James A Grifo, M.D., Ph.D.

Is intracytoplasmic sperm injection overused?
Hodes-Wertz, Brooke; Mullin, Christine M; Adler, Alexis; Noyes, Nicole; Grifo, James A; Berkeley, Alan S
2012 Feb;187(2):602-606, Journal of urology
PURPOSE: We determined whether the use of intracytoplasmic sperm injection in couples who previously underwent intracytoplasmic sperm injection cycles elsewhere could be decreased without compromising the pregnancy rate. MATERIALS AND METHODS: At our university in vitro fertilization-embryo transfer center we retrospectively analyzed the records of 149 fresh, in vitro fertilization-embryo transfer cycles in patients who underwent intracytoplasmic sperm injection elsewhere and subsequent fertilization by insemination only (all insemination group) or half insemination and half intracytoplasmic sperm injection at our center. We compared fertilization, implantation, clinical pregnancy and live birth rates. RESULTS: The fertilization rate was 74% and 73% for the all insemination and the half intracytoplasmic sperm injection groups, respectively. In the latter group 69% of inseminated and 78% of intracytoplasmic sperm injected oocytes were fertilized. No cycle showed complete fertilization failure. No statistically significant difference in the live birth rate was found between the 2 groups. CONCLUSIONS: More stringent criteria for intracytoplasmic sperm injection do not compromise the clinical outcome and reasonable fertilization can be achieved whether or not intracytoplasmic sperm injection is performed. Thus, although intracytoplasmic sperm injection is one of the greatest advances in our field, it is overused and should only be done for clinically proven indications
— id: 149780, year: 2012, vol: 187, page: 602, stat: Journal Article,

Blastocyst culture selects for euploid embryos: Comparison of blastomere biopsy (EB) and trophectoderm biopsy (TE) for aneuploidy rates using array comparative genomic hybridization (a-CGH)
Adler A.; Lee H.-L.; Ampeloquio E.; Clarke-Williams M.; Grifo J.
2011 ;96(3 SUPPL 1):S108-S109, Fertility & sterility
OBJECTIVE: To compare the euploidy rates using whole genome amplification and a-CGH following EB and TE biopsy. DESIGN: Case controlled prospective study comparing aneuploidy, implantation and pregnancy rates following EB and TE biopsy for 24 chromosome assessment. MATERIALS AND METHODS: EB was performed on embryos with at least 4 cells and euploid embryos were either transferred or vitrified on day 5. TE biopsy was performed after a channel was made in the zona pellucida (ZP) on day 3 with a Research Instrument laser. On day 5 and 6 blastocysts that had expanded with differentiation were biopsied by removing herniated TE with the laser. Blastocysts were vitrified using Sage Vitrification Media and Biotech Cryolocks., A programmed thaw cycle was initiated using estrogen and progesterone the desired number (1-2) euploid of blasts were warmed and transferred. RESULTS: 94 patients had 870 embryos biopsied on day 3 biopsy. 6 patients had 105 blastocysts for TE biopsy. Biopsy results revealed 218/870 (25%) euploid embryos following day 3 biopsy versus 46/105 (44%) for TE biopsy (P=0.0001 Fisher exact test). 37/94 (39%) patients had a positive hCG following day 3 biopsy compared to 8/12 (66%) frozen embryo transfers after TE biopsy (P=0.0918). The implantation rate for day 3 biopsy was 47/ 115 (40%), compared to 8/19 (42%) for TE (NS). CONCLUSION: Day 3 EB takes place prior to or at the time of embryonic gene expression. Some aneuploidies do not allow embryos to progress beyond the cleavage stage. This may account for the higher euploidy rates of day 5/6 embryos. Studies have shown a greater incidence of mosaicism on day 3 that possibly corrects by the blastocyst stage. This can also account for higher euploidy rates of blastocysts and potentially a higher pregnancy and implantation rate. By screening out aneuploid embryos blastocyst culture may result in better embryo selection and better outcome compared with D3 embryos
— id: 150886, year: 2011, vol: 96, page: S108, stat: Journal Article,

Retrospective analysis of outcomes following transfer of previously cryopreserved oocytes, pronuclear zygotes and supernumerary blastocysts
Hodes-Wertz, Brooke; Noyes, Nicole; Mullin, Christine; McCaffrey, Caroline; Grifo, Jamie A
2011 Jul;23(1):118-123, Reproductive biomedicine online
Oocyte cryopreservation still bears the experimental label. Remarkable innovation in this field has led to immense improvement in clinical outcomes and has even resulted in outcomes comparable to those achieved following fresh embryo transfers. Such success has prompted this centre to investigate outcomes of cryopreservation options (oocyte versus pronuclear zygote versus supernumerary day-5 blastocyst after fresh embryo transfer). This study retrospectively analysed 200 cryopreservation cycles which were divided into three groups according to cryopreservation option, which were all cultured to blastocyst-stage post thaw/warming from January 2005 to December 2008, and compared them with 400 fresh embryo transfer cycles from the same time period. When compared with fresh embryo transfer, frozen embryo transfers originating from previously cryopreserved oocytes or pronuclear zygotes resulted in similar implantation, pregnancy and live-birth rates; however, frozen embryo transfers originating from supernumerary day-5 blastocysts resulted in lower outcomes. Thus, oocyte and/or pronuclear zygote cryopreservation appear to be the most viable options for women desiring fertility preservation. Cryopreservation of supernumerary blastocysts may lead to a slightly lower live-birth rate since the best-quality blastocysts are generally transferred during the fresh embryo transfer attempt. Despite substantial advancements in oocyte freeze-thaw methods, allowing for much improved clinical outcomes, including live birth rates comparable to those achieved following fresh embryo transfer cycles, oocyte cryopreservation still bears the experimental label. Such recent reported success with cryopreservation has prompted us to investigate the clinical outcomes of our institution's currently available cryopreservation options (oocyte versus pronuclear zygote versus supernumerary day-5 blastocyst remaining after a fresh transfer). A total of 200 cryopreservation cycles were reviewed and subdivided according to cryopreservation optioninto three groups (oocyte versus pronuclear zygote versus day-5 blastocyst cryopreservation that were remaining after a fresh transfer), and compared with 400 fresh embryo transfer cycles from January 2005 to December 2008. When compared with fresh embryo transfer cycles, frozen embryo transfers originating from previously cryopreserved oocytes or pronuclear zygotes resulted in similar implantation, pregnancy and live birth rates. However, frozen embryo transfers originating from supernumerary day-5 blastocysts resulted in lower implantation and pregnancy rates when compared with controls. Thus, oocyte and/or pronuclear zygote cryopreservation appear to be the most viable options for women desiring fertility preservation as the outcomes from these treatments were comparable to those of fresh embryo transfer treatments. In addition, cryopreservation of supernumerary day-5 blastocysts may lead to a slightly lower live birth rate since the best-quality blastocysts are generally transferred during the fresh embryo transfer attempt
— id: 135539, year: 2011, vol: 23, page: 118, stat: Journal Article,

Treatment outcomes and quality-of-life assessment in a university-based fertility preservation program: Results of a registry of female cancer patients at 2 years
Reh, Andrea E; Lu, Lucy; Weinerman, Rachel; Grifo, James; Krey, Lewis; Noyes, Nicole
2011 Jul;28(7):635-641, Journal of assisted reproduction & genetics
PURPOSE: To explore patient goals and quality of life (QOL) via a prospective registry and compare fertility preservation (FP) outcomes before, during, and after cancer therapy. METHODS: Of 35 patients entering the registry from 3/2008 to 3/2010, 29/35 completed the study survey and agreed to follow-up, and 31/35 completed treatment. Survey results and FP outcomes were analyzed. RESULTS: Most patients rated the impact of cancer treatment on fertility of highest importance at baseline and 1-year follow-up. QOL scores were overall positive at both intervals. Patients naive to any cancer treatment (n = 12) had more gametes frozen than patients with prior cancer treatment (n = 19) with no difference in age or gonadotropin dosage. For patients awaiting cancer treatment, the median time from consultation to oocyte retrieval was 25 days. Cancer treatment sequalae posed challenges to optimal FP outcomes. CONCLUSIONS: Fertility preservation remains a significant issue for cancer patients. With early reproductive endocrinologist referral, cancer treatment delay is minimized and FP outcomes are optimized
— id: 136992, year: 2011, vol: 28, page: 635, stat: Journal Article,

Effect of autoimmune thyroid disease in older euthyroid infertile woman during the first 35 days of an IVF cycle
Reh, Andrea; Chaudhry, Sonal; Mendelsohn, Felicia; Im, Shelly; Rolnitzky, Linda; Amarosa, Alana; Levitz, Mortimer; Srinivasa, Suman; Krey, Lewis; Berkeley, Alan S; Grifo, James A; Danoff, Ann
2011 Mar 1;95(3):1178-1181, Fertility & sterility
In this case-control study of euthyroid first-cycle IVF patients >/= 38 years old with singleton baby, miscarriage, biochemical pregnancy, and no pregnancy outcomes from 2005-2008, we assayed frozen serum for autoimmune thyroid disease (AITD) and thyroid function at cycle start, trigger, and 4 and 5 weeks' gestation. AITD prevalence in older infertile women was similar across clinical outcomes, and although AITD was associated with a higher baseline TSH, TSH remained within acceptable ranges, suggesting that T(4) supplementation may not affect maternal outcomes in older euthyroid AITD patients through 5 weeks gestation
— id: 138179, year: 2011, vol: 95, page: 1178, stat: Journal Article,

Derivation of Novel Genetically Diverse Human Embryonic Stem Cell Lines
Stefanova VT; Grifo JA; Hansis C
2011 Dec 28;:?-? #, Stem cells & development
Human embryonic stem cells (hESCs) have the potential to revolutionize many biomedical fields ranging from basic research to disease modeling, regenerative medicine, drug discovery and toxicity testing. A multitude of hESC lines have been derived worldwide since the first five lines by Thomson and colleagues 13 years ago, but many of these are poorly characterized, unavailable or do not represent desired traits, thus making them unsuitable for application purposes. In order to provide the scientific community with better options, we have derived twelve new hESC lines at New York University from discard genetically normal and abnormal embryos using the latest techniques. We examined the genetic status of the NYUES lines in detail as well as their molecular and cellular features and DNA fingerprinting profile. Furthermore, we differentiated our hESCs into the tissues most affected by a specific condition or into clinically desired cell types. To our knowledge, a number of characteristics of our hESCs have not previously been reported, e.g. mutation for alpha thalassemia X-linked mental retardation syndrome, linkage to conditions with a genetic component such as asthma or poor sperm morphology and novel combinations of ethnic backgrounds. Importantly, all of our undifferentiated euploid female lines tested to date did not show X chromosome inactivation, believed to result in superior potency. We continue to derive new hESC lines and add them to the NIH registry and other registries. This should facilitate the use of our hESCs and lead to advancements for patient-benefitting applications
— id: 149781, year: 2011, vol: , page: ?, stat: Journal Article,

DOES NEWLY AVAILABLE 24-CHROMOSOME (24C) PREIMPLANTATION GENETIC SCREENING (PGS) IMPROVE IVF OUTCOMES IN PATIENTS AT RISK FOR ANEUPLOIDY? FIRST YEAR'S EXPERIENCE AT A LARGE, UNIVERSITY-BASED CENTER
Devine, K.; Knopman, J.; Adler, A.; Berkeley, A.; Grifo, J.
2010 SEP ;94(4):S123-S123, Fertility & sterility
— id: 113769, year: 2010, vol: 94, page: S123, stat: Journal Article,

Delivery rate using cryopreserved oocytes is comparable to conventional in vitro fertilization using fresh oocytes: potential fertility preservation for female cancer patients
Grifo, James A; Noyes, Nicole
2010 Feb;93(2):391-396, Fertility & sterility
OBJECTIVE: To explore the use of oocyte cryopreservation as a fertility-conserving option. Cancer treatments administered during the reproductive and adolescent years can result in sterility. Previous fertility preservation efforts focused on embryo rather than oocyte storage because the latter was deemed inefficient. Recently, several large reports of healthy births resulting from the transfer of embryos derived from frozen/thawed oocytes have been published. We sought to establish an oocyte cryopreservation program at our center. DESIGN: Twenty-three oocyte cryopreservation cycles were performed. Collected oocytes were cryopreserved by either the slow or the vitrification method. Approximately 1-4 months later, a programmed cycle of thawing/warming, fertilization with intracytoplasmic sperm injection, and ET was performed; cycle and pregnancy outcomes were assessed. SETTING: University-based fertility center. PATIENT(S): Twenty-two infertile women. INTERVENTION(S): Oocyte cryopreservation. MAIN OUTCOME MEASURE(S): Oocyte survival, embryo development, pregnancy outcomes. RESULT(S): Oocyte survival, 2-pronuclei fertilization, and blastocyst formation rates were 92%, 79%, and 43%, respectively. Fourteen women became pregnant; one miscarried; 10 have delivered 13 viable infants, and three pregnancies are ongoing for an ongoing/delivered pregnancy rate of 57%. This result was not statistically different from cycles performed consecutively in age-matched controls using fresh, nonfrozen autologous or donor oocytes during a similar time period. CONCLUSION(S): Oocyte cryopreservation appears to be a viable option for fertility preservation in some centers
— id: 99203, year: 2010, vol: 93, page: 391, stat: Journal Article,

VALIDATION AND FIRST CLINICAL APPLICATION OF KARYOMAPPING FOR PREIMPLANTATION DIAGNOSIS (PGD) OF GAUCHER DISEASE COMBINED WITH 24 CHROMOSOME SCREENING
Handyside, A. H.; Grifo, J.; Prates, R.; Tormasi, S.; Fischer, J.; Munne, S.
2010 SEP ;94(4):S79-S80, Fertility & sterility
— id: 113767, year: 2010, vol: 94, page: S79, stat: Journal Article,

GENERATION AND CHARACTERIZATION OF DISEASE-SPECIFIC HUMAN EMBRYONIC STEM CELLS FROM GENETICALLY ABNORMAL EMBRYOS
Hansis, C.; Rice, C. E.; Lehmann, R.; Grifo, J. A.
2010 SEP ;94(4):S30-S30, Fertility & sterility
— id: 113763, year: 2010, vol: 94, page: S30, stat: Journal Article,

Cancer care on a continuum: Maintaining fertility after diagnosis and treatment
Knopman J.M.; Grifo J.A.; Labella P.A.; Noyes N.
2010 ;28(15 SUPPL 1):?-?, Journal of clinical oncology
Background: Early detection programs combined with improved treatment protocols have allowed cancer patients (CP) to live substantially longer lives. Therefore, quality-of-life issues, such as fertility preservation (FP), have become increasingly important. Previously, time constraints and poor success limited the effectiveness of such procedures. However, recent advancements in oocyte cryopreservation (OC) technology have made this a viable option. OC not only eliminates the need for donor gametes but also the ethical, personal and religious constraints associated with embryo freezing. The novelty of OC has limited its universality; only recently have oncologists begun to refer CP for OC with even fewer yet returning to use these oocytes. To ensure feasibility, we compared OC outcomes of CP to all women without cancer who have completed oocyte thaw (n=32). Methods: CP referred for FP underwent extensive counseling in compliance with the ASRM (2009); those electing OC were included. Ovarian stimulation was achieved with injectable gonadotropins. OC was performed using slow cooling and vitrification methods. Results: 50 CP completed an OC cycle (baseline ovarian reserve testing was normal for all CP); 6 had 1 child already. Malignant diagnoses included 22 gyn, 12 breast, 8 hematologic, 2 GI, 2 CNS and 4 other; 7/9 pts >= age 38 had breast cancer. FP treatment was completed on average in 12 +/-0.3 days. Outcome data are shown in the Table. Conclusions: OC is a novel and potentially successful FP option offering desired reproductive choice. With a dedicated team, OC can be performed expeditiously and successfully, minimizing interference with cancer treatment. As future parenthood greatly impacts quality-of-life, OC should be an integral component of FP counseling in young cancer patients. (Table presented)
— id: 112440, year: 2010, vol: 28, page: ?, stat: Journal Article,

WHAT'S THE SCORE?: A QUANTITATIVE MEANS TO ASSESS EMBRYO QUALITY (EQ)
Knopman, J. M.; Krey, L. C.; McCaffrey, C.; Noyes, N.; Hodes-Wertz, B.; Grifo, J. A.
2010 SEP ;94(4):S25-S25, Fertility & sterility
— id: 113762, year: 2010, vol: 94, page: S25, stat: Journal Article,

OOCYTE CRYOPRESERVATION: AN ALTERNATIVE MODEL FOR GAMETE DONATION
Knopman, J. M.; Noyes, N.; LaBella, P.; Licciardi, F.; Grifo, J. A.
2010 SEP ;94(4):S116-S117, Fertility & sterility
— id: 113768, year: 2010, vol: 94, page: S116, stat: Journal Article,

Cryopreserved oocytes can serve as the treatment for secondary infertility: a novel model for egg donation
Knopman, Jaime M; Noyes, Nicole; Grifo, James A
2010 May 1;93(7):2413.e7-2413.e9, Fertility & sterility
OBJECTIVE: To report the use of previously cryopreserved oocytes for the treatment of secondary infertility. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A 41-year-old woman with 18 months of secondary infertility and a previous history (age 38) of elective oocyte cryopreservation. INTERVENTION(S): Previously cryopreserved oocytes. MAIN OUTCOME MEASURE(S): Fertilization, embryo development, pregnancy, and outcome. RESULT(S): The patient achieved pregnancy and delivery following thaw of oocytes electively cryopreserved 39 months before use. Before thawing the oocyte, the patient attempted pregnancy naturally for 12 months, followed by two unsuccessful clomiphene citrate ovulation induction cycles with intrauterine insemination and one fresh IVF cycle resulting in a chromosomally abnormal twin gestation that aborted. CONCLUSION(S): Although oocyte cryopreservation is still labeled an experimental procedure, this case demonstrates that oocyte cryopreservation used for electively deferred reproduction can subsequently serve in the treatment for secondary infertility when the patient becomes her own oocyte donor
— id: 107365, year: 2010, vol: 93, page: 2413.e7, stat: Journal Article,

Surviving childhood and reproductive-age malignancy: effects on fertility and future parenthood
Knopman, Jaime M; Papadopoulos, Esperenza B; Grifo, James A; Fino, M Elizabeth; Noyes, Nicole
2010 May;11(5):490-498, Lancet oncology
Annually, more than 50 000 cancer diagnoses are made in the USA in patients under the age of 35 years. Despite this staggering statistic, medical advancements have substantially improved survival rates. Thus, for both male and female patients with cancer, quality-of-life issues, such as fertility preservation and parenthood, have become an essential component of treatment. Unfortunately, many of the treatments to eradicate malignant processes can also compromise reproductive function. In these cases, fertility preservation should be discussed and initiated with early treatment planning, to allow the best chance for future parenthood, when appropriate. The effects of cancer and cancer treatments on fertility and future parenthood, including health risks for patients, their gametes, and offspring are discussed
— id: 107364, year: 2010, vol: 11, page: 490, stat: Journal Article,

Fate of cryopreserved donor embryos
Knopman, Jaime M; Talebian, Sheeva; Berkeley, Alan S; Grifo, James A; Noyes, Nicole; Licciardi, Frederick
2010 Oct;94(5):1689-1692, Fertility & sterility
OBJECTIVE: To review a center's experience with cryopreserved embryos generated from donor eggs and to analyze their long-term disposition. DESIGN: Retrospective analysis of donor egg cycles with cryopreserved embryos. SETTING: University-based IVF program. PATIENT(S): Eight hundred twenty-nine women undergoing oocyte donation. INTERVENTION(S): N/A. MAIN OUTCOME MEASURE(S): Factors affecting the decision regarding disposition of donor frozen embryo transfer (dFET) and the association between fresh and dFET cycles. RESULT(S): From January 2000 to December 2004, donor egg recipients underwent 829 fresh embryo transfer cycles that resulted in a 54% live birth rate. Of the 444 recipients who delivered, 177 (40%) also cryopreserved embryos at transfer; however, only 37 (21%) returned for a dFET by August 2009 and only 18 women had children from fresh and frozen transfers. In contrast, 128 of the 385 recipients who failed the fresh transfer (33%) cryopreserved embryos and 111 (87%) returned for a dFET. Of these, 44 had children from the dFET. Frozen cycle success rates between these recipient groups did not depend on fresh cycle outcome or prior parity. CONCLUSION(S): Donor oocyte recipients often initiate treatment with a desire to cryopreserve embryos for future use and family expansion. However, our data demonstrates that most recipients with a child from the fresh transfer do not return to use their cryopreserved embryos. Although fresh transfer success correlated with embryo disposition, it did not correlate with the outcome of thawed embryo transfer
— id: 138166, year: 2010, vol: 94, page: 1689, stat: Journal Article,

Comparison of pregnancy outcomes in anonymous shared versus exclusive donor oocyte in vitro fertilization cycles
Mullin, Christine M; Fino, M Elizabeth; Talebian, Sheeva; Keegan, Debbra; Grifo, Jamie A; Licciardi, Frederick
2010 Feb;93(2):574-578, Fertility & sterility
OBJECTIVE: To determine whether there is a difference in pregnancy outcomes between women undergoing a shared versus exclusive donor oocyte cycle. DESIGN: Retrospective analysis. SETTING: University IVF center. PATIENT(S): Women undergoing either a shared (n=656 cycles), exclusive (n=225 cycles), or shared converted to exclusive (n=22 cycles) donor oocyte cycle from January 2000-December 2005. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Number of eggs retrieved, eggs fertilized, embryos transferred, embryos cryopreserved, clinical pregnancy rates (PR), live birth rates, spontaneous abortion rates. RESULT(S): Pregnancy outcomes in 656 shared cycles were compared with 225 exclusive cycles and 22 shared converted to exclusive donor oocyte cycles. Overall, there was no difference in the clinical PR among the three groups; however, the exclusive group did have a significantly greater number of embryos cryopreserved and this event occurred more frequently in such a cycle. CONCLUSION(S): Women undergoing a donor oocyte IVF cycle can choose to share the donor's oocytes with another recipient without compromising their PR; however, the probability of cryopreservation in such a shared donor oocyte cycle is significantly reduced. Therefore, the recipient must weigh the financial burden of an exclusive cycle with the desires for cryopreservation in an IVF cycle
— id: 114626, year: 2010, vol: 93, page: 574, stat: Journal Article,

