Comparative Effects of Metformin and Dienogest on PI3K/AKT Signaling in Experimental Endometriosis

Comparative Effects of Metformin and Dienogest on PI3K/AKT Signaling in Experimental Endometriosis | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Comparative Effects of Metformin and Dienogest on PI3K/AKT Signaling in Experimental Endometriosis Seyedmahdi Tabatabaei, Nazlı Çil, Elif Önder, İsmail Kubilay Baykan, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-10557372/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This study investigates the therapeutic effects of Metformin and Dienogest on the PI3K/AKT signaling pathway in a rat model of induced endometriosis, aiming to enhance treatment strategies. Experimental groups included untreated control rats, rats with surgically induced endometriosis, and treatment groups receiving Dienogest (0.3 mg/kg/day) and Metformin (100 mg/kg/day) for 28 days. Treatments began on the 21st day post-induction and continued until sacrifice on day 49. Uterine cycle was monitored via vaginal smears, and endometrial lesion areas were reassessed pre- and post-treatment. Blood estradiol (E2), progesterone, and VEGF levels were evaluated using enzyme-linked immunosorbent assay (ELISA). The endometriotic implants and uterus were assessed using histopathological analysis. The expression of PI3K/AKT pathway proteins was evaluated in endometriosis lesions and uterine tissue using immunohistochemical and RT-PCR methods. Metformin significantly reduced lesion diameter compared with untreated and dienogest-treated groups (p = 0.002). Serum E2 levels were significantly higher in the metformin group (p = 0.05). Histological examination showed reduced inflammatory infiltration and tissue degeneration in both treatment groups. RT-PCR demonstrated that PI3K, p-PI3K, and AKT expression were increased in the treatment groups within the endometriosis lesions. However, PI3K and p-PI3K expression were found to be reduced in uterine tissue. AKT expression was statistically significantly higher in the metformin group (p = 0.05). These findings confirm Metformin and Dienogest as effective agents for endometriosis regression, with molecular-level differences suggesting distinct mechanisms of action. Metformin's comprehensive effects on macroscopic, biochemical, microscopic, and molecular levels underscore its therapeutic potential. Further research is essential to unravel its mechanisms fully and explore clinical applications. Endometriosis PI3K AKT Metformin Dienogest Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Endometriosis is an estrogen-dependent disorder that affects approximately 6–10% of women in their reproductive years (1). Characterized by pelvic pain, dysmenorrhea, dyspareunia, and infertility, its manifestations can extend to systemic inflammation and immune irregularities, increasing the risk of cardiovascular diseases, rheumatoid arthritis, and even certain cancers like ovarian and breast cancer (2). Despite its significant prevalence, the underlying mechanisms of endometriosis remain poorly understood, with its pathogenesis linked to hormonal imbalance, chronic inflammation, and metabolic alterations (1, 3). Endometriosis is characterized by the ectopic growth of endometrial tissue outside the uterus, most frequently involving the ovaries, peritoneum, and pelvic cavity (3). Conventional treatment approaches include surgical excision of endometriotic lesions followed by hormonal suppression, yet these methods often yield suboptimal outcomes with risks of recurrence and significant adverse effects (4). Consequently, there is a need to explore novel therapeutic strategies that address the molecular pathophysiology of the disease. One promising area of research is the role of the PI3K/AKT signaling pathway in the progression and management of endometriosis. This pathway plays an important role in regulating cell growth, survival, metabolism, and angiogenesis, processes that are aberrantly activated in endometriotic tissues (5). Previous studies have highlighted the therapeutic potential of targeting this pathway in various diseases, including cancer and chronic inflammatory disorders (6, 7). This study focuses on assessing the effects of Metformin and Dienogest, two pharmacological agents, on the PI3K/AKT pathway in a rat model of induced endometriosis. By investigating these molecular mechanisms, this research aims to contribute to the development of targeted treatments that not only alleviate the symptoms of endometriosis but also address its underlying pathophysiological processes. Materials and Methods Creation of an Animal Model of Endometriosis This study was approved by the Pamukkale University Animal Experiments Ethics Committee (PAUHADYEK-2023-07). Thirty-two adult female Sprague Dawley rats were used. During the estrus phase, brief anesthesia was administered, followed by a 5-cm midline abdominal incision. One uterine horn was ligated and excised at both the uterotubal junction and cervical end. The excised horn was immersed in sterile saline, the endometrium was exposed by dividing along the antimesenteric axis, and 5 × 5 mm sections were prepared. These explants were sutured to the omentum with 7 − 0 Vicryl, ensuring the endometrial surface faced the peritoneum or mesentery. Each segment was fixed at four corners, and the abdominal layers were anatomically closed with 4 − 0 Vicryl. Animals were allowed to recover from anesthesia (8). The rats with experimental endometriosis were randomly divided into 3 groups 21 days after the surgical procedure. The animals were treated daily as follows: Control Group (n: 8): Group that received nothing. Endometriosis Group (n: 8): Group in which experimental endometriosis was induced. Endometriosis+Dionegest Group (n:8): Group in which experimental endometriosis was induced and Dienogest was administered orally at 0.3 mg/kg/day for 28 days. Endometriosis+Metformin Group(n: 8): Group in which experimental endometriosis was created, and Metformin was given orally at 100 mg/kg/day. All doses were administered orally from day 21 to day 49. After day 49, vaginal smears were taken, and rats were sacrificed after the diestrous period was detected (8). Subsequent to a re-evaluation of the endometrial lesion area, the left uterine horn was excised. The results regarding the endometrial lesion area were measured by the surface area (length x width x height in millimeters) in each group. After the tissues were fixed in formaldehyde, sections were taken after routine tissue follow-up, hematoxylin eosin was stained and evaluated. PI3K, p-PI3K, AKT, and p-AKT reactions in the tissues were evaluated immunohistochemically. Some of the uterus and endometriosis tissues were separated and stored in Trizol at -80°C. Following the isolation of total RNA from the tissues, alterations in mRNA expression were analysed using reverse transcription-polymerase chain reaction (RT-PCR). Vaginal Smear Application 28 days after the experimental procedure was established, vaginal smears were performed for 32 rats every day at 10:00, and estrus cycles were observed. Rats with dense, fragmented leukocyte dominance as a result of the smear were evaluated as being in the diestrus phase and were sacrificed (8)(Fig. 1 ). Elisa Method The collection of blood samples was performed by intracardiac puncture in rats under anaesthesia, and the samples were subsequently labelled for identification. The samples were then subjected to an incubation period at ambient temperature for a duration of one hour. This was followed by a centrifugal process at 400 g for a period of 15 minutes, with the objective of