Simon Kipersztok

Validity of self-report for fractures among a multiethnic cohort of postmenopausal women: results from the Women's Health Initiative observational study and clinical trials.

Objective:The purpose of this study is to examine the validity of, and factors associated with, the accuracy of self-report (participant-report and proxy-report) for fractures. Design:Study participants were from the Womens Health Initiative Clinical Trial and Observational Study cohorts. All women were postmenopausal; populations included American Indian, Asian/Pacific Islander, black, Hispanic, and non-Hispanic white. The average length of follow-up was 4.3 years. Self-reported fractures were adjudicated by reviewing medical records. The first adjudicated self-report of fractures for each participant was included in the analysis (n = 6,652). Results:We found substantial variations in validity of self-report by the fracture site. Agreements between self-reports for single-site fractures and medical records were high for hip (78%) and forearm/wrist (81%) but relatively lower for clinical spine fractures (51%). The average confirmation rate for all single-site fractures was 71%. Misidentification of fracture sites by participants or proxy-reporters seemed to be a cause of unconfirmed self-reports. Higher confirmation rates were observed in participant-reports than in proxy-reports. Results of the multivariate analysis indicated that multiple factors, such as ethnicity, a history of osteoporosis or fractures, body mass index, years since menopause, smoking status, and number of falls in the past year were significantly (P < 0.05) related to the validity of self-report. Conclusion:The validity of self-reports for fracture varies by fracture sites and many other factors. The assessed validity in this study is likely conservative because some of the unconfirmed self-reports may be due to poor medical record systems. The validity of self-reports for hip and forearm/wrist fractures is high in this study, supporting their use in epidemiological studies among postmenopausal women.

Contribution of male age to outcomes in assisted reproductive technologies

OBJECTIVE To evaluate the relationship between male age and pregnancy outcome in donor oocyte assisted reproductive technology cycles. DESIGN Retrospective cohort. SETTING Private IVF center. PATIENT(S) A total of 1,392 donor cycles from 1,083 female recipients and their male partners. INTERVENTION(S) Oocyte donor cycles. MAIN OUTCOME MEASURE(S) Live birth. RESULT(S) Increasing male age was associated with semen parameters including volume and motility; however, male age was not observed to have a statistically significant association with likelihood of live birth in donor cycles after adjustment for female recipient age. CONCLUSION(S) When treatment cycle number and female recipient age were taken into account, male age had no significant association with pregnancy outcomes in assisted reproductive technology donor cycles in this study population.

Results on single cell PCR for Huntington's gene and WAVE™ product analysis for preimplantation genetic diagnosis

Triple repeat base pair amplification is the basis for a number of prevalent genetic diseases such as Huntingtons, Fragile X, Myotonic Dystrophy and others. We have chosen to investigate the use of PCR to amplify a portion of the Huntingtons gene in single cells in order to develop a clinical test system for preimplantation genetic diagnosis (PGD). Amplification of CAG triple repeat sequences poses difficulties due to resistance of GC melting for amplification. Special PCR modifications are necessary to carry out the amplification of GC rich areas found in most triple base pair expansions. We have used a modified polymerase chain reaction (PCR) protocol to amplify the expanded repeat sequence of the Huntingtons gene with satisfactory efficiency. Detection of the amplified expanded CAG repeats is shown to be possible using both agarose gel electrophoresis and high definition denaturing high pressure liquid (DHPLC) chromatography. The incidence of allele dropout (ADO) is documented.

Primer System for Single Cell Detection of Double Mutation for Tay-Sachs Disease

AbstractPurpose: Nearly 100% of infantile Tay-Sachs disease isproduced by two mutations occurring in the alpha chain ofthe lysosomal enzyme beta-N-acetylhexosaminidase (HEXA)in the Ashkenazi Jewish population. Although others havedescribed primer systems used to amplify both sitessimultaneously, few discuss the allele dropout problems inherent inthis test. Our goal was to construct a more robust testenabling stronger signal generation for single cellpreimplantation genetic diagnosis and to investigate theoccurrence of allele dropout. Methods: New nested primers were designed to optimizedetection of both major Tay-Sachs mutations. Four hundredfifty-seven single cells, including normal cells and thosecarrying mutations of either the 4bp insertion exon 11 orsplice-site intron 12 defects, were used to screen a newprimer system. Results: Based on PCR amplified product analysis, totalefficiency of amplification was 85.3%, (390/457). The alleledropout rate for the 4bp insertion mutation in exon 11 andsplice-site mutation in intron 12 was 4.8% and 5.8%,respectively. Conclusons: Multiple mutation detection and analysiswithin the Tay-Sachs disease gene (HEXA) is possible usingsingle cells for clinical preimplantation genetic diagnosis.Alternative PCR primers and conditions offer variousmethods for developing systems compatible to specificprogram requirements.

