The Presence of TGFß3 in Human Ovarian Intrafollicular Fluid and Its Involvement in Thromboxane Generation in Follicular Granulosa Cells through a Canonical TGFßRI, Smad2/3 Signaling Pathway and COX-2 Induction.

Ovarian follicular fluid (FF) has a direct impact on oocyte quality, playing key roles in fertilization, implantation, and early embryo development. In our recent study, we found FF thromboxane (TX) to be a novel factor inversely correlated with oocyte maturation and identified thrombin, transforming growth factor ß (TGFß), TNF-α, and follicular granulosa cells (GCs) as possible contributors to FF TX production. Therefore, this study sought to investigate the role of TGFß3 in regulating TX generation in human ovarian follicular GCs. TGFß3 was differentially and significantly present in the FF of large and small follicles obtained from IVF patients with average concentrations of 68.58 ± 12.38 and 112.55 ± 14.82 pg/mL, respectively, and its levels were correlated with oocyte maturity. In an in vitro study, TGFß3 induced TX generation/secretion and the converting enzyme-COX-2 protein/mRNA expression both in human HO23 and primary cultured ovarian follicular GCs. While TGFßRI and Smad2/3 signaling was mainly required for COX-2 induction, ERK1/2 appeared to regulate TX secretion. The participation of Smad2/3 and COX-2 in TGFß3-induced TX generation/secretion could be further supported by the observations that Smad2/3 phosphorylation and nuclear translocation and siRNA knockdown of COX-2 expression compromised TX secretion in GCs challenged with TGFß3. Taken together, the results presented here first demonstrated that FF TGFß3 levels differ significantly in IVF patients' large preovulatory and small mid-antral follicles and are positively associated with oocyte maturation. TGFß3 can provoke TX generation by induction of COX-2 mRNA/protein via a TGFßR-related canonical Smad2/3 signaling pathway, and TX secretion possibly by ERK1/2. These imply that TGFß3 is one of the inducers for yielding FF TX in vivo, which may play a role in folliculogenesis and oocyte maturation.

Factors associated with the efficacy of mature oocyte production after dual-trigger controlled ovarian stimulation using a GnRH antagonist protocol.

BACKGROUND: The number of mature oocytes retrieved plays a significant role in determining embryo development and pregnancy outcomes of in vitro fertilization (IVF). However, studies investigating factors predictive of the efficacy of mature oocyte production (EMOP) after dual-trigger controlled ovarian stimulation (COS) are rare. This study aims to identify key predictors of EMOP during dual-trigger COS with a gonadotropin-releasing hormone (GnRH) antagonist protocol for IVF. METHODS: This retrospective cohort study included 359 first-time IVF patients undergoing dual-trigger COS with a GnRH antagonist protocol. EMOP was defined as the ratio of metaphase II (MII) oocyte count to antral follicle count (AFC). Based on EMOP results, patients were divided into two groups: group A (EMOP <70%; n = 232) and group B (EMOP ≥70%; n = 127). RESULTS: Multivariate logistic regression analysis revealed that day-2 follicle-stimulating hormone (FSH), stimulation duration, and total oocyte count were the most significant predictors of EMOP ( p < 0.05; odds ratios: 1.637, 3.400, and 1.530, respectively). Receiver operating characteristic analysis demonstrated that total oocyte count <9.5 (area under the curve [AUC], 0.782; sensitivity, 76.2%; specificity, 69.2%; p < 0.001) and stimulation duration <9.5 days (AUC, 0.725; sensitivity, 63.5%; specificity, 66.7%; p < 0.001) significantly predicted EMOP <70%. Stimulation duration combined with total oocyte count exhibited the highest power in predicting EMOP <70% (AUC, 0.767; sensitivity, 92.3%; specificity, 42.4%). CONCLUSION: Stimulation duration combined with total oocyte count was identified as the most important factor associated with the EMOP during dual-trigger COS in IVF using a GnRH antagonist protocol.

Identification of potential angiogenic biomarkers in human follicular fluid for predicting oocyte maturity.