effecting a separation of the upper layer from the sample. The sera obtained were then aliquoted into appropriately labelled tubes and stored within a temperature of -80°C, pending further analysis. On the day of the study, the samples were allowed to reach room temperature prior to testing. The quantification of progesterone, estradiol (E2) and VEGF was conducted by means of enzyme-linked immunosorbent assay (ELISA) kits (BT LAB) in accordance with the manufacturer's protocols. Absorbance readings at 450 nanometers were performed using a Biotek Elx800 Microplate Reader (BioTek Instruments Inc., USA), and serum concentrations were calculated using the Gen5 data analysis software. Real-Time Polymerase Chain Reaction (RT-PCR) PCR was performed with the Thermo Scientific PikoReal 96 device, which can read 96-well microplates. PI3K, p-PI3K and AKT expressions were determined using RT-PCR. Total RNA was isolated from the relevant tissue using TRI-Reagent, and the quantity and quality of the obtained RNAs were subsequently determined. Afterwards, cDNA synthesis was performed from total RNAs, and expression analysis was performed with reference-housekeeping B-actin RT-PCR (Table 1 ). Histopathological Examination Ectopic endometrial and uterine tissues were fixed in 10% formalin for 72 hours and processed using standard paraffin-embedding techniques. Sections measuring 5 µm in thickness were prepared, deparaffinized in xylene, rehydrated through graded ethanol, and rinsed in distilled water. The samples were subjected to a staining process involving hematoxylin for a duration of three minutes, followed by a thorough wash. To differentiate the samples, an acid alcohol solution was employed. Following the application of a bluing solution, the sections were counterstained with eosin for a period of three minutes. The process of dehydration was conducted through the use of graded ethanol, following which the sections were cleared in xylene and mounted with Entellan for microscopic evaluation. Immunohistochemical Staining Method Immunohistochemical staining was conducted to assess protein expressions of PI3K, p-PI3K, AKT, and p-AKT using specific primary antibodies (FineTest). Antigen retrieval was carried out using a trypsin solution (0.2 g in 200 mL PBS) at 37°C for 30 minutes. Endogenous peroxidase activity was blocked using a 3% hydrogen peroxide-methanol solution. Primary antibodies were incubated overnight at 4°C at appropriate dilutions: PI3K (1:50), p-PI3K (1:50), AKT (1:200), and p-AKT (1:200). Following secondary antibody application and streptavidin-HRP treatment, sections were visualized using 3,3′-diaminobenzidine as the chromogen. Counterstaining with hematoxylin enhanced visual clarity. The staining intensity was evaluated semi-quantitatively using an H-score method, categorizing reactions as strong (+++), moderate (++), weak (+), or absent (-). Statistical Analysis The data were analysed using SPSS version 25.0 (IBM Corp., Armonk, NY, USA). Continuous variables were presented as mean ± standard deviation, median with interquartile range (25th–75th percentiles), and minimum–maximum values, while categorical variables were expressed as frequencies and percentages. The normality of data distribution was assessed using the Shapiro–Wilk test. In instances where a comparison between groups was required, a One-Way ANOVA followed by a Tukey's post hoc test was employed, provided that the parametric assumptions were met. In instances where the initial assumptions were not met, the Kruskal–Wallis test was employed, with Mann–Whitney U tests and Bonferroni correction for post hoc analysis. Paired comparisons were conducted using either the paired samples t-test (parametric) or the Wilcoxon signed-rank test (non-parametric). A p-value of less than 0.05 was considered to be statistically significant in all analyses. Results Animal Weight Findings When the initial and final weights of the patients were compared, no significant difference was found between the groups (p > 0.05). ELISA Findings Serum estradiol (E2) concentrations were significantly elevated in the Endometriosis + Metformin group compared with all other groups. Despite the absence of statistical significance in the differences observed in progesterone concentrations among the groups, the highest values were recorded in the metformin-treated rats. Levels of vascular endothelial growth factor (VEGF) demonstrated no significant variation between groups; however, the lowest mean value was observed in the control group, whereas the highest was detected in the endometriosis + metformin group (44.53 ± 30.2 vs. 86.93 ± 42.2) (Table 2 ). Table 2 Statistical comparison of blood hormone levels of groups Control Endometriosis Endometriosis+Dienogest Endometriosis+Metformin p E2(pg/mL) A.O \(\:\pm\:\) S.S 91.1 ± 52.09 139.77 ± 178.67 74.35 ± 19.08 308.65 ± 152.94 0.05* (kw = 7.741) Med(IQR) 95.83(40.89-141.62) 95.83(40.89-141.62) 81.25(54.67–87.13) 349.3(164.34-435.91) * Progestron(pg/mL) A.O \(\:\pm\:\) S.S 113.35 ± 12.68 146.71 ± 106.03 170.01 ± 77.73 199.99 ± 35.89 0.139(kw = 5.487) Med(IQR) 108.51(101.38-124.15) 113.55(74.11-222.93) 167.36(99.8-242.88) 210.48(195.06-220.03) VEGF(pg/mL) A.O \(\:\pm\:\) S.S 44.53 ± 30.2 68.75 ± 64.37 54.28 ± 44.74 86.93 ± 42.2 0.577(F = 0.675) Med(IQR) 56.46(12.98–64.15) 54.16(18.01-111.49 56(11-95.83) 70.02(61.01-109.54) *p < 0.05 statistically significant difference; A.O: Arithmetic mean; SD: Standard Deviation; Med (IQR): Median (25th − 75th Percentiles); kw: Kruskal Wallis Variance Analysis Endometriosis Macroscopy Findings On the 28th day, a comparison of the lesion diameters among the endometriosis groups revealed significant differences. The Endometriosis + Metformin group demonstrated the most minimal lesions (3.5 ± 2.36), which were considerably lower than those observed in the untreated Endometriosis group (177.8 ± 214.01) and the Endometriosis + Dienogest group (103 ± 143.58) (p = 0.002). Despite the observation that lesion size in the Dienogest group was reduced in comparison with the untreated group, this difference did not attain statistical significance (Fig. 2 ). RT-PCR Findings When endometriosis foci were compared, p-PI3K expression showed a statistically significant increase in the Endometriosis+Dienogest group compared to the Endometriosis groups. It was determined that p-PI3K expression increased 2.96 times. When PI3K expression was compared to the Endometriosis group, a statistically significant increase was detected in the Endometriosis+Dienogest group, and the expression of PI3K was found to be 1.88 times higher. When endometriosis foci and the Endometriosis+Metformin group were compared, no statistically significant difference was detected in PI3K, AKT and p-PI3K expression. Although the expression of these antibodies was found to be higher in the Endometriosis+Metformin group (Table 3 ). Table 3 Changes in gene expression levels in endometriosis foci of the groups according to RT-PCR results Gen Symbol Fold Regulation (compared with the control group) Endometriosis+Dienogest Endometriosis+Metformin Fold Regulation p-value Fold Regulation p-value Beta-actin 1.00 nan 1.00 nan PI3K 1.88 0.002* 4.79 0.24 p-PI3K 2.96 0.01* 2.70 0.36 AKT 1.13 0.40 4.41 0.72 *p < 0.05 statistically significant difference When the expressions of the uterine tissues of the groups were compared, p-PI3K expression in the Endometriosis group decreased statistically significantly compared to the control group. PI3K expression was significantly decreased in the uterine tissue of the Endometriosis+Dienogest group compared to the control group. When the protein expressions of the