H. van der Ven

Reproductive dysfunction in women with epilepsy: recommendations for evaluation and management

Background: Epilepsy is commonly associated with reproductive endocrine disorders. These include polycystic ovary syndrome (PCOS), isolated components of this syndrome such as polycystic ovaries, hyperandrogenaemia, hypothalamic amenorrhoea, and functional hyperprolactinaemia. Objective: To summarise the currently known relations between epilepsy and reproductive endocrine disorders. Methods: A review of clinical experience and published reports. Results: The most likely explanations for endocrine disorders related to epilepsy or antiepileptic drugs are: (1) a direct influence of the epileptogenic lesion, epilepsy, or antiepileptic drugs on the endocrine control centres in the brain; (2) the effects of antiepileptic drugs on peripheral endocrine glands; (3) the effects of antiepileptic drugs on the metabolism of hormones and binding proteins; and (4) secondary endocrine complications of antiepileptic drug related weight changes or changes of insulin sensitivity. Regular monitoring of reproductive function at visits is recommended, including questioning about menstrual disorders, fertility, weight, hirsutism, and galactorrhoea. Particular attention should be paid to patients on valproate and obese patients or those experiencing significant weight gain. Single abnormal laboratory or imaging findings without symptoms may not constitute a clinically relevant endocrine disorder. However, patients with these kinds of abnormalities should be monitored to detect the possible development of a symptomatic disorder associated with, for example, menstrual disorders or fertility problems. Conclusions: If a reproductive endocrine disorder is found, antiepileptic drug treatment should be reviewed to ensure that it is correct for the particular seizure type and that it is not contributing to the endocrine problem. The possible benefits of a change in treatment must be balanced against seizure control and the cumulative side effect of alternative agents.

Oocyte zona birefringence intensity is associated with embryonic implantation potential in ICSI cycles

A retrospective study recently showed that oocytes presenting with a high birefringence of the inner zona layer were more often associated with conception cycles. To further investigate these findings, a prospective study was conducted between September 2005 and September 2006 including intracytoplasmic sperm injection (ICSI) cycles presenting with at least two embryos for transfer. Using a polarization imaging system, oocytes were classified prior to ICSI treatment as having either a high zona birefringence (HZB) or a low zona birefringence (LZB) of the zona pellucida. Using zona birefringence as the only selection criterion, two fertilized oocytes, preferably derived from HZB oocytes, were selected for further culture and transfer. The required criteria were met by 135 ICSI cycles (124 patients; 34.9 +/- 4.1 years of age). Embryos for transfer were used in 20 cycles derived from HZB/HZB oocytes, in 50 cycles from HZB/LZB oocytes and in 65 from LZB/LZB oocytes. The corresponding implantation (P < 0.025), pregnancy (P < 0.005) and live birth (P < 0.025) rates were significantly different between HZB/HZB and HZB/LZB versus LZB/LZB group. Embryo development was superior in embryos derived from HZB oocytes. This study concludes that oocyte zona birefringence is a good selection criterion and a good predictive criterion for embryo implantation potential.

Oocyte assessment and embryo viability prediction: birefringence imaging

Embryo viability is a key element for success in assisted reproduction. Since the beginning of the era of assisted reproduction treatment, embryo viability was mostly considered to be a function of developmental progression during the preimplantation phase. In the last decade, several morphological criteria of oocytes and embryos were evaluated with regard to their potential for predicting embryo viability. The introduction of polarization light microscopy systems in assisted reproduction has enabled the detection of structures within oocytes that possess a natural birefringence. Birefringence imaging of the meiotic spindle and the zona pellucida in living animal and human oocytes represents a new approach in the assessment of oocyte and embryo viability. The technique was applied in several studies to select oocytes in order to improve treatment success. This review will summarize the present knowledge of birefringence imaging. The various applications in basic and clinical research as well as in clinical treatment will be presented, especially with regard to their effect on assisted reproduction.

