Lewis C Krey, Ph.D.

What do young women really want? A look inside the minds of potential oocyte donors
Druckenmiller S.; Knopman J.M.; DeVore S.; Krey L.; Noyes N.
2011 ;26:i121-i122, Human reproduction
Introduction: The number of women pursuing advanced degrees and competitive careers has greatly increased and they now represent 50 + % in many postgraduate programs. This career focus along with the availability of multitudinous contraceptive options has allowed women to postpone starting a family to a convenient time, and the number of women delaying childbearing and their age at first birth (= 25y) is at an all time high. The traditional nuclear family has also been remodeled with regard to composition as increased numbers of single/same-sex parents look to conceive. Reproductive medicine has adapted to accommodate these modern practices and has recently experienced an explosion relative to the interest in/use of donor oocytes (DO). Given this trend, our study's objective was twofold: To assess young women's knowledge regarding their fertility and potential infertility, and to understand what factors motivate them to consider donating their oocytes. Material and Methods: In 2010, an IRB-approved electronic questionnaire (65 multiple-choice questions) was randomly distributed to female undergraduate and graduate students (range:18-32y; mean:21-22y) attending American universities. To date 1,142 responses have been analyzed. Results: The majority of respondents (59%) indicated that fertility was 'important/ very important' to them, but 79% considered themselves to have 'no/ minimal knowledge' about this subject. Only 2% considered themselves 'very knowledgeable'. In fact, when asked 3 basic infertility questions, 80% answered 2 incorrectly. Older respondents and those pursuing advanced degrees most often answered these questions correctly. The majority of respondents ranked preferable conception-enhancing modalities as follows: Natural conception, ART, adoption, DO-anonymous, then DO-directed/known. In fact, 67% stated that if they could not conceive naturally, they would use adoption in lieu of DO. The most commonly cited reason for not pursuing DO-directed/known was resistance to 'bearing a child of someone they know.' When queried about DO, half of the respondents would consider donating for research, to further science (89%) or for financial gain (70%). Those unwilling cited fear of the egg-harvest procedure (45%) and opposition to oocyte experimentation (56%). Only 40% of the respondents indicated that they would donate oocytes for clinical use; 84% wanted to help others create a family, 74% had financial motivation and 67% desired to further science. Interestingly, ~60% of these women indicated they would be more likely to donate if they could control the selection of their recipient(s) according to education/IQ, income, location of oocyte usage, and/or parental health and that they would willingly accept less compensation in exchange for more control of these recipient characteristics. Moreover, 73% desired <3 recipients receiving their oocytes and similar percentages indicated that they would like to know how many oocytes they produced, how many people received their oocytes, if the recipients got pregnant and whether the recipients would be comfortable with their children contacting them at 18-21y. Finally, 43% of pro-DO respondents believe donors should receive additional compensation for desirable characteristics, including education/ IQ, beauty, personality/people skills and athletic competence. Respondents opposed to clinical donation cited they did not want a biological child-of-whom-they-were-unaware/did not raise/fear of how raised. In addition, age, advanced degree, and family income impacted their decision on oocyte donation. Conclusions: While the majority of these young women considered fertility of paramount importance, most were ignorant of basic fertility principles suggesting that the core health/science curricula taught before/at this age are lacking. In conjunction with the worldwide delay in childbearing, it is prudent and timely to implement more effective programs geared toward the reproductive education of young women. Additionally, potential donors yearn for more control over recipient characteristics as well as knowledge of the outcome(s) of their donations. This may become more relevant as demand increases and frozenoocyte banks emerge around the world
— id: 137460, year: 2011, vol: 26, page: i121, 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,

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,

Monozygotic twinning: an eight-year experience at a large IVF center
Knopman, Jaime; Krey, Lewis C; Lee, Jennifer; Fino, Mary Elizabeth; Novetsky, Akiva P; Noyes, Nicole
2010 Jul;94(2):502-510, Fertility & sterility
OBJECTIVE: To characterize incidence, chorionicity, amnionicity, and pregnancy outcome for monozygotic twin pregnancy (MZT) after IVF. DESIGN: Retrospective review. SETTING: University-based fertility center. PATIENT(S): Autologous and oocyte donation IVF cycles eventuating in 4,976 clinical gestations from 2000 to 2007. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): MZT incidence, chorionicity, zygosity, pregnancy outcome. RESULT(S): Ninety-eight MZTs were diagnosed after first-trimester ultrasound evaluation (2% incidence). The incidence in cycles transfering autologous oocytes was 1.7% but was 3.3% with donor oocytes; however, women <35 years old using their own oocytes displayed a similar rate (3.1%) to women using donor oocytes. Eighty MZTs occurred after fresh day-5 transfer; only 14 followed fresh day-3 transfer (2.6% vs. 1.2%). The MZT incidence in day-3 transfers without hatching was not different from those with hatching (1.3% vs. 1.1%). In addition, MZT incidence did not differ significantly whether or not ICSI was performed (2.4% vs. 2.0%). Four MZTs occurred after frozen-thawed embryo transfer (0.8% incidence). Ninety-five percent of all placental arrangements were confirmed as monochorionic-diamniotic on obstetric ultrasounds. CONCLUSION(S): These findings confirm a higher incidence of MZT after IVF. Monochorionic-diamniotic implantations were increased, whereas monochorionic-monoamniotic were not. The MZT risk factors included young age and extended culture, but not zona penetration or cryopreservation
— id: 138161, year: 2010, vol: 94, page: 502, 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,

Evaluating the necessity for universal screening of prospective oocyte donors using enhanced genetic and psychological testing
Reh, Andrea; Amarosa, Alana; Licciardi, Frederick; Krey, Lewis; Berkeley, Alan S; Kump, Lisa
2010 Sep;25(9):2298-2304, Human reproduction
BACKGROUND To minimize the potential for harmful inheritable conditions, donors are rigorously screened according to standard guidelines, yet such guidelines may not be sufficient to exclude egg donors with certain known inheritable conditions. We compared universal screening of oocyte donors with Tay-Sachs, Fragile X, karyotype and Minnesota Multiphasic Personality Inventory-2 (MMPI-2) versus standard American Society of Reproductive Medicine (ASRM) guidelines that do not include such testing. METHODS In this 12 year retrospective cohort study, results of enhanced universal screening of all anonymous oocyte donor candidates from 1997 to 2008 at a university hospital oocyte donation program were reviewed. Primary outcomes were the frequency of oocyte donor candidates excluded as a result of enhanced universal screening (Tay-Sachs, Fragile X, karyotypic analysis and MMPI-2) versus basic screening according to ASRM guidelines. RESULTS Of 1303 candidates who underwent on-site evaluation, 47% passed the screening process, 23% were lost to follow-up and 31% were excluded. Genetic and psychological factors accounted for the most common reasons for candidate exclusion. Enhanced genetic screening excluded an additional 25 candidates (19% of all genetic exclusions) and enhanced psychological screening excluded an additional 15 candidates (12% of all psychological exclusions). Altogether enhanced screening accounted for 40 candidates (10%) of the total pool of excluded candidates. CONCLUSIONS Although our study is limited by its retrospective nature and center-specific conclusions, we show that enhanced comprehensive screening can exclude a significant number of candidates from an oocyte donor program and should be encouraged to assure optimal short-term and long-term outcomes for pregnancies achieved through oocyte donation
— id: 111821, year: 2010, vol: 25, page: 2298, 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,

Are gonadotropin-releasing hormone agonists losing popularity? Current trends at a large fertility center
Reh, Andrea; Krey, Lewis; Noyes, Nicole
2010 Jan;93(1):101-108, Fertility & sterility
OBJECTIVE: To explore the long- and short-term trends in LH-suppression protocol use and patient profile characteristics. DESIGN: Descriptive study, retrospective cohort. SETTING: Large, university-based IVF center. PATIENT(S): Four thousand five hundred one fresh IVF cycles categorized by use of GnRH antagonist, luteal GnRH agonist, and follicular microdose GnRH agonist. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Frequency of use of LH-suppression protocol, patient and cycle characteristics, and outcomes at 10-year (1996-2005), 5-year (2001-5), and 3-year intervals (2004-6). RESULT(S): In both the <40 and >/=40 age groups, GnRH antagonist use increased from 2001 to 2005, while luteal GnRH agonist and microdose use decreased. The most recent luteal agonist patients were better responders and had higher implantation, clinical pregnancy, and delivery rates. Antagonist patients in the <40 and >/=40 age groups had a better response in 2005 than in 2001 with higher clinical pregnancy rates. Microdose patients responded worse in 2005 than in 2001, although pregnancy rates did not change significantly. Such trends were echoed from 2004 to 2006. CONCLUSION(S): The target population for GnRH antagonist has broadened to include younger, normal responders in addition to the traditional poor responder. Luteal agonist and microdose protocols are chosen less frequently and remain targeted toward good and poor responders, respectively
— id: 90875, year: 2010, vol: 93, page: 101, stat: Journal Article,

