Rosalina Thuffi

An increase of steroid hormones in controlled ovarian hyperstimulation (COH) procedures is reducing the success rate in assisted reproductive technology (ART), and this includes the pregnancy rate and/or implantation rate. Research has found that the decrease in the success rate occurred due to the decreased expression of the protein that is needed to prepare the endometrium so that the embryo could attach. The aim of the study was to analyse the changes in E‐chaderin expression due to COH and its relations with increased level of steroid hormones as one of the proteins in the endometrium. There were 13 samples of stored biological tissue from Macaca nemestrina endometrial tissue; came from one group of natural cycles as the control group (n = 4) and three groups of stimulated cycles. The first stimulated cycle group was injected by a 30 IU dose of rFSH (n = 2). The second stimulated cycle group was injected by a 50 IU dose of rFSH (n = 4). The third stimulated cycle group was injected by a 70 IU dose of rFSH (n = 3). The expression of E‐cadherin was measured by the immunohistochemistry (IHC) technique. Estradiol (E2) and progesterone (P4) levels were assessed using ELISA and have already been done. The IHC staining expression of E‐cadherin was found in the cytoplasm of glandular epithelium. Immunostaining measurement used the H_SCORE. We found that the expression of E‐cadherin within the group was not significantly different (p value: 0.178). Similarly, both the correlation between the estradiol level with E‐cadherin and the correlation between the progesterone level with E‐cadherin were not significantly different (p value: 0.872 and p value: 0.836). The conclusion is that the level of E‐Cadherin expression in the endometrium that were taken in themiddle secretion phase not affected by the dose regimen that given. In addition, the level of expression is not influenced by the increase of serum E2 and P4 levels.

Management of Poor Ovarian Reserve (POR) in in vitro fertilization remains a difficult challenge. The purpose of this retrospective cohort study was to compare the effectiveness of embryo banking strategy over a cohort of several mild stimulation cycles (Embryo Banking Strategy for Poor Prognosis/Embargo) to conventional full-dose antagonist protocol for IVF. Subjects identified as having poor ovarian response (POR) based on the Bologna criteria were recruited. In total, there were 113 subjects included in the analysis. Fifty-three subjects underwent embryo banking procedure (Embargo) protocol, and sixty subjects underwent the conventional full-dose antagonist protocol for IVF. The Chi-square test was used to compare the clinical pregnancy rate, miscarriage rate as well as live birth rate, while the Mann-Whitney U test was utilized to analyze the cost per clinical pregnancy between the two groups. A p<0.05 was considered statistically significant. The two studied groups showed similar outcomes regarding clinical pregnancy rate, miscarriage rate, as well as live birth rate (p=0.966, p=0.310, and p= 0.469, respectively). Cost analysis of subjects who underwent mild ovarian stimulation followed by Embargo revealed the high cost of the protocol compared to conventional full-dose antagonist protocol ($10.507±6.181 vs. $9.533±2.530, p=0.002). The clinical outcomes of both protocols were comparable. Embargo procedure was not efficient in improving the overall clinical outcomes in patients who were expected poor ovarian responders as the protocol costed more comparing with conventional full-dose antagonist protocol. A larger prospective randomized control trial is needed to evaluate this finding.

GnRH is widely used in the embryo fertilization program (FIV) as one of the ovarian stimulation regimens. At the implantation window, GnRH agonists are known to have an effect on the endometrium directly or indirectly. GnRH estimated has a negative effect on the development of endometrial cells after ovarian stimulation. This study is to analyze the impact of GnRH agonist on ovarian stimulation procedures on receptor expression and endometrial cell apoptosis. The study sample was a stored biological material (BBT) from serum and the endometrial tissue of Macaca nemestrina. The total sample is 8 and divided into 2 groups, the stimulated and control groups. Each sample will be examined 2 types which are the enzyme-linked immunosorbent assay (ELISA) for serum and immunohistochemistry (IHC) for endometrial tissue. IHC was performed with anti-GnRH and caspase 3 antibody. Concentration is measured using an ELISA reader and then converts Optical Density (OD) to a concentration using SoftMax Pro Software. Calculations of stained and unmarked cells were performed using the Image-J Analyzer programs and scored by HSCORE. There is no significant difference between GnRH serum in stimulation or control group (p=0.773). There was no correlation between GnRH in serum with GnRH receptor in endometrial tissue (p=0.060) and activation of caspase (p=0.193). There was no difference between stimulation group and control group in the amount of GnRH in serum, presence of active GnRHR, and Caspase 3 activation. Keywords: GnRH receptors, Macaca nemestria, GnRH Agonist, Controlled ovarian stimulation

Progesterone (P4) increases on the day of hCG administration in Controlled Ovarian Hyperstimulation (COH) procedure has reduced the rate of pregnancy in Assisted Reproductive Technology (ART). The aim of the study was to analyze the effect of the increase in P4 on the day of hCG administration to the expression of progesterone receptor (PR). Method: 15 samples of Macaca nemestrina endometrial tissue, in the form of stored biological materials, originated from groups of natural cycles and cycles stimulated in the COH procedure. For 16 endometrial tissue samples, PR expression was measured by immunohistochemical (CPI) technique. The level of PR expression was compared between the natural cycle and the stimulated cycle that got P4≥1 ng/mL and P4 0.05). Conclusion, the level of P4 > 1 ng / mL on the day of administration of hCG in the COH procedure decreases PR expression in the endometrial gland.

The increase in progesterone (P4) levels on the day of human chorionic gonadotropin (hCG) administration have a negative effect on endometrial receptivity. There are few reports regarding the expression of homeobox A10 (HOXA10) as one of many biomolecular factors of endometrial receptivity. To evaluate the effect of increased P4 concentration on the day of hCG administration on HOXA10, a total of 16 Macaca nemestrina were divided into three dose groups of recombinant-follicle stimulating hormone (rFSH) (30IU, 50IU, and 70IU) and one control group. Injection of rFSH combined with gonadotropin release hormone (GnRH) at 160 ug/day was given subcutaneously using a long protocol technique. Blood samples for estradiol (E2) and (P4) concentration measurements were taken on the day of injecting hCG in the final follicular phase, while the collection of endometrial tissue for HOXA10 measurement was carried out 8 to 10 days after hCG administration. E2 and P4 were measured by ELISA, whereas HOXA10 expression was measured with immunohistochemical (IHC) techniques. The concentration of E2 and P4 was found to be higher in dose groups compared with the natural group, but no significant differences were found within the group. For the Hscore for HOXA10 expression, no significant differences within dose groups were found. In addition, no significant differences for the Hscore for HOXA10 were found when compared to E2 groups. Significantly, the Hscore of HOXA10 was found to be >1 ng/mL in the P4 group compared with the Hscore HOXA10 in the P4 natural group (p = 0.022). The high concentration of P4 caused by ovarian hyperstimulation in the follicular phase stimulates the expression of HOXA10 in the secretion phase.

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