Ultrastructural analysis of TESE and semen sperm tails from patients exhibiting absolute immotility on the day of ICSI.

Sperm flagellar pathology was found to be the underlying cause of motility disorders that lead to male infertility. Conventional in vitro fertilization (IVF) procedures will fail when sperm show a total absence of motility. In such difficult cases intracytoplasmic sperm injection (ICSI) is the only available technique to fertilize an oocyte. Fertilization rates are low and may also be reduced when immotile sperm are used for ICSI from ejaculate of other than epididiymal or testicular origin. Presence of totally immotile sperm in the ejaculate on the day of ICSI if spermatogenesis is normal testicular sperm recovery can improve ICSI outcomes. But for patients having severe morphological or functional sperm defects embryos of lower quality tend to be produced when totally immotile sperm are used. In this study the 2 patients exhibiting totally immotile sperm in their ejaculates and TESE samples on the day of ICSI showed the same ultrastructural abnormalities. Peri-axonemal and axonemal abnormalities that were seen in association with sperm nucleus structural defects suggested that the source of sperm has no effect on morphologic characteristics and also reflects abnormality in both spermatogenesis and spermiogenesis. In this study the two patients who presented with oligoteratozoospermia with total immotility, using either ejaculate or TESE sperm fertilization and embryo development, can be obtained with ICSI, but no pregnancies were established after embryo transfers.

The impact of the source of spermatozoa used for ICSI on pronuclear morphology.

BACKGROUND: The aim of this prospective study was to find out whether the source of spermatozoa used for intracytoplasmic sperm injection (ICSI) has an impact on the morphological features of pronucleate zygotes, which make up the basis of a pronuclear scoring system for the selection of the most viable embryos for transfer. METHODS AND RESULTS: The study group consisted of 194 two pronucleate (2PN) ICSI zygotes, of which 144 originated from ejaculated (ES) and 50 from testicular spermatozoa (TS). At 18 h postinjection, 2PN zygotes were assessed for pronuclear alignment, polarity in nucleoli and cytoplasmic appearance; all of which were found to exhibit similar patterns of distribution between the ES and TS groups (P = not significant). At 25 h, the presence of first cleavage was similar for both groups; 11% of zygotes in the ES and 10% of those in the TS group underwent early cleavage (P = not significant). At 48 h, a quality score was obtained for cleaving embryos by multiplying the number of blastomeres with the grade of the embryo. Pronuclear scoring in both groups of spermatozoa correlated with embryo quality score at 48 h postinjection. There was a trend for a higher incidence of early cleavage and a lower incidence of pronuclear arrest with better pronuclear scoring embryos for both types of spermatozoa. CONCLUSION: The morphological features of pronucleate zygotes at 18 h after microinjection with ES and TS are similar to each other.

Gonadotropin-releasing hormone antagonist protocol: a novel method of ovarian stimulation in poor responders.

OBJECTIVE: To estimate the efficacy of gonadotropin-releasing hormone (GnRH) antagonist 'Cetrorelix' in poor responders comparing with the standard long protocol. DESIGN: The study population consisted of 21 poor responders who underwent ICSI and treated with Cetrorelix according to the multiple-dose protocol and who were compared with 21 poor responders treated according to the long protocol and who also underwent ICSI. Patients in both groups were matched for chronological age, the number of follicles found by ultrasound at the retrieval day and cause of infertility. Fifteen patients of GnRH antagonist group were treated with the combination of GnRH antagonist with clomiphene citrate (CC) plus gonadotropins, while six patients were treated with the combination of GnRH antagonist plus gonadotropins, but without CC. RESULTS: The use of GnRH antagonist in a multiple dose protocol gave a pregnancy rate of 14.28% which was in the range expected for patient with poor response, but with shorter treatment duration and with fewer ampoules of gonadotropins as compared with the use of a GnRH agonist protocol in a depot formulation. Within Cetrorelix group patients who received CC had a significant shorter duration of stimulation and needed fewer ampoules as compared with patients in the same group who did not receive CC. CONCLUSIONS: A GnRH antagonist multiple dose protocol may be the protocol of choice for the treatment of poor responders. The use of GnRH antagonist Cetrorelix ended with significantly less ampoules of gonadotropins and a shorter duration of stimulation.

Effect of the gonadotropin-releasing hormone antagonist ganirelix on cyclic adenosine monophosphate accumulation of human granulosa-lutein cells.