Comparison of pregnancy outcomes in elective single blastocyst transfer versus double blastocyst transfer stratified by age
Mullin, Christine M; Fino, M Elizabeth; Talebian, Sheeva; Krey, Lewis C; Licciardi, Frederick; Grifo, Jamie A
2010 Apr;93(6):1837-1843, Fertility & sterility
OBJECTIVE: To determine whether there is a difference in pregnancy outcomes, stratified by age, between women undergoing elective single blastocyst transfer (eSBT) versus those undergoing double blastocyst transfer (2BT). DESIGN: Retrospective analysis. SETTING: University IVF center. PATIENT(S): A total of 1,141 nondonor IVF cycles in women aged <40 years from January 2004-March 2007. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Eggs retrieved, embryos cryopreserved, implantation rates, clinical pregnancy rates (PR), live birth rates (LBR), spontaneous abortion rates (SAB). RESULT(S): Pregnancy outcomes in 52 cycles of women <40 years of age who underwent eSBT were compared with 1,086 cycles of women who underwent 2BT in fresh IVF cycles from January 2004-March 2007. Overall, the eSBT was associated with a statistically significant 92% reduction in the twinning rate (from 25%-2%) while maintaining a high clinical PR (63% in the eSBT group vs. 61% in the 2BT group). CONCLUSION(S): Women who are <40 years of age undergoing nondonor fresh IVF cycles can electively choose to transfer a single blastocyst for the purpose of significantly reducing their risk of multiples without compromising their PR
— id: 95765, year: 2010, vol: 93, page: 1837, stat: Journal Article,

Quality-of-life (QOL) assessment at time of fertility preservation (FP) counseling in female cancer patients: Results of a university-based registry at two years
Noyes N.; Reh A.; Mullin C.; Fino M.E.; Grifo J.A.
2010 ;28(15 SUPPL 1):?-?, Journal of clinical oncology
Background: Objectives - 1) Establish a prospective registry with long-term follow-up of female cancer patients (CP) who presented for initial consultation for FP; 2) Introduce the FACT-B QOL survey for novel use in a FP population. Methods: All consenting female CP completed the registry survey/FACT-B and agreed to yearly follow-up. Results: From 3/2008 to 1/2010, 22 CP enrolled in the registry, 2 declined, and 1 was lost to follow-up. Cancer diagnoses included 6 cervical, 4 breast, 3 endometrial, 2 ovarian, 2 Hodgkin's, 1 childhood neuroblastoma + renal cell carcinoma, 1 Ewing's sarcoma, 1 thymic carcinoma, 1 AML, 1 papillary mesothelioma. The average age at enrollment was 33.1+/-5 (range 23 - 44) y. Sixteen patients sought FP with oocyte (OC) and/or embryo cryopreservation (EC), and 5 underwent in vitro fertilization (IVF). 15/22 were referred from their medical or surgical oncologists. Thirteen patients sought treatment prior to cancer surgery and/or chemo and began FP treatment an average of 12 (range 1 - 22) days from initial consult. Twenty patients completed 30 cycles; 14 used OC and/or EC, 5 IVF and 1 oocyte donation (OD), of which 7 patients underwent embryo transfers resulting in 1 delivery, 1 ongoing (in 8^th month) and 1 miscarried pregnancy. One year follow-up is ongoing. By survery, most patients (12/22) felt having a child was most important in their life (scale 1 - 7; mean 6.1), and 14/22 were most concerned with the impact of their cancer treatment on fertility (mean 6.1). IVF and OD were the most and least preferred treatments, respectively. Recognizing the limited data on the long-term risks for FP patients, most (14/22) were unsure regarding the risk they would undertake to pursue fertility treatment. Most patients reported high baseline QOL scores across all categories of physical, social, emotional, and functional well being. Conclusions: Fertility after cancer remains a significant issue, yet most patients are unsure of their risk tolerance. We introduce the FACT-B QOL survey for novel use in a FP population and aim to determine the long-term safety, effectiveness, and QOL impact of FP treatment in female CP
— id: 112441, year: 2010, vol: 28, page: ?, stat: Journal Article,

Oocyte cryopreservation outcomes including pre-cryopreservation and post-thaw meiotic spindle evaluation following slow cooling and vitrification of human oocytes
Noyes, Nicole; Knopman, Jaime; Labella, Patty; McCaffrey, Caroline; Clark-Williams, Melicia; Grifo, Jamie
2010 Nov;94(6):2078-2082, Fertility & sterility
OBJECTIVE: To report our oocyte cryopreservation (OC) outcomes including meiotic spindle (MS) evaluation of metaphase II (MII) oocytes destined for OC and thaw. DESIGN: Retrospective. SETTING: University-based infertility center. PATIENT(S): Women attempting pregnancy using cryopreserved oocytes. INTERVENTION(S): OC, MS evaluation. MAIN OUTCOME MEASURE(S): Survival, two pronuclear (2PN) fertilization, achieving embryo quality suitable for transfer or refreezing, blastocyst formation. RESULT(S): Thirty-two OC-thaw cycles resulted in 20 pregnancies, 18 either ongoing or delivered. In 26 cycles, MS evaluation was performed: 262/303 (86%) thawed/recovered oocytes survived, 218/262 (83%) achieved 2PN fertilization, 133/218 (61%) became suitable for day-3 and 122/218 (56%) for day-5 transfer. In total, 58 embryos were transferred resulting in a 62% pregnancy and a 41% implantation rate. Of oocytes evaluated before cryopreservation, 247 (82%) were spindle-positive; 96% of these were also spindle-positive after thawing. Blastocyst formation and suitability for day-5 transfer was achieved more often if a post-thaw spindle was visualized. Of all slow-cooled and vitrified oocytes, a higher percentage of those slow-cooled achieved 2PN fertilization and usability. MS evaluation of oocytes cryopreserved by either method was associated with similar outcomes. CONCLUSION(S): OC outcomes are improving. An MS was almost always exhibited both before cryopreservation and after thawing, suggesting that, with appropriate technique, OC presents minimal harm to the MII oocyte. A meiotic spindle evaluation might help to further OC technology
— id: 114041, year: 2010, vol: 94, page: 2078, stat: Journal Article,

Oocyte cryopreservation: a feasible fertility preservation option for reproductive age cancer survivors
Noyes, Nicole; Labella, Patty Ann; Grifo, James; Knopman, Jaime M
2010 Aug;27(8):495-499, Journal of assisted reproduction & genetics
PURPOSE: To compare oocyte cryopreservation cycles performed in cancer patients to those of infertile women. METHODS: Cancer patients referred for fertility preservation underwent counseling in compliance with the ASRM; those electing oocyte cryopreservation were included. Ovarian stimulation was achieved with injectable gonadotropins and freezing was performed using slow-cooling and vitrification methods. RESULTS: Fifty cancer patients (mean age 31 y) underwent oocyte cryopreservation; adequate ovarian stimulation was achieved in 10 +/- 0.3 days. The outcome from these cycles included a mean peak estradiol of 2,376 pg/ml and an average of 19 oocytes retrieved (15 mature oocytes were cryopreserved/cycle). All patients tolerated ovarian hyperstimulation. There were no significant differences noted between cryopreservation cycles performed in cancer patients and in women without malignancy. CONCLUSIONS: Oocyte cryopreservation appears to be a feasible fertility preservation method for reproductive-age women diagnosed with cancer. This modality is not only effective but also, providing a multidiscipline effort, can be completed in timely fashion
— id: 112546, year: 2010, vol: 27, page: 495, stat: Journal Article,

WHAT IS A NORMAL THYROID STIMULATING HORMONE (TSH) LEVEL? EFFECTS OF STRICTER TSH THRESHOLDS ON PREGNANCY OUTCOMES AFTER IVF
Reh, A.; Danoff, A.; Grifo, J.
2010 SEP ;94(4):S188-S188, Fertility & sterility
— id: 113772, year: 2010, vol: 94, page: S188, stat: Journal Article,

EFFECT OF AUTOIMMUNE THYROID DISEASE (AITD) IN OLDER, EUTHYROID INFERTILE WOMEN UNDERGOING IN VITRO FERTILIZATION (IVF)
Reh, A.; Im, S.; Amarosa, A.; Rolnitzky, L.; Grifo, J.; Danoff, A.
2010 SEP ;94(4):S189-S190, Fertility & sterility
— id: 113773, year: 2010, vol: 94, page: S189, stat: Journal Article,

Optimizing embryo selection with day 5 transfer
Reh, Andrea; Fino, Elizabeth; Krey, Lewis; Berkeley, Alan; Noyes, Nicole; Grifo, James
2010 Feb;93(2):609-615, Fertility & sterility
OBJECTIVE: To compare rates of implantation, pregnancy, miscarriage, multiple gestation, and selective reduction between patients undergoing day 5 (d5) and day 3 (d3) ETs. DESIGN: Retrospective cohort study. SETTING: University-based IVF center. PATIENT(S): The first d5 ET cycle of patients 42 years of age from 2003 to 2006 was compared with a historical control of first cycle d3 ET patients 42 years of age from 1996 to 1999 who would have met current d5 ET criteria. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Rates of implantation, clinical pregnancy, miscarriage, live birth, high order multiple pregnancy (HOMP), and selective reduction. RESULT(S): D5 ET patients had higher implantation rates (39% vs. 30%), with no difference in the no-transfer rate. D5 ET patients had lower rates of HOMP (2.5% vs. 11%) and HOMP delivery (0.7% vs. 3.5%), multiple pregnancy (27% vs. 33%), multiple delivery (19% vs. 26%), and twin delivery (18% vs. 23%). There were fewer selective reductions of HOMP with d5 ET (1.7% vs. 3.8%). CONCLUSION(S): Extended culture improves embryo selection through increased implantation, facilitating fewer embryos per transfer, which lowers multiple gestation rates and the need for HOMP reduction
— id: 99204, year: 2010, vol: 93, page: 609, stat: Journal Article,

What is a normal thyroid-stimulating hormone (TSH) level? Effects of stricter TSH thresholds on pregnancy outcomes after in vitro fertilization
Reh, Andrea; Grifo, James; Danoff, Ann
2010 Dec;94(7):2920-2922, Fertility & sterility
Using a thyroid-stimulating hormone (TSH) cutoff of 2.5 mIU/L or 4.5 mIU/L, no differences in the rates of clinical pregnancy, delivery, or miscarriage were observed in this large, retrospective cohort study of first-cycle IVF patients from 2005 through 2008, after controlling for age. Although lowering the TSH threshold to 2.5 mIU/L would result in a nearly fivefold increase in the number of women being classified as hypothyroid, the lack of differences in maternal clinical outcomes must be considered in the current controversy regarding the relative merits of lowering the upper limit of normal of TSH
— id: 149782, year: 2010, vol: 94, page: 2920, stat: Journal Article,

ESTRADIOL (E2), PROGESTERONE (P), AND HUMAN CHORIONIC GONADOTROPIN (hCG) AS PREDICTORS OF PREGNANCY OUTCOME IN IN-VITRO FERTILIZATION (IVF)
Weinerman, R. S.; Mullin, C.; Grifo, J. A.
2010 SEP ;94(4):S259-S259, Fertility & sterility
— id: 113774, year: 2010, vol: 94, page: S259, stat: Journal Article,

ARE WE UNDERESTIMATING THE PREVALENCE OF ANEUPLOIDY- RELATED MISCARRIAGES? A DESCRIPTION OF CYTOGENETIC RESULTS FROM PRODUCTS OF CONCEPTION (POC) AFTER DILATION AND CURETTAGE (D&C)
Werner, M. D.; Reh, A.; Perle, M. A.; Grifo, J.
2010 SEP ;94(4):S42-S42, Fertility & sterility
— id: 113764, year: 2010, vol: 94, page: S42, stat: Journal Article,

Novel highly efficient generation of disease-specific human embryonic stem cells from genetically abnormal embryos
Hansis C.; Rice C.E.; Grifo J.A.; Lehmann R.
2009 ;92(3 SUPPL 1):S86-S86, Fertility & sterility
OBJECTIVE: Since many frequent, fatal, and incurable diseases do not have an appropriate disease model and with attainable embryos being scarce, with IRB approval we attempted to generate novel disease-specific human embryonic stem (hES) cells with new protocols for more efficient derivation, maintenance, and differentiation. DESIGN: Experimental. MATERIALS AND METHODS: Zona-free human blastocysts (n=14) previously assessed by preimplantation genetic diagnosis (PGD) for genetic conditions were transferred onto feeder cells and cultured in DMEM-based media. Pieces of the colonies (n=4) were frozen by liquid nitrogen vitrification with cryoprotectants propylene glycol, DMSO, and acetamide and subsequently thawed. Differentiation of hES cells was achieved by colony overgrowth or embryoid body formation. Embryonic and control cells were subjected to marker gene and protein analysis for pluripotency and differentiation by reverse transcription PCR and immunofluorescence, respectively. RESULTS: Colonies (n=10; 71.4% of transferred blastocysts) could be established and maintained which showed the typical morphological features of hES cells such as compact colony formation. Colonies were derived from affected embryos (one each of cystic fibrosis, trisomy X, 18, 21, 22, and Tay-Sachs disease), from embryos tested inconclusively (one of cystic fibrosis and three of Tay-Sachs disease), and from three normal control embryos. Marker gene and protein expression as well as growth pattern analysis suggest that the colony cells retain their undifferentiated state in culture as well as after vitrification and thawing and that they can be differentiated into a variety of cell types, including the tissues most affected by the conditions. CONCLUSIONS: These newly established protocols for the derivation, maintenance, and differentiation of novel disease-specific hES cell lines should enable the efficient generation of new disease models. This will provide new tools to study diseases as well as to develop new therapeutic approaches
— id: 127243, year: 2009, vol: 92, page: S86, stat: Journal Article,

Women with cancer undergoing ART for fertility preservation: a cohort study of their response to exogenous gonadotropins
Knopman, Jaime M; Noyes, Nicole; Talebian, Sheeva; Krey, Lewis C; Grifo, James A; Licciardi, Frederick
2009 Apr;91(4 Suppl):1476-1478, Fertility & sterility
Cancer patients produce similar numbers of oocytes after ovarian hyperstimulation compared with age-matched infertile controls, suggesting that malignancy does not adversely affect ovarian response
— id: 90883, year: 2009, vol: 91, page: 1476, stat: Journal Article,

Outcomes of medically-indicated (MED) oocyte cryopreservation (OC) cycles performed for fertility preservation (FP) compare favorably to oc cycles of infertile women who have completed a thaw cycle
Noyes N.; Knopman J.; Labella P.; Werner M.; McCaffrey C.; Grifo J.A.
2009 ;92(3 SUPPL 1):S224-S224, Fertility & sterility
OBJECTIVE: As more reproductive-age women are diagnosed with and survive cancer, OC use is increasing as a means of FP, particularly in those without a male partner. We compared outcomes of medically-indicated (MED) OC cycles to those of infertile women completing an OC thaw cycle (THAW), including their meiotic spindle evaluation (MSE). All MII oocytes cryopreserved in our lab undergo MSE 2 h post-harvest. DESIGN: Retrospective. MATERIALS AND METHODS: 31 MED (520 retrieved eggs) and 28 THAW (512 retrieved eggs) OC cycles were reviewed. Tumors in the MED group included 13 GYN, 10 breast, 3 leukemia, 2 colon, 2 CNS and 1 sarcoma. All patients underwent standard ovarian hyperstimulation followed by retrieval, MSE 2 h post-harvest, then OC of all MII oocytes. Both slow freezing and vitrification were used. 4 MED pts froze ¹?₂ as zygotes. RESULTS: In the THAW cycles, there were 16 (57%) pregnancies: 12 women delivered 17 liveborns (5 twins), 3 have ongoing gestations and 1 miscarried. 90% of eggs survived and there was a 94% pre-cryo to post-THAW MSE correlation. 2 MED thaw cycles resulted in 1 pregnancy. Cycle comparisons for THAW vs. MED are shown in the Table. (Table presented). CONCLUSIONS: The majority of oocytes in both groups were mature (MII) and exhibited a spindle. Women diagnosed with cancer can achieve adequate ovarian stimulation, egg production and MSE despite limited time to complete OC treatment and multiple outside stressors, including their underlying disease and its management. Based on our preliminary THAW data, we believe cancer pts should have a reasonable chance for pregnancy using their frozen eggs
— id: 127248, year: 2009, vol: 92, page: S224, stat: Journal Article,

Non-invasive pre-cryo and post-thaw meiotic spindle (MS) evaluation of human oocytes cyropreserved (OC) by either slow freezing or vitrification: A value or time wasted?
Noyes N.; Labella P.; McCaffrey C.; Clark-Williams M.; Novetsky A.P.; Grifo J.A.
2009 ;92(3 SUPPL 1):S195-S196, Fertility & sterility
OBJECTIVE: The MS is critical to chromosome segregation and embryo competence. Today, polarized light microscopy (PM) affords non-invasive MS evaluation (MSE). We perform PM 2 h post-harvest and 1 h post-thaw. Here, we appraise its value. DESIGN: Retrospective. MATERIALS AND METHODS: 28 OC cycles with thaw were assessed; 1/2 eggs were slow frozen and 1/2 vitrified (vit); pt's eggs were split between methods. MSE was done in the last 23 cycles. Outcome measures included 2PN fertilization (2PN-F), embryo quality suitable for ET, blast formation rate (BFR), and embryo usability (transferred or refrozen). RESULTS: All eggs were from women <38 (mean 31) yowith normal D3 FSH/E2. Of 28 cycles, 57% had pregnancy; 12 delivered 17 liveborns, 3 are ongoing and 1 aborted. In the last 25 cycles, 286 MII eggs were thawed after cryo for a mean of 3 mos; pre-cryo, 83% were MS positive (SP+). 255 (89%) survived; 89% were SP+ post-thaw, a 94% pre-cryo and post-thaw MSE correlation. 211 (83%) ICSI'd eggs achieved 2PN-F. 2.3 embryos were transferred/cycle (30 slow & 22 vit). Pre-cryo MSE: no difference was noted in egg survival or 2PN-F. 65% of SP+ vs. 44% of MS negative (SP-) zygotes became embryos suitable for ET D3 post-thaw (p=.02). No difference was noted in the BFR or % of embryos suitable for D5 ET, but 86% of embryos transferred or refrozen were from SP+ eggs. Post-thaw MSE: no difference was noted in the % of embryos suitable for ET D3 post-thaw or BFR. A significantly higher % of embryos were suitable for ET D5 post-thaw in the SP+ (57%) vs. SP- (29%) groups (p=.03) and 95% of transferred or refrozen embryos were from SP+ oocytes. When comparing MSE data for slow vs. vit eggs, no differences were noted. CONCLUSIONS: Most retrieved oocytes (slow and vit) exhibited a MS pre & post-thaw and most transferred or refrozen embryos were from SP+ oocytes. OC does not appear to damage the MS and viewing it may correlate with oocyte competence. Because MSE may improve efficiency of gamete selection, it should be considered to advance the field of OC
— id: 127247, year: 2009, vol: 92, page: S195, stat: Journal Article,

Ectopic pregnancy rates after in vitro fertilization: a look at the donor egg population
Rosman, Elana R; Keegan, Debbra A; Krey, Lewis; Liu, Mengling; Licciardi, Frederick; Grifo, Jamie A
2009 Nov;92(5):1791-1793, Fertility & sterility
In an 8-year review of ectopic pregnancy (EP) rates in donor egg recipients and standard patients undergoing in vitro fertilization-embryo transfer (IVF-ET) at a large university-based program, we report an EP rate of 0.6% in donor egg recipients and 0.9% in standard IVF patients, a difference that is not statistically significant. Donor egg recipients were found to have a significantly lower incidence of tubal disease compared with standard IVF patients; however, tubal disease was not found to be an independent risk factor for EP in our practice, perhaps owing to aggressive management of tubal disease
— id: 100679, year: 2009, vol: 92, page: 1791, stat: Journal Article,

Laboratory evaluation in oocyte cryopreservation (OC) cycles suggests retrieved oocytes are comparable whether frozen for medical indications (MED), deferred reproduction (DR) or oocyte donation (OD)
Werner M.D.; Labella P.A.; Grifo J.A.; Kramer Y.; Reh A.; Noyes N.
2009 ;92(3 SUPPL 1):S188-S188, Fertility & sterility
OBJECTIVE: The use of OC is increasing. We know the human oocyte meiotic spindle (MS) is crucial to chromosome segregation and embryo development while the zona pellucida (ZP) aids in sperm binding and fertilization. Today, polarized microscopy (PM) affords non-invasive MS and ZP evaluation. All MII oocytes cryopreserved in our lab undergo PM MS and ZP assessment 2 h post-harvest. We compared OC cycle outcome in cases performed for MED vs. DR vs. OD. DESIGN: Retrospective. MATERIALS AND METHODS: All women were <40 y at time of OC. Tumors in the MED group included 13 GYN, 10 breast, 3 leukemia, 2 colon, 2 CNS & 1 sarcoma. All patients underwent standard ovarian stimulation followed by retrieval, PM for MS and ZP, then OC of MII oocytes. RESULTS: Cycle outcome is shown in the Table. (Table presented). CONCLUSIONS: The majority of retrieved oocytes in all groups were MII. Women diagnosed with malignancy achieved adequate ovarian stimulation, egg production and a similar number of spindle positive eggs compared to DR and OD patients, despite limited time for treatment and multiple outside stressors, including underlying disease. MS & ZP measurements were clinically comparable whether eggs were frozen for cancer, DR or OD
— id: 127246, year: 2009, vol: 92, page: S188, stat: Journal Article,

Embryo biopsy: the fate of abnormal pronuclear embryos
Noyes, Nicole; Fino, M Elizabeth; Krey, Lewis; McCaffrey, Caroline; Adler, Alexis; Grifo, James
2008 Dec;17(6):782-788, Reproductive biomedicine online
This study assessed 1908 embryos, including those with abnormal numbers of pronuclei, in IVF cycles from July 2001 to December 2006 in which preimplantation genetic screening (PGS) was performed on day 3 post-retrieval and 'euploid' embryos transferred the following day. PGS-intracytoplasmic sperm injection and PGS-translocation cycles were excluded. At 18 h post-insemination, the zygote distribution was 19% 0PN, 4% 1PN, 69% 2PN and 8% 3PN. No pregnancy occurred following 0PN or 1PN embryo transfers. A retrospective, blinded morphological ranking of all embryos on day 3 was performed and the results compared with PGS; no 0PN or 1PN embryo would have been chosen for transfer based on morphological superiority alone. Blastocyst formation occurred in 1PN embryos (29%) but not in 0PN embryos when evaluated on day 5. Euploid karyotypes were reported for biopsies of 0PN (3%), 1PN (5%) and 2PN (19%) embryos (P = 0.015, 1PN versus 2PN). A Y chromosome was observed in 0PN (17%) and 1PN (32%) embryos; surprisingly, 91% of these Y chromosome-bearing embryos were aneuploid. Many different meiotic and fertilization errors can result in 0PN or 1PN zygotes; these results indicate that the resultant embryos should not be transferred, especially when normally fertilized embryos are available
— id: 96879, year: 2008, vol: 17, page: 782, stat: Journal Article,

