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Biomedical subjects

M M Seibel

Publications and source records attributed to M M Seibel.

At least 19 recordsLinked to original sources

Preimplantation genetic diagnosis of chromosome balance in embryos from a patient with a balanced reciprocal translocation.

Duplications or deletions are present in a high percentage of the gametes produced by individuals carrying balanced translocations. Preimplantation genetic diagnosis was used to examine chromosome balance in embryos from a patient having a reciprocal translocation within the short arms of chromosomes 5 and 8 (46,XX,t(5;8)(p13;p23)). This woman has two sisters with the translocation unbalanced, resulting in a partial trisomy for chromosome 5 and partial monosomy for chromosome 8 (46,XX,-8, +der(8)t(5;8)(p13;p23)) with associated mental retardation and physical abnormalities. The patient and her husband desired to have children without the abnormal chromosome balance and wished to reduce the likelihood of spontaneous abortion or need for therapeutic abortion. Fluorescence in-situ hybridization (FISH) probes for the alpha-satellite region of chromosome 8 and for a region on the short arm of chromosome 5 (5p15.2) were tested initially on lymphocytes from the patient and her sisters. The hybridization signal for chromosome 5 was detected in the expected two copies for the patient and three copies for the sisters in 87% of the cells. Two hybridization signals for chromosome 8 were detected in 96% of the cells from all individuals. Additional probe testing was done using blastomeres from polyspermic embryos. The couple then proceeded with a stimulated in-vitro fertilization (IVF) cycle and biopsies were done on 13 embryos at the 7-10-cell stage using a method of zona drilling and fluid displacement. Diagnosis was possible on at least one blastomere for nine embryos. Three embryos had nuclei with three hybridization signals for chromosome 5, three had fewer than two signals for one or both chromosomes, one was mosaic, and two had two signals for each chromosome. The latter were transferred to the patient, but pregnancy was not achieved. The results demonstrate that preimplantation genetic diagnosis for patients with reciprocal translocations can be used to identify embryos having normal chromosome balance. The potential advantages and limitations of this approach are discussed.

Abortion, Spontaneous

Expression of Wilms' tumor suppressor gene (WT1) in term human trophoblast: regulation by cyclic adenosine 3',5'-monophosphate.

The Wilms' tumor suppressor gene (WT1), which is deleted in some Wilms' tumors, encodes a zinc finger transcription factor. We studied WT1 messenger ribonucleic acid (mRNA) in human term placenta and cytotrophoblasts differentiating into syncytiotrophoblasts in vitro by RT-PCR. The results suggest that WT1 mRNA is expressed in the trophoblasts in a cell-specific fashion. WT1 mRNA expression has been observed to decline remarkably in trophoblast cells after 72 h, when these cells are morphologically differentiated into multinucleated syncytiotrophoblasts. As it is well known that cAMP as a second messenger plays a significant role in cellular proliferation and differentiation of placental cells, we examined the effect of 8-bromo-cAMP on WT1 mRNA expression in undifferentiated cytotrophoblasts and differentiated syncytiotrophoblasts. We observed that cAMP enhanced WT1 mRNA expression in cytotrophoblasts, but remained ineffective in altering WT1 mRNA in syncytiotrophoblasts. In summary, the results of this investigation demonstrate that the WT1 gene is developmentally regulated during trophoblast differentiation. An involvement of the cAMP-mediated system in regulating the WT1 gene in the trophoblast is suggested.

Cell Differentiation

Preimplantation development of mouse and human embryos biopsied at cleavage stages using a modified displacement technique.

A modified embryo biopsy method was tested on four- and eight-cell stage mouse embryos and used on human embryos to obtain blastomeres for preimplantation genetic diagnosis. The biopsy method tested combines zona drilling and fluid displacement to force one or two cells through an opening in the zona pellucida of the cleavage-stage embryo. Rates of cell division and the percentage of mouse embryos forming blastocysts following biopsy at the eight-cell stage were not significantly different from those observed in unoperated control embryos. The percentage blastocyst formation was not significantly different in embryos biopsied at the four-cell stage and in control embryos, although cell division was significantly retarded following biopsy. 96% of the mouse blastomeres isolated at the eight-cell stage were recovered intact and 96% of those placed in culture underwent cell division. Survival and division of cells isolated at the four-cell stage were 92 and 84% respectively. Most of the cultured blastomeres cleaved several times and formed small trophoblast vesicles. Chromosomes were observed in 59% of blastomeres incubated in the presence of colcemid. In the initial use of this biopsy technique for human preimplantation genetic diagnosis, blastocyst formation was observed in 9 of 13 human embryos biopsied at the 7- to 10-cell stage. These findings support the use of this biopsy method as an alternative to aspiration techniques.

