Denny Sakkas

Yale University, New Haven, Connecticut, United States

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Publications (172)491.38 Total impact

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    ABSTRACT: Genes critical for fertility are highly conserved in mammals. Interspecies DNA sequence variation, resulting in amino-acid substitutions and post-transcriptional modifications, including alternative splicing, are a result of evolution and speciation. The mammalian Follicle Stimulating Hormone Receptor (FSHR) gene encodes distinct species-specific forms by alternative splicing. Skipping of exon 2 of the human FSHR was reported in women of North American origin and correlated with low response to ovarian stimulation with exogenous FSH. To determine whether this variant correlated with low response in women of different genetic backgrounds, we performed a blinded retrospective observational study in a Turkish cohort. Ovarian response was determined as low, intermediate or high according to retrieved oocyte numbers after classifying patients in 4 age groups (<35, 35-37, 38-40, >40). Cumulus cells collected from 96 women undergoing IVF/ICSI following controlled ovarian hyperstimulation revealed four alternatively spliced FSHR products in seven patients (8%): exon 2 deletion in four patients; exon 3 and exons 2+3 deletion in one patient each, and a retention of an intron 1 fragment in one patient. In all others (92%) splicing was intact. Alternative skipping of exons 2, 3 or 2+3 were exclusive to low responders and was independent of the use of agonist or antagonist. Interestingly, skipping of exon 3 occurs naturally in the ovaries of domestic cats - a good comparative model for human fertility. We tested the signaling potential of human and cat variants after transfection in HEK293 cells and FSH stimulation. None of the splicing variants initiated cAMP signaling despite high FSH doses, unlike full-length proteins. These data substantiate the occurrence of FSHR exon skipping in a subgroup of low responders and suggest that species-specific regulation of FSHR splicing plays diverse roles in mammalian ovarian function.
    Molecular Human Reproduction 03/2014; · 4.54 Impact Factor
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    ABSTRACT: During spermatogenesis, gene expression is regulated post-transcriptionally by modification of the poly-adenylation tail length of repressed mRNAs. In a previous study, we demonstrated that the expression of two mRNA binding proteins, embryonic poly(A)-binding protein (EPAB) and somatic cytoplasmic poly(A)-binding protein (PABPC1), is spatially and temporally controlled during male germ-cell development, suggesting that they may modulate gene expression in different spermatogenic cell types (Ozturk et al. 2012).We therefore evaluated the fertility of homozygous Epab knockout male mice (Epab-/-) to further define the importance of EPAB during spermatogenesis. Mol. Reprod. Dev. © 2014 Wiley Periodicals, Inc.
    Molecular Reproduction and Development 03/2014; · 2.81 Impact Factor
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    ABSTRACT: What is the value of embryo selection by metabolomic profiling of culture medium with near-infrared (NIR) spectroscopy as an adjunct to morphology, compared with embryo selection by morphology alone, based on an individual patient data meta-analysis (IPD MA)? The IPD MA indicates that the live birth rate after embryo selection by NIR spectroscopy and morphology is not significantly different compared with the live birth rate after embryo selection by morphology alone. Retrospective proof of principle studies has consistently shown that high NIR viability scores are correlated with a high implantation potential of embryos. However, randomized controlled trials (RCTs) have generally shown no benefit of the NIR technology over embryo morphology, although there have been some conflicting results between pregnancy outcomes on different days of embryo transfer. This IPD MA included all existing RCTs (n = 4) in which embryo selection by morphology was compared with embryo selection by morphology and the use of NIR spectroscopy of spent embryo culture medium by the