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B Rosenbusch

Publications and source records attributed to B Rosenbusch.

At least 19 recordsLinked to original sources
PubMed1998

Maturation of a binuclear oocyte from the germinal vesicle stage to metaphase II: formation of two polar bodies and two haploid chromosome sets.

We report on a binuclear human oocyte that underwent maturation in vitro from germinal vesicle stage to metaphase II. Following extrusion of two polar bodies, the oocyte was processed for cytogenetic analysis which revealed two separate haploid chromosome sets accompanied by the corresponding polar body chromatin. Tentatively established karyotypes were 23,X and 23,X,ace, respectively. This condition could have resulted in a tripronuclear digynic zygote after monospermic fertilization.

Adult↗
PubMed1998

Chromosomal analysis of multipronuclear zygotes obtained after partial zona dissection of the oocytes.

We have attempted to analyse the chromosome constitution of multipronuclear 1-cell zygotes obtained after partial zona dissection (PZD) of the oocytes. Six cells with three pronuclei could not be evaluated whereas another one was characterized by the presence of a normal haploid and two uninterpretable metaphases. Complete karyotypes were established for 21 tripronuclear cells, taking the varying arrangement of the chromosome sets into consideration. Of the zygotes, 10 showed three separated haploid metaphases (distribution pattern n/n/n), eight zygotes had one haploid and one diploid chromosome set (n/2n) and in three cells the individual sets were not distinguishable (3n). The sex chromosome ratio XXX:XXY:XYY was 7:9:5. Chromosome abnormalities were found in eight of the completely or partially analysable tripronuclear zygotes (36.4%) and included numerical (4 cells), structural (2 cells) as well as combinations of numerical and structural alterations (2 cells). Three out of 11 zygotes with four pronuclei could not be evaluated at all. In three cases, only two chromosome sets were analysable and another cell displayed one uninterpretable set. Three out of eight completely or partially analysable zygotes with four pronuclei (37.5%) had chromosomal abnormalities. Excluding the four cells with one or two uninterpretable metaphases, the sex chromosome distribution XXXX:XXXY:XXYY:XYYY in the zygotes with four pronuclei was 0:1:1:2. Compared with previously analysed multipronuclear zygotes obtained after conventional in-vitro fertilization (IVF), the rate of aberrant zygotes as well as the incidence of aberrant (male + female) chromosome sets were not significantly changed after PZD.

Adult↗
PubMed1997

The chromosomal constitution of multipronuclear zygotes resulting from in-vitro fertilization.

We have attempted to analyse the chromosome constitution of 77 multipronuclear uncleaved zygotes obtained from our in-vitro fertilization programme. Complete karyotypes could be established for 51 tripronuclear cells and eight zygotes with four pronuclei. When compiling the results, the varying arrangement of the chromosome sets was taken into consideration. Eighteen tripronuclear zygotes showed three separate haploid metaphases (distribution pattern n/n/n), 16 cells had one haploid and one diploid chromosome set (n/2n), and in 15 zygotes the individual sets were not distinguishable (3n). Two zygotes were in fact tetraploid, the distribution of metaphases on the slide being n/3n and n/n/2n, respectively. In tripronuclear zygotes the sex chromosome ratio XXX:XXY:XYY was 14:16:18, excluding the two tetraploid cells and one zygote with a 23,X/23,X/22,-C or -Y karyotype. Chromosome abnormalities were found in 16 zygotes (31.4%) and included numerical (six cells), structural (four cells) as well as combinations of numerical and structural alterations (six cells). Four of the zygotes with four pronuclei (50%) had numerical and/or structural chromosome aberrations. Excluding two cells with one uninterpretable metaphase and a 22,-C or -Y karyotype, respectively, the sex chromosome distribution XXXX:XXXY:XXYY:XYYY was 1:1:2:1 in zygotes with four pronuclei. Another zygote was found to be pentaploid after fixation. These results suggest that analysis of multipronuclear zygotes yields valuable information about cytogenetic abnormalities occurring at the earliest stage of conception.

