First birth following assisted sperm fusion insemination using sperm bound to zona pellucida.

PURPOSE: To report a live birth after transfer of a vitrified-warmed blastocyst produced by assisted sperm fusion insemination (ASFI). METHODS: Oocyte retrieval and in vitro fertilization (IVF) were performed on a 37-year-old woman. Six hours after IVF, an oocyte exhibited a single polar body and so was defined as an unfertilized oocyte. A motile sperm was collected from the zona pellucida of the unfertilized oocyte by an injection needle. The motile sperm was pressed onto the membrane of the unfertilized oocyte. RESULTS: Two oocytes were matured and subjected to IVF. One of the 2 oocytes exhibited only one polar body and was defined as an unfertilized oocyte at 6 h after IVF; this oocyte then was subjected to ASFI. Two pronuclei were observed on the next day and cultured to the blastocyst stage. This embryo achieved blastocyst status and was vitrified on day 5. The resulting vitrified-warmed blastocyst was transferred, resulting in pregnancy and subsequent delivery of a healthy boy. CONCLUSION: This report describes the first case of a successful birth following transfer of a vitrified-warmed blastocyst produced by ASFI.

Difference in the size of the placenta and umbilical cord between women with natural pregnancy and those with IVF pregnancy.

PURPOSE: The purpose of this study was to compare the sizes of the placenta and umbilical cord in women with natural pregnancy versus those undergoing in vitro fertilization (IVF). METHODS: Overall, 1610 cases of uncomplicated single pregnancies with vaginal delivery at ≥ 37 weeks of gestation were included in this study. The patients were divided into two groups: natural pregnancy group (n = 1453) and IVF pregnancy not including intracytoplasmic sperm injection (ICSI) treatment (n = 157). The groups were compared in terms of gestational week, maternal age, parity, maternal weight gain, prepregnancy maternal BMI, infant weight at birth, infant head circumference, placental weight, cross section of the placenta, cross section of the umbilical cord, insertion site of the umbilical cord, and umbilical cord length. Stepwise selection and multivariate logistic regression were used for statistical analysis to correct the result as an independent factor. RESULTS: There was no difference in the size of the placenta and umbilical cord between women with natural pregnancy and with IVF, but the incidence of velamentous insertion of the cord was significantly increased in women with IVF pregnancy (adjusted odd ratio [AOR] 1.72, 95% confidence interval [CI] 1.08-2.72, p = 0.026). CONCLUSIONS: Although there is no difference in placental weight and cord size, velamentous insertion of the umbilical cord increases in IVF pregnancy and needs careful observation during the delivery process.

Morphologically poor blastocysts could affect the implantation rate of a morphologically good blastocyst during a double-blastocyst transfer for patients who have experienced repeated implantation failures.

PURPOSE: Increasing the number of transferred blastocysts sometimes is selected for patients with repeated implantation failure (RIF). To confirm this strategy, the pregnancy rates (PRs) were compared among the groups who had transferred either a single morphologically good blastocyst (MGB group), double blastocysts with both a MGB and a morphologically poor blastocyst (MGB + MPB group), or a double-BT with 2 MGBs (two-MGB group). METHODS: This study was performed between April, 2009 and September, 2014, including 634 cycles for 354 patients with RIF. All the patients received cryopreserved blastocysts in either hormone replacement or natural ovulatory cycles. The included MGBs were at more than the Gardner grade 3BB stage. The PR and implantation rates (IRs) among the three groups were statistically evaluated by the chi-square test. Statistical significance was set at P < .01. RESULTS: Although the PRs were similar in these three groups, the IR in the MGB + MPB group was significantly lower than that of the MGB group. The rate in the two-MGB group also was significantly lower than that of the MGB group. CONCLUSION: A double-BT with a MGB and a MPB does not increase the pregnancy rate, compared with a single-BT with a MGB among patients with RIF.

Dichorionic triplets following frozen-thawed poor-stage embryo transfer: a report of two cases and a review.

