Comparison of clinical and preimplantation genetic testing for aneuploidy outcomes between in vitro fertilization and intracytoplasmic sperm injection in sibling mature oocytes from high-risk patients: A retrospective study.

AIM: To evaluate the influence of insemination methods on clinical outcomes by assessing preimplantation genetic testing for aneuploidy (PGT-A) outcomes in embryos obtained using in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in sibling mature oocytes from high-risk patients. METHODS: This retrospective study involved 108 couples with nonmale or mild male factor infertility who underwent split insemination cycles from January 2018 to December 2021. PGT-A was performed using trophectoderm biopsy, array comparative genome hybridization, or next-generation sequencing with 24-chromosome screening. RESULTS: Mature oocytes were divided into IVF (n = 660) and ICSI (n = 1028) groups. The normal fertilization incidence was similar between the groups (81.1% vs. 84.6%). The total number of blastocysts biopsied was significantly higher in the IVF group than in the ICSI group (59.3% vs. 52.6%; p = 0.018). However, euploidy (34.4% vs. 31.9%) and aneuploidy (63.4% vs. 66.2%) rates per biopsy and clinical pregnancy rates (60.0% vs. 58.8%) were similar between the groups. Implantation (45.6% vs. 50.8%) and live birth or ongoing pregnancy (52.0% vs 58.8%) rates were slightly higher in the ICSI group than in the IVF group and miscarriage rate per transfer was slightly higher in the IVF group than in the ICSI group (12.0% vs 5.9%); however no significant difference was observed. CONCLUSIONS: IVF and ICSI using sibling mature oocytes had similar clinical outcomes, and euploidy and aneuploidy rates in couples with nonmale and mild male factor infertility. These results suggest that IVF is a useful option, along with ICSI, as an insemination method in PGT-A cycles, especially in high-risk patients.

IVM of human immature oocytes for infertility treatment and fertility preservation.

Background: Thousands of healthy babies are born from in vitro maturation (IVM) procedures, but the rate of efficiency differs with the source of immature oocytes obtained. Recently, there are different IVM protocols proposed for infertility treatment and fertility preservation. Methods: Based on the literature, the clinical application for IVM of immature oocytes was summarized. Main findings Results: Immature oocytes may be retrieved from women after priming with or without the use of follicular stimulation hormone (FSH), human chorionic gonadotrophin (hCG) or a combination of both FSH and hCG. Successful pregnancy rates with IVM technology seem to be correlated with the number of immature oocytes obtained. With the source and culture course of immature oocytes, there are various IVM protocols. IVM of immature oocytes is profoundly affected by the culture conditions, but no breakthrough has been made by improving the IVM medium itself. Thus, the clinical application of IVM technology continues to evolve. Conclusion: IVM technology is a useful technique for infertile women and fertility preservation. Mild stimulation IVF combined with IVM of immature oocytes is a viable alternative to the conventional stimulation IVF cycle treatment as it may prove to be an optimal first-line treatment approach.

RANKL down-regulates the mast cell proliferation through inducing senescence.

An increase in the number of mast cells could contribute to inflammatory diseases and pathologic conditions. A receptor activator of NF-κB ligand (RANKL)/RANK system is one of the key signaling pathways accelerating mast cell-mediated allergic inflammatory reactions. However, the biological functions of RANKL in mast cell proliferation remains to be clarified. The aim of the present study is to clarify the role of RANKL in mast cell proliferation. Surprisingly, RANKL remarkably reduced the proliferation of human mast cell line, HMC-1 cells through the inhibition of murine double minute 2 (MDM2) and Ki-67 mRNA expressions in a dose-dependent manner. RANKL significantly reduced cell viability, whereas it increased cellular senescence via increasing levels of p53, phosphorylated(p)-p53, p21, and p16 and decreasing levels of retinoblastoma protein (pRb) and p-pRb in HMC-1 cells. Even in rat peritoneal mast cells, RANKL induced cellular senescence by increasing filamentous-actin polymerization. In addition, RANKL remarkably reduced thymic stromal lymphopoietin (TSLP)-induced mast cell proliferation via the downregulation of MDM2 and Ki-67. RANKL decreased levels of p-signal transducer and activator of transcription 6 in TSLP-stimulated HMC-1 cells. The mast cell growth factor, interleukin-13 was remarkably down-regulated by treatment with RANKL in TSLP-stimulated HMC-1 cells. Furthermore, RANKL increased the number of senescence-associated ß-galactosidase-stained cells and protein levels of p53, p-p53, and p21 in TSLP-stimulated HMC-1 cells. These data suggest that RANKL down-regulates mast cell proliferation by inducing senescence.

