B. Rosenbusch

DOPPLER SONOGRAPHIC FINDINGS AND THEIR CORRELATION WITH IMPLANTATION IN AN IN VITRO FERTILIZATION PROGRAM.

In 45 women from an in vitro fertilization (IVF) program, the uterine and ovarian blood flows were investigated by vaginal Doppler sonography. The resistance index was used to evaluate the blood pattern. When comparing the patients who became pregnant after embryo transfer (ET [group I, n = 12]) with those who did not conceive (group II, n = 33), it is evident that in group I the vascular resistance of the uterine arteries is significantly lower on the day of follicular aspiration. No differences could be detected in the ovarian vessels. The data obtained so far suggest that the receptivity of the endometrium is a crucial factor for successful implantation. In the final analysis, this can be appraised not only on the basis of morphological but also of hemodynamic parameters.

Influence of smoking on fertility in women attending an in vitro fertilization program

OBJECTIVE To investigate the influence of cigarette smoking of women on the fertilization and pregnancy rates obtained by IVF treatment. PATIENTS One hundred ninety-seven infertile, otherwise healthy women who entered an IVF program for the first time. SETTING Fertility unit at the Womens University hospital of the University of Ulm, Ulm, Germany. INTERVENTIONS The study population consisted of 197 women (23 to 39 years old) who were divided into the following groups: nonsmokers (n = 68), passive smokers (n = 26), and active smokers (n = 103) according to the cotinine concentration measured in follicular fluid. The reason for infertility was strictly a tubal factor with apparently normal ovulatory cycles. To guarantee an objective recording of tobacco smoke exposure, the smoking habit was not determined by questionnaires, but by cotinine, the principal metabolite of nicotine. RESULTS There were no significant differences in fertilization and pregnancy rates between the different groups. The E2 serum levels were decreased significantly in women who smoked when compared with the results obtained from nonsmokers and passive smokers. Overall, a strong negative correlation of the cotinine and E2 levels was observed (r = -0.65). CONCLUSION The results suggest that there is no clinically detectable impairment of fertilization potential due to female smoking and that there is a greater influence on the outcome of IVF by other factors.

Chromosomal analysis of unfertilized human oocytes prepared by a gradual fixation-air drying method

Two hundred and sixty-five unfertilized human metaphase II (MII) oocytes from an in vitro fertilization program were studied cytogenetically using our chromosomal technique, a gradual fixation-air drying method. Of the 265 oocytes, 185 (70%) were successfully karyotyped. There were 21 aneuploids (11.4%) consisting of 8 hyperhaploids (4.3%), 11 hypohaploids (5.9%) and 2 complex cases (1.1%). There were also 9 structural anomalies (4.9%) and 18 diploids (9.7%). In aneuploidy, the loss or gain of dyads (so-called nondisjunction) occurred more frequently than the loss or gain of monads (so-called predivision). The frequency of abnormally behaved chromosomes (segregation errors) due to nondisjunction, anaphase lag and predivision was studied among the seven chromosomal groups (A-G) and compared with the frequency expected from an equal probability of segregation errors in each of the 23 chromosomes. The observed frequency was somewhat higher than the expected frequency in groups E and G but the difference was not statistically significant in either group. These results were discussed in relation to previous studies on human M II oocyte chromosomes.

Cytogenetics of human spermatozoa: correlations with sperm morphology and age of fertile men

Sperm chromosomes from 15 fertile men were analyzed after fusion of their spermatozoa with zona-free hamster eggs. The total proportion of abnormal metaphases as well as the proportions of aneuploidy and structural aberrations were calculated for every man and examined for linear correlations with [1] sperm morphology and [2] the age of the persons studied. A positive correlation between the cytogenetic parameters and the percentage of abnormal sperm morphology was not evident, suggesting that assessment of sperm morphology cannot be used as an indicator of chromosomal damage in human spermatozoa. In contrast, there was a more distinct positive correlation between the age of donors and the three cytogenetic parameters studied.

Sperm chromosomes and habitual abortion

Genetic factors, especially numerical chromosome anomalies, play an important role in embryonic loss. Because somatic cell analysis cannot assess the risk of errors arising de novo during germ cell maturation, we investigated whether the male gametes from couples with habitual abortion carry a higher rate of anomalies than those from donors without reproductive dysfunction. Our results indicate that there is no significant difference between the two groups for the total rates of aneuploidy and structural anomalies. However, the levels of chromosome breaks and acentric fragments were significantly higher in the abortion group. The implications of this fact remain to be investigated.

Frequency and patterns of premature sperm chromosome condensation in oocytes failing to fertilize after intracytoplasmic sperm injection.