BACKGROUND: We describe two cases of dichorionic triplet pregnancy after a frozen-thawed poor-stage embryo transfer. A 39-year-old and a 41-year-old woman underwent ART treatment. The first patient underwent intracytoplasmic sperm injection (ICSI) at 34 years of age, and two frozen-thawed poor-stage embryos were transferred at 39 years of age with assisted hatching, resulting in a trichorionic triamniotic triplet pregnancy. The second patient underwent ICSI, and two poor-grade blastocysts were transferred followed by assisted hatching, resulting in a dichorionic triamniotic triplet pregnancy. In the first case, the heartbeat of one monozygotic twin fetus had stopped on day 48 post-transfer (9 weeks 2 days), resulting in a dichorionic diamniotic twin pregnancy. A healthy boy and girl were delivered by elective caesarean section at 36 weeks, 5-days gestation. In the second case, the patient underwent selective reduction of the monochorionic twins, resulting in a single pregnancy that was vaginally delivered without any problems at 38 weeks 0-days gestation. SHORT CONCLUSIONS: Numerous factors may be associated with the development of a monochorionic pregnancy; however, controversies still remain. The present morphological grading for embryos is insufficient for inhibiting the development of a monochorionic pregnancy.

An abdominal ectopic pregnancy following a frozen-thawed ART cycle: a case report and review of the literature.

BACKGROUND: Ectopic pregnancy (EP) occurs in 1% of pregnancies and is reported to be more common in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) pregnancies. An abdominal ectopic pregnancy (AEP) is a rare form of EP, and there are few reports of an AEP after IVF/ICSI. In this case report, a rare case of AEP after frozen-thawed cycle of ICSI is presented. CASE PRESENTATION: After a frozen-thawed cycle of ICSI, the beta-human chorionic gonadotropin (HCG) level at 4 weeks 0 days of gestation was 3.4 IU/L. Subsequent dysfunctional uterine bleeding was mistaken for menstruation; however, an AEP of 9 weeks with a fetal heart beat was observed by ultrasound. After the AEP was observed by ultrasound, it was extracted laparoscopically. CONCLUSION: A rare case of an AEP, which developed after frozen-thawed cycle of ICSI, presented with a very low serum HCG level. Even if the HCG titer is low, follow-up HCG levels and frequent medical examinations are necessary.

A comparison of the clinical outcomes of embryos derived from intracytoplasmic sperm injection after early fertilization check and conventional insemination using sibling oocytes.

PURPOSE: In order to identify the real contribution of early fertilization check (EFC) for reproductive outcome, we compared the developmental potential of embryos derived from intracytoplasmic sperm injection (ICSI) after EFS with those from conventional insemination in sibling oocytes. METHODS: Between April 2009 and April 2012, a total of 3249 oocytes in 386 patients were recruited following conventional insemination. Oocytes showing a second polar body (2ndPB) after an EFC were considered to be fertilized oocytes (IVF group), but, oocytes not showing a 2ndPB after EFC were placed into the ICSI group. The incidence of morphologically good embryos (MGE) on day 3, the blastocyst formation (BL), and the development of full blastocysts (full-BL) on day 5 were compared between the two groups. The clinical pregnancy rate was compared between the cycles with only conventional insemination or ICSI after EFC of the embryos being transferred. RESULTS: The fertilization rates in both the IVF and the ICSI groups were 48.1 and 73.9 %, respectively. The percentage of MGE in the ICSI group (40.8 %) was significantly lower than that in the IVF group (56.1 %, p < 0.01). The percentages of BL and full-BL in the ICSI group were significantly lower than those in the IVF group. The pregnancy rates were similar in both the groups. CONCLUSIONS: Checking fertilization earlier than the usual period contributed to an avoidance of lower fertilization. Moreover, the embryos derived from ICSI after EFC possessed a normal developmental potential.

Effect of Male Cigarette Smoking on In Vitro Fertilization (IVF) Outcomes and Embryo Morphokinetic Parameters.