A calcium channel blocker, manoalide exerts an anti-allergic inflammatory effect through attenuating NF-κB activity.

BACKGROUND: Many people are troubled by allergic inflammation including ocular allergic diseases, anaphylaxis, allergic rhinitis, atopic dermatitis, and eczema. Consequently, finding medications for use in allergic inflammation therapy is crucial in human health. Manoalide, a marine natural product isolated as an anti-bacterial metabolite from Luffariella variabilis, is a calcium channel blocker. However, its latent ability as an anti-allergic inflammatory agent has not yet been reported. Our research aimed to elucidate whether manoalide exerts an anti-allergic inflammatory effect in the human mast cell line, HMC-1. METHODS: Herein, we investigated the immunoregulatory effects and molecular mechanisms of manoalide in HMC-1 cells. RESULTS: Manoalide significantly alleviated secretion of the inflammatory cytokines interleukin (IL)-1ß, thymic stromal lymphopoietin, tumor necrosis factor-α, IL-6, and IL-8 via blockage of caspase-1 without cytotoxicity in activated HMC-1 cells. Activation of nuclear factor-κB increased by mast cell stimulation was attenuated by treatment with manoalide. In addition, we demonstrated that manoalide treatment remarkably attenuated the activation of mitogen-activated protein kinases in activated-HMC-1 cells. CONCLUSIONS: Taken together, our findings indicate manoalide has an anti-allergic inflammatory role, and we propose that manoalide might have potential as a novel anti-allergic inflammatory agent.

The effects of luteinising hormone gene polymorphism on the outcomes of in vitro fertilisation and embryo transfer.

Trp8Arg polymorphism of the LH beta gene has decreased bioactivity in vivo and previous studies showed conflicting data on the effect of LH beta gene polymorphism on the IVF outcome. In this study, 591 IVF patients were recruited. Patients with the variant allele(s) were the carrier group. In GnRH antagonist cycles, the clinical pregnancy rate was significantly lower in the carrier group (18.9%) than in the noncarrier group (37.1%). In long GnRH agonist cycles, the clinical pregnancy rate was comparable between both groups. To clarify the effect of COH protocols, IVF outcomes in the GnRH antagonist and long GnRH agonist protocol groups in carriers were analysed. Among carriers, the clinical pregnancy rate was significantly lower in the GnRH antagonist protocol group (18.9%) than in the long GnRH agonist protocol group (45.2%). Single nucleotide polymorphism analysis may contribute to the individualisation of COH protocols for each patient in the future.Impact StatementWhat is already known on this subject? Trp8Arg polymorphism of the LH beta gene is known to have decreased bioactivity in vivo. Previous studies have demonstrated hypo-sensitivity in the patients with the variant LH beta protein, while other study showed similar carrier frequency between the poor and the normal response group.What the results of this study add? The variant LH beta gene was associated with a lower clinical pregnancy rate in GnRH antagonist cycles but not in long GnRH agonist cycles.What the implications are of these findings for clinical practice and/or further research? Single nucleotide polymorphism analysis may contribute to the individualisation of COH protocols for each patient in the future.