Background: Angiogenesis in folliculogenesis contributes to oocyte developmental competence in natural and in vitro fertilization (IVF) cycles. Therefore, the identification of key angiogenic factors in follicular fluid (FF) during folliculogenesis is clinically significant and important for in vitro fertilization. This study aims to identify the key angiogenic factors in FF for predicting oocyte maturity during in vitro fertilization. Materials and methods: Forty participants who received ovarian stimulation using a GnRH antagonist protocol in their first in vitro fertilization treatment were recruited. From each patient, two follicular samples (one preovulatory follicle, > 18 mm; one mid-antral follicle, < 14 mm) were collected without flushing during oocyte retrieval. In total, 80 FF samples were collected from 40 patients. The expression profiles of angiogenesis-related proteins in FF were analyzed via Luminex high-performance assays. Recorded patient data included antral follicle count, anti-müllerian hormone, age, and BMI. Serum samples were collected on menstrual cycle day 2, the trigger day, and the day of oocyte retrieval. Hormone concentrations including day 2 FSH/LH/E2/P4, trigger day E2/LH/P4, and retrieval day E2/LH/P4 were measured by chemiluminescence assay. Results: Ten angiogenic factors were highly expressed in FF: eotaxin, Gro-α, IL-8, IP-10, MCP-1, MIG, PAI-1 (Serpin), VEGF-A, CXCL-6, and HGF. The concentrations of eotaxin, IL-8, MCP1, PAI-1, and VEGF-A were significantly higher in preovulatory follicles than those in mid-antral follicles, while the Gro-α and CXCL-6 expressional levels were lower in preovulatory than in mid-antral follicles (p < 0.05). Logistic regression and receiver operating characteristic (ROC) analysis revealed that VEGF-A, eotaxin, and CXCL-6 were the three strongest predictors of oocyte maturity. The combination of VEGF-A and CXCL-6 predicted oocyte maturity with a higher sensitivity (91.7%) and specificity (72.7%) than other combinations. Conclusion: Our findings suggest that VEGF-A, eotaxin, and CXCL-6 concentrations in FF strongly correlate with oocyte maturity from the mid-antral to preovulatory stage. The combination of VEGF-A and CXCL-6 exhibits a relatively good prediction rate of oocyte maturity during in vitro fertilization.

Trophoblast coculture induces intercellular adhesion molecule-1 expression in uterine endometrial epithelial cells through TNF-α production: Implication of role of FSH and ICAM-1 during embryo implantation.

In humans, successful implantation requires a finely tuned synchrony between an appropriately developing embryo and the receptive endometrium, involving apposition, adhesion, and invasion. Therefore, this study was sought to establish a coculture cell model to investigate trophoblast-mediated blastocyst apposition and adhesion to endometrial epithelium events during embryo implantation. The direct contact and indirect noncontact coculture models were successfully established by using human BeWo trophoblasts and HEC-1A endometrial epithelial cells. Interestingly, a significant increase of ICAM-1 protein and mRNA expression was observed in both coculture systems when challenged with follicle-stimulating factor (FSH). In accordance with these observations, trophoblast-conditioned medium (CM) could also enhance epithelial ICAM-1 production, suggesting involvement of trophoblast-secreting factor(s) in crosstalk between two cells. Indeed, FSH stimulation enhanced TNF-α expression in the trophoblasts and epithelial ICAM-1 induction were abolished by a TNF-α blocking/neutralizing antibody (TNF-α B/N Ab). Meanwhile, the intracellular calcium, PKA/CREB, and transcription/translation signaling pathways in epithelial cells participated in the ICAM-1 induction. Finally, the trophoblast cells were more susceptible to adhesion to CM-primed epithelial cell monolayer, where the adhesion could be abolished by TNF-α B/N Ab. Therefore, we present here novel findings that coculture of trophoblast with endometrial epithelial cells in the presence of FSH leads to an increase in epithelial ICAM-1 expression and trophoblast adhesion to epithelial monolayer through stimulating trophoblast's TNF-α cytokine production. This study also addresses an important issue that a possible role of microenvironmental and exogenously-added FSH in enhancing blastocyst interaction with endometrium during embryo implantation of natural or in-vitro fertilization cycle.

VEGF Concentration in a Preovulatory Leading Follicle Relates to Ovarian Reserve and Oocyte Maturation during Ovarian Stimulation with GnRH Antagonist Protocol in In Vitro Fertilization Cycle.

Serum vascular endothelial growth factor (VEGF) is involved in follicular vascularization, oxygenation, and consequently in oocyte maturation and embryo development. Unanswered questions remain regarding the relationship of intrafollicular VEGF level in preovulatory leading follicles to oocyte maturation and ovarian reserve during ovarian stimulation. We conducted this study to investigate the relationship of intrafollicular VEGF level in the fluid of single preovulatory leading follicles to ovarian reserve and oocyte maturation in patients receiving GnRH antagonist in vitro fertilization (IVF) protocol treatment. One hundred and eighty-five patients receiving IVF treatment were recruited and assigned to low-, normal-, and high-ovarian-reserve groups according to their serum anti-Müllerian hormone (AMH) level. Follicular fluid (FF) in preovulatory leading follicles, serum profiles, and clinical variables were collected for analysis. The result disclosed a significant among-group difference in FF VEGF concentration. Moreover, the serum AMH level was also negatively correlated with FF VEGF level. The oocyte maturation rate tended to be increased at higher AMH levels. FF VEGF concentration was significantly positively correlated with basal FSH level. In conclusion, FF VEGF concentration has a negative association with ovarian reserve level and oocyte maturation rate in patients undergoing GnRH antagonist IVF protocols.

