Steen Smidt-Jensen

The role of DNA strand breaks in human spermatozoa used for IVF and ICSI

Background. The objective of this study was to determine the incidence of spermatozoa with DNA strand breaks in four clinically different groups of infertile couples, and to correlate DNA damage with other semen analysis parameters, as well as fertilization rates and IVF outcome.

Randomised comparison of amniocentesis and transabdominal and transcervical chorionic villus sampling

We have compared three methods of prenatal diagnosis in two large obstetric centres in Denmark. Women were randomly assigned transabdominal (TA) chorionic villus sampling (CVS), transcervical (TC) CVS, or second-trimester amniocentesis (AC); women at high genetic risk were randomised between the two CVS groups only. Analysis of 45 epidemiological variables showed the three procedure groups to be similar at enrollment. All women were followed up until completion of pregnancy. Among 3079 women at low genetic risk total fetal loss rates were 10.9% for TC CVS, 6.3% for TA CVS, and 6.4% for AC (p < 0.001). More women had bleeding after the procedure in the CVS groups (p < 0.001), whereas more amniotic fluid leakage (p < 0.001) was reported after AC. No uterine infections occurred in any group. No case of oromandibular-limb abnormality was seen in the CVS groups, but 1 child in the AC group had aplasia of the right hand. The two CVS approaches were compared among 2882 women at low and high genetic risk who were found to have cytogenetically normal fetuses. Rates of unintentional loss after the procedure were 7.7% for TC CVS and 3.7% for TA CVS (p < 0.001; 95% Cl of difference 2.3-5.8%). At baseline ultrasound scanning after establishment of optimum sampling conditions, more TC than TA procedures (p < 0.001) were judged not to be feasible. We found that TA CVS allows better access to the placental site than TC sampling, is an easier skill to acquire, and has the potential that more villi can be aspirated when needed. The risk of fetal loss is similar after TA CVS and AC. However, losses after AC are at a later stage and are therefore more distressing. TA procedures remain the first choice for prenatal diagnosis. Since, in our hands, TC sampling carries a greater risk to the fetus, we have abandoned TC CVS in our two study centres.

Apoptosis in human cumulus cells in relation to zona pellucida thickness variation, maturation stage, and cleavage of the corresponding oocyte after intracytoplasmic sperm injection

OBJECTIVE To assess the degree of apoptosis in the cumulus cells and the variation of the zona pellucida and the maturity and fertilization of the corresponding oocyte. DESIGN Retrospective study. SETTING Private fertility clinic. PATIENT(S) Fifty couples undergoing ICSI. INTERVENTION(S) ICSI. MAIN OUTCOME MEASURE(S) Correlation between apoptosis in the cumulus cells and the zona pellucida thickness variation, maturation stage, fertilization rate, and embryo score. RESULT(S) This study demonstrated no correlation between apoptosis in cumulus cells and the thickness and variation of the zona pellucida in oocytes and embryos. The incidence of apoptosis was significantly higher in cumulus cells from empty zona pellucidas and germinal vesicle stage and metaphase I oocytes compared with metaphase II oocytes. Non-fertilized metaphase II oocytes showed significantly higher incidence of apoptosis compared with fertilized metaphase II oocytes. There was a correlation between embryo score and the zona pellucida thickness variation. CONCLUSION(S) Apoptosis in cumulus cells had no impact on the zona pellucida thickness and variation in oocytes and embryos. The zona pellucida thickness variation was positively correlated to good embryo score. A higher degree of apoptosis was seen in cumulus cells from immature oocytes compared with mature oocytes. Furthermore, apoptosis in cumulus cells impaired the fertilization rate of metaphase II oocytes after ICSI.

DNA strand breaks in human spermatozoa: Correlation with fertilization in vitro in oligozoospermic men and in men with unexplained infertility

Background. The purpose of this study was to examine the correlation between DNA strand breaks in human spermatozoa and semen quality, fertilization rate and IVF outcome.

Fast track vaginal surgery

Objective. Our aim was to describe the need for postoperative hospitalization after vaginal surgery for utero‐vaginal prolapse with well‐defined charts for postoperative care.

Apoptosis in human cumulus cells in relation to maturation stage and cleavage of the corresponding oocyte

Background. The purpose of this study was to determine the incidence of apoptosis in cumulus cells, and correlate these findings with the maturation stage, fertilization rate and embryo score of the corresponding oocyte, in couples undergoing ICSI due to a male factor.