A Comparison of Embryonic Development and Clinical Outcomes between In vitro Oocytes Maturation Using Micro-Vibration System and In vivo Oocytes Maturation in Polycystic Ovarian Syndrome Patients.

BACKGROUND/OBJECTIVES: Mechanical micro-vibration remains insufficient for improving embryo culture conditions in human immature oocytes. This study compared the clinical outcomes and embryo development between germinal vesicle (GV) oocytes with the micro-vibration culture (MVC) system in in vitro maturation (IVM) cycles and in vivo-matured oocytes in controlled ovarian hyperstimulation (COH) cycles in polycystic ovarian syndrome (PCOS) patients. METHODS: This study investigated 152 PCOS patients who underwent 159 fresh embryo transfer cycles, including IVM cycles with embryos derived from GV oocytes and the COH cycles with embryos derived from in vivo-matured oocytes. The IVM cycles were divided into groups according to the culture system used: static culture (SC) and MVC: In the IVM-S group (n = 47), SC was applied during both IVM and in vitro culture (IVC), whereas in the IVM-MV group (n = 44), MVC was applied during both IVM and IVC. For the COH cycles, in the COH-S group (n = 68), SC was applied during IVC. RESULTS: The number of in vitro-matured oocytes was similar in the IVM-S and IVM-MV groups, but the good-quality embryo (GQE; ≥6-cells) rate was significantly higher in the IVM-MV group (p < 0.01). The GQE rate and clinical outcomes of the COH-S group were significantly better than those of the IVM-S group (p < 0.05) but similar to those of the IVM-MV group. CONCLUSION: Compared with the SC system, the MVC system in IVM cycles improves the embryonic quality of GV oocytes and clinical outcomes, resulting in development of potential equivalent to in vivo-matured oocytes.

Sulforaphane mitigates mast cell-mediated allergic inflammatory reactions in in silico simulation and in vitro models.

Objectives: Sulforaphane, a major ingredient isolated from Brassica oleracea var. italica (broccoli), is known to exhibit anti-inflammatory, anti-cancer, and anti-diabetic effects. In this study, we employed an in vitro model of phorbol 12-myristate 13-acetate and a23187 (PMACI)-stimulated human mast cells (HMC-1 cells) to investigate the anti-allergic inflammatory effects and mechanisms of sulforaphane and Brassica oleracea var. italica extracts.Methods: Cytokine levels were measured by ELISA and quantitative real-time-PCR methods. Caspase-1 activity was determined by caspase-1 assay. Binding mode of sulforaphane within caspase-1 was determined by molecular docking simulation. Protein expression was determined by Western blotting.Results: Water extract of Brassica oleracea var. italica (WE) significantly reduced thymic stromal lymphopoietin (TSLP) secretion and caspase-1 activity on activated HMC-1 cells. In the molecular docking simulation and in vitro caspase-1 assays, sulforaphane regulated caspase-1 activity by docking with the identical binding site of caspase-1. Sulforaphane significantly inhibited the levels of inflammatory mediators including TSLP, tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-8 in a dose-dependent manner. Immunoblotting experiments revealed that sulforaphane and WE reduced translocation of NF-κBp65 into the nucleus and phosphorylation of IκBα in the cytosol. Furthermore, phosphorylation of mitogen-activated protein kinases (MAPK) was down-regulated by treatment with sulforaphane or WE.Conclusion: Our findings suggest that sulforaphane and WE have anti-allergic inflammatory effects by intercepting caspase-1/NF-κB/MAPKs signaling pathways.

Successful pregnancy and delivery after ICSI with artificial oocyte activation by calcium ionophore in in-vitro matured oocytes: a case report.

