效。

Rapid Resumption of Ovulation and Menstruation Following Continuous Use of the Combination Oral Contraceptive

Background: The return of ovulation and fertility following cyclic oral contraceptive (OC) use can be delayed in some women. The return to ovulation following long-term suppression with continuous use of the pill has not been studied. It is possible cycle suppression for many years could induce ovarian quiescence and a delay in the return to ovulation and menstruation. Objective: The purpose of this study was to evaluate the time to ovulation and menstruation following discontinuation of daily OC consumption. Materials and Methods: Three faculty, with continuous combination pill use for the prior three years or greater, prospectively stopped their pill use and underwent serial ultrasound examination, urinary LH testing to confirm ovulation, and serial serum estradiol and progesterone measurements. Result(s): Case 1 was a 42-year-old with 2 term deliveries who then took 20mcg ethinyl estradiol/100mcg levonorgestrel daily for 8 years. She ovulated 13 days following pill cessation but had an abnormally thin endometrium during the first month and bleeding did not begin until day 26. Case 2 was a 31-year-old nulligravid who took 35mcg ethinyl estradiol/1000mcg norethindrone daily for 5 and a half years. She had withdraw bleeding day 3, ovulated day 25 and had a menstrual bleed on day 33. Case 3 was a 31-year-old nulligravid who took 35mcg ethinyl estradiol/250mcg norgestimate daily for 3 years. She ovulated day 16 and had menses on day 30. Conclusion(s): There was no evidence of a significant delay in the return to ovulation or menses following years of cycle suppression and amenorrhea from continuous OC use in this small prospective study. None of these women were attempting to conceive, but it is possible conception could have occurred in the first cycle after cessation of daily OC consumption.

Interpretation of the BBT chart: using the “Gap” technique compared to the Coverline technique

Objective The aim of the study was to compare the accuracy of the Gap and Coverline techniques of interpreting the basal body temperature chart. Methods We compared the proportion of menstrual cycles for which the Gap and Coverline techniques accurately identified the post-ovulatory final fertile phase (FFP) and the initial infertile phase (IIP) and the median number of days each overestimated the fertile period, using urinary LH testing as the gold standard. Results The Gap and Coverline techniques identified the FFP within ±1 day of that identified by LH testing in 13/33 (39%) and 10/33 (30%) cycles ( χ 2=0.6; p=.44), respectively, and the IIP within +1 day of that of LH testing in 13/33 (55%) and 4/33 (12%) cycles, respectively ( χ 2=13.4; p<.001). The Gap and Coverline techniques overestimated the fertile period by 1 and 4 days, respectively (p=.0002). Conclusion Based on this small study, the Gap technique appears to be more accurate than the Coverline technique in identifying the post-ovulatory IIP.

Prevention of Endometrial Apoptosis: Randomized Prospective Comparison of Human Chorionic GonadotropinVersusProgesterone Treatment in the Luteal Phase

To study control of apoptosis in human endometrium, we examined late luteal-phase endometrial biopsies obtained in the late luteal phase for evidence of apoptosis and compared the effects of exogenous human chorionic gonadotropin (hCG) and progesterone on this process. Using a controlled, prospective, and randomized study design, 12 healthy, fertile, reproductive-age women (ages 20-34 yr) with regular menstrual cycles (range, 26-32 d) were recruited. Each underwent an endometrial biopsy 12 d after a urinary LH surge in a control and treatment cycle. After biopsy in a natural cycle, subjects were randomized to receive luteal doses of either 200 mg intravaginal progesterone (d 18-27) or a single im injection of 10,000 IU of hCG (d 19) followed by repeat endometrial biopsy and collection of serum on d 26. Apoptosis was assessed by DNA laddering, localizing apoptotic bodies using immunofluorescent labeling of DNA fragments (the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method), and immunohistochemical assessment of apoptosis markers bcl-2, bcl-x, and bax. Serum progesterone levels were compared between treatment groups. Evidence of apoptosis in control cycles was significantly reduced in endometrium after both luteal-phase treatments. The terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling results demonstrated significantly less apoptosis in the hCG treatment group compared with controls. Immunostaining for bcl-2 was higher in hCG- and progesterone-treated cycles, whereas bax expression was decreased and bcl-x immunostaining was not different between treatments. Serum progesterone levels were highest in the hCG-treated group, although statistical significance was not reached (P = 0.08). These results demonstrate that signs of apoptosis, already apparent by d 26 of the menstrual cycle can be reduced with either hCG or progesterone treatment. The clinical utility of these findings includes a rational use of luteal-phase support for treatment of women with infertility and/or recurrent pregnancy loss.

