Vera Baukloh

Pollutants in human follicular fluid

The present study was initiated to clarify whether environmental pollutants can reach the human follicular fluid, an important biologic component for reproduction, in concentrations which could be harmful to the gamete. Fifteen patients with tubal malformations were treated in an in vitro fertilization program whereby 18 follicle aspirates were obtained for investigation. The substances which were measured in this study were alpha-, beta-, and gamma-hexachlorocyclohexane (HCH), dichlorodiphenyltrichlorethane (DDT), polychlorinated biphenyls (PCB), hexachlorobenzene (HCB), dieldrin, and heptachlorepoxide (HepE). None of the patients became pregnant during the treatment cycle.

Human ovum recovery via operative laparoscopy and in vitro fertilization

Between January 1979 and December 1980, 142 laparoscopies were performed on 114 patients with long-standing tubal factor infertility. Twenty-eight were performed during spontaneous menstrual cycles, 20 following continuous human menopausal gonadotropin/human chorionic gonadotropin (hMG/hCG) therapy, and 94 following intermittent hMG/hCG stimulation. Follicular ripeness was judged by multiple criteria, which were also used to time the laparoscopy. Operative procedures were performed in all cases and follicular puncture was attempted in all but 16 subjects, where adhesions prevented access to the ovaries. In 28 spontaneous cycles, 28 follicles were punctured and 17 ova recovered; whereas in the 98 patients where ovulation was stimulated, 217 follicles were punctured and 43 ova collected. Following in vitro fertilization with the husbands spermatozoa and embryo culture, pronucleus formation, 2-cell, 4-cell, 8-cell, and 16-cell stages were observed in 18 oocytes obtained from 17 patients. Embryo transfer has, however, not yet been performed.

Monoclonal Sperm Antibodies: Their Potential for Investigation of Sperms as Target of Immunological Contraception

ABSTRACT: We generated 149 hybridoma cell lines secreting antibodies against human spermatozoal antigens of which antibodies from 136 hybridoma lines also reacted with seminal plasma constituents. The occurrence of common antigeneic determinants on spermatozoa and seminal plasma was further demonstrated by competitive inhibition ELISA tests. We found that seven hybridoma clones secreted antibodies reactive with spermatozoa but nonreactive with seminal plasma. Antibodies from 5 clones were sperm‐agglutinating and from 15 clones sperm‐immobilizing. Localization of sperm antigens reactive with the monoclonal antibodies was demonstrated by indirect immunoperoxidase staining. A synthetic decapeptide, earlier shown to be reactive with naturally occurring human iso‐ and autosperm antibodies, was shown to be reactive with the monoclonal antibody VII‐5 in ELISA tests.

Pregnancy after vitrification of pronuclear stage oocytes biopsied for polar body aneuploidy screening

This paper reports the case of a patient undergoing IVF and polar body analysis for aneuploidy screening because of advanced maternal age. A total of nine fertilized oocytes were obtained from the patients third treatment cycle. Three supernumerary pronuclear stage oocytes were identified as suitable after aneuploidy screening for five chromosomes and vitrified. Because the fresh cycle did not result in a pregnancy, vitrified pronuclear stage oocytes were warmed for transfer in an artificial cycle. All three zygotes survived warming and were transferred to the patients uterus 2 days later as 4-, 5- and 6-cell stage embryos. After 2 weeks, a positive pregnancy test indicated successful implantation. The gestation is developing normally and is presently (end of September 2007) in the 22nd week.

Intra-age, intercenter, and intercycle differences in chromosome abnormalities in oocytes.

OBJECTIVE To determine the extent of intra-age and intercycle variations in the frequency of first polar body aneuploidy in two consecutive cycles of oocyte retrieval undertaken by the same patient within 1 year. DESIGN Retrospective study. SETTING Fertility centers. PATIENT(S) Infertile couples undergoing IVF. INTERVENTION(S) Patients underwent two consecutive cycles of preimplantation genetic screening through first polar body biopsy within 1 year. MAIN OUTCOME MEASURE(S) Meiosis I aneuploidy. RESULT(S) A total of 226 patients underwent 452 cycles of preimplantation genetic screening. Differences within age groups were wide, with 0-100% of oocytes being chromosomally normal in all age groups. Euploidy rates between centers were significantly different (48% vs. 25%). Intercycle differences for the same patient were also wide (0-100%), but with 68.5% of patients having less than ±2 euploid eggs of difference between cycles. CONCLUSION(S) Although euploidy rate decreased on average with advancing maternal age, the high intra-age and intercenter variation in oocyte chromosome abnormalities emphasize the difficulty in estimating how many euploid oocytes a specific woman will have. This may have repercussions for fertility preservation where a defined number of eggs are currently frozen just based on maternal age.

Induction of High Titer Mouse-Antihuman Spermatozoal Antibodies by Liposome Incorporation of Spermatozoal Membrane Antigens

ABSTRACT: This is the first report of induction of high titer mouse‐antihuman spermatozoal antibodies using a spermatozoal antigen incorporated in liposomes. The peptide antigens that were incorporated into pure phosphatidylcholine liposomes are known to react with the naturally occurring antibodies in sera of sterile patients. Electron microscopy and the complement‐dependent glucose release tests indicated that the peptides were entrapped in the aqueous phase between the lipid bilayers in the liposome vehicles. This is in contrast to most other liposomal antigens that are expressed on the outer lipid surface. In view of the nontoxic nature of the liposomes these results could be of some interest in immunological fertility regulation trials with appropriate antigens in an allogeneic system.

Patterns of serum-luteinizing hormone surges in stimulated cycles in relation to injections of human chorionic gonadotropin**Supported by Serono Diagnostics, Freiburg, FRG.

Endogenous-luteinizing hormone (LH) surges may complicate the management of in vitro fertilization cycles. To investigate the effects of LH surges after hormonal stimulation 53 IVF cycles were analyzed by assessing LH levels three times daily until egg collection. In 43% the LH rise started before the planned exogenous trigger for ovulation was given, in 11% the rise occurred simultaneously with and in 45% after the injection of human chorionic gonadotropin. Three main patterns of serum LH surges were identified: (A) low-LH tonus with straight increase to maximum; (B) low tonus with elevation before straight increase; (C) high tonus with large variations but no prominant peak. These patterns were not related to the follicular estradiol increase, luteal steroid concentrations or resulting pregnancy rates.

Collection of Human Eggs for In Vitro Fertilization

The conjugation of male sperm and female eggs, and the entire intrauterine development of mammals and mankind under normal physiological conditions take place in complete darkness. Man sees light for the first time at birth. With the technique of in vitro fertilization we deviate from these physiological conditions. The gametes are exposed to light, at the time of ovum pick up when the bright light of the endoscopist is shed on the follicle, at the time of masturbation while the semen is collected in a plastic container, and also when the egg is inseminated in culture medium under the microscope (even when using as little light as possible). It therefore seems impossible to create optimal conditions for extracorporeal fertilization. Human ovulation is nearly independent of external conditions and represents, compared to ovulation dependent on external circumstances as in the rabbit or cat, a certain effect of domestication. In other mammals, delayed fertilization and implantation allow the delivery of new life under optimal life conditions. As examples we can use the embryonic dormancy of the deer, kangaroo, and koala bear. The human egg, however, has to be aspirated at a time which cannot be influenced to a great extent, and must be transferred at the right time in order to have a slight chance of implantation.