Viametrics-E(™). Searches of PubMed, the Cochrane Library and the WHO International Clinical Trials Registry were conducted and the sole manufacturer of the Viametrics-E(™) was consulted to identify clinics where an RCT comparing embryo selection by morphology to embryo selection by morphology and the use of the Viametrics-E(™) (NIR viability score) was performed. A total of 20 citations were potentially eligible for inclusion, two of which met the inclusion criteria. The manufacturer of the Viametrics-E(™) provided two additional clinical sites of use. In total, four RCTs were identified as eligible for inclusion. The IPD MA was based on a fixed effect model due to the lack of heterogeneity between included studies. Differences between study groups were tested and reported using logistic regression models adjusted for significant confounders. The pooled analysis of the primary outcome led to a total sample size of 924 patients: 484 patients in the control group (embryo selection by morphology alone) and 440 patients in the treatment group (embryo selection by morphology plus NIR spectroscopy). The live birth rates in the control group and the NIR group were 34.7% (168 of 484) and 33.2% (146 of 440), respectively. The pooled odds ratio (OR) was 0.98 [95% confidence interval (CI) 0.74-1.29], indicating no difference in live birth rates between the two study groups. The data of the four studies showed no significant heterogeneity (I(2) = 26.2% P = 0.26). The multivariate regression analysis including all confounders show that maternal age (OR 0.90, 95% CI 0.87-0.94) and the number of previous IVF cycles (OR 0.83, 95% CI 0.71-0.96) were significantly related to live birth. The study group (i.e. embryo selection by morphology or embryo selection by morphology plus NIR) was not related to live birth (OR 0.97, 95% CI 0.73-1.29). The availability of at least two similar best quality embryos as an inclusion criterion prior to transfer in the two largest RCTs might have caused a selection bias towards a better prognosis patient group. There is at present no evidence that NIR spectroscopy of spent embryo culture media in its current form can be used in daily practice to improve live birth rates.
    Human Reproduction 01/2014; · 4.67 Impact Factor
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    ABSTRACT: To determine (a) the correlation between follicular sizes, oocyte maturity, normal fertilization rate, cleavage and embryo quality; and (b) to establish whether oocytes recovered with or without follicular flushing have different developmental competence. Prospective observational study. Academic medical center. Forty nine cycles (37 ICSI and 12 IVF). Measurement of 360 follicular diameters on the day of egg retrieval and classification into three groups Group A (mean diameter 12-14.5 mm.), group B (mean diameter 15-18 mm.) and group C (diameter >18.5 mm.). Correlation between follicular size at the time of retrieval and oocyte maturity, fertilization and cleavage rate in 226 oocytes (163 ICSI and 63 IVF). Developmental competence of oocytes retrieved with flushing versus non flushing. Almost all (99 %) of the oocytes recovered from follicles of group C were in metaphase II as opposed to 80 % in group A and 81 % in group B (p < 0.01). Overall there was a progressive and significant increase in fertilization rates from group A follicles to group C (47 % vs. 67 %, p 0.05). Overall 53 % of oocytes retrieved from group A follicles showed either no fertilization or abnormal fertilization versus 27 % in group C (p 0.05). The oocyte recovery rate with follicular flushing improved from group A to group B and to group C follicles (65 % vs. 49 % vs.37 % respectively p < 0.01). There were no differences in rates of immature oocyte, fertilization, abnormal or not fertilization and cleavage. The results of this study shows that: a) Follicles larger than 18 mm at retrieval have consistently mature oocytes with a higher rate of fertilization; b) Small size follicles are still capable of containing mature oocytes, but their rate of abnormal or no fertilization is high; c) Oocytes recovered with flushing are still able to produce embryos with full developmental competence.