Adult↗
PubMed1997

Triploidy caused by endoreduplication in a human zygote obtained after in-vitro fertilization.

Cytogenetic analysis of a presumably tripronuclear zygote revealed that triploidy was caused by an endoreduplicated 46,XX complement. The remaining chromosomes yielded a hyperhaploid karyotype of 28,Y, +2, +3C, +D. The origin of this chromosomal composition is obscure. Besides endoreduplication in a normal 23,X oocyte pronucleus and fertilization by a normal 23,Y spermatozoon, an additional female pronucleus might have been formed due to an irregular chromosome distribution during second meiotic division. On the other hand, penetration by a hyperhaploid spermatozoon cannot be excluded with certainty.

Adult↗
PubMed1996

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 Women's 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.

Adult↗
PubMed1995

Predicting the success of in vitro fertilization: conventional semen analysis compared to the hamster ova penetration test.

In vitro fertilization has been advocated as a possible treatment for male factor infertility. Using data collected on 154 men, conventional semen analysis and the hamster ova penetration test are compared with respect to their power in predicting the success of assisted conception. As performing the hamster test is more expensive than the semen analysis, we also investigate a combined strategy. It performs better than the discrimination based solely on the semen parameters, but does not lead to an improvement over the overall discriminatory power obtained from the hamster test alone.

Adult↗
PubMed1995

[Idiopathic habitual abortion: experiences with active immunotherapy].

The effect of lymphocyte transfusions on the stabilisation and successful outcome of pregnancy was investigated in 30 women with recurrent abortions presumably caused by immunologic factors. Lymphocyte concentrates were obtained from stored whole blood of the husbands and transferred by intravenous injection. Exclusion criteria for this study were the presence of antipaternal lymphocytotoxic antibodies in the patient's serum (cross match) or an advanced pregnancy (> 12 weeks). After transfusion of paternal lymphocytes 19 (63%) women had uncomplicated and successful pregnancies whereas 11 (37%) suffered again from early pregnancy loss. Two out of these 11 patients aborted twice. The probability of a successful treatment decreased as soon as lymphocytotoxic antibodies appeared in the patient's serum (p < or = 0.021). Women with a lower number of abortions before treatment carried their pregnancies to full term more frequently (p < or = 0.03). The number of transfusions, the period between start of treatment and conception as well as the distribution of age within the two groups had no significant effect on the outcome of therapy. A lack of lymphocytotoxic antibodies or their late appearance during treatment give a positive prognosis for the progress of pregnancy. Compared with a fertile control group, couples with habitual abortions showed no significantly increased HLA sharing. Immunisation with paternal lymphocytes in cases of presumably habitual abortions due to immunologic factors appears as an effective therapy with only few side effects. Large, randomised, double-blind, and placebo-controlled trials are needed before drawing conclusions.

Abortion, Habitual↗
PubMed1995

[Cytogenetic abnormalities in sperm: a risk factor for application of medical reproduction techniques?].

Microinsemination techniques are increasingly applied to achieve fertilisation of the oocyte in cases of male fertility disorders. Thus, spermatozoa can directly interact with the oolemma or the ooplasm, thereby obviating the need for penetration through cumulus oophorus and zona pellucida. However, problems associated with this particular feature arise in view of the following considerations: 1. The gametes from subfertile males are suspected to carry more cytogenetic anomalies in addition to an impaired sperm quality when compared to fertile men. 2. Cumulus and zona play an important role in the selection of morphologically abnormal and dysfunctional spermatozoa. 3. Consequently, bypassing natural sperm selection processes would lead to more abnormal embryos, abortions, and births of diseased or handicapped children. Spermatozoa can be analysed cytogenetically after fusion with zona-free hamster eggs or by fluorescence in situ hybridization (FISH). Both techniques did not confirm increased aberrations in gametes from subfertile men with a normal somatic karyotype. However, a definite conclusion cannot be drawn because of the limited data and restrictions concerning their interpretation. This also holds true for the evaluation of correlations indicating no significant relationship between the chromosomal constitution and sperm motility or morphology, respectively. Currently, the risk of malformation after microinsemination does not appear to be increased when compared to conventional IVF but the results do not yet allow a reliable statistical evaluation. In contrast, decreased morphological quality, impaired implantation, and early abortions have been reported for embryos produced by assisted fertilisation. This suggests that an effective natural selection of abnormal embryos will prevent a significant increase in malformations in the following generation.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromosome Aberrations↗
PubMed1995

[Smoking--a possible cause for failure in IVF/ET treatment?].