Polar body biopsy: a viable alternative to preimplantation genetic diagnosis and screening

Polar body diagnosis (PBD) is a diagnostic method for the indirect genetic analysis of oocytes. Polar bodies are by-products of the meiotic cell cycle, which have no influence on further embryo development. The biopsy of polar bodies can be accomplished either by zona drilling or laser drilling within a very short time period. However, the paternal contribution to the genetic constitution of the developing embryo cannot be diagnosed by PBD. The major application of PBD is the detection of maternally derived chromosomal aneuploidies and translocations in oocytes. For these indications, PBD may offer a viable alternative to blastomere biopsy as the embryos integrity remains unaffected, in contrast to preimplantation genetic diagnosis (PGD) by blastomere biopsy. The rapid pace of developments in the field of molecular diagnostics will also influence the advantages of PBD, and probably allow more general diagnostic applications in the future.

The role of ovarian volume in an in vitro fertilization programme as assessed by 3D ultrasound.

Abstract The study was designed to investigate the role of ovarian volume, as assessed by three-dimensional (3D) sonography, in predicting conception in an in-vitro fertilization-embryo transfer (IVF-ET) programme. Transvaginal 3D sonography was performed in 152 cycles before initiation of ovarian stimulation (day 1) and on the day of oocyte retrieval. Ovarian volume showed no significant correlation with IVF outcome. On the contrary, all ovarian measurements were lower, albeit non-significantly, in the conception group. Fifteen patients (15/152, 9.9%) had a minimum unilateral ovarian volume of ≤3 mL (1 SD below the mean) on day 1 of the stimulation cycle. In this subgroup, the likelihood of conception was 6.7% (1/15) versus 21.9% (30/137) in patients with an initial minimum ovarian volume of >3 mL. This difference did not reach statistical significance. In both groups, cancellation rates due to poor ovarian response or lack of fertilization were similar. In conclusion, ovarian volumetry as assessed by three-dimensional ultrasound failed to predict conception in women undergoing IVF treatment.

LASER- ASSISTED CRYOPRESERVATION OF SINGLE HUMAN SPERMATOZOA IN CELL- FREE ZONA PELLUCIDA

Summary. Improved procedure for efficient cryopreservation of single human spermatozoa in cell‐free human zona pellucida is reported. We used a diode laser system for efficient and precise creation of a single hole into the zona pellucida of a degenerated or immature human oocyte. This allowed the extraction of the cytoplasm using a micropipette with a diameter of 10–15 μm. Through the same opening, human spermatozoa were inserted into the empty zona. We used motile and laser immobilized spermatozoa. Immobilized sperm were obtained by a single laser irradiation delivered in the vicinity of the sperm tail prior to insertion. This new immobilization procedure was shown to have no deleterious effect on membrane integrity and sperm viability. Following sperm transfer into the zona, the laser‐drilled hole was closed with an oil droplet which was expelled from the micropipette during withdrawal to avoid loss of spermatozoa. This facilitated detection of the otherwise translucent zona during the cryopreservation procedure. After thawing, all cryopreserved zonae (20/20) could be successfully retrieved. Spermatozoa were recovered from the zona pellucida through the hole used for insertion. The rate of sperm recovery for initially motile spermatozoa was 80% vs. 92% for laser immobilized spermatozoa. Sperm viability was 81% and 84%, respectively, detected by a Hoechst stain. This technique makes cryopreservation of single human spermatozoa easy and feasible and appears beneficial for couples with severe male infertility and for those facing repeated surgical sperm extraction.

Recovery of ejaculated spermatozoa for intracytoplasmic sperm injection after anti-inflammatory treatment of an azoospermic patient with genital tract infection : a case report

In this paper our experiences with anti‐inflammatory treatment of an infertile patient with azoospermia and concomitant silent genital infection are reported. The patient was referred to our fertility centre with prediagnosed non‐obstructive azoospermia and no spermatozoa were found in the ejaculate on two occasions. The patient showed leukocytospermia and was suspected to be affected by genital infection. Therefore, anti‐inflammatory treatment was initiated and 8 weeks later examination of the ejaculate revealed a decreased number of leukocytes and the presence of few but motile spermatozoa. Subsequently, two ICSI cycles were performed with anti‐inflammatory therapy in parallel and a sufficient number of spermatozoa could be retrieved for injection. However, in a third cycle without previous treatment, examination of the ejaculate again revealed azoospermia and leukocytospermia.