326 OPTIMIZING FREQUENCY OF FSH APPLICATION FOR SUPEROVULATORY TREATMENT IN CATTLE

The aim of the study was to examine the influence of the application frequency of Pluset® (Calier, Spain) and Folltropin® (Vetrepharm, London, Ontario, Canada) on the results of embryo recovery in Simmental cattle. Moreover, pulsatility of LH concentrations in plasma should be studied before and after application of Pluset. FSH was given in a treatment of eight or four injections on four consecutive days starting between Days 9 and 13 of the estrous cycle. Luteolysis was induced by application of 500 μg Estrumate® (Essex Tierarznei, Munich, Germany), applied 72 and 84 h after the first FSH injection. Embryo recovery was performed non-surgically on Day 7 of the following estrous cycle. Heifers received either a total amount of 500 IU Pluset according to the International Standard for human urine FSH and LH (P1: 8 injections, n = 20; P2: 4 injections, n = 54) or 200 mg Folltropin according to NIH bFSH-B1 (F1: 8 injections, n = 117; F2: 4 injections, n = 60). Cows received either 550 IU Pluset (P3: 8 injections, n = 54; P4: 4 injections, n = 183) or 260 mg Folltropin (F3: 8 injections, n = 36; F4: 4 injections, n = 159). Altogether 8 heifers were bled throughout 8 h every 10 min on Days 9 and 12 (before and during stimulation with FSH). The samples were analyzed for LH by ECLIA (intra-assay coefficient of variation (VK) 6.4%, inter-assay VK 8.9%). The evaluation of the data was carried out with the procedure ‘GLM’ of the statistics software package SAS (SAS Institute, Inc., Cary, NC, USA) and PULSAR analysis. As a post hoc test Student's t-test was used. Significance was set at P = 0.05. Results of embryo recovery are shown in the table. Number of LH pulses in 8 h (LSM ± SE) was 2.3 ± 0.4 on Day 9 and 0.6 ± 0.4 on Day 12 (P &lt; 0.05). We conclude that four injections of both FSH products can lead to results comparable with those reached with eight injections in a 12 hours interval. Therefore, the effort for superovulatory treatments can be reduced. However, it has to be considered that the portion of transferable embryos was reduced, partly significantly, after four FSH injections. This result was independent of the drug used. Table 1. Results of embryo recovery

Endometrial biopsy: A test whose time has come and gone

To document ovulation in an infertility investigation, a timed endometrial biopsy is painful, expensive, invasive, requires precise cycle timing for proper interpretation, and cannot independently distinguish between fertile and infertile women. Urinary LH kit testing has a significant false-positive rate (approximately 7.6%). A midluteal serum progesterone is a much simpler method that compares favorably with either endometrial biopsy or LH kit testing.

Histological dating of timed endometrial biopsy tissue is not related to fertility status

To assess the ability of histological dating to discriminate between women of fertile and infertile couples. The utility of histological dating of endometrium in the evaluation of infertile couples is uncertain. Prospective multicenter study, with subjects randomly assigned to biopsy timing. Criterion standard for infertility was 12 months of unprotected, regular intercourse without conception and for fertility at least one live birth within 2 years. University-based infertility practices. Volunteer subjects (847) recruited at 12 clinical sites participating in the National Institutes of Health-funded Reproductive Medicine Network. Inclusion criteria included ages 20-39 years, regular menstrual cycles, and no hormonal treatment or contraceptive use for 1 month before the study. Fertile controls were excluded if they had a history of infertility, recurrent pregnancy loss, or recent breastfeeding. Subjects underwent daily urinary LH testing. After detection of the LH surge, subjects were randomized to biopsy in the mid (days 21-22) or the late (days 26-27) luteal phase. Pathologists at each site estimated the cycle day based on standard criteria. For the primary analysis, an out-of-phase biopsy was defined as a greater than 2-day delay in the histological maturation of the endometrium. The proportion of out-of-phase biopsies in fertile and infertile women was compared using logistic regression models with age at randomization as a covariate. Comparisons were also made between fertile vs. infertile at the midluteal or late luteal phase time points. Biopsies were evaluated (301 mid and 318 late; N = 619). Out-of-phase biopsy results poorly discriminated between women from fertile and infertile couples in either the midluteal (fertile: 49.4%, infertile: 43.2%) or late luteal phase (fertile: 35.3%, infertile 23.0%). Results did not substantially differ using alternative definitions of "out-of-phase" or standardized cycle day. Histological dating of the endometrium does not discriminate between women of fertile and infertile couples and should not be used in the routine evaluation of infertility.