Morphology of spermatozoa used in IVF and ICSI from oligozoospermic men

The purpose of this study was to determine morphology in human spermatozoa from men with oligozoospermia and correlate with the fertilization rate, embryo score and pregnancy rate after IVF and intracytoplasmic sperm injection (ICSI) respectively. The study group consisted of 125 couples where the male partner suffered from oligozoospermia. Fifty of these had IVF (group A). Seventy-five couples in whom ICSI had been performed made up group B. Sperm samples were assessed according to the WHO manual. For each male, morphology of spermatozoa was judged according to Krügers strict criteria, WHO criteria and the teratozoospermia index (TZI). Oocyte monitoring was carried out according to a long down-regulation protocol using gonadotrophin-releasing hormone, recombinant FSH and human chorionic gonadotrophin. Embryos were transferred on day 2 after fertilization, with a maximum of three embryos. This study demonstrated no correlation between any of the morphological assessments of spermatozoa and the fertilization rate, embryo score and pregnancy rate, either after IVF or ICSI. Morphology in human spermatozoa according to Krügers strict criteria, WHO criteria and the TZI had no predictive value for the outcome after either IVF or ICSI.

Fetal Cells in Maternal Blood: A Comparison of Methods for Cell Isolation and Identification

Objective: A variety of methods have been used to select and identify fetal cells from maternal blood. In this study, a commonly used 3-step selection method is compared with selection directly from whole blood. Identification of fetal origin by XY FISH of male cells was also evaluated. Methods: Maternal blood was drawn either before invasive chorion villus sampling (pre-CVS) or after (post-CVS) from women carrying a male fetus. Fetal cells were isolated either by density gradient centrifugation succeeded by CD45/CD14 depletion and CD71-positive selection from CD45/CD14-negative cells, or by CD71-positive selection directly from whole blood. The true origin of fetal cells recovered by the two methods was established by two rounds of XY chromosome FISH in reverse colors, in some instances combined with anti-zeta (ζ) or anti-ζ/anti-gamma (γ) antibody staining. Results: In blood samples taken post-CVS and enriched by CD71 selection directly from whole blood, fetal cells were identified with a frequency that was almost four orders of magnitude higher than in post-CVS samples enriched by the 3-step method. In blood samples taken pre-CVS and enriched by the 3-step procedure, no fetal cells were identified by reverse color FISH in 371 ml of blood. In similar samples enriched by CD71 selection on whole blood, two fetal cells were identified in 27 ml of blood. Rehybridization with X and Y chromosome probes with reverse colors was necessary to exclude false Y chromosome signals. Not all fetal cells identified by the presence of a true Y chromosome signal stained with anti-ζ antibody. Conclusions: Selection of fetal NRBCs from maternal blood by CD71-positive selection directly from whole blood is superior to density gradient centrifugation succeeded by CD45/CD14 depletion and CD71 selection of CD45/CD14-negative cells. Combining two markers for fetal origin is recommended for unambiguously identifying a cell as fetal.

A case of first trimester chromosomal mosaicism confined to the cultivation of the gestational products

The advantages of the emergence and developmcnt of chorionic villi sampling (CVS) for early prenatal diagnosis are evident, but there are a host of new diagnostic problems caused by the use of extraembryonic tissues. We report a case in which 45X/46XY mosaicism was diagnosed by cultivation of chorionic villi and fetal cells. Direct chromosomal preparations of chorionic villi failed to diagnose the abnormality.

The safety of chorionic villus sampling. A synthesis of the literature.

Altogether 10 reports on the safety of chorionic villus sampling, either by the transcervical (TC) or the transabdominal (TA) approach, were reviewed and combined with our own data. After discussion of how unintended fetal loss rates are best estimated, the excess total fetal loss after TC and TA compared with amniocentesis were estimated to be 1.70% (+/- 0.65%) and practically zero (+/- 1.0%), respectively (standard errors in parentheses). The absolute risk of unintended loss after TC is +2.7% (+/- 0.7%) and after TA 1.0% (+/- 1.0%). These estimates are still too uncertain to allow precise weighting of benefits and human costs. A uniform style of reporting studies in this area is proposed.