The aim of this study was to investigate the relation between cigarette smoking of women and the fertilisation and pregnancy rates obtained by in-vitro fertilisation treatment. 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. The study population consisted of 197 women (23-39 years old) who were divided into the groups 'non-smokers' (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. There were no significant differences in fertilisation and pregnancy rates between the different groups (p > 0.05). The E2 serum levels were significantly decreased in women who smoked, compared with the results obtained from non-smokers and passive smokers (p < 0.025). Overall, a strong negative correlation of the cotinine- and E2 levels was observed (r = -0.65). The results suggest that there is no clinically detectable impairment of fertilisation potential due to female smoking and that there is a greater influence on the outcome of in-vitro fertilisation by other factors.

Adult↗
PubMed1994

[The effect of different anesthetic procedures on hormone levels in women. Studies during an in vitro fertilization-embryo transfer (IVF-ET) program].

Different anaesthetic procedures that were used during an in vitro fertilisation and embryo transfer (IVF-ET) program have been analysed in order to determine their influence on plasma levels of estradiol, progesterone, prolactin, and beta-endorphin and results of IVF-ET. METHODS. Fifty-four patients awaiting transvaginal oocyte aspiration were randomised into three groups: (1) anaesthesia with ketamine as an induction agent and analgesic (n = 20); (2) general intubation anaesthesia using thiopentone for induction and enflurane for maintenance (n = 18); and (3) no anaesthesia (n = 16). Estradiol, progesterone, prolactin, and beta-endorphin were measured from day 3 to 14 referring to follicle aspiration. Differences between preoperative hormone levels and their intra- and postoperative peaks were analysed using the Kruskal-Wallis test (P < 0.03). The results were corrected using the Holms method (alpha = 0.05). RESULTS. No differences were observed in estradiol and progesterone levels (Figs. 1, 2). Prolactin levels were 1.4 times higher (P < 0.001) when ketamine was used and 2.2 times higher (P < 0.001) after short general anaesthesia than in the control group (Fig.3). Similar results were observed with respect to beta-endorphin: in comparison with the control group we found significant elevation by a factor of 2.1 when ketamine was used (P < 0.001). The discrepancy became even more marked with general anaesthesia: beta-endorphin was 3.9 times higher compared to the controls (P < 0.001) (Fig.4). Comparing the two groups who were given anaesthetics, prolactin and beta-endorphin levels were also significantly different (P < 0.001). The IVF procedure itself did not appear to be affected by different anaesthetic procedures during oocyte aspiration (Table 2). CONCLUSIONS. The increased prolactin and beta-endorphin plasma levels associated with ketamine and general anaesthesia reflect a significant alteration of the observed hormone levels. When anaesthesia is indicated, we try to avoid general intubation anaesthesia in favor of ketamine.

Adult↗
PubMed1994

In vitro kallikrein treatment of human spermatozoa: stimulation of sperm penetration into zona-free hamster eggs depends on the quality of initial semen parameters but not on constituents of the kallikrein-kinin system in seminal plasma.