Absence of secretory endometrium after false-positive home urine luteinizing hormone testing

To examine the proportion of cases with proliferative endometrium on biopsies performed after positive home urine LH testing. Multicenter clinical trial of the usefulness of endometrial biopsy in the evaluation of infertility, with women from fertile and infertile couples randomly assigned to midluteal vs. late luteal phase endometrial sampling. Twelve clinical sites of the National Institutes of Health/National Institute of Child Health and Human Development-sponsored Reproductive Medicine Network. All women in the study had regular menstrual cycles. Fertile volunteers who had delivered a live born infant within the past 2 years without medical intervention were recruited through advertisements at participating sites. Infertile women with regular cycles were recruited from the clinical practices of the sites' physicians. Interview, informed consent, subject-interpreted home urine LH testing, and endometrial biopsy in either the midluteal or late luteal phase. Proportion of cases with proliferative endometrium on biopsy. In both fertile and infertile women, more than 7% of endometrial biopsies performed 7-13 days after a positive home urine LH test revealed proliferative endometrium. Patient interpretation of home urine LH test kits not uncommonly results in false-positive tests. Women planning menstrual cycle testing or procedures related to ovulation may benefit from additional confirmatory testing.

The unreliability of home urine luteinizing hormone test kits has important implications for clinical practice

A positive home urine LH kit test does not reliably indicate ovulation. This has important implications for clinical practice, whether these tests are used to confirm ovulation during a diagnostic evaluation, or used to time procedures such as artificial insemination.

Functional genomics of endometrial receptivity using an interceptive approach

Functional genomics of endometrial receptivity using an interceptive approach

Impact of controlled ovarian hyperstimulation on endometrial gene expression profiles during the window of implantation

Gene expression profiling of human endometrial receptivity has recently started and demonstrated that endometrial receptivity is an active process involving hundreds of up-/down-regulated genes. In the present work, we have investigated the impact of controlled ovarian hyperstimulation (COH) on endometrial gene expression in human endometrial receptivity. Endometrial samples were collected from ovum donors at days LH+2 (n=5) and LH+7 (n=5) as determined by urinary LH surge during a single natural cycle. Also, endometrial samples were obtained from the same patient at day LH+7 (n=5) of the natural cycle and at day hCG+7 (n=5) of the next cycle during COH used for IVF treatment. Genomic analysis of endometrial samples was performed using microarray technology. The protocol for ovarian stimulation consisted of a long protocol with leuprolide acetate with FSH/HP, hMG and hCG. Leuprolide acetate and gonadotropin injections were discontinued the day of hCG administration, and progesterone supplementation was not administered. Samples were hybridized onto the GeneChip HG_U133A (Affymetrix) encompassing more than 22,000 human DNA fragments, and the results were validated by RT-PCR and quantitative RT-PCR . Our study demonstrates that the gene expression in the COH hCG+7 samples is deregulated as compared to the natural cycle at LH+7. No luteal support was administered during COH for ovum donation, in order to get a clear comparison between natural and COH cycles. Genes regulated during the window of implantation in the natural cycle are more comparable with the LH+2 than with the LH+7 patterns in the COH cycle. This suggests a delay in the gene expression regulation that is required for the formation of a receptive endometrium in COH cycles. This remarkable finding justifies furthers research to identify COH protocols with minimal impact on endometrial development. Our study demonstrates that the gene expression in the COH hCG+7 samples is deregulated as compared to the natural cycle at LH+7. No luteal support was administered during COH for ovum donation, in order to get a clear comparison between natural and COH cycles. Genes regulated during the window of implantation in the natural cycle are more comparable with the LH+2 than with the LH+7 patterns in the COH cycle. This suggests a delay in the gene expression regulation that is required for the formation of a receptive endometrium in COH cycles. This remarkable finding justifies furthers research to identify COH protocols with minimal impact on endometrial development.