    Journal of Assisted Reproduction and Genetics 11/2013; · 1.82 Impact Factor
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    ABSTRACT: In mammalian species, acquisition of sperm fertilization competence is dependent on the phenomenon of sperm capacitation. One of the key elements of capacitation is protein tyrosine phosphorylation (TP) in various sperm membrane regions. In previous studies performed, the pattern of TP was examined in human sperm bound to zona pellucida of oocytes. In the present comparative study, TP patterns upon sperm binding to the zona pellucida or hyaluronic acid (HA) were investigated in spermatozoa arising from the same semen samples. Tyrosine phosphorylation, visualized by immunofluorescence, was localized within the acrosomal cap, equatorial head region, neck, and the principal piece. Tyrosine phosphorylation has increased in a time-related manner as capacitation progressed, and the phosphorylation pattern was identical within the principal piece and neck, regardless of the sperm bound to the zona pellucida or HA. Thus, the data demonstrated that the patterns of sperm activation-related TP were similar regardless of the spermatozoa bound to zona pellucida or HA. Further, sperm with incomplete development, as detected by excess cytoplasmic retention, failed to exhibit TP.
    Reproductive sciences (Thousand Oaks, Calif.) 09/2013; · 2.31 Impact Factor
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    ABSTRACT: PURPOSE: The aim of this study was to analyze the outcomes of IVF/ICSI cycles in women aged 43 and beyond. METHODS: Retrospective analysis of clinical pregnancy and live birth rates in168 fresh, non donor, ART cycles performed in two Connecticut university IVF programs. RESULTS: In women of 43 and 44 years the overall clinical pregnancy and live birth rates were 8.3 % and 5.3 % per initiated cycle, respectively. There were no clinical pregnancies in women ≥45 years old. First cycle characteristics were not different from repeated cycles in terms of duration of ovulation induction, number of collected oocytes and transferred embryos (p > 0.05). CONCLUSIONS: Pregnancies can still be achieved with IVF/ICSI up to the age of 44. Since most pregnancies occurred within the first 3 cycles, another attempt may be a reasonable option before resorting to oocyte donation for patients who failed two previous cycles. Women 45 years and beyond do not benefit from ART procedures using their own oocytes.
    Journal of Assisted Reproduction and Genetics 03/2013; · 1.82 Impact Factor
  • Denny Sakkas
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    ABSTRACT: The increasing focus on developing new tools to more accurately diagnose and select individual sperm before intracytoplasmic sperm injection will allow us to counsel and treat couples with greater confidence and efficiency. Current sperm selection techniques are based on the premise that if an ejaculated spermatozoon has cleared spermatogenesis with the correct morphology and/or membrane properties then it is most likely normal. Techniques that are designed to prepare a clean "normal" sperm population or that assist in selecting an individual "normal" spermatozoon are currently being investigated. The use of techniques, including density-gradient preparation, electrophoretic separation, microfluidics, high-magnification sperm morphology selection, and hyaluronic acid binding, is discussed. The research evidence that supports the interrelated developmental and genetic integrity of the selected sperm, particularly sperm DNA damage and clinical outcome evidence are presented.
    Fertility and sterility 01/2013; · 3.97 Impact Factor
  • Kathryn C Humm, Denny Sakkas
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    ABSTRACT: The well documented increase in age that women conceive their first child has detracted from a similar change observed in males. As both males and females decide to conceive later, the question of whether this may impact their fertility individually and as a couple becomes even more crucial. A paternal age of over 40 years at the time of conception is a frequently quoted male age threshold, however, currently there is no clearly accepted definition of advanced paternal age or even a consensus on the implications of advancing male age. In this paper, we review some of the potential risks to the offspring of advancing male age and examine. The data available regarding pregnancy outcomes based on paternal age in both the fertile and infertile populations. Within the infertile population specifically, we examine the association between male age and outcomes based on treatment modality, including intrauterine insemination (IUI), in vitro fertilization (IVF), and donor oocyte IVF. Finally, we discuss the various mechanisms by which male age may impact sperm and fertility potential, including sperm DNA damage.