AbstractPurpose: The purpose of this study was to evaluate reasonsfor fertilization failure after intracytoplasmic sperminjection as a part of internal quality control and to reviewcorresponding previous data. Methods: One hundred injected but unfertilized oocytes werefixed and examined after Giemsa staining. Results: Three oocytes (3.0%) did not show the presenceof a spermatozoon and two (2.0%) contained pronuclearstructures. An intact spermatozoon was found in 25 cases(25.0%), whereas the sperm nucleus had undergonepremature chromosome condensation (PCC) in 70 cells (70.0%).A modified classification system was established tocharacterize the different PCC patterns. Conclusions: PCC indicates a correct intracytoplasmicinjection and excludes technical problems as a major reasonfor fertilization failure in the present study. A lack of oocyteactivation due to cytoplasmic immaturity is consideredresponsible for the occurrence of PCC. A review of theliterature shows that the role of sperm chromatinabnormalities in the process of nuclear decondensation needs furtherinvestigation.

Lack of correlation between ultrasonography and histologic staging of the endometrium in in vitro fertilization (IVF) patients.

In 53 patients of an in vitro fertilization (IVF) program with unsuccessful fertilization of oocytes, an endometrial biopsy was carried out on the day of the intended embryo transfer. The results were compared with the thickness (assessed on the very same day by means of ultrasonography) and the echo pattern of the endometrium, which was classified into four grades (A to D). We found grade A in 16 cases (30%), grade B in 22 cases (41.5%) and grade C in 15 cases (28.5%); no endometrium was assessed as grade D. The distribution of histologic findings was not significantly skewed within the respective grading categories (only 37.5% of grade A endometria, 63.5% of grade B endometria and 66.5% of grade C endometria were in phase with the menses). The in-phase and out-of-phase endometria did not display significant differences in endometrial thickness (8.8 +/- 0.29 mm vs. 9.13 +/- 0.4mm). Neither the sonographically measured endometrial thickness nor the echo pattern correlated with the histologic findings, suggesting that ultrasonography is inadequate for drawing reliable conclusions about endometrial receptivity in an IVF program.

Tetraploidy and partial endoreduplication in a tripronuclear zygote obtained after intracytoplasmic sperm injection

OBJECTIVE To describe a peculiar combination of cytogenetic abnormalities in a tripronuclear zygote obtained after intracytoplasmic sperm injection (ICSI). DESIGN Case report. SETTING A university hospital. PATIENT(S) A couple with a 4-year history of primary infertility. Intracytoplasmic sperm injection was performed because of male factor infertility (oligoteratozoospermia). INTERVENTION(S) Ultrasound-guided transvaginal follicular aspiration. MAIN OUTCOME MEASURE(S) Chromosomal karyotype of a tripronuclear one-cell zygote. RESULT(S) Unexpectedly, a tetraploid [92,XXYY, end3, -18, end18] chromosome complement was found, indicating injection of a diploid spermatozoon carrying two Y chromosomes. The parental origin of the other abnormalities could not be determined. The missing chromosomes may be attributed either to a hypodiploid [44,YY,-18,-18] sperm cell or to a hypohaploid [22,X,-18] oocyte. The exact tetraploid count was restored by endoreduplication of two chromosomes. This event could have occurred in one and the same or in two different pronuclei. CONCLUSION(S) Cytogenetic analysis of multipronuclear zygotes appears useful for assessing the incidence of chromosomal abnormalities at the earliest stage of conception. In addition to other methods, it also may contribute to evaluation of the transmission of aberrations by spermatozoa from infertile men.

CYTOGENETICS OF HUMAN SPERMATOZOA : THE FREQUENCY OF VARIOUS CHROMOSOME ABERRATIONS AND THEIR RELATIONSHIP TO CLINICAL AND BIOLOGICAL PARAMETERS

Human sperm chromosomes can be visualized after their fusion with zona-free hamster eggs. We analyzed a total of 867 metaphases from 33 men that had been classified into three different groups. We present a detailed summary of the kind and frequency of chromosome aberrations we encountered. A comparison of our results with previous studies demonstrates considerable variations in numerical and structural chromosome anomalies in spermatozoa from normal, healthy men. The possible reasons for this are discussed. We reviewed some factors which are supposed to correlate with the occurrence of sperm chromosome aberrations; the effect of paternal age is still uncertain.

Is there any correlation between follicular fluid hormone concentrations, fertilizability, and cytogenetic analysis of human oocytes recovered for in vitro fertilization?

Constituents of FF have been suspected to influence fertilizability and cytogenetic constitution of human oocytes. We therefore analyzed the FF concentrations of E2, P, T, and PRL for 114 oocytes recovered for IVF. Forty-six of these oocytes were fertilized and transferred to the maternal uterus. Among the unfertilized gametes, 27 were not analyzable, 30 were normal haploid, and 11 were classified as abnormal. There was no significant difference between fertilized and unfertilized oocytes for FF concentrations of E2, P, T, and PRL and for the E2:P ratios. Similarly, we detected no significant difference between normal and abnormal oocytes for these parameters.