The achievement of a successful pregnancy and delivery after oocyte activation with calcium ionophore is reported in a couple having low fertilization rates after intracytoplasmic sperm injection (ICSI) of in-vitro matured oocytes. A couple, in which the wife had polycystic ovary syndrome and the husband had moderate oligoteratozoospermia, showed a low fertilization rate in a previous in-vitro maturation cycle (2/11 [18.2%]). The most likely cause of complete fertilization failure or low fertilization rates is failure of oocyte activation. Therefore, artificial oocyte activation by calcium ionophore was combined with ICSI to achieve viable fertilized oocytes. Oocytes were stimulated with calcium ionophore for 30 min after ICSI. The fertilization rate of oocytes activated with calcium ionophore (13/15 [86.7%] and 7/9 [77.8%]) was higher than that of the non-activated oocytes. In the latest cycle, three embryos derived from the activated oocytes were transferred into the uterus on day 3. Subsequently, two gestational sacs were identified on ultrasound. The patient delivered dizygotic twins (girl 2260 g and boy 2760 g) at 35 weeks and 6 days gestation by caesarean section. This result suggests that calcium ionophore could be useful for oocyte fertilization in couples with low fertilization rates after ICSI of in-vitro matured oocytes.

Development of a security system for assisted reproductive technology (ART).

PURPOSE: In the field of assisted reproductive technology (ART), medical accidents can result in serious legal and social consequences. This study was conducted to develop a security system (called IVF-guardian; IG) that could prevent mismatching or mix-ups in ART. MATERIALS AND METHODS: A software program was developed in collaboration with outside computer programmers. A quick response (QR) code was used to identify the patients, gametes and embryos in a format that was printed on a label. There was a possibility that embryo development could be affected by volatile organic components (VOC) in the printing material and adhesive material in the label paper. Further, LED light was used as the light source to recognize the QR code. Using mouse embryos, the effects of the label paper and LED light were examined. The stability of IG was assessed when applied in clinical practice after developing the system. A total of 104 cycles formed the study group, and 82 cycles (from patients who did not want to use IG because of safety concerns and lack of confidence in the security system) to which IG was not applied comprised the control group. RESULTS: Many of the label paper samples were toxic to mouse embryo development. We selected a particular label paper (P touch label) that did not affect mouse embryo development. The LED lights were non-toxic to the development of the mouse embryos under any experimental conditions. There were no differences in the clinical pregnancy rates between the IG-applied group and the control group (40/104 = 38.5 % and 30/82 = 36.6 %, respectively). CONCLUSIONS: The application of IG in clinical practice did not affect human embryo development or clinical outcomes. The use of IG reduces the misspelling of patient names. Using IG, there was a disadvantage in that each treatment step became more complicated, but the medical staff improved and became sufficiently confident in ART to offset this disadvantage. Patients who received treatment using the IG system also went through a somewhat tedious process, but there were no complaints. These patients gained further confidence in the practitioners over the course of treatment.

Clinical outcomes of preimplantation genetic testing for aneuploidy in high-risk patients: a retrospective cohort study.

OBJECTIVE: The purpose of this study was to evaluate the impact of preimplantation genetic testing for aneuploidy (PGT-A) on clinical outcomes among high-risk patients. METHODS: This retrospective study involved 1,368 patients and the same number of cycles, including 520 cycles with PGT-A and 848 cycles without PGT-A. The study participants comprised women of advanced maternal age (AMA) and those affected by recurrent implantation failure (RIF), recurrent pregnancy loss (RPL), or severe male factor infertility (SMF). RESULTS: PGT-A was associated with significant improvements in the implantation rate (IR) and the ongoing pregnancy rate/live birth rate (OPR/LBR) per embryo transfer cycle in the AMA (39.3% vs. 16.2% [p<0.001] and 42.0% vs. 21.8% [p<0.001], respectively), RIF (41.7% vs. 22.0% [p<0.001] and 47.0% vs. 28.6% [p<0.001], respectively), and RPL (45.6% vs. 19.5% [p<0.001] and 49.1% vs. 24.2% [p<0.001], respectively) groups, as well as the IR in the SMF group (43.3% vs. 26.5%, p=0.011). Additionally, PGT-A was associated with lower overall incidence rates of early pregnancy loss in the AMA (16.7% vs. 34.3%, p=0.001) and RPL (16.7% vs. 50.0%, p<0.001) groups. However, the OPR/LBR per total cycle across all PGT-A groups did not significantly exceed that for the non-PGT-A groups. CONCLUSION: PGT-A demonstrated beneficial effects in high-risk patients. However, our findings indicate that these benefits are more pronounced in carefully selected candidates than in the entire high-risk patient population.