    Fertility and sterility 01/2013; 99(1):30-6. · 3.97 Impact Factor
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    ABSTRACT: Metabolomics was introduced in human in vitro fertilization (IVF) for noninvasive identification of viable embryos with the highest developmental competence. To determine whether embryo selection using a commercial version of metabolomic analysis leads to increased implantation rates (IRs) with fetal cardiac activity (FCA) compared with morphology evaluation alone. Randomized controlled trial from April to December 2010 at a private IVF unit. The study was terminated prematurely due to the market withdrawal of the instrument. IVF patients ≥18 and ≤43 years with ≥4 × 2PN were randomly allocated to metabolomic analysis combined with embryo morphology (ViaMetrics-E; metabolomics + morphology group) or embryo morphology alone (morphology group). Cycles with frozen embryos, oocyte donations, or testicular biopsy were excluded. Categorical and continuous data were analyzed for statistical significance using 2-tailed Fisher's exact test and t-test, respectively. Statistical significance was accepted when P > 0.05. A total of 125 patients were included in the study; 39 patients were allocated to metabolomics + morphology group and 86 patients to morphology group. Patients were stratified according to the day of embryo transfer (Days 2, 3, or 5). IRs with FCA were similar for Days 2 and 3 transfers in both groups. For Day 5 transfers, IRs with FCA were significantly higher in the metabolomics + morphology group (46.8% vs. 28.9%; P = 0.041; 95% confidence intervalp [CI]: 1.09-34.18). Pregnancy and live births rates were similar for Days 2, 3, and 5 in both groups. The study was terminated early following the voluntary market withdrawal of ViaMetrics-E in December 2010. Metabolomic analysis using the commercial near-infrared (NIR) instrument does not appear to have a beneficial effect on pregnancy and live births, with improvement in IR with FCA for Day 5 transfers. However, no solid conclusions can be reached due to the lack of adequate study power.ClinicalTrials.gov Identifier: NCT01490515.
    Journal of Human Reproductive Sciences 01/2013; 6(2):133-139.
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    ABSTRACT: The objective of this retrospective analysis was to assess whether the outcomes of fresh blastocyst transfer cycles are predictive of the chances for pregnancy and live birth in subsequent frozen blastocyst transfer cycles using sibling embryos from the same retrieval. Clinical pregnancy rate (CPR) and live birth rate (LBR) per fresh and frozen blastocyst transfer were assessed. All subgroups had similar patient and cycle characteristics. Overall, CPR and LBR in fresh cycles were 44% and 29%, and in frozen were 34% and 30%, respectively. However, the CPR and LBR in frozen cycles were significantly higher in patients who were not pregnant with their fresh cycles (CPR 43% versus 22%, P = 0.01; and LBR 36% versus 17%, P = 0.03, respectively). When fresh cycles are unsuccessful, the remaining frozen blastocysts of the same cohort have the same chance of success in producing a clinical pregnancy as the fresh cycle (43% versus 44%). Frozen cycles following successful fresh cycles have significantly lower CPR and LBR. These data reinforce the concept that only a few embryos per cohort are competent for a live birth. During IVF cycles, many patients are fortunate enough to have excess high-quality embryos remaining after their embryo transfer. These embryos can be frozen, or cryopreserved, for later transfer. The transfer of cryopreserved embryos increases the cumulative success rates after a single IVF stimulation. Many studies have examined success rates such as clinical pregnancy rate and live birth rate in frozen embryo transfer cycles. While these frozen embryo transfer cycles have excellent success rates, they are significantly lower than success rates in cycles where a “fresh”, non-frozen, embryo is transferred. Few studies have carefully examined the impact of the result of the fresh embryo transfer (whether the patient became pregnant or not) on subsequent frozen embryo transfer success. Here we show that women who are not pregnant after a fresh embryo transfer have higher success rates in subsequent frozen embryo transfer cycles that use frozen embryos generated during a single IVF cycle. In these women (not pregnant after a fresh embryo transfer), frozen embryo transfer pregnancy rates are the same as rates using fresh embryos.
