Increased human menopausal gonadotropin dose during the early follicular phase: Effect on follicular recruitment and treatment outcome

Summary To investigate whether the provision of increased gonadotropins would enhance follicular recruitment and selection, women not responding (N=18) to our standard clomiphene citrate (CC)-human menopausal gonadotropin (hMG) regimen were treated with increased hMG (treatment cycle II). Estradiol levels were higher in treatment cycle II and these differences were significant on days 8 and 9 (P0.05 andP0.03, respectively). On day 9, better follicular development was seen in cycle II (P0.05). While none of the patients responded in cycle I, 10 of 18 responded in cycle II. Of the 10 responders, 2 conceived following in vitro fertilization and embryo placement. Increasing the dose of hMG improved the development of a cohort of follicles so that aspiration and possible pregnancy were achieved in women who were previously unresponsive to therapy.     

Routine laser assisted hatching results in significantly increased clincal pregnancies

Purpose: To elucidate the appropriateness of current indications for assisted hatching (AH) in cleavage stage human embryos and to confirm our preliminary findings that only young patients (about 67%) benefit from AH. Methods: Prior to transfer, 2 of 3 embryos selected for ET were subjected to laser assisted hatching (LAH). Control group consisted of patients matched by similar characteristics and protocol except LAH was not performed. Results: The clinical pregnancy rate in women 36 years was 64.9% (24/37) for embryos subjected to LAH but was significantly lower (p = 0.029) in the control (33.3%; 10/30). The implantation rate in women 36 years in the test group was 38.1% (40/105) that was significantly higher than that of the control group (17.5%, 14/80; p = 0.0039). Conclusions: LAH is beneficial for women 36 years but not for women 37 years, for embryos with thin zonae (16) but not with thick zonae (17), and for those with repeated failures (37–50%).     

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.     

Spermatogenic patterns and early embryo development after intracytoplasmic sperm injection in severe oligoasthenozoospermia

Purpose: Evaluate the influence of different baseline spermatogenic patterns [meiotic pattern (normal or abnormal), sperm concentration ( > 1 × 10⁶/mL or 1 × 10⁶/mL), and the combined meiosis–sperm concentration pattern] on early embryo development in severe oligoasthenozoospermia. Methods: Embryo outcomes (fertilization rate, cleavage rate, and 4-cell stage embryo division rate on day 2) after IVF–ICSI in 75 oligoasthenozoospermia and 79 normozoospermic males. Results: The embryo division rate was significantly lower in oligoasthenozoospermia compared to normozoospermia (50.43% vs. 58.72%, p < 0.01) and in the oligoasthenozoospermia group for meiotic anomalies (43.40%), sperm concentration 1 × 10⁶/mL (44.35%), and the combined pattern 1 × 10⁶/mL with meiotic anomalies (37.17%). Logistic regression analysis showed a synergic effect (OR = 2.00; 95% CI = 1.28–3.12) when the two spermatogenic patterns predictive of slow embryo development [meiotic anomalies (OR = 1.49; 95% CI = 1.03–2.15) and sperm concentration 1 × 10⁶/mL (OR = 1.53; 95% CI = 1.09–2.13)] were present. Conclusions: The data suggest that the early embryonic developmental capacity is inversely related to the severity of spermatogenic impairment (meiotic anomalies and/or sperm concentration 1 × 10⁶/mL).     

Spectrophotometric analysis of human follicular fluid with regard to in vitro fertilization (IVF) parameters,follicular protein,and hormone content

Purpose Our purpose was to investigate possible relationships with spectrophotometric absorbance (458-nm region) and biochemical variables in follicular fluid (FF) as well as in vitro fertilization (IVF) outcome.Methods This study included 227 normal ovulatory women undergoing oocyte retrieval for IVF. Blooduncontaminated fluid samples, identified by spectrophotometry, were investigated. Spectrophotometric absorbance of FF at 458 nm (n = 426), as well as hLH, FSH, PRL, hCG, testosterone, sialic acid, ₁-antitrypsin and plasminogen of selected fluids, was analyzed.Results Small-volume follicles (2 ml) were associated with higher absorbance profiles (P <0.05), when compared to volumes greater than 2 ml. Our data suggest that the presence or absence of an oocyte, the potential of an oocyte to fertilize or cleave, failed to show any relationship with maximum FF absorbance at 458 nm. Maximum absorbances were significantly lower in FF from patients who subsequently became clinically pregnant (P =0.039). No correlation between FF absorbances and biochemical parameters (P >0.15) were established. Conclusions Absorbance of clear FF at 458 nm should not be viewed as the single parameter to predict oocyte development in vitro.     