This retrospective cohort study examines the association between male smoking status and embryo development in vitro. The study included non-smoking women aged under 40 years who underwent in vitro fertilization (IVF) at Yanaihara Women's Clinic from May 2019 to May 2022, and they were divided into two groups according to the husband's smoking status. The effect of male smoking status on IVF outcomes was compared retrospectively based on embryonic development using a time-lapse incubator. A total of 184 patients were included; 272 oocytes of 45 female non-smokers were cultured with the sperm of male smokers, and 816 oocytes of 139 female non-smokers were cultured with the sperm of male non-smokers. No significant differences were observed between male smokers and non-smokers groups with regard to fertilization and the top-quality embryo on day 3 and day 5 (p > 0.05). The male smoker group's embryos reached the early cleavage-stage parameters (time of pronuclei appearance to the five-cell stage) significantly earlier than the male non-smoker group's embryos (p < 0.05). However, no significant differences were observed between the two groups in other parameters of top-quality blastocysts (p > 0.05). It was concluded that male smoking has some differences on the timing of early embryonic events on time-lapse examination.

Efficacy of Hyaluronan-Enriched Transfer Medium for Low-Grade Frozen Embryo Transfer.

RESEARCH QUESTION: This study aimed to retrospectively evaluate the efficacy of a hyaluronan-enriched transfer medium (HETM) for transfer failures and transfer of frozen embryos that have been graded as C at the time of transfer according to the Gardner classification of trophectoderm (TE). DESIGN: This study included 365 cycles of unsuccessful frozen-thawed embryo transfers in hormone replacement cycles graded C according to the Gardner classification of TE at the time of transfer. Clinical pregnancy rates were compared using the χ2 test, with the patients divided into two groups: one whose transfers did include HETM (HETM group) and one whose transfers did not include HETM (control group). As a subgroup analysis, patients with a TE grade of C at the time of transplantation were divided into two groups: those aged 39 years or younger and those aged 40 years or older at the time of transplantation. The clinical pregnancy rates of the groups with and without HETM were then compared. RESULTS: No difference in the clinical pregnancy rates between the HETM and control groups was observed. CONCLUSIONS: Hyaluronic acid is believed to favor implantation by promoting adhesion between the embryo and the endometrium, and there are reports of improved implantation and pregnancy rates as a result of HETM. However, the present results suggest limited effectiveness for HETM. Further case series should be conducted, and the suitability of its use as a treatment should be investigated.

Fertilization with human sperm bound to zona pellucida by pressing onto the oocyte membrane.

This study aimed to determine whether fertilization can be obtained by assisted fusion of oocyte and sperm without breaking the oocyte membrane. A total of 79 infertile couples, each with at least one unfertilized oocyte after in vitro fertilization (IVF), were recruited. Sperm collected from the zona pellucida (ZP) were pressed onto the membrane of unfertilized oocytes at either 6 h or 24 h after IVF, a procedure that we designated as assisted sperm fusion insemination (ASFI). The results of ASFI were compared with those obtained in a previous trial on oocytes in which rescue intracytoplasmic sperm injection (ICSI) was performed at 6 h after IVF. Acrosome reaction (AR) rate of sperm bound to ZP, fertilization rate, degeneration rate, and blastocyst formation rate were evaluated. The AR rate of sperm collected from the ZP was significantly higher than that of the motile sperm recovered from around the oocytes but not bound to the ZP after IVF (98.0% vs. 28.6%). ASFI which was performed at 6 h after IVF yielded a mean fertilization rate of 73.4% (58/79), a degeneration rate of 0% (0/79) and a blastocyst formation rate of 60.8% (31/51). Rescue ICSI which was performed at 6 h after IVF yielded a mean fertilization rate of 70.0% (70/100), a degeneration rate of 4% (4/100) and a blastocyst formation rate of 42.4% (25/59). Binding of sperm to the ZP typically results in AR. ASFI with acrosome-reacted sperm collected from the ZP yielded the fertilization rates similar to those obtained with rescue ICSI.

Mild stimulation with clomiphene citrate in combination with recombinant follicle-stimulating hormone and gonadotropin-releasing hormone antagonist and its influence on serum estradiol level and pregnancy rate.