TGFß1 induces in-vitro and ex-vivo angiogenesis through VEGF production in human ovarian follicular fluid-derived granulosa cells during in-vitro fertilization cycle.

A growing body of evidence indicates that angiogenesis in folliculogenesis contributes to oocyte developmental competence in natural and in-vitro fertilization (IVF) cycle of animals. Among the known angiogenic factors, vascular endothelial growth factor (VEGF) has an important role involved in angiogenesis. However, its expression level and regulatory mechanism in ovarian follicular fluid (FF) in patients undergoing IVF with controlled ovarian stimulation (COS) remains to be explored. In this study, the primary cultured human ovarian follicular granulosa cells (GCs) were prepared from FF and their identity was characterized by the presence of the GC specific markers. The transforming growth factor ß1 (TGFß1) was found to induce a significant increase in VEGF mRNA level and protein expression/secretion in GCs. In line with these observations, TGFß1 could be detected in the ovarian FF, ranging from about 400 to 2000 pg/mL among three IVF patient groups with different patient's serum Anti-Müllerian hormone level. The cellular signaling analysis revealed that TGFß1 induced VEGF production through TGFß receptor (TGFßR), Smad2/3, PI3 K/AKT, and JNK1/2-related signaling pathways. Finally, in a functional study, the TGFß1-primed GC VEGF secretion promoted in-vitro angiogenesis in vascular endothelial cells and ex-vivo vessel sprouting in aortic ring. Taken together, we demonstrated here that TGFß1 expressed in ovarian FF is an inducer for promoting VEGF production in follicular GCs through TGFßR-mediated signaling pathways and the released VEGF subsequently leads to angiogenesis. This possibly contributes to oocyte developmental competence in folliculogenesis of IVF patients with a COS protocol.

Couldn't or wouldn't? The influence of privacy concerns and self-efficacy in privacy management on privacy protection.

Sampling 515 college students, this study investigates how privacy protection, including profile visibility, self-disclosure, and friending, are influenced by privacy concerns and efficacy regarding one's own ability to manage privacy settings, a factor that researchers have yet to give a great deal of attention to in the context of social networking sites (SNSs). The results of this study indicate an inconsistency in adopting strategies to protect privacy, a disconnect from limiting profile visibility and friending to self-disclosure. More specifically, privacy concerns lead SNS users to limit their profile visibility and discourage them from expanding their network. However, they do not constrain self-disclosure. Similarly, while self-efficacy in privacy management encourages SNS users to limit their profile visibility, it facilitates self-disclosure. This suggests that if users are limiting their profile visibility and constraining their friending behaviors, it does not necessarily mean they will reduce self-disclosure on SNSs because these behaviors are predicted by different factors. In addition, the study finds an interaction effect between privacy concerns and self-efficacy in privacy management on friending. It points to the potential problem of increased risk-taking behaviors resulting from high self-efficacy in privacy management and low privacy concerns.

Problematic use of social network sites: the interactive relationship between gratifications sought and privacy concerns.

Problematic Internet use has long been a matter of concern; however, few studies extend this line of research from general Internet use to the use of social network sites (SNSs), or explicate the problematic use of SNSs by understanding what factors may enhance or reduce users' compulsive behaviors and excessive form of use on SNSs. Building on literature that found a positive relationship between gratifications sought from the Internet and problematic Internet use, this study first explores the types of gratifications sought from SNSs and examines their relationship with problematic SNS use. It found that three types of gratifications-diversion, self-presentation, and relationship building-were positively related to problematic SNS use. In addition, with a growing body of research on SNS privacy, a moderating role of privacy concerns on SNSs has been proposed to understand how it can influence the relationship between gratifications sought from SNSs and problematic SNS use. The findings suggest that different subdimensions of privacy concerns interact with gratifications sought in different manners. In other words, privacy concerns, including unauthorized secondary use and improper access, play a more influential role in constraining the positive relationship between gratifications sought and problematic SNS use when individuals seek to build relationships on SNSs. However, if individuals seek to have diversion on SNSs, their privacy concerns will be overridden by their gratifications sought, which in turn leads to problematic SNS use. Implications of these findings for future research are discussed.