Using certain commercially available glassbottomed dish with paraffin oil impacts the embryonic development of oocytes
Chang, HC; Grifo, J; McCaffrey, C; Krey, LC; Noyes, N
2007 SEP ;88(3):S310-S311, Fertility & sterility
— id: 74427, year: 2007, vol: 88, page: S310, stat: Journal Article,

Multicentre trial of preimplantation genetic screening reported in the New England Journal of Medicine: an in-depth look at the findings
Cohen, Jacques; Grifo, James A
2007 Oct;15(4):365-366, Reproductive biomedicine online
A randomized clinical trial of 406 patients with advanced maternal age by Mastenbroek and co-workers recently published in the New England Journal of Medicine showed a significant decrease in pregnancy outcome after preimplantation genetic screening (PGS). It is our opinion that this study suffers from a number of insurmountable inaccuracies and that these are either a direct consequence of the inexperience of the team or of a general disregard of vital guidelines reported in the literature. Most importantly, the authors show that in their hands embryo biopsy may affect as many as half the embryos. The error rate was not presented, shedding doubt on the authors' abilities to reliably diagnose the biopsied cells. An evaluation of the study indicates that poor biopsy technique, sub standard fixation and FISH methods, poor IVF outcomes and inappropriate patient selection are the cause of the discouraging results obtained by these authors rather than problems inherent to PGS
— id: 96502, year: 2007, vol: 15, page: 365, stat: Journal Article,

Does embryo morphology predict ploidy? An update on the usefulness of PGD testing for aneuploidy
Fino, E; Talebian, S; Grifo, J; McCaffrey, C; Adler, A; Noyes, N
2007 SEP ;88(3):S230-S231, Fertility & sterility
— id: 74420, year: 2007, vol: 88, page: S230, stat: Journal Article,

Is what we clearly see really so obvious? Ultrasonography and transcervical embryo transfer--a review
Flisser, Eric; Grifo, Jamie A
2007 Jan;87(1):1-5, Fertility & sterility
OBJECTIVE: To critically review the role of ultrasound-guided embryo transfer (ET) and its influence on the outcome of in vitro fertilization (IVF). DESIGN: Medline review of published manuscripts. RESULT(S): Studies evaluating the role of ultrasound-assisted ET have had mixed results, and although meta-analysis of prospective trials suggests an improvement in outcome, limitations in study design may overstate the effect of ultrasonography. Other ET techniques may eliminate the advantages provided by ultrasonography, limiting its benefit to specific clinical scenarios. However, because no trial has demonstrated an adverse effect and because cases that may benefit from its use often cannot be predicted reliably, the routine application of ultrasonography can be justified
— id: 70091, year: 2007, vol: 87, page: 1, stat: Journal Article,

The role of oocyte donation,in expanding the natural family
Gowda, M; Talebian, S; Noyes, N; Berkeley, AS; Grifo, JA; Licciardi, F
2007 SEP ;88(3):S257-S257, Fertility & sterility
— id: 74423, year: 2007, vol: 88, page: S257, stat: Journal Article,

Ten-year experience with preimplantation genetic diagnosis (PGD) at the New York University School of Medicine Fertility Center
Grifo, J; Talebian, S; Keegan, D; Krey, L; Adler, A; Berkeley, A
2007 Oct;88(4):978-981, Fertility & sterility
We describe our experience of over 300 cycles of preimplantation genetic diagnosis (PGD) and report clinical pregnancy rates (35%-67%) that support using this technology to screen for genetic disorders and chromosomal abnormalities. In clinical practice for over ten years, PGD offers couples the earliest form of genetic screening and may help improve ongoing pregnancy rates in poor-prognosis patients
— id: 74660, year: 2007, vol: 88, page: 978, stat: Journal Article,

Programmatic implementation of blastocyst transfer in a university-based in vitro fertilization clinic: maximizing pregnancy rates and minimizing triplet rates
Grifo, James A; Flisser, Eric; Adler, Alexis; McCaffrey, Caroline; Krey, Lewis C; Licciardi, Frederick; Noyes, Nicole; Kump, Lisa M; Berkeley, Alan S
2007 Aug;88(2):294-300, Fertility & sterility
To assess whether the use of extended embryo culture can reduce the incidence of high-order multiple gestations, a retrospective analysis of 7,418 fresh ETs performed in a university-based IVF clinic from 1997-2003 was conducted, comparing program results before and after institution of a protocol to select patients for extended culture of in vitro fertilized embryos. The incidence of triplet pregnancies was significantly reduced in patients at highest risk for high-order multiple gestations, i.e., those at <35 years of age (16.8% versus 6.8%), those at 35-37 years of age (13.0% versus 5.6%), and recipients of donated oocytes (11.2% versus 4.5%)
— id: 73928, year: 2007, vol: 88, page: 294, stat: Journal Article,

A study of sperm motility as a predictor of pregnancy outcome, and analysis of NYUPIVF pregnancy outcomes
Immerman, Sara; Moses, Nicole; Grifo, Jaime
2007 ;1:20-20, Probe: the publication of research on biomedical endeavors
— id: 75317, year: 2007, vol: 1, page: 20, stat: Journal Article,

Isolated teratozoospermia does not affect in vitro fertilization outcome and is not an indication for intracytoplasmic sperm injection
Keegan, Brian Robert; Barton, Sara; Sanchez, Xavier; Berkeley, Alan S; Krey, Lewis C; Grifo, Jamie
2007 Dec;88(6):1583-1588, Fertility & sterility
OBJECTIVE: To reevaluate clinical management of isolated teratozoospermia, in couples initiating IVF. DESIGN: Retrospective analysis of fertility indices in 535 cycles. SETTING: A large, university-based fertility center. PATIENT(S): Consecutive couples (n = 495) who had a semen analysis using Kruger/Tyberberg strict criteria at our center within 12 months before undergoing their first and/or second IVF cycle in 2002-2004 with >2 million postwash, motile sperm on the day of egg retrieval. INTERVENTION(S): Eggs were fertilized either by conventional IVF or ICSI. Semen analysis and gamete/embryo manipulation was standardized in all cases. MAIN OUTCOME MEASURE(S): Fertilization, fertilization failure, pregnancy, and live birth rates. RESULT(S): There was no statistical difference in fertilization, fertilization failure, pregnancy, and live birth rates in the first or second IVF cycle when comparing couples with isolated teratozoospermia (<5% normal morphology) to those with a normal semen analysis. Furthermore, no improvement in these outcomes was noted when ICSI was used to treat these teratozoospermic couples. CONCLUSION(S): Because isolated teratozoospermia generally does not impact on the major indices of IVF, these patients need not be subjected to the unnecessary cost and potential risks of ICSI. Future studies, however, should focus on different sperm morphologic and biochemical parameters to determine if they are important for clinical management in IVF
— id: 75762, year: 2007, vol: 88, page: 1583, stat: Journal Article,

Minimizing the risk of high order multiples without compromising pregnancy rates: one center's 5-year experience with ovulation induction
Keegan, DA; Edenfield, A; Krey, LC; Grifo, J
2007 SEP ;88(3):S255-S255, Fertility & sterility
— id: 74421, year: 2007, vol: 88, page: S255, stat: Journal Article,

Embryo performance in a day 5 extended culture protocol predicts IVF outcome better than age and may be helpful in preventing multiple gestation
Keegan, DA; Talebian, S; Fino, E; Krey, LC; Berkeley, AS; Grifo, JA
2007 SEP ;88(3):S323-S323, Fertility & sterility
— id: 74431, year: 2007, vol: 88, page: S323, stat: Journal Article,

Availability of good quality blastocysts for cryopreservation (CRYO) on day 5 (d5) of a fresh embryo transfer (ET) identifies candidates for single blastocyst transfer (SBT)
Keegan, DA; Talebian, S; Fino, E; Krey, LC; Grifo, JA; Berkeley, AS
2007 SEP ;88(3):S316-S316, Fertility & sterility
— id: 74430, year: 2007, vol: 88, page: S316, stat: Journal Article,

Low ectopic pregnancy rates after in vitro fertilization: do practice habits matter?
Keegan, Debbra A; Morelli, Sara S; Noyes, Nicole; Flisser, Eric D; Berkeley, Alan S; Grifo, Jamie A
2007 Sep;88(3):734-736, Fertility & sterility
In a 6-year review of ectopic pregnancies (EPs) after fresh and frozen embryo transfers in IVF cycles conducted at a large university-based program, we report an overall 0.9% rate of EP that seems to have increased with the programmatic shift to routine blastocyst transfer, but remains lower than nationally reported rates. Aggressive management of tubal disease may contribute to low rates of EP, whereas blastocyst transfer may increase the rate
— id: 71127, year: 2007, vol: 88, page: 734, stat: Journal Article,

Heterotopic abdominal pregnancy following two-blastocyst embryo transfer
Knopman, Jaime M; Talebian, Sheeva; Keegan, Debbra A; Grifo, James A
2007 Nov;88(5):1437.e13-1437.e15, Fertility & sterility
OBJECTIVE: To report a case of a heterotopic primary abdominal pregnancy after two-blastocyst IVF-ET. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A woman with a heterotopic abdominal pregnancy after IVF-ET. INTERVENTION(S): Pituitary down-regulation with luteal antagon, ovulation induction with menotropins, IVF-ET, progesterone in oil for luteal support, dilation and curettage for missed abortion, laparoscopy, and resection of abdominal gestation. MAIN OUTCOME MEASURE(S): Human chorionic gonadotropin levels, pelvic ultrasound examinations, and laparoscopic and pathologic findings. RESULT(S): A heterotopic abdominal pregnancy occurred after a two-blastocyst IVF-ET. The concurrent intrauterine gestation resulted in a miscarriage. CONCLUSION(S): The number of embryos transferred has been identified as a powerful risk factor for heterotopic pregnancy; however, heterotopic pregnancy can occur following a two-embryo, blastocyst stage transfer
— id: 75357, year: 2007, vol: 88, page: 1437.e13, stat: Journal Article,

Do patients with successful donor embryo cycles have children from their supernumery cryopreserved embryos?
Knopman, JM; Talebian, S; Krey, LC; Berkeley, AS; Grifo, JA; Licciardi, F
2007 SEP ;88(3):S258-S258, Fertility & sterility
— id: 74424, year: 2007, vol: 88, page: S258, stat: Journal Article,

The fate of cryopreserved donor embryos
Knopman, JM; Talebian, S; Noyes, N; Krey, LC; Grifo, JA; Licciardi, F
2007 SEP ;88(3):S256-S257, Fertility & sterility
— id: 74422, year: 2007, vol: 88, page: S256, stat: Journal Article,

Polycystic ovarian syndrome (Pcos) significantly alters follicular fluid molecular structure
Levine BA; Flisser E; Katz J; Levitz M; Noyes N; Krey LC; Grifo JA
2007 ;1:19-19, Probe: the publication of research on biomedical endeavors
— id: 75316, year: 2007, vol: 1, page: 19, stat: Journal Article,

Equivalent viability in mouse MII oocytes exposed to choline-based and sodium-based vitrification procedures
Liu, H; Krey, LC; Grifo, JA
2007 SEP ;88(3):S353-S354, Fertility & sterility
— id: 74434, year: 2007, vol: 88, page: S353, stat: Journal Article,

Mouse MII oocytes can tolerate a wide range of exposure time to high concentration of cryoprotectants through vitrification procedure
Liu, H; Krey, LC; Grifo, JA
2007 SEP ;88(3):S356-S356, Fertility & sterility
— id: 74435, year: 2007, vol: 88, page: S356, stat: Journal Article,

Going towards single embryo transfer
McCaffrey, C; Adler, A; Krey, L; Noyes, N; Berkeley, A; Grifo, J
2007 SEP ;88(3):S331-S331, Fertility & sterility
— id: 74432, year: 2007, vol: 88, page: S331, stat: Journal Article,

Visualization of meiotic spindle predicts usefulness of female gametes in an oocyte cryopreservation (OC) program
McCaffrey, C; Fino, E; Grifo, JA
2007 SEP ;88(3):S335-S335, Fertility & sterility
— id: 74433, year: 2007, vol: 88, page: S335, stat: Journal Article,

Comparison of pregnancy outcomes in anonymous shared vs. exclusive donor oocyte in vitro fertilization (IVF) cycles
Mullin, CM; Talebian, S; Keegan, D; Fine, ME; Grifo, JA; Licciardi, F
2007 SEP ;88(3):S132-S132, Fertility & sterility
— id: 74418, year: 2007, vol: 88, page: S132, stat: Journal Article,

Embryo biopsy the fate of biopsied abnormal pronluclear embryos
Noyes, N; Fino, E; McCaffrey, C; Adler, A; Krey, L; Grifo, J
2007 SEP ;88(3):S29-S30, Fertility & sterility
— id: 74414, year: 2007, vol: 88, page: S29, stat: Journal Article,

Microsort (R) processed sperm requires ICSI; Embryo selection is enhanced by PGD for aneuploidy when used for gender selection in sex-linked disorders
Noyes, N; Melander, B; Adler, A; Grifo, J; Fino, E; Krey, LC
2007 SEP ;88(3):S38-S38, Fertility & sterility
— id: 74415, year: 2007, vol: 88, page: S38, stat: Journal Article,

Day 5 versus day 6 blastocyst cryopreservation
Adler, A; Dejesus, E; Flisser, E; Berkeley, A; Grifo, J; Krey, L
2006 SEP ;86(1-2):S257-S257, Fertility & sterility
— id: 70637, year: 2006, vol: 86, page: S257, stat: Journal Article,

Assessing embryonic potential in a slowly developing cohort by day 6 embryo transfer: Later selection does not compromise pregnancy rates
Adler, A; Keegan, D; Krey, L; Berkeley, A; Grifo, J
2006 SEP ;86(1-2):S257-S257, Fertility & sterility
— id: 70638, year: 2006, vol: 86, page: S257, stat: Journal Article,

Isolated teratozoospermia does not affect initial in vitro fertilization (IVF) outcome and intracytoplasmic sperm injection (ICSI) is not advantageous in this population
Barton, S; Keegan, B; Sanchez, X; Krey, L; Grifo, J
2006 SEP ;86(1-2):S355-S355, Fertility & sterility
— id: 70639, year: 2006, vol: 86, page: S355, stat: Journal Article,

Oct3A and Oct3B immunostaining patterns in human 3PN zygotes and aneuploid embryos diagnosed by preimplantation genetic diagnosis (PGD): Embryonic chromosome status can be independent of developmental competence
Chang, HC; Grifo, J; Labella, P; Ampeloquio, E; Adler, A; Krey, L
2006 SEP ;86(1-2):S220-S221, Fertility & sterility
— id: 70635, year: 2006, vol: 86, page: S220, stat: Journal Article,

Down regulatory effects of retinoid signaling on murine and human embryonic Oct-4 gene expression
Chang, HC; Liu, H; Noyes, N; Meng, L; Krey, L; Grifo, J
2006 SEP ;86(1-2):S219-S220, Fertility & sterility
— id: 70633, year: 2006, vol: 86, page: S219, stat: Journal Article,

Independent and sequential expression of Oct3A and Oct3B transcription factors in human preimplantation embryos
Chang, HC; Noyes, N; Lee, TL; Chin, A; Krey, L; Grifo, J
2006 SEP ;86(1-2):S220-S220, Fertility & sterility
— id: 70634, year: 2006, vol: 86, page: S220, stat: Journal Article,

Transabdominal ultrasound-assisted embryo transfer and pregnancy outcome
Flisser, Eric; Grifo, James A; Krey, Lewis C; Noyes, Nicole
2006 Feb;85(2):353-357, Fertility & sterility
OBJECTIVE: To assess the clinical value of transabdominal ultrasound (TAS)--assisted embryo transfer on outcomes of in vitro fertilization--embryo transfer (IVF-ET) in comparison to the 'clinical touch' method of transcervical embryo transfer by one physician and to determine if transabdominal ultrasound should be applied to all cases of embryo transfer in this practice. DESIGN: A retrospective comparison study. SETTING: A university-based IVF practice. PATIENT(S): Two hundred forty-nine patients who underwent transcervical transfer of fresh embryos created using autologous oocytes performed by the same physician from July 1, 2003, to June 30, 2004. INTERVENTION(S): On selected days, at time of embryo transfer, transabdominal ultrasound was performed to guide catheter placement depth approximately 1 cm from the uterine fundus. MAIN OUTCOME MEASURE(S): The presence of at least one gestational sac on ultrasound was compared in the two study groups; additionally, the clinical pregnancy rate (presence of fetal cardiac activity observed on ultrasound), the ectopic pregnancy rate, the biochemical pregnancy rate, and the implantation rate (number of gestational sacs identified on ultrasound per number of embryos transferred) between groups was evaluated. Characteristics of the two cohorts were analyzed to verify similarities between the treatment and control groups, including age of recipient, prior IVF history, day of transfer (day 3 or day 5 after retrieval), difficulty of transfer, the use of a tenaculum, and the quality and number of embryos transferred. RESULT(S): No statistical difference was seen in the presence or number of gestational sacs following embryo transfer with or without transabdominal ultrasound guidance. CONCLUSION(S): No additional advantage is conferred when using transabdominal ultrasound to perform embryo transfer. In experienced hands, the 'clinical touch' method of embryo transfer yields equivalent results to transabdominal ultrasound-guided embryo placement. However, in patients with a prior history of difficult uterine sounding or embryo transfer, transabdominal ultrasound guidance may still play a role
— id: 64166, year: 2006, vol: 85, page: 353, stat: Journal Article,

Clinical results of an oocyte cryopreservation program
Grifo, JA; Labella, P; Licciardi, F; Chang, H; Lui, H; Noyes, N
2006 SEP ;86(1-2):S127-S127, Fertility & sterility
— id: 70626, year: 2006, vol: 86, page: S127, stat: Journal Article,

Direct matrix assisted laser desorption ionization (MALDI) identification of haptoglobin from culture media of embryos that resulted in a live birth
Keegan, DA; Grifo, JA; Lee, T; Licciardi, F; Naftolin, F; Pevsner, P
2006 SEP ;86(1-2):S219-S219, Fertility & sterility
— id: 70632, year: 2006, vol: 86, page: S219, stat: Journal Article,

Early serum hCG after in vitro fertilization (IVF) differs depending on day of embryo transfer
Keegan, DA; Salas, J; Liu, M; Lukanova, A; Toniolo, P; Grifo, JA
2006 SEP ;86(1-2):S226-S226, Fertility & sterility
— id: 70636, year: 2006, vol: 86, page: S226, stat: Journal Article,

Polycystic ovarian syndrome (PCOs) significantly alters follicular fluid molecular signature
Levine, BA; Flisser, E; Katz, J; Levitz, M; Noyes, N; Krey, LC; Grifo, JA
2006 FEB ;13(2):267A-268A, Journal of the Society for Gynecologic Investigation
— id: 62831, year: 2006, vol: 13, page: 267A, stat: Journal Article,

The viability of mouse oocytes in response to vitrification is dependent on the time of retrieval after HCG stimulation
Liu, H; Krey, L; Grifo, J
2006 SEP ;86(1-2):S203-S203, Fertility & sterility
— id: 70631, year: 2006, vol: 86, page: S203, stat: Journal Article,

Lower culture temperature during vitrification freezing/thawing improves viability of human and mouse oocytes
Liu, H; Labella, P; Chang, H; Noyes, N; Krey, L; Grifo, J
2006 SEP ;86(1-2):S128-S128, Fertility & sterility
— id: 70627, year: 2006, vol: 86, page: S128, stat: Journal Article,

The effect of chelating cytosolic calcium on the viability of vitrified mouse oocytes
Liu, H; Noyes, N; Krey, L; Grifo, J
2006 SEP ;86(1-2):S65-S66, Fertility & sterility
— id: 70625, year: 2006, vol: 86, page: S65, stat: Journal Article,

Presence of meiotic spindle predicts embryo competence following oocyte cryopreservation
Noyes, N; Chang, HC; Liu, H; Labella, P; Meng, L; Grifo, JA
2006 SEP ;86(1-2):S64-S65, Fertility & sterility
— id: 70624, year: 2006, vol: 86, page: S64, stat: Journal Article,

Early serum hCG after in vitro fertilization: Can we predict multiple gestations?
Salas, J; Keegan, DA; Liu, M; Toniolo, P; Grifo, JA
2006 SEP ;86(1-2):S134-S135, Fertility & sterility
— id: 70628, year: 2006, vol: 86, page: S134, stat: Journal Article,

Predictive value of early serum beta-hCG on pregnancy outcome after in vitro fertilization
Salas, J; Keegan, DA; Liu, ML; Lukanova, A; Toniolo, P; Grifo, JA
2006 FEB ;13(2):158A-158A, Journal of the Society for Gynecologic Investigation
— id: 62828, year: 2006, vol: 13, page: 158A, stat: Journal Article,

Vitrification of metaphase II oocytes impacts Oct-4 expression during subsequent preimplantation development
Chang, HC; Grifo, J; Krey, LC
2005 SEP ;84(11):S37-S38, Fertility & sterility
— id: 59560, year: 2005, vol: 84, page: S37, stat: Journal Article,

Developmental incompetency of denuded mouse oocytes undergoing maturation in vitro is ooplasmic in nature and is associated with aberrant Oct-4 expression
Chang, Hung Chi; Liu, Hui; Zhang, John; Grifo, Jamie; Krey, Lewis C
2005 Jul;20(7):1958-1968, Human reproduction
BACKGROUND: Germinal vesicle (GV) oocytes constitute a potential resource but their developmental competence is questionable especially when surrounding cumulus cells are removed. The intercellular factors/mechanisms underlying such poor embryonic competence may originate at a nuclear and/or ooplasmic level. METHODS: Immature or mature oocytes were obtained from three mouse strains following pregnant mare serum gonadotropin (PMSG) or PMSG+ human chorionic gonadotropin (hCG) treatment. Immature oocytes were denuded of cumulus cells prior to in vitro maturation. Pronuclear (PN) transfer was used to examine nuclear-ooplasmic interplay on resultant embryonic development and Oct-4 immuno-staining patterns. RESULTS: Embryos arising from ooplasts of in vivo matured oocytes displayed significant increases in blastocyst formation rates and total blastomere numbers when compared to those created from ooplasts of denuded oocytes. Oct-4 staining was more pronounced and restricted to the inner cell mass (ICM) in blastocysts arising from the ooplasm of in vivo matured zygotes than in those created from denuded oocytes. CONCLUSIONS: Developmental defect(s) appear to develop primarily in the ooplasm of oocytes that are denuded of their cumulus cells prior to in vitro maturation. Such oocytes result in embryos with poor developmental competence. These defects result in anomalies in cell number and Oct-4 expression during the morula-blastocyst developmental transition
— id: 56079, year: 2005, vol: 20, page: 1958, stat: Journal Article,