Effects of oxygen tension and IGF-I on HIF-1α protein expression in mouse blastocysts.

PURPOSE: Hypoxia inducible factors (HIFs) are key regulators of oxygen homeostasis in response to reduced oxygenation in somatic cells. In addition, HIF-1α protein can be also induced by insulin-like growth factor I (IGF-I) treatment in various cell lines under normoxic condition. However, the expression and function of HIF-1α in embryogenesis are still unclear. Therefore, the objectives of this study were to examine the expression of HIF-1α in mouse blastocysts cultured under hypoxic and normoxic conditions, and to determine whether oxygen tension and IGF-I influence embryonic development through stimulation of HIF-1α expression. METHODS: Mouse embryos were cultured from the 1-cell to blastocyst stage under 5 % or 20 % O(2) in both the absence and presence of IGF-I. RESULTS: The embryonic development rates to the blastocyst stage were not affected by oxygen tension or IGF-I treatment. HIF-1α protein was localized to the cytoplasm of blastocysts, and its levels were independent of oxygen concentration or IGF-I treatment. Blastocysts cultured under 5 % O(2) exhibited significantly higher total cell numbers (83.4 ± 18.1) and lower apoptotic index (3.7 ± 1.5) than those cultured under 20 % O(2) (67.4 ± 15.6) (6.9 ± 3.5) (P<0.05). IGF-I reduced the apoptotic index in both oxygen conditions, but a significant decrease was detected in the 20 % O(2) group. CONCLUSIONS: HIF-1α may not be a major mediator that responds to change in oxygen tension within blastocysts, inconsistent with that of somatic cells. Supplementation of culture media with IGF-I has been shown to promote embryo development by an anti-apoptotic effect, instead of increasing HIF-1α protein expression.

Comparison of clinical outcomes between single and double vitrified-warmed blastocyst embryo transfer according to the day of vitrification.

PURPOSE: To compare the efficacy of single vitrified-warmed blastocyst embryo transfer (SVBT) versus double vitrified-warmed blastocyst embryo transfer (DVBT) according to the day of vitrification. METHODS: This retrospective study included a total of 1,051 cycles in women less than 37 years of age with their autologous SVBT cryopreserved on day 5 (5d-SVBT, n = 737) or day 6 (6d-SVBT, n = 154) and DVBT on day 5 (5d-DVBT, n = 129) or day 6 (6d-DVBT, n = 31) from January 2009 to December 2011. RESULTS: The clinical pregnancy rate (41.8 % vs. 48.1 %, p = 0.184) and ongoing pregnancy rate (36.6 % vs. 45.0 %, p = 0.072) were not significantly different between the 5d-SVBT group and the 5d-DVBT group. However, the clinical pregnancy (29.9 % vs. 58.1 %, p = 0.003) and ongoing pregnancy rates (23.4 % vs. 51.6 %, p = 0.001) were significantly lower in the 6d-SVBT group compared with those in the 6d-DVBT group. The implantation rate (42.2 % vs. 34.5 %, p = 0.03) of the 5d-SVBT group was significantly higher than that of the 5d-DVBT group, while the implantation rate (29.9 % vs. 37.1 %, p = 0.303) of the 6d-SVBT group was not statistically different compared with that in the 6d-DVBT group. The multiple pregnancy rates (1.0 % in the 5d-SVBT group vs. 38.7 % in the 5d-DVBT group, p < 0.001 and 0 % in the 6d-SVBT group vs. 22.2 % in the 6d-DVBT group, p = 0.001) were statistically significantly lower in the SVBT group compared with those in the DVBT group regardless of the day of vitrification. CONCLUSIONS: This study showed that the 5d-SVBT resulted in comparable clinical outcomes compared to the 5d-DVBT while the 6d-SVBT yielded significantly lower clinical outcomes compared to the 6d-DVBT.

