Norman Ginsberg

Prevention of Age-Related Aneuploidies by Polar Body Testing of Oocytes

Purpose:We previously demonstrated that aneuploidy-free oocytes may be preselected by testing the first and second polar bodies removed from oocytes following their maturation and fertilization. The present paper describes the results of the application of the method in 659 in vitro fertilization cycles from patients of advanced maternal age.Methods:Using micromanipulation techniques, 3943 oocytes were tested by polar body sampling and fluorescent in situ hybridization analysis using specific probes for chromosomes 13, 18, and 21.Results:Fluorescent in situ hybridization results were available for 3217 (81.6%) of 3943 oocytes studied, of which 1388 (43.1%) had aneuploidies; 35.7% of the aneuploidies were of first meiotic division origin, and 26.1% of second meiotic division origin. Most errors in the first meiotic division were represented by chromatid malsegregation. The transfer of embryos deriving from 1558 of 1829 aneuploidy-free oocytes in 614 treatment cycles resulted in 131 clinical pregnancies and 88 healthy children born after confirmation of the polar body diagnosis.Conclusions:Polar body testing of oocytes provides an accurate and reliable approach for prevention of age-related aneuploidies in in vitro fertilization patients of advanced maternal age.

Preimplantation Diagnosis of Common Aneuploidies by the First- and Second-Polar Body FISH Analysis

Purpose:A low pregnancy rate in in vitro fertilization (IVF) patients of advanced maternal age may be caused by aneuploidies originating from non disjunction in the first or second meiotic divisions. We introduced genetic testing of oocytes by sampling and fluorescent in situ hybridization (FISH) analysis of the first and second polar bodies, to avoid fertilization and transfer of aneuploid oocytes in IVF patients of advanced maternal age.Methods:Three hundred and sixty-three IVF patients 34 years and older participated in the study. Using micromanipulation procedures, the first and second polar bodies were removed following their extrusion from the oocytes and studied by FISH, using probes specific for chromosomes 13, 18, and 21 to detect oocytes with common aneuploidies.Results:Of a total of 538 IVF cycles, 3250 oocytes were available for FISH analysis, with conclusive FISH results in 2742 oocytes (84.3%). As many as 1102 (40%) of oocytes were predicted to be aneuploid and not transferred. Of 1640 embryos predicted to be normal, 1145 were transferred in 467 treatment cycles, resulting in 107 pregnancies (23%), from which 67 healthy children have been born, 32 pregnancies spontaneously aborted, and 15pregnancies are ongoing after being confirmed normal by prenatal diagnosis.Conclusions:Preimplantation diagnosis by first- and second-polar body FISH analysis allows us to avoid the age-related risk of common aneuploidies in IVF patients of advanced maternal age.

Polar body diagnosis of common aneuploidies by FISH

AbstractPurpose: The purpose of this work was to investigate the reliability and accuracy of polar body analysis for preimplantation diagnosis of common aneuploidies in IVF patients of advanced maternal age. Design: We have previously introduced polar body analysis as an approach for nondestractive evaluation of the genotype of human oocytes. The method has recently been applied in a clinical trial involving 45 infertile patients, demonstrating the feasibility of preconception diagnosis of common aneuploidies by fluorescent in situ hybridization (FISH). The present paper describes the experience of polar body diagnosis in 135 IVF patients (161 cycles) of advanced maternal age. Results: FISH results of the first and/or second polar bodies were available in 648 (72.4%) of 895 biopsied oocytes subjected to FISH analysis. Of 648 oocytes with FISH results, 208 demonstrated chromosomal abnormalities. Of 440 oocytes predicted to be free from monosomy or trisomy of chromosomes X, 18, and/or 13/21, 314 were normally fertilized, cleaved, and transferred in 122 treatment cycles, resulting in 6 healthy deliveries and 12 ongoing pregnancies following confirmation of the polar body diagnosis by CVS or amniocentesis. Conclusion: The method may be useful for detection of oocytes with common chromosomal trisomies in IVF patients of advanced maternal age.

Analyses of 95 first-trimester spontaneous abortions by chorionic villus sampling and karyotype.