Does the inter-cycle duration affect clinical outcome for fresh IVF cycles?
Cho M.; Mullin C.; Dutta S.; Krey L.C.; Licciardi F.
2009 ;92(3 SUPPL 1):S78-S78, Fertility & sterility
OBJECTIVE: To determine if there is a difference between clinical outcomes with respect to inter-cycle time period. DESIGN: Retrospective analysis MATERIALS AND METHODS: We analyzed the clinical outcomes of 168 patients who underwent 2 consecutive fresh IVF cycles with various intercycle time duration from 1/2001-3/2009. Inclusion criteria were 2 consecutive IVF cycles in which the first cycle did not result in a pregnancy. All poor prognostic patients, whose medical history was characterized by high FSH levels on day 2 (D2) or a prior poor response to gonadotropins, were excluded. Patients were grouped according to time interval between cycles (grp 0: back to back cycles, grp 1: 35-59 days, grp 2: 60-89 days). Analysis included age, D2 FSH, D2 estradiol (E2), total gonadotropin dose (IU), # of eggs retrieved (ER), # embryos transferred (ET), cancellation rate (CR), no start cycle rate, clinical pregnancy rate (PR) and twin pregnancy rate. CR reflected those patients who did not undergo egg retrieval. No Starts patients did not start a cycle secondary to elevated D2 bloodwork or ovarian cysts. PR included patients with a fetal heartbeat(s) activity on ultrasound. Outcomes of the second IVF cycle were analyzed by chi square and ANOVA. RESULTS: Comparisons of Clinical Outcomes. (Table presented). CONCLUSIONS: Patients opting for back-to-back cycles (grp 0) had a significantly higher likelihood of not starting the second cycle when compared to patients who took a 1 month reprieve. This may be due to a high prevalence of residual ovarian cysts. However, if patients were eligible to start a subsequent cycle and completed the cycle, then there was no statistical difference in PR
— id: 127245, year: 2009, vol: 92, page: S78, 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,

Impact of developmental stage at cryopreservation and transfer on clinical outcome of frozen embryo cycles
Noyes, Nicole; Reh, Andrea; McCaffrey, Caroline; Tan, Orkun; Krey, Lewis
2009 ;19 Suppl 3:9-15, Reproductive biomedicine online
Although several early IVF successes were achieved after transferring fully formed blastocysts, the majority of embryos replaced worldwide over the past 30 years have been at the cleavage stage. The programme at this study centre has previously found that delaying an embryo transfer to day 5 can reduce the chance for a high-order multiple gestation without compromising the pregnancy rate because fewer embryos can be replaced. To evaluate the impact of transfer day and embryonic stage at cryopreservation on cycle outcome, 6069 fresh and 706 frozen transfers from 2000-2006 performed at this study centre were retrospectively analysed. Approximately half of the fresh transfers were performed on day 3, with a shift to day-5 transfer over the study period with no change in cryopreservation incidence. Implantation, clinical pregnancy and live birth rates were significantly higher following day-5 transfer. When frozen-thawed embryos (2-cell to day-6 blastocysts) were transferred, acceptable pregnancy and live birth rates were achieved at all stages but thawed embryos transferred as day-5 blastocysts generated consistently higher clinical pregnancy and live birth rates. Transfer of embryos frozen on day 6 had the highest miscarriage and lowest live birth rates. Barring government regulation, an IVF programme's day for cryopreservation generally depends on its management of and success with fresh embryo transfer
— id: 106593, year: 2009, vol: 19 Suppl 3, page: 9, 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,

Early serum interleukin-8 evaluation may prove useful in localizing abnormally implanted human gestations after in vitro fertilization
Morelli, Sara S; Keegan, Debbra A; Krey, Lewis C; Katz, Joseph; Liu, Mengling; Noyes, Nicole
2008 Dec;90(6):2068-2072, Fertility & sterility
OBJECTIVE: To determine whether early measurement of the serum cytokines interleukin-2 receptor (IL-2R), IL-6, and IL-8 along with human chorionic gonadotropin (hCG) and progesterone (P(4)) can differentiate an ectopic from an intrauterine gestation. DESIGN: Retrospective analysis. SETTING: University-based fertility center. PATIENT(S): 75 women who underwent treatment with in vitro fertilization (IVF) and subsequently had an ectopic gestation (n = 15), spontaneous abortion (SAB) (n = 30), or term delivery (TD) (n = 30). INTERVENTION(S): Serum samples were obtained 14 (day 28) and 21 (day 35) days after oocyte retrieval. MAIN OUTCOME MEASURE(S): Serum concentrations of IL-2R, IL-6, IL-8, P(4), and hCG. RESULT(S): Median hCG readings on day 28 and day 35 were statistically significantly lower in the ectopic gestation group than in those with spontaneous abortion or term delivery. On day 28, median IL-8 levels were lower in the ectopic gestation group when compared with all intrauterine gestations combined. No statistically significant differences in IL-2R or IL-6 levels were noted between groups. Despite P(4) supplementation, median day-35 P(4) levels were lower in ectopic gestation than in the spontaneous abortion and term delivery cycles. CONCLUSION(S): In the setting of a rise or plateau in hCG levels, low day-28 IL-8 and day-35 P(4) levels suggested an extrauterine implantation. This assay combination may facilitate earlier diagnosis of an ectopic gestation when pregnancy location is unclear
— id: 90885, year: 2008, vol: 90, page: 2068, 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,

Depot leuprolide acetate does not adversely affect oocyte donor stimulation
Flisser E; Levine BA; Krey LC; Licciardi F
2007 ;1:18-18, Probe: the publication of research on biomedical endeavors
— id: 75315, year: 2007, vol: 1, page: 18, 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,

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,

Increased risk of pregnancy-induced hypertension in young recipients of donated oocytes
Keegan, Debbra A; Krey, Lewis C; Chang, Hung-Chi; Noyes, Nicole
2007 Apr;87(4):776-781, Fertility & sterility
OBJECTIVE: To assess the rates of select obstetric outcomes in oocyte donation (OD) recipients aged <35 years and >/=40 years and compare them to similarly aged IVF patients. DESIGN: Retrospective anonymous questionnaire study. SETTING: University-based IVF center. PATIENT(S): Live-birth outcome was experienced by 199 OD recipients and 488 IVF patients <35 or >/=40 years. INTERVENTION(S): The OD or IVF cycles. MAIN OUTCOME MEASURE(S): Response rate, pregnancy outcome, and complications. RESULT(S): Response rate was 60%. The OD recipients had significantly higher rates of pregnancy-induced hypertension (PIH) than their IVF counterparts. The OD <35 years had the highest rate (42%), followed by OD >/=40 years (26%), IVF >/=40 years (14%), and IVF patients <35 years (12%). In twin pregnancies, the rates of PIH remained higher in the OD groups: OD <35 years (56%), OD >/=40 years (36%), IVF >/=40 years (25%), and IVF <35 years (22%). Twin pregnancy rate was lowest in IVF patients >/=40 years (19%) and a lower preterm delivery rate (16%) reflected this difference. The cesarean section rates were 50% for singleton and 78% for twin deliveries in the OD patients <35 year; in OD patients >/=40 years, the rates were 75% and 84%, respectively. CONCLUSION(S): The OD recipients are at higher risk for untoward obstetric outcomes than their IVF counterparts. Young OD recipients reported the highest rate of PIH, warranting further investigation into an association between early loss of ovarian function and PIH
— id: 71128, year: 2007, vol: 87, page: 776, stat: Journal Article,

Monozygotic twinning: A 5 year experience in a large IVF center doing blastocyst transfers on day 5
Knopman, JM; Lee, J; Krey, LC; Fino, E; Adler, AA; Noyes, N
2007 SEP ;88(3):S315-S315, Fertility & sterility
— id: 74429, year: 2007, vol: 88, page: S315, 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,

Human papilloma virus (HPV) and abnormal Pap smears at an infertility clinic: Prevalence and association with fertility treatment outcome.
Kulp, JL; Krey, LC; Kwiatkowski, A; Licciardi, FL; Noyes, NL
2007 SEP ;88(3):S267-S268, Fertility & sterility
— id: 74425, year: 2007, vol: 88, page: S267, 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,

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,

Ovarian stimulation in 2007: The evolving role of GNRH analogues at a large, university-based fertility center
Reh, AE; Krey, L; Talebian, S; Licciardi, F; Noyes, N
2007 SEP ;88(3):S97-S98, Fertility & sterility
— id: 74417, year: 2007, vol: 88, page: S97, stat: Journal Article,

Oral contraceptive (OC) pre-treatment in GnRH antagonist (ANT) cycles is a reasonable option to control the timing of IVF cycles
Talebian, S; Krey, LC; Reh, A; Ganguly, N; Liu, M; Noyes, N
2007 SEP ;88(3):S291-S291, Fertility & sterility
— id: 74426, year: 2007, vol: 88, page: S291, 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,