    Reproductive biomedicine online 01/2013; · 2.68 Impact Factor
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    ABSTRACT: STUDY QUESTION: Is the selection of a single Day 3 embryo by metabolomic profiling of culture medium with near-infrared (NIR) spectroscopy as an adjunct to morphology able to improve live birth rates in IVF, compared with embryo selection by morphology alone? SUMMARY ANSWER: The live birth rate after embryo selection by NIR spectroscopy and morphology is not significantly different compared with the live birth rate after embryos were selected by morphology alone. WHAT IS KNOWN ALREADY: The elevated incidence of pregnancy and neonatal problems associated with a high-twinning rate after IVF can only be successfully reduced by the transfer of one embryo. Current embryo assessment methods are unable to accurately predict the reproductive potential of an individual embryo. Today, a number of techniques are said to be more accurate at selecting the best embryo. One of these new technologies is metabolomic profiling of spent embryo culture media with the use of NIR spectroscopy. STUDY DESIGN, SIZE AND DURATION: A double-blind, randomized controlled trial was conducted between 2009 and 2011, and included 417 couples undergoing IVF with a single embryo transfer. Randomization was performed centrally just before Ovum Pick-Up (OPU), using a computerized randomization program. Both patient and physician were unaware of the treatment allocation. To ensure blinding, the allocations were placed in consecutively numbered, opaque envelopes. Patients were randomized (1:1) into either the control group (embryo selection by morphology only) or the treatment group (embryo selection by morphology plus NIR spectroscopy of embryo culture medium). PARTICIPANTS/MATERIALS, SETTING AND METHODS: At OPU, 208 patients were randomized to the morphology only group and 209 patients were randomized to the morphology plus viability score group. On Day 3, 163 patients in the control group and 146 patients in the treatment group met the inclusion criteria. The study was conducted in an academic hospital with IVF laboratory and three non-academic hospitals. MAIN RESULTS AND THE ROLE OF CHANCE: Patient demographics and baseline characteristics were distributed equally over the two groups, except for embryo fragmentation, which was significantly higher in the treatment group. In the intention to treat analysis, the live birth rates were 31.7 and 26.8% for the control group and the treatment group, respectively (relative risk 0.84; 95% confidence interval 0.63-1.14, P=0.27). In the per protocol analysis, the live birth rates were 31.3 and 29.5% for the control group and the treatment group, respectively (relative risk 0.94; 95% confidence interval 0.67-1.32, P=0.73). For the treatment group, the embryological technician's independent choice (by morphology) of which embryo to transfer was recorded 138 times. In 75.4% (104 of 138) of the transfers, the embryo with the best morphology did not have the highest viability score. The live birth rate of these 104 transferred embryos was 30.8%. LIMITATIONS, REASONS FOR CAUTION: A possible limitation of our study is the pre-selection of all embryos by morphology and dividing the cohort of available embryos into two groups: good quality embryos and poor quality embryos. As a consequence, we have probably selected for a better prognosis patient group. WIDER IMPLICATIONS OF THE FINDINGS: To avoid the use of incompetent embryo selection tools at the expense of the patient, an evidence-based proof of clinical usefulness is essential before the implementation of new diagnostic tools in IVF laboratories. TRIAL REGISTRATION NUMBERS: Dutch Trial Registry, registry number NTR1178.