Animals

Transvaginal amniotic puncture for cytogenetic evaluation of missed abortions.

Transvaginal amniotic puncture (TAP) was performed on 20 consecutive missed abortions immediately prior to dilatation and evacuation and the cytogenetic results compared. The information received from products of conception (POC) and TAP was in concordance in only 5 of 20 (25%) cases. Tissue obtained from POC yielded cells in all instances. However, only 3 of 20 POC samples yielded findings other than normal female. In contrast, 92.8% of the conclusive diagnoses would have been achieved by TAP alone. These data strongly suggest that TAP is superior to POC for accurate cytogenetic assessment of missed abortion and should lead to a reevaluation of our current understanding and management of pregnancy loss.

Abortion, Missed

Parameters that predict success for natural cycle in vitro fertilization-embryo transfer.

OBJECTIVES: To analyze our natural cycle IVF-ET experience in order to identify parameters predictive for success. DESIGN: Retrospective study. SETTING: Private-based reproductive center. PATIENTS: All patients who presented for IVF-ET interested in unmedicated cycles. MAIN OUTCOME MEASURES: Routine monitoring for IVF-ET was performed. In addition, serum values for FSH, PRL, E2, and P were analyzed. RESULTS: Day 3 FSH values were not useful, whereas elevated day 3 LH values adversely correlated with outcome. Age was the single most important predictor of success. With two cycles, 87.5% of pregnancies occurred. CONCLUSION: With proper patient selection, natural cycle IVF-ET results can be substantially improved.

Adult

The developmental potential of mouse oocytes matured in serum-free culture conditions.

Studies were undertaken to identify serum-free conditions for the maturation of mouse oocytes in vitro. Oocytes were recovered from the antral follicles of juvenile mice 48 h after injection with gonadotrophin and allowed to resume meiosis in modified Ham's F-10 (mHF-10) medium unsupplemented or supplemented with bovine serum albumin (BSA), fetal calf serum, human pre-ovulatory serum, human follicular fluid or EDTA. They were inseminated 14-16 h later, scored for polar body extrusion after 4-6 h with spermatozoa, and transferred to protein-free mHF-10 for further development. In-vivo matured ova were inseminated and cultured in parallel as controls. Fertilization and development were scored as two cells 24 h after insemination and blastocysts 4 days following insemination respectively. Surprisingly, 41% of oocytes cultured in unsupplemented mHF-10 completed meiosis I, and of those, 50% fertilized; serum supplementation did not improve maturation or fertilization rates. Although the addition of human follicular fluid to the mHF-10 improved meiosis (69%) and fertilization (68% of eggs with polar bodies) to levels comparable with the in-vivo control eggs (79 and 66% respectively), BSA supplementation was equally beneficial. Blastocyst development varied, but within each maturation/fertilization group, the development from in-vitro matured eggs was comparable with embryos from in-vivo matured eggs. In addition, two out of eight 4-cell embryos from oocytes cultured in mHF-10 with BSA and EDTA gave rise to apparently normal pups following transfer to pseudopregnant recipients.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Semen leukocytes: friends or foes?

OBJECTIVE: To test the hypothesis that male reproductive tract leukocytes function in the elimination of abnormal spermatozoa from ejaculated semen. DESIGN: Semen specimens with > or = 2 x 10(6) nonspermatozoal cells/mL were examined for leukocytes and for mature sperm with ideal morphology. SETTING: Andrology laboratory of a Center of Assisted Reproductive Technology. RESULTS: Semen specimens with elevated concentrations of leukocytes contained a significantly higher frequency of sperm with ideal morphology than semen specimens with elevated numbers of immature germ cells and low numbers of leukocytes. CONCLUSIONS: The direct correlation between leukocyte density and sperm with ideal morphology supports the concept that sperm surveillance is a normal function of male reproductive tract leukocytes. Understanding such germ cell-leukocyte interactions may provide valuable new insights into immunologic control mechanisms in male reproductive tract tissues.

Adult

Toward reducing risks and costs of egg donation: a preliminary report.

OBJECTIVE: To use clomiphene citrate for ovarian stimulation of egg donors as a means of reducing the risk, discomfort, and inconvenience for egg donors and reducing costs for recipients. DESIGN: Prospective nonrandomized clinical study. SETTING: Private reproductive center. PATIENTS: Egg donation recipients and their anonymous donors. INTERVENTION: Clomiphene citrate was administered to egg donors rather than a GnRH agonist and gonadotropins before oocyte retrieval. MAIN OUTCOME MEASURE: Pregnancy. RESULTS: Eleven oocyte retrievals were performed and conception occurred in four cycles, three of which delivered. CONCLUSION: Clomiphene citrate appears to be a reasonable less expensive method of ovarian stimulation for egg donation in normospermic males and appears more acceptable to potential donors.

Adult