TUNEL labeling with BrdUTP/anti-BrdUTP greatly underestimates the level of sperm DNA fragmentation in semen evaluation.

Many studies have now confirmed that sperm DNA fragmentation (SDF) is associated with a poorer outcome of some forms of assisted reproduction technology. For this reason, SDF is an important parameter to evaluate in male fertility assessment. TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay coupled to flow cytometry is one of the most promising methods for SDF quantification.

Endometrial thickness influences neonatal birth weight in pregnancies with obstetric complications achieved after fresh IVF-ICSI cycles.

Purpose: Pregnancy-associated complications, duration of gestation and parity are well-known predictors of neonatal birth weight. Assisted reproductive technology (ART) affects neonatal birth weight as well. Endometrial thickness as measured on the day of HCG triggering may therefore impact on the neonatal birth weight.

Isolation of spermatozoa with low levels of fragmented DNA with the use of flow cytometry and sorting.

Objective: To develop a method to select spermatozoa with low DNA fragmentation rates for assisted reproduction technologies (ART).

Design: Multistep prospective cohort study.

Setting: University hospital.

First successful pregnancy in Switzerland after prospective sex determination of the embryo through the separation of X-chromosome bearing spermatozoa.

Question Under Study: The feasibility and the potential advantages of separating X-chromosome bearing spermatozoa for the prevention of a severe X-chromosome linked disorder with the use of intracytoplasmic sperm injection are presented.

Method: A carrier of muscular dystrophy type Becker was treated with intracytoplasmic sperm injection, using spermatozoa previously stained with the Hoechst dye 33342 and sorted with flow cytometry.

Results: After transfer of one single blastocyst, an intrauterine pregnancy arose.

[Present-day reproductive medicine and future possibilities].

Despite the controversial discussions, which have accompanied it since its first introduction of into clinical medicine particularly in Switzerland, assisted reproductive medicine has come to acquire a firm standing in present-day society. This broad acceptance is the logical consequence of the availability of efficient methods of family planning which started with the introduction of oral contraception in the early sixties. Over time assisted reproductive medicine has grown with a considerable degree of efficacy due to improvements in the culture conditions in the embryology laboratory and due to sophisticated ovarian hyperstimulation.

[Preservation of fertility in tumour patients].

The success rates of present-day chemotherapy have provoked a rising awareness with regard to the preservation of quality of life among successfully treated patients. Among other factors, quality of life also implies the capacity to procreate. Unfortunately, both in men and women chemotherapy often irreversibly destroys the production of gametes, thereby causing permanent infertility.

The multipotency of luteinizing granulosa cells collected from mature ovarian follicles.

Graafian ovarian follicles consist of follicular fluid, one single mature oocyte, and several hundred thousands of granulosa cells (GCs). Until now, luteinizing GCs have been considered to be terminally differentiated, destined to undergo death after ovulation. Present concepts of luteal function, endocrine regulation of early pregnancy, and the recruitment of new ovarian follicles are all based on the cyclical renewal of the entire population of GCs.

First delivery of healthy offspring after freezing and thawing of oocytes in Switzerland.

The interest in long-term storage of uninseminated oocytes through cryopreservation has seen a recent upsurge, because it provides the potential to assist young women to postpone childbirth after having overcome a malignant disease or delaying childbirth until after management of a professional career. The low fertilisation rate of frozen/thawed oocytes in earlier feasibility trials can now be improved by using intracytoplasmic sperm injection (ICSI) for assisting the penetration of the spermatozoon through the oocyte's hardened zona pellucida. Another reason for the reported low success rates of oocyte cryopreservation in earlier studies may have been the low developmental potential of spare oocytes, which were available for experimental cryopreservation.

Elaboration of a working model for the involvement of inhibin A as a mediator of the preovulatory rise of progesterone levels.

During the final days of follicular development, exogenously administered follicle-stimulating hormone (FSH) produces a rise in serum progesterone level. The aim of the present study was to investigate the possible source and regulation of this preovulatory progesterone surge. Four sets of matching treatments with gonadotropins for in vitro fertilization and intracytoplasmic sperm injection were selected from a cohort of 953 treatments in 244 couples.

Comparative birth weights of singletons born after assisted reproduction and natural conception in previously infertile women.

Background: The possible interference of assisted reproduction techniques (ART) with epigenetic reprogramming during early embryo development has recently sparked renewed interest about the reported lower birth weight among infants born as a consequence of infertility treatments. However, the latter finding so far has relied on the comparison of the birth weight of infants conceived with ART to general population data. A more appropriate comparison group should involve pregnancies in infertile women after natural conception.

Apoptosis and differentiation induced by staurosporine in granulosa tumor cells is coupled with activation of JNK and suppression of p38 MAPK.

We report here that staurosporine can induce apoptosis or differentiation of granulosa tumor cells depending on its dosage. In presence of staurosporine concentrations > 50 nM, apoptosis was triggered in human granulosa cell tumor cells COV434. In the presence of concentrations < 50 nM, the shape of the otherwise globular granulosa cells differentiated into a flattened epithelioid-like appearance.

Progesterone serum levels during the follicular phase of the menstrual cycle originate from the crosstalk between the ovaries and the adrenal cortex.

Background: The preovulatory rise of progesterone is important for ovulation, but both its regulation and its origin are controversial. Three experiments were performed to determine whether follicular phase progesterone arises from the ovary, the adrenal cortex or both.

Methods: The first study was performed in patients scheduled for assisted reproduction, who received a long-acting GnRH agonist either during intake of an oral contraceptive or during the luteal phase of an otherwise untreated menstrual cycle.

Characterization of an immortalized human granulosa cell line (COV434).

We have investigated the biological characteristics of an immortalized granulosa cell line (COV434), which may be used to study follicular and oocyte maturation in vitro. Granulosa cell function was defined as consisting of three distinct properties: (i) production of 17beta-oestradiol in response to follicle stimulating hormone (FSH); (ii) presence of specific molecular markers of apoptosis enabling the induction of follicular atresia; and (iii) capacity to form intercellular connections with cells surrounding an oocyte. The addition of FSH to the culture medium supplemented with 10% fetal calf serum and 4-androstene-3,17-dione resulted in proliferation of the COV434 granulosa cells and in an increased synthesis of 17beta-oestradiol, indicating the presence of the FSH receptor and cytochrome P450 aromatase in these cells.

Prospective evaluation of the ultrasound appearance of the endometrium in a cohort of 1,186 infertile women.

Objective: To investigate the effect of differences in endometrial thickness and pattern as visualized with present-day high-resolution transvaginal ultrasound equipment on the outcome of assisted reproductive treatment.

Design: Prospective cohort study.

Setting: Two large infertility units in university hospitals.

[Desire for children in tumor patients].

After a chemo- and/or radiotherapy not only acute side effects but also longterm side effects do occur. The following longterm side effects are observed: irregularities in the menstrual cycle, early onset of menopause and infertility. They are of special importance to children, teenagers and young adults having survived a malignancy.

Diagnostic accuracy of computer-assisted sperm motion analysis.

One retrospective and two prospective studies were conducted among 218 couples treated with in-vitro fertilization (IVF) to establish the reproducibility and diagnostic accuracy of computer-assisted sperm analysis (CASA) with swim-up spermatozoa for the prediction of the fertilization rate of oocytes in vitro. Based on the results of a preliminary retrospective analysis in 49 patients, the 'curvilinear velocity' (VCL) was chosen as the most distinctive motion parameter of sperm function and the median was used to represent the entire sperm population. The number of inseminated motile spermatozoa was then adjusted to median VCL during two subsequent prospective studies with clinical IVF.

Studies on the origin of redox enzymes in seminal plasma and their relationship with results of in-vitro fertilization.

Glutathione (GSH), GSH peroxidase (GPX), GSH reductase (GRD), superoxide dismutase (SOD) and catalase-like enzyme activity were quantified in seminal plasma from normozoospermic patients, men with known distal ductal occlusion, proven fathers and male partners of couples receiving in-vitro fertilization (IVF) treatment for both male and female causes. Glutathione was non-detectable (< 2.5 microM) in seminal plasma.

Low multiple pregnancy rates and reduced frequency of cancellation after ovulation induction with gonadotropins, if eventual supernumerary follicles are aspirated to prevent polyovulation.

Purpose: Our purpose was to demonstrate the feasibility of the routine aspiration of supernumerary follicles in infertile patients with imminent polyovulation after ovulation induction with gonadotropins and to examine its effect on the frequency of cycle cancellation and on the (multiple) pregnancy rate.

Methods: The data on 796 treatment cycles, performed between 1989 and 1996 on 410 infertile couples, were analyzed retrospectively. From October 1992, whenever necessary, supernumerary ovarian follicles were selectively aspirated transvaginally under ultrasound guidance to prevent the ovulation of more than three follicles.

Production of reactive oxygen species by and hydrogen peroxide scavenging activity of spermatozoa in an IVF program.

Purpose: Reactive oxygen species (ROS) including H2O2 produced by spermatozoa have been suggested, on one hand, to be associated with idiopathic male infertility and, on the other hand, to stimulate certain sperm function leading to fertilization. The influence of ROS on fertilization was investigated in 75 IVF patients by correlating fertilization rates with the production of ROS and the H2O2-scavenging activity of swim-up spermatozoa prepared in parallel with the IVF samples.

Results: Low rates of ROS production by the swim-up sperm was detected by the luminol-dependent chemiluminescence assay.

Experience with transvaginal ultrasound-guided aspiration of supernumerary follicles for the prevention of multiple pregnancies after ovulation induction and intrauterine insemination.

Objective: To avoid multiple pregnancies caused by ovulation induction.

Setting: Infertile couples treated in the Women's Hospital and the Institute of Reproductive Medicine of the University of Münster, Münster, Germany.

Design: The outcome of ovulation induction in patients in whom supernumerary ovarian follicles were aspirated transvaginally was compared with the outcome in patients in whom this intervention was not necessary.

Spontaneous recrudescence of spermatogenesis in the photoinhibited male Djungarian hamster, Phodopus sungorus.

Photosensitive rodents exposed to inhibitory short photoperiods become insensitive to this environmental factor after prolonged exposure. During the following process of spontaneous recrudescence, the animals that have adapted to the winter season show a return of all seasonal parameters. In the Djungarian hamster, obvious photoperiod-dependent changes are reinitiation of the reproductive organs, a 20-30% increase in body weight, and a moult from whitish fur into brown summer fur.

[Recombinant growth hormone for support of ovarian gonadotropin treatment in a hypophysectomized patient: a case-control study].

In a patient, suffering of apituitarism after the surgical removal of a cranyopharyngeoma at the age of 14, was treated with various doses of human urinary gonadotropin preparations for in vitro fertilization and embryo transfer. The first five treatments were performed without the additional administration of recombinant growth hormone, but due to the administration of increasing doses of exogenous gonadotropins a pregnancy was finally achieved leading to the birth of a healthy girl. Later, as the couple desired a second baby, the treatment was repeated using a low dose of gonadotropins in combination with recombinant growth hormone.

Pulsatile subcutaneous versus bolus intramuscular gonadotrophin administration after pituitary suppression with a long-acting gonadotrophin-releasing hormone analogue: a controlled prospective study.

The potential advantages of pulsatile s.c. administration instead of daily bolus i.

Peroxidase-positive round cells and microorganisms in human semen together with antibiotic treatment adversely influence the outcome of in-vitro fertilization and embryo transfer.

Human semen contains not only spermatozoa but also other cells routinely differentiated as being peroxidase-positive (e.g. leucocytes) and peroxidase-negative (e.