Reducing the risk of multiple gestations with ovulation induction and intrauterine insemination: One center's experience
Fino, E; Keegan, DA; Noyes, N; Licciardi, F; Berkeley, AS; Grifo, JA
2005 SEP ;84(11):S312-S312, Fertility & sterility
— id: 59570, year: 2005, vol: 84, page: S312, stat: Journal Article,

How good is embryo morphology at predicting chromosomal integrity? When is aneuploidy PGD useful?
Fino, E; Krey, LC; Grifo, JA; McCaffrey, C; Adler, A; Noyes, N
2005 SEP ;84(11):S98-S99, Fertility & sterility
— id: 59563, year: 2005, vol: 84, page: S98, stat: Journal Article,

Surgical correction of uterine septum improves fertility and pregnancy outcome
Fino, E; Noyes, N; Keegan, DA; Grifo, JA; Berkeley, AS; Licciardi, F
2005 SEP ;84(11):S470-S471, Fertility & sterility
— id: 59580, year: 2005, vol: 84, page: S470, stat: Journal Article,

Low rates of ectopic pregnancy after in vitro fertilization (IVF): Do practice habits matter?
Keegan, DA; Flisser, E; Krey, LC; Noyes, N; Berkeley, AS; Grifo, JA
2005 SEP ;84(11):S130-S130, Fertility & sterility
— id: 59565, year: 2005, vol: 84, page: S130, stat: Journal Article,

Rearrangement of beta- and gamma- tubulin immunostaining in mouse MII oocytes in response to slow freezing and vitrification procedures
Liu, H; Greenseid, K; Grifo, J; Krey, L
2005 SEP ;84(11):S379-S379, Fertility & sterility
— id: 59575, year: 2005, vol: 84, page: S379, stat: Journal Article,

Choline replacement for sodium in vitrification and thaw solutions improves the viability and embryonic competence of mouse oocytes
Liu, H; Grifo, J; Krey, L
2005 SEP ;84(11):S36-S37, Fertility & sterility
— id: 59559, year: 2005, vol: 84, page: S36, stat: Journal Article,

A 10-year experience with preimplantation genetic diagnosis (PGD) at NYU fertility center
Talebian, S; Keegan, DA; Adler, A; Krey, LC; Grifo, JA
2005 SEP ;84(11):S333-S334, Fertility & sterility
— id: 59572, year: 2005, vol: 84, page: S333, stat: Journal Article,

DNA methylation patterns in human tripronucleate zygotes
Xu, Yanwen; Zhang, John J; Grifo, James A; Krey, Lewis C
2005 Mar;11(3):167-171, Molecular human reproduction
In mammals, the dynamic reprogramming of DNA methylation begins during gametogenesis and continues through embryogenesis. Recently, immunofluorescence staining with an antibody against 5-methylcytosine (anti-5-MeC) has revealed active demethylation of the male pronucleus in zygotes beginning at 4-6 h after fertilization. In this study, we characterized the DNA methylation patterns in mouse zygotes and in human tripronucleate (3 PN) zygotes discarded after conventional fertilization or following ICSI. Pronuclei were subjected to fluorescence in-situ hybridization to identify the X and/or Y chromosomes and then stained with anti-5-MeC. In diandric 3 PN zygotes from conventional IVF, we consistently observed one strongly and two weakly stained pronuclei. In contrast, the majority of 3 PN ICSI zygotes, mainly digynic zygotes, displayed two strongly and one weakly stained pronuclei. Two zygotes from ICSI failed to show any staining difference among the three pronuclei. Our results indicate that the active demethylation of male pronuclei occurs in both mouse and human zygotes. It is possible that the abnormal methylation patterns resulting from a dysfunctional cytoplasm may occur in a small number of oocytes and may affect embryonic viability
— id: 56045, year: 2005, vol: 11, page: 167, stat: Journal Article,

Gestational carrier pregnancy with oocytes obtained during surgery for stage IIIc ovarian cancer after controlled ovarian stimulation
Zhang, John; Grifo, James A; Del Priore, Giuseppe
2005 May;83(5):1547-1549, Fertility & sterility
OBJECTIVE: To report a case of gestational carrier pregnancy with oocytes from a stage IIIc ovarian cancer patient. DESIGN: Case report. SETTING: University hospital. PATIENT: A 38-year-old woman with stage IIIc ovarian cancer. INTERVENTION(S): Controlled ovarian stimulation, cancer surgery, and IVF-ET to a gestational carrier. MAIN OUTCOME MEASURE(S): Oocyte fertilization and pregnancy. RESULT(S): Singleton term delivery occurred after transfer of three frozen-thawed embryos. CONCLUSION(S): Cryopreservation of embryos derived from IVF of oocytes obtained from ovarian cancer patients should be an option for their future fertility
— id: 56044, year: 2005, vol: 83, page: 1547, stat: Journal Article,

Impact of cryopreservation and subsequent frozen embryo transfer (FET) on biopsied embryos for pre-implantation genetic diagnosis (PGD)
Adler, A; McCaffrey, C; Krey, L; Grifo, J
2004 SEP ;82(2):S244-S244, Fertility & sterility
— id: 48955, year: 2004, vol: 82, page: S244, stat: Journal Article,

Ooplasnnic factors are responsible for the developmental defects observed when immature cumulus-enclosed mouse oocytes are denuded prior to maturation
Chang, HC; Liu, H; Zhang, J; Grifo, J; Krey, LC
2004 SEP ;82(2):S10-S10, Fertility & sterility
— id: 48949, year: 2004, vol: 82, page: S10, stat: Journal Article,

Candidate lineage marker genes in human preimplantation embryos
Hansis, Christoph; Grifo, James A; Krey, Lewis C
2004 May;8(5):577-583, Reproductive biomedicine online
Cell allocation and subsequent lineage commitment in the human embryo may be established as early as in the unfertilized oocyte. This phenomenon might be the result of subtle differences of gene expression and protein distribution. To assess whether gene expression profiling by reverse transcription-polymerase chain reaction could be a suitable tool for the detection of cell allocation and lineage commitment, the expression pattern of the putative inner cell mass marker gene Oct-4 and the trophectodermal marker genes beta-HCG and beta-LH were correlated in individual blastomeres of preimplantation human embryos. In 2- to 5-cell stage embryos, expression of beta-HCG and Oct-4 mRNA was negatively correlated in all blastomeres with statistical significance, suggesting that cell allocation can be assessed by those markers at early stages. In 7- to 10-cell stage embryos, expression of beta-LH and Oct-4 mRNA was negatively correlated in some blastomeres without statistical significance, suggesting that more experiments are necessary to decide if lineage commitment can be assessed in some cells by those markers at later stages
— id: 47784, year: 2004, vol: 8, page: 577, stat: Journal Article,

Retinoblastoma in a child conceived by in vitro fertilisation
Lee, I; Finger, P T; Grifo, J A; Rausen, A R; Rebarber, A; Barad, D H
2004 Aug;88(8):1098-1099, British journal of ophthalmology
— id: 44951, year: 2004, vol: 88, page: 1098, stat: Journal Article,

Extending culture of frozen-thawed, cleavage stage embryos to blastocyst stage achieves higher pregnancy and implantation rates when compared to overnight culture
Ampeloquio, E; Adler, A; Lee, J; Krey, L; Grifo, J
2003 SEP ;80(11):S150-S150, Fertility & sterility
— id: 55400, year: 2003, vol: 80, page: S150, stat: Journal Article,

Influences of mouse strain and culture media on in vitro maturation and embryogenesis of denuded mouse oocytes
Chang, HC; Liu, H; Blaszczyk, A; Grifo, J; Krey, LC
2003 SEP ;80(11):S266-S266, Fertility & sterility
— id: 55404, year: 2003, vol: 80, page: S266, stat: Journal Article,

Day 28 serum beta-hCG levels and early pregnancy outcome
Keegan, DA; Krey, LC; Grifo, JA
2003 SEP ;80(11):S173-S173, Fertility & sterility
— id: 55401, year: 2003, vol: 80, page: S173, stat: Journal Article,

An oocyte donor's willingness to donate - does the recipient's lifestyle make a difference?
Kump, L; Licciardi, F; Krey, L; Noyes, N; Grifo, J; Berkeley, AS
2003 SEP ;80(11):S140-S140, Fertility & sterility
— id: 55399, year: 2003, vol: 80, page: S140, stat: Journal Article,

Germinal vesicle transfer rescues nuclear maturation from the cytoplasmic defects present in vitro-aged oocytes
Liu, H; Grifo, J; Krey, L
2003 SEP ;80(11):S258-S259, Fertility & sterility
— id: 55402, year: 2003, vol: 80, page: S258, stat: Journal Article,

Metaphase II nuclei generated by germinal vesicle transfer in mouse oocytes support embryonic development to term
Liu, Hui; Chang, Hung Chi; Zhang, John; Grifo, Jamie; Krey, Lewis C
2003 Sep;18(9):1903-1907, Human reproduction
BACKGROUND: Cytoplasmic defects are thought to cause aneuploidies in oocytes and embryos and oocyte 'reconstruction' by germinal vesicle (GV) transfer may circumvent such defects. In mice 'reconstructed' oocytes undergo meiosis and fertilize normally, but early embryonic development is compromised if their ooplasm matured in vitro. This study employs sequential MII spindle and/or pronucleus (PN) transfer to assess the embryonic potential of MII nuclei that form following GV transfer. METHODS AND RESULTS: Mouse embryos generated by these procedures were transferred to the oviducts of pseudopregnant mice to monitor pregnancy outcome. Following GV transfer, the resultant metaphase II (MII) nuclei were activated either in situ or transferred and activated in ooplasts from in-vivo matured oocytes. When exchanged with the female PN of a fertilized zygote, only the PNs that developed in in-vivo matured ooplasts generated live offspring. Viable offspring also resulted when MII nuclei were transferred to in-vivo matured ooplasts and fertilized by insemination with sperm or by artificial activation and male PN transfer. Significantly, the offspring displayed normal fertility as adults. CONCLUSION: This report of live births following GV transfer in mice illustrates the importance of the maturational history of the ooplasm at PN formation for normal embryonic and fetal development
— id: 38845, year: 2003, vol: 18, page: 1903, stat: Journal Article,

Offspring gender ratios differ between day 3 and day 5 embryo transfer
McCaffrey, C; Berkeley, A; Grifo, J; Kump, L; Licciardi, F; Noyes, N
2003 SEP ;80(11):S98-S98, Fertility & sterility
— id: 55397, year: 2003, vol: 80, page: S98, stat: Journal Article,

DNA methylation patterns in mouse and human zygotes
Xu, YW; Krey, L; Grifo, J; Zhang, J
2003 SEP ;80(11):S80-S80, Fertility & sterility
— id: 55396, year: 2003, vol: 80, page: S80, stat: Journal Article,

Pregnancy derived from human nuclear transfer
Zhang, J; Zhuang, GL; Zeng, Y; Acosta, C; Shu, YM; Grifo, J
2003 SEP ;80(11):S56-S56, Fertility & sterility
— id: 55395, year: 2003, vol: 80, page: S56, stat: Journal Article,

Dynamic methylation changes in the centromeric regions of chromosomes of mouse preimplantation embryos
Zhang, JJ; Xu, YW; Grifo, J; Krey, L
2003 SEP ;80(11):S264-S264, Fertility & sterility
— id: 55403, year: 2003, vol: 80, page: S264, stat: Journal Article,

Influences of gonadotropin stimulation on in vitro maturation and embryogenesis of denuded mouse oocytes
Chang, HC; Liu, H; Grifo, J; Krey, LC
2002 SEP ;78(3):S268-S268, Fertility & sterility
— id: 55579, year: 2002, vol: 78, page: S268, stat: Journal Article,

Assessment of beta-HCG, beta-LH mRNA and ploidy in individual human blastomeres
Hansis, Christoph; Grifo, James A; Tang, YaXu; Krey, Lewis C
2002 Sep-Oct;5(2):156-161, Reproductive biomedicine online
In human embryos, blastomeres differentiate into trophectoderm (TE) cells and inner cell mass (ICM) cells of blastocysts. Although morphologically indistinguishable, blastomeres at early cleavage stages are likely to undergo changes on a molecular level that make them destined to become ICM or TE cells. While the transcription factor Oct-4 might serve as a marker for totipotent ICM cells, human chorionic gonadotrophin might be used as the equivalent for TE cells. This study reports a reverse transcription-polymerase chain reaction procedure to assess human beta-HCG mRNA concentrations as well as ploidy in individual blastomeres from normally and abnormally fertilized human embryos. beta-HCG mRNA was detected in both euploid and aneuploid cells and in oocytes. Surprisingly, beta-LH mRNA was also detected in some euploid blastomeres. In regard to preimplantation genetic diagnosis, assessment of expression levels of beta-HCG and Oct-4 mRNA in individual biopsied cells might serve as a tool to identify embryogenic blastomeres in combination with testing for chromosome and single gene abnormalities
— id: 38846, year: 2002, vol: 5, page: 156, stat: Journal Article,

Germinal vesicle xeno-transfer between mouse and human oocytes: A model to study ooplasmic influences on meiotic division
Liu, H; Krey, LC; Zhang, J; Chang, HC; Grifo, J
2002 SEP ;78(3):S77-S77, Fertility & sterility
— id: 55567, year: 2002, vol: 78, page: S77, stat: Journal Article,

Blastocyst development on day 5 - A relationship to clinical pregnancy rate?
McCaffrey, C; Adler, A; Berkeley, AS; Grifo, J; Noyes, N; Krey, LC
2002 SEP ;78(3):S48-S48, Fertility & sterility
— id: 55565, year: 2002, vol: 78, page: S48, stat: Journal Article,

Germinal vesicle transfer between fresh and cryopreserved immature mouse oocytes
Moffa, Federica; Comoglio, Francesca; Krey, Lewis C; Grifo, James A; Revelli, Alberto; Massobrio, Marco; Zhang, John
2002 Jan;17(1):178-183, Human reproduction
BACKGROUND: We assessed the maturational competence and the chromosomal pattern of mouse oocytes reconstructed by germinal vesicle (GV) transfer technique using nuclear and/or cytoplasmic components from cryopreserved GV stage oocytes. METHODS: From 657 GV oocytes (326 fresh and 331 frozen/thawed), four groups of reconstructed oocytes were obtained by micromanipulation and electrofusion: fresh GV-fresh cytoplast (FF), thawed GV-thawed cytoplast (TT), fresh GV-thawed cytoplast (FT), thawed GV-fresh cytoplast (TF). All reconstructed oocytes were cultured in vitro to metaphase II. RESULTS: Survival rate after manipulation and electrofusion, as well as progression to metaphase II, did not differ significantly among the four groups. Comparing reconstructed oocytes with fresh and thawed control pools, the only difference was a slightly but significantly higher maturation rate in the TT pool versus matched controls (P < 0.01). Cytogenetic analysis of 25 reconstructed oocytes showed the expected number of 20 chromosomes in 88% of them. CONCLUSIONS: We conclude that both nuclear and cytoplasmic components derived from cryopreserved immature oocytes are suitable for GV transfer procedure, and generate chromosomally normal oocytes able to progress to metaphase II in vitro. The possibility of using cryostored immature oocytes as a source of nuclei and cytoplasm could help in applying GV transfer procedure, both in research and clinical settings
— id: 27281, year: 2002, vol: 17, page: 178, stat: Journal Article,

Major birth defects after assisted reproduction
Steinkampf, Michael P; Grifo, Jamie
2002 Oct 31;347(18):1449-1451, New England journal of medicine
— id: 38847, year: 2002, vol: 347, page: 1449, stat: Journal Article,

In vitro development of human triploid zygotes reconstructed by pronuclear transfer
Zhang, J; Shu, YM; Krey, LC; Liu, H; Zhuang, GL; Grifo, J
2002 SEP ;78(3):S180-S181, Fertility & sterility
— id: 55574, year: 2002, vol: 78, page: S180, stat: Journal Article,

The impact of severe oligospermia on blastocyst formation in IVF-ICSI
Adler, A; McCaffrey, C; Lu, L; Noyes, N; Grifo, J; Krey, L
2001 SEP ;76(3):S10-S10, Fertility & sterility
— id: 54913, year: 2001, vol: 76, page: S10, stat: Journal Article,

Cell cycle checkpoint proteins Bub1 and Mad2 localize to kinetochores during meiosis in mouse oocytes
Blaszczyk, A; Brockmann, C; Grifo, J; Krey, L
2001 SEP ;76(3):S260-S261, Fertility & sterility
— id: 54928, year: 2001, vol: 76, page: S260, stat: Journal Article,

Developmental potential and blastocyst formation rate in human embryos with early stage development delay, arrest, or with multinucleated blastomere (MNB)
Chi, L; DeJesus, E; McCaffery, C; Grifo, JA; Berkeley, AS; Krey, LC
2001 SEP ;76(3):S183-S183, Fertility & sterility
— id: 54925, year: 2001, vol: 76, page: S183, stat: Journal Article,

Uterine transplantation, abdominal trachelectomy, and other reproductive options for cancer patients
del Priore G; Smith JR; Boyle DC; Corless DJ; Zacharia FB; Noakes DA; Diflo T; Grifo JA; Zhang JJ
2001 Sep;943(3):287-295, Annals of the New York Academy of Sciences
More and more women with cancer issues are now raising fertility concerns as survival improves and childbearing is delayed. Pregnancy is no longer contraindicated in cancer patients including breast and endometrial cancer survivors. In fact, survival in patients treated for breast cancer who subsequently become pregnant is actually higher than that in patients who do not become pregnant. 'Therapeutic' abortions are no longer recommended. Assisted reproductive technology (ART) have been associated with ovarian neoplasms, but the association is probably not causal. Neither ART nor hormone replacement is contraindicated in cancer patients. Our institution is very supportive of patients and the difficult decisions cancer survivors face. Using a program of counseling and close collaboration between oncologists, perinatologists, and reproductive endocrinologists, informed patients are offered every possible option, including ART and uterine transplantation, to achieve their family planning objectives
— id: 26646, year: 2001, vol: 943, page: 287, stat: Journal Article,

Efficacy and safety of ganirelix acetate versus leuprolide acetate in women undergoing controlled ovarian hyperstimulation
Fluker, M; Grifo, J; Leader, A; Levy, M; Meldrum, D; Muasher, S J; Rinehart, J; Rosenwaks, Z; Scott, R T Jr; Schoolcraft, W; Shapiro, D B
2001 Jan;75(1):38-45, Fertility & sterility
OBJECTIVE: To assess the efficacy, safety, and local tolerance of ganirelix acetate for the inhibition of premature luteinizing hormone (LH) surges in women undergoing controlled ovarian hyperstimulation (COH). DESIGN: Phase III, multicenter, open-label randomized trial. SETTING: In vitro fertilization (IVF) centers in North America. PATIENT(S): Healthy female partners (n = 313) in subfertile couples for whom COH and IVF or intracytoplasmic sperm injection were indicated. INTERVENTION(S): Patients were randomized to receive one COH cycle with ganirelix or the reference treatment, a long protocol of leuprolide acetate in conjunction with follitropin-beta for injection. OUTCOME MEASURE(S): Number of oocytes retrieved, pregnancy rates, endocrine variables, and safety variables. RESULT(S): The mean number of oocytes retrieved per attempt was 11.6 in the ganirelix group and 14.1 in the leuprolide group. Fertilization rates were 62.4% and 61.9% in the ganirelix and leuprolide groups, respectively, and implantation rates were 21.1% and 26.1%. Clinical and ongoing pregnancy rates per attempt were 35.4% and 30.8% in the ganirelix group and 38.4% and 36.4% in the leuprolide acetate group. Fewer moderate and severe injection site reactions were reported with ganirelix (11.9% and 0.6%) than with leuprolide (24.4% and 1.1%). CONCLUSION(S): Ganirelix is effective, safe, and well tolerated. Compared with leuprolide acetate, ganirelix therapy has a shorter duration and fewer injections but produces a similar pregnancy rate
— id: 120798, year: 2001, vol: 75, page: 38, stat: Journal Article,

We are due for a correction...and we are working to achieve one
Grifo J; Hoffman D; McNamee PI
2001 Jan;75(1):14-14, Fertility & sterility
— id: 21254, year: 2001, vol: 75, page: 14, stat: Journal Article,

Analysis of Oct-4 expression and ploidy in individual human blastomeres
Hansis C; Tang YX; Grifo JA; Krey LC
2001 Feb;7(2):155-161, Molecular human reproduction
Oct-4, a decisive factor that maintains totipotency in murine embryonic and germ cells, is exclusively expressed in such cells. In mice, different levels of oct-4 expression in blastomeres predict development towards inner cell mass (ICM) (high oct-4) or trophectoderm (TE) (low oct-4). To address whether the mouse model also applies to human embryos, the cytoplasm of individual human blastomeres from normally and abnormally fertilized embryos was tested for Oct-4 expression by reverse transcription-polymerase chain reaction (RT-PCR). The nuclei of the same blastomeres were subjected to fluorescence in-situ hybridization (FISH) to determine ploidy. A significant difference in Oct-4 mRNA levels was revealed between blastomeres. The distribution of blastomeres with high Oct-4 levels varied according to the cleavage stage of the embryo: the more blastomeres, the lower the percentage with high Oct-4 levels. Aneuploid blastomeres did not exhibit lower Oct-4 mRNA levels than diploid ones. Thus, differential Oct-4 expression in individual human blastomeres appears to direct cells towards the ICM or TE lineages without regard to chromosomal status. Oct-4 might be used as a marker in preimplantation genetic diagnosis to identify embryogenic blastomeres
— id: 21255, year: 2001, vol: 7, page: 155, stat: Journal Article,

Tay-Sachs disease and preimplantation genetic diagnosis
Hansis, C; Grifo, J
2001 ;44:311-315, Advances in genetics
— id: 97656, year: 2001, vol: 44, page: 311, stat: Journal Article,