Validity of a rapid assay for antisperm antibodies in semen

OBJECTIVE To determine the validity of a rapid assay for antisperm antibodies in semen. DESIGN Prospective comparison of the results of standard and rapid antisperm antibody assays performed simultaneously. SETTING Tertiary care infertility center. PATIENT(S) Couples who presented for infertility evaluation. INTERVENTION(S) Semen analysis and measurement of antisperm antibodies in semen using a standard and a rapid immunobead binding test (IBT). MAIN OUTCOME MEASURE(S) [1] Comparison of sperm parameters between semen-containing antisperm antibodies and semen free of antisperm antibodies. [2] Validation of the rapid test by calculation of sensitivity, specificity, positive and negative predictive values of the rapid assay using the standard assay as a gold standard. [3] Cost comparison of the standard and rapid test. RESULT(S) [1] Nine semen specimens with antisperm antibodies had a significantly lower sperm concentration, motility, and total motile fraction compared to 44 specimens without antisperm antibodies. Also, specimens with antisperm antibodies had a significantly higher percentage of vibratory sperm and percent of bound antisperm antibodies. The strict morphology, liquefaction time, semen volume, and white blood cell concentration were no different between the two groups. [2] Using a threshold of > or =12% of bound antisperm antibodies in the rapid assay, the sensitivity, specificity, positive and negative predictive values of the test are 100% when correlated with a threshold of > or =20% in the standard assay. Increasing the threshold in the standard assay decreases the specificity and positive predictive value of the rapid assay but not the sensitivity and the negative predictive value. [3] The cost of the rapid assay was 16% that of the standard test and its performance took 20% of the time it took to set and perform the standard test. CONCLUSION(S) A rapid test for antisperm antibodies is valid, reliable, and more cost and labor effective than a standard IBT.

Pregnancy rates in varying age groups after in vitro fertilization: a comparison of follitropin alfa (Gonal F) and follitropin beta (Follistim).

OBJECTIVE Our purpose was to assess the efficacy of two recombinant follicle-stimulating hormones, follitropin beta (Follistim, Organon, West Orange, NJ) and follitropin alfa (Gonal F, Serono, Norwell, Mass) on pregnancy rates in varying age groups of women undergoing in vitro fertilization (IVF). STUDY DESIGN Three hundred sixty-five IVF cycles were retrospectively compared, 233 by use of follitropin beta and 132 by use of follitropin alfa, both after gonadotropin-releasing hormone agonist down-regulation. Assignment to each medication was indiscriminate. The primary outcome measured was pregnancy evidenced by fetal heartbeat on ultrasonography. Secondary outcomes included days of stimulation, ampules per patient cycle, estradiol level on the day of human chorionic gonadotropin administration, total follicles present on the day of human chorionic gonadotropin administration, follicles greater than 14 mm, oocytes retrieved, mature eggs, fertilization rate, and embryos transferred. Outcomes were stratified by age, including women less than 36 years old, 36 to 39 years old, and more than 39 years old. RESULTS There was no significant difference between follitropin beta and follitropin alfa in either the primary or secondary outcomes, although the pregnancy rate was significantly decreased with advancing age. CONCLUSION Success rates are similar, when stratified by age, in women undergoing IVF with either follitropin beta or follitropin alfa.

Simultaneous Single-Cell Detection of Two Mutations for Cystic Fibrosis

AbstractPurpose: A single-cell diagnosis procedure using polymerase chain reaction (PCR) technology was developed to simultaneously detect two cystic fibrosis (CF) mutations (DF-508, W1282X). Methods: Human viable, arrested, and nonviable embryos and immature, and nonfertilized oocytes donated by our patients were used to detect apoptosis by Tunel labeling, annexin staining, and single-cell reverse transcriptase–polymerase chain reaction (RT-PCR). Results: Using cells carrying the DF-508 mutation, the PCR signal efficiency for the affected homozygous, normal homozygous, and carrier heterozygote cell populations were 91%, 81%, and 92%, respectively. The total combined PCR efficiency was 87.7% and the ADO rate was 5.7%. For W1282X carrier heterozygote cells, the PCR signal efficiency was 82.0% and the ADO rate was 8.7%. Conclusions: Methods have been developed to detect two common mutations simultaneously for CF in single-cell assays. The high signal efficiencies and low ADO rates obtained in these tests allow those embryos from couples wishing to avert the transmission of this serious genetic disease to their offspring to be screened by preimplantation genetic diagnosis.

Twelve-well culture plate for the efficient collection and culture of human oocytes and embryos

AbstractObjective: Our goal was to assess a 12-well oocyte collection and embryo culture plate for use in the IVF laboratory. Design: This was a prospective nonrandomized study. Setting: The setting was a university in vitro fertilization program. Patients: Eighty-four consecutive infertility couples presenting for IVF were studied. Main Outcome Measures: The main outcome measure was pregnancy (delivered). Results: A 34% delivered pregnancy rate per retrieval was attained using the 12-well collection and culture plate without the use of expensive culture media and special serum supplementation. Conclusions: The use of a 12-well plate for oocyte collection and embryo culture provides a simple, economical, efficient, and effective means of producing human embryos during in vitro fertilization. This system is capable of supporting high rates of ongoing and delivered pregnancies.