Aim: The mild ovarian stimulation protocol for in vitro fertilization (IVF) is carried out to minimize adverse side-effects as well as cost. While performing mild ovarian stimulation with a gonadotropin-releasing hormone (GnRH) antagonist, the pregnancy rate was examined in cases that exhibited a serum estradiol (E2) drop down. Methods: In this study, 174 patients who requested mild ovarian stimulation for IVF began clomiphene citrate on day 3 and recombinant follicle-stimulating hormone (FSH) on day 5 of their menstrual cycles. A GnRH antagonist was administered when the dominant follicle reached a diameter of 14 mm. Serum luteinizing hormone and estradiol were measured at the time of GnRH antagonist administration and at the time of human chorionic gonadotropin (hCG) injection. Pregnancy rates and implantation rates were compared between 24 cycles in which the E2 level fell at the time of hCG injection and 150 cycles in which it did not fall. Results: The pregnancy rate in the cases in which the E2 level fell (25% decrease) at the time of hCG injection was significantly lower than it was in the cases in which it did not fall (16.7 vs 41.0%). The implantation rate for the cases in which the E2 level fell was also lower than that of the control group (7.0 vs 31.0%). There was no significant difference in the number of good-quality embryos between the two groups. Conclusion: When performing the mild ovarian stimulation protocol, serum E2 should be followed. It is prudent to avoid embryo transfer in the same cycle in cases that exhibit E2 drop down. (Reprod Med Biol 2008; 7: 85-89).

Sperm retention site and its influence on pronucleus stage evaluation following intracytoplasmic sperm injection.

Aim: It has been suggested that the position of the sperm after intracytoplasmic sperm injection (ICSI) has an effect on the development and quality of the embryo. In this study, we retrospectively examined whether pronucleus stage evaluation used through clinical studies in recent years has relevance with regard to sperm location. Methods: From 2003 to 2005, 1285 oocytes from 459 patients (average age: 36 years) were retrospectively analyzed. The 459 patients underwent ICSI because of fertilization disorders and oligozoospermia. Follicle stimulation was via either Clomid or the long protocol. Human chorionic gonadotropin was administered to induce ovulation and oocyte retrieval was conducted 35 h later. After confirming the presence of a polar body, we immobilized the ovum at the 6 o'clock position, introduced the injection pipette at the 3 o'clock position and carried out ICSI. Results: When a sperm was located at a position that was opposite to the polar body, both classifications of Scott and Tesarik regarding embryo quality were distinctly low. Furthermore, a good embryo classification ensued when the sperm was located adjacent to the polar body. Conclusion: The zone in which the sperm was located did not always correlate with embryo quality; however, our study suggested that sperm location affects the synchronization of the nucleolus. When carrying out ICSI, it is important to take into consideration the insertion point of the sperm. (Reprod Med Biol 2007; 6: 171-174).

Clinical outcomes of two different endometrial preparation methods for cryopreserved-thawed embryo transfer in patients with a normal menstrual cycle.

Aim: To compare the clinical outcomes of cryopreserved-thawed embryo transfer among patients with a normal menstrual cycle who had natural or hormone-replacement cycles. Methods: From January 2004 to June 2006, cryopreserved embryos following conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) were thawed and transferred in a total of 720 natural cycles and 136 hormone-replacement cycles. Results: Cryopreserved-thawed embryo transfer in patients who had a natural or hormone-replacement cycle resulted in clinical pregnancy in 43.1% and 40.4%, respectively; a rate of miscarriage of 14.5% and 23.6%, respectively; and a rate of ongoing pregnancy and delivery of 36.5% and 30.9%, respectively. None of these differences were statistically significant. Conclusions: Patients with a normal menstrual cycle who have natural or hormone-replacement cycles can be expected to have comparable clinical outcomes with cryopreserved-thawed embryo transfer. (Reprod Med Biol 2007; 6: 53-57).

Tablet terminals: a useful tool to explain in vitro fertilization treatment.

