Anil Dubey

The correlation between follicular measurements, oocyte morphology, and fertilization rates in an in vitro fertilization program*

OBJECTIVE To explore the relationship between follicle size and the morphology of the oocyte-cumulus-corona complex with fertilization rates in stimulated cycles of IVF. DESIGN Retrospective comparison of measurements and observations of 2,429 oocytes from 215 patients undergoing 324 stimulated IVF cycles. SETTING A large hospital-based IVF program. MAIN OUTCOME MEASURES Individual follicles were measured by ultrasound before transvaginal aspiration and the size was recorded. The oocyte-cumulus-corona complex from each follicle was examined and classified. The oocytes were checked for evidence of fertilization 17 to 22 hours after insemination. RESULTS The fertilization rate of all oocytes regardless of morphological type revealed a positive linear correlation with increasing follicle diameter. The fertilization rates of type I oocytes was marginally higher than type II oocytes, controlling for follicle diameter; however, this difference did not achieve statistical significance. Oocytes from follicles with a mean diameter > or = 16 mm had significantly higher fertilization rates than did oocytes from follicles with a mean diameter < or = 14 mm. CONCLUSIONS Follicle size is a better predictor of fertilization than is morphological characterization of the oocyte-cumulus-corona complex in IVF.

Administration of progesterone before oocyte retrieval negatively affects the implantation rate

OBJECTIVE To compare the efficacy of two clinically accepted methods of progesterone supplementation during IVF. DESIGN Prospective randomized trial. SETTING A university-based IVF program. PATIENT(S) Three hundred fourteen stimulated IVF cycles between January 1993 and October 1994. INTERVENTION(S) Patients were assigned to one of two luteal phase progesterone regimens by a random permuted block design. In protocol A, 12.5 mg of IM progesterone was given 12 hours before oocyte retrieval; in protocol B, 25 mg of IM progesterone was given on the day of oocyte retrieval. MAIN OUTCOME MEASURE(S) Clinical pregnancy. RESULT(S) Patient demographic characteristics, including age, diagnosis, number of oocytes retrieved and fertilized, and number of embryos transferred, were not different between the two groups. There was no difference in the rate of cycle cancellation between the groups. One hundred forty ETs were performed in patients assigned to protocol A and 142 in patients assigned to protocol B. The clinical pregnancy rate in group A was 12.9% compared with 24.6% in group B. CONCLUSION(S) The administration of progesterone before oocyte retrieval is associated with a lower pregnancy rate than the administration of progesterone after oocyte retrieval.

Does ethnicity influence in vitro fertilization (IVF) birth outcomes

OBJECTIVE To determine if ethnicity influences IVF birth outcome. DESIGN Retrospective cohort study. SETTING University-based IVF program. PATIENT(S) All African American women (n = 71) and Caucasian women (n = 180) who underwent initial fresh, nondonor IVF/embryo transfer (ET) cycles between January 1, 2004 and December 31, 2005. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Gonadotropin dose, duration of stimulation, peak estradiol levels, oocyte yield, implantation, clinical pregnancy, and live birth rates. RESULT(S) African American women generated significantly fewer embryos than Caucasian women (5.3 +/- 3.7 vs. 6.6 +/- 4.8) despite having similar ages, day 3 FSH, peak estradiol levels, length of stimulation, and number of oocytes retrieved. In addition, compared with Caucasian women, African American had significantly greater body mass indices (26.5 +/- 5.2 vs. 23.7 +/- 4.8) and required significantly more total gonadotropin (IU) (4,791 +/- 2,161 vs. 3,725 +/- 2,005) for ovarian stimulation. African American women were more likely to have uterine fibroids (21% vs. 3%) and tubal factor infertility (23% vs. 9%). Caucasian women were more likely to have unexplained infertility (53% vs. 32%). Differences in embryo yield between patient groups persisted after accounting for differences in infertility diagnosis and prevalence of fibroids. Biochemical, clinical pregnancy, and live birth rates as well as implantation rates (number of sacs visualized/number of embryos transferred) did not significantly differ between groups. CONCLUSION(S) Although African Americans yield fewer embryos than Caucasian women with IVF, these ethnic groups do not seem to differ with regard to IVF pregnancy outcomes.

Ultrasound prediction of follicle volume: is the mean diameter reflective? *

OBJECTIVE To evaluate the relationship between 2 dimensional sonographic measurement of ovarian follicles and their actual volume. DESIGN Prospective clinical study. SETTING The in vitro fertilization (IVF) program of a University based, tertiary care hospital. PATIENTS AND INTERVENTIONS Sonographic categorization by shape, and measurement of 96 individual ovarian follicles immediately prior to aspiration for IVF. Each follicle was aspirated under direct ultrasound guidance and the volume recorded. The 96 follicles were visualized in a total of 14 patients from whom 2 to 27 oocytes were obtained. MAIN OUTCOME MEASURE Total volume of each follicle. RESULTS Round and polygonal follicles exhibited a highly significant relationship between sonographically measured mean diameter and total follicle volume. The volume of follicles that were categorized as ellipsoid was not predicted by measurement of the longest diameter, shortest diameter or mean diameter. CONCLUSION The mean diameter of round and polygonal follicles accurately predicts total follicular volume. However, clinical decisions in ovulation induction should be modified when the follicle shape is predominantly ellipsoid because the traditionally held belief that the sonographic measurement of the follicular diameter correlates with the follicular volume does not apply in those circumstances.

