Mitochondrial deletions in luteinized granulosa cells as a function of age in women undergoing in vitro fertilization

OBJECTIVE: To test the hypothesis that mitochondrial DNA (mtDNA) deletions are more prevalent in granulosa cells from women of advanced reproductive age than from younger women undergoing IVF. DESIGN: Granulosa cells screened for presence or absence of the 4977-bp deletion in human mtDNA. SETTING: University-based fertility clinic. PATIENT(S): Twenty-four women divided equally between two groups: /=38 years old. INTERVENTION(S): Patients were given gonadotropin stimulation in preparation for IVF with granulosa cells isolated at the time of follicular aspiration. MAIN OUTCOME MEASURE(S): Presence or absence of the 4977-bp deletion in human mtDNA. RESULT(S): Seven out of 12 women analyzed who were <38 years old and 0 out of 12 women who were >38 years old had normal mtDNA as indicated by the presence of the 4977-bp fragment. CONCLUSION(S): These data suggest that women over the age of 38 have granulosa cells that contain a substantial decrease in the level of normal mitochondria as compared with women 相似文献   

Simultaneous laparoscopy for endometriotic women undergoing in vitro fertilization

ObjectiveThe aim of this study was to investigate simultaneous laparoscopy in endometriotic women with infertility undergoing in vitro fertilization (IVF).Materials and MethodsForty-seven infertile patients with endometriosis were enrolled in this retrospective study and underwent IVF cycles in a university affiliated hospital.ResultsThe chemical pregnancy, clinical pregnancy and live birth rates were statistically significantly different between patients with minimal or mild stage endometriosis and patients with moderate or severe stage endometriosis, who received simultaneous laparoscopy and modified IVF with a GnRH antagonist protocol. A higher live birth rate was achieved in IVF patients with minimal or mild stage endometriosis combined with laparoscopic treatment, than in patients who received traditional IVF with prior laparoscopic surgery for endometrioma.ConclusionSimultaneous laparoscopy combined with a modified IVF (GnRH antagonist) protocol may benefit patients with minimal and mild endometriosis. Traditional GnRH agonist IVF cycles may improve the fecundity rates in women with moderate and severe endometriosis after laparoscopic treatment.     

Mitochondrial gene expression in granulosa cells of severe endometriosis with in vitro fertilization and embryo transfer

The expression of mitochondrial genes in granulosa cells was quantitated by real-time polymerase chain reaction. The expression ratio of mitochondrial genes in granulosa cells of patients with severe endometriosis showed no statistically significant difference compared with cases of tubal infertility without endometriosis.     

DNA ploidy of human granulosa cells from natural and stimulated in vitro fertilization cycles

OBJECTIVE: To analyze and compare the DNA ploidy of granulosa cells from natural and gonadotropin-stimulated follicles obtained during IVF. DESIGN: Retrospective analysis of laboratory data. SETTING: University medical center. PATIENT(S): Seventy-three aspirates of dominant follicles from natural IVF cycles and 113 aspirates from gonadotropin-stimulated cycles were analyzed. INTERVENTION(S): Cytospins were prepared and stained by the Feulgen-thionine method. MAIN OUTCOME MEASURE(S): Image DNA analysis was performed on an automated high-resolution image cytometer. DNA content and the number of nuclei with DNA content >5c were measured. RESULT(S): All samples from natural and gonadotropin-stimulated follicles were found to be diploid. Single cells with DNA content >5c were found in follicular fluid samples of four women with natural IVF cycles and in samples of nine women with gonadotropin-stimulated cycles. CONCLUSION(S): DNA ploidy of granulosa cells from natural follicles has not been studied before. In natural samples, granulosa cells were only diploid, without euploid polyploidization. We were unable to confirm DNA aneuploidy of granulosa cells in gonadotropin-stimulated follicles of women undergoing IVF.     

Fluoxetine treatment for anxiety in women undergoing in vitro fertilization

Objective

To compare anxiety levels experienced during 4 stressful periods of in vitro fertilization (IVF) and treatment outcomes between women taking fluoxetine and a placebo.

Methods

A prospective, randomized, double-blind, placebo-controlled trial of patients allocated to receive either fluoxetine (FLX) or folic acid (FA). Anxiety state was assessed at the beginning of ovarian stimulation (OS), ovum pick-up, embryo transfer, and on the day of the pregnancy test (DPT) using the State-Trait Anxiety Inventory (STAI).

Results

Baseline STAI-S and STAI-T were normal. From OS to DPT, STAI-S increased from 42.8 ± 10.6 to 44 ± 9.0 in the FLX group and from 40.9 ± 8.1 to 45.3 ± 8.3 in the FA group (P = 0.03 and P = 0.001, respectively). IVF outcome was not affected by the treatment in the two groups.

Conclusions

Caution is needed in prescribing fluoxetine to alleviate anxiety in patients undergoing IVF. Studies are needed to determine whether other selective serotonin reuptake inhibitors or higher fluoxetine doses can relieve emotional distress without affecting IVF outcome.     

Endometrial integrin expression in women undergoing in vitro fertilization and the association with subsequent treatment outcome

OBJECTIVE: To study the endometrial expression of three integrins (alpha v beta 3, alpha 4 beta 1, and alpha 1 beta 1) in women undergoing IVF-intracytoplasmic sperm injection (ICSI) treatment and assess whether they could be used to predict subsequent treatment success.Prospective observational study.Healthy volunteers in a large teaching hospital. PATIENT(S): Sixty-six patients attending for IVF-ICSI treatment. INTERVENTION(S): Timed endometrial biopsies were taken, during the implantation window at LH + 7-9 days, from women before IVF-ICSI treatment. MAIN OUTCOME MEASURE(S): Histological dating of endometrium and immunohistochemical staining intensity of alpha 4 beta 1, alpha v beta 3, and alpha 1 beta 1 integrins. The integrin levels were correlated with subsequent success rates. RESULT(S): There was a statistically significantly greater expression of alpha v beta 3 in the luminal epithelium of those patients who had successful treatment. However, treatment was successful in some patients with negative expression. CONCLUSION(S): Integrins are important markers of endometrial receptivity. There is an association between an in-phase endometrial biopsy, with positive luminal alpha v beta 3 integrin expression, and subsequent treatment success. However, the clinical value of assessing the endometrium before treatment has drawbacks, and further work needs to be done before this can be considered a clinically useful test.     

Anomalies of human oocytes from infertile women undergoing treatment by in vitro fertilization

During in vitro fertilization treatment, three types of oocyte anomalies were observed in four infertile women. These anomalies were related to (1) failure of oocyte maturation from the germinal vesicle stage of meiosis, (2) failure of polar body formation and cleavage, and (3) absence of oocytes in otherwise mature follicular aspirates. These anomalies are considered to contribute to these patients' infertility.     

Oocyte morphological abnormalities in overweight women undergoing in vitro fertilization cycles

The effect of elevated body mass index (BMI) on the oocyte quality was investigated in women undergoing in vitro fertilization (IVF) cycles. A total of 268 patients classified on the basis of BMI subject to the first reproductive treatment were included in this study: the normal weight (NW) group consisted of 160 patients with BMI 19–24.9?kg/m² and the overweight (OW) group consisted of 108 patients with BMI ≥25?kg/m². All women were treated with a standard long luteal protocol. The oocyte features were classified as extracytoplasmic or cytoplasmic abnormalities. Outcomes were oocyte morphology, embryo quality, fertilization and implantation rates, and the ovarian response to stimulation. A higher percentage of oocytes with granular cytoplasm was found in women with BMI ≥25 (p?=?0.04). However, percentages of mature, immature oocytes and germinal vesicle were similar in both groups. No differences were found in fertilization and cleavage rates and percentages of embryo quality. The implantation rate (p?<?0.001) was significantly lower in the OW group than in the NW group. The amount of gonadotrophins was significantly higher in OW group (p?=?0.003). These findings suggest that the poor reproductive outcome of obese women is influenced by the release of ova with reduced fertilization potential.     

Cell-free DNA and telomere length among women undergoing in vitro fertilization treatment

Purpose

The current research is aimed at finding potential non-invasive bio-markers that will help us learn more about the mechanisms at play in failed assisted reproduction treatment. This exploratory pilot study examined the relationship between cell-free DNA (CFD) in plasma and telomere length in lymphocytes among women undergoing in vitro fertilization (IVF) and compared telomere length and CFD levels to a healthy control group.

Methods

Blood of 20 women undergoing IVF was collected at three time points during the IVF cycle. We assessed the relationship between CFD and telomere length as well as controlling for morning cortisol levels. We also collected blood of 10 healthy controls at two time points (luteal and follicular phases of the menstrual cycle) and compared mean telomere length, CFD, and cortisol levels between the IVF patients and healthy controls.

Results

The results revealed an inverse relationship between CFD levels and telomere lengths at several time points that remained significant even after controlling for cortisol levels. Women undergoing IVF had statistically significant higher levels of CFD and shorter telomeres compared to healthy controls.

Conclusions

The relationship between telomere length and CFD should be further explored in larger studies in order to uncover potential mechanisms that cause both shortened telomere length and elevated CFD in women undergoing IVF.     

Glycosaminoglycans in follicular fluid from women undergoing in vitro fertilization and their relationship to cumulus expansion, fertilization, and development

The glycosaminoglycans (GAGs) chondroitin sulfate (CS) and heparan sulfate (HS) were assayed in fluid from 178 individual follicles obtained from women after human menopausal gonadotropin ovulation induction for subsequent in vitro fertilization. CS and HS concentrations ranged from 0 to 3.9 and 0 to 18.6 mg/ml, respectively. The percentage of oocytes fertilized was significantly higher when CS concentrations ranged from 0.3 to 0.8 mg/ml (P less than 0.03). As HS levels increased, maturation scores by visual assessment of the egg-corona-cumulus cell complexes increased (P less than 0.05), but the percentage of oocytes fertilized was not affected. Cleavage rates of developing embryos were not related to the concentrations of GAGs.     

Useful predictors of ovarian stimulation response in women undergoing in vitro fertilization

Eighty-seven patients undergoing in vitro fertilization were studied to evaluate the prognostic value of different tests in predicting ovarian stimulation response. We studied basal follicle-stimulating hormone (FSH) value on cycle day 3, the clomiphene citrate (CC) challenge test and serum estradiol levels on cycle day 3. Patients with elevated basal FSH levels needed more ampoules of gonadotropins for stimulation and had a higher cancellation rate because of poor response. Patients with abnormal CC challenge test results also needed more ampoules of gonadotropins, but the higher cancellation rate in this group did not reach statistical significance. The basal estradiol level did not contribute to further discrimination between groups. We conclude that basal FSH level on cycle day 3 is a useful prognosticator of ovarian stimulation response.     

Difference in mitochondrial gene expression in granulosa cells between recombinant FSH and hMG cycles under in vitro fertilization and transfer

Purpose

Examination of the mitochondrial mRNA expression in granulosa cells from an unspecified population of infertile patients to evaluate whether recombinant follicle stimulating hormone (recFSH) is more effective in producing higher quality embryo rates compared with human menopausal gonadotropin (hMG).

Method

Thirty-nine patients who underwent the in vitro fertilization and embryo transfer program were retrospectively examined. Patients were administered recFSH (n = 18) or hMG (n = 20) in a long protocol where GnRH agonist was used. Granulosa cells were obtained during oocyte retrieval and examined for mitochondria mRNA expression ratio against GAPDH. Expressions of mitochondria mRNA were evaluated by real-time PCR analysis.

Results

The high-quality embryo rate in the hMG cycle was higher than in the recFSH cycle, and the total dose of hMG showed a positive correlation with the expression level of mitochondrial genes in granulosa cells. Moreover, mitochondria mRNA expression was higher in the hMG cycle than in the recFSH cycle.

Conclusions

Compared with recFSH, hMG induces a higher mitochondrial gene expression ratio in granulosa cells at the time of oocyte retrieval and, therefore, may lead to higher quality embryo rates.     

A prospective study of stress among women undergoing in vitro fertilization or gamete intrafallopian transfer

OBJECTIVE: To evaluate whether baseline or procedural stress during in vitro fertilization (IVF) or gamete intrafallopian transfer (GIFT) affects pregnancy or live birth delivery rates. DESIGN: Prospective study. SETTING: Seven clinics in Southern California between 1993 and 1998.Patient(s): One hundred and fifty-one women completed two questionnaires. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The number of oocytes aspirated and fertilized, the number of embryos transferred, the achievement of a pregnancy, live birth delivery, and infant outcomes. RESULT(S): Positive-affect negative-affect score at baseline negatively influenced the number of oocytes retrieved and embryos transferred. A higher expectation of pregnancy was associated with greater numbers of oocytes fertilized and embryos transferred. At baseline, the risk of no live birth was 93% lower for women who had the highest positive-affect score compared to those with the lowest score. Furthermore, the score on the Infertility Reaction Scale was related to negative outcomes in live birth delivery, infant birth weight, and multiple births. During the time of the procedure, the PANAS and Bipolar Profile of Moods States results were related to the number of oocytes fertilized and embryos transferred; stress did not affect pregnancy or delivery. CONCLUSION(S): Baseline (acute and chronic) stress affected biologic end points (i.e., number of oocytes retrieved and fertilized), as well as pregnancy, live birth delivery, birth weight, and multiple gestations, whereas (procedural) stress only influenced biologic end points.     

Leptin affects pregnancy outcome of in vitro fertilization and steroidogenesis of human granulosa cells

Purpose: This study was to examine the serum leptin levels in the prediction of pregnancy outcomes in women receiving ovarian hyperstimulation. Effect of leptin on the steroidogenesis was evaluated for granulosa cell (GC) culture in vitro. Method: Serum levels of leptin and estradiol were measured on Day 2, the time of hCG administration and oocyte retrieval in 50 women undergoing long-course GnRH agonist ovarian hyperstimulation. The production of estrogen and progesterone in granulosa cell culture were detected after administration of leptin at the absence or presence of FSH 1 mIU. Results: Leptin levels at the time of hCG injection were significantly lower in the pregnant women than in those without pregnancy. Estradiol concentrations were not correlated with leptin levels during the time of hCG injection and oocyte retrieval. High leptin concentration suppressed both basal estradiol and progesterone productions in GC. Leptin in high doses suppressed estradiol production, but did not alter progesterone production of GC in the presence of FSH. Conclusions: Serum leptin levels may be predictive of IVF pregnancy outcome with the effect on intraovarian progesterone/estradiol ratio during the follicular phase. Significantly low serum leptin levels were noted in the pregnant women than in the nonpregnant women.     

11 beta-Hydroxysteroid dehydrogenase enzyme activity in granulosa cells derived from ovarian follicles of in vitro fertilization patients

11 beta-Hydroxysteroid dehydrogenase (HSD) activity was measured in freshly frozen granulosa cells isolated from follicles of twenty-one infertility patients undergoing in vitro fertilization-embryo transfer (IVF-ET). A total of 213 follicles were analyzed for 11 beta-HSD activity. Both nicotinamide-adenine dinucleotide (NAD) and nicotinamide-adenine dinucleotide phosphate (NADP) dependent 11 beta-dehydrogenase activities were measured in granulosa cells. The activity in reductive direction (11-oxoreductase activity) was not measurable either with NADH or NADPH as cofactors. NAD- and NADP-dependent dehydrogenase activities are in comparable levels at 100 nmol/l and 1 mumol/l corticosteroid concentrations. For comparing enzyme activities of individual follicles, significant enzyme activity was considered to be a level of > 2 nmol/l/min/mg. 41.3% of the follicles demonstrated enzyme activity, 58.7% did not. The mean 11 beta-dehydrogenase (11 beta-DH) activity was calculated for each patient. Those patients with levels > 2 nmol/l/min/mg were considered enzyme positive; those with mean levels < 2 nmol/l/min/mg were considered negative. No significant association was noted between follicle size, oocyte maturity or fertilization rates and 11 beta-DH activity. This study noted the presence of 11 beta-DH activity in granulosa cells, however, no association with oocyte maturity and fertilization was found.