Depot leuprolide acetate does not adversely affect oocyte donor stimulation
Flisser, E; Levine, BA; Krey, LC; Licciardi, F
2006 SEP ;86(1-2):S409-S409, Fertility & sterility
— id: 70640, year: 2006, vol: 86, page: S409, stat: Journal Article,

The myth of the "clutch" donor
Flisser, E; Licciardi, F; Krey, LC
2006 SEP ;86(1-2):S183-S184, Fertility & sterility
— id: 70629, year: 2006, vol: 86, page: S183, 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,

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,

Gestational age at delivery is not affected by controlled ovarian hyperstimulation: A comparison of IVF and donor EGG pregnancies
Salas, J; Talebian, S; Krey, LC; Licciardi, F
2006 SEP ;86(1-2):S186-S187, Fertility & sterility
— id: 70630, year: 2006, vol: 86, page: S186, 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,

Evaluating the role of exogenous luteinizing hormone in poor responders undergoing in vitro fertilization with gonadotropin-releasing hormone antagonists
Chung, Karine; Krey, Lewis; Katz, Joseph; Noyes, Nicole
2005 Aug;84(2):313-318, Fertility & sterility
OBJECTIVE: To evaluate the importance of exogenous LH in poor responders undergoing IVF with GnRH antagonists. DESIGN: Retrospective cohort study. SETTING: University-based IVF center. PATIENT(S): All patients with a history of poor response to ovarian stimulation undergoing IVF with GnRH antagonists between September 2000 and August 2001. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates. RESULT(S): Two hundred forty GnRH-antagonist cycles were initiated in poor responders. One hundred fifty-three progressed to oocyte retrieval. Seventy-five patients received recombinant FSH (Rec) for ovarian stimulation, and 66 received hMG in combination with Rec. In patients aged <40 years, there were no significant differences in amount and duration of treatment, number of oocytes retrieved, and number of embryos between treatment groups. In patients aged > or =40 years, significantly fewer oocytes were retrieved in groups who received exogenous LH in their stimulation, resulting in significantly fewer fertilized embryos. Implantation and clinical pregnancy rates did not differ by treatment group. CONCLUSION(S): In poor responders undergoing IVF with GnRH antagonists, outcomes are comparable whether stimulation is achieved in the presence or absence of supplemental LH. Exogenous LH does not appear to be necessary to achieve pregnancy in these challenging patients and may be detrimental to older patients with a history of poor response
— id: 90886, year: 2005, vol: 84, page: 313, 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,

Diminishing returns of increasing gonadotropin dosage in subsequent in vitro fertilization (IVF) cycles?
Flisser, E; Krey, LC; Berkeley, AS
2005 SEP ;84(11):S325-S326, Fertility & sterility
— id: 59571, year: 2005, vol: 84, page: S325, stat: Journal Article,

Day 1 intracytoplasmic sperm injection - Five years of "rescued" oocytes
Flisser, E; Krey, LC; Noyes, N
2005 SEP ;84(11):S372-S372, Fertility & sterility
— id: 59573, year: 2005, vol: 84, page: S372, 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,

Extended embryo culture and natural fertilization do not protect against chromosomal errors: Aneuploidy rates in spontaneous abortions (SAB) are similar for day 3 (D3) and day 5 (D5) embryo transfers and are unrelated to intracytoplasmic sperm injection (ICSI)
Keegan, DA; Krey, LC; Fritz, R; Noyes, N
2005 SEP ;84(11):S282-S283, Fertility & sterility
— id: 59567, year: 2005, vol: 84, page: S282, stat: Journal Article,

Younger (< 35 years) donor egg recipients are at high risk for pregnancy-induced hypertension (PIH): A link between premature ovarian failure and PIH?
Keegan, DA; Krey, LC; Noyes, N
2005 SEP ;84(11):S120-S120, Fertility & sterility
— id: 59564, year: 2005, vol: 84, page: S120, stat: Journal Article,

The effect of media and protein supplements as well as day of embryo transfer (ET) on monozygotic twinning (MZT) rates
Lee, HL; Adler, A; Labella, P; McCaffrey, C; Licciardi, F; Krey, LC
2005 SEP ;84(11):S374-S375, Fertility & sterility
— id: 59574, year: 2005, vol: 84, page: S374, 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,

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,

Luteal serum estradiol levels during in vitro fertilization-embryo transfer (IVF-ET) cycles do not suggest a role for estradiol supplementation
Dunham, S; Krey, LC; Noyes, N
2004 SEP ;82(2):S258-S259, Fertility & sterility
— id: 48956, year: 2004, vol: 82, page: S258, stat: Journal Article,

Donor age does not impact the success of oocyte donation cycles
Flisser, E; Kump, LM; Krey, LC; Licciardi, F
2004 SEP ;82(2):S211-S211, Fertility & sterility
— id: 48952, year: 2004, vol: 82, page: S211, 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,

Single blastocyst transfer is an effective treatment option
Huang, JQ; Krey, L; Licciardi, F
2004 SEP ;82(2):S200-S200, Fertility & sterility
— id: 48951, year: 2004, vol: 82, page: S200, stat: Journal Article,

Duration of embryo culture and intracytoplasmic sperm injection (ICSI): The effect on pregnancy and miscarriage outcome
Margulies, R; Krey, L; Noyes, N
2004 SEP ;82(2):S222-S222, Fertility & sterility
— id: 48953, year: 2004, vol: 82, page: S222, stat: Journal Article,

Use of oral contraceptives with GnRH antagonists and recombinant gonadotropins in IVF cycles have no deleterious effect on pregnancy outcome
Talebian, S; Krey, LC; Noyes, N
2004 SEP ;82(2):S234-S234, Fertility & sterility
— id: 48954, year: 2004, vol: 82, page: S234, 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,

Intrafollicular profiles of reproductive hormones in IVF treatment cycles employing GnRH antagonists and agonists
Noyes, N; Yegorov, V; Katz, J; Levitz, M; Krey, L
2003 SEP ;80(11):S106-S106, Fertility & sterility
— id: 55398, year: 2003, vol: 80, page: S106, 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,

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,

The efficacy and safety of the use of sildenafil as an aid to ejaculatory failure at the time of in vitro fertilization/intracytoplasmic sperm injection
Adler, A; McCullough, A; Patel, R; Krey, LC
2002 SEP ;78(3):S261-S261, Fertility & sterility
— id: 55578, year: 2002, vol: 78, page: S261, stat: Journal Article,

Prognosis for live birth with IVF after elevated day 3 FSH
Brown, JR; Krey, L
2002 SEP ;78(3):S248-S248, Fertility & sterility
— id: 55577, year: 2002, vol: 78, page: S248, 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,

Follicular serum androgen profiles differ when GnRH agonists and antagonists are used
Chung, K; Krey, L; Ambrosini, G; Katz, J; Noyes, N
2002 SEP ;78(3):S173-S174, Fertility & sterility
— id: 55572, year: 2002, vol: 78, page: S173, stat: Journal Article,

Preimplantation genetic diagnosis (PGD), a collaborative activity of clinical genetic departments and IVF centres (vol 21, pg 1086, 2001)
Geraedts, JPM; Harper, J; Braude, P; Sermon, K; Veiga, A; Gianaroli, L; Agan, N; Munne, S; Gitlin, S; Blenow, E; De Boer, K; Hussey, N; Traeger-Synodinos, J; Lee, SH; Viville, S; Krey, L; Ray, P; Emiliani, S; Liu, YH; Vermeulen, S
2002 May;22(5):451-451, Prenatal diagnosis
— id: 28184, year: 2002, vol: 22, page: 451, 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,

Predictive value of polymorphonuclear granulocytes in relation to growth of microorganisms in semen
Lu, L; Look, C; Sanchez, XM; Lacsamana, J; Macanas, E; Krey, LC
2002 SEP ;78(3):S86-S86, Fertility & sterility
— id: 55568, year: 2002, vol: 78, page: S86, stat: Journal Article,

Prescribed medications and sperm production and function during IVF
Lu, L; Sanchez, X; Look, C; Lacsamana, J; Macanas, E; Krey, L
2002 SEP ;78(3):S232-S233, Fertility & sterility
— id: 55576, year: 2002, vol: 78, page: S232, 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,

Estradiol: Oocyte ratio an early predictor of reduced ovarian reserve
Montoya, A; Licciardi, F; Krey, L
2002 SEP ;78(3):S50-S50, Fertility & sterility
— id: 55566, year: 2002, vol: 78, page: S50, stat: Journal Article,

Do hydrosalpinges affect IVF outcome?
Noyes, N; Jones, K; Patel, A; Kwiakowski, A; Ryan, C; Krey, LC
2002 SEP ;78(3):S144-S144, Fertility & sterility
— id: 55569, year: 2002, vol: 78, page: S144, stat: Journal Article,

GnRH-antagonists: Useful adjuncts for stimulation in women with poor ovarian reserve
Noyes, N; Krey, LC
2002 SEP ;78(3):S173-S173, Fertility & sterility
— id: 55571, year: 2002, vol: 78, page: S173, stat: Journal Article,