Reproductive physiology and development of artificial insemination technology in killer whales (Orcinus orca).

Research was conducted to define the basic reproductive physiology of killer whales (Orcinus orca) and to use this knowledge to facilitate the development of artificial insemination procedures. The specific objectives were 1) to determine the excretory dynamics of urinary LH and ovarian steroid metabolites during the estrous cycle; 2) to evaluate the effect of an exogenously administered, synthetic progesterone analog on reproductive hormone excretion; 3) to validate the use of transabdominal ultrasound for ovarian evaluation and timing of ovulation; 4) to examine the quality of semen after liquid storage and cryopreservation; and 5) to develop an intrauterine insemination technique. Based on urinary endocrine monitoring of 41 follicular phases and 26 complete cycles from five females, estrous cycles were 41 days long and comprised a 17-day follicular phase and a 21-day luteal phase. A consistent temporal relationship was observed between peak estrogen conjugates and the LH surge, the latter of which occurred approximately 0.5 days later. Two animals placed on oral altrenogest (three separate occasions for 30, 17, and 31 days, respectively) excreted peak urinary estrogen concentrations 25 days after withdrawal that were followed by sustained elevations in urinary pregnanediol-3alpha-glucuronide excretion. Mean preovulatory follicle diameter was 3.9 cm (n = 6), and ovulation occurred 38 h (n = 5) after the peak of the LH surge. Based on visual estimates of motility, liquid-stored semen maintained 92% of its raw ejaculate sperm motility index (total progressive motility x kinetic rating [0-5 scale, where 0 = no movement and 5 = rapid progressive movement]) when held at 4 degrees C for 3 days postcollection. Semen cryopreserved using a medium freezing rate demonstrated good postthaw total motility (50%), progressive motility (94%), and kinetic rating (3.5). Insemination during eight estrous cycles resulted in three pregnancies (38%), two from liquid-stored and one from cryopreserved semen. Two calves were delivered after gestation lengths of 552 and 554 days, respectively. These data demonstrate the potential of noninvasive endocrine monitoring combined with serial ultrasonography to improve our understanding of the reproductive biology of cetaceans. This fundamental knowledge was essential for ensuring the first successful conceptions, resulting in live offspring, using artificial insemination in any cetacean species.

Increasing the dose of human menopausal gonadotrophins on day of GnRH antagonist administration: randomized controlled trial

A significantly lower pregnancy rate following the gonadotrophin-releasing hormone (GnRH) antagonist protocol as compared with the long GnRH agonist protocol has been reported. The objective of this study was to investigate whether increasing the dose of gonadotrophins on the day of antagonist administration would increase the pregnancy rate. This study is an open labelled, randomized controlled trial and allocation was done using sealed envelopes. One hundred and fifty-one subfertile couples undergoing IVF/intracytoplasmic sperm injection (ICSI) cycles were included in the study. Ovarian stimulation was started on day 3 of the cycle, using 150–300 IU human menopausal gonadotrophin (HMG)/day. From day 8 onward, daily vaginal ultrasound and daily urinary LH estimation were performed. If a premature LH rise was detected, the cycle was cancelled. The antagonist (0.25 mg daily) was started when the leading follicle reached 15 mm in mean diameter and LH testing in urine was negative up to and including the day of human chorionic gonadotrophin (HCG) injection. Patients were randomized on the day of starting the antagonist into two groups: group A, 72 patients with no increase in HMG dose, and group B, 79 patients in whom the dose of HMG was increased by 75 IU on the day of antagonist administration, and continued till the day of HCG administration. The results showed no statistically significant difference between the groups regarding number of oocytes retrieved, embryos obtained, implantation rate, clinical pregnancy rate and multiple pregnancy rate. It was concluded that there is no clinical evidence for increasing the dose of HMG on the day of antagonist administration.