Fertility and maternal age strategies to improve pregnancy outcome
Krey L; Liu H; Zhang J; Grifo J
2001 Sep;943(3):26-33, Annals of the New York Academy of Sciences
In humans, the live birth rate drops precipitously with increasing maternal age, and this decline is associated with increases in the incidence of oocyte and embryo aneuploidy. Preimplantation aneuploidy screening has improved pregnancy outcome by significantly lowering the miscarriage rate. Nevertheless, aneuploidy screening only identifies the affected embryos; it does not attempt to correct the underlying biologic problem. Anomalies in chromosome segregation can result from a dysfunctional first or second meiotic division in the egg or develop after fertilization during the first few mitoses of early embryonic development. In both instances, ooplasmic anomalies may account for the nuclear problem. Low cell levels of cytoplasmic proteins (e.g., cytoskeletal elements, enzymes, energy stores, cell cycle regulatory proteins) may lead to a dysfunctional division of chromosomes during egg maturation or following fertilization. Ooplasmic injection is a micromanipulation technique that has produced pregnancies in patients with a history of poor-quality, fragmented embryos. Germinal vesicle transfer is a research procedure used to investigate the ooplasmic-nuclear interplay regulating cell cycle, maturation, and fertilization. Both these techniques may prove to be effective in improving the quality of eggs from patients of advanced maternal age
— id: 26647, year: 2001, vol: 943, page: 26, stat: Journal Article,

Poor embryo quality: The answer lies (mostly) in the egg
Krey, L C; Grifo, J A
2001 Mar;75(3):466-468, Fertility & sterility
— id: 120773, year: 2001, vol: 75, page: 466, stat: Journal Article,

A two- versus three-embryo transfer: the oocyte donation model
Licciardi F; Berkeley AS; Krey L; Grifo J; Noyes N
2001 Mar;75(3):510-513, Fertility & sterility
OBJECTIVE: To compare implantation and pregnancy rates in oocyte recipients undergoing a two-embryo versus three-embryo transfer, 3 days after retrieval. DESIGN: Retrospective comparative analysis. SETTING: University-based in vitro fertilization center. PATIENT(S): All oocyte recipients undergoing embryo transfer from January 1, 1997 through August 31, 1999. INTERVENTION(S): Recipients received two or three embryos. MAIN OUTCOME MEASURE(S): Implantation, and clinical and multiple pregnancy rates. RESULT(S): Seventy-three recipients underwent a two-embryo transfer, and 376 had three embryos replaced. The numbers of oocytes retrieved (12.7 +/- 0.89 vs. 13.1 +/- 0.36) and embryos obtained (8.05 +/- 0.65 vs. 8.77 +/- 0.27) did not differ between the two-embryo and three-embryo transfer groups, nor did the proportion of patients with embryo cryopreservation (54.3% vs. 42.6%, respectively). There was no significant difference in pregnancy or implantation rates when comparing those patients with a two-embryo transfer to those with a three-embryo transfer. Significantly, 13.8% of the pregnancies in the three-embryo transfer group were triplet. CONCLUSION(S): Reducing the number of embryos transferred in an oocyte donation cycle can lower the incidence of triplet pregnancies without significantly lowering the overall pregnancy rate
— id: 26775, year: 2001, vol: 75, page: 510, stat: Journal Article,

Ooplasmic influence on nuclear function during the metaphase II-interphase transition in mouse oocytes
Liu H; Krey LC; Zhang J; Grifo JA
2001 Dec;65(6):1794-1799, Biology of reproduction
Nuclear and pronuclear transfer procedures were used to assess the functional competence of the nucleus and cytoplasm of mouse germinal vesicle-stage oocytes denuded of granulosa cells and matured in vitro or in vivo before artificial activation using a sequential treatment of A23187 + cycloheximide. Following activation, in vitro-matured oocytes were 'fertilized' by inserting a male pronucleus (PN), cultured to the 2-cell stage, and then transferred to the oviducts of foster mothers. No live births were noted, whereas a 17% live birth rate was observed when in vivo-matured oocytes were used. The developmental competency of other zygotes was similarly assessed following the exchange of haploid PN of matured and activated eggs with the female PN of fertilized zygotes. When PN of oocytes subjected to maturation and activation in vitro were transferred, only 1 of 79 reconstructed zygotes developed to term. In contrast, the live birth rate was 21% (11 of 53) for zygotes reconstructed with PN from in vivo-matured oocytes. Moreover, a live birth rate of 23% (8 of 35) was observed for reconstructed zygotes with female PN from 'hybrid' oocytes created by transferring the metaphase II nuclei of in vitro-matured oocytes into enucleated, in vivo-matured oocytes before activation. Such results suggest that the nucleus of an in vitro-matured oocyte can support embryonic development, but only when it is activated in the proper ooplasmic milieu. The cellular factors creating this ooplasmic milieu appear to develop normally in vivo during follicle maturation to metaphase II, but they fail to do so when the oocytes are denuded of granulosa cells and cultured in vitro before the final stages of maturation. In parallel studies, male and female PN of in vivo-fertilized zygotes were inserted into oocytes that were activated and enucleated following either in vitro or in vivo maturation. Live birth rates were comparable at 19% (5 of 27) and 18% (9 of 49), respectively, suggesting that, regardless of the environment of the final stages of oocyte maturation, the resultant ooplasm is competent to support all aspects of embryonic development once activation and PN formation has been completed. Such findings only point further toward the importance of the condition of the ooplasmic milieu at the time of chemical activation. Whether a similar situation exists when eggs are activated following sperm penetration remains to be determined
— id: 26513, year: 2001, vol: 65, page: 1794, stat: Journal Article,

The nuclear developmental capacity of mouse oocytes following cryopreservation at germinal vesicle stage
Liu, H; Krey, LC; Zhang, J; Grifo, JA
2001 SEP ;76(3):S80-S80, Fertility & sterility
— id: 54917, year: 2001, vol: 76, page: S80, stat: Journal Article,

Factors useful in predicting the success of oocyte donation: a 3-year retrospective analysis
Noyes N; Hampton BS; Berkeley A; Licciardi F; Grifo J; Krey L
2001 Jul;76(1):92-97, Fertility & sterility
Objective: To establish prognostic relevance of parameters assessed in oocyte donation cycles.Design: Retrospective analysis.Setting: Large university-based donor oocyte program.Patient(s): All oocyte recipient cycles achieving embryo transfer from September 1995 to October 1998.Intervention(s): None.Main Outcome Measure(s): Pregnancy.Result(s): Recipient age and reproductive status, day 9 and 12 serum estradiol (E(2)) levels and a progesterone (P) level obtained 2 days after initiation of hormonal therapy did not correlate with pregnancy. Endometrial thickness, but not endometrial pattern, was useful in predicting pregnancy outcome. The clinical pregnancy and live-birth rate in cycles where the endometrial thickness was less than 8 mm was significantly lower when compared to cycles with an endometrial thickness >/=9 mm. Cycles where optimal quality embryos were transferred had the highest implantation (36%), clinical pregnancy (63%) and live birth (54%) rates and these rates were significantly higher than those of cycles where only poor quality embryos were available for transfer (10% implantation, 17% clinical pregnancy, and 8% live birth rates, respectively; P<.05).Conclusion(s): The most reliable predictive factors for pregnancy in oocyte donation cycles are the quality of the embryos transferred and the recipient's mid-cycle endometrial thickness. Recipient monitoring should minimally include ultrasound assessment of endometrial thickness
— id: 21151, year: 2001, vol: 76, page: 92, stat: Journal Article,

Preimplantation genetic diagnosis of cystic fibrosis (CF) with compound heterozgote S549R/DF508
Tang, Y; Krey, L; Adler, A; Chi, L; Grifo, J
2001 SEP ;76(3):S134-S134, Fertility & sterility
— id: 54920, year: 2001, vol: 76, page: S134, stat: Journal Article,

Preimplantation genetic diagnosis in two families at risk for recurrence of Herlitz junctional epidermolysis bullosa
Cserhalmi-Friedman, P B; Tang, Y; Adler, A; Krey, L; Grifo, J A; Christiano, A M
2000 Aug;9(4):290-297, Experimental dermatology
The Herlitz type of junctional epidermolysis bullosa (H-JEB) is a severe inherited bullous disease which leads to the early demise of the affected newborn. Mutations in the genes encoding the 3 polypeptides of the anchoring filament protein laminin 5 underlie this condition. We studied 2 families with affected children who previously died from H-JEB. Mutation screening using heteroduplex analysis and direct sequencing of the PCR products revealed a previously described hotspot mutation in LAMB3 (R635X), and a novel delayed termination codon in LAMB3 in the first proband. In the second proband, we found a novel initiation codon mutation in LAMB3, and a novel 2 bp deletion in LAMB3. For preimplantation genetic diagnosis (PGD) in these families, we developed nested multiplex PCR assays, amplifying the mutations and informative intragenic polymorphisms in the probands. Single embryonic cells were biopsied from 8-cell embryos using standard techniques, and subjected to the multiplex PCR assay followed by restriction enzyme digestion. Embryos found not to carry either mutation were transferred to the mothers, and a pregnancy was established in the second family as evidenced by the elevated level of HCG, although the pregnancy did not persist. This study illustrates the feasibility of PGD for an inherited skin disorder for the first time
— id: 120774, year: 2000, vol: 9, page: 290, stat: Journal Article,

Oct-4 expression in inner cell mass and trophectoderm of human blastocysts
Hansis C; Grifo JA; Krey LC
2000 Nov;6(11):999-1004, Molecular human reproduction
The expression of the transcription factor Oct-4 is thought to be one of the decisive factors that maintain totipotency in embryonic and germ cells. In mice, oct-4 is exclusively expressed in germ cells and totipotent cells of the embryo. In humans, Oct-4 is expressed in germ cells, embryonic stem cells and whole embryos at various stages of development. However, there is limited information about the distribution of Oct-4 expression in human embryos. In an attempt to address this issue, the inner cell mass (ICM) and trophectoderm (TE) of 17 human blastocysts were separated and Oct-4 mRNA expression individually assessed by reverse transcription-polymerase chain reaction (RT-PCR). In discarded blastocysts that developed from two pronuclear zygotes, the mean Oct-4 expression was 31 times higher in totipotent ICM cells than in differentiated TE cells. This finding suggests that, in accordance with data from the mouse, Oct-4 is highly expressed in human ICM cells as opposed to TE cells; this in turn supports the hypothesis that Oct-4 plays a similar role to maintain totipotency in these two species
— id: 47803, year: 2000, vol: 6, page: 999, stat: Journal Article,

Oct-4 expression in inner cell mass and trophectoderm of human blastocysts
Hansis, C; Tang, YX; McCaffrey, C; Grifo, J; Krey, L
2000 JUN ;15(8):91-91, Human reproduction
— id: 54495, year: 2000, vol: 15, page: 91, stat: Journal Article,

In-vitro development of mouse zygotes following reconstruction by sequential transfer of germinal vesicles and haploid pronuclei [In Process Citation]
Liu H; Zhang J; Krey LC; Grifo JA
2000 Sep;15(9):1997-2002, Human reproduction
We evaluated whether mouse oocytes reconstructed by germinal vesicle (GV) transfer can develop to blastocyst stage. The oocytes were artificially activated with sequential treatment of A23187 and anisomycin; fertilization was then established by transfer or exchange of pronuclei with those of zygotes fertilized in vivo. Type 1 zygotes were constructed by placing the male haploid pronucleus from a zygote into the cytoplasm of an oocyte that underwent GV transfer, in-vitro maturation and activation; for type 2 zygotes, the female pronucleus was removed from a zygote and replaced with the female pronucleus of an oocyte subjected to GV transfer, in-vitro maturation and activation. Karyotypes of activated oocytes and type 2 zygotes were also subjected to analysis. When cultured in human tubal fluid (HTF) medium, reconstructed oocytes matured and, following artificial activation, consistently developed a pronucleus with a haploid karyotype; the activation rate for this medium was two- to three-fold higher than that of oocytes cultured in M199 (87% versus 30% respectively). Following transfer of a male pronucleus, only 47% of the type 1 zygotes developed to morula or blastocyst stage and embryo morphology was poor. In contrast, 73% of the type 2 zygotes developed to morula or blastocyst stage, many even hatching, with few morphological anomalies. Normal karyotypes were observed in 88% of the type 2 zygotes analysed. These observations demonstrate that the nucleus of a mouse oocyte subjected to sequential nuclear transfer at GV and pronucleus stages is, nonetheless, capable of maturing meiotically, activating normally and supporting embryonic development to hatching blastocyst stage. In contrast, the developmental potential of the cytoplasm of such oocytes appears to be compromised by these procedures
— id: 11519, year: 2000, vol: 15, page: 1997, stat: Journal Article,

Successful preimplantation genetic diagnosis for compound heterozygote Tay-Sachs disease
Tang, YX; Hansis, C; Adler, A; Grifo, J
2000 JUN ;15(8):182-182, Human reproduction
— id: 54496, year: 2000, vol: 15, page: 182, stat: Journal Article,

Fertility after hysteroscopic resection of submucous myomas
Giatras K; Berkeley AS; Noyes N; Licciardi F; Lolis D; Grifo JA
1999 May;6(2):155-158, Journal of the American Association of Gynecologic Laparoscopists
STUDY OBJECTIVE: To analyze fertility outcomes after resection of submucous myomas by operative hysteroscopy in infertile women. DESIGN: Retrospective analysis (Canadian Task Force classification II-2). SETTING: Academic tertiary referral center. PATIENTS: Forty-one women (age 28-42 yrs) old with primary and secondary infertility, and histologically proved submucous myomas. Intervention. Hysteroscopic myomectomy performed with a rigid resectoscope. MEASUREMENTS AND MAIN RESULTS: Of the 41 patients, 25 (60.9%) became pregnant overall and 20 (48.7%) delivered at term. Seventeen patients delivered a single fetus. Five delivered twins, three at term and two at 33 and 35 weeks. One woman delivered triplets at 31 weeks. The total delivery rate was 56.0%. Two women miscarried, at 6 and 8 weeks. One patient developed postoperative Asherman's syndrome. CONCLUSION: Our results indicate that hysteroscopic myomectomy improves fertility in previously infertile women. Resection is a viable alternative to abdominal myomectomy for submucous myomas. (J Am Assoc Gynecol Laparosc 6(2):155-158, 1999)
— id: 56431, year: 1999, vol: 6, page: 155, stat: Journal Article,

Oral versus intramuscular progesterone for in vitro fertilization: a prospective randomized study
Licciardi FL; Kwiatkowski A; Noyes NL; Berkeley AS; Krey LL; Grifo JA
1999 Apr;71(4):614-618, Fertility & sterility
OBJECTIVE: To evaluate the efficacy of oral micronized progesterone compared with IM progesterone in oil for luteal support in patients undergoing IVF who are treated with a GnRH agonist. DESIGN: Randomized prospective clinical trial. SETTING: University-based IVF center. PATIENT(S): Women <40 years of age who were undergoing IVF with luteal GnRH pituitary down-regulation. INTERVENTION(S): Patients were randomized to receive either oral micronized progesterone (200 mg three times daily) or IM progesterone (50 mg daily). MAIN OUTCOME MEASURE(S): Progesterone levels at standardized days 21 and 28, and pregnancy and embryo implantation rates. RESULT(S): Day 21 progesterone levels were 77.6+/-13.2 ng/mL in the IM group and 81.5+/-16.2 ng/mL in the oral group. Day 28 progesterone levels were 76.3+/-15.0 ng/mL in the IM group and 53.6+/-10.1 ng/mL in the oral group. The clinical pregnancy rates were 57.9% and 45.8% for the IM and oral groups, respectively. The implantation rate per embryo was significantly higher in the IM group (40.9%) than in the oral group (18.1%). CONCLUSION(S): When used according to our protocols, oral progesterone and IM progesterone result in comparable levels of circulating progesterone. However, oral progesterone results in a reduced implantation rate per embryo
— id: 12025, year: 1999, vol: 71, page: 614, stat: Journal Article,

Reconstruction of mouse oocytes by germinal vesicle transfer: maturity of host oocyte cytoplasm determines meiosis
Liu H; Wang CW; Grifo JA; Krey LC; Zhang J
1999 Sep;14(9):2357-2361, Human reproduction
We evaluated the maturational competence of mouse oocytes reconstructed by the transfer and electrofusion of germinal vesicles (GV) into anuclear cytoplasts of GV stage oocytes (both auto- and hetero-transfers), metaphase II stage oocytes or zygotes. Following in-vitro culture, the maturation rates of the reconstructed oocytes to metaphase II did not significantly differ between auto- and hetero-transfers (40/70 versus 95/144 respectively); these rates also did not differ from those of control oocytes (57/97) which were matured in vitro without micromanipulation and electrofusion. In contrast, when a GV was transferred into an enucleated metaphase II oocyte or a zygote, only a few reconstructed oocytes underwent germinal vesicle breakdown (5/30 and 2/21 respectively); moreover, none reached metaphase II stage. Cytogenetic and immunofluorescence analyses were conducted on hetero-GV oocytes that extruded a first polar body. Each oocyte showed two groups of chromosomes, one in the cytoplast and one in the polar body, as well as a bipolar spindle with twenty univalent chromosomes. Our findings suggest that oocytes reconstructed by GV transfer into a cytoplast of the same developmental stage mature normally in vitro through metaphase II. Such oocytes may be a useful research model to elucidate the cytoplasmic and nuclear mechanisms regulating meiosis and the relationships between meiotic errors and age-related changes in the oocyte
— id: 6194, year: 1999, vol: 14, page: 2357, stat: Journal Article,

Elevated day 3 serum follicle stimulating hormone and/or estradiol may predict fetal aneuploidy
Nasseri A; Mukherjee T; Grifo JA; Noyes N; Krey L; Copperman AB
1999 Apr;71(4):715-718, Fertility & sterility
OBJECTIVE: To determine whether baseline serum FSH and/or E2 concentrations can predict the risk for fetal chromosomal abnormalities. DESIGN: Case control study. SETTING: Reproductive technology program at a university hospital. PATIENT(S): Patients who underwent dilation and curettage (D + C), and whose products of conception were karyotyped. INTERVENTION(S): Patients underwent natural conception or controlled ovarian hyperstimulation followed by intrauterine insemination, in vitro fertilization and embryo transfer, gamete intrafallopian transfer, or zygote intrafallopian transfer. MAIN OUTCOME MEASURE(S): Baseline serum FSH and E2 concentrations and fetal karyotype. RESULT(S): Genetic evaluation of 78 D + C specimens revealed 34 normal and 44 abnormal fetal karyotypes. A significantly greater proportion of women with abnormal fetal karyotype had elevated baseline serum FSH (> or =15 mIU/mL [RIA] or 10 mIU/mL [Immulite]) and/or E2 > or = 50 pg/mL [Immulite]) compared with women of normal fetal karyotype. Among karyotypically abnormal abortuses, autosomal trisomy was the most common abnormality noted (79.5%), followed by mosaicism (6.8%), triploidy (6.8%), monosomy XO (4.5%), and balanced translocation (2.3%). CONCLUSION(S): Baseline serum FSH and/or E2 concentrations may be valuable as predictors of fetal aneuploidy
— id: 22382, year: 1999, vol: 71, page: 715, stat: Journal Article,

In vitro fertilization outcome relative to embryo transfer difficulty: a novel approach to the forbidding cervix
Noyes N; Licciardi F; Grifo J; Krey L; Berkeley A
1999 Aug;72(2):261-265, Fertility & sterility
OBJECTIVE: To assess the impact of ET difficulty on IVF outcome and to optimize the ET procedure. DESIGN: Retrospective analysis of IVF outcome by ET catheter type and ET difficulty. Prospective treatment and follow-up of patients with a history of extremely difficult cervical passage. SETTING: Large university-based IVF program. PATIENT(S): All patients < 40 years of age undergoing IVF-ET from September 1995 to May 1998. INTERVENTION(S): Surgical correction of cervical stenosis. MAIN OUTCOME MEASURE(S): Pregnancy and embryo implantation rates. RESULT(S): Only 0.6% of ETs were 'extremely difficult.' Pregnancy rates were not statistically significantly different among ETs graded easy, moderate, and difficult. In contrast, no pregnancies occurred in the rare 'extremely difficult' ET group. Eight patients with a history of extremely difficult cervical passage underwent surgical correction of their cervical stenosis. Twelve postoperative IVF-ET in these women resulted in eight clinical pregnancies, six of which were multiple gestations. The embryo implantation rate of these cycles was 42.2%. CONCLUSION(S): Patients with a history of extremely difficult ET may benefit from hysteroscopic evaluation and possible modification of their cervical canal before a future IVF attempt
— id: 56464, year: 1999, vol: 72, page: 261, stat: Journal Article,

Electrical activation and in vitro development of human oocytes that fail to fertilize after intracytoplasmic sperm injection
Zhang J; Wang CW; Blaszcyzk A; Grifo JA; Ozil J; Haberman E; Adler A; Krey LC
1999 Sep;72(3):509-512, Fertility & sterility
OBJECTIVE: To determine whether electrically stimulated Ca2+ influx can 'rescue' fertilization and early embryogenesis in human oocytes that fail to fertilize after intracytoplasmic sperm injection (ICSI). DESIGN: Prospective, randomized trial of a laboratory procedure. SETTING: A research laboratory at a university medical center. PATIENT(S): Discarded oocytes from ICSI-IVF cycles. INTERVENTION(S): Oocytes (n = 104) that showed no evidence of fertilization 16-24 hours after ICSI were assigned to three treatment groups: group 1 (one direct current electrical pulse at 1.36-1.50 kV/cm for 40-60 micros), group 2 (three pulses every 15-20 minutes), or group 3 (treated the same as group 2 but with no electrical stimulation). MAIN OUTCOME MEASURE(S): After stimulation, the oocytes were cultured in vitro for 3-5 days. Oocytes that displayed two pronuclei and a second polar body within 16 hours were considered to have fertilized normally. Fertilization and embryo cleavage rates were compared between groups. RESULT(S): Fertilization occurred in 26 (70%) of 37 and 38 (78%) of 49 group 1 and 2 oocytes, respectively, but in only 5 (27%) of 18 group 3 oocytes. Within 3 days, group 2 embryos routinely developed beyond the two-cell to four-cell stage (61% versus 13% in group 1); 11% of these oocytes developed to the morula or early blastocyst stage. Sex chromosome analyses indicated 10 male and 8 female embryos. CONCLUSION(S): Oocytes that fail to fertilize by 24 hours after ICSI can resume apparently normal fertilization and early embryonic development in response to electrical stimulation. Moreover, the degree of cytoplasmic activation as determined by the number of pulses applied affects fertilization efficiency and early embryonic development
— id: 11947, year: 1999, vol: 72, page: 509, stat: Journal Article,