OBJECTIVE: To evaluate whether the GnRH antagonist ganirelix exerts an effect on cyclic adenosine monophosphate (cAMP) production of human granulosa-lutein (GL) cells in vitro. DESIGN: In vitro cell culture study. SETTING: Research laboratory of a university hospital. PATIENT(S): Mural GL and cumulus cells were obtained from 15 patients on whom controlled ovarian hyperstimulation was being performed for intracytoplasmic sperm injection treatment. INTERVENTION(S): Mural GL and cumulus cells were cultured for 48 hours with and without 1 nM ganirelix or triptorelin. For the last 6 hours, the cells were either exposed to 1-5 IU hCG or left unstimulated. MAIN OUTCOME MEASURE(S): At the end of the culturing period, the intracellular and extracellular cAMP accumulations were measured by an (125)I-scintillation proximity assay. RESULT(S): hCG induced dose-dependent increases in total cAMP accumulation. Stimulation with 1 IU/mL hCG resulted in 9-fold and 13-fold increases, and 5 IU/mL hCG resulted in 19-fold and 14-fold increases in total cAMP release from cumulus and mural GL cells, respectively. On the other hand, treatments with 1 nM GnRH antagonist ganirelix and 1 nM GnRH agonist triptorelin did not exert any significant changes on the basal and hCG-stimulated cAMP accumulation of mural GL cells and cumulus cells as compared with controls. CONCLUSION(S): Ganirelix does not influence basal and hCG-stimulated cAMP accumulation of human GL cells in vitro. cAMP is apparently not involved in the mechanism of action of GnRH analogs in human ovary.

Comparison of cryopreservation outcome with human pronuclear stage oocytes obtained by the GnRH antagonist, cetrorelix, and GnRH agonists.

This retrospective study was performed to examine the implantation and pregnancy rates of frozen-thawed pronuclear stage oocytes obtained with the use of a GnRH antagonist, Cetrorelix (Cetrotide((R)) ASTA-Medica, Frankfurt/M, Germany) used in a multidose protocol with hMG, and to compare these results with those obtained after a conventional long GnRH analogue protocol (Decapeptyl-Depot, Ferring, Kiel, Germany). The study population consisted of 31 infertile couples with frozen-thawed pronuclear stage oocytes after ICSI treatment using the GnRH antagonist Cetrorelix (Cetrorelix((R))) and 31 infertile couples with frozen-thawed pronuclear stage oocytes after ICSI treatment using the long GnRH analogue protocol. Patients underwent ICSI after down regulation with a GnRH agonist (Decapeptyl) and stimulation with hMG, or a GnRH antagonist (Cetrorelix) and hMG. The supernumerary pronuclear stage oocytes were cryopreserved and transferred in a later mildly stimulated cycle. The implantation and pregnancy rates for frozen-thawed pronuclear stage oocytes derived from the GnRH antagonist compared with the GnRH agonist were 3.26% versus 3.73% (P=1.0000) and 8.33% versus 10.25% (P=1.0000), respectively. To our knowledge we report here the first pregnancies obtained by the transfer of cryopreserved pronuclear stage embryos generated from ICSI using a GnRH antagonist in the collecting cycle. The use of Cetrorelix in a multiple dose protocol in combination with hMG does not demonstrate a negative effect on viability, implantation potential or pregnancy outcome as compared to 2PN conceptuses obtained from a long GnRH agonist-hMG protocol.

ICSI outcome in patients of 40 years age and over: a retrospective analysis.

OBJECTIVE: To report and analyse our experience with ICSI treatment in infertile women >/=40 years of age, with the intention of contributing to current debates on the effect of aging on the reproductive potential. STUDY DESIGN: 107 infertile couples in which the female partner was aged >/=40 years and who received ICSI treatment between January 1996 and December 1998. RESULTS: A total of 107 women underwent 171 treatment cycles during this period. Of 171 cycles initiated 33 were cancelled (cancellation rate=19.3%). In this way, 17 women did not have embryo transfer at all, while 90 patients had 138 cycles with oocyte retrieval and successful embryo transfer, with a mean number of embryos per transfer 2.36. Sixteen pregnancies occurred and eight of them ended in spontaneous abortion. The implantation rate was 4.9%, the pregnancy rate per initiated cycle was 9.35% and per transfer cycle 11.59%. The miscarriage rate was 50%. Moreover, 12 patients had supernumerary embryos, that were cryopreserved and transferred in 17 thawing cycles and resulted in two pregnancies ending in abortion. All pregnancies occurred when three embryos were available, except in two cases with two available embryos. The great majority of the total pregnancies (16 of 18) resulting in women aged between 40 and 42 years. CONCLUSION: Our data show that women 40 and older with existing ovarian function may benefit from ICSI treatment, even when the indication for treatment is male factor infertility. Supernumerary embryos, that are cryopreserved and transferred in subsequent cycles can improve the overall pregnancy rates per oocyte retrieval, although these women should be aware of the very high risk of miscarriage.

The outcome of intracytoplasmic sperm injection in patients with retrograde ejaculation.