An oral contraceptive (OCP) microdose flare leuprolide acetate (LA) stimulation protocol is useful for poor prognosis patients undergoing in vitro fertilization (IVF)
Sanghvi, A; Noyes, N; Krey, LC
2002 SEP ;78(3):S149-S149, Fertility & sterility
— id: 55570, year: 2002, vol: 78, page: S149, stat: Journal Article,

Pregnancy outcome and complications in women age 40 or older undergoing in-vitro fertilization with autologous and donor oocytes
Styne, A; Krey, LC; Kwiatkowski, A; Licciardi, F; Noyes, N
2002 SEP ;78(3):S9-S10, Fertility & sterility
— id: 55564, year: 2002, vol: 78, page: S9, 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,

Preimplantation genetic diagnosis (PGD), a collaborative activity of clinical genetic departments and IVF centres
Geraedts, JPM; Harper, J; Braude, P; Sermon, K; Veiga, A; Gianaroli, L; Agan, N; Munne, S; Gitlin, S; Blenow, E; de Boer, K; Hussey, N; Kanavakis, E; Lee, SH; Viville, S; Krey, L; Ray, P; Emiliani, S; Liu, YH; Vermeulen, S
2001 DEC ;21(12):1086-1092, Prenatal diagnosis
Preimplantation genetic diagnosis (PGD) requires the combined efforts of geneticists and workers in (lie field of reproductive medicine. This was studied on the basis of a questionnaire, sent to 35 members of the PGD Consortium of the European Society of Human Reproduction and Embryology (ESHRE). A reply was obtained from 20 centres. They represent the majority of activities in the field of PGD in the world. It is obvious that many of the activities (in vitro fertilisation, embryo culture and biopsy) take place in IVF units while others (counselling and diagnosis) are the responsibility of genetic diagnostic centres. The distances between both units vary considerably. In all but one centre sex determination is offered. Aneuploidy screening is offered in 13 out of 20 centres. PGD of translocations and other structural chromosome abnormalities is offered in all but one centre. The number of monogenic diseases offered varies considerably. In comparison to prenatal diagnosis PGD is more expensive. The majority of these costs are due to the IVF or ICSI procedure. The charges for PGD vary between about 600 euro and 4000 euro. In 16 out of 20 centres the parents to be must sign an informed consent form.
— id: 55355, year: 2001, vol: 21, page: 1086, 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,

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,

Crinone (R) vaginal gel vs. intramuscular progesterone for luteal support in oocyte donation cycles: clinical and endocrinologic differences
Licciardi, FL; Sanghvi, A; Lawson, G; Krey, LC
2001 SEP ;76(3):S168-S169, Fertility & sterility
— id: 54923, year: 2001, vol: 76, page: S168, 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,

Normal live birth produced from preovulatory mouse oocytes reconstructed by germinal vesicle transfer
Liu, H; Krey, LC; Zhang, J; Grifo, LA
2001 SEP ;76(3):S56-S56, Fertility & sterility
— id: 54916, year: 2001, vol: 76, page: S56, stat: Journal Article,

A comparison of in vitro fertilization (IVF) success rates in non obstructive azoospermia (NOA) with fresh (FRTB) and cryopreserved (FZTB) testicular tissue
McCullough, AR; Bar Chama, N; Adler, A; Gans, W; Krey, L
2001 SEP ;76(3):S247-S247, Fertility & sterility
— id: 54927, year: 2001, vol: 76, page: S247, stat: Journal Article,

A comparison of in vitro fertilization (IVF) pregnancy and delivery rates in acquired (AA) and congenital (CA) obstructive azoospermia
McCullough, AR; Bar Chama, N; Gans, W; Adler, A; Clarke, M; Krey, L
2001 SEP ;76(3):S242-S242, Fertility & sterility
— id: 54926, year: 2001, vol: 76, page: S242, stat: Journal Article,

Are frozen epididymal sperm (FZEP) comparable to fresh testicular (FRTB) sperm in men with obstructive azoospermia (OA)?
McCullough, AR; Chama, NB; Adler, A; Gans, W; Krey, L
2001 SEP ;76(3):S140-S141, Fertility & sterility
— id: 54921, year: 2001, vol: 76, page: S140, 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,

GnRH-antagonists (GN-Ant): useful adjuncts for stimulation in women with poor ovarian reserve
Noyes, N; Pan, L; Berkeley, AS; Krey, LC
2001 SEP ;76(3):S175-S175, Fertility & sterility
— id: 54924, year: 2001, vol: 76, page: S175, stat: Journal Article,

Blastocyst transfer: does early embryonic evaluation and/or ultimate embryo selection affect prognosis?
Song, E; McCaffrey, C; Krey, LC; Grimes, D; Noyes, N
2001 SEP ;76(3):S87-S87, Fertility & sterility
— id: 54918, year: 2001, vol: 76, page: S87, stat: Journal Article,

Multinucleated embryos in IVF: associated clinical factors and pregnancy outcome
Styne, AJ; Chi, L; Krey, LC; Adler, A; Noyes, N
2001 SEP ;76(3):S144-S145, Fertility & sterility
— id: 54922, year: 2001, vol: 76, page: S144, 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,

Enhancement or initiation of testicular sperm motility by in vitro culture of testicular tissue
Angelopoulos T; Adler A; Krey L; Licciardi F; Noyes N; McCullough A
1999 Feb;71(2):240-243, Fertility & sterility
OBJECTIVE: To describe different techniques of testicular tissue culture and their effect on sperm motility, mainly in cases of totally immotile spermatozoa, and to compare the effect of in vitro culture with that of motility stimulants. DESIGN: Prospective study. SETTING: University teaching hospital. PATIENT(S): Ten patients undergoing testicular biopsy for diagnostic purposes or for intracytoplasmic sperm injection. INTERVENTION(S): Dissected testicular biopsy samples and tissue blocks were cultured at 37 degrees C for up to 96 hours. Immediately after dissection, immotile testicular spermatozoa were incubated for 30 minutes in pentoxifylline and 2-deoxyadenosine. MAIN OUTCOME MEASURE(S): Sperm motility and vitality. RESULT(S): Overall, dissected samples showed improved sperm motility, which peaked within 48 hours of culture. Unlike motility, vitality declined linearly, from 56.3%+/-19% at initial assessment to 18.8%+/-11% at 96 hours. Five samples had initially immotile spermatozoa, of which four acquired motility at 48 hours. In vitro culture showed results comparable with those of incubation with pentoxifylline and 2-deoxyadenosine. Culture of tissue blocks did not improve motility or vitality compared with dissected tissue. CONCLUSION(S): The motility of testicular spermatozoa was enhanced or initiated after in vitro culture. Testicular biopsy culture may be an alternative to the use of motility stimulants to obtain motile spermatozoa for intracytoplasmic sperm injection, particularly when oocytes are not immediately available
— id: 57018, year: 1999, vol: 71, page: 240, 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,

The antiviral agents, MAP30 and GAP31, are not toxic to human spermatozoa and may be useful in preventing the sexual transmission of human immunodeficiency virus type 1
Schreiber CA; Wan L; Sun Y; Lu L; Krey LC; Lee-Huang S
1999 Oct;72(4):686-690, Fertility & sterility
OBJECTIVE: To investigate the effects of two virucidal compounds, MAP30 (Momordica anti-human immunodeficiency virus [HIV] protein; molecular weight, 30 kd) and GAP31 (Gelonium anti-HIV protein; molecular weight, 31 kd), obtained from Momordica charantia and Gelonium multiflorum, respectively, on the motility and vitality of human spermatocytes. DESIGN: Prospective, controlled study. SETTING: New York University School of Medicine. PATIENT(S): Ten healthy men undergoing evaluation for infertility provided 10 semen specimens. INTERVENTION(S): Human sperm were treated with the anti-HIV agents, MAP30 and GAP3 1. Nonoxynol-9, a commonly used spermicide, and phosphate-buffered saline were used as the positive and negative controls, respectively. MAIN OUTCOME MEASURE(S): The motility and vitality of human spermatocytes treated with MAP30 and GAP31 at doses that inhibit HIV-1 and herpes simplex virus. RESULT(S): MAP30 and GAP31 did not inhibit the motility or vitality of human sperm cells over a dose range of 100-0.1 microg/mL, whereas nonoxynol-9 demonstrated spermicidal action on all 10 samples over the same dose range. CONCLUSION(S): The antiviral agents, MAP30 and GAP31, were not toxic to human sperm cells at the doses at which they inhibit HIV-1 and herpes simplex virus. They had no effect on the motility of spermatozoa, even at a dose of 1,000 times the maximum effective concentration. These results indicate that MAP30 and GAP31 may be useful as nonspermicidal protection against sexually transmitted diseases
— id: 11946, year: 1999, vol: 72, page: 686, 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,