    Human Reproduction 05/2012; 27(8):2304-11. · 4.67 Impact Factor
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    ABSTRACT: Gene expression during oocyte maturation and early embryogenesis up to zygotic genome activation requires translational activation of maternally-derived mRNAs. EPAB [embryonic poly(A)-binding protein] is the predominant poly(A)-binding protein during this period in Xenopus, mouse and human. In Xenopus oocytes, ePAB stabilizes maternal mRNAs and promotes their translation. To assess the role of EPAB in mammalian reproduction, we generated Epab-knockout mice. Although Epab(-/-) males and Epab(+/-) of both sexes were fertile, Epab(-/-) female mice were infertile, and could not generate embryos or mature oocytes in vivo or in vitro. Epab(-/-) oocytes failed to achieve translational activation of maternally-stored mRNAs upon stimulation of oocyte maturation, including Ccnb1 (cyclin B1) and Dazl (deleted in azoospermia-like) mRNAs. Microinjection of Epab mRNA into Epab(-/-) germinal vesicle stage oocytes did not rescue maturation, suggesting that EPAB is also required for earlier stages of oogenesis. In addition, late antral follicles in the ovaries of Epab(-/-) mice exhibited impaired cumulus expansion, and a 8-fold decrease in ovulation, associated with a significant down-regulation of mRNAs encoding the EGF (epidermal growth factor)-like growth factors Areg (amphiregulin), Ereg (epiregulin) and Btc (betacellulin), and their downstream regulators, Ptgs2 (prostaglandin synthase 2), Has2 (hyaluronan synthase 2) and Tnfaip6 (tumour necrosis factor α-induced protein 6). The findings from the present study indicate that EPAB is necessary for oogenesis, folliculogenesis and female fertility in mice.
    Biochemical Journal 05/2012; 446(1):47-58. · 4.65 Impact Factor
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    ABSTRACT: This study investigated if metabolomic profiling of culture media using near infrared (NIR) spectroscopy was related to live-birth rates after single-embryo transfer of frozen-thawed embryos. Analysis of culture media of frozen-thawed embryos was performed by NIR spectroscopy. A viability score was calculated using a predictive multivariate algorithm of fresh day-5 embryos with known pregnancy outcomes. This algorithm generated with fresh day-5 embryos could help to identify the live-birth group from the no live-birth group. Multivariable regression models that tested the predictive ability of the viability score for live birth showed an odds ratio in the crude analysis of 1.50 (P=0.008), after adjustment for embryo morphology, 1.44 (P=0.022), and after adjustment for all variables, 1.71 (P=0.005); based on a 0.1 step increase in viability scores. In conclusion, higher viability scores resulted in higher live-birth rates. An algorithm generated from fresh embryos might be used to predict viability of frozen-thawed embryos. Frozen-thawed embryos have different metabolic activity which is related to implantation potential. Therefore, this method might be useful to select the best embryo for transfer within a group of embryos with similar morphology.
    Reproductive biomedicine online 12/2011; 23(6):769-76. · 2.68 Impact Factor
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    ABSTRACT: Near infrared (NIR) spectroscopy is a technology proposed to facilitate non-invasive screening for the most optimal human embryo for uterine transfer. It has been proposed that the NIR spectral profile of an embryo's spent culture medium can be used to generate a viability score that correlates to implantation potential. As the initial proof of principle studies were all retrospective, our aim was to investigate whether NIR spectroscopy on spent embryo culture medium in an on-site, prospective setting could improve the ongoing single embryo transfer (SET) pregnancy rate after Day 2 and 5 transfers. We conducted a single-centre, double-blinded, randomized controlled trial in which the NIR group was compared with a control group. The primary outcome was the clinical pregnancy rate after 6-7 weeks of gestation per randomized patient. In the control group embryo selection was based only on traditional morphological evaluation while in the treatment group NIR spectroscopy was added to the morphological evaluation. The study was terminated early as the analysis of the Data Safety Monitoring Board showed a very low conditional power of superiority for the primary outcome. Of the 752 patients calculated to be included in the study, 164 and 163 patients were randomized into the NIR and control groups, respectively. No significant difference in the ongoing pregnancy rate per randomized patient was found between the NIR and the control group, 34.8 versus 35.6%, (P= 0.97). The proportional difference between the study groups mean was -0.8% (95% confidence interval -11.4 to 10.2). This study shows that adding NIR spectroscopy, in its present form, to embryo morphology does not improve the chance of a viable pregnancy when performing SET. The NIR technology appears to need further development before it can be used as an objective marker of embryo viability. CLINICAL TRIALS IDENTIFIER: ISRCTN23817363.
