Gonadotrophin releasing hormone agonist (GnRHa) trigger is effective in the induction of oocyte m... more Gonadotrophin releasing hormone agonist (GnRHa) trigger is effective in the induction of oocyte maturation and prevention of ovarian hyperstimulation syndrome during IVF treatment. This trigger concept, however, results in early corpora lutea demise and consequently luteal phase dysfunction and impaired endometrial receptivity. The aim of this strenghths, weaknesses, opportunities and threats analysis was to summarize the progress made over the past 15 years to optimize ongoing pregnancy rates after GnRHa trigger. The advantages and potential drawbacks of this type of triggering are reviewed. The current approach to the management of GnRHa trigger in autologous cycles is based on the peak serum oestradiol level or follicle number and aims at a fresh embryo transfer or a segmentation approach with elective cryopreservation policy. We recommend intensive luteal support with transdermal oestradiol and intramuscular progesterone alone if peak serum oestradiol is 4000 or more pg/ml after GnRHa trigger or dual trigger with GnRHa and HCG 1000 IU if peak serum oestradiol is less than 4000 pg/mL. On the contrary, we recommend HCG 1500 IU 35 h after GnRHa trigger if there are less than 25 follicles, or freeze all oocytes or embryos if there are over 25 follicles.
To assess the occult pregnancy rate after "negative" first post-embryo transfer (ET) se... more To assess the occult pregnancy rate after "negative" first post-embryo transfer (ET) serum β-hCG results. Two-part retrospective cohort study and nested case series. University-based fertility center. A total of 1,571 negative first post-ET serum β-hCG results were included in the study; 1,326 results (primary cohort, June 2009-December 2013) were initially reported as <5 mIU/mL and 245 results (secondary cohort, January 2014-March 2015) were reported as discrete values from 1.0 to 5.0 mIU/mL. None. Rates of occult pregnancy, ectopic pregnancy, and complications after negative first post-ET serum β-hCG results. A total of 88.8% (1,178/1,326) of the negative first post-ET results reported as <5 were actually <1.0 mIU/mL. Occult pregnancy was incidentally identified in 1.2% (12/1,041) of subjects with follow-up. Six had ectopic pregnancies, and seven experienced serious complications; 11 (91.7%) of the 12 occult pregnancies had a first post-ET serum β-hCG level of 1...
The effects of cortisol on steroid secretion by human luteinized granulosa cells was investigated... more The effects of cortisol on steroid secretion by human luteinized granulosa cells was investigated by culturing cells from mature follicles of gonadotropin-treated women. After culturing for 24 hours in medium containing 10% fetal bovine serum, the cells were washed and incubated in a chemically defined medium. Cortisol (10(-5), 10(-6), and 10(-7) M), follicle-stimulating hormone (FSH; 100 ng/ml), FSH plus cortisol (10(-6], or vehicle were added to replicate cultures. After incubating for 6 hours, medium was harvested and assayed for estradiol (E2) and progesterone (P). Cortisol (10(-5), 10(-6) but not 10(-7) M) significantly (P less than 0.05) stimulated E2 secretion when compared to controls. FSH alone did not alter E2 secretion while FSH plus cortisol had a similar effect to cortisol alone. Likewise, P secretion was significantly (P less than 0.05) increased by cortisol when compared with controls and the stimulation was dose dependent. FSH also stimulated P secretion, but the combination of FSH and cortisol did not give an additive or synergistic effect. It appears that cortisol can stimulate the secretion of E2 and P by human granulosa cells without involving FSH receptor interaction. The findings suggest that, in addition to pituitary and ovarian hormones, cortisol may also be directly involved in granulosa cell function.