Endometriosis+Metformin group were compared to the control group, it was found that AKT expression increased statistically significantly in the uterine tissue (Table 4 ). Table 4. Changes in gene expression levels in the uterine tissue of control and other groups according to RT-PCR results. Gen Symbol Fold Regulation (compared with the control group) Endometriosis Endometriosis+Dienogest Endometriosis+Metformin Fold Regulation p-value Fold Regulation p-value Fold Regulation p-value Beta-actin 1.00 nan 1.00 nan 1.00 nan *PI3K -2.63 0.85 -21.64 0.01* -11.80 0.15 p-PI3K -4.97 0.0002* -11.47 0.08 -9.63 0.15 AKT -2.40 0.28 2.03 0.24 73.86 0.05* *p<0.05 statistically significant difference Hematoxylin & Eosin Staining Findings In the Endometriosis group, where ectopic endometrial tissue was observed, stromal cells were widespread in the tissue, and there was an accumulation of inflammatory cells and numerous vascularizations. In some areas, there was epithelial tissue consisting of small cells and structures similar to a glandular structure surrounded by cells. The collagen fibers, in the form of thick bundles forming a network with each other in the cellular area, were remarkable. It was observed that ectopic endometrial tissue was relatively smaller in the Endometriosis+Dienogest and Endometriosis+Metformin groups, and the tissue consisting of stromal and inflammatory cells remained in a very small area (Fig. 3 ). In the control group of uterus tissue, endometrium, myometrium, and perimetrium layers were observed to be normal structure. In the endometriosis group, it was observed that the surface epithelium lost its continuity and was interrupted in places. It was determined that there were openings in the stroma and that the glands were separated from it. Some of the glands had lost their normal structure and were curled. It was also noted that the muscle fibres in the myometrium layer had lost their integrity and had separated, forming gaps between them and the endometrium. It was observed that the degeneration decreased and the integrity was preserved in the uterine in the Endometriosis+Dienogest group. However, there were still interruptions in the epithelium, separation of the glands from the stroma, and gaps in the stromal tissue. The myometrium and perimetrium layers were observed to be normal. The Endometriosis+Metformin group was similar to the Endometriosis+Dienogest group. It was observed that degenerative changes in the endometriosis group decreased. In this group, there were still some interruptions in the epithelium lining the lumen and separation of the glands from the stromal tissue. Myometrium and perimetrium layers were evident (Fig. 3 ). Immunohistochemical Findings Endometriosis Lesion Immunohistochemical Findings PI3K expression was determined to be higher in the Endometriosis+Metformin group compared to the other groups, while p-PI3K was lower. AKT expression was higher in the Endometriosis+Dienogest group compared to the Endometriosis (p = 0.004) and Endometriosis+Metformin (p = 0.025) groups. p-AKT expression was found to be higher in the Endometriosis+Dienogest and Endometriosis+Metformin group compared to the Endometriosis group (Fig. 4 ). Uterus Immunohistochemical Findings PI3K expression was higher in the control group compared to the other groups (p = 0.004). Although the expression was higher in the Endometriosis group compared to the Endometriosis+Dienogest group (p = 0.004), it was not significant compared to the Endometriosis+Metformin group. Expression was strong in the epithelium and some areas of the stroma in the control group. p-PI3K expression was higher in the Control, Endometriosis+Dienogest, and Endometriosis+Metformin groups compared to the Endometriosis group (p = 0.004). The expression distribution was strong in the epithelium, glands, stroma, outer layer of the myometrium, and perimetrium in both the Endometriosis+Dienogest and Endometriosis+Metformin groups. AKT expression was higher in the Endometriosis group compared to the other groups (0.004). Expression was higher in the Endometriosis+Metformin group than in the Endometriosis+Dienogest group (p = 0.002). The p-AKT expression of the Control and Endometriosis groups was higher than the Endometriosis+Dienogest and Endometriosis+Metformin groups (p = 0.004). Expression was weak in the epithelium in the control group, strong in the glands, and moderate in the stroma; however, the inner layer of the myometrium was weak, the outer layer and the perimetrium were dense. In the Endometriosis group, the epithelium layer was weak, the gland and stroma were moderate, and the myometrium and perimetrium layers showed moderate reaction. In the Endometriosis+Dienogest group, the epithelium layer, stroma, and myometrium layer showed weak to moderate staining, while the glands and perimetrium layer showed intense staining. In the Endometriosis+Metformin group, a moderate reaction was observed in the epithelium, stroma, and myometrium, and a weak reaction in the glands (Fig. 5 ). Discussion In our study, the treatments provided improvement in endometriosis foci. The most significant improvement was found in the Endometriosis+Metformin group. Interestingly, the Endometriosis+Metformin group exhibited significantly higher serum E2 levels despite demonstrating the greatest lesion regression, suggesting that its anti-endometriotic activity may not be directly mediated through estrogen suppression. Dienogest and metformin increased activation of the PI3K-AKT signaling pathway in endometriosis foci. While RT-PCR findings showed significant upregulation of PI3K and p-PI3K expression in the dienogest group, immunohistochemical analyses demonstrated differential regulation of PI3K, p-PI3K, AKT, and p-AKT proteins in both treatment groups. These findings suggest that metformin and dienogest may exert their beneficial effects through distinct mechanisms involving modulation of PI3K/AKT signaling. Dienogest has been shown in multiple clinical trials to reduce endometriotic lesions and relieve pain effectively. Furthermore, its safety profile is favourable, characterised by the absence of androgenic effects and only minimal suppression of ovarian function (9–12). In one experimental study, rats with surgically induced endometriosis received oral dienogest at doses of 0.3 or 1.0 mg/kg/day for a period of 28 days. Treatment with 0.3 mg/kg/day led to a significant reduction in the total endometrial lesion area, demonstrating an efficacy level comparable to that of 100 mg/kg/day of danazol. In contrast to the higher dose, dienogest 0.3 mg/kg/day did not result in alterations to uterine horn weight, indicating the absence of estrogenic effects at this level in rodents. The model involved the transplantation of endometrial-like tissue at two peritoneal sites (peritoneum and mesenterium). Following exposure to 0.3 mg/kg/day dienogest, the expression of vascular endothelial growth factor (VEGF) mRNA was measured in lesion tissue, and no significant differences were observed in comparison with the control group (13). Katayama et al. demonstrated that the oral administration of dienogest for a period of up to 14 days was capable of suppressing angiogenesis in an autologous rat model of endometriosis. The authors of the study hypothesised that this effect may be attributable to antiestrogenic activity or direct progestin-mediated regulation of VEGF expression (14). In our study, E2 and VEGF levels were lower in the Endometriosis+Dienogest group than in the untreated Endometriosis group, although the differences were not statistically significant. This suggests that the beneficial effects