In vitro maturation of human preovulatory oocytes reconstructed by germinal vesicle transfer
Zhang J; Wang CW; Krey L; Liu H; Meng L; Blaszczyk A; Adler A; Grifo J
1999 Apr;71(4):726-731, Fertility & sterility
OBJECTIVE: To describe a micromanipulation-electrofusion procedure for transferring germinal vesicles (GVs) between immature human oocytes. DESIGN: Pilot study to assess oocyte maturation after an invasive micromanipulation procedure. SETTING: Research laboratory at a university medical center. PATIENT(S): Immature oocytes were discarded from intracytoplasmic sperm injection (ICSI)-IVF cycles of patients 23-48 years of age. INTERVENTION(S): Initially, GV removal and transfer were performed on the same oocyte; these 'self-reconstructed' oocytes were then cultured in vitro for up to 50 hours and examined periodically for maturation as judged by the extrusion of the first polar body. In a second study, GVs from oocytes of 'old' patients (>38 years old) were successfully transferred into enucleated immature oocytes of 'young' patients (<31 years old). MAIN OUTCOME MEASURE(S): Extrusion of the first polar body was monitored in 'reconstructed' and control oocytes; karyotypes also were analyzed at meiosis II. RESULT(S): From 48 oocytes from old patients, 12 GVs were successfully removed, transferred, and fused into previously enucleated oocytes from young patients. After in vitro culture, 7 of these 'reconstructed' oocytes matured to meiosis II, a maturation rate not significantly different from that observed in nonmanipulated controls. A normal, second meiotic metaphase chromosome complement was observed in 4 of 5 reconstructed oocytes. CONCLUSION(S): Normal meiosis can occur after the transfer of a GV into an enucleated host oocyte. Germinal vesicle transfer may be a valuable research procedure that generates cell models to characterize the cytoplasmic-nuclear interplay for cell cycle regulation, maturation, and fertilization in the human oocyte; it also may be a potentially attractive alternative to oocyte donation
— id: 56418, year: 1999, vol: 71, page: 726, stat: Journal Article,

Simultaneous assessment of sperm chromatin condensation and morphology before and after separation procedures: effect on the clinical outcome after in vitro fertilization
Angelopoulos T; Moshel YA; Lu L; Macanas E; Grifo JA; Krey LC
1998 Apr;69(4):740-747, Fertility & sterility
OBJECTIVE: To look for correlations between acridine orange (AO) staining and semen parameters before and after sperm separation procedures and to assess whether the AO test predicts fertilization or pregnancy outcomes after standard IVF and intracytoplasmic sperm injection. DESIGN: Prospective study that simultaneously assesses sperm morphology and nuclear protein maturity on a cell-by-cell basis before and after preparative procedures. SETTING: University teaching hospital. PATIENT(S): Men (n = 140) undergoing diagnostic semen analysis. MAIN OUTCOME MEASURE(S): Acridine orange fluorescence of sperm nuclei, semen parameters, IVF outcome. RESULT(S): In unprocessed samples, 90% of sperm with normal heads displayed green fluorescence (mature nuclear protein); significantly lower percentages of green fluorescence were observed in sperm with abnormal heads. The percentage of mature normal sperm in the specimen correlated with motility. Sperm maturity after swim-up or Percoll gradient was significantly improved for sperm with normal or abnormal heads. The percentage of mature normal sperm correlated with motility after either Percoll or swim-up. Neither the percentages of mature nuclei nor mature normal nuclei correlated with fertilization or pregnancy outcome. CONCLUSION(S): Nuclear protein maturation correlates with sperm motility and morphology. Because morphologically normal and motile sperm are more mature, separation procedures should generate a population of sperm with the highest fertilization capacity. Acridine orange staining, however, did not predict fertilization efficiency or pregnancy outcome in IVF cycles
— id: 57094, year: 1998, vol: 69, page: 740, stat: Journal Article,

Preimplantation genetic diagnosis of human embryos for Marfan's syndrome
Blaszczyk A; Tang YX; Dietz HC; Adler A; Berkeley AS; Krey LC; Grifo JA
1998 May;15(5):281-284, Journal of assisted reproduction & genetics
PURPOSE: Single-cell nested polymerase chain reaction (PCR) and Ddel endonuclease digestion were used to detect the presence of a Marfan's syndrome mutation in human preimplantation embryos derived from in vitro fertilization (IVF). These procedures were conducted to eliminate the possibility of transmission of the affected allele from the father to his offspring. The mutation on chromosome 15 is transmitted as an autosomal dominant trait, and the chance of having a child affected with the disease is 50%. METHODS: A couple presented to the Program for In Vitro Fertilization, Reproductive Surgery and Infertility for preimplantation genetic diagnosis. IVF was performed and embryo biopsy was done on day 3 embryos. Single blastomeres were removed from embryos and subjected to nested PCR analysis and endonuclease digestion to detect a Marfan's syndrome mutation located on chromosome 15 inherited from the father. RESULTS: Thirteen oocytes were injected with spermatozoa using intracytoplasmic sperm injection, and nine fertilized normally. Following embryo biopsy and polymerase chain reaction amplification-Ddel endonuclease digestion, five embryos were detected that were positive for the mutation. The four non-affected embryos were transferred to the uterus, resulting in a healthy and normal ongoing pregnancy
— id: 7505, year: 1998, vol: 15, page: 281, stat: Journal Article,

Preimplantation genetic diagnosis in a family at risk for recurrence of Herlitz junctional epidermolysis bullosa
Cserhalmi-Friedman, PB; Tang, YX; Grifo, JA; Christiano, AM
1998 APR ;110(4):485-485, Journal of investigative dermatology
— id: 98350, year: 1998, vol: 110, page: 485, stat: Journal Article,

Laparoscopy for pelvic pain in the Mayer-Rokitansky-Kuster-Hauser syndrome. A case report
Giatras K; Licciardi F; Grifo JA
1998 Mar;43(3):203-205, Journal of reproductive medicine
BACKGROUND: The Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome occurs in 1 of every 4,000-5,000 female births. It is characterized by normal external genitalia, an absent vagina, absent or rudimentary uterus, and normal fallopian tubes and ovaries. When associated with a rudimentary uterine horn, cyclic catamenial pelvic pain may result. The standard procedure for pain relief has been removal of the uterine horn by laparotomy. CASE: A rudimentary uterine horn was diagnosed in a woman with MRKH syndrome who developed monthly severe pelvic pain. Removal of the structure was performed via laparoscopy. The patient had complete resolution of her pain. CONCLUSION: As an alternative to laparotomy, laparoscopic resection of a rudimentary horn in patients with MRKH syndrome is both feasible and beneficial in the treatment of pelvic pain
— id: 57154, year: 1998, vol: 43, page: 203, stat: Journal Article,

Successful outcome with day 4 embryo transfer after preimplantation diagnosis for genetically transmitted diseases
Grifo JA; Giatras K; Tang YX; Krey LC
1998 Jun;13(6):1656-1659, Human reproduction
Preimplantation genetic diagnosis was performed in 61 day 3 embryos obtained by in-vitro fertilization from seven patient carriers of haemophilia, Marfan's syndrome, Bloch-Sulzemberg syndrome (incontinentia pigmentosa) or X chromosome-linked immune deficiency, retinitis pigmentosa, and FG syndrome, which is characterized by mental retardation and hypotonia. After multiplex polymerase chain reaction, 16 embryos were diagnosed as being unaffected, and these were transferred to the uterus on the following day (day 4). Of these embryos, six (37.5%) implanted, resulting in the delivery of a singleton and a twin pregnancy, a late second trimester miscarriage (twins at week 20) and a first trimester miscarriage at week 8. All the diagnoses were confirmed by amniocentesis. We report for the first time a late day 4 transfer of biopsied human embryos undergoing preimplantation genetic diagnosis. This transfer schedule allows an extra day to perform genetic analyses on single blastomeres and to monitor any adverse effect of the biopsy procedure
— id: 7584, year: 1998, vol: 13, page: 1656, stat: Journal Article,

Genetics, age, and infertility
Nasseri A; Grifo JA
1998 Oct 12;30(2):189-192, Maturitas
OBJECTIVE: To review the genetics of aging specifically as it pertains to human fertility, as well as the recent advancements in the diagnosis of genetic diseases prior to embryo implantation. METHODS: A review of our own experience as well as the scientific literature with regards to the decline in female fertility with age, the success of IVF in women of older reproductive age, and the role of preimplantation genetic diagnosis (PGD) in the evaluation of the patient at risk for fetal genetic anomalies. RESULTS: The decline in female fertility occurs primarily as a result of a decline in oocyte quality as well as quantity. The frequency of chromosomal anomalies in recognized abortuses increases in parallel with the age-specific rise in the incidence of spontaneous abortions. PGD is an accurate diagnostic tool for exclusion of genetically deficient embryos prior to initiation of pregnancy. CONCLUSION: Reproductive failure in women of older age appears to be directly related to ovarian age. Recent techniques such as cytoplasmic or germinal vesicle transfer are designed to replace the senescent cellular machinery believed to be responsible for genetic errors that occur during early cell division. PGD can accurately identify embryos with genetic deficiencies prior to implantation
— id: 22383, year: 1998, vol: 30, page: 189, stat: Journal Article,

Genetic screening of prospective oocyte donors
Wallerstein R; Jansen V; Grifo JA; Berkeley AS; Noyes N; Licker J; Licciardi F
1998 Jul;70(1):52-55, Fertility & sterility
OBJECTIVE: To report our experience with genetic screening of oocyte donor candidates and to determine the frequency with which significant genetic issues are identified. DESIGN: Prospective genetic screening of oocyte donor candidates. SETTING: University hospital oocyte donation program. PATIENT(S): Women presenting consecutively as volunteer oocyte donors. INTERVENTION(S): Genetic screening was performed by pedigree analysis and laboratory studies. MAIN OUTCOME MEASURE(S): Inclusion in the oocyte donor pool based on the results of clinical evaluation and laboratory tests consisting of polymerase chain reaction based mutational analysis for cystic fibrosis carrier status, cytogenetic analysis for karyotype, enzymatic assay for Tay-Sachs disease carrier status, and complete blood count and hemoglobin electrophoresis. RESULT(S): Eight (11%) of 73 oocyte donor candidates were excluded from the donor pool because of a potentially serious genetic finding. Cystic fibrosis mutations were identified in 5 candidates (7%), abnormal karyotypes were found in 2 (3.5%), and an autosomal dominant skeletal dysplasia was identified in 1 (1.4%). CONCLUSION(S): A significant proportion of women who present as candidates for oocyte donation are inappropriate for donation because of their genetic history or genetic testing results. A thorough genetic evaluation, including a history and laboratory screening, is essential to any oocyte donation program to maximize positive outcomes in pregnancies achieved through assisted means
— id: 12098, year: 1998, vol: 70, page: 52, stat: Journal Article,

A simple and objective approach to identifying human round spermatids
Angelopoulos T; Krey L; McCullough A; Adler A; Grifo JA
1997 Oct;12(10):2208-2216, Human reproduction
Although round spermatids have been studied extensively using staining techniques and electron microscopy, little information is available about their appearance in living conditions. We describe a method of collecting and identifying round spermatids from ejaculates and testicular biopsies. The validity of the selection procedure was confirmed by fluorescence in-situ hybridization. Based on cell size, morphological characteristics of nucleus and cytoplasm, and on the nucleus/cytoplasm ratio, we harvested a population of cells that was 84% haploid. This procedure can be applied to select spermatids for clinical or research purposes
— id: 7484, year: 1997, vol: 12, page: 2208, stat: Journal Article,

A morphological and cytogenetic study of the germinal cells in male cancer patients
Angelopoulos, T; Krey, L; McCullough, A; Adler, A; Grifo, JA
1997 JUN ;12(2-3):P28-P28, Human reproduction
— id: 53176, year: 1997, vol: 12, page: P28, stat: Journal Article,

A simple and objective approach to identifying human round spermatids
Angelopoulos, T; Krey, L; McCullough, A; Adler, A; Grifo, JA
1997 JUN ;12(2-3):P27-P27, Human reproduction
— id: 53175, year: 1997, vol: 12, page: P27, stat: Journal Article,

Correlation between semen parameters and maturity of normal human spermatozoa as assessed by Acridine Orange staining
Angelopoulos, T; Moshel, YA; Lu, L; Torres, L; Krey, LC; Grifo, JA
1997 JUN ;12(2-3):P70-P70, Human reproduction
— id: 53177, year: 1997, vol: 12, page: P70, stat: Journal Article,

Laparoscopic resection of a noncommunicating rudimentary uterine horn
Giatras K; Licciardi FL; Grifo JA
1997 Aug;4(4):491-493, Journal of the American Association of Gynecologic Laparoscopists
Two women had infertility due to a symptomatic unicornuate uterus associated with rudimentary contralateral horn. Both carried successful pregnancies after laparoscopic resection of the horns
— id: 56949, year: 1997, vol: 4, page: 491, stat: Journal Article,

Update in preimplantation genetic diagnosis. Age, genetics, and infertility
Grifo JA; Tang YX; Krey L
1997 Sep 26;828:162-165, Annals of the New York Academy of Sciences
PGD has been successfully used for several years. Over 40 babies have been born worldwide by use of these techniques. Unfortunately, a number of misdiagnoses have been made, a distressing consequence of a new frontier. Significant advances have been made to improve the efficiency and accuracy of PCR and FISH. The widespread use of this technology awaits further documentation of safety and accuracy. Other issues must also be addressed. First, the cost-effectiveness of the techniques relative to the traditional alternatives must be evaluated. A number of ethical issues regarding embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for nongenetic disorders such as gender selection. Finally, the experimental nature of these procedures must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved IVF success rates may make PGD a more widely used clinical tool. The future awaits these developments
— id: 12257, year: 1997, vol: 828, page: 162, stat: Journal Article,

Update in preimplantation genetic diagnosis: successes, advances, and problems
Grifo JA; Tang YX; Munne S; Krey L
1996 Apr;8(2):135-138, Current opinion in obstetrics & gynecology
The field of preimplantation genetic diagnosis has undergone significant advances since the report of the first birth from this method in 1990. The first birth in the USA was reported in 1992, as was the first successful diagnosis and delivery of a baby free of a single gene defect disorder (cystic fibrosis and then Tay Sachs). Investigators have now reported approximately 40 births worldwide from preimplantation genetic diagnosis using the polymerase chain reaction and fluorescent in-situ hybridization methods to analyze single cells removed from early cleavage stage preimplantation embryos. The International Working Group on Preimplantation Genetics meets annually to discuss progress and pitfalls in this field. Although preimplantation genetic diagnosis offers hope to patients at risk of transmitting disease, there are many technical hazards of this experimental procedure. Technical difficulties must be overcome in order for preimplantation genetic diagnosis to become a standard clinical tool. This review will highlight some of the recent advances and problems in the field of preimplantation genetic diagnosis
— id: 12628, year: 1996, vol: 8, page: 135, stat: Journal Article,

Expression of the 60 kDa heat shock protein in peritoneal fluids from women with endometriosis: implications for endometriosis-associated infertility
Kligman, I; Grifo, J A; Witkin, S S
1996 Dec;11(12):2736-2738, Human reproduction
Proinflammatory cytokines and activated macrophages and T lymphocytes have been detected in peritoneal fluids of women with endometriosis and may impair fertility. Expression of the 60 kDa heat shock protein (hsp60) is one mechanism leading to a localized activation of macrophages and T lymphocytes and cytokine release. Peritoneal fluids, obtained from 68 women undergoing a diagnostic laparoscopy, were assayed for hsp60. As independent evidence of local immune activation, the fluids were analysed for interferon gamma (IFN gamma). Fluids were also tested for antibodies to Chlamydia trachomatis because a chronic asymptomatic infection by this organism may also release hsp60. At laparoscopy, 26 women were diagnosed with pelvic adhesions, 19 had endometriosis, 16 had a visibly normal pelvis, four had ovarian cysts while three had myomas. The prevalence of hsp60 was higher in peritoneal fluids from the women with endometriosis than in the other subjects (P = 0.005). Hsp60 was detected in seven (36.8%) of the endometriosis patients and in only one each of the women with adhesions, a normal pelvis or an ovarian cyst; all women with myomas were negative. Detection of IFN gamma in peritoneal fluids was highly correlated with the presence of hsp60 (P = 0.0003). IFN gamma was present in seven of nine (77.8%) women with hsp60 and in only five of 40 (12.5%) women lacking hsp60. Women with pelvic adhesions had an increased prevalence of immunoglobulin G antibodies to C.trachomatis compared with the other women (P = 0.01). There was no relationship between evidence of exposure to C.trachomatis and hsp60 in peritoneal fluids. These data suggest that hsp60 may be released into the peritoneal fluid as a consequence of implanted ectopic endometrium. Hsp60-mediated immune activation may be one mechanism leading to endometriosis-associated infertility
— id: 120775, year: 1996, vol: 11, page: 2736, stat: Journal Article,

Marfan syndrome as a paradigm for transcript-targeted preimplantation diagnosis of heterozygous mutations
Eldadah, Z A; Grifo, J A; Dietz, H C
1995 Aug;1(8):798-803, Nature medicine
Among the many clinical applications of the polymerase chain reaction (PCR) is its potential use in preimplantation diagnosis of genetic disorders. Performing PCR on single blastomeres from early cleavage stage (six- to eight-cell) human embryos should, in principle, enable reliable determination of disease status for certain inherited conditions. However, reports of misdiagnoses using this technique have diminished enthusiasm for its widespread clinical use. One principal source of error is the propensity for genome-targeted PCR to exclusively amplify one allele in reactions assaying a single heterozygous diploid cell. Complete reaction failure is also common. Employing the Marfan syndrome (MFS) as a paradigm, we have developed a reliable, reverse transcription-PCR-based method of genotyping single cells that overcomes these obstacles. The technique should facilitate accurate preimplantation diagnosis of MFS and other selected genetic diseases caused by heterozygous or compound-heterozygous mutations
— id: 120777, year: 1995, vol: 1, page: 798, stat: Journal Article,

Laser ablation of the mouse zona pellucida for blastomere biopsy
Licciardi F; Gonzalez A; Tang YX; Grifo J; Cohen J; Neev Y
1995 Aug;12(7):462-466, Journal of assisted reproduction & genetics
— id: 66619, year: 1995, vol: 12, page: 462, stat: Journal Article,

The use of first polar bodies for preimplantation diagnosis of aneuploidy
Munne S; Dailey T; Sultan KM; Grifo J; Cohen J
1995 Apr;10(4):1014-1020, Human reproduction
A large proportion of patients undergoing in-vitro fertilization (IVF) are aged > or = 35 years. It has been estimated that in this age group, 50% of embryos are chromosomally abnormal, with aneuploidy being the major contributing factor. Since the origin of most aneuploidies is maternal meiosis I non-disjunction, unfertilized oocytes could be safely screened for aneuploidy by analysing their first polar bodies. To determine the feasibility of first polar body aneuploidy analysis, polar bodies were analyzed by fluorescence in-situ hybridization (FISH) using probes simultaneously for chromosomes X, Y, 18, 13/21 or X, Y, 18 and 16. Within 6 h of retrieval, 88% showed a normal segregation involving a single chromosome of each kind, with double-dotted hybridization signals, corresponding to dyads (chromosomes in metaphase I composed of two chromatids). The rest showed non-disjunction of full dyads (6%), or an unbalanced pre-division of dyads (6%), which gives a segregation of one chromatid or one dyad and a chromatid with the first polar body. But only 34% of polar bodies analysed 24 h after retrieval or later showed a normal segregation, with most of the other polar bodies showing balanced pre-division, with two separated hybridization signals for all the chromosomes analysed. The rates of non-disjunction and unbalanced pre-division after > or = 24 h in culture were similar to the rates in fresh oocytes. When both types of aneuploidy were considered together, an increase of aneuploidy with maternal age was detected, which although slight, was significant (P = 0.025).(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 20780, year: 1995, vol: 10, page: 1014, stat: Journal Article,

Assessment of numeric abnormalities of X, Y, 18, and 16 chromosomes in preimplantation human embryos before transfer
Munne S; Sultan KM; Weier HU; Grifo JA; Cohen J; Rosenwaks Z
1995 Apr;172(4 Pt 1):1191-1199, American journal of obstetrics & gynecology
OBJECTIVE: Our purpose was to determine the feasibility of ascertaining aneuploidy for chromosomes X, Y, 18, and 16 by use of multiple-probe fluorescence in situ hybridization in blastomeres from preimplantation human embryos. STUDY DESIGN: A short fluorescence in situ hybridization procedure involving the simultaneous use of four deoxyribonucleic acid probes detected with red, green, blue, or a mixture of red and green fluorochromes was developed to determine numeric abnormalities of chromosomes X, Y, 18, and 16. Embryos underwent biopsy, and all or most cells were analyzed to distinguish true aneuploidy from mosaicism and to assess technique variations within the same embryo (n = 64). RESULTS: The analysis of all the blastomeres of an embryo was achieved in 91% of the embryos. Successful analyses including biopsy, fixation, and fluorescence in situ hybridization were achieved in 87.8% of the blastomeres. Of the four chromosomes tested, numeric aberrations were found in 23% and 42% of normally and abnormally developing embryos, respectively, including aneuploidy, polyploidy, haploidy, and mosaicism. When diploid embryos containing one or several tetraploid cells are counted as chromosomally abnormal, then 49% and 61% of normally and abnormally developing embryos, respectively, were chromosomally abnormal. Aneuploid embryos consisted of two monosomies for chromosome 16, one for chromosome 18, and a trisomy for chromosome 16. There was a tendency for aneuploidy to increase with maternal age. CONCLUSIONS: Fluorescence in situ hybridization is a more efficient method than cytogenetic analysis to study specific aneuploidies at preimplantation stages of development in human embryos. In addition, the preimplantation genetic diagnosis of two blastomeres per eight-cell embryo may be sufficient to ensure successful analysis of polyploidy, haploidy, and specific aneuploidies without endangering the survival of the embryo. The technique can be easily modified to consider other chromosomes, including 13 and 21. Because most chromosomally abnormal embryos do not develop to term, the use of this technique may increase the delivery rate per embryo by allowing only transfer of embryos normal for the tested chromosomes. This technique would be most useful for older women undergoing in vitro fertilization, because aneuploidy appears to increase with advancing maternal age
— id: 20779, year: 1995, vol: 172, page: 1191, stat: Journal Article,

Laparoscopic resection of a noncommunicating rudimentary uterine horn. A case report
Schattman, G L; Grifo, J A; Birnbaum, S
1995 Mar;40(3):219-220, Journal of reproductive medicine
Patients who have a unicornuate uterus with a noncommunicating rudimentary horn that contains an endometrial cavity are at risk for endometriosis and obstetric complications. As in this case, resection of the rudimentary horn can be performed laparoscopically without increased risk to the patient and with some potential benefit
— id: 120779, year: 1995, vol: 40, page: 219, stat: Journal Article,