The relation between depth of trophoblastic invasion and β-HCG levels in tubal pregnancies

-HCG (human chorionic gonadotropin) values of over 2500 I.U./l are associated with higher failure rates for therapy with prostaglandin F2 alpha in tubal pregnancies. The purpose of our study was to ascertain if the 2500 I.U./l limit correlates with histopathology. We therefore compared the pre-operative -HCG-values and intraluminal and extraluminal trophoblast growth in tubal pregnancy. Purely intraluminal trophoblast was significantly more frequent in patients of group I (-HCG < 2500 I.U./1), while group II patients (-HCG > 2500 I.U./l.) almost exclusively had extraluminal growth (P=0.0045). Since the efficacy of prostaglandin F2 alpha therapy depends on intact tubal musculature the correlation of the -HCG threshold level with histopathologic findings may explain the high failure rate in patients with -HCG values above 2500 I.U./l. Correspondence to: M. Klein     

Antral follicle counting in predicting the retrieved oocyte number after ovarian hyperstimulation

Purpose: To evaluate the antral follicle (AF) counting in predicting the outcome after controlled ovarian hyperstimulation (COH) and IVF-ET.Methods: Infertile women who accepted the COH and IVF-ET were included prospectively. Day-3 AF number was determined by transvaginal sonography. They were divided into three groups: Group 1: 3 AF; Group 2: 4–10 AF; Group 3: 11 AF. Retrieval oocyte number, embryo number, pregnancy rate (PR), and implantation rate (IR) of the three groups were compared.Results: A total of 372 cycles were included. Patients in Groups 1, 2, and 3 were statistically different in age (35.3 vs. 31.9 vs. 28.5), Day-3 FSH (14.3 vs. 5.9 vs. 4.1), cancellation (34.4% vs. 2.7% vs. 0.9%), gonadotropin dosage, retrieval oocyte number (2.9 vs. 8.2 vs. 14.5), embryo number (2.2 vs. 6.5 vs. 11.7), PR (11.1% vs. 34.6% vs. 35.0%), and IR (3.0% vs. 8.2% vs. 8.9%).Conclusions: Patients with 3 AF have a significantly higher cancellation rate, fewer retrieved oocyte number, and lower PR. Combination of AF counting and basal FSH level increased the sensitivity in predicting the ovarian reserve. Retrieved oocyte number could be predicted by the formula: oocyte = 0.802 × AF + 2.01.     

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.     

Effect of Intracellular Ca2+ Chelation with the Acetoxymethyl Ester-Derived Form of Bis(o-Aminophenoxy)Ethane-N,N,N,N′,N′-Tetraacetic Acid on Meiotic Division and Chromosomal Segregation in Mouse Oocytes

Purpose: Our purpose was to ascertain the effect of intracellular Ca ²⁺ chelation on the chromosomal distribution and segregation of mouse oocytes during maturation in vitro. Methods: Germinal vesicle oocytes were loaded with the acetoxymethyl ester-derived form of bis(o-aminophenoxy)-ethane-N,N,N,N-tetraacetic acid (BAPTA-AM). Chromosomal distribution and segregation of control and BAPTA-AM-treated metaphase II (MII) oocytes were evaluated at 16 hr, and intracellular ATP content at 0, 1, and 16 hr after BAPTA-AM loading. Results: BAPTA-AM treatment decreased (P 0.05) the potential for in vitro maturation, increased (P 0.0001) the percentage of oocytes displaying an abnormal distribution of metaphase II chromosomes in the meiosis II spindle and aneuploidy, and decreased (P 0.005) the ATP content at 0, 1, and 16 hr of culture compared to the control groups. Conclusions: These findings raise some concern about any other condition/drug that may directly or indirectly decrease the intracellular Ca ²⁺ concentration in human oocytes.     

Serum levels of zinc and copper in hydatidiform mole

Objective The aim of this study was to identify any association between serum levels of trace elements zinc and copper and development of hydatidiform mole.Study design Blood samples of 35 patients with complete hydatidiform mole and 34 pregnant women controls were evaluated for serum levels of zinc and copper. Students t-test was used for comparison of mean values of serum zinc and copper and for demographic variables. p0.05 was considered statistically significant.Results Levels of zinc in serum were found to be significantly higher in hydatidiform mole patients than controls (86.3 vs. 68.7 g/l, p=0.027). Serum copper levels were significantly lower in hydatidiform mole patients than controls (123.8 vs. 147.8 g/l, p=0.039; Table 2).Conclusion This is the first study in the English scientific literature reporting higher serum levels of zinc and lower serum levels of copper associated with complete hydatidiform mole.     