of dienogest may be related to local anti-proliferative and anti-inflammatory actions rather than changes in circulating E2 or VEGF levels. In contrast, metformin treatment significantly increased serum E2 levels. However, circulating estrogen levels may not reflect the local hormonal environment of endometriotic lesions. Therefore, the effects of Metformin and Dienogest are likely mediated through local mechanisms independent of systemic E2 and VEGF changes. An in vitro investigation demonstrated that metformin inhibited the inflammatory response, suppressed aromatase activation, and reduced proliferation in cultured endometriotic stromal cells under sterile conditions (15). Moreover, metformin has been demonstrated to exert an antiproliferative effect on endometrial glands (16). Furthermore, the presence of aromatase expression in endometriotic tissues was accompanied by elevated local estrogen concentrations, indicating that estradiol was produced in situ through aromatase activity (17). In addition, metformin has been reported to suppress follicle-stimulating hormone (FSH) activity and to inhibit insulin-stimulated production of progesterone and estradiol in granulosa cells (18). Consequently, the hypothesis has been put forward that metformin may play a role in the prevention of endometriosis through the suppression of both ovarian and local estrogen production. Supporting evidence comes from an in vitro study in which stromal cells derived from human ovarian endometrioma were cultured and exposed to metformin for 24 hours. Metformin has been demonstrated to exert a substantial anti-inflammatory effect, whilst concomitantly attenuating estradiol production and the proliferation of endometriotic stromal cells. In addition, metformin has been shown to inhibit cAMP-stimulated aromatase activity in endometriotic lesions, with this effect being dose-dependent (Takemura et al., 2007; Oner et al., 2010). Beyond the scope of endometriosis, metformin has been advanced as a therapeutic agent for ERα-positive breast cancer, a proposition that can be attributed to its capacity to suppress ERα expression and function (19). In another study, 27-week-old female mice were divided into control and metformin-treated groups. The metformin group was fed a diet containing 100 mg/kg metformin for six months. Compared with controls, metformin mice had higher serum estradiol (E2) levels and regular estrous cycles, while follicle-stimulating hormone (FSH) levels remained unchanged. These findings suggest that metformin may enhance ovarian function in middle-aged animals (20). In the present study, serum estradiol (E2) levels were found to be significantly elevated in the Endometriosis + Metformin group. Although metformin has previously been reported to suppress local estrogen production within endometriotic lesions, the present findings suggest that this effect may not necessarily translate into reduced circulating estrogen concentrations. Conversely, the reduced E2 levels observed in the Dienogest-treated group are consistent with the established endocrine actions of Dienogest described in earlier studies. A study assessing oral treatments with metformin (100 mg/kg/day or 200 mg/kg/day) and letrozole (0.1 mg/kg/day) in a rat model of endometriosis found that the lowest histopathological implant scores occurred with 100 mg/kg/day metformin. The administration of metformin in this dosage resulted in the regression of endometriotic implants and a reduction in the severity of adhesions (21). In accordance with the findings of earlier research, adhesions were also observed at the sites of endometrial implants in the present study. The lowest degree of adhesion formation was observed in the Endometriosis + Metformin group. This reduction may be attributed to the anti-inflammatory and antiproliferative properties of metformin, which likely contribute to limiting adhesion development. A study was conducted in which tissue was excised from rats during the diestrus phase in order to examine endometriosis-induced tissue. The results of the histological evaluation revealed the presence of ectopic endometrium, which was characterised by pseudostratified epithelium and vacuolated columnar cells alternating with darker cells. The surrounding stroma appeared edematous and contained dilated, congested blood vessels (22). In a study evaluating the effects of cisplatin and letrozole on surgically induced endometriosis, both treatments significantly reduced implant growth and histological scores in comparison with the untreated control group (23). Another experimental study demonstrated that ornidazole treatment limited the progression of endometriosis in a rat model through its anti-inflammatory and anti-angiogenic properties. After 14 days of therapy, lesion size was significantly reduced in the ornidazole group compared with controls. Histological evaluation with hematoxylin and eosin staining revealed endometrial atrophy, thinning of the epithelial layer, and decreased glandular tissue in treated rats. Furthermore, both microvessel density and the expression of IL-1β, IL-6, TNF-α, and VEGF within endometriotic lesions were markedly lower following ornidazole administration (24). In a study by Ozdemir et al., a morphological evaluation of endometrial implants was conducted, which revealed increased cystic changes and vascularity, accompanied by stromal inflammation, capillarization, and vascular engorgement. The endometrial epithelium and glands were more distinctly identified in these lesions. A morphometric analysis was conducted to compare the endometriotic foci in the zoledronic acid- and ranibizumab-treated groups with those in the control and other treatment groups. The results showed that the foci in the zoledronic acid- and ranibizumab-treated groups appeared morphologically reduced and embedded within the tissue. The aforementioned groups also exhibited decreased epithelial integrity, reduced gland density, stromal degeneration, diminished inflammation, lower capillary activity, and reduced vascular filling. A statistically significant difference was observed among groups in the distribution of epithelial histological scores (p < 0.001), with scores in the zoledronic acid and ranibizumab groups significantly lower than those in the control, dienogest, danazol, and buserelin acetate groups (p < 0.001) (25). In the present study, ectopic endometrial tissue in the endometriosis group was characterised by abundant stromal cells, prominent inflammatory cell infiltration, and numerous blood vessels. In specific regions, the presence of epithelial tissue, characterised by low-lying cells and gland-like structures, was identified, with these structures being surrounded by stromal elements. Additionally, the presence of thick bundles of collagen fibres, forming interconnected networks within the cellular space, was also notable. In the uterine tissue, the surface epithelium of the endometriosis group exhibited loss of continuity, with interruptions in several regions. Conversely, in the Endometriosis + Dienogest and Endometriosis + Metformin groups, ectopic endometrial tissue exhibited a significant reduction, with stromal and inflammatory cells restricted to a confined area. The histological improvements observed in response to metformin treatment may be attributable to its anti-inflammatory, antiproliferative, and anti-angiogenic properties, as reported in previous studies. In a similar manner, the advantageous effects of dienogest are presumably derived from its capacity to impede endometrial proliferation and elicit regression of endometriotic lesions. Collectively, these findings