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,

Effects of photoperiod on brain corticosteroid receptors and the stress response in the golden hamster (Mesocricetus auratus)
Ronchi E; Spencer RL; Krey LC; McEwen BS
1998 Jan 12;780(2):348-351, Brain research
Following exposure to short daylengths, in golden hamsters, changes in basal adrenal glucocorticoid secretion are associated with a significant increase in Type I receptor binding, and are preceded by alterations in the stress-induced release of glucocorticoids, which is one of the major modes of operation of the hypothalamo-pituitary-adrenocortical axis (HPA). These results lend support to the hypothesis that corticosteroid receptors, and in particular the Type I receptor subtype, play a central role in the regulation of circadian and circannual rhythms of the HPA
— id: 55819, year: 1998, vol: 780, page: 348, 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,

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,

Alterations in endometrial stromal cell tissue factor protein and messenger ribonucleic acid expression in patients experiencing abnormal uterine bleeding while using Norplant-2 contraception
Runic R; Schatz F; Krey L; Demopoulos R; Thung S; Wan L; Lockwood CJ
1997 Jun;82(6):1983-1988, Journal of clinical endocrinology & metabolism
A high incidence of irregular uterine bleeding is the primary patient complaint limiting the utility of long term, progestin-only contraceptive agents such as Norplant. The onset of hemorrhage requires both inadequate hemostasis and impaired vascular integrity. Thus, we first tested whether Norplant-associated endometrial bleeding was accompanied by altered expression of perivascular stromal cell tissue factor (TF), the primary initiator of hemostasis. Norplant effects on TF messenger ribonucleic acid (mRNA) and protein expression by endometrial stromal cells were assessed by in situ hybridization and immunohistochemical examination of endometrial biopsies obtained from normally cycling control women (n = 14) and from patients experiencing Norplant-induced abnormal uterine bleeding (n = 24). TF mRNA and protein expression was increased 150% in secretory vs. proliferative phase endometrial specimens. By contrast, endometrial TF mRNA and protein levels were reduced during 1-6 months of Norplant treatment by about 2-fold (P < 0.05 for protein) compared to the values for control secretory phase specimens. These changes were consistent with observations that patients on Norplant begin to bleed during this interval. Further reductions of TF mRNA and protein levels to 2- and 3-fold of those in secretory phase control specimens were observed in endometria obtained after 6-12 months of Norplant therapy (P < 0.05 and P < 0.01, respectively). A modest rebound in TF mRNA and protein expression was observed after 12 months of Norplant therapy, which occurred commensurate with reduced patient complaints of abnormal uterine bleeding. Pathologically enlarged venous sinusoids were ubiquitous in endometrial specimens obtained after Norplant therapy. The combination of fragile blood vessels and reduced TF expression may account for bleeding in patients receiving Norplant therapy
— id: 12314, year: 1997, vol: 82, page: 1983, stat: Journal Article,

The embryo versus endometrium controversy revisited as it relates to predicting pregnancy outcome in in-vitro fertilization-embryo transfer cycles
Schwartz LB; Chiu AS; Courtney M; Krey L; Schmidt-Sarosi C
1997 Jan;12(1):45-50, Human reproduction
To evaluate embryonic and endometrial factors for their value in predicting pregnancy outcome in in-vitro fertilization (IVF) and embryo transfer, a retrospective data collection and prospective uterine artery colour Doppler imaging study was performed in a university-based IVF-embryo transfer programme setting. A total of 210 patients were included and grouped as follows: (I) IVF with controlled ovarian stimulation (214 cycles); (II) frozen-thaw cycle of autologous embryos (30 cycles); (III) oocyte donation, no cryopreservation (12 cycles); (IV) frozen-thaw cycle with embryos from donated oocytes (10 cycles). Embryo quality was significantly better in pregnant than non-pregnant cycles (group I, P = 0.0104; groups II-IV, P = 0.0418). The endometrial echo was significantly thicker in pregnant versus non-pregnant patients in group I (P = 0.0059), but not in groups II-IV (P = 0.741). Past uterine surgery or abnormalities had no effect on pregnancy outcome. There were no significant differences in mean uterine artery resistance index or peak systolic velocity in pregnant versus non-pregnant patients in groups II-IV. Thus, embryo quality is the most reliable predictor of pregnancy outcome. Endometrial measurements were significantly thicker in subsequently pregnant patients only in group I, where the endometrium reflects the hormonal environment. Doppler parameters were not useful in predicting pregnancy outcome
— id: 12418, year: 1997, vol: 12, page: 45, stat: Journal Article,

Alterations in steroid hormone receptors in the tamoxifen-treated endometrium
Schwartz LB; Krey L; Demopoulos R; Goldstein SR; Nachtigall LE; Mittal K
1997 Jan;176(1 Pt 1):129-137, American journal of obstetrics & gynecology
OBJECTIVE: Our purpose was to evaluate whether tamoxifen has estrogenic endometrial effects as defined by histologic study or alterations in steroid hormone receptor expression. STUDY DESIGN: Nineteen postmenopausal tamoxifen-treated breast cancer patients who also had endometrial sampling were identified from files in the Department of Obstetrics and Gynecology. To examine the subgroup of 15 polyps, age-matched, non-hormonally treated patients with polyps (n = 8) or atrophic endometria (n = 5) served as comparison groups. Proliferative (n = 3) and secretory (n = 5) endometria served as procedural controls. Immunohistochemical studies for steroid receptors (estrogen, progesterone) were performed. RESULTS: Glandular cell progesterone receptor was significantly increased and stromal cell estrogen receptor was significantly decreased in tamoxifen-treated versus atrophic endometria. Progesterone receptor staining was not significantly different in tamoxifen-treated versus control polyps, although staining was high in both groups. Stromal cell estrogen receptor staining was significantly reduced in tamoxifen-treated versus control polyps, although there were no histologic differences. Reduced stromal cell estrogen receptor and increased glandular cell progesterone receptor staining was found in all tamoxifen-treated endometria regardless of the diagnosis. CONCLUSION: The tamoxifen-associated changes in endometrial steroid receptors support an estrogenic effect that is independent of histologic diagnosis and duration of use. This may contribute to the pathogenesis of tamoxifen-associated polyps and carcinomas
— id: 7784, year: 1997, vol: 176, page: 129, stat: Journal Article,

Thyroid hormone and estrogen interact to regulate behavior
Dellovade, TL; Zhu, YS; Krey, L; Pfaff, DW
1996 OCT 29 ;93(22):12581-12586, Proceedings of the National Academy of Sciences of the United States of America
Environmental perturbations that increase plasma thyroid hormone (T-3) concentrations also profoundly affect female reproductive behavior and physiology, We explored whether these effects were mediated by interactions between T-3 receptor (TR) and estrogen receptor (ER). This hypothesis was of interest because the half-site of a consensus T-3 response element DNA sequence is identical to an ER response element (ERE), and TRs bind to a consensus ERE. Molecular data presented in the accompanying paper [Zhu, Y.-S., Yen, P. M,, Chin, W, W, & Pfaff, D, W, (1996) Proc, Natl. Acad, Sci. USA 93, 12587-12592] demonstrate that TRs and ERs are both present in rat hypothalamic nuclear extracts and that both can bind to the promoter the hypothalamic gene preproenkephalin and that interations between liganded TRs and ERs affect preproenkephalin transcription, In this paper, we show that molecular interactions between TRs and ERs are sufficient to mediate environmental effects on estrogen-controlled reproductive behavior, Ovariectomized (OVX) rats treated with high doses of T-3 showed significantly lower levels of lordosis behavior in response to estradiol benzoate (EB) compared with OVX females treated with EB alone, Conversely, thyroidectomized/OVX females treated with EB showed significantly greater levels of lordosis behavior compared with OVX females treated with EB, showing the effect of endogenous T-3. Thyroid hormone interference with EB-induced behavior could not be explained by a reduction in plasma E(2) concentrations or by a general reduction in responsiveness of EB-sensitive tissues. Moreover, numbers of hypothalamic EB-immunoreactive cells increased dramatically following T-3 treatment. These data suggest that T-3 may reduce EB-dependent sexual behavior through interactions between TR and ER in the nuclei of behaviorally relevant hypothalamic neurons, envisioning for the first time a functional consequence of interactions between two nuclear hormone receptors in brain. These results also open up the possibility of molecular interactions on DNA encoding environmental signals, a new field for the study of neuronal integration
— id: 52750, year: 1996, vol: 93, page: 12581, 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,