Interleukin-1Β, interleukin-6, and growth hormone levels in human follicular fluid

Purpose To investigate possible relationships of interleukin-1 (IL-1, interleukin-6 (IL-6), and growth hormone (GH) with biochemical variables in human follicular fluid (FF) and selected in vitro fertilization (IVF) parameters.Methods A total of 67 FF samples (n=67 patients undergoing oocyte retrieval for IVF) was evaluated. IL-1, IL-6, GH, hLH, FSH, PRL, hCG, testosterone, total protein, fibrinogen, sialic acid, ₁-antitrypsin, plasminogen levels, and spectrophotometric absorbance at 458 nm were analyzed for selected FF. IL-6 and GH levels of serum and FF samples were also compared (n=23).Results Immunoreactive levels of IL-1, IL-6, and GH were detected in all FF samples. A positive correlation existed for IL-6 (r=0.5069, P=0.0161 when serum-to-FF levels were compared (concentration ratio, 11.857). Smaller-volume follicles (<4 ml) were associated with high IL-1 levels (P=0.0229, and an additional tendency of IL-1 to decrease with increasing embryo cleavage and scoring was observed. With the exception of a weak positive correlation between follicular IL-1 and testosterone levels (r=0.3128, P=0.025, no other relationship with biochemical variables or IVF parameters (etiology, e.g., endometriosis) could be implicated.Conclusions Substantially higher IL-6 levels occurred in FF compared to serum, thus supporting intrafollicular production. Interleukin- 1,IL-6, and GH levels in FF are, however, unsuitable markers for in vitro fertilization outcome.     

The association between chlamydial-specific IgG and IgA antibodies and pregnancy outcome in an in vitro fertilization program

Chammydial-specfic IgG and IgA antibodies were determined by a single serovar (L2) immunoperoxidase assay (IPA) in the serum of all patients that have conceived in an in vitro fertilization and embryo transfer (IVF & ET) progrom (n=106) and in a group of patients that went through the program at the same period of time and did not conceive (n=94). The prevalence rate of elevated IPA IgG (titers1128) and IPA IgA (titers116) specific to chlamydiae was significantly higher (P<0.001) in the IVF&ET pregnancy loss and nonconception groups (failures) versus the IVF&ET term pregnancy group (successes) (74 vs 47%, odds ratio=4.1, and 34 vs 14%, odds ratio=4.3, respectively). Stepwise discriminant analysis revealed that elevated specific chlamydial IgG had the greatest effect on the variance between successes and failures in this study group. Our study indicates the possible role of past or chronic active chlamydiae infection on the take-home baby rate in an IVF&ET program.     

Assessment of the Dominant Abnormal Form Is Useful for Predicting the Outcome of Intracytoplasmic Sperm Injection in the Case of Severe Teratozoospermia

Purpose: Our purpose was to investigate the relation between the dominant sperm anomaly and sperm morphology and the outcome of intracytoplasmic sperm injection (ICSI). Methods: Two hundred ninety-five patients who underwent a total of 181 cycles of in vitro fertilization (n = 168) and/or 217 cycles of ICSI (n = 177) between July 1995 and May 1997 at Keio University Hospital were investigated. Results: The rates of fertilization and pregnancy were 63.3 and 27.8%, respectively, in ICSI cycles with 4% normal forms. When the percentage of strictly normal morphology was 4, the fertilization rate was lower in the case of severely tapered head (13.0%; n = 4) than in the cases of other deformities in ICSI. The acrosomal defect made no difference in the fertilization rate with ICSI. Conclusions: The predominant abnormal form affects the ICSI outcome in the case of 4% normal forms.     