Effect of micro-vibration culture system on embryo development.

PURPOSE: Micro-vibration culture system was examined to determine the effects on mouse and human embryo development and possible improvement of clinical outcomes in poor responders. MATERIALS AND METHODS: The embryonic development rates and cell numbers of blastocysts were compared between a static culture group (n = 178) and a micro-vibration culture group (n = 181) in mice. The embryonic development rates and clinical results were compared between a static culture group (n = 159 cycles) and a micro-vibration culture group (n = 166 cycles) in poor responders. A micro-vibrator was set at a frequency of 42 Hz, 5 s/60 min duration for mouse and human embryo development. RESULTS: The embryonic development rate was significantly improved in the micro-vibration culture group in mice (p < 0.05). The cell numbers of mouse blastocysts were significantly higher in the micro-vibration group than in the static culture group (p < 0.05). In the poor responders, the rate of high grade embryos was not significantly improved in the micro-vibration culture group on day 3. However, the optimal embryonic development rate on day 5 was improved in the micro-vibration group, and the total pregnancy rate and implantation rate were significantly higher in the micro-vibration group than in the static culture group (p < 0.05). CONCLUSIONS: Micro-vibration culture methods have a beneficial effect on embryonic development in mouse embryos. In poor responders, the embryo development rate was improved to a limited extent under the micro-vibration culture conditions, but the clinical results were significantly improved.

Analysis of clinical outcomes with respect to spermatozoan origin after artificial oocyte activation with a calcium ionophore.

PURPOSE: Fertilization failures have occurred repeatedly in reproductive centers after intracytoplasmic sperm injection (ICSI) and artificial oocyte activation (AOA) has been used to prevent it. This study was performed to investigate whether spermatozoan origin influences clinical outcomes of AOA with a calcium ionophore. METHODS: A total of 185 ICSI cycles with a history of no or low fertilization was included in this retrospective study. The outcomes of AOA after ICSI were compared with ejaculated-normal, ejaculated-oligo-astheno-terato or extracted-testicular spermatozoa. RESULTS: There were significant differences between the previous standard ICSI cycles and AOA cycles in the rate of fertilization and clinical outcomes among cases with different sperm origins. Thirty-eight healthy babies (20 singles and 18 twins, 29 cycles) were successfully delivered, and no congenital birth defects were observed. CONCLUSIONS: Most patients with a no or low fertilization history obtained an increased fertilization rate and a positive clinical outcome with AOA regardless of the origin of spermatozoa.

Anti-fatigue effect of tormentic acid through alleviating oxidative stress and energy metabolism-modulating property in C2C12 cells and animal models.