To determine aneuploidy for chromosome 16 by recycling nuclei of cells already analyzed for chrom... more To determine aneuploidy for chromosome 16 by recycling nuclei of cells already analyzed for chromosomes X, Y, 18, 13, and 21 using multiple fluorescence in situ hybridization in preimplantation human embryos in a time frame compatible with clinical IVF and to asssess the incidence of chromosome 16 aneuploidy in embryos related to maternal age. Prospective experimental study. In vitro fertilization program in a tertiary center. One hundred four consenting patients undergoing IVF. Chromosome 16 ploidy was analyzed in a total of 195 embryos. In 89 embryos, a standard multiple-probe fluorescence in situ hybridization was used for chromosomes X, Y, 18 and 16 (series 1). The remaining 106 embryos (series 2) were reanalyzed with a new procedure for chromosome 16, which involves rehybridization with a digoxigenin-labeled alpha satellite probe after the standard analysis for chromosomes X, Y, 18, 13, and 21 was completed. The embryos were assigned to one of three groups according to the women&#39;s age; group 1: &lt;/= 34 years (n = 34), group 2: 35 to 39 years (n = 47); group 3: &gt;/= 40 years (n = 23). Successful analysis, including biopsy, fixation, and fluorescence in situ hybridization was achieved in 86% of the blastomeres within approximately 10 hours. A significant relationship was found between the rate of aneuploidy for chromosome 16 and increasing maternal age: group 1: 0%, group 2: 6.3%, and group 3: 11.7%. Monosomy for chromosome 16 was found in 72.7% of the 11 embryos carrying chromosome 16 anomalies, with the remaining three embryos having two trisomies and one tetrasomy. This new protocol was applied clinically to five patients undergoing preimplantation aneuploidy assessment. Aneuploidy for chromosome 16 was found in five embryos from three of those patients. This study demonstrates that preimplantation genetic diagnosis of the major human aneuploidies is achievable within a time frame compatible with IVF. In addition, this study confirms, for embryos, the existing data from spontaneous abortions suggesting that chromosome 16 aneuploidy increases with maternal age. The high prevalence of embryonic monosomy, which is rarely found in spontaneous abortions, suggests that monosomy 16 could be a factor associated with failure of implantation, as well as pointing to a different mechanism involved in the generation of chromosome 16 aneuploidy.
The main conclusion of this study is that a profound suppression of the pituitary and ovary can b... more The main conclusion of this study is that a profound suppression of the pituitary and ovary can be associated with an inadequate response which may require a longer or different regimen of stimulation to achieve the desired outcome for IVF. We suggest that a pretreatment determination of E2 and gonadotropins can be of value to predict the nature of ovarian response in women with suppressed pituitary-ovarian function.
The purpose of the present study was to determine whether the presence of one or more multinuclea... more The purpose of the present study was to determine whether the presence of one or more multinucleated blastomeres during early embryonic development is associated with chromosomal abnormalities in sibling blastomeres of that embryo. Embryos with multinucleated cells (n = 47) detected on day 2 or 3 or development were compared to dividing embryos without multinucleation. Arrested embryos were excluded from this study. Chromosome abnormalities were detected using fluorescent in-situ hybridization (FISH) with X, Y, 18 and 13/21 chromosome-specific probes. Of 47 embryos included in this study, 76.6% were chromosomally abnormal, compared to 50.9% in the control group (P < 0.001). Excluding aneuploidy, which is originated in the gametes and not the embryo, the differences were even higher, with 74.5% of multinucleated embryos being chromosomally abnormal compared to 32.3% of non-multinucleated embryos (P < 0.001). Day of multinucleation appearance, number of nuclei per cell, number o...
The purpose of the present study was to determine whether the presence of one or more multinuclea... more The purpose of the present study was to determine whether the presence of one or more multinucleated blastomeres during early embryonic development is associated with chromosomal abnormalities in sibling blastomeres of that embryo. Embryos with multinucleated cells (n = 47) detected on day 2 or 3 or development were compared to dividing embryos without multinucleation. Arrested embryos were excluded from this study. Chromosome abnormalities were detected using fluorescent in-situ hybridization (FISH) with X, Y, 18 and 13/21 chromosome-specific probes. Of 47 embryos included in this study, 76.6% were chromosomally abnormal, compared to 50.9% in the control group (P < 0.001). Excluding aneuploidy, which is originated in the gametes and not the embryo, the differences were even higher, with 74.5% of multinucleated embryos being chromosomally abnormal compared to 32.3% of non-multinucleated embryos (P < 0.001). Day of multinucleation appearance, number of nuclei per cell, number o...