Chlamydia trachomatis detected by polymerase chain reaction in cervices of culture-negative women correlates with adverse in vitro fertilization outcome
Witkin, S S; Kligman, I; Grifo, J A; Rosenwaks, Z
1995 Jun;171(6):1657-1659, Journal of infectious diseases
The prevalence of Chlamydia trachomatis in the endocervices of 307 asymptomatic culture-negative women undergoing in vitro fertilization (IVF) was evaluated. C. trachomatis was detected by polymerase chain reaction (PCR) in 20 subjects (6.5%), and there were strong correlations between a positive finding and both failure to become pregnant (P = .013) and spontaneous abortion after embryo transfer (P = .004). C. trachomatis was identified in 2 (1.8%) of 112 who had term deliveries, 3 (27.3%) of 11 who spontaneously aborted, 1 (3.3%) of 30 with biochemical pregnancies, 13 (9.6%) of 135 with no pregnancy after embryo transfer, and 1 (5.3%) of 19 whose embryos did not become fertilized. There were no relationships between PCR findings and maternal age, cause of infertility, number of oocytes retrieved or fertilized, or number of embryos transferred; 55% of PCR-positive and 40% of PCR-negative women were undergoing at least their second IVF. An undetected C. trachomatis infection may be responsible for implantation failure or spontaneous abortion after IVF and embryo transfer
— id: 120778, year: 1995, vol: 171, page: 1657, stat: Journal Article,

Ureaplasma urealyticum and Mycoplasma hominis detected by the polymerase chain reaction in the cervices of women undergoing in vitro fertilization: prevalence and consequences
Witkin, S S; Kligman, I; Grifo, J A; Rosenwaks, Z
1995 Oct;12(9):610-614, Journal of assisted reproduction & genetics
PURPOSE: The prevalence of Ureaplasma urealyticum and Mycoplasma hominis in the endocervix at the time of oocyte collection in women undergoing in vitro fertilization (IVF) was examined using the polymerase chain reaction (PCR). METHODS: All women were treated with tetracycline following sample collection. RESULTS: U. urealyticum was identified in 56 (17.2%) of 326 women while M. hominis was present in only 5 (2.1%) of 235 women. U. urealyticum was detected at a higher frequency (P = 0.01) in those women whose IVF cycle failed prior to embryo transfer. This organism was present in 8 of 19 (42.1%) women with either no fertilization or no embryo transfer, 19 of 148 (12.8%) who had no evidence of pregnancy following embryo transfer, 6 of 30 (20.0%) who had only a transient (biochemical) pregnancy, 5 of 14 (35.7%) with a spontaneous abortion, and 18 of 115 (15.6%) with a term birth. Of the eight women with U. urealyticum who had no embryos transferred, male factor was the cause of infertility in five cases, two women had tubal occlusions while in one woman the diagnosis was idiopathic. Therefore, poor sperm quality, and not a U. urealyticum infection, might explain the failure of most of these cases to proceed to the stage of embryo transfer. Analysis of all patients revealed no association between male factor infertility and U. urealyticum in the cervix. CONCLUSIONS: U. urealyticum, but not M. hominis, is present in the cervices of many culture-negative women. Its presence, however, does not influence IVF outcome subsequent to embryo transfer in women treated with tetracycline after oocyte retrieval
— id: 120776, year: 1995, vol: 12, page: 610, stat: Journal Article,

Healthy deliveries from biopsied human embryos
Grifo JA; Tang YX; Munne S; Alikani M; Cohen J; Rosenwaks Z
1994 May;9(5):912-916, Human reproduction
Preimplantation genetic diagnosis was performed in 122 embryos obtained by IVF from 11 patients carriers of haemophilia, Duchenne's muscular dystrophy, Barth's syndrome, cystic fibrosis, Pelizaeus-Merzbacher syndrome or Rett's syndrome. After multiplex polymerase chain reaction (PCR) or fluorescence in situ hybridization (FISH) analysis with multiple probes, 28 embryos diagnosed as not affected were replaced. Of these, eight implanted (28%) and produced three ongoing pregnancies, three deliveries of four babies and a biochemical pregnancy. However, one case screened for cystic fibrosis was misdiagnosed and the pregnancy was terminated. In order to evaluate the efficiency of multiplex PCR, 55 non-replaced embryos were reassessed by PCR or by FISH. Identical results were obtained in all cases. However, one embryo which had only X-chromosome specific amplification by PCR was found to be XO in all its cells by FISH. Although multiplex PCR is demonstrated to be reliable for sexing of human embryos, FISH has the additional advantages of supplying ploidy assessment while not being affected by contamination
— id: 66620, year: 1994, vol: 9, page: 912, stat: Journal Article,

Sex determination of human embryos using the polymerase chain reaction and confirmation by fluorescence in situ hybridization
Munne S; Tang YX; Grifo J; Rosenwaks Z; Cohen J
1994 Jan;61(1):111-117, Fertility & sterility
OBJECTIVE: To use fluorescence in situ hybridization to corroborate the polymerase chain reaction (PCR) preimplantation diagnosis of human embryos in three couples carrying a chromosome X-linked disease. SETTING: Clinical and research IVF laboratories. PATIENTS: Individuals undergoing preimplantation diagnosis. RESULTS: Four ETs were performed in couples undergoing preimplantation diagnosis by multiplex PCR or fluorescence in situ hybridization, resulting in the birth of two normal female twins. The result of another is pending. A total of 22 embryos were analyzed by PCR. Embryos that were diagnosed as being at risk of carrying the genetic abnormality (n = 8), embryos that failed diagnosis (n = 4), and genetically normal embryos that arrested development (n = 4) were further analyzed by fluorescence in situ hybridization. The sex of all 16 embryos was determined and confirmed the previous 12 preimplantation diagnoses by multiplex PCR. In addition, fluorescence in situ hybridization analysis allowed the detection of two aneuploid embryos, one XO and one XXY, previously diagnosed by PCR as a normal female and male. Two mosaics were also detected. CONCLUSION: Polymerase chain reaction and fluorescence in situ hybridization are possible for preimplantation sex determination in cases of genetic sex-linked disease. Fluorescence in situ hybridization, however, supplies additional information about sex chromosome aneuploidy and is not susceptible to contamination or misdiagnosis of monosomy X
— id: 66622, year: 1994, vol: 61, page: 111, stat: Journal Article,

Ectopic pregnancies after in vitro fertilization and embryo transfer
Pyrgiotis E; Sultan KM; Neal GS; Liu HC; Grifo JA; Rosenwaks Z
1994 Feb;11(2):79-84, Journal of assisted reproduction & genetics
OBJECTIVE: Our objective was to analyze the risk factors, stimulation characteristics, and future fecundity of patients with ectopic pregnancies after in vitro fertilization (IVF). METHODS: We retrospectively evaluated all cases of ectopic pregnancy occurring between January 1989 and March 1993 (Cornell series 1 to 17). A case-control group of intrauterine pregnancies was used for comparison of the stimulation and transfer characteristics. RESULTS: Twenty-seven of 1123 pregnancies (2.4%) were ectopic, following 2812 fresh IVF embryo transfers, while 8 of 105 pregnancies (7.6%) were ectopic, following 405 frozen-thawed embryo transfers. Tubal factor was the cause of infertility in the majority (85.7%) of ectopic pregnancies. No difference was found between the ectopics and the matched controls in stimulation and transfer characteristics. Thirty ectopic pregnancies were ampullary, two were interstitial, two were cervical, and one was heterotopic. Twenty of the patients subsequently underwent 29 IVF attempts, with a pregnancy rate of 41.4% per transfer. CONCLUSIONS: Ectopic pregnancy after IVF appears to be related to preexisting tubal pathology; embryo transfer of cryopreserved thawed embryos in a natural cycle may result in a higher ectopic rate in these patients; in subsequent IVF cycles the intrauterine pregnancy rate of these patients is not decreased
— id: 20783, year: 1994, vol: 11, page: 79, stat: Journal Article,

The parental origin of the distal pronucleus in dispermic human zygotes
Tang YX; Munne S; Reing A; Schattman G; Grifo J; Cohen J
1994 Feb;2(1):79-85, Zygote
The purpose of this investigation was to determine the parental origin of the pronucleus furthest from the second polar body (the distal pronucleus) in dispermic human zygotes. Intact dispermic embryos (n = 53) and those from which the distal pronucleus (n = 50) was removed at the zygote stage were biopsied after cleavage. Blastomeres were sexed using either coamplification of X and Y probes using a duplex polymerase chain reaction (PCR), or simultaneous fluorescence in situ hybridisation (FISH) with directly fluorochrome-labelled probes for chromosomes X, Y and 18. The ratio X/Y was determined in both groups of embryos by assessing a minimum of two blastomeres. If the pronuclei in dispermic zygotes are topographically in a fixed position, the X/Y ratio should change from 1:3 in dispermic embryos to 1:1 in enucleated ones. The ratio of embryos containing only an X chromosome and those with X as well as Y chromosomes in the intact dispermic zygotes was 1.0:2.3 which is similar to the theoretical ratio of 1:3. This ratio was 1.0:1.3 in dispermic zygotes from which the distal pronuclei were removed. This ratio is not significantly different from the 1:1 ratio based on a statistical analysis with a sample size of 50. These sex ratios would have been considered different if more than 200 enucleations had been performed. Although the ratio X/Y was altered following removal of distal pronuclei, suggesting frequent targeting of male pronuclei, accidental removal of the female pronucleus could not be excluded. This indicates that enucleation of dispermic zygotes could produce high yields of gynogenetic and androgenetic embryos for research purposes. Clinical application aimed at producing biparental zygotes may be hazardous, since mosaicism was common among enucleated embryos
— id: 66621, year: 1994, vol: 2, page: 79, stat: Journal Article,

Unsuspected Chlamydia trachomatis infection and in vitro fertilization outcome
Witkin SS; Sultan KM; Neal GS; Jeremias J; Grifo JA; Rosenwaks Z
1994 Nov;171(5):1208-1214, American journal of obstetrics & gynecology
OBJECTIVE: Chlamydia trachomatis infections of the female genital tract, although a major cause of infertility, are often asymptomatic and undetected. Since many infertile women now seek in vitro fertilization, a procedure whereby fertilization and embryo implantation are precisely timed, we sought to determine the relation between an unsuspected C. trachomatis infection and the ability of embryos to implant and develop after their transfer to the uterus. STUDY DESIGN: At the time of oocyte aspiration, endocervical samples were obtained from 216 women and assayed by enzyme-linked immunoassay for immunoglobulin A antibodies to C. trachomatis structural membrane components and to recombinant C. trachomatis heat shock protein. The presence of C. trachomatis in the cervices was assessed by the polymerase chain reaction. The outcome of each in vitro fertilization cycle was then ascertained. RESULTS: Oocytes from 198 (91.7%) of the women were fertilized in vitro and subsequently transferred to the uterus. Term deliveries of healthy infants occurred after 68 (34.3%) of these transfers. Cervical immunoglobulin A antibodies to chlamydial heat shock protein were detected in 5 (7.3%) of the women with term births, and 1 (1.5%) also had immunoglobulin A antibody to chlamydial structural components; 3 (4.4%) were positive by the polymerase chain reaction for C. trachomatis. In contrast, among the 130 women whose embryo transfers did not result in an ongoing pregnancy, 36 (27.7%) had cervical antiheat shock protein immunoglobulin A (p = 0.0007) and 24 (18.5%) had antichlamydial structural component immunoglobulin A (p = 0.0002); 15 (11.5%) of these women had positive results of polymerase chain reaction for C. trachomatis. The majority of women with cervical antibodies to chlamydial structural antigens were also positive for antibody to heat shock protein. However, only 35% of the women with antibodies to heat shock protein were also positive for the other chlamydial antibodies. C. trachomatis was detected by polymerase chain reaction in 29.2% of women with anti-C. trachomatis antibodies and 7.8% of women with anti-heat shock protein antibodies. Women positive for antichlamydial immunoglobulin A were more likely to be undergoing a repeat in vitro fertilization cycle than were women who were antibody negative (p = 0.007). CONCLUSION: Unsuspected C. trachomatis infection or reactivation of an immune response to the C. trachomatis heat shock protein may induce an inflammatory reaction in the uterus that impairs embryo implantation and/or facilitates immune rejection after uterine transfer of in vitro fertilized embryos
— id: 20782, year: 1994, vol: 171, page: 1208, stat: Journal Article,

Origin of single pronucleated human zygotes
Munne S; Tang YX; Grifo J; Cohen J
1993 May;10(4):276-279, Journal of assisted reproduction & genetics
PURPOSE: Diploidy in embryos developing from single pronucleated zygotes can occur following parthenogenetic activation or by asynchronous inflation of pronuclei following fertilization. To distinguish between these two mechanisms, sexing was performed. RESULTS: The presence of a Y chromosome in the embryo was considered proof that fertilization had occurred. Twenty-one dividing embryos originating from single pronucleated zygotes were analyzed using the polymerase chain reaction or fluorescence in situ hybridization. In total, 43% (9/21) of the embryos showed Y chromosomes. CONCLUSION: It can be extrapolated that more than 80% of them originated from fertilized eggs
— id: 66623, year: 1993, vol: 10, page: 276, stat: Journal Article,

Sex distribution in arrested precompacted human embryos
Munne S; Tang YX; Weier HU; Stein J; Finkelstein M; Grifo J; Cohen J
1993 May;1(2):155-162, Zygote
Evidence of sexual dimorphism before fetal gonadal differentiation in mammals has been accumulating, suggesting that male embryos develop faster than female ones. The current investigation was performed to evaluate whether the development rate of precompacted human embryos is controlled by sex chromosomes. Sex was determined by polymerase chain reaction and fluorescence in situ hybridisation in 172 arrested embryos derived from in vitro fertilisation. The sex ratio (1.02:0.98) did not differ significantly from 1:1. Although more males appeared to have greater fragmentation, the difference between the sex ratios of highly fragmented and normal embryos (1.08:0.92) was not significant. Arrested female embryos had a tendency to exhibit more than five nuclei and less than 10% fragmentation, but the trend was not statistically significant. The current results suggest that the first developmental block in human embryos occurs prior to and shortly after genomic activation and is not determined by the presence of the Y chromosome
— id: 66624, year: 1993, vol: 1, page: 155, stat: Journal Article,

Detection of Chlamydia trachomatis in semen by the polymerase chain reaction in male members of infertile couples
Witkin, S S; Jeremias, J; Grifo, J A; Ledger, W J
1993 May;168(5):1457-1462, American journal of obstetrics & gynecology
OBJECTIVE: Our objective was to evaluate the presence of asymptomatic Chlamydia trachomatis infection by means of the polymerase chain reaction in male members of couples with previously undiagnosed infertility. STUDY DESIGN: Twenty-eight infertile-couples who had negative cultures or negative results when tested by deoxyribonucleic acid probe for Chlamydia trachomatis in semen and cervical samples were studied. Semen samples were tested for Chlamydia trachomatis by means of the polymerase chain reaction. Sera from both partners were diluted 1:128 and tested for immunoglobulin M antibodies to Chlamydia. Sera and ejaculated sperm were evaluated for the presence of antisperm antibodies. Semen analyses were also performed. RESULTS: Chlamydia trachomatis was identified in semen from 11 (39.3%) of the male partners. Its detection correlated with the presence in the ejaculate of motile sperm containing antisperm antibodies (p < 0.01). Either antisperm immunoglobulin G, immunoglobulin A, or both were located on sperm only from 5 (45.5%) of the 11 men whose results were positive when tested for Chlamydia trachomatis. Similarly, immunoglobulin G or immunoglobulin A antibodies to sperm were only detected in 5 (45.5%) of the spouses of men with Chlamydia in semen. Immunoglobulin M antibody to Chlamydia trachomatis was identified in only one of the men. However, antichlamydial immunoglobulin M antibodies were present in sera from 6 (54.5%) female partners of men with seminal Chlamydia trachomatis but in none of the other 17 women (p < 0.01). CONCLUSION: Although undetected by culture of deoxyribonucleic acid probe of semen samples, Chlamydia trachomatis was nevertheless identified in semen of some symptom-free men by the polymerase chain reaction. This is probably a result of the increased sensitivity of the polymerase chain reaction to detect Chlamydia trachomatis. The increased prevalence of an autoimmune response to sperm in men with this organism in their semen suggests that a subclinical chlamydial infection may activate an immune response to sperm. A similar association between Chlamydia trachomatis in semen and circulating antisperm antibodies in female partners indicates that Chlamydia may also induce an immune response to sperm in women. Infertility in these couples may be the result of a direct inflammatory response in the cervix or endometrium to repeated Chlamydia exposure or of the ability of Chlamydia to evoke an immune response to spermatozoa
— id: 120781, year: 1993, vol: 168, page: 1457, stat: Journal Article,

Primer extension preamplification for detection of multiple genetic loci from single human blastomeres
Xu, K; Tang, Y; Grifo, J A; Rosenwaks, Z; Cohen, J
1993 Dec;8(12):2206-2210, Human reproduction
A new technology called primer extension preamplification (PEP), which has been applied to single spermatozoa, increases the amount of polymerase chain reaction (PCR) templates by amplifying DNA of the whole genome. The current investigation was aimed at applying PEP to single human blastomeres. Two blastomeres with nuclei from arrested embryos were selected for this study. Using three different PEP protocols (experiments I, II and III), DNA from single blastomeres was amplified using 15-base oligonucleotide random primers. The efficiency of the procedure was determined by further amplifications of aliquots of the PEP products with two specific sequences. Three aliquots from each PEP product were used as PCR templates for the human X chromosome (X) or the exon 10 of the cystic fibrosis gene (CF). PCR amplified products were analysed by gel electrophoresis. In experiment I, when X primers were used, positive signals were detected in all 10 embryos (100%), 90.0% (18/20) of the blastomeres, and in 80.0% (96/120) of the replicates. When CF primers were amplified, all embryos (100%, 10/10), 90.9% (18/20) of the blastomeres and 78.3% (47/60) of the replicates were positive. In experiment II, efficiency was significantly reduced when total time for the procedure was minimized from 8 h to 5 h and 45 min. Although the time was further reduced to 4 h and 40 min in experiment III, the efficiency remained the same as in experiment I when the volume of PEP was reduced from 60 microliters (experiments I and II) to 40 microliters. One out of 132 control replicates (0.8%) was contaminated.(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 120780, year: 1993, vol: 8, page: 2206, stat: Journal Article,

Microsurgical fertilization procedures: the absence of stringent criteria for patient selection
Cohen, J; Alikani, M; Adler, A; Berkeley, A; Davis, O; Ferrara, T A; Graf, M; Grifo, J; Liu, H C; Malter, H E
1992 Jun;9(3):197-206, Journal of assisted reproduction & genetics
Subzonal sperm insertion and partial zona dissection were applied in 250 in vitro fertilization cycles in couples (n = 200) with abnormal semen analyses; 61 clinical pregnancies were established (24% per egg retrieval). Patients were selected without using minimal cutoff criteria. The study included patients with 0% normal sperm forms (strict criteria), no motile sperm (but some live cells), and sperm counts which could be assessed only after centrifugation. Patients were categorized into three subsets. Group A (n = 116 cycles) failed to fertilize in a previous cycle. Group B (n = 40) was excluded from IVF due to the severity of sperm profiles, such as a maximum of 2% normal forms. Group C (n = 94) constitutes those patients for whom a standard cycle could possibly result in failure. Monospermic fertilization rates were 18% (A), 19% (B), and 24% (C). The incidences of embryo replacement were 63% (A), 53% (B), and 69% (C). Rates of clinical pregnancy were 22% (A), 23% (B), and 28% (C). The presence of one, two, or three semen abnormalities did not correlate with the outcome of microsurgical fertilization. Twenty-two percent of patients with combined oligoasthenoteratozoospermia became pregnant. Moreover, ongoing pregnancies were established in instances with 0% normal sperm forms and no progressively motile spermatozoa. It is concluded that stringent cutoff criteria may not be necessary when both partial zona dissection and subzonal sperm insertion are performed efficiently
— id: 130973, year: 1992, vol: 9, page: 197, stat: Journal Article,

Pregnancy after embryo biopsy and coamplification of DNA from X and Y chromosomes
Grifo JA; Tang YX; Cohen J; Gilbert F; Sanyal MK; Rosenwaks Z
1992 Aug 12;268(6):727-729, JAMA
— id: 66625, year: 1992, vol: 268, page: 727, stat: Journal Article,

Preimplantation genetic diagnosis. In situ hybridization as a tool for analysis
Grifo, J A; Boyle, A; Tang, Y X; Ward, D C
1992 Apr;116(4):393-397, Archives of pathology & laboratory medicine
In situ hybridization allows one to directly visualize DNA sequences of interest for which probes are available. Fluorescent signals can be detected in single cells in either metaphase or interphase nuclei, thus obviating the need for cell synchronization. The potential to use this technology in conjunction with in vitro fertilization techniques to genetically analyze an embryo before transfer could offer patients at genetic risk an alternative. Currently, these patients wait until 9 to 16 weeks' gestation to find out about the genetic makeup of their pregnancy. As such, sexing the embryos of patients at risk for transmitting sex-linked disorders or for performing numerical chromosome analysis could be performed before embryo transfer. Techniques for applying in situ hybridization to single cells obtained from embryos are presented. Hybridization of specific probes to blastomeres of mouse and human embryos as well as sperm is demonstrated
— id: 120784, year: 1992, vol: 116, page: 393, stat: Journal Article,

Evaluation of Vero cell co-culture system for mouse embryos in various media
Lai YM; Stein DE; Soong YK; Tang YX; Grifo J; Malter HE; Talansky BE; Cohen J; Liu HC; Rosenwaks Z
1992 Feb;7(2):276-280, Human reproduction
Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embryos with Vero cells in both Dulbecco's modified Eagle's medium/Ham's F12 (DMEM/F12) and human tubal fluid (HTF) culture medium. Two-cell CB6F1 mouse embryos were cultured either in the presence of Vero cells (group A) or in culture medium alone (group B). In DMEM/F12 significantly more morulae developed in group A than in group B on day 3 (91 versus 23%; P less than 0.01). In contrast, the mouse embryos grew rapidly in HTF and significant differences were noted only in later embryonic stages (on day 5; 86% and 50%, P less than 0.01 of group A and B respectively, hatching or hatched). Similar experiments using DF1 and ICR mouse strains also revealed enhanced embryo development in the presence of Vero cells. To determine whether the embryo-enhancing effects of Vero cells were due to the removal of toxins or to the secretion of embryotrophic factors, ICR mouse embryos were cultured in fresh media with cells (group A), without cells (group B) and in cell-free conditions using cell-conditioned media which were obtained in the presence (group C) or absence (group D) of embryos. These results demonstrated that completion of hatching was highest (52%; P less than 0.01) in group A after 6 days in culture. There were no significant differences between groups B, C and D (rates of total hatching 18, 17 and 17%, respectively). It is concluded that Vero cells improve the development of mouse embryos and this is likely to be due to removal of substances inhibitory to development.(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 66626, year: 1992, vol: 7, page: 276, stat: Journal Article,