BACKGROUND: Explanations that involve medical care treatment take time. This also applies to explanations of in vitro fertilization (IVF) in the field of infertility treatment. This is because the cause of infertility differs from couple to couple, and because the explanations must begin with the mechanism of pregnancy. Recently, explanations facilitated by tablet terminals have been used in the field of medicine. In the present study, the efficacy and problems of explanations facilitated by tablet terminals were evaluated and compared with the explanations of IVF facilitated by paper-based pamphlets. METHODS: A total of 249 couples were asked to read a paper-based pamphlet explaining IVF treatment, while 252 couples were asked to view an explanation on a tablet terminal. The patients then answered a seven-item questionnaire. The answers to the questionnaire were based on a three-point scale, and statistical analysis was performed with the Mann-Whitney U test. RESULTS: Patients responded that the explanation facilitated by the tablet terminal was significantly easier to understand for all seven questionnaire items (p <0.05). The answer 'I did not understand' was selected for the items related to 'The treatment fees' (4.8% of answers) and 'Things to take note of, such as consultation times' (6.7% of answers). CONCLUSION: While patients generally did not understand the mechanism of pregnancy, explanations of IVF treatment facilitated by a tablet terminal were found to be more effective than paper-based explanations, although there is room for improvement.

Concentration of lactoferrin and interleukin-6 in cervical mucus from patients being treated for infertility.

Background: The concentrations of the iron-binding protein lactoferrin (LF) and interleukin-6 (IL-6) were measured in the cervical mucus of patients being treated for infertility throughout the menstrual cycle. Methods: A total of 251 cervical mucus samples were obtained from the patients throughout the menstrual cycle. One hundred and fifty samples were from primary infertility patients with unexplained infertility and 101 samples were from secondary infertility patients as a control. The concentrations of LF and IL-6 were measured by enzyme immunoassays. The standard curve of LF concentrations ranged from 1.6 to 50 ng/mL. Results: The mean LF and IL-6 concentrations in the cervical mucus of primary infertility patients were higher than that of the control patients (P= 0.04, P= 0.032, respectively) The LF and IL-6 concentrations were highly correlated (P < 0.0001). Conclusion: Elevated levels of IL-6 and LF in the cervical mucus were obtained from primary infertility patients. We speculate that LF might also be one of the causes of infertility and might play an important role in reproductive processes in the cervix. (Reprod Med Biol 2006; 5: 105-109).

Serum steroids concentration might be used for monitoring the growth of follicles in friendly in vitro fertilization.

Background: Steroid levels have been used as the predictive parameters for oocyte maturation and embryo development. In the present study, estradiol and progesterone concentrations in the follicular fluid and serum were evaluated in conventional in vitro fertilization (IVF; follicle stimulating hormone [FSH] and/or human menopausal gonadotropin [hMG] after pituitary desensitization) and friendly IVF (no stimulation, clomiphene citrate, small dose of FSH or hMG without pituitary desensitization). The purpose of the present study was to evaluate the differences in steroid distribution between conventional and friendly IVF. Methods: Concentrations of estradiol, progesterone, FSH, and luteinizing hormone (LH) in serum and follicular fluid were determined in conventional and friendly IVF protocols by an enzyme-linked immunosorbent assay kit. Correlations between follicular fluid and serum steroid concentrations in these different protocols, and between pregnant cycles and steroid concentrations were evaluated. Results: Two hundred and thirty-four samples of follicular fluid from 74 IVF patients were analyzed. In conventional IVF, there was no relationship in steroid levels in between follicular fluid and serum steroids, whereas serum steroid concentrations correlated with the number of developing follicles. There was a relationship between the serum and follicular fluid estradiol levels (r = 0.467, P < 0.0001) as well as progesterone levels (r = 0.227, P = 0.0488) from friendly IVF patients. Conclusions: Serum steroid concentrations were mainly associated with the number of developing follicles. In the cases of friendly IVF, which had a small number of developing follicles, serum steroids might be used to monitor follicular fluid steroid concentrations. (Reprod Med Biol 2006; 5: 277-282).

Differences in gene expression in the proliferative human endometrium.