BACKGROUND/OBJECTIVES: Oxidative stress is caused by reactive oxygen species and free radicals that accelerate inflammatory responses and exacerbate fatigue. Tormentic acid (TA) has antioxidant and anti-inflammatory properties. Thus, the aim of present study is to determine the fatigue-regulatory effects of TA in H2O2-stimulated myoblast cell line, C2C12 cells and treadmill stress test (TST) and forced swimming test (FST) animal models. MATERIALS/METHODS: In the in vitro study, C2C12 cells were pretreated with TA before stimulation with H2O2. Then, malondialdehyde (MDA), lactate dehydrogenase (LDH), creatine kinase (CK) activity, tumor necrosis factor (TNF)-α, interleukin (IL)-6, superoxide dismutase (SOD), catalase (CAT), glycogen, and cell viability were analyzed. In the in vivo study, the ICR male mice were administered TA or distilled water orally daily for 28 days. FST and TST were then performed on the last day. In addition, biochemical analysis of the serum, muscle, and liver was performed. RESULTS: TA dose-dependently alleviated the levels of MDA, LDH, CK activity, TNF-α, and IL-6 in H2O2-stimulated C2C12 cells without affecting the cytotoxicity. TA increased the SOD and CAT activities and the glycogen levels in H2O2-stimulated C2C12 cells. In TST and FST animal models, TA decreased the FST immobility time significantly while increasing the TST exhaustion time without weight fluctuations. The in vivo studies showed that the levels of SOD, CAT, citrate synthase, glycogen, and free fatty acid were increased by TA administration, whereas TA significantly reduced the levels of glucose, MDA, LDH, lactate, CK, inflammatory cytokines, alanine transaminase, aspartate transaminase, blood urea nitrogen, and cortisol compared to the control group. CONCLUSIONS: TA improves fatigue by modulating oxidative stress and energy metabolism in C2C12 cells and animal models. Therefore, we suggest that TA can be a powerful substance in healthy functional foods and therapeutics to improve fatigue.

Live birth after SrCl(2) oocyte activation in previous repeated failed or low fertilization rates after ICSI of frozen-thawed testicular spermatozoa: case report.

PURPOSE: To report a live birth resulting after strontium chloride (SrCl(2)) oocyte activation in a couple with complete fertilization failure or low fertilization rates following intracytoplasmic sperm injection (ICSI) of frozen-thawed testicular spermatozoa. METHODS: The couple underwent ICSI of frozen-thawed testicular spermatozoa. After ICSI, the oocytes were artificially activated by SrCl(2) because the results of fertilization were not satisfactory in the previous cycles. The main outcome measures were fertilization, pregnancy, and birth. RESULTS: In the first and second cycles performed previously at another clinic, fertilization rates were 9.1 % and 0.0 %, respectively. In the third cycle, 31 metaphase II oocytes were retrieved. After sperm injection, all of the oocytes were stimulated using SrCl(2) for activation. Sixteen oocytes were fertilized (51.6 %), and a single embryo was transferred into the uterus on Day 3. A healthy girl weighing 2750 g was born at 40 weeks of gestation by caesarean section. CONCLUSIONS: This result suggests that SrCl(2) could be useful for oocyte fertilization in case of repeated complete fertilization failure or low fertilization rates following ICSI of frozen-thawed testicular spermatozoa.

Clinical outcomes of elective single morula embryo transfer versus elective single blastocyst embryo transfer in IVF-ET.

PURPOSE: To compare the clinical outcomes of elective single morula embryo transfer (eSMET) versus elective single blastocyst embryo transfer (eSBET) in selected patients. METHODS: This study was a retrospective study which analyzed for 271 cycles in women under 37 years of age who are undergoing their first or second trial of in vitro fertilization-embryo transfer (IVF-ET) from January 2008 to December 2009. The eSMET was performed on day 4 (n = 130) and the eSBET was conducted on day 5 (n = 141). RESULTS: The clinical pregnancy rate (51.5% vs. 51.8%, p = 0.97), implantation rate (52.3% vs. 52.5%, p = 0.98), and live birth rate (39.2% vs. 44.7%, p = 0.36) were similar in the eSMET and eSBET groups, respectively. The miscarriage rate of the eSMET group (23.9%) was slightly higher than that of the eSBET group (13.7%) (p = 0.12), without reaching statistical significance. There was only one case of monozygotic twin pregnancy in each group. CONCLUSIONS: The clinical outcomes of day 4 eSMET were comparable to those of day 5 eSBET. Therefore, day 4 eSMET is a viable option or an alternative to day 5 eSBET, with no difference in success rates.