Relation between antibodies to Chlamydia trachomatis and spontaneous abortion following in vitro fertilization
Licciardi, F; Grifo, J A; Rosenwaks, Z; Witkin, S S
1992 Jun;9(3):207-210, Journal of assisted reproduction & genetics
BACKGROUND: Many couples undergo in vitro fertilization due to occlusion of the fallopian tubes. Chlamydia trachomatis infections are a major cause of this tubal damage. Since this organism has also been associated with poor pregnancy outcome, we investigated whether a past exposure to C. trachomatis was associated with spontaneous abortion following in vitro fertilization and embryo transfer. METHODS: Sera from 145 women undergoing IVF were diluted 1:128 and tested for IgG antibodies to C. trachomatis by an immunoperoxidase assay, using infected cells fixed to slides. All subjects and their partners were negative for C. trachomatis by culture or by DNA hybridization. RESULTS: Serological evidence of a past chlamydial infection was observed in 33.8% of the women. The incidence of antichlamydial IgG was greater (P less than 0.001) in women whose infertility was due to known tubal disease (37 of 78; 47.4%) than in women whose infertility was due to other causes (12 of 67; 17.9%). Spontaneous abortions after embryo transfer occurred in 20% of the subjects. The incidence of antichlamydial IgG in aborting women (20 of 29; 69.0%) was greater (P less than 0.001) than the incidence in either women with successful pregnancies (9 of 38; 23.7%) or women who did not become pregnant (20 of 78; 25.6%) after IVF. No relation was observed between antichlamydial antibody status and maternal age, the number of oocytes aspirated, the number of oocytes fertilized, and the number of embryos transferred. CONCLUSIONS: A previous infection with C. trachomatis may increase susceptibility to subsequent spontaneous abortion, even in the absence of a detectable current infection
— id: 120783, year: 1992, vol: 9, page: 207, stat: Journal Article,

Aldose reductase inhibition prevents galactose-induced ovarian dysfunction in the Sprague-Dawley rat
Meyer, W R; Doyle, M B; Grifo, J A; Lipetz, K J; Oates, P J; DeCherney, A H; Diamond, M P
1992 Dec;167(6):1837-1843, American journal of obstetrics & gynecology
OBJECTIVE: Our objective was to determine whether impaired ovarian function induced by short-term creation of a galactosemic state in the rat might be prevented by the coadministration of an aldose reductase inhibitor. STUDY DESIGN: Prepubertal Sprague-Dawley rats were fed four different diets including (1) control, (2) 40% galactose, (3) 40% galactose and an aldose reductase inhibitor, and (4) an aldose reductase inhibitor with the control diet. Percentage germinal vesicle breakdown, postovulatory oocyte quantities, hormonal parameters, ovarian histologic evaluation, and ovarian galactitol concentrations were determined. RESULTS: The galactose-fed animals (group 2) had decreased germinal vesicle breakdown (47%) versus control (69%, p < 0.05). Galactose-exposed animals had significantly decreased quantities of postovulatory eggs (6.4 per animal) after menotropin ovarian stimulation in comparison with controls (14.1, p < 0.01). In rats exposed to high dietary levels of galactose (group 2) ovarian galactitol concentrations were significantly higher (protein 42.12 mumol/gm versus 0.0 for controls, p < 0.005). When galactose-fed animals received the aldose reductase inhibitor, ovarian accumulation of galactitol was significantly reduced and the observed detrimental effects on the oocyte were prevented. CONCLUSION: Galactitol accumulation or metabolic flux through aldose reductase in galactosemic rodents may be involved in the demonstrated ovarian dysfunction
— id: 120782, year: 1992, vol: 167, page: 1837, stat: Journal Article,

Baseline ovarian cysts do not affect clinical response to controlled ovarian hyperstimulation for in vitro fertilization
Penzias, A S; Jones, E E; Seifer, D B; Grifo, J A; Thatcher, S S; DeCherney, A H
1992 May;57(5):1017-1021, Fertility & sterility
OBJECTIVE: To evaluate the effect of baseline ovarian cysts at the onset of controlled ovarian hyperstimulation for in vitro fertilization (IVF) on cycle outcome. DESIGN, PATIENTS: A review of 82 IVF cycles in 29 women in which each patient served as her own control. The stimulation regimen for each patient remained constant over time. Each woman had at least one cycle in which an ovarian cyst measuring 14 to 53 mm was present at baseline and one cycle in which no such cyst was present. SETTING: The In Vitro Fertilization Program at Yale University School of Medicine. RESULTS: There was no statistically significant difference in cycle cancellation rates, baseline serum estradiol (E2), peak serum E2, number of follicles present at retrieval, number of oocytes retrieved, or fertilization rate between groups. Stimulation regimen, cyst size, and age were unrelated to outcome. The number of cysts present at baseline correlated positively with the number of follicles present at retrieval. CONCLUSION: Baseline ovarian cysts in the setting of a low baseline E2 level do not affect the clinical response to controlled ovarian hyperstimulation in IVF cycles
— id: 114293, year: 1992, vol: 57, page: 1017, stat: Journal Article,

Preembryo biopsy and analysis of blastomeres by in situ hybridization
Grifo, J A; Boyle, A; Fischer, E; Lavy, G; DeCherney, A H; Ward, D C; Sanyal, M K
1990 Dec;163(6 Pt 1):2013-2019, American journal of obstetrics & gynecology
We developed a method for the biopsy of preimplantation mouse embryos (preembryos) at the four- to eight-cell stage, which uses partial zona pellucida dissection. The preembryos were collected in calcium- and magnesium-free phosphate-buffered saline solution with 0.01% ethylenediaminetetraacetic acid, 0.1 mol/L sucrose, and 4 mg/ml of bovine serum albumin to facilitate removal of blastomeres. This allows entry of a fine micropipette into the perivitelline cavity with subsequent removal of a single blastomere by gentle suction. The majority of embryos (75%) from which biopsy specimens were obtained in this fashion developed to the blastocyst stage. The blastomeres obtained were mainly intact and they were fixed to glass slides. After permeabilization, in situ hybridization was performed with chromosome X- and chromosome 3-specific probes. Human unfertilized eggs and blastomeres from human polyspermic embryos also have been analyzed by in situ hybridization with chromosome specific probes. The combination of nondestructive embryo biopsy and in situ hybridization is a possible approach for preimplantation genetic diagnosis
— id: 120785, year: 1990, vol: 163, page: 2013, stat: Journal Article,

Term interstitial pregnancy with uterine torsion: sonographic, pathologic, and clinical findings
Bond, A L; Grifo, J A; Chervenak, F A; Kramer, E E; Harris, M A
1989 May;73(5 Pt 2):857-859, Obstetrics & gynecology
Interstitial pregnancy which resulted in a term, live infant was found in association with pathologic torsion of the uterus. Antenatal sonograms revealed a hypervascular area anterior to the uterus
— id: 120786, year: 1989, vol: 73, page: 857, stat: Journal Article,

Interferon-gamma in the diagnosis and pathogenesis of pelvic inflammatory disease
Grifo, J A; Jeremias, J; Ledger, W J; Witkin, S S
1989 Jan;160(1):26-31, American journal of obstetrics & gynecology
Serologic markers were evaluated to determine if they could aid in the differential diagnosis of pelvic inflammatory disease in 48 consecutive women seeking evaluation for pelvic pain. On the basis of clinical and microbiologic parameters, 29 patients (60.4%) were diagnosed as having pelvic inflammatory disease. Neisseria gonorrhoeae only was isolated from the cervix of eight (27.6%) patients with pelvic inflammatory disease, five (17.2%) had only Chlamydia, and two (6.9%) had Neisseria and Chlamydia, whereas in 15 (48.3%) patients no pathogen was isolated. Interferon-gamma was present in significantly more sera (p less than 0.025) from patients with pelvic inflammatory disease (65.5%) than from women without pelvic inflammatory disease (15.8%). Sera from 10 healthy women lacked detectable interferon-gamma. In patients with only Neisseria, seven (87.5%) had circulating interferon-gamma; three (60%) of the women with only Chlamydia, one (50%) woman with Neisseria and Chlamydia, and eight (57.1%) with no identified pathogens were also positive for interferon-gamma. Sera from 11 of 28 patients with pelvic inflammatory disease (39%) but only one of 19 sera from women without pelvic inflammatory disease (5%) also inhibited the Candida-induced proliferation of control lymphocytes. This immunosuppressive activity was prevented by immunoprecipitation of interferon-gamma by anti-interferon-gamma antibody but not by treatment with anti-interferon-alpha antibody. The persistence of interferon-gamma in the sera of patients with pelvic inflammatory disease may aid in the differential diagnosis of this disease and increase our understanding of the pathogenesis of microbial-mediated tubal damage
— id: 120787, year: 1989, vol: 160, page: 26, stat: Journal Article,

Influence of 5' proximal secondary structure on the translational efficiency of eukaryotic mRNAs and on their interaction with initiation factors
Lawson, T G; Ray, B K; Dodds, J T; Grifo, J A; Abramson, R D; Merrick, W C; Betsch, D F; Weith, H L; Thach, R E
1986 Oct 25;261(30):13979-13989, Journal of biological chemistry
The effects of 5' proximal secondary structure in mRNA molecules on their translation and on their interaction with the eukaryotic initiation factors (eIF)-4F, eIF-4A, and eIF-4B have been examined. Secondary structures were generated in the 5' noncoding region of rabbit globin and reovirus mRNAs by means of hybridization with cDNA molecules. cDNAs hybridized to the first 15 bases downstream from the cap inhibited the translation of the mRNAs in both reticulocyte and wheat germ lysates. The degree of inhibition was directly related to the monovalent ion concentration and inversely related to reaction temperature. These hybrid structures also reduced the competitive ability of the messages. Hybrid structures beginning downstream from the first 15 bases did not inhibit the translation of beta-globin mRNA or reovirus s3 mRNA. None of the hybrid structures were detrimental to the interaction of the mRNAs with the 26-kDa cap binding protein of eIF-4F, as determined by chemical cross-linking assays. However, in the presence of ATP, hybrid structures immediately adjacent to the cap severely inhibited the cross-linking to the p46 subunit of eIF-4F or to additional eIF-4A or eIF-4B. In order to account for these observations, a two-step mechanism is proposed for the interaction of eIF-4F with the 5' end of an mRNA molecule. The first step involves a weak initial interaction of the p26 subunit with the cap. The second step requires the hydrolysis of ATP and results in the formation of a stable initiation factor-mRNA complex, which may involve eIF-4A and eIF-4B. This second step is inhibited by the presence of 5' proximal secondary structure. In any event, our results demonstrate that the effect of mRNA structure on translation rate depends strongly on its position with respect to the 5' end and that this effect is due at least in part to an inhibition of the action of initiation factors normally required for the unwinding of structure
— id: 120788, year: 1986, vol: 261, page: 13979, stat: Journal Article,

Shutoff of host translation by encephalomyocarditis virus infection does not involve cleavage of the eucaryotic initiation factor 4F polypeptide that accompanies poliovirus infection
Mosenkis, J; Daniels-McQueen, S; Janovec, S; Duncan, R; Hershey, J W; Grifo, J A; Merrick, W C; Thach, R E
1985 May;54(2):643-645, Journal of virology
Studies were conducted to determine whether encephalomyocarditis virus infection causes proteolytic cleavage of any of the polypeptides which comprise eucaryotic initiation factor 4F. Since no such alterations in the components of the initiation factor were detected, these observations confirmed that the mechanisms whereby encephalomyocarditis virus and poliovirus shut off host translation are different
— id: 120790, year: 1985, vol: 54, page: 643, stat: Journal Article,

ATP-dependent unwinding of messenger RNA structure by eukaryotic initiation factors
Ray, B K; Lawson, T G; Kramer, J C; Cladaras, M H; Grifo, J A; Abramson, R D; Merrick, W C; Thach, R E
1985 Jun 25;260(12):7651-7658, Journal of biological chemistry
Interaction of protein synthesis initiation factors with mRNA has been studied in order to characterize early events in the eukaryotic translation pathway. Individual reovirus mRNAs labeled with 32P in the alpha position relative to the m7G cap and eukaryotic initiation factor (eIF)-4A, -4B, and -4F purified from rabbit reticulocytes were employed. It was found that eIF-4A causes a structural change in mRNA, as evidenced by a nuclease sensitivity test: addition of high concentrations of eIF-4A greatly increase the nuclease sensitivity of the mRNA, suggesting that this factor can melt or 'unwind' mRNA structure. ATP is required for this reaction. At low concentrations of eIF-4A, addition of eIF-4B is required for maximal unwinding activity. Thus eIF-4B enhances eIF-4A activity. Addition of eIF-4F also makes the mRNA sensitive to nuclease indicating a similar unwinding role to that of eIF-4A. Stoichiometric comparisons indicate that eIF-4F is more than 20-fold more efficient than eIF-4A in catalyzing this reaction. The unwinding activity of eIF-4F is inhibited by m7GDP, while that of eIF-4A is not. This suggests that eIF-4A functions independent of the 5' cap structure. Our results also suggest that the unwinding activity of eIF-4F is located in the 46,000-dalton polypeptide of this complex, which has shown by others to be similar or identical to eIF-4A
— id: 120789, year: 1985, vol: 260, page: 7651, stat: Journal Article,

RNA-stimulated ATPase activity of eukaryotic initiation factors
Grifo, J A; Abramson, R D; Satler, C A; Merrick, W C
1984 Jul 10;259(13):8648-8654, Journal of biological chemistry
Previously, we have described an ATP-dependent recognition and binding of mRNA by eukaryotic initiation factors (eIF)-4A, eIF-4B, and eIF-4F (Grifo, J. A., Tahara, S. M., Leis, J. P., Morgan, M. A., Shatkin, A. J., and Merrick, W. C. (1982) J. Biol. Chem. 257, 5246-5252; Grifo, J. A., Tahara, S. M., Morgan, M. A., Shatkin, A. J., and Merrick, W. C. (1983) J. Biol. Chem. 258, 5804-5810). This finding was consistent with other studies which implicated eIF-4A and eIF-4B in binding mRNA to the 40 S ribosomal subunit, an ATP-requiring process. As part of ongoing studies of this step, and, in particular its ATP requirement, we have examined ATPase activity of various initiation factors. In this communication we describe an RNA-dependent ATP hydrolysis catalyzed by eIF-4A and eIF-4F. Although eIF-4B has little or no ATPase activity it can stimulate the RNA-dependent ATPase activity of either eIF-4A or eIF-4F. Similar to the ATP-dependent mRNA binding assay, the RNA-dependent ATPase activity is inhibited by the cap analogue m7GDP when globin mRNA is used as the activator. In addition, a variety of polynucleotides stimulate the ATPase activity of these factors including rRNA, tRNA, poly(U), and poly(A) but not poly(dA). Finally, an attempt has been made to discern whether phosphorylation or ATP hydrolysis is responsible for the ATP-stimulated binding of mRNA by eIF-4A and eIF-4B. We present evidence which is consistent with the interpretation that ATP hydrolysis and not protein phosphorylation correlates with ATP-stimulated binding of mRNA
— id: 120791, year: 1984, vol: 259, page: 8648, stat: Journal Article,

Unusual requirements for optimum translation of polio viral RNA in vitro
Daniels-McQueen, S; Detjen, B M; Grifo, J A; Merrick, W C; Thach, R E
1983 Jun 10;258(11):7195-7199, Journal of biological chemistry
The translation of poliovirion RNA (polio RNA) in an in vitro fractionated system was much less efficient than that of encephalomyocarditis virion RNA (EMC RNA). In contrast, when polio and EMC RNAs were added to postmitochondrial cell lysates (S10), they were translated with equal efficiency. However, this equality was observed only when high concentrations of S10 were employed; at lower concentrations, polio RNA translation was reduced relative to that of EMC RNA. These results suggest that both the fractionated and S10 systems are limiting in a component that is required for the optimal translation of polio RNA. The elongation rates for EMC and polio RNA translation in the fractionated system were found to be similar, indicating that this component acts at an initiation step. Various components, including excess ribosomal salt wash and postribosomal supernatant of cell lysate, were added to the fractionated system in an effort to identify the slow step more precisely. Of these, only excess ribosomal salt wash specifically stimulated polio RNA translation, suggesting that one or more initiation factors is necessary in unusually large amounts for this mRNA. Various purified initiation factors were tested for the ability to enhance polio RNA translation. Of these, only purified eukaryotic initiation factor 4A had a specific effect. This suggests that polio RNA, in contrast to other mRNAs tested (EMC, reoviral, and globin), may have an unusually low affinity for this initiation factor. The significance of these results is discussed in terms of the methods picornaviruses have evolved for reprogramming the translational machinery of the host cell
— id: 120793, year: 1983, vol: 258, page: 7195, stat: Journal Article,

New initiation factor activity required for globin mRNA translation
Grifo, J A; Tahara, S M; Morgan, M A; Shatkin, A J; Merrick, W C
1983 May 10;258(9):5804-5810, Journal of biological chemistry
A reconstituted reticulocyte translation system originally designed to be deficient in eukaryotic initiation factor 4B (eIF-4B) was used to identify a new activity required for maximal synthesis of rabbit globin. This new activity purifies as a stable, high molecular weight complex by a variety of chromatographic procedures and is termed eIF-4F. The purified globin stimulatory activity also restores translation of capped mRNAs in extracts of poliovirus-infected HeLa cells. Like restoring activity that was obtained as a protein complex by different procedures (Tahara, S. M., Morgan, M. A. and Shatkin, A. J. (1981) J. Biol. Chem. 256, 791-794), eIF-4F includes the 24,000-dalton cap binding protein and major polypeptides of Mr approximately 200,000 and approximately 46,000. The latter component comigrates with eIF-4A by two-dimensional gel electrophoresis and, like eIF-4A, chemically cross-links to the 5'-end of capped mRNA by an ATP-dependent, m7GDP-sensitive reaction. Unlike eIF-4F, cap binding protein of Mr approximately 24,000 isolated by affinity chromatography on m7GDP-Sepharose does not stimulate globin synthesis in the reconstituted system
— id: 120794, year: 1983, vol: 258, page: 5804, stat: Journal Article,

Role of mRNA competition in regulating translation: further characterization of mRNA discriminatory initiation factors
Ray, B K; Brendler, T G; Adya, S; Daniels-McQueen, S; Miller, J K; Hershey, J W; Grifo, J A; Merrick, W C; Thach, R E
1983 Feb;80(3):663-667, Proceedings of the National Academy of Sciences of the United States of America
Host and reovirus mRNAs compete with one another for translation in infected cells. Kinetic analysis has suggested that the site of competition is a message discriminatory initiation factor which must bind to the mRNA before it can interact with the 40S ribosomal subunit. The present communication describes an in vitro assay which can detect message discriminatory activities. A competitive situation is established by using reovirus and globin mRNAs, and then the specificity with which this competition is relieved by added components is measured. Among the various initiation factors surveyed with this assay, two have the properties expected of the mRNA discriminatory factor. These are eukaryotic initiation factor 4A and a 'cap binding protein' complex. Inasmuch as the cap binding protein complex contains a subunit similar or identical to the initiation factor eIF-4A, it seems likely that only one form of the latter factor may be active in vivo. In vitro, both factors relieve competition among both capped and uncapped reovirus mRNAs according to similar hierarchies. These results suggest that some feature other than the m7G cap, such as nucleotide sequence or secondary structure, is recognized by the discriminatory factor
— id: 120795, year: 1983, vol: 80, page: 663, stat: Journal Article,

Phospholipid-sensitive Ca2+-dependent protein kinase phosphorylates the beta subunit of eukaryotic initiation factor 2 (eIF-2)
Schatzman, R C; Grifo, J A; Merrick, W C; Kuo, J F
1983 Aug 8;159(1-2):167-170, FEBS letters
The ability of homogeneous phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) from pig spleen to phosphorylate eukaryotic initiation factor 2 (eIF-2) was examined. PL-Ca-PK phosphorylated the beta-subunit of eIF-2, whereas myosin light chain kinase (MLCK) and cyclic AMP- and cyclic GMP-dependent protein kinases (cA-PK and cG-PK) did not. PL-Ca-PK could incorporate a maximum of 1.6 mol phosphate/mol eIF-2. The app. Km and Vmax for PL-Ca-PK phosphorylation of eIF-2 were 0.13 microM and 0.02 mumol.min-1.mg enzyme-1, respectively. Phosphoamino acid analysis revealed that incorporation of phosphate into eIF-2 occurred almost exclusively at serine residues. These findings indicate that eIF-2 was an effective substrate for PL-Ca-PK, suggesting that this enzyme may play a role in the regulation of protein synthesis
— id: 120792, year: 1983, vol: 159, page: 167, stat: Journal Article,

Characterization of eukaryotic initiation factor 4A, a protein involved in ATP-dependent binding of globin mRNA
Grifo, J A; Tahara, S M; Leis, J P; Morgan, M A; Shatkin, A J; Merrick, W C
1982 May 10;257(9):5246-5252, Journal of biological chemistry
Eukaryotic initiation factor 4A (eIF-4A) has been purified (to apparent homogeneity) from rabbit reticulocyte lysate. It is a single polypeptide accounting for at least 90% of the Coomassie blue staining material when subjected to gel electrophoresis in the presence of sodium dodecyl sulfate. The molecular weight was determined by gel electrophoresis under denaturing conditions at two different bisacrylamide to acrylamide ratios, by gel filtration under native conditions and by sedimentation equilibrium at three different protein concentrations. Additional physical properties of the polypeptide were also determined. In an attempt to characterize the function of eIF-4A, a protein specifically required for mRNA translation, an assay was developed which measures the protein-dependent retention of radiolabeled hemoglobin mRNA on nitrocellulose filters. These studies led to the discovery of an ATP-stimulated binding of mRNA which is dependent on the presence of eIF-4A and eIF-4B that also contains the 24,000-dalton cap binding protein. The reaction apparently requires ATP hydrolysis since a nonhydrolyzable analogue of ATP, adenosine 5'-(beta, gamma-imino)triphosphate, does not stimulate mRNA binding and GTP cannot substitute for ATP. In addition, ATP-stimulated binding of mRNA can be inhibited by an analog of the mRNA 5' terminus, m7GMP, suggesting recognition of the capped 5' end of hemoglobin mRNA. Consistent with this suggestion, ATP also stimulated the covalent cross-linking of eIF-4A and eIF-4B to the cap of oxidized reovirus mRNA, an interaction that was inhibited by m7GDP
— id: 120796, year: 1982, vol: 257, page: 5246, stat: Journal Article,