OBJECTIVE: To use microdissection and DNA microarray technology to demonstrate differences in gene expression between epithelial and stromal areas in the proliferative human endometrium. DESIGN: Pilot study. SETTING: University hospital. PATIENT(S): Patients with normal menstrual cycles and at least one previous intrauterine pregnancy. INTERVENTION(S): Uterine endometrial biopsy. MAIN OUTCOME MEASURE(S): Gene expression. RESULT(S): From a total of 1,200 genes, 14 were strongly expressed in epithelial areas and 12 were strongly expressed in stromal areas. Among the genes strongly expressed in the stroma, expressions of decorin and discoidin domain receptor were confirmed by real-time polymerase chain reaction. Decorin was localized in the stromal areas by immunohistochemical staining. To confirm the effects of estrogen on gene expression, stromal cells were cultured. When E(2) was added to the culture media, expression of decorin mRNA was increased. CONCLUSION(S): The data demonstrated in this study help to understand the physiology of human endometrium. Decorin was strongly expressed in the stromal areas and was regulated by estrogen, and therefore it may be involved in restoration of the endometrium.

Expression of vascular endothelial growth factor (VEGF)-D and its receptor, VEGF receptor 3, as a prognostic factor in endometrial carcinoma.

PURPOSE: To evaluate the prognostic value of vascular endothelial growth factor (VEGF)-D and VEGF receptor (VEGFR)-3 in endometrial carcinoma. EXPERIMENTAL DESIGN: We assessed the levels of immunoreactivity for VEGF-D and VEGFR-3 in 71 endometrial carcinomas, 14 complex atypical endometrial hyperplasias, and 16 normal endometria by immunohistochemistry. RESULTS: VEGF-D was stained in both tumor cells and adjacent stromal cells. VEGFR-3 was stained in both tumor cells and adjacent endothelial cells. Immunoreactivity for VEGF-D in tumor cells and adjacent stromal cells became significantly stronger as lesions progressed from normal endometrium to advanced carcinoma. Similarly, immunoreactivity for VEGFR-3 in tumor cells and adjacent endothelial cells was significantly greater as lesions progressed from normal endometrium to advanced carcinoma. A strong correlation was found between high levels of VEGF-D immunoreactivity in carcinoma cells and VEGFR-3 in both carcinoma cells and adjacent endothelial cells. Similarly, high levels of VEGF-D immunoreactivity in stromal cells were significantly correlated with those of VEGFR-3 in both carcinoma cells and endothelial cells. High levels of VEGF-D in carcinoma cells and stromal cells, as well as those of VEGFR-3 in carcinoma cells and endothelial cells, were significantly related to myometrial invasion and lymph node metastasis. A strong correlation was found between poor survival and high levels of VEGF-D in both carcinoma cells and stromal cells and between poor survival and high levels of VEGFR-3 in carcinoma cells. Moreover, the high levels of VEGF-D in stromal cells and VEGFR-3 in carcinoma cells were independent prognostic factors in endometrial carcinoma. CONCLUSIONS: The presence of VEGF-D and VEGFR-3 in endometrial carcinoma may predict myometrial invasion and lymph node metastasis and may prospectively identify patients who are at increased risk for poor outcome. In addition, VEGF-D and VEGFR-3 may be promising targets for new therapeutic strategies in endometrial carcinoma.

Comparison in gene expression of secretory human endometrium using laser microdissection.

BACKGROUND: The endometrium prepares for implantation under the control of steroid hormones. It has been suggested that there are complicated interactions between the epithelium and stroma in the endometrium during menstrual cycle. In this study, we demonstrate a difference in gene expression between the epithelial and stromal areas of the secretory human endometrium using microdissection and macroarray technique. METHODS: The epithelial and stromal areas were microdissected from the human endometrium during the secretory phase. RNA was extracted and amplified by PCR. Macroarray analysis of nearly 1000 human genes was carried out in this study. Some genes identified by macroarray analysis were verified using real-time PCR. RESULTS: In this study, changes in expression <2.5-fold in three samples were excluded. A total of 28 genes displayed changes in expression from array data. Fifteen genes were strongly expressed in the epithelial areas, while 13 genes were strongly expressed in the stromal areas. The strongly expressed genes in the epithelial areas with a changes >5-fold were WAP four-disulfide core domain 2 (44.1 fold), matrix metalloproteinase 7 (40.1 fold), homeo box B5 (19.8 fold), msh homeo box homolog (18.8 fold), homeo box B7 (12.7 fold) and protein kinase C, theta (6.4 fold). On the other hand, decorin (55.6 fold), discoidin domain receptor member 2 (17.3 fold), tissue inhibitor of metalloproteinase 1 (9 fold), ribosomal protein S3A (6.3 fold), and tyrosine kinase with immunoglobulin and epidermal growth factor homology domains (5.2 fold) were strongly expressed in the stromal areas. WAP four-disulfide core domain 2 (19.4 fold), matrix metalloproteinase 7 (9.7-fold), decorin (16.3-fold) and tissue inhibitor of metalloproteinase 1 (7.2-fold) were verified by real-time PCR. CONCLUSIONS: Some of the genes we identified with differential expression are related to the immune system. These results are telling us the new information for understanding the secretory human endometrium.