Preliminary observations on the use of midodrine in treating orthostatic hypotension in familial dysautonomia
Axelrod FB; Krey L; Glickstein JS; Allison JW; Friedman D
1995 Oct 5;55(1-2):29-35, Journal of the autonomic nervous system
Midodrine, a peripheral alpha-adrenergic agonist, was evaluated in 7 female and 2 male patients with familial dysautonomia (FD), a disorder characterized by decreased sympathetic innervation. Prior to and after three months of midodrine treatment, each patient's response to postural change was assessed by arteriosonde readings of blood pressure and heart rate, corrected QT-interval measurements, Doppler evaluation of renal blood flow and circulating atrial natriuretic peptide (ANP) levels. The initial midodrine dose (2.5 mg three times daily) was raised until subjective symptoms improved. Doses were reduced if patients felt jittery or developed erect hypertension (systolic > 180 mmHg or diastolic > 110 mmHg). Midodrine, at an average dose of 0.25 mg/kg per day, improved subjective symptoms in all patients. With treatment, magnitude of blood pressure responses was variable. Although mean erect blood pressure did not increase significantly for the aggregate, it did increase in six of nine patients. In addition, the QTc interval normalized and erect renal perfusion improved. Changes in supine mean blood pressure and supine circulating ANP correlated directly. We judge midodrine to be useful in management of orthostatic hypotension in patients with familial dysautonomia
— id: 6803, year: 1995, vol: 55, page: 29, stat: Journal Article,

Atrial natriuretic peptide response to postural change and medication in familial dysautonomia
Axelrod FB; Krey L; Glickstein JS; Friedman D; Weider J; Metakis LJ; Porges VM; Mineo M; Notterman D
1994 Dec;4(6):311-318, Clinical autonomic research
Circulating atrial natriuretic peptide (ANP) was assayed before and after postural change and exercise in 54 patients with familial dysautonomia (FD) and 20 controls. ANP levels were compared with blood pressure, heart rate, plasma catecholamines and parameters of renal function. Compared with controls supine FD subjects had elevated blood pressures, heart rates and ANP levels (39 +/- 4 pg/ml vs. 23 +/- 3 pg/ml, p < 0.01). With the erect posture and exercise in FD subjects, blood pressure fell below control values, with ANP lowered. In FD subjects, blood pressure was correlated with ANP levels when supine and when erect and with heart rate post exercise. In controls, ANP levels did not correlate with other parameters. In FD patients on metoclopramide, supine and erect blood pressure and ANP levels were higher. FD subjects treated with fludrocortisone, had elevated supine and erect noradrenaline (p < 0.05 and p = 0.06); and those on diazepam had lower erect and post exercise noradrenaline (p < 0.05), but ANP levels were similar. In conclusion, sympathetic denervation may increase FD patients' responsiveness to other regulators of cardiovascular integrity, such as ANP. In addition, circulating ANP and catecholamines in FD subjects appear to be influenced by commonly used medications, such as metoclopramide
— id: 6572, year: 1994, vol: 4, page: 311, stat: Journal Article,

ACTIONS OF GNRH AGONISTS ON GONADOTROPE PHYSIOLOGY
KREY, LC
1993 MAY ;11(2):105-111, Seminars in reproductive endocrinology
— id: 52167, year: 1993, vol: 11, page: 105, stat: Journal Article,

Immunocytochemical study of GnRH and GnRH-associated peptide in male Syrian hamsters as a function of photoperiod and gonadal alterations
Ronchi E; Aoki C; Krey LC; Pfaff DW
1992 Feb;55(2):134-145, Neuroendocrinology
Hypothalamic luteinizing-hormone-releasing hormone (GnRH) and gonadotropin-releasing-hormone-associated peptide (GAP) biosynthesis and storage were estimated by immunocytochemistry in male golden hamsters maintained in different photoperiods. Intact or castrated male hamsters with subcutaneously inserted testosterone implants were exposed to long-day (14:10) or short-day photoperiods (10:14) for 4-8 weeks. Exposure to short photoperiod for 4 weeks, an interval characterized by a suppression of gonadotropin secretion but not gonadal regression, was associated with an increase in the number of GnRH- and GAP-immunoreactive cells in the diagonal band of Broca/medial septum. Furthermore, morphometric analysis revealed that these animals displayed significantly more GnRH but not GAP immunoreactivity in the median eminence as opposed to hamsters exposed to long-day photoperiods. In additional studies, gonadally regressed hamsters exposed to short day lengths for 8 weeks had equal numbers of GnRH cells as did the long-day controls. These patterns suggest that reproductive quiescence in golden hamsters is not the result of depletions of neuronal GnRH stores available for secretion
— id: 55820, year: 1992, vol: 55, page: 134, stat: Journal Article,

Steady state analysis of hypothalamic GnRH mRNA levels in male Syrian hamsters: influences of photoperiod and androgen
Ronchi E; Krey LC; Pfaff DW
1992 Feb;55(2):146-155, Neuroendocrinology
An in situ hybridization assay, utilizing a free floating technique was used to estimate the steady state levels of hypothalamic luteinizing hormone-releasing hormone (GnRH) mRNA levels in the brains of male golden hamsters maintained in different photoperiods. In situ histochemistry was performed using a 32P-labelled 66-nucleotide long oligomer complementary to the sequence of the human GnRH mRNA coding region. The oligonucleotide hybridized specifically to mRNA encoding the GnRH precursor as suggested by the distribution of labelled neurons and as shown by an RNAse protection assay on septal and preoptic-hypothalamic mRNA from gonadally regressed hamsters. To test the hypothesis that short-day photoperiods reduce GnRH synthesis, intact male hamsters or castrated males bearing subcutaneously inserted testosterone implants were exposed to long-day (14 h light:10 h dark) or short-day (10 h light 14 h dark) photoperiods for 4 weeks. Exposure to short day lengths never caused a decrease in GnRH expressing neurons and actually was associated with an increase in the number of radiolabelled cells specifically in the diagonal band of Broca/medial septum in the gonadally intact group. The mean number of grains per labelled cell for the short day animals similarly was not reduced from that seen in long day animals. The results are consistent with previous studies on photoperiod and GnRH content in the same brain regions and support the notion that the suppression of the synthesis of GnRH does not accompany the low levels of LH secretion observed during the early stages of reproductive quiescence in this species
— id: 55821, year: 1992, vol: 55, page: 146, stat: Journal Article,

Testosterone processing by pituitary cells in culture: an examination of the role of 5 alpha-reduction in androgen action on the gonadotroph
Kamel F; Krey LC
1991 Jan;56(1):22-29, Steroids
Dispersed rat pituitary cells were exposed to [1,2,6,7-3H]testosterone ([3H]T, 10(-8) M) to assess the role of 5 alpha-reduction in T regulation of gonadotroph secretion. After 4 to 48 hours of exposure, [3H]T metabolites isolated by thin-layer chromatography were characterized in medium and cell homogenates as well as bound to androgen receptors salt-extracted from purified nuclear pellets. Receptor-bound 5 alpha-[3H]dihydrotestosterone ([3H]DHT)/total [3H]androgens rose progressively from 16% at 4 hours to more than 50% at 48 hours. Coincubation with 4-MA (10- to 1,000-fold molar excess) or testosterone-17 beta-carboxylic acid (TCA; 1,000-fold excess) reduced receptor-bound [3H]DHT/[3H]androgen to less than 10% and 20%, respectively, but elevated [3H]T-receptor levels. Despite inhibiting 5 alpha-reductase activity, TCA and 4-MA had no effect on T suppression of gonadotropin-releasing hormone-stimulated luteinizing hormone secretion or T enhancement of total (cell + secreted) follicle-stimulating hormone levels. The results suggest that 5 alpha-reduction to DHT is not essential for the expression of the direct influences of T on gonadotropin synthesis and secretion in rat gonadotrophs
— id: 55823, year: 1991, vol: 56, page: 22, stat: Journal Article,

Estradiol processing by pituitary cells in culture: an examination of the influences of various exposures to progesterone
Krey LC; Kamel F
1990 ;47(14):1235-1241, Life sciences
Dispersed, estradiol (E2)-treated pituitary cells were used to examine the cellular mechanisms underlying progesterone (P) suppression of GnRH-stimulated LH and FSH secretion. When cells were exposed to 10(-9) M E2 for 48 h prior to GnRH challenge, P (10(-7) M) treatment for the last 24 h suppressed gonadotroph responsiveness to GnRH for both LH and FSH secretion (gonadotropin released/intracellular stores of gonadotropin available for release). To determine if P acts by blocking E2 processing and/or uptake, we exposed cells to 2,4,6,7-3H-E2 +/- P and monitored the level and distribution of 3H-estrogens bound to estrogen receptors salt-extracted from nuclear pellets purified by sucrose density centrifugation. At 1, 4 and 24 h, P had no effect on the level of 3H-estrogen+receptor complexes or on the distribution of receptor-bound 3H-E2, 3H-estrone and 3H-estriol. The results indicate that chronic influences of P to suppress the responsiveness of E2-treated gonadotrophs to GnRH cannot be explained by alterations in estrogen receptor occupation as is the case in reproductive tract tissues
— id: 55827, year: 1990, vol: 47, page: 1235, stat: Journal Article,