    Human Reproduction 11/2011; 27(1):89-96. · 4.67 Impact Factor
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    ABSTRACT: To compare the impact of mandated IVF insurance coverage on ET practices and resulting multiple pregnancy rates. Retrospective analysis of all fresh, nondonor IVF cycles performed in the United States in 2006. United States. A total of 91,753 fresh, nondonor IVF cycles in the United States. None. Pregnancy rate, live-birth rate, embryos transferred, multiple pregnancy rate. Overall, nonmandated states had a significantly higher pregnancy rate (38.8% vs. 35%) and live-birth rate (32.2% vs. 29.1%) than mandated states. Nonmandated states also had a significantly higher twin rate (28.1% vs. 26%) and triplet rate (3.9% vs. 3.4%). The average number of embryos transferred was also significantly higher in nonmandated states (2.6 vs. 2.2). These findings were more pronounced in the <35 and 35-37 age groups. In the last 8 years, despite a reduction in the average number of embryos transferred and multiple pregnancy rates, there is a continued association between mandated IVF coverage, the transfer of fewer embryos, and lower rates of multiple pregnancies and births, particularly in the younger age groups.
    Fertility and sterility 03/2011; 95(3):964-9. · 3.97 Impact Factor
  • Hasan M. El-Fakahany, Denny Sakkas
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    ABSTRACT: The term apoptosis refers to a morphologically distinct form of cell death that plays a major role during the normal development and homeostasis of multicellular organisms. This mode of cell death is a tightly regulated series of energy-dependent molecular and biochemical events orchestrated by a genetic program. Apoptosis is either developmentally regulated (launched in response to specific stimuli, such as deprivation of survival factors, exposure to ionizing radiation and chemotherapeutic drugs, or activation by various death factors and their ligands) or induced in response to cell injury or stress. Apoptosis is also being recognized in the pathogenesis of many diverse human diseases including cancer, acquired immune deficiency syndrome, neurodegenerative disorders, atherosclerosis, and cardiomyopathy. Maintaining the homeostatic relationship between apoptosis and cell proliferation is important for tissue development and degeneration. Decreased apoptosis may lead to neoplasia, whereas increased apoptosis may lead to a dystrophic condition.
    01/2011;
  • Fertility and Sterility - FERT STERIL. 01/2011; 96(3).
  • Fertility and Sterility - FERT STERIL. 01/2011; 96(3).
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    ABSTRACT: Introduction: Metabolomics was recently introduced in human IVF for non-invasive identification of viable embryos with the highest developmental competence. The aim of the present preliminary study was to determine whether embryo selection using non-invasive metabolomic profiling as an adjunct to morphology leads to increased pregnancy and implantation rates compared to routine morphological embryo evaluation alone.Materials and Methods: A prospective study was conducted from April to December 2010, at Eugonia Unit of Assisted Reproduction in Athens on 125 IVF patients with at least 4 fertilized oocytes (2PN). Patients underwent ovarian stimulation using GnRH agonist or antagonist protocols and oocyte retrieval was performed 36 hours after administration of hCG. Oocytes were fertilized by IVF or ICSI and fertilization was assessed 16–20 hours post insemination. Embryos were individually cultured in 25 μl drops of sequential culture media (ISM1/BlastAssist;Origio,Denmark). Embryo transfer was performed 2, 3, or 5 days after oocyte retrieval, depending on patient history and embryo characteristics.In the metabolomics group, 3–8 embryos of good morphology from each patient’s cohort were metabolically evaluated using Viametrics-E (Molecular Biometrics, USA) and ranked on the day of embryo transfer. 