We investigated the effect of an in vitro kallikrein treatment of human spermatozoa on their ability to penetrate zona-free hamster eggs. Sixty-four infertility patients were classified into four groups (normo-, oligo-, astheno-, and oligoasthenozoospermia) in order to find out which men would benefit most from this procedure. An aliquot of each ejaculate was diluted with Ham's F-10 medium supplemented with human serum albumin and kept at 22C for 1 hour. Another aliquot was incubated with kallikrein (5 KU/ml.) for one hour at 22C. These sperm suspensions were then washed twice, subjected to a swim-up preparation, and subsequently incubated at 37C for 2 hours. The mean penetration rates of kallikrein-treated samples were significantly higher than those of the corresponding control assays in all but the oligoasthenozoospermia group. Our results suggest a direct proteolytic action of kallikrein because the stimulation of penetration into hamster eggs was not correlated with the amount of constituents of the kallikrein-kinin system (kinins, kininogen, kininase) in seminal plasma.

Animals↗
PubMed1994

[Do preparation techniques induce ultrastructural sperm damage? An electron microscopic study].

We investigated the possibility of ultrastructural damage to human spermatozoa induced by different sperm preparation techniques. Ejaculates from 20 normozoospermic men were divided into equal aliquots and processed by glass wool filtration, Percoll density gradient centrifugation, and a simple two-step centrifugation procedure which served as a control. Ultrastructural damage was assessed by scanning electron microscopy. Compared with Percoll density gradient centrifugation, glass wool filtration yielded a significantly increased proportion of intact acrosomes (p < 0.05). However, both preparations gave significantly better results than control (p < 0.001 and p < 0.01 for glass wool filtration and Percoll centrifugation, respectively). The significance of the conspicuous bending of sperm tails after Percoll centrifugation is not yet known. In conclusion, both glass wool filtration and Percoll centrifugation are efficient techniques for the accumulation of spermatozoa with intact acrosomes. Because of the higher sperm recovery rate, glass wool filtration appears to be the more appropriate method.

Acrosome↗
PubMed1994

[Initial pregnancies after intravaginal transport and partial zona dissection of human ova].

We report on 72 couples with previous failure of fertilisation during several trials of conventional in-vitro fertilisation (IVF). In the present study part of the aspirated oocytes was subjected to partial zona dissection (PZD) before insemination, thereby achieving 7 pregnancies during 104 cycles. Normal in-vitro insemination of the remaining zona-intact oocytes was again unsuccessful. Since microinsemination techniques, in contrast to conventional IVF, are not yet being offered by many clinics, we developed the following new concept: hormonal stimulation, sonographic control of follicular growth, aspiration of oocytes, and sperm preparation were performed at the usual place (Munich) whereas PZD and control IVF were accomplished in another laboratory (Ulm). In order to transport the gametes between these two institutes, we modified the previously reported intravaginal culture (IVC) of inseminated oocytes by transferring oocytes and spermatozoa into separate plastic capsules. In our view, this procedure offers a suitable opportunity for infertile couples living too far away from a microinsemination centre to afford long daily journeys or even several overnight stays for superovulation treatment. The fertilisation and pregnancy rates in our study are comparable to those reported in the literature.

Adult↗
PubMed1994

[Effect of different follicle puncture methods on hormone homeostasis in the woman. Studies within the scope of an in vitro fertilization-embryo transfer program].

Different methods for follicular puncture which were used in an in vitro fertilization-embryo transfer program have been analyzed in order to determine their influence on plasma levels of estradiol, progesterone, prolactin and beta-endorphins. Thirty-eight patients awaiting oocyte aspiration were randomized into a laparoscopic (n = 20) and an ultrasound-guided oocyte aspiration group (n = 18). No differences were observed as far as estradiol and progesterone levels and embryo transfer rates were concerned. When using the laparoscopic technique, prolactin levels were found to be elevated 1.5 times in comparison with the ultrasound-guided aspiration procedure, which is highly significant (p < 0.001). Also, beta-endorphins peaked at a significantly higher level during laparoscopy (27.8 vs. 20.7 mol/l, p < 0.001). The enhanced prolactin and beta-endorphin levels reflect an increased stress response during laparoscopy. With respect to less trauma and shorter duration of the procedure, we now perform the sonographically guided transvaginal aspiration technique in our program almost exclusively.

Adult↗
PubMed1994

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.

Adult↗