Identification of follicles with fertilizable oocytes by sequential ultrasound measurements during follicular development

Patients undergoing ovulation induction for in vitro fertilization and embryo transfer (IVF-ET) were monitored daily with serum estradiol-17 (E₂) and ultrasound. The location of each individual follicle was established by taking ultrasound images through serial sections of the ovary. The diameter of each follicle and the volume of its follicular wall (FW) were determined from ultrasound images using a computer-controlled image analyzer. A total of 44 follicles from nine patients was studied, with an overall fertilization rate of 46%. In all patients, serum E₂ levels increased prior to human chorionic gonadotropin (hCG). Whereas changes in either the average diameter or the volume of the entire follicle did not identify follicles with fertilizable oocytes, FW volume measurements were predictive. Prior to hCG, FW volume increased 24±8%/day in follicles with fertilizable oocytes but decreased 3±6%/day in follicles with nonfertilizable oocytes (P<0.05). Three major patterns of follicular development were observed for follicles with nonfertilizable oocytes: slow growing (<20% increase in FW volume), nongrowing (no change in the FW volume), and degenerating (a decrease in the FW volume), suggesting that these follicles are postmature. These data demonstrate that FW volume measurements made from sequential ultrasound images provide an accurate method to identify those follicles that contain fertilizable oocytes.Presented in part at the VI World Congress on in Vitro Fertilization and Alternate Assisted Reproduction, Jerusalem, Israel, April 1989.     

Identifying the Public’s Knowledge and Intention to Use Human Cloning in Greek Urban Areas

Purpose: The understanding of the publics knowledge on human cloning (HC) and its acceptability are considered important for the development of evidence-based policy making. The aim of this research study was to investigate the demographic and socioeconomic variables that affect the publics knowledge and intention to use HC in urban areas of Greece. Additionally, the possible association of religiousness with the knowledge and the intention to use HC were also investigated.Methods Individual interviews were conducted with 1020 men and women of urban areas in Greece. Stratified random sampling was performed to select the respondents. Several scientists, experts in HC, evaluated the content of the instrument initially developed. The final questionnaire was consequently the result of a pilot study.Results Almost half of the respondents (51.5%) believed that HC is a sort of in vitro fertilization and 42.9% that it has already been applied to human being. They were not aware that the cloned fetus grows in the womans uterus (41.5%) and that HC could regenerate human organs (41.7%). The acceptability of human cloning for the cure of terminal diseases and transplantation need is very high (70.7% and 58.6%, respectively). The publics intention to have recourse to cloning on the grounds of bringing back to life a loved person or because of reproductive disorders was reported as desire by 35% and 32.5%, respectively. The occupational category (scientists, self-employed, and artists), the Intention to use HC, and the number of children are highly significant predictors of valid knowledge about HC. Low rates of church attendance appeared to relate with high reported Intention to use HC, and increasing scores of valid knowledge about HC increased the publics Intention to use HC.Conclusions A number of specific demographic and socioeconomic characteristics and high scores of knowledge provide a persuasive justification in demonstrating intention toward HC. The current study suggests that these findings should receive further attention by policymakers and scientists within the Greek context.     

ANDROLOGY: Relationships Between Sperm Motility Characteristics Assessed by the Computer-Aided Sperm Analysis (CASA) and Fertilization Rates In Vitro

Purpose: Some studies have suggested that computer-aided sperm analysis (CASA) estimates of concentration and movement characteristics of progressively motile spermatozoa are related to fertilization rates in vitro. However, it has also been suggested that the greater number of motility parameters assessed by CASA does not imply more precision in predicting fertility. This study was carried out to investigate the relationships between the CASA estimates and fertilization rates in vitro.Methods: Semen quality analysis was performed using CASA in 136 in vitro fertilization-embryo transfer (IVF-ET) cycles with at least 3 oocytes collected. The CASA estimates before and after swim-up were compared between 108 cycles with fertilization rate >50% (good group) and 28 cycles with fertilization rate 50% (poor group).Results: Before swim-up, there were significant correlations between fertilization rates and CASA estimates, including amplitude of lateral head displacement (ALH) (r = .269), curvilinear velocity (VCL) (r = .297), straight line velocity (VSL) (r = .266), and rapid sprm movement (Rapid) (r = .243). There was also a significant correlation between the fertilization rates and straightness (STR) after swim-up (r = –0.178). As for sperm movement characteristics, there were significant differences of ALH (p < .005), VCL (p < .001), VSL (p < .005), and Rapid (p < .01) between good and poor groups before swim-up. After swim-up, there were significant differences of VCL (p < .005), average path velocity (VAP) (p < .005), and Rapid (p < .05) between the two groups.Conclusions: These results indicate that some of the CASA estimates provide reliable estimation of the fertilizing ability of human sperm. There were significant differences of the two sperm movement characteristics, including VCL and Rapid (before and after swim-up), indicating that the total distance traveled by rapid sperm movement might be important in human sperm fertilizing abilities.     