Dp44mT regulates the levels of inflammatory mediators through blocking NF-κB nuclear translocation in LPS-stimulated RAW 264.7 macrophages.

Inflammation is increased by infection with pathogens such as viruses, bacteria, and parasites. High levels of inflammatory mediators and infiltration of macrophages into inflammatory lesions were reported in severe inflammatory diseases. Here, the aim of this study was to evaluate an anti-inflammatory activity of di-2-pyridylketone-4,4-dimethyl-3-thiosemicarbazone (Dp44mT) on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Dp44mT (1-100 ng/mL) had no effect on viability of RAW 264.7 macrophages. Dp44mT (100 ng/mL) significantly reduced LPS-induced release of nitric oxide and expression of inducible nitric oxide synthase and cyclooxygenase-2. A significant upregulation of tumor necrosis factor (TNF)-α and interleukin (IL)-6 by LPS stimulation was downregulated by treatment with Dp44mT. Dp44mT blocked activation of nuclear factor-κB by the interruption of IκBα phosphorylation. Dp44mT suppressed the phagocytosis. Furthermore, administration of Dp44mT significantly reduced the serum levels of TNF-α and IL-6 in LPS-treated mice without side effects. In conclusion, these results indicate that Dp44mT has an anti-inflammatory activity and may be of therapeutic significant for the prevention and treatment of inflammatory diseases.

Healthy live birth from vitrified blastocysts produced from natural cycle IVF/IVM.

Natural-cycle IVF combined with in-vitro maturation (natural-cycle IVF/IVM) was used as a treatment for a 27-year-old woman. She was administered 10,000 IU of human chorionic gonadotrophin intramuscularly about 36 h prior to oocyte collection and oocyte collection was performed on day 11 of her menstrual cycle. One mature oocyte was retrieved from the leading follicle and another five mature oocytes and six immature oocytes were retrieved from the rest of the follicles. Out of 10 fertilized zygotes, eight of them cleaved. Three day-3 embryos derived from in-vivo matured oocytes (one was from the leading follicle) were transferred but failed to conceive. The remaining five embryos were continuously cultured until day 6 and four of them developed to the expanded blastocyst stage and vitrified for the storage. Six months later, two vitrified-warmed blastocysts derived from the immature oocytes were transferred and resulted in the full-term delivery of a healthy female infant. This case report for the first time indicates that blastocysts produced from the immature oocytes retrieved from the small follicles, when a leading follicle exists in the ovaries, can be vitrified to produce a healthy live birth, suggesting that natural-cycle IVF/IVM is an efficient infertility treatment.

Effects of resveratrol, granulocyte-macrophage colony-stimulating factor or dichloroacetic acid in the culture media on embryonic development and pregnancy rates in aged mice.

The success rate of assisted reproductive technology is closely correlated with maternal age. Reproductive aging pathologies are frequently caused by impaired DNA repair, genomic instability, and mitochondrial dysfunction. Several reports have shown that resveratrol can prevent age-related diseases by improving mitochondrial function. Improved blastocyst development and mitochondrial output by dichloroacetic acid (DCA) supplementation were reported in aged mice. Granulocyte-macrophage colony-stimulating factor (GM-CSF) has significant effects on implantation rates in women with previous miscarriages. Therefore, this study was conducted to observe how those compounds influence the developmental and the reproductive potential of aged oocytes. BDF1 female mice at 58-62 weeks old were used for this study. MII oocytes were fertilized and cultured in MRC media supplemented with or without resveratrol (0.5 µM), GM-CSF (2 ng/ml) or DCA (1.0 mM). The addition of resveratrol, GM-CSF or DCA tended to increase blastocyst development and pregnancy rates. Supplementation with resveratrol significantly increased the pregnancy and implantation rates (p < 0.05). Moreover, resveratrol decreased reactive oxygen species production and increased mitochondrial membrane potential. These results suggest that the addition of resveratrol can increase pregnancy outcomes in women of advanced maternal age.