The objective of this retrospective clinical study was to assess the benefit of assisted fertilization in cases of anejaculatory infertility due to retrograde ejaculation. We report the outcome of intracytoplasmic sperm injection (ICSI) treatment. In 16 couples in which the men suffered from retrograde ejaculation. We performed 35 cycles of ICSI with spermatozoa retrieved from post-ejaculatory urine. The patients had been instructed to alkalinize the urine by ingesting sodium bicarbonate before the procedure. The fertilization rate averaged 51.2%. Seven clinical pregnancies were achieved. Three spontaneous first trimester abortions occurred, but three live offspring were delivered and one pregnancy is ongoing. In conclusion, the use of ICSI may be feasible for patients with retrograde ejaculation who are resistant to medical treatment and whose sperm quality is so low or unpredictable that intrauterine insemination or conventional methods of in-vitro fertilization are not possible.

Pregnancies achieved after frozen-thawed pronuclear oocytes obtained by intracytoplasmic sperm injection with spermatozoa extracted from frozen-thawed testicular tissues from non-obstructive azoospermic men.

The use of frozen-thawed testicular tissue as a source of spermatozoa for intracytoplasmic sperm injection (ICSI) in non-obstructive azoospermia yields favourable fertilization and pregnancy rates while avoiding both repetitive biopsies and unexpected cycle cancellations. Spermatozoa were obtained from frozen-thawed testicular biopsy specimens from 67 non-obstructive azoospermic men. Following fertilization, supernumerary two pronuclear (2PN) oocytes were frozen. After thawing, 17 cycles of embryo transfer were carried out with a mean number of 2.7 embryos and a mean cumulative embryo score (CES) of 18.3 per transfer. The clinical pregnancy and implantation rates per transfer in these cycles (23.5 and 8.3% respectively) were comparable to those of fresh embryo transfers (35.7 and 12.7% respectively) with a mean number of 2.7 embryos and a mean CES of 28.7 per transfer. Abortion rates, although higher with cryopreserved 2PN oocytes were not significantly different. With this approach, cryopreservation of supernumerary 2PN oocytes can be used to improve the cumulative pregnancy rates in a severely defective spermatogenetic population. To our knowledge, these are the first pregnancies reported which have been obtained by the transfer of cryopreserved pronuclear oocytes obtained from ICSI using cryopreserved testicular spermatozoa.

Prenatal diagnosis of ostium secundum atrial septal defect by M-mode fetal echocardiography.

Advances in fetal echocardiography enable the prenatal diagnosis of a vast majority of congenital heart defects. On the other hand, ostium secundum atrial septal defect is difficult to diagnose in the fetus. Although a defect of minor clinical significance, this may be the only intrauterine finding of a chromosomal anomaly. We documented an abnormal oscillatory pattern of the foramen ovale flap motion by M-mode fetal echocardiography in 2 fetuses with ostium secundum atrial septal defect. This may be a more consistent finding for the prenatal diagnosis of ostium secundum atrial septal defect than the previously reported bidirectional flow across the foramen ovale on color Doppler sonography.

Doppler velocimetry of the uterine arteries after hysteroscopic rollerball endometrial ablation.

The aim of this study was to document the Doppler indices [pulsatility index (PI) and resistance index (RI)] of the uterine arteries in 30 patients who underwent hysteroscopic rollerball endometrial ablation for dysfunctional uterine bleeding by transvaginal pulsed Doppler sonography, and to reveal whether treatment failures (persistent menometrorrhagia) can be predicted by the blood flow characteristics of the uterine arteries in advance. On the basis of the outcome of patients at the end of the first postoperative year, the Doppler indices of the uterine arteries were meaningful 1 year after the operation when PI (1.32 +/- 0.11; mean +/- SD) and RI (0.71 +/- 0.04) in six menometrorrhagic patients were statistically different from PI (2.19 +/- 0.28; 1.95 +/- 0.36 and 1.82 +/- 0.37) and RI (0.87 +/- 0.06; 0.82 +/- 0.06 and 0.81 +/- 0.04) in normally menstruating, amenorrhoeic and hypomenorrhoeic patients respectively (P < 0.05). On the other hand, the patients who would be menometrorrhagic one year after the operation had a thicker endometrium in the first post-operative month. These findings suggest that the angiogenetic role of the persistent endometrial islands after endometrial ablation needs at some time to be reflected as changes in the Doppler parameters of the uterine arteries.

Osseous metaplasia of the endometrium: a rare cause of infertility and its hysteroscopic management.

This case report presents a very rare and long-standing cause of infertility, osseous metaplasia of the endometrium, and describes its successful management by hysteroscopy. A woman with a history of induced abortion 12 years ago, failed to conceive from that time on. The infertility work-up was unrevealing except for the presence of intracavitary calcification on ultrasonography. After diagnostic hysteroscopy, solid bony spicules covering the posterior wall of the endometrial cavity were removed by wire loop resectoscope. Histology established the diagnosis of osseous metaplasia of the endometrium. The patient conceived in her second spontaneous cycle and has an ongoing pregnancy at the time of writing. Hysteroscopy was an effective means of extracting this heterotopic tissue, thereby re-establishing fertility even after a long period of infertility.