PurposeOur purpose was to determine the incidence of chromosomal aneuploidy in first-trimester pregnancy losses using chorionic villus sampling (CVS).Study DesignAll patients presenting for CVS with no fetal cardiac activity were offered CVS.ResultsCytogenetic results were completed in 95 of 96 cases (99%). Eighty-three percent of the karyotypes were aneuploid. The 16 euploid fetuses had no excess of females.ConclusionCVS is the most reliable method of determining the karyotype of spontaneously aborted fetuses. The incidence of aneuploidy is much greater than in previous reports that analyzed passed products of conception. CVS should be offered to women who present with first-trimester spontaneous abortions.

Ultrasonographic detection of the second-trimester fetus with trisomy 18 and trisomy 21

Biparietal diameter/femur length ratio and nuchal thickness were found to be sensitive indicators for the prenatal detection of trisomy 18 and trisomy 21. A biparietal diameter/femur length ratio greater than 1.5 SD above the control mean correctly identified 5 of 11 (46%) fetuses with trisomy 21 and 3 of 4 (75%) fetuses with trisomy 18. Nuchal thickening (6 mm or more) correctly identified 5 of 12 (41%) fetuses with trisomy 21 and 2 of 4 (50%) fetuses with trisomy 18. The sensitivity and specificity of the biparietal diameter/femur length ratio in detecting either aneuploidy was 53% and 93%, respectively, whereas a thickened nuchal fold had a sensitivity of 44% and a specificity of 100%. The combined use of the two ultrasonographic measurements had an overall sensitivity of 81% and a specificity of 93%. Prospective ascertainment of these two trisomies appears warranted in low-risk populations.

Fusion as the etiology of chimerism in monochorionic dizygotic twins.

In a dizygotic pregnancy within monochorionic placenta, findings consistent with chimerism were detected. Monochorionicity was confirmed by a combination of ultrasound, histological evaluation and DNA technology. Etiologic hypotheses are offered to explain this rare circumstance.

Founder Fukutin mutation causes Walker–Warburg syndrome in four Ashkenazi Jewish families

Walker‐Warburg syndrome (WWS) is a genetically heterogeneous congenital muscular dystrophy caused by abnormal glycosylation of α‐dystroglycan (α‐DG) that is associated with brain malformations and eye anomalies. The Fukutin (FKTN) gene, which causes autosomal recessively inherited WWS is most often associated with Fukuyama congenital muscular dystrophy in Japan. We describe the clinical features of four nonconsanguinous Ashkenazi Jewish families with WWS and identify the underlying genetic basis for WWS.

Detection of translocations involving the Y‐chromosome in prospective prenatal screening of common chromosomal aneuploidies by FISH

In the application of the fluorescence in situ hybridization (FISH) technique for prospective prenatal screening of common aneuploidies involving the autosomes 13, 18, and 21, and sex chromosomes, six cases of inconsistency between the results of FISH analysis and the results of karyotyping of cultured amniocytes have been observed, including two cases of translocation involving the Y‐chromosome and chromosome 15 in a total of 904 cases of amniocentesis studied. In one case, the translocation was of maternal origin, and in the other, of paternal origin. In both cases, the couples decided to continue the pregnancy and normal babies were delivered. The data show the usefulness of applying the FISH technique in prospective prenatal screening of common trisomies for the possible detection of rare chromosome rearrangements involving the Y‐chromosome.

Prenatal diagnosis of 46,XX male fetuses ☆ ☆☆

Ultrasonography can accurately determine phenotypic sex differences from those of the genetic sex. Two cases were identified; they were the result of a translocation of the SRY gene from the Y chromosome to the X chromosome during meiosis. An ultrasonographic difference may represent an otherwise unsuspected genetic abnormality.

Microdeletions/duplications involving TBX1 gene in fetuses with conotruncal heart defects which are negative for 22q11.2 deletion on fluorescence in-situ hybridization

Conotruncal heart defects (CTD) are associated with del22q11.2 syndrome, which is often diagnosed by fluorescence in‐situ hybridization (FISH). However, in those negative for del22q11.2 on FISH, the etiology is usually obscure. We aimed to use high‐resolution array comparative genomic hybridization (array CGH) to clarify the underlying genetic causes in these cases.