Presence and localization of oscillin in human spermatozoa in relation to the integrity of the sperm membrane

We investigated the presence and localization of oscillin in human spermatozoa in relation to the integrity of the sperm membrane, which was assessed by the hypo‐osmotic swelling (HOS) test. We found no gross differences in the presence of oscillin in semen samples from men who presented with 70%, 40%, 25% or 2% of membrane‐intact spermatozoa. By immunofluorescence, membrane‐intact (HOS‐positive) spermatozoa showed staining of a single band at the equatorial region, whereas over 80% of HOS‐negative spermatozoa consistently showed a diffuse distribution of oscillin over the sperm head. However, some individuals presented with up to 50% of HOS‐positive spermatozoa showing an aberrant localization of oscillin. We found a significant correlation rate (r=0.70, P<0.05) between the percentage of HOS‐positive spermatozoa with an equatorial oscillin localization and the fertilization rates achieved after intracytoplasmic sperm injection. These data suggest that the localization of oscillin in human spermatozoa might have an impact on egg activation and fertilization rates.

Three-dimensional power Doppler ultrasound of the subendometrial blood flow under the administration of a contrast agent (Levovist).

MethodsBased on the results from 42 IVF patients on the day of oocyte aspiration, our study tested the hypothesis that 3-D Doppler ultrasound of the endometrium supported by a contrast agent possesses advantages over a conventional 3-D Doppler ultrasound examination. We also determined whether the subendometrial blood flow indices measured under continuous infusion of Levovist (Schering, Berlin, Germany) were an indicator of a successful implantation.ResultsIt was demonstrated that, after the administration of Levovist, all subendometrial blood flow parameters (e.g., vascularization index [VI: 8.82±16.64], flow index [FI: 35.67±9.33], and vascularization flow index [VFI: 3.99±8.80]) were significantly higher than without the contrast agent (p<0.001; VI: 3.09±7.11; FI: 29.13±14.26; VFI: 1.35±3.31). However, no significant differences between blood flow with contrast agent and pregnancy rate were found.ConclusionIn summary, quantitative 3-D Doppler ultrasound employing Levovist (Schering, Berlin, Germany) is significantly more sensitive when compared to conventional 3-D Doppler examination. For the first time this was demonstrated on 42 IVF patients on the day of oocyte aspiration. However, no correlation with outcome of the IVF-treatment could be found.

Einfluß von Pestiziden auf die Funktion von Spermatozoen in vitro

Die Diskussion uber einen moglichen Zusammenhang zwischen Umweltschadstoffen und einer Beeintrachtigung der Spermaqualitat stutzt sich bisher weitgehend auf statistische Daten (Schill 1986). In der vorliegenden Studie wurde nun der direkte Einflus von verschiedenen chlorierten Kohlenwasserstoffen auf wichtige Spermafunktionen in vitro untersucht. Normozoosperme Samenproben wurden in BWW Medium gewaschen (500 × g, 5 min) und anschliesend in BWW (Kontrolle 1) BWW + 1% DMSO (Kontrolle 2) und BWW + Toxin (DMSO max. 1%) resuspendiert (10 × 106 Spermien/ml). Die eingesetzten Toxine waren HCB (Hexachlorbenzol) DDE, Clophen (Gemisch verschiedener PCBs) und ein einzelnes Congener des PCB (22′66′ Tetrachlorobiphenyl). Die Spermasuspensionen wurden 2–12 Stunden inkubiert, wobei die Toxinkonzentration 0,1 – 10000 ng/ml betrug. Nach verschiedener Inkubationsdauer wurden folgende Spermaparameter beurteilt: Motilitat, Vitalitat, akrosomale Integritat (triple stain) und die funktionelle Membranintegritat (HOS-Test). Unter den beschriebenen in vitro Bedingungen zeigten PCB, Clophen, HCB und DDE einen meist zeit- und dosisabhangigen negativen Einflus auf die untersuchten Spermafunktionen. Die in dieser Studie verwendeten Substanzen sind unterschiedlich wirksam in bezug auf die Spermaqualitatseinschrankung (z.B. PCB insbesondere Motilitatsein- schrankung, DDE insbesondere Vitalitatseinschrankung und Induktion der akrosomalen Reaktion).