Regulation of estrogen activity in human endometrium: effect of IL-1beta on steroid sulfatase activity in human endometrial stromal cells.

We investigated the effect of interleukin 1beta (IL-1beta) on steroid sulfatase (STS) activity and the expression of STS mRNA in human endometrial stromal cells. Endometrial tissue samples were obtained from patients undergoing hysterectomy to remove uterine fibroids. Stromal cells were isolated from the tissue preparation and cultured. IL-lbeta (1 approximately 100 ng/ml) was added into the culture medium and incubated for 24 h. The expression of STS mRNA was measured by competitive RT-PCR. The addition of IL-lbeta at 10 and 100 ng/ml suppressed STS mRNA expression to 55.2 +/- 12.8% and 25.1 +/- 10.9%, respectively, of the control sample to which no IL-lbeta had been added. STS activity was measured by radiolabelled steroid metabolite using thin layer chromatography, and this activity was also significantly suppressed in response to the administration of IL-lbeta in a dose-dependent manner. When IL-1 receptor antagonist (IL-1ra) was added together with IL-1beta to the culture medium, mRNA expression and STS activity were recovered. The present study is the first to demonstrate IL-1beta regulation of STS activity locally in human endometrium. IL-1beta suppressed mRNA and activity of STS in stromal cell culture. This initial demonstration of IL-1beta regulation of STS implies that IL-1beta may control the steroid microenvironment in human uterine endometrium by reducing biologic action of estrogen.

Correlations between steroids concentration in follicular fluid, pronuclear morphology and embryo qualities in in vitro fertilization.

Background: Several parameters of early embryo development are known as predictors of implantation success. Recently, zygote or embryo morphological assessments are thought to be a major method of selection in embryo transfer. We expected that the concentrations of the steroids in follicular fluid (FF) were associated with oocyte maturation and embryo quality. In the present paper, we evaluated the relationship of several parameters. Methods: We investigated 105 samples of FF from 22 subjects by in vitro fertilization (IVF). We evaluated the correlations between the FF concentrations of estradiol (E2) and progesterone (P4), the diameter of the ovarian follicles, fertilization, and zygote assessment based on pronuclear morphology and day 3 embryo qualities (i.e. number of blastomeres and fragmentation rate). Results: There was a positive correlation between the E2 concentrations in FF and serum (r = 0.273, P < 0.01), but there was no correlation between follicular diameter and the FF concentration of each steroid. The concentration of E2 in FF containing fertilized oocytes was not significantly different from that in FF containing unfertilized oocytes. At the pronuclear stage, the concentration of either steroid in FF did not differ among the morphological groups. The concentration of P4 in FF was significantly lower in the group in which pronuclei were detected at 20 h after insemination than in the group in which pronuclei were not detected. The concentration of E2 in FF was significantly related to the number of blastomeres (r = 0.271, P < 0.05) and furthermore, was significantly higher in FF from which morphologically good embryos were obtained at day 3 (P < 0.05). Conclusions: The FF concentrations of the steroids did not affect the pronuclear pattern, but P4 production may play a role in reducing the potential of the oocyte to develop pronuclei and the concentration of E2 may predict the cleavage capability of the oocyte. (Reprod Med Biol 2003; 2: 171-176).