American Journal of Obstetrics and Gynecology, 1988
Steroid secretion and structure of granulosa cells on floating collagen gels were compared with t... more Steroid secretion and structure of granulosa cells on floating collagen gels were compared with those of cells grown on plastic. Granulosa cells from follicles of gonadotropin-treated women were plated either onto dishes coated with type I collagen or onto plastic dishes. Medium containing serum was removed after 24 hours, defined medium was added, and the gel was floated. Medium was changed daily for 3 days, after which the granulosa cells were prepared for light and electron microscopy. Cells grown on collagen secreted significantly more estradiol and progesterone than those grown on plastic during the 3 days of culture. The round multilayered granulosa cells on collagen had abundant mitochondria and lipid droplets and they formed numerous intercellular junctions. On plastic surfaces, flat granulosa cells grew as a monolayer with few junctions and less abundant mitochondria or lipid droplets. We conclude that growth on floating collagen promotes structural changes of human granulosa cells that enhances cell interaction and secretion of steroid hormones.
Among patients with advanced stage serous borderline tumors of the ovary, those with micropapilla... more Among patients with advanced stage serous borderline tumors of the ovary, those with micropapillary architecture or invasive implants have the greatest risk of malignant transformation. In the absence of these patterns, consideration can be given to preservation of reproductive function. A 28-year-old, nulliparous patient presented with symptoms mimicking advanced ovarian cancer. Histology showed a serous borderline tumor with a hierarchical branching pattern. Surgery was able to remove all visible disease but still preserve the uterus and a portion of one ovary. She subsequently underwent in vitro fertilization and delivered a full-term infant.
Luteal phase synchronization of follicular growth has been suggested as a means to improve ovaria... more Luteal phase synchronization of follicular growth has been suggested as a means to improve ovarian response in low responders. We compared luteal E2 and antagonist (n=256) with luteal E2 only (n=57) before antagonist protocol in low responders. The addition of GnRH antagonist to luteal E2 for luteal suppression before ovarian stimulation for IVF does not improve IVF outcomes in poor responders.
Gonadotrophin releasing hormone agonist (GnRHa) trigger is effective in the induction of oocyte m... more Gonadotrophin releasing hormone agonist (GnRHa) trigger is effective in the induction of oocyte maturation and prevention of ovarian hyperstimulation syndrome during IVF treatment. This trigger concept, however, results in early corpora lutea demise and consequently luteal phase dysfunction and impaired endometrial receptivity. The aim of this strenghths, weaknesses, opportunities and threats analysis was to summarize the progress made over the past 15 years to optimize ongoing pregnancy rates after GnRHa trigger. The advantages and potential drawbacks of this type of triggering are reviewed. The current approach to the management of GnRHa trigger in autologous cycles is based on the peak serum oestradiol level or follicle number and aims at a fresh embryo transfer or a segmentation approach with elective cryopreservation policy. We recommend intensive luteal support with transdermal oestradiol and intramuscular progesterone alone if peak serum oestradiol is 4000 or more pg/ml after GnRHa trigger or dual trigger with GnRHa and HCG 1000 IU if peak serum oestradiol is less than 4000 pg/mL. On the contrary, we recommend HCG 1500 IU 35 h after GnRHa trigger if there are less than 25 follicles, or freeze all oocytes or embryos if there are over 25 follicles.
To assess the occult pregnancy rate after "negative" first post-embryo transfer (ET) se... more To assess the occult pregnancy rate after "negative" first post-embryo transfer (ET) serum β-hCG results. Two-part retrospective cohort study and nested case series. University-based fertility center. A total of 1,571 negative first post-ET serum β-hCG results were included in the study; 1,326 results (primary cohort, June 2009-December 2013) were initially reported as <5 mIU/mL and 245 results (secondary cohort, January 2014-March 2015) were reported as discrete values from 1.0 to 5.0 mIU/mL. None. Rates of occult pregnancy, ectopic pregnancy, and complications after negative first post-ET serum β-hCG results. A total of 88.8% (1,178/1,326) of the negative first post-ET results reported as <5 were actually <1.0 mIU/mL. Occult pregnancy was incidentally identified in 1.2% (12/1,041) of subjects with follow-up. Six had ectopic pregnancies, and seven experienced serious complications; 11 (91.7%) of the 12 occult pregnancies had a first post-ET serum β-hCG level of 1...
