Hormone levels of follicular fluids with and without oocytes in patients who received gonadotropin-releasing hormone analogues and gonadotropins in an in vitro fertilization program

Background Are follicles where no oocytes are retrieved empty follicles?Methods The levels of estradiol (E₂), progesterone (P), testosterone (T), cortisol (F), and prolactin (PRL) of follicular fluids (FF) aspirated individually from 34 randomly selected IVF patients in whom no oocytes were recovered were compared with the respective hormone levels of FF obtained from the same patients when oocytes were retrieved. Two FF without oocytes of a 35th patient in whom no oocytes were retrieved were analyzed.Results Hormones did not differ significantly in the paired samples, while in the two FF of the 35th woman they were in agreement with cystic follicles.Conclusions It is necessary to differentiate aspirated follicles where no oocytes are retrieved from the empty follicle syndrome, which was not observed in the IVF series studied and should be rare in IVF patients.     

Human follicular fluid proteoglycans in relation to in vitro fertilization

Objective: To determine whether the concentrations of proteoglycans and hyaluronan in human follicular fluid (FF) are associated with follicular volume, oocyte fertilization, and ET during IVF.Design: The FF from individual follicles were collected. Enzyme-linked immunosorbent assay methods for quantification of a larger chondroitin sulfate proteoglycan and a smaller composite heparan-chondroitin sulfate proteoglycan were established. Hyaluronan and E₂ were measured by RIA techniques.Patient(s): Sixteen infertile women participating in the IVF program.Main Outcome Measure(s): Concentrations of the proteoglycans, follicular volume, fertilization, and ET rates.Result(s): The follicles contained high concentrations of proteoglycans with an average of 0.8 mg/mL of FF, and approximately 70% consisted of the larger chondroitin sulfate proteoglycan, and 30% of the heparan-chondroitin sulfate proteoglycan. A negative correlation was found between the follicular volume, the chondroitin sulfate proteoglycan (r = −0.43), and hyaluronan (r = −0.56). The percentage of embryos developed in culture was significantly higher in follicles larger than 2 mL. A significant and 35% lower concentration of the chondroitin sulfate proteoglycan was found in larger follicles from which subsequent ET was observed. The heparan-chondroitin sulfate proteoglycan and hyaluronan were both unrelated to fertilization and ET in vitro.Conclusion(s): Lower concentrations of chondroitin sulfate proteoglycan were associated with higher follicular volumes and greater fertilization and ET rates. These associations could merely reflect the maturation of the follicle or a role of the chondroitin sulfate proteoglycan in the fertilization process.     

Comparison of human menopausal gonadotropin administered once and twice daily in an in vitro fertilization and embryo transfer program

This is a retrospective study which evaluates the use of twice-daily (BID) human menopausal gonadotropin (hMG) for follicular stimulation in an in vitro fertilization and embryo transfer (IVF-ET) program from February to June 1985 and compares it to daily (QD) hMG from August to December 1984. All QD patients were begun on 2 ampoules of hMG, and BID patients on 2 ampoules twice daily. Individual patient responses to hMG determined subsequent doses so as to achieve continuously rising estradiol levels. The BID stimulation scheme appears to increase statistically characteristics that would be present in the ideal stimulated cycle such as elevated follicularphase estradiol (E₂) (2125 pg/ml for the BID vs 1581 pg/ml for the QD group) with an increase in the number of patients achieving the desired Jones pattern, increased oocyte retrieval (4.01 vs 2.45), and an increase in the number of transferred concepti (2.69 vs 1.87). No statistical differences were noted in the number of endogenous luteinizing hormone (LH) surges or mean LH. The luteal phase does not appear to have been altered by the frequency of administration. Although the increase in the total (20.0 vs 14.5%) and live-born/ongoing (16.9 vs 11.8%) pregnancy rate per laparoscopy with the BID regimen is not statistically significant, it may be that it is clinically relevant.     

Emotional adaptation following successful in vitro fertilization

OBJECTIVE: To assess the emotional impact of infertility after successful IVF and to compare parents who have undergone IVF (IVF parents) and parents who have not undergone IVF (non-IVF parents) regarding parental stress and the marital relationship during the transition to parenthood. DESIGN: A study with qualitative and longitudinal quantitative assessments. SETTING: University IVF clinics and antenatal clinics in Stockholm. PATIENT(S): Fifty-five IVF mothers, 53 IVF fathers, 40 non-IVF mothers, and 36 non-IVF fathers. INTERVENTION(S): IVF parents were interviewed. All subjects completed self-rating scales in early pregnancy and at 2 and 6 months postpartum. MAIN OUTCOME MEASURE(S): Interviews about perception of infertility and scalar measurement of parental stress and the marital relationship. RESULT(S): Negative feelings related to infertility were not easily overcome among the IVF parents. Their levels of stress related to parenthood were similar to those of non-IVF parents, and both groups reported decreased satisfaction with the marital relationship during the transition to parenthood. CONCLUSION(S): The inability to conceive naturally continues to affect the current lives of a proportion of IVF parents. The results suggest that IVF parents may benefit from counseling with regard to the potential long-term impacts of infertility, disclosure issues, and decisions regarding future children. However, levels of parental stress and patterns of partner satisfaction are similar to those of parents with children conceived "naturally."     

Comprehensive elucidation of amino acid profile in human follicular fluid and plasma of in vitro fertilization patients

Abstract

The assessment of oocyte quality in human in vitro fertilization (IVF) is in focus of intensive studies. Follicular fluid (FF) constitutes the major medium for the developing oocyte and therefore it is a perfect target to search for the biomarkers of female infertility. The objective of this study was to compare the amino acid (AA) profile of human FF and plasma (PL) (from 96 IVF patients) and to examine if the AA composition is related to oocyte quality, IVF results and women’s infertility. In both biological fluids, Gln, Gly and Ala appeared as dominant AAs. Most of AAs are more abundant in PL; the exceptions are Glu, Thr, Ala and Gly, which are higher in FF and Gln, Arg and Phe, the contents of which are similar in both biological fluids. Ser in FF and Met and Phe in PL were detected as potential biomarkers as their content varied depending on the IVF outcome. Significant differences were also detected between the groups of different infertility reasons. Our results suggest that intra-follicular AAs might reflect the condition of the preovulatory follicle and together with PL, AAs can be used to characterize the infertility etiology and oocyte quality related to IVF outcome.     

Protein composition of follicular fluid and oocyte cleavage occurrence in in vitro fertilization (IVF)

The present study was carried out from in vitro fertilization (IVF) attempts to analyze further the total and specific protein contents of 47 follicular fluids yielding one oocyte. The aim was to find correlations between the follicular concentrations of such proteins and the occurrence of coupled oocyte cleavage. These findings would be used as markers of IVF outcome. Two groups of follicular samples were distinguished: one group with cleavage occurrence (25 cases) and another group without cleavage or even fertilization (22 cases). In the group with cleavage a significantly higher level was observed for six proteins: C3 complement fraction and ceruleoplasmin (P <0.02), -antitrypsin and transferrin (P <0.01), and 2-macroglobulin and 2-microglobulin (P <0.001). The data are discussed with respect to changes in follicle permeability with advancing maturity.     

Introduction of blastocyst culture and transfer for all patients in an in vitro fertilization program

Objective: To evaluate the nonselective application of extended embryo culture on the outcome of IVF.

Design: Retrospective analysis.

Setting: Private practice assisted reproductive technology center.

Patient(s): Seven hundred ninety nonselected patients undergoing IVF with controlled ovarian stimulation.

Intervention(s): For day 3 ET, multicell embryos were cultured in human tubal fluid medium and 12% synthetic serum substitute. For day 5 ET, embryos were cultured for 48 hours in S1 medium and then for 48 hours in S2 medium.

Main Outcome Measure(s): Implantation rate (determined by total no. of visualized gestational sacs), ongoing pregnancy rate, and number of embryos available for ET.

Result(s): Respective day 3 and day 5 implantation rates for patients aged <35 years (29.5% and 38.9%), patients aged 35–39 years (20.7% and 28.2%), and all patients combined (23.3% and 32.4%) were statistically significantly different. Significantly more embryos were transferred on day 3 than on day 5 for patients aged <35 years (2.9 vs 2.4), patients aged 35–39 years (3.1 vs 2.6), and all patients combined (3.0 vs 2.5). The difference in ongoing pregnancy rates per retrieval was statistically significant for day 3 compared with day 5 transfers for all patients combined (35.9% vs 43.8%). Cancellation rates for transfer after retrieval increased significantly for day 3 compared with day 5 transfer (2.9% vs 6.7%).

Conclusion(s): These results demonstrate the feasibility of using extended embryo culture in a nonselective manner for couples undergoing IVF. Overall, extended embryo culture was associated with a significant increase in pregnancy rates and implantation rates and a significant decrease in the number of embryos transferred. The rate of multiple implantation among patients aged <35 years warrants consideration of single blastocyst transfers for this group.     

Chromosome investigation in in vitro fertilization failure

Purpose A chromosomal complement of 227 human oocytes was studied to provide information on the frequency and type of chromosomal abnormalities in oocytes failing in vitro fertilization.Results Normal haploid chromosome complement was found in 54.6%; chromosomal abnormalities consisting of diploid sets were identified in 16.7% and aneuploidy was observed in 26%. Premature condensation of sperm chromosomes of the G1-phase was observed in 22.9% oocytes. Male infertility was correlated with an increase in the rate of aneuploidy when compared with tubal infertility. The rate of chromosome abnormalities for the oocytes recovered from women who had no fertilized oocytes was significantly higher compared to those with at least one oocyte fertilized.Conclusion A high frequency of chromosome abnormalities in unfertilized oocytes suggests that natural selection against chromosome abnormalities may occur even prior to fertilization.     

Viscosity and refractive index of follicular fluid in relation to in vitro fertilization

Purpose: To set the standard values of follicular fluid viscosity and refractive index, and to investigate a possible relationship between these physiological parameters and the outcome of in vitro fertilization treatment. Design and Results: 128 samples of follicular fluid were collected from 40 in vitro fertilization patients. Viscosity determinations (centipoise; mean ± SD) for shear rates of 23, 46, 115, and 230 were 2.04±0.86, 1.84±0.49, 1.48±0.27, and 1.38±0.22, respectively. The average (± SD) refractive index was 1.030±0.002. There was no significant difference between the values of thawed frozen fluids and fresh samples of the same specimens. The data showed no correlation between follicular fluid viscosity or refractive index and the presence of oocytes, their maturation grade or their fertilizing capacity. Conclusions: For the first time, values of the viscosity and refractive index of follicular fluid obtained during in vitro fertilization have been determined. However, these preliminary results did not reveal any relationship between the physiological parameters examined and the outcome of in vitro fertilization treatment.     

Beta-Endorphin levels in the preovulatory follicles and the outcome of in vitro fertilization

The level of immunoreactive beta-endorphin-like (IR--EP) material(s) was determined in follicular fluid samples obtained from patients treated for in vitro fertilization (IVF). Follicles containing morphologically mature oocytes had slightly lower concentrations of IR--EP. The total IR--EP content was similar in follicles with oocytes at different degrees of maturity. A smaller amount of IR--EP in the follicle fluid was associated with a greater developmental potential of the oocytes, which formed embryos that reached the three- to six-cell stage at 40–44 hr after insemination (r=–0.35, P<0.026). A higher IR--EP content was seen in fluid samples containing oocytes that remained uncleaved after fertilization (P<0.05 by Duncan's multiple-range test).     

Hormonal stimulation for in vitro fertilization: A comparison of fertilization rates and cytogenetic findings in unfertilized oocytes

Objective The purpose of this study was to compare fertilization and aneuploidy rates after two stimulation protocols in an IVF program.Design This was a retrospective study.Setting The study took place in the IVF laboratory of an Infertility Department.Methods In 349 treatment cycles, clomiphene citrate (CC) and human menopausal gonadotropin (hMG) were used in one group (N =233) and hMG after treatment with a gonadotropin-releasing hormone agonist (GnRHa) in two other groups (long protocol): goserelin (N =73) and buserelin (N =43). Cytogenetic analysis was performed on all unfertilized oocytes in both groups. Results Fertilization rates were significantly higher in the GnRHa/hMG group than in the CC/hMG group, but cleavage rates and embryo quality were not different. Of 736 oocytes prepared for cytogenetic analysis, 256 were karyotyped: 172 were found to be euploid and 84 aneuploid. More oocytes were aneuploid in the GnRHa/hMG group than in the CC/hMG group and this difference was statistically different after analysis of the data using a specially designed mathematical model.Conclusion If no selection against chromosomally abnormal oocytes takes place at the time of fertilization, more abnormal oocytes are harvested with GnRHa/hMG protocols than with CC/hMG. If, on the other hand, there is a selection against oocytes with some chromosomal imbalance, there is no intrinsic effect of GnRH agonists on the chromosomal complement of the oocyte, and the real aneuploidy frequency in all oocytes, fertilized and unfertilized, is the same in the GnRHa/hMG and in the CC/hMG group.     

Outcome of patients with one ovary in an in vitro fertilization program

To determine whether the absence of one ovary would influence adversely the outcome of in vitro fertilization, the results of 60 laparoscopic oocyte retrieval procedures in 34 single-ovary patients were compared with the outcome for all laparoscopic retrieval cycles during the same period (January 1984–August 1986) in patients with tubal infertility and two accessible ovaries (559 cycles in 335 patients). The median age was younger in the one-ovary group (31 vs 32) (P < 0.05). The group with two accessible ovaries had significantly more follicles aspirated (median, 7 vs 5; P < 0.001) and more oocytes obtained (median, 5 vs 4; P < 0.001) per retrieval compared with the single-ovary group. However, the single-ovary group had a higher percentage fertilization (69 vs 62%; P < 0.05) so that the number of embryos transferred per transfer patient (median, 3 vs 3) and pregnancy rate per cycle (9.7 vs 15.6%) did not differ significantly between groups. We conclude that patients with a single ovary have a compensatory increase in the ability to produce oocytes which may have greater potential for fertilization and subsequent development.     

The day of initiation of human menopausal gonadotropin stimulation affects follicular growth in in vitro fertilization cycles

The attainment of synchronous follicular development in human menopausal gonadotropin/human chorionic gonadotropin-stimulated cycles for in vitro fertilization (IVF) continues to be a perplexing problem. Two regimens of follicle stimulation for IVF cycles were, therefore, compared. Twenty-nine patients commenced human menopausal gonadotropin (hMG) therapy on day I of the menstrual cycle (Group I), while 30 women received hMG from the third day of the cycle (Group II). The hMG therapy was tailored to the individual patients's response, based on ultrasonographic measurements of follicular size and serum estradiol (E₂) levels. Both groups of patients received a mean of 19.6±1.4 ampules of hMG over a mean of 6.1±0.2 days. The pattern of serum E₂ and progesterone levels in the periovulatory and luteal phase was not affected by the day of initiation of hMG therapy, although Group I patients demonstrated lower (P<0.05) E₂ levels on the 2 days prior to human chorionic gonadotropin (hCG) administration. In terms of follicle growth, Group II follicles consistently demonstrated a significantly (P<0.01,x ² test) larger proportion of medium- and large-sized follicles compared to Group I follicles on almost all of the days when ultrasonographic measurements were taken. In addition. Group II follicles demonstrated an earlier shift (day—1) to the larger follicles than Group I follicles (day 0). Significantly (P<0.001) more oocytes were recovered per uspirated follicle in Group II patients, but the fertilization rate per oocyte was greater (P<0.003) for Group I oocytes. Nevertheless, pregnancy rates did not differ between the two groups. It is suggested that a difference between the two groups of patients in the quantity or quality of gonadotropin receptor sites in the early part of the follicular phase may account for both the diminished E₂ production in the follicular phase and the persistent depressed follicular growth in Group I patients.     

Levels of oxytocin-like activity and progesterone in follicular fluid from in vitro fertilization cycles

Oxytocinlike immunoreactivity, estradiol, and progesterone were measured in follicular fluid collected during oocyte collection in an in vitro fertilization program in which clomiphene citrate was used to stimulate follicular development. Follicles which yielded morphologically normal embryos after fertilization of the oocyte had oxytocin concentrations ranging from <10 to 600 ng/liter. Oxytocin concentrations did not differ between follicles from 12 pregnancy cycles (median, 169; N=21 and follicles from 12 nonpregnancy cycles (median, 110; N= 18). Oxytocin concentrations were correlated negatively with progesterone concentrations (Spearman's rank correlation coefficient r=–0.50; P=0.001). In cycles with some follicles having progesterone concentrations <10 and some >10 mol/liter, oxytocin concentrations were higher in the less progestogenic follicles in 15 of 16 cases.     

Intrafollicular and serum levels of leptin during in vitro fertilization cycles: comparison between the effects of recombinant follicle-stimulating hormones and human menopausal gonadotrophin

Objectives.?To compare the effect of recombinant follicle-stimulating hormones (r-FSH) and human menopausal gonadotrophin (hMG) on leptin levels in serum and follicular fluid (FF) during in vitro fertilization IVF/ET treatment, and to investigate whether leptin levels in the follicular fluid and/or serum are correlated with IVF success.

Methods.?Sixty-three patients undergoing IVF cycle were subdivided into two groups. r-FSH was used to for controlled ovarian hyperstimulation in 29 patients (Group A) while, hMG was used in 34 patients (Group B). Our main outcomes were serum and FF leptin on the day of oocyte collection.

Result(s).?The two groups were comparable in age, body mass index (BMI), indications for IVF/ET, E2 level on human chorionic gonadotrophin day, number of retrieved oocytes, fertilization rate, number of transferred embryos and pregnancy rate. Serum and FF leptin levels were similar between the two study groups. Additionally, no correlation was found between levels of leptin in either serum or FF and cycle results such as: number of retrieved oocytes, fertilization rate and pregnancy rate.

Conclusions.?r-FSH and hMG have been found to have comparable effects on leptin levels in the serum and the FF of patients undergoing IVF/ET. Additionally, leptin levels in both serum and FF on day of retrieval have no correlation to IVF/ET outcome.