Progesterone modulation of gonadotropin secretion by dispersed rat pituitary cells in culture. I. Basal and gonadotropin-releasing hormone-stimulated luteinizing hormone release
Krey LC; Kamel F
1990 Jan 22;68(2-3):85-94, Molecular & cellular endocrinology
Dispersed, estradiol-treated, rat pituitary cells were cultured to characterize the influences of a physiologic concentration of progesterone (P, 10(-7) M) on gonadotroph responsiveness to gonadotropin-releasing hormone (GnRH). Acute (less than 6 h) P treatment enhanced and chronic (greater than 12 h) treatment suppressed both basal and GnRH-stimulated luteinizing hormone (LH) release. This modulation took place without any change in intracellular LH stores, indicating that the secretory changes are not attributable to changes in LH synthesis, and were not accompanied by similar alterations in basal or thyrotropin-releasing hormone-stimulated prolactin secretion. Moreover, the timing of these responses was fixed since a 10-fold lower P concentration produced only smaller and briefer alterations in LH release. Analyses of the temporal characteristics of effective P stimuli indicated that a brief 6 h exposure to P inhibited GnRH-stimulated LH secretion 18 h later. In contrast, P's acute actions rapidly dissipated following removal of the steroid from the culture medium. Finally, P-induced enhancement and suppression of GnRH-stimulated LH release could be blocked by appropriately timed treatments with protein synthesis inhibitors. Our findings are consistent with the hypothesis that P influences gonadotroph secretory function via the production of specific proteins
— id: 55826, year: 1990, vol: 68, page: 85, stat: Journal Article,

Progesterone modulation of gonadotropin secretion by dispersed rat pituitary cells in culture. III. A23187, cAMP, phorbol ester and DiC8-stimulated luteinizing hormone release
Krey LC; Kamel F
1990 Mar 26;70(1):21-29, Molecular & cellular endocrinology
Dispersed estradiol-treated rat pituitary cells were used to characterize progesterone (P) modulation of luteinizing hormone (LH) secretion in response to a variety of pharmacologic secretagogues which influence cell biochemistry. Acute (less than 3 h) and chronic (24 h) exposures to P prior to secretagogue challenge respectively enhanced and inhibited Ca2+ ionophore (A23187)-stimulated and gonadotropin-releasing hormone (GnRH)-stimulated LH release in similar quantitative fashion without any effect on concurrent prolactin release. Similar responses were also noted with cholera toxin-stimulated secretion. However, when protein kinase C activators such as phorbol esters and dioctanoylglycerol were used to trigger LH release, chronic exposure to P did not inhibit, but rather enhanced, LH release. Again, P had no effect on prolactin release. 'Washout' studies indicated that chronic treatments with P would suppress LH secretion stimulated by these compounds, but only when the steroid was cleared from the cells 4 h beforehand. These studies provide further evidence that P specifically modulates gonadotroph secretory function via mechanisms which bypass GnRH receptors. Moreover, they suggest that P exerts many different actions within the gonadotroph and question the role of protein kinase C in GnRH action
— id: 55824, year: 1990, vol: 70, page: 21, stat: Journal Article,

Progesterone modulation of gonadotropin secretion by dispersed rat pituitary cells in culture. II. Intracellular metabolism and progestin receptors
Krey LC; Kamel F; MacLusky NJ
1990 Jan 22;68(2-3):95-103, Molecular & cellular endocrinology
Dispersed, estradiol (E2)-treated, rat pituitary cell cultures were used to examine the intracellular processing of progesterone (P) associated with its modulation of gonadotropin-releasing hormone (GnRH)-stimulated luteinizing hormone (LH) secretion. Enhancement and suppression of LH release was only observed with acute and chronic exposures to P or other naturally occurring and synthetic progestins avidly bound by pituitary progestin receptors; such responses were inhibited by cotreatment with the antiprogestin RU486 but not with the antiandrogen flutamide, illustrating the importance of the P + receptor interactions. However, cotreatment with a 100-fold molar excess of the 5 alpha-reductase inhibitor 17 beta-N,N-diethyl-carbamoyl-4-methyl-4-aza-5 alpha-androstan-3-one (4-MA) had no effect on the expression of P's modulatory actions. Additional studies using different E2 pretreatments revealed that P enhanced LH release when progestin receptor levels were elevated. Moreover, the magnitude and duration of P's influences on LH release increased in cells with higher receptor levels. However, there were several instances in which progestin receptor level and P modulation of LH release did not correlate. In several instances E2-induced progestin receptor levels stabilized at a maximal level whereas P enhancement of LH secretion continued to increase in size and duration. These findings underscore the importance of progestin receptors for P-induced modulation of LH secretion and illustrate that 5 alpha-reduction and further metabolism of P is not obligatory for the expression of these responses. In addition, our data demonstrate that the important cellular mechanisms underlying E2 priming of gonadotroph responsiveness to P entail the induction of progestin receptor levels and other as yet unidentified cellular processes
— id: 55825, year: 1990, vol: 68, page: 95, stat: Journal Article,

Effects of daylength on androgen metabolism and pulsatile luteinizing hormone secretion in male golden hamsters
Krey LC; Ronchi E; Bittman EL
1989 Nov;50(5):533-542, Neuroendocrinology
In an effort to understand the potential neuroendocrine mechanisms underlying photoperiodic control of fertility in seasonally breeding species, we monitored the intracellular processing and nuclear uptake of [1 alpha, 2 alpha-3H]testosterone (3H-T) within the brain-pituitary complex as well as the patterns of episodic luteinizing hormone (LH) secretion in male golden hamsters exposed to long day (14 h light:10 h dark) and short day (10 h light:14 h dark) photoperiods. Target tissue specific patterns of nuclear 3H-androgens and estrogens were observed in castrated, T-replaced hamsters exposed to long and short days for 7 weeks or longer. Significantly, 3H-T metabolism or receptor-mediated nuclear uptake in the hamsters in short days was not influenced in any manner that would explain their increased responsiveness to androgen feedback suppression of LH release. Comparable patterns of episodic LH secretion were observed in acutely catheterized hamsters castrated for 7 weeks prior to exposure to 8 weeks of long or short days. Similar patterns were also observed in animals maintained in long days and castrated 1 or 2 weeks prior to blood collection. However, such a pattern was not seen in acutely castrated hamsters maintained in the short-day photoperiod. The data suggest that steroid-independent mechanisms play an important role in suppressing gonadotropin release in short days in this species. However, such mechanisms appear to be most effective when the animals are or have recently been exposed to circulating androgens
— id: 55822, year: 1989, vol: 50, page: 533, stat: Journal Article,

Are catechol oestrogens obligatory mediators of oestrogen action in the central nervous system? II. Potencies of natural and synthetic oestrogens for induction of gonadotrophin release and female sexual behaviour in the rat
MacLusky, N J; Krey, L C; Parsons, B; Merriam, G R; Loriaux, D L; Pfeiffer, D G; Naftolin, F
1986 Sep;110(3):499-505, Journal of endocrinology
The role of catechol oestrogen formation in the mechanism by which circulating oestrogens facilitate gonadotrophin release and female sexual behaviour was explored in adult female rats. The effects of oestradiol-17 beta were compared with those of a group of oestrogens with either a reduced affinity for oestrogen receptors (oestradiol-17 alpha) or a reduced ability to act as substrates for catechol oestrogen formation (2-fluoro-oestradiol, 4-fluoro-oestradiol and moxestrol (11 beta-methoxy-17 alpha-ethynyloestradiol]. Rats were ovariectomized on the evening of dioestrus day 1 of the 4-day oestrous cycle and implanted s.c. 12 h later with infusion pumps containing either one of the test oestrogens or vehicle alone. Infusion rates for oestradiol-17 beta, moxestrol, 2-fluoro-oestradiol and 4-fluoro-oestradiol were adjusted to give concentrations of nuclear oestrogen receptors in the brain and pituitary gland within the range of those found in intact female rats during pro-oestrus. Oestradiol-17 alpha was infused at the same and at a tenfold higher rate than that of oestradiol-17 beta; neither of these treatments with oestradiol-17 alpha significantly increased brain or pituitary gland nuclear oestrogen receptor levels. On the day after the pump was implanted, samples of tail vein blood were withdrawn at 12.00, 14.00, 16.00 and 18.00 h for LH assay. All animals were then injected s.c. with 1 mg progesterone in propylene glycol, and tested for feminine sexual behaviour 5 h later. Oestradiol-17 beta, moxestrol, 2-fluoro-oestradiol and 4-fluoro-oestradiol all elicited pronounced LH surges and facilitated progesterone-triggered proceptive and lordosis behaviours.(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 102785, year: 1986, vol: 110, page: 499, stat: Journal Article,