The effects of cortisol on steroid secretion by human luteinized granulosa cells was investigated... more The effects of cortisol on steroid secretion by human luteinized granulosa cells was investigated by culturing cells from mature follicles of gonadotropin-treated women. After culturing for 24 hours in medium containing 10% fetal bovine serum, the cells were washed and incubated in a chemically defined medium. Cortisol (10(-5), 10(-6), and 10(-7) M), follicle-stimulating hormone (FSH; 100 ng/ml), FSH plus cortisol (10(-6], or vehicle were added to replicate cultures. After incubating for 6 hours, medium was harvested and assayed for estradiol (E2) and progesterone (P). Cortisol (10(-5), 10(-6) but not 10(-7) M) significantly (P less than 0.05) stimulated E2 secretion when compared to controls. FSH alone did not alter E2 secretion while FSH plus cortisol had a similar effect to cortisol alone. Likewise, P secretion was significantly (P less than 0.05) increased by cortisol when compared with controls and the stimulation was dose dependent. FSH also stimulated P secretion, but the combination of FSH and cortisol did not give an additive or synergistic effect. It appears that cortisol can stimulate the secretion of E2 and P by human granulosa cells without involving FSH receptor interaction. The findings suggest that, in addition to pituitary and ovarian hormones, cortisol may also be directly involved in granulosa cell function.
To determine aneuploidy for chromosome 16 by recycling nuclei of cells already analyzed for chrom... more To determine aneuploidy for chromosome 16 by recycling nuclei of cells already analyzed for chromosomes X, Y, 18, 13, and 21 using multiple fluorescence in situ hybridization in preimplantation human embryos in a time frame compatible with clinical IVF and to asssess the incidence of chromosome 16 aneuploidy in embryos related to maternal age. Prospective experimental study. In vitro fertilization program in a tertiary center. One hundred four consenting patients undergoing IVF. Chromosome 16 ploidy was analyzed in a total of 195 embryos. In 89 embryos, a standard multiple-probe fluorescence in situ hybridization was used for chromosomes X, Y, 18 and 16 (series 1). The remaining 106 embryos (series 2) were reanalyzed with a new procedure for chromosome 16, which involves rehybridization with a digoxigenin-labeled alpha satellite probe after the standard analysis for chromosomes X, Y, 18, 13, and 21 was completed. The embryos were assigned to one of three groups according to the women&#39;s age; group 1: &lt;/= 34 years (n = 34), group 2: 35 to 39 years (n = 47); group 3: &gt;/= 40 years (n = 23). Successful analysis, including biopsy, fixation, and fluorescence in situ hybridization was achieved in 86% of the blastomeres within approximately 10 hours. A significant relationship was found between the rate of aneuploidy for chromosome 16 and increasing maternal age: group 1: 0%, group 2: 6.3%, and group 3: 11.7%. Monosomy for chromosome 16 was found in 72.7% of the 11 embryos carrying chromosome 16 anomalies, with the remaining three embryos having two trisomies and one tetrasomy. This new protocol was applied clinically to five patients undergoing preimplantation aneuploidy assessment. Aneuploidy for chromosome 16 was found in five embryos from three of those patients. This study demonstrates that preimplantation genetic diagnosis of the major human aneuploidies is achievable within a time frame compatible with IVF. In addition, this study confirms, for embryos, the existing data from spontaneous abortions suggesting that chromosome 16 aneuploidy increases with maternal age. The high prevalence of embryonic monosomy, which is rarely found in spontaneous abortions, suggests that monosomy 16 could be a factor associated with failure of implantation, as well as pointing to a different mechanism involved in the generation of chromosome 16 aneuploidy.
The main conclusion of this study is that a profound suppression of the pituitary and ovary can b... more The main conclusion of this study is that a profound suppression of the pituitary and ovary can be associated with an inadequate response which may require a longer or different regimen of stimulation to achieve the desired outcome for IVF. We suggest that a pretreatment determination of E2 and gonadotropins can be of value to predict the nature of ovarian response in women with suppressed pituitary-ovarian function.
The purpose of the present study was to determine whether the presence of one or more multinuclea... more The purpose of the present study was to determine whether the presence of one or more multinucleated blastomeres during early embryonic development is associated with chromosomal abnormalities in sibling blastomeres of that embryo. Embryos with multinucleated cells (n = 47) detected on day 2 or 3 or development were compared to dividing embryos without multinucleation. Arrested embryos were excluded from this study. Chromosome abnormalities were detected using fluorescent in-situ hybridization (FISH) with X, Y, 18 and 13/21 chromosome-specific probes. Of 47 embryos included in this study, 76.6% were chromosomally abnormal, compared to 50.9% in the control group (P < 0.001). Excluding aneuploidy, which is originated in the gametes and not the embryo, the differences were even higher, with 74.5% of multinucleated embryos being chromosomally abnormal compared to 32.3% of non-multinucleated embryos (P < 0.001). Day of multinucleation appearance, number of nuclei per cell, number o...
The purpose of the present study was to determine whether the presence of one or more multinuclea... more The purpose of the present study was to determine whether the presence of one or more multinucleated blastomeres during early embryonic development is associated with chromosomal abnormalities in sibling blastomeres of that embryo. Embryos with multinucleated cells (n = 47) detected on day 2 or 3 or development were compared to dividing embryos without multinucleation. Arrested embryos were excluded from this study. Chromosome abnormalities were detected using fluorescent in-situ hybridization (FISH) with X, Y, 18 and 13/21 chromosome-specific probes. Of 47 embryos included in this study, 76.6% were chromosomally abnormal, compared to 50.9% in the control group (P < 0.001). Excluding aneuploidy, which is originated in the gametes and not the embryo, the differences were even higher, with 74.5% of multinucleated embryos being chromosomally abnormal compared to 32.3% of non-multinucleated embryos (P < 0.001). Day of multinucleation appearance, number of nuclei per cell, number o...
American Journal of Obstetrics and Gynecology, 1988
Steroid secretion and structure of granulosa cells on floating collagen gels were compared with t... more Steroid secretion and structure of granulosa cells on floating collagen gels were compared with those of cells grown on plastic. Granulosa cells from follicles of gonadotropin-treated women were plated either onto dishes coated with type I collagen or onto plastic dishes. Medium containing serum was removed after 24 hours, defined medium was added, and the gel was floated. Medium was changed daily for 3 days, after which the granulosa cells were prepared for light and electron microscopy. Cells grown on collagen secreted significantly more estradiol and progesterone than those grown on plastic during the 3 days of culture. The round multilayered granulosa cells on collagen had abundant mitochondria and lipid droplets and they formed numerous intercellular junctions. On plastic surfaces, flat granulosa cells grew as a monolayer with few junctions and less abundant mitochondria or lipid droplets. We conclude that growth on floating collagen promotes structural changes of human granulosa cells that enhances cell interaction and secretion of steroid hormones.
Among patients with advanced stage serous borderline tumors of the ovary, those with micropapilla... more Among patients with advanced stage serous borderline tumors of the ovary, those with micropapillary architecture or invasive implants have the greatest risk of malignant transformation. In the absence of these patterns, consideration can be given to preservation of reproductive function. A 28-year-old, nulliparous patient presented with symptoms mimicking advanced ovarian cancer. Histology showed a serous borderline tumor with a hierarchical branching pattern. Surgery was able to remove all visible disease but still preserve the uterus and a portion of one ovary. She subsequently underwent in vitro fertilization and delivered a full-term infant.
Luteal phase synchronization of follicular growth has been suggested as a means to improve ovaria... more Luteal phase synchronization of follicular growth has been suggested as a means to improve ovarian response in low responders. We compared luteal E2 and antagonist (n=256) with luteal E2 only (n=57) before antagonist protocol in low responders. The addition of GnRH antagonist to luteal E2 for luteal suppression before ovarian stimulation for IVF does not improve IVF outcomes in poor responders.
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Papers by Claudio Benadiva