suggest that both metformin and dienogest have therapeutic potential in mitigating endometriosis-associated histopathological changes and limiting disease progression. In a study investigating PI3K/AKT pathway regulation in human endometrial biopsies, ectopic tissue samples from 20 women with untreated peritoneal endometriosis and endometrial biopsies from 9 controls were analysed. The results demonstrated increased PI3K expression, reduced PTEN expression, and elevated AKT phosphorylation in ectopic endometrium compared with controls. It is noteworthy that levels of PTEN were found to be significantly lower in both ectopic tissue and eutopic endometrium in patients with minimal to mild disease compared with those with moderate to severe endometriosis. Furthermore, the p-AKT/total AKT ratio indicated higher AKT phosphorylation in endometriotic tissue from patients with minimal to mild disease. Collectively, these findings confirm dysregulation of the PI3K/AKT signaling pathway in endometriosis (7). In our study, PCR and immunohistochemical findings revealed that PI3K/AKT signalling pathway expression did not decrease in endometriosis foci within the treatment groups compared to the endometriosis group. Notably, despite reduced lesion size and histopathological improvement, there was no decrease in PI3K/AKT pathway expression. PI3K expression was highest in the control group in uterine tissue. According to the PCR results, p-PI3K expression was also highest in the control group. However, according to the immunohistochemical results, the control group was similar to the treatment groups, with the lowest expression observed in the endometriosis group. AKT expression decreased in the Endometriosis+Dienogest group; however, it was statistically significantly higher in the Endometriosis+Metformin and Endometriosis groups. These findings suggest that the histopathological improvements observed after metformin and dienogest treatment may not be solely explained by modulation of the PI3K/AKT pathway. Our study has certain limitations. Only a single dose and treatment duration were examined for metformin and dienogest; therefore, dose-dependent and long-term effects could not be assessed. Functional outcomes related to fertility and pain—which are clinically significant symptoms of endometriosis—were not evaluated in this experimental model. Conclusion In conclusion, Metformin and Dienogest caused regression of implants in the rat model of endometriosis. However, their effects on protein expression related to proliferation and angiogenesis, as well as on serum sex hormone levels, differed. Metformin was identified as the most effective treatment at macroscopic, microscopic, and molecular levels. Further experimental and clinical studies are needed to understand the mechanism of action of Metformin better. Declarations Author Contributions: N.Ç., S.T., K.B., and A.C. were involved in the design of the experiments and drafted the manuscript. N.Ç., S.T., H.Ş., and G.A.M. were involved in data analysis. G.A.M., and N.Ç. reviewed and edited the manuscript. All authors read and approved the final version of the manuscript. Funding: This study was supported by the Pamukkale University Scientific Research Projects Coordination Unit (PAUBAP) under project number PAUHADYEK-2023-07, approved by the Pamukkale University Animal Experiments Ethics Committee. Conflict of Interest: The authors declare no conflict of interest. Ethics Approval : All experimental procedures involving animals were reviewed and approved by the Pamukkale University Animal Experiments Ethics Committee (PAUHADYEK) under approval number PAUHADYEK-2023-07. References Zondervan KT, Becker CM, Missmer SA. Endometriosis. N Engl J Med. 2020;382(13):1244-56. Kvaskoff M, Mu F, Terry KL, Harris HR, Poole EM, Farland L, et al. Endometriosis: a high-risk population for major chronic diseases? Human reproduction update. 2015;21(4):500-16. Horne AW, Saunders PT, Abokhrais IM, Hogg L. Top ten endometriosis research priorities in the UK and Ireland. The Lancet. 2017;389(10085):2191-2. Saraswat L, Ayansina D, Cooper K, Bhattacharya S, Horne A, Bhattacharya S. Impact of endometriosis on risk of further gynaecological surgery and cancer: a national cohort study. BJOG: An International Journal of Obstetrics & Gynaecology. 2018;125(1):64-72. He Y, Sun MM, Zhang GG, Yang J, Chen KS, Xu WW, et al. Targeting PI3K/Akt signal transduction for cancer therapy. Signal transduction and targeted therapy. 2021;6(1):425. Gentilini D, Busacca M, Di Francesco S, Vignali M, Vigano P, Di Blasio A. PI3K/Akt and ERK1/2 signalling pathways are involved in endometrial cell migration induced by 17β-estradiol and growth factors. Molecular human reproduction. 2007;13(5):317-22. Madanes D, Bilotas MA, Bastón JI, Singla JJ, Meresman GF, Barañao RI, et al. PI3K/AKT pathway is altered in the endometriosis patient’s endometrium and presents differences according to severity stage. Gynecological Endocrinology. 2020;36(5):436-40. Oner G, Ozcelik B, Ozgun MT, Serin IS, Ozturk F, Basbug M. The effects of metformin and letrozole on endometriosis and comparison of the two treatment agents in a rat model. Human reproduction. 2010;25(4):932-7. Köhler G, Faustmann TA, Gerlinger C, Seitz C, Mueck AO. A dose-ranging study to determine the efficacy and safety of 1, 2, and 4 mg of dienogest daily for endometriosis. International Journal of Gynecology & Obstetrics. 2010;108(1):21-5. Seitz C, Gerlinger C, Faustmann T, Strowitzki T. Safety of dienogest in the long-term treatment of endometriosis: a one-year, open-label, follow-up study. Fertility and Sterility. 2009;92(3):S107. Strowitzki T, Faustmann T, Gerlinger C, Seitz C. Dienogest in the treatment of endometriosis-associated pelvic pain: a 12-week, randomized, double-blind, placebo-controlled study. European Journal of Obstetrics & Gynecology and Reproductive Biology. 2010;151(2):193-8. Strowitzki T, Marr J, Gerlinger C, Faustmann T, Seitz C. Dienogest is as effective as leuprolide acetate in treating the painful symptoms of endometriosis: a 24-week, randomized, multicentre, open-label trial. Human Reproduction. 2010;25(3):633-41. Fischer O, Kaufmann-Reiche U, Moeller C, Fuhrmann U. Effects of dienogest on surgically induced endometriosis in rats after repeated oral administration. Gynecologic and obstetric investigation. 2011;72(3):145-51. Katayama H, Katayama T, Uematsu K, Hiratsuka M, Kiyomura M, Shimizu Y, et al. Effect of dienogest administration on angiogenesis and hemodynamics in a rat endometrial autograft model. Human reproduction. 2010;25(11):2851-8. Takemura Y, Osuga Y, Yoshino O, Hasegawa A, Hirata T, Hirota Y, et al. Metformin suppresses interleukin (IL)-1β-induced IL-8 production, aromatase activation, and proliferation of endometriotic stromal cells. The Journal of Clinical Endocrinology & Metabolism. 2007;92(8):3213-8. Li L, Mamputu J-C, Wiernsperger N, Renier G. Signaling pathways involved in human vascular smooth muscle cell proliferation and matrix metalloproteinase-2 expression induced by leptin: inhibitory effect of metformin. Diabetes. 2005;54(7):2227-34. Attar E, Bulun S. Aromatase and other steroidogenic genes in endometriosis: translational aspects. Human reproduction update. 2006;12(1):49-56. Mansfield R, Galea R, Brincat M, Hole D, Mason H. Metformin has direct effects on human ovarian steroidogenesis. Fertility and sterility. 2003;79(4):956-62. Kim J, Lee J, Jang SY, Kim C, Choi Y, Kim A. Anticancer effect of metformin on estrogen receptor-positive and tamoxifen-resistant breast cancer cell lines. Oncology reports. 