10μl of media samples of each corresponding embryo were loaded onto specialized spectrometer-compatible sample cells and measured using a near-infrared spectrometer. Analysis of each sample and corresponding control media was performed to provide the Viability Score. The Viability Score represents an assessment of specific biomarkers, corresponding to unique functional groups of molecules, and is generated using specific algorithms for Day 2, 3 and 5 transfers. The resulting viability score is a quantitative measurement of an embryo’s potential to establish a pregnancy with fetal cardiac activity (FCA).Patients were allocated (ratio ViaMetrics-E:routine morphology = 1:2) to embryo selection based either on the viability score combined with embryo morphology (n = 39, metabolomics group) or embryo morphology only (n = 86, control group). One to four embryos with the highest viability score were selected for transfer, depending on embryo quality and patient age. Categorical and continuous data were analyzed for statistical significance using 2-tailed Fisher’s exact test and unpaired t -test respectively.Results: Mean (± SD) patient age (years) (34.5 ± 4.7 vs 35.7 ± 4.4), BMI (kg/m2) (25.1 ± 5.1 vs 25.3 ± 4.2), years of infertility (3.5 ± 2.1 vs 4.0 ± 3.6), number of previous IVF attempts (1.2 ± 1.6 vs 1.5 ± 1.9), basal FSH (mIU/ml) (7.6 ± 2.5 vs 8.2 ± 2.7), number of oocytes retrieved (17.2 ± 7.3 vs 15.1 ± 6.0), number of fertilized oocytes (9.9 ± 4.0 vs 8.4 ± 4.1) and number of embryos transferred (2.6 ± 0.7 vs 2.9 ± 0.6) per patient were similar in the Viametrics-E metabolomics and control group respectively. Biochemical pregnancy rates [28/39 (71.8%) vs 57/86 (66.3%)], and clinical pregnancy rates [21/39 (53.9%) vs 41/86 (47.7%)] were improved between the metabolomics and control groups but did not differ significantly. A total of 359 embryos were transferred (metabolomics: n = 102 embryos; control: n = 257 embryos). Implantation rates (37.3% vs 25.7%; p = 0.04) and implantations with FCA (32.4% vs 21.4%; p = 0.04) were significantly higher in the metabolomics group compared to controls.In the metabolomics group, a total of 196 embryos were analyzed and 102 embryos were transferred. The mean viability score of embryos selected for transfer (0.6 ± 0.2) was significantly higher (p < 0.0001) compared to embryos not selected for transfer (0.3 ± 0.2).Conclusion: The present preliminary trial indicates increased implantation rates with positive FCA following embryo selection based on non-invasive metabolomic analysis. The method may provide an objective secondary level of assessment as an adjunct to embryo morphology by ranking the embryos of a patient's cohort. However, following the voluntary market withdrawal/recall of ViaMetrics-E in December 2010, the results presented here should be viewed with caution. At the time of writing, an upgraded version is awaited, which will require further stringent assessment and validation, in a single embryo transfer setting.
    Human Reproduction. 01/2011; 26(Suppl. 1):i86.

Publication Stats

5k Citations
491.38 Total Impact Points

Institutions

  • 2001–2014
    • Yale University
      • Department of Obstetrics, Gynecology and Reproductive Sciences
      New Haven, Connecticut, United States
  • 2013
    • Beth Israel Deaconess Medical Center
      Boston, Massachusetts, United States
    • University of Massachusetts Boston
      Boston, Massachusetts, United States
    • Akdeniz University
      • Department of Histology and Embryology
      Antalya, Antalya, Turkey
  • 2001–2013
    • Yale-New Haven Hospital
      New Haven, Connecticut, United States
  • 2008
    • Reproductive Medicine Associates of New Jersey
      Somerset, New Jersey, United States
  • 2005
    • ENEA
      Roma, Latium, Italy
  • 1993–2005
    • University of Geneva
      • Department of Obstetrics and Gynaecology
      Genève, GE, Switzerland
  • 2000
    • Università degli Studi di Modena e Reggio Emilia
      • Department of Life Sciences
      Modène, Emilia-Romagna, Italy
  • 1999–2000
    • University of Birmingham
      • School of Biosciences
      Birmingham, England, United Kingdom
  • 1992
    • Monash University (Australia)
      Melbourne, Victoria, Australia