Failed fertilization after intracytoplasmic sperm injection: the extent of paternal and maternal chromatin decondensation

OBJECTIVE To determine the extent of paternal and maternal chromatin decondensation in unfertilized eggs after intracytoplasmic sperm injection (ICSI). DESIGN Eggs that failed to show two pronuclei (2-PN) 48 hours after ICSI were studied at two different time intervals: at ICSI program inception (group A) and after 8 months (group B). PATIENT(S) Forty-nine patients undergoing IVF cycles. MAIN OUTCOME MEASURE(S) The unfertilized eggs were studied by chromatin staining. RESULT(S) The average fertilization rate from all ICSI cycles in these two groups was 45%. The fertilization rates in groups A and B were 35% and 59%, respectively. In group A, 65% of the unfertilized eggs were characterized by condensed sperm chromatin with 11% showing partial decondensation. In group B, only 28% of the unfertilized eggs demonstrated condensed sperm chromatin, whereas 45% were partially decondensed. In these two groups, no sperm chromatin was detected in 24% of the unfertilized eggs. The maternal chromatin remained at metaphase II in 84% of all unfertilized eggs analyzed. CONCLUSION(S) These observations suggest that the technical problem of deposition of the sperm inside the egg is not the major cause of failure of fertilization rates in ICSI cycles. Rather, it is likely to be the failure to complete both the maternal and paternal chromatin transitions that occur with normal fertilization.

Technical and physiological aspects associated with the lower fertilization following intra cytoplasmic sperm injection (ICSI) in human

The fertilization rates with ICSI range from 30% to 70% and suggest that, despite injecting sperm into mature oocytes, significant fertilization failure still occurs in humans. The objective of this study was to determine technical and physiological factors which may contribute to lower fertilization following ICSI. Eggs that failed to show two pronuclei (PN) 48 hours after ICSI were studied at two different time intervals: at ICSI program inception (group A) and after 8 months (group B). The eggs were analyzed by staining with DNA fluorochromes, Hoescht 33258 and DAPI. The extent of sperm head as well as maternal chromatin decondensation in unfertilized ICSI eggs was determined by high resolution fluorescence microscopy. The average fertilization rate (FR) from all ICSI cycles in these two groups was 45%. The FR in Groups A and B were 35% and 59%, respectively (P < 0.05). In Group A, 65% of the unfertilized eggs were characterized by condensed sperm chromatin with 11% showing partial decondensation. In Group B, only 28% of the unfertilized eggs demonstrated condensed sperm chromatin while 45% were partially decondensed. Sperm chromatin was not detected in 24% of all unfertilized eggs studied. The maternal chromatin remained at metaphase II in 84% of all unfertilized eggs analyzed. These observations suggest that the technical problem of deposition of the sperm inside the egg is not the major cause for failure of fertilization rates in ICSI cycles. The increased percentage of eggs undergoing sperm head decondensation may be related to subtle changes in technique as experience is gained over time. The failure of sperm head decondensation in some of the ICSI eggs may be associated with cytoplasmic immaturity but not nuclear maturity.

Profound teratospermia does not influence sex chromosomal aneuploidy rate in in vitro fertilization–preimplantation genetic diagnosis cycles

Because spermatocyte meiotic error results in embryonic sex chromosomal aneuploidy, it is speculated that teratospermia correlates with increased embryonic sex chromosomal abnormalities. Our findings contradict this theory, suggesting that morphology does not correlate with sex chromosomal genotype.

The use of lead follicle diameter to initiate gonadotropin-releasing hormone antagonist does not affect in vitro fertilization clinical pregnancy, implantation, or live birth rates: a prospective, randomized study

In this prospective, randomized study, waiting for the lead follicle to reach 14 mm before initiating GnRH antagonist administration effectively prevents an LH surge and ovulation during an IVF cycle. Furthermore, delaying GnRH start until the dominant follicle reaches 14 mm neither impacts the clinical pregnancy, implantation, or live birth rates nor increases the incidence of severe ovarian hyperstimulation syndrome.

A zona biochemical change and spontaneous cortical granule loss in eggs that fail to fertilize in in vitro fertilization**Mouse studies were supported by grant HD 24191 (T.D.) and the Center for Reproductive Research (P30 HD 28897) at Tufts University, from the National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.

OBJECTIVE To determine the extent of biochemical changes in the zona pellucida (ZP) and cortical granule release in eggs failing to fertilize in IVF. DESIGN After insemination, unactivated eggs without two pronuclei (PN) were studied by high resolution microscopy and fluorescent probes to determine cortical granule density and meiotic stage. After ZP isolation, proteins from individual ZPs were biotinylated, electrophoresed, and visualized by western blots with avidin-chemiluminescence. Controls included mouse unfertilized and fertilized eggs and human germinal vesicle stage oocytes and > or = 3PN eggs. SETTING University medical center and hospital tertiary care IVF-ET program. RESULTS Many of the unfertilized eggs were in metaphase and had relatively low cortical granule densities indicative of cortical granule loss. Approximately one half of the ZPs showed evidence of biochemical hardening with a modification of a 90 to 100 x 10(3) molecular (weight) ratio (M(r)) ZP protein. The 3PN eggs had few cortical granules and their ZPs had a pronounced 90 to 100 x 10(3) M(r) modification that was not detected in germinal vesicle stage ZP controls. CONCLUSION The observed changes in ZP biochemistry and cortical granule quantitation demonstrate that failed fertilization is frequently associated with spontaneous cytoplasmic, but not nuclear, activation. In affected eggs, these changes could prevent fertilization in routine IVF and in cases of reinsemination. The relationship of changes in the ZP and cortical granules to infertility and in vitro culture requires further investigation. The 90 to 100 x 10(3) M(r) ZP protein modification can be detected biochemically with 1/10 of a human ZP.