2016;35(5):2553-60. Qin X, Du D, Chen Q, Wu M, Wu T, Wen J, et al. Metformin prevents murine ovarian aging. Aging (Albany NY). 2019;11(11):3785. Cheng J, Li C, Ying Y, Lv J, Qu X, McGowan E, et al. Metformin alleviates endometriosis and potentiates endometrial receptivity via decreasing VEGF and MMP9 and increasing leukemia inhibitor factor and HOXA10. Frontiers in pharmacology. 2022;13:750208. Elgamal DA, Othman E-ER, Ahmed SF. Ultrastructural features of eutopic endometrium in a rat model of endometriosis. Journal of Microscopy and Ultrastructure. 2016;4(1):20-7. Li Z, Liu H, He Z, Zhang G, Lang J. Effects of cisplatin and letrozole on surgically induced endometriosis and comparison of the two medications in a rat model. European Journal of Pharmaceutical Sciences. 2016;93:132-40. Qin X, Yang H, Qiao D, Liu X, Liu L-B, Liu S-F, et al. Ornidazole Reduces the progression of endometriosis in a rat model. Gynecologic and Obstetric Investigation. 2022;87(5):316-23. Ureyen Ozdemir E, Adali E, Islimye Taskin M, Yavasoglu A, Aktug H, Oltulu F, et al. Effects of ranibizumab and zoledronic acid on endometriosis in a rat model. Archives of gynecology and obstetrics. 2022;305(1):267-74. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-10557372","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":693531614,"identity":"9ae515db-f136-471a-a777-c19b77d3e21b","order_by":0,"name":"Seyedmahdi Tabatabaei","email":"data:image/png;base64,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","orcid":"","institution":"Pamukkale University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Seyedmahdi","middleName":"","lastName":"Tabatabaei","suffix":""},{"id":693531616,"identity":"18b93f43-06a5-447e-be6d-3157f98fbc6e","order_by":1,"name":"Nazlı Çil","email":"","orcid":"","institution":"Pamukkale University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Nazlı","middleName":"","lastName":"Çil","suffix":""},{"id":693531617,"identity":"2a4c0111-caa9-4d4b-aebd-905caf417f45","order_by":2,"name":"Elif Önder","email":"","orcid":"","institution":"Pamukkale University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Elif","middleName":"","lastName":"Önder","suffix":""},{"id":693531619,"identity":"82cce4a8-5bbd-4a7b-a57f-70badef7a82c","order_by":3,"name":"İsmail Kubilay Baykan","email":"","orcid":"","institution":"Pamukkale University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"İsmail","middleName":"Kubilay","lastName":"Baykan","suffix":""},{"id":693531621,"identity":"aaa474fe-8df2-4dce-959f-e47314c47ae8","order_by":4,"name":"Hande Şenol","email":"","orcid":"","institution":"Pamukkale University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hande","middleName":"","lastName":"Şenol","suffix":""},{"id":693531623,"identity":"3a13d202-bf3a-4a1a-a23a-d9ec7ee51604","order_by":5,"name":"Abdullah Coğuplugil","email":"","orcid":"","institution":"Pamukkale University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Abdullah","middleName":"","lastName":"Coğuplugil","suffix":""},{"id":693531627,"identity":"5243755e-b49d-4966-b0c9-9aaf60ff314a","order_by":6,"name":"Gülçin Abban Mete","email":"","orcid":"","institution":"Pamukkale University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gülçin","middleName":"Abban","lastName":"Mete","suffix":""}],"badges":[],"createdAt":"2026-08-01 08:38:26","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-10557372/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-10557372/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":117060963,"identity":"d2595e34-525d-460a-a4c2-ff6a7619ed68","added_by":"auto","created_at":"2026-08-11 10:03:27","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":3617633,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative vaginal smears of the experimental groups. Vaginal cytology samples stained with Diff-Quick, showing characteristic cellular components used for estrous cycle determination. Bar — 200µm, 50µm, X100, X400.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-10557372/v1/ba71d76a627abcbd7a3323f0.jpeg"},{"id":117062238,"identity":"b5ded88c-a159-449d-a876-93bc8e4a9597","added_by":"auto","created_at":"2026-08-11 10:11:55","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1066940,"visible":true,"origin":"","legend":"\u003cp\u003eEstablishment of the experimental endometriosis rat model.\u003cbr\u003e\nExperimental endometriosis was surgically induced in Sprague Dawley adult female rats (n = 32). arrow: Endometriosis lesion. Dienogest: Deinogest + endometriosis group. Metformin: Metformin + endometriosis group.\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-10557372/v1/83aecced3144a60d030307bf.jpeg"},{"id":117061073,"identity":"f396dae0-e006-493a-8c20-e65d4a28033b","added_by":"auto","created_at":"2026-08-11 10:04:19","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2252996,"visible":true,"origin":"","legend":"\u003cp\u003eHistological evaluation of ectopic endometrial implants and uterine tissues. Representative hematoxylin and eosin (H\u0026amp;E)-stained sections of ectopic endometrial tissue and uterine tissue obtained from the experimental groups following endometriosis induction. Ectopic lesions exhibited surface epithelium (SE), vascular structures (double arrows), cellular clusters (arrows), collagen fiber bundles (asterisks), and gland-like structures (hollow arrows). Arrowheads indicate characteristic histological features of the lesions. Scale bars = 200 μm and 100 μm; magnifications ×100 and ×200. Dienogest: Deinogest + endometriosis group. Metformin: Metformin + endometriosis group.\u003c/p\u003e","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-10557372/v1/b07b0056cb00b61254977836.jpeg"},{"id":117060860,"identity":"a062df1b-6ce4-4744-a1e4-330da6f03746","added_by":"auto","created_at":"2026-08-11 10:03:00","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":3402245,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemical localization of PI3K, p-PI3K, AKT, p-AKT expressions in ectopic endometrium tissue of the groups. Dienogest: Deinogest + endometriosis group. Metformin: Metformin + endometriosis group. X40.\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-10557372/v1/569b4cd411e7e1b43ce94306.jpeg"},{"id":117060859,"identity":"2f3daee0-4d54-4cd6-8d55-f1099a019c24","added_by":"auto","created_at":"2026-08-11 10:02:59","extension":"jpeg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":3089737,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemical localization of PI3K, p-PI3K, AKT, p-AKT expressions in uterus tissue of the groups. Dienogest: Deinogest + endometriosis group. Metformin: Metformin + endometriosis group. X200.\u003c/p\u003e","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-10557372/v1/1d22210f4d31e974510dedbc.jpeg"},{"id":117065952,"identity":"3db8584b-84f7-46cf-8c6c-036fdf05042b","added_by":"auto","created_at":"2026-08-11 10:35:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":18328966,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-10557372/v1/69faab88-f86e-42b6-a7e1-098c14af0b76.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Comparative Effects of Metformin and Dienogest on PI3K/AKT Signaling in Experimental Endometriosis","fulltext":[{"header":"Introduction","content":"\u003cp\u003eEndometriosis is an estrogen-dependent disorder that affects approximately 6\u0026ndash;10% of women in their reproductive years (1). Characterized by pelvic pain, dysmenorrhea, dyspareunia, and infertility, its manifestations can extend to systemic inflammation and immune irregularities, increasing the risk of cardiovascular diseases, rheumatoid arthritis, and even certain cancers like ovarian and breast cancer (2). Despite its significant prevalence, the underlying mechanisms of endometriosis remain poorly understood, with its pathogenesis linked to hormonal imbalance, chronic inflammation, and metabolic alterations (1, 3).