Zona Thinning with Noncontact Diode Laser in Patients Aged ≤37 Years with No Previous Failure of Implantation: A Prospective Randomized Study

Purpose: Zona thinning (ZT) is a technique used to improve pregnancy rates among patients 38 years old and/or patients presenting previous implantation failure. The objective of the study was to determine whether ZT has a beneficial effect on patients younger than 37 years who are undergoing the first ICSI attempt. Methods: A total of 103 patients submitted to ICSI for the first time and those aged 37 years were divided in a prospective and randomized manner into two groups: group I, patients submitted to ZT (n = 51) (a laser diode with 1.48-m wavelength (Fertilaser) was used for the procedure); group II, patients with no ZT (n = 52). In both groups, embryo transfer was performed on the second day. Results: The age of group I patients (31.8 ± 3.6) did not differ (P = 0.53) from that of group II patients (31.4 ± 3.6). The number of metaphase II oocytes was similar(P = 0.76) for the two groups (group I = 9.12 ± 5.27; group II = 8.67 ± 5.02). The average number of embryos available per transfer of group I (6.14 ± 4.02) did not differ (P = 0.69) from that of group II (5.75 ± 3.83). The number of embryos transferred was similar (P = 0.61) for the two groups (group I = 2.76 ± 0.9; group II = 2.87 ± 0.79). The thickness of the zona pellucida of group I embryos (16.6 ± 2.2 m) did not differ (P = 0.08) from that of group II embryos (17.1 ± 1.7 m). The rate of embryo implantation (20.8%) and the rate of clinical pregnancy per embryo transfer (40.3%) were higher for group II than for group I (17.7% and 33.3%, respectively), but the difference was not significant (P = 0.55 and P = 0.54). Conclusions: These results suggest that ZT in the population aged 37 years and with no previous failure of implantation may have no impact on intracytoplasmic sperm injection success rates.     

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.     

First trimester diagnosis of partial mole

Background Partial mole is one of the two distinctive subtypes of hydatidiform mole. It is usually paternally derived triploid conceptions in which embryonal development occurs in association with trophoblastic hyperplasia. The definite diagnosis is confirmed by pathological and cytogenetic studies. Ultrasound might be helpful to diagnose partial mole in the first trimester.Case A 25-year-old woman, gravida 2, para 0-0-1-0, was initially seen for antenatal care at 6 weeks pregnant. Ultrasound was undertaken at 13 weeks pregnancy due to her first fetal anomaly, which demonstrated partial mole and embryonic death. The serum hCG was 190,900 mIU/ml. Suction curettage was performed without complication. Histopathological study confirmed partial mole and cytogenetic study of the placenta revealed an uncommon karyotype, mosaicism of triploid (69,XXX/69,XXY). Serum hCG was declined and negative at 8 weeks. The patient was well and serum hCG remained normal throughout 6 months of follow-up.Conclusion Although the majority of partial mole pregnancies cannot be detected by routine first trimester ultrasound examination, first trimester ultrasound can be helpful in some cases, such as this one. If partial mole is sonographically suspected, it should be confirmed with histopathology and cytogenetic studies. The management is similar to complete mole including prompt evacuation and careful monitoring of hCG.     

Characterization and prediction of IVF cycles generating “slow-cleaving” embryos

Purpose To characterize and predict cycles generating slowcleaving embryos in in vitro fertilization, 86 cycles were retrospectively divided into two groups (slow, n=41, and fast, n=45 according to whether the number of blastomeres per embryo on day 3 was or > than the mean of the distribution, respectively.Results Cycles generating slowcleaving embryos were treated with luteinizing hormonereleasing hormone agonist before ovarian stimulation for a shorter period (12.1±0.5 versus 15.6±1.1 days; P0.01) and had higher immaturity grade of oocyte-corona-cumulus complexes which resulted in embryos (1.6±0.1 vs 1.3±0.1; P0.05) when compared to cycles producing fastcleaving embryos. Both variables entered in a logistic regression model applied in order to predict the probability of a cycle generating slowcleaving embryos (goodness-of-fit chisquare=180.0, degrees of freedom (df)=80, P=0.4786. This model predicted correctly 86.7% (13 of 15) of cycles generating slowcleaving embryos and 83.3% (10 of 12) of cycles producing fastcleaving embryos when the estimated probability of a cycle producing slowcleaving embryos was 0.7 or 0.3, respectively.Conclusion Shorter treatment with hormone-releasing hormone agonist before ovarian stimulation and higher immaturity grade of oocyte-corona-cumulus complexes which result in embryos are predictive characteristics of in vitro fertilization cycles generating slow-cleaving embryos.