Are catechol oestrogens obligatory mediators of oestrogen action in the central nervous system? I. Characterization of pharmacological probes with different receptor binding affinities and catechol oestrogen formation rates
Pfeiffer, D G; MacLusky, N J; Barnea, E; Naftolin, F; Krey, L C; Loriaux, D L; Merriam, G R
1986 Sep;110(3):489-497, Journal of endocrinology
In an attempt to define pharmacological probes with which to test the role of catechol oestrogen formation in the central nervous system, five oestrogens (oestradiol-17 beta, oestradiol-17 alpha, 4-fluoro-oestradiol, 2-fluoro-oestradiol and moxestrol (11 beta-methoxy-17 alpha-ethynyloestradiol) were studied for binding to oestrogen receptors and conversion to catechol metabolites. Binding to cytosol oestrogen receptors was measured in the hypothalamus-preoptic area-amygdala (HPA), pituitary gland and uterus of ovariectomized rats. Conversion to catechol oestrogens was tested in microsomes from the HPA, pituitary gland and liver, using a catechol-O-methyltransferase-coupled radioenzymatic assay. Oestradiol-17 alpha was the only weak oestrogen receptor ligand. Binding affinities of the other compounds tested were much higher and comparable to those of oestradiol-17 beta. In contrast, oestradiol-17 alpha was rapidly converted to catechol metabolites, while moxestrol was a relatively poor substrate for catechol oestrogen formation. 4-Fluoro-oestradiol could be 2-hydroxylated but not 4-hydroxylated. 2-Fluoro-oestradiol exhibited impaired 2-hydroxylation but normal 4-hydroxylation
— id: 102784, year: 1986, vol: 110, page: 489, stat: Journal Article,

Behavioral and neuroendocrine effects of long-term progesterone treatment in the rat
Biegon A; Parsons B; Krey LC; Kamel F; McEwen BS
1983 Nov;37(5):332-335, Neuroendocrinology
Progesterone and estradiol, alone or in combination, were administered to ovariectomized rats for 2 weeks. Progesterone alone had no effect on body weight, luteinizing hormone (LH) secretion, receptivity or brain cytoplasmic progestin receptors (CPR). Progesterone in combination with estradiol significantly attenuated estrogen suppression of weight gain and estrogen stimulation of receptivity, the LH afternoon surge and induction of CPR, but did not affect the negative feedback of estrogen on morning LH levels. The decrease in CPR after 2 weeks of progesterone is very similar in magnitude to the decrease observed following acute treatment, suggesting that, unlike neurotransmitter agonists and glucocorticoids, progesterone does not cause down regulation of its receptors in the brain following chronic treatment
— id: 58808, year: 1983, vol: 37, page: 332, stat: Journal Article,

Gonadal steroid modulation of LH secretion stimulated by LHRH, Ca2+ and cAMP
Kamel F; Krey LC
1983 Oct;32(2-3):285-300, Molecular & cellular endocrinology
Primary cultures of enzymatically dispersed rat pituitary cells were used to examine steroid effects on LH secretion stimulated by LHRH, Ca2+ and cAMP. Cultures were pretreated for 48 h with testosterone (T) or 17 beta-estradiol (E) and then challenged for 4 h with various secretogogues. T did not affect basal LH secretion; it inhibited the responses to Ca2+ and LHRH; and it potentiated the response to cAMP. E stimulated both basal LH secretion and the responses to all secretogogues, without affecting cell LH content. Thus, T affects stimulus-secretion coupling, while E affects secretion per se. All steroid effects were blocked by steroid antagonists, indicating that steroid action is receptor-mediated regardless of the secretogogue involved. The similarity of steroid effects on the responses to LHRH and Ca2+ but not cAMP suggests that Ca2+ rather than cAMP is a second messenger for LHRH, and that steroid action occurs at some step subsequent to LHRH-stimulated Ca2+ mobilization
— id: 58803, year: 1983, vol: 32, page: 285, stat: Journal Article,

Aromatization: important for testosterone's developmental influences on the neurocircuits mediating cervically stimulated prolactin secretion in the female rat
Krey LC; Kamel F
1983 Sep 5;274(1):188-192, Brain research
Adult female rats treated neonatally with testosterone or estrogen fail to respond to cervical stimulation with episodic prolactin release. The developmental action of testosterone is inhibited by co-administration of 1,4,6-androstatriene-3,17-dione at a dose sufficient to inhibit the testosterone-induced rise in estrogen receptors in limbic brain cell nuclei. These data suggest the importance of aromatization for testosterone action
— id: 58809, year: 1983, vol: 274, page: 188, stat: Journal Article,

Anovulation in female rats induced by neonatal administration of the catechol estrogens, 2-hydroxy-estradiol and 4-hydroxy-estradiol
MacLusky, N J; Riskalla, M; Krey, L; Parvizi, N; Naftolin, F
1983 Nov;37(5):321-327, Neuroendocrinology
The effects of estradiol (E2) and its 2- and 4-hydroxylated metabolites on gonadotrophin regulation in the female rat brain were examined. Neonatal female rats were injected from day 1 through 5 with E2, 2-OHE2 and 4-OHE2, at doses of 0.1, 1 and 10 micrograms/day. At 2, 6 and 24 h after the last estrogen injection, some animals from each treatment group were killed and the concentration of estrogen receptors (ERn) in their brain cell nuclei determined. The remaining animals were allowed to mature. Their vaginal smear patterns were examined from 7 to 9 and from 15 to 17 weeks of age. They were then ovariectomized and tested for their capacity to exhibit a luteinizing hormone (LH) surge in response to estrogen and progesterone injections. In a parallel series of experiments, the affinities of the three test estrogens for alpha-fetoprotein (AFP) were determined from in vitro competition studies with fetal rat serum. All three estrogens increased brain cell nuclear ERn concentrations, measured at 2 h after the final injection. E2 was more potent in this respect than either 4-OHE2 or 2-OHE2. E2 and 4-OHE2 competed for binding to AFP to an approximately equal extent. 2-OHE2, however, was a much weaker competitor for AFP than either of the other two compounds. The neonatal E2 and 4-OHE2 treatments reduced the number of animals showing regular cyclic vaginal smears, at all three doses tested. In contrast, 2-OHE2 significantly affected vaginal cyclicity only at a dose of 10 micrograms/day.(ABSTRACT TRUNCATED AT 250 WORDS)
— id: 102747, year: 1983, vol: 37, page: 321, stat: Journal Article,

Gonadal steroid modulation of LHRH-stimulated LH secretion by pituitary cell cultures
Kamel F; Krey LC
1982 Apr;26(1-2):151-164, Molecular & cellular endocrinology
Enzymatically dispersed rat pituitary cells were grown in primary culture, and LHRH-stimulated LH secretion was measured. Testosterone (T) decreased and 17 beta-estradiol (E) increased pituitary responsiveness to LHRH. The effect of E on LH secretion was partly due to an increase in LH content. There was a latent period of 12 h for E and 18 h for T between the onset of steroid treatment and the manifestation of steroid action. Neither steroid was required to be continuously present in order to exert its effects. After steroid withdrawal, the effect of T persisted for 72 h and that of E for more than 96 h. The actions of both steroids were blocked by protein-synthesis inhibitors. These results are consistent with the hypothesis that steroid effects rely on a mechanism involving alterations in protein synthesis; the affected proteins may be involved in the process of LHRH action
— id: 55830, year: 1982, vol: 26, page: 151, stat: Journal Article,

Intracellular receptors mediate gonadal steroid modulation of LHRH-induced LH release
Kamel F; Krey LC
1982 Nov-Dec;28(3):471-486, Molecular & cellular endocrinology
Primary cultures of enzymatically dispersed rat pituitary cells were used to study the role of intracellular receptors in gonadal steroid modulation of LHRH-induced LH release. Nuclear receptors for both testosterone (T) and 17 beta-estradiol (E) were observed, with KD values of 6.3 and 0.1 nM respectively. Occupation of these receptors was correlated with modulation of LH secretion. The relationship between these two parameters was nonlinear, so that steroid effects on LH secretion were maximal when fewer than 50% of the receptors were occupied. The androgen antagonist cyproterone blocked both T binding to nuclear receptors and T inhibition of LH secretion. Similarly, the estrogen antagonist CI-628 blocked both E binding and E stimulation of LH secretion. In cultures derived from pseudohermaphrodite rats. T did not bind to nuclear receptors, nor did it inhibit LH secretion. These results, showing a relationship between occupation of nuclear receptors and modulation of LH secretion, suggest that steroid effects on LH secretion are mediated by these receptors
— id: 58810, year: 1982, vol: 28, page: 471, stat: Journal Article,

Central and peripheral action of estradiol and catecholestrogens administered at low concentration by constant infusion
Jellinck PH; Krey L; Davis PG; Kamel F; Luine V; Parsons B; Roy EJ; McEwen BS
1981 May;108(5):1848-1854, Endocrinology
— id: 58811, year: 1981, vol: 108, page: 1848, stat: Journal Article,

The catechol estrogens
MacLusky, N J; Naftolin, F; Krey, L C; Franks, S
1981 Dec;15:111-124, Journal of steroid biochemistry
— id: 102732, year: 1981, vol: 15, page: 111, stat: Journal Article,

Parameters of neuroendocrine aromatization and estrogen receptor occupation in the male rat
Krey LC; Kamel F; McEwen BS
1980 Jul 7;193(1):277-283, Brain research
— id: 58812, year: 1980, vol: 193, page: 277, stat: Journal Article,