\u003c/p\u003e \u003cp\u003eEndometriosis is characterized by the ectopic growth of endometrial tissue outside the uterus, most frequently involving the ovaries, peritoneum, and pelvic cavity (3). Conventional treatment approaches include surgical excision of endometriotic lesions followed by hormonal suppression, yet these methods often yield suboptimal outcomes with risks of recurrence and significant adverse effects (4). Consequently, there is a need to explore novel therapeutic strategies that address the molecular pathophysiology of the disease.\u003c/p\u003e \u003cp\u003eOne promising area of research is the role of the PI3K/AKT signaling pathway in the progression and management of endometriosis. This pathway plays an important role in regulating cell growth, survival, metabolism, and angiogenesis, processes that are aberrantly activated in endometriotic tissues (5). Previous studies have highlighted the therapeutic potential of targeting this pathway in various diseases, including cancer and chronic inflammatory disorders (6, 7). This study focuses on assessing the effects of Metformin and Dienogest, two pharmacological agents, on the PI3K/AKT pathway in a rat model of induced endometriosis.\u003c/p\u003e \u003cp\u003eBy investigating these molecular mechanisms, this research aims to contribute to the development of targeted treatments that not only alleviate the symptoms of endometriosis but also address its underlying pathophysiological processes.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eCreation of an Animal Model of Endometriosis\u003c/h2\u003e \u003cp\u003e This study was approved by the Pamukkale University Animal Experiments Ethics Committee (PAUHADYEK-2023-07). Thirty-two adult female Sprague Dawley rats were used. During the estrus phase, brief anesthesia was administered, followed by a 5-cm midline abdominal incision. One uterine horn was ligated and excised at both the uterotubal junction and cervical end. The excised horn was immersed in sterile saline, the endometrium was exposed by dividing along the antimesenteric axis, and 5 \u0026times; 5 mm sections were prepared. These explants were sutured to the omentum with 7\u0026thinsp;\u0026minus;\u0026thinsp;0 Vicryl, ensuring the endometrial surface faced the peritoneum or mesentery. Each segment was fixed at four corners, and the abdominal layers were anatomically closed with 4\u0026thinsp;\u0026minus;\u0026thinsp;0 Vicryl. Animals were allowed to recover from anesthesia (8). The rats with experimental endometriosis were randomly divided into 3 groups 21 days after the surgical procedure. The animals were treated daily as follows:\u003c/p\u003e \u003cp\u003eControl Group (n: 8): Group that received nothing.\u003c/p\u003e \u003cp\u003eEndometriosis Group (n: 8): Group in which experimental endometriosis was induced.\u003c/p\u003e \u003cp\u003eEndometriosis+Dionegest Group (n:8): Group in which experimental endometriosis was induced and Dienogest was administered orally at 0.3 mg/kg/day for 28 days.\u003c/p\u003e \u003cp\u003eEndometriosis+Metformin Group(n: 8): Group in which experimental endometriosis was created, and Metformin was given orally at 100 mg/kg/day. All doses were administered orally from day 21 to day 49. After day 49, vaginal smears were taken, and rats were sacrificed after the diestrous period was detected (8). Subsequent to a re-evaluation of the endometrial lesion area, the left uterine horn was excised. The results regarding the endometrial lesion area were measured by the surface area (length x width x height in millimeters) in each group. After the tissues were fixed in formaldehyde, sections were taken after routine tissue follow-up, hematoxylin eosin was stained and evaluated. PI3K, p-PI3K, AKT, and p-AKT reactions in the tissues were evaluated immunohistochemically. Some of the uterus and endometriosis tissues were separated and stored in Trizol at -80\u0026deg;C. Following the isolation of total RNA from the tissues, alterations in mRNA expression were analysed using reverse transcription-polymerase chain reaction (RT-PCR).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eVaginal Smear Application\u003c/h3\u003e\n\u003cp\u003e28 days after the experimental procedure was established, vaginal smears were performed for 32 rats every day at 10:00, and estrus cycles were observed. Rats with dense, fragmented leukocyte dominance as a result of the smear were evaluated as being in the diestrus phase and were sacrificed (8)(Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\n\u003ch3\u003eElisa Method\u003c/h3\u003e\n\u003cp\u003eThe collection of blood samples was performed by intracardiac puncture in rats under anaesthesia, and the samples were subsequently labelled for identification. The samples were then subjected to an incubation period at ambient temperature for a duration of one hour. This was followed by a centrifugal process at 400 g for a period of 15 minutes, with the objective of effecting a separation of the upper layer from the sample. The sera obtained were then aliquoted into appropriately labelled tubes and stored within a temperature of -80\u0026deg;C, pending further analysis. On the day of the study, the samples were allowed to reach room temperature prior to testing.\u003c/p\u003e \u003cp\u003eThe quantification of progesterone, estradiol (E2) and VEGF was conducted by means of enzyme-linked immunosorbent assay (ELISA) kits (BT LAB) in accordance with the manufacturer's protocols. Absorbance readings at 450 nanometers were performed using a Biotek Elx800 Microplate Reader (BioTek Instruments Inc., USA), and serum concentrations were calculated using the Gen5 data analysis software.\u003c/p\u003e\n\u003ch3\u003eReal-Time Polymerase Chain Reaction (RT-PCR)\u003c/h3\u003e\n\u003cp\u003ePCR was performed with the Thermo Scientific PikoReal 96 device, which can read 96-well microplates. PI3K, p-PI3K and AKT expressions were determined using RT-PCR. Total RNA was isolated from the relevant tissue using TRI-Reagent, and the quantity and quality of the obtained RNAs were subsequently determined. Afterwards, cDNA synthesis was performed from total RNAs, and expression analysis was performed with reference-housekeeping B-actin RT-PCR (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e\u003cimg src=\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAr8AAAEZCAYAAACetE2AAAAQAElEQVR4AezdCbxu1fgH8LUyZSghZIjMCckQmW/GDFGGzNxCpcgliUxlnpUhswZEISEZ/uJmLJkyhUSZZxmjQf/73bfnWHff9z3n3DO85z3nPvdz19l7r3n91lrP+q1nPXu/G1yU/xKBRCARSAQSgUQgEUgEEoH1BIENSv5LBBKBRGC9RSAbnggkAolAIrC+IZDkd33r8WxvIpAIJAKJQCKQCCQCEFhPXZLf9bTjs9mJQCKQCCQCiUAikAisjwgk+V0fez3bnAisjUD6JAKJQCKQCCQC6wUCSX7Xi27ORiYCiUAikAgkAonAcAQyZH1CIMnv+tTb2dZEIBFIBBKBRCARSATWcwSS/K7nAyCbvzYC6ZMIJAKJQCKQCCQCSxeBJL9Lt2+zZYlAIpAIJAKJwLoikPETgSWPQJLfJd/F2cBEIBFIBBKBRCARSAQSgUAgyW8gkde1EUifRCARSAQSgUQgEUgElhgCSX6XWIdmcxKBRCARSATmBoHMJRFIBJYmAkl+l2a/ZqsSgUQgEUgEEoFEIBFIBAYgkOR3AChre6VPIpAIJAKJQCKQCCQCicBSQCDJ71LoxWxDIpAIJALziUDmnQgkAonAEkIgye8S6sxsSiKQCCQCiUAikAgkAonA5AisK/mdPLcMTQQSgUQgEUgEEoFEIBFIBMYYgSS/Y9w5WbVEIBEYNwSyPolAIpAIJAKLHYEkv4u9B7P+iUAikAgkAolAIpAIjAKBJVJGkt8l0pHZjEQgEUgEEoFEIBFIBBKBqRFI8js1RhkjEUgE1kYgfRKBRCARSAQSgUWJQJLfRdltWelEIBFIBBKBRCARWDgEsuTFjECS38Xce1n3RCARSAQSgUQgEUgEEoF1QiDJ7zrBlZETgbURSJ9EIBFIBBKBRCARWDwIJPldPH2VNU0EEoFEIBFIBMYNgaxPIrDoEEjyu+i6LCucCCQCiUAikAgkAolAIjBTBJL8zhS5TLc2AumTCCQCiUAikAgkAonAmCMw5+T3k5/8ZLnJTW4yLfeb3/xmUnjudre7TeTzsY99bGDcs88+eyKOcr/3ve8NjDddz3PPPbe8//3vL0996lPLOeecM91k5W