The aim of this study was to identify gene expression patterns of the testis that correlate with the appearance of distinct stages of male germ cells. We avoided the pitfalls of mixed pathological phenotypes of the testis and circumvented the inapplicability of using the first spermatogenic wave as done previously on rodents. This was accomplished by using 28 samples showing defined and highly homogeneous pathologies selected from 578 testicular biopsies obtained from 289 men with azoospermia (two biopsies each). The molecular signature of the different developmental stages correlated with the morphological preclassification of the testicular biopsies, as shown by resampling-based hierarchical clustering using different measures of variability. By using analysis of variance (ANOVA) and extensive permutation analysis, we filtered 1181 genes that exhibit exceptional statistical significance in testicular expression and grouped subsets with transcriptional changes within the pre-meiotic (348 genes), post-meiotic (81 genes) and terminal differentiation (38 genes) phase. Several distinct molecular classes, metabolic pathways and transcription factor binding sites are involved, depending on the transcriptional profile of the gene clusters that were built using a novel clustering procedure based on not only similarity but also statistical significance.
In recent publications we have demonstrated that laparoscopic electrocautery of the ovarian surface (LEOS) is an effective method to reduce serum androgen concentrations, normalizing ovarian cycle length and the ovarian reaction to hormonal stimulation therapy in anovulatory patients with polycystic ovarian disease (PCOD). In this paper we show that these effects are not only temporary. Data from 206 patients undergoing LEOS and monitored for up to 72 months after surgery are evaluated; 145 patients achieved a total of 211 conceptions, giving a pregnancy rate of 70%, with a maximum of four conceptions in one patient. There were 38 early miscarriages (18%) and two late pregnancy losses as well as three ectopic implantations.
Prophylactic infusion of human serum albumin can reduce or mitigate severe ovarian hyperstimulation syndrome (OHSS) in patients at high risk. Recently, concern has been expressed in the lay press regarding the potential viral transmissions with blood constituents. Hence, we looked for a safe non-biological substitute with comparable physical properties in order to cope with this concern. One hundred patients of our in-vitro fertilization (IVF) programme with oestradiol serum concentrations> or = 11010 pmol/l on the day of human chorionic gonadotrophin injection and/or > or = 20 oocytes retrieved and/or previous severe OHSS were infused with 1000 ml 6% hydroxyaethyl starch solution at the time of oocyte collection and 500 ml 48 h later. A total of 82 IVF patients at risk without prophylactic infusions during the preceding years served as controls. Both groups were identical according to patient's age, body mass index, androgen concentrations, peak oestradiol concentrations, number of retrieved oocytes, fertilization and pregnancy rates. There were seven cases of severe OHSS in untreated patients and two cases in the treatment group (P = 0.08). In moderate OHSS a significant difference became obvious with only ten cases in the treatment group and 32 cases in the control group (P < 0.00001). Hydroxyaethyl starch solution seems to be an effective and economic alternative in reducing severe and moderate OHSS during IVF treatment.
Investigations into the effect of clomiphene citrate stimulation on endometrial differentiation were retrospectively performed in the mid-luteal phase on 21 patients undergoing ovulation induction prior to artificial insemination. A high incidence of irregular endometrial development was seen using morphological and morphometric criteria; in addition, increasing stromal oedema after clomiphene citrate treatment was observed. Measurable differences in glandular development (i.e. numbers of glands and glandular epithelial height) were found in early luteal phase biopsies (LH+2/+6) from patients treated with clomiphene citrate and normal fertile controls. Impaired endometrial development in the early luteal phase, which is a sensitive interval in preparation for implantation, might lead to an effective asynchrony between embryonic and uterine development and unfavourably influence the outcome of implantation.
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