1ve1u5xS1usUZd1OemN71p2WGHHbrw//znP+uU381vfvPyoQ99aNppMmIiAIFPfepTZbvttuvG4m1ve9spx9Cf/vSn8rrXva4Yq8bwC1/4wvLnP/9ZVrN2//jHP8oLXvCCri7mQ9/d6la3Krvuums5/vjjy0UXXdSV99a3vnVo/H76H/3oR12a+HPGGWeUe9/73gPTb7vttmX//fcvP/7xjyP6Gtdf/OIX5cADDyx3uctduvT3vOc9y8EHH1zOOuuscuihh3b165ffPt/rXveaNP81CsuHJYlANioRSAQWBwJzTn6Rx2222aZ89KMfLaeffnrZeeedu8XGgvPZz362fP/73y+vec1rygYbbFAuuOCCSVFCeKXj/va3vw2Mu/nmm5e99957oox1IZhthkj0q1/96nLjG9+4POpRjypf+MIXyoUXXthGmfR+jz32KF/84he7OOp72ctetpx88snlfe97X/nXv/5VnvzkJ5fb3OY25be//W0XZ7I/v//97zv8fvCDH5RXvvKVk0XNsERgLQRstvbaa6/y05/+tPzkJz8pw+aOhH//+9/LS17yko60GWtXvOIVOyJ82mmnCZ61u8IVrlBe9KIXlTe/+c1dfcyNa13rWuXTn/50Nz922mmnctRRR5Udd9xxgmCq733ve98uzqmnnlpsgqXjPvjBDxZ1e9nLXlZ+/etfl/58v9GNblTIjTve8Y4FEf7DH/5QDjjggG4+m3/m+F3veteObEfj5PGOd7yjXPe61y3vfve7u/n/zW9+s5BXV7rSlYq62Awj59/61re6Z7iqzwMf+MCCgNsww/tVr3pV2WWXXcp3vvOdyD6viUAikAgkAmOGwJyTX5oQ2pMtt9yyI7j99l7ykpfsFrrnPe95ZaONNuoHr/FsIV7DY8ADEn25y11uQMi6ef3whz8s//3vf8vVrna1dUvYxN5kk02KxbCU/3lus2ojsNtuu5XLX/7yHfE/4ogj/hc45E4dkOnHPOYxxWI6JFp6JwJDETAnaq1DwyPg5z//eTn22GO7jSiiaswhekhkxJmLKy2qudrmhVga5/e5z30676c85SnFadD2229fnvGMZ5Qtttii8+//2XDDDctDHvKQ8sY3vrGYc4PCbQBq/V/7r371q3eb7mte85rld7/7XTEPzXeb9be//e3l2c9+djdHaZ1tVM1X+T7+8Y/vTm1ucIMbeCxwpUHul3vrW9+6fPjDH+7iIOdIeveQfxKBRCARSATGDoE5J783u9nNuuPTqVr66Ec/ulu4aFdohvbcc89OQ0ObMiitY1jaGVpeCxTt6KB4rR9CK+8VK1aU9773vWtpidq4FmBHove4xz1a71nfW/CvetWrFsRCZjRG3/3ud8uznvWsos377bdfoeH9+te/XvbZZ5/CbISpg6uF9qSTTpKsaLv43CGHHFI+/vGPd6YZT3va0zot1i9/+cty5JFHdnmGVqxLePEfCzIsEBtaqr/+9a9diEVantzrX//6ggzQxNnAuO8iNX9o5RwBIydf/vKXiyN22rWIgrwcfPDBnTZePyEaEebqNOClL31pV0/h2o40nLXqaFk4sr/nqrHAOQo/5phjurie1U+ccDR/b3jDGwrSpE5hpnLiiSdOpNGn8ERunv70p3cbkEjv6lj+Pe95T4e9MuDxz3/+U1DnfvWrXxXlGndMW9px94lPfKIjacgTTeawsUuL+MxnPrOrk3qIC1/lGZtf+tKXurL8MU5h6+SEBlO/8ufURd9r71ve8pa1NLrmkvye+9znFhpTmkrphrkzzzyzPP/5zy8IsDjaeeUrX7kYP9e+9rU7zaf6wu0DH/hAtzkU7y9/+Ut5xSte0bVHG4477rjyxz/+sTCXeO1rX1ta/MSfzBnj17jGNSaiGF/I5XWuc50Jv2E3y5cvL9OJF+nNQVpnzzHenEQZQ+TL/e53v8IJbx0tsk1trf8j02143Nvoxn3Mr3he767Z4EQgEUgExhiBOSe/69JWBNDC6liTtvNNb3pTeeQjH9mZMPTz+cpXvlLYB9K4IIwvfvGLJ11kv/3tb5eHPexh3QJu8UKOHL32853vZ6YTjkORA2Xd8pa3LNe//vW7ur/rXe/qCCui4jj1sMMO64gsrds3vvGNTuOE/ElHG+YeAQuCgaDADKF1nLv11lt3RBo2CNl5550naUeEkJRTTjml05hJjygzO2FXjADIF6F2XMt84+Uvf3lnrtFl0PxBnBEhR8GOlREtR+eiWPBpyuRD84ao0aIFiRb/wQ9+cFcfR8Paa9OBCCLN8mCrivSpDwxspthxe0Y2xeEQwYc+9KHdpuDud797gQ1SKEwaJFsaZfz73//ubFj1vzhBksV99iqNHyLHzhNJheURR6zWzmsPgqVc7YWb9Npjs0JrSZOov2B+9tlny3ItJxxhtIFhBkPjaBw7Ylc/mzSJkGImCI7njRv5x7hBgpkQHX744cVYsEFRF+k4BBT2+pgJgXTKETbMqbv5FuHwXLZsWfeono997GO7MbDlqlMcdrv6SaATG+PG+IWxjZr+0z7jig2xeNNxyG4QUf1mbkwn3Uzi6HfjW9qtttrKpdgo2QR40Neufee0yjivdXLyy1RK2stc5jKFrbX7dIlAIpAIJALjh8B8k9+hLaYBRaKQGFqX29/+9uUSl7hER3w/97nPrZXuAQ94QLegIASOJBEHZGqtiKs8LHLIG60iMsPGj9Zn3333La2WclXUeft//vnnd8erCA1CQMOF+CEs6o+40gqzB0Ye2Rkjr7RHNgIbb7zxGnW74Q1vWC51qUt1fhZWx8iICoKDPEiHWNOaIXu0smwZkQtEmP3xc57znIK0sJP2gpMwWumrXOUqXb5IOk0akoWURnld4MV/HJHLn4YQ2UKY9JtgWtvPf/7z3ctWCJiyaDVhgIjbjNDW3v/+9y8IK+0/Uqf9oaWES2vu0tZPGZw2I/tf+9rXOjMTuDpCR9iUv9lmmxXP4iKZNj80fspAdIwLYcYZQsxUxwaM5lLfIDvCkeKVK1eWO93pTsX40x4Eh4YVaaLVdgQv/yc96Undsbl0fYcsaletq8mTNPqg1tXPEV/9YaHvaRrlqQ+0V11sBvg78heHxvqkVScDyCatLWyF2UAg69EvkX//yi5df4e/Ohl7bFjZ5bNPf8ITnlD0l/5G8G0E4HO9612v1Lq6/jYD0iLIzAPULfIcdDUWaND1g3lqY4toa8ulL33pQUlm7Ac78sBm+IlPfGJnimX+2Hibf+aJcaEAfeQ6yMV46ofRciP+cHnQgx5UzFN9oR/6cfM5EUgEEoFEYDwQWDDya6GxACEuFkuaOoTRYmUxHQaPhXfTTTftNFIWrkHxfvaznxXaNHkh2Y6QLeTi0oy5zrdDRGiLaOh22GGH7sU3ZL9fLnKDfCJd2oPw9ONM51n6WleTEfGRfO23sUAq+MH4Ix/5SEE4EWaOfzhkwBG0t+VpX/VLhMUVmaSp9kIVDa/jfC8KIQFIvv6BtbZ6e572lLYNYbRZUc/ZavdsmGheaUe1T1napI5f/epXXYY6aZB3EWhyXREu9UbykOPdd9+908zTsuoffRllIFLMVhDaWmtnNoDAedMfCZbfTB2CpSybNBphmxXaUG3TXv1Jo6su4inHpgZZRcA8s211nY1DssMUop8PDXPfDxa0nU4XbI6MkX6c9tk4cOJjM0FzbOPHftc8aOPNxT1Z4rTCy3w2QMphZgNX+dvAuHKIv+u6OOOaqZTNnrFhU+m0wzhfnU/+TQQSgUQgERg3BBaM/NJ+0YJ6GQxBtOCGBma6IAUB6MdH/JAi/o705YussZOkmeI/3w4hfOc739nZGtOY0RwiUv1y+c3Hoh/l0CzbDHim/UL+aPNg0S932AtG0oaDI+0Wrat8mDbQ5OkLWkvEUnmIgGNk4cgBLR8zi8hnNlcEkNZaHgi9stRFm5gv8J+OQ+KHxUOwtQd5ivYYq+yZmR3Qjrsqm8YbYY0xNyzPqfzZbzPtQQ6Z9tg8Kd94Vhfpo71OQLT3zne+c/cFEZsM4XPh9KV2DcqLCUrfn6aeFrnvP+wZ8aQdddpjPNKaOiEwT4elmak/EoqMIsDGoj5std3tvU3EupZjXh900EHFaYO0yPUw8xfh6RKBRCARWK8QGNPGLhj5hQcNlmNzizttIw0c/+m6YVouCyonH8fZNFLhfMuU//riEIognY7CAwdXBH1dcWACQGPq2BsJRQZpv5ShLPn5Vqz8w0mDxEW4OLNx+pYWVB5sN6Mc13Uhv0i7PJw4qL/7cO0zcxJ5h7vDHe5QaMiZHTi50DYv1SHBkX4mVxpxGyZmC8gse27a9La9g7AVPlfYqre2B76e58PRsjJdQk5tNGwQV9unKAAAEABJREFU17VMGwDzuXVeklyX+jK/ifjs7uN+ulemJbS+O+20U5cEiTcumBx1HvknEUgEEoFEYOwQWDDyy750xYoVxSJPg8a2c7IFvF0Y3SPKSMIgRGlh5CfMkb/47pEczv26OumQdIQP2VjX9AsVn60xTa3y2au6cohf4OJ5uo4mmbbLxgXONLA0XWw+ERn5ODJn0+leGcry/WSmBfqYn7Bhrtb/mW+II41rOJseJgKemYroG/fypXl2Px3HzEE85h9x/K0s5E97YCdc+/S9e2VoDzMPdrBe+mImIZ0X5sQZ5GqdvE3SmAuINvJkw6BdNObaG6YE7GPVT3x10V7H+REeYeojDtfee57KIdjyHBSP/fwg/2n4rRUFaWXLbFwwR2IzvC5zi8kETXjrjM21CprEg+xhfiEKbfQg0gpTpi7iDHIbbrhh96ULWmQbIWNiJkR6UN7plwgkAolAIjD3CMwr+UUSLOhRbSQi7n0WykJjAUegLPS11u5XlCw2ES+uFh8LPaLCvtH3SGn9LOxtfPferqeds6h6MYldJNKGCLNDjTz717a+ykJwIs4JJ5zQ/YKbo9phx+XSI37SODb2hr/7QU57+Wu/dO7D8dMuz67axLnnJ5zj55lzL9y1fWYfjRDYLLBhRaYs0LBQP2mCcKhH1Eseg5yjeYQTSdt1112LhR9R8kIhO05mHEggkqaOzE703e1ud7vCzhum7LEd49P49cuQH1LNX12QaDavntXPFTGlsdO/7IvZoWoTsw6fdNMeZYurfe7lFc/C3fu0HNMP9fPFAmNEWYiLY3yfHNMe9rbGkHwc+2uTz2P5LJl8HvGIR3Sf3AoSxa/v1JlNtfooxxG7/DxHfRzPG2e0yo973OO6FxyNZRp6piO0jOrHHlt71ZM9ta8xeFmy1tr9mIOxx3ZZ/sYxXIaZZAQu6itPVy9P+m5trbWYL8aJOiPh5p04Mc7c65dog+dBzpzVVmHqJD/3zEe8XAkbn1izqTJGhHHKibietd91MqcuMbbUzYZVPoPSMINiYmKDSD6x39ZmeYhvk85O/jOf+cyEbJKffIX7VJ572CC9NkRklK926ANx0iUCiUAiUEpiME4IzBv5RRKQB6QiGuzlKC/xIDQ0gUEWECFaIAu4RceC4w126Ryv+8YqUomsWJy8VY1o0TTSyAUJEZ8dpEWcVtlnnOTnxTraNNpgR8vi9Z3FzZchfLdWmAVQ/dnzebZ4WrwRCosdv9b5ogEbZuSFv4VUHSI9v3Cw8VUCeWq7o212nsLlzYwAQfSM3LFX9BkpcfkhM75Hy+TAMyfcka+0ntWfZgwpZQ8LC5sNWkr5+cwV7an2IozSIGS+Bex+mENMEC7fh/VJNG/Qh6mBcpBjOB144IHdp9psfth46kcvHfmygrpySES/HGTUC1DIhI2LNsFJPBo+5Mg9EwTjAQFGNHy1ARGxOZEu2mSsqat8jAX1cW/ciKu96uSbw354xcZG2fKFlTEhHa2kr2boA2MVgYOF8aZsX9lAXtVtkPP9WE75yDo7YvGMVXMEYULykDs4eSkL0WQWJB5ba3VBEpkI+HSf+DYVCBeb4+233777VB6zFO2mifSypc2JvpdP6+CkrPDzspZxJb7+88UCY8w98m480Qojw+YTXKQ1how7/e65dXBi02+cRD8a2+yZpUP0tUv+0pERNLDmP/JoDrUnFr7IoU7DyDySra/0i7mqj5BSfeNZGa2zuXn4wx9e9K35ax4bw+Kriy9wSOdrFLXW7pcajV1jTT7iG1/6zRdYpJMnO2b2xV4AFC9dIpAIJAKJwPggMG/k18tTyImFnbaR8wa7BcERrfCjjz66IHJ+iICGCylDWpCQ+NKAz1E5GrXgIaMWRvcIVa21yMtiJH9OWi/g0IZ5wYUG0uLnE1EW7mHQ+7YqQm1Blo8X8JAMBEQaxIXGU31olfm1zueT1M+LYNJziIVFs43nXtsttkgw7R2NF8IhDOlim4uYyEP9fUlA/WlU+cGUxgzp88whQ8xA2J56pqlEeGutBVZwo8GEhbYgfIgUW2sLuDRIBm2uegxzMKWVZG+pP/10LBItPhKGKNEq60u2kMgZba5wbUD8fJMX4YjNj7BwvhrgE2iw9r3d5cuXd4Qu6rds2bIuKtKEuCLgxhHta3xtQV9EP8IBGVIfGlsY6CdjREYIuw0Lh2DSsCKNwrRHGXBB4GjQkVztQZQQOljoPwSY9lC6QU592fMi14ifNDTONhI+K4Zws/HVXnPEvT4LLbjNiv7WXm2BXzsOEXl1FO6LIfrFWNXfCGTk09bNy3XslGHLKTPMGmh+kV2bAhtO9rn6T3obKu03fqVDmOFS65qmHeI6caCl10b4i69vtV8aYxBptxkUps4wNf+12WY5xqdwG0V21r4wIf++02fwox0Xn7OR0OfK6sf3bMypg3Yam+QNWWDzwymPjbK48tFe+XJIrjg21sLJMv0qDHZeSOSfLhFIBBKBRGB8EJg38muBpN2lJWsdkmtBtBBZ3JAwC4cjXc9IKyJIewImRFFeyIMwBAPhrXX1QiuvNn/h/KRFyjwrQ3pl8h/kaBzl3+bl3rGo+I7B5UWDGXXjH25Y+kGaZu2Bg/w5+YZ9qTpaaPmHQ1xgEs+usNIu9+HgGPeuQeJqrd13b5UpjfyQbHVHusQNN6i+4oWDI+IBF/XWlghzpY2HkfyYOURfCNPH2gFnV8/8+w7W8jd+1FU75Md5jvjKUl/5KSvahACJO8wxI4h6IT60jvLQniDFbRmD2gNrabVDPZmX1Lp6TEba/lUc5RjT+ti9OipXm7XBVbi+GlSXQe1VjnGjL+UJL2XJx7hE1mtdu26DcJKH/Djh6qZ9/GtdnYfxL291DyeuNH2nXvos4rVXdRNfXwTGwtVfGbDw3HcRLm3f6RNjp5/Gcz9u/9lYNha0V3xX7VaXiMu/75SnXHHMDXMs4uhn/ukSgURgAoG8SQQWHIF5I78L3rKswNgjEMfgjs8HHcuPfQOygolAIpAIJAKJQCKw6BBI8rvoumxpVJjZx9FHH901hn2nY3E2rJ1H/kkEEoFEIBFIBBKBRGCeEEjyO0/AZraTI8DumM0wO1B2uH6uOI6OJ0+ZoYlAIpAILA0EshWJQCKwMAgk+V0Y3Nf7Utm3+lRd64bZAK/3YCUAiUAikAgkAolAIjBnCCT5nTMoZ5NRpk0EEoFEIBFIBBKBRCARGAUCSX5HgXKWkQgkAolAIjAcgQxJBBKBRGCECCT5HSHYWVQikAgkAolAIpAIJAKJwMIiMG7kd2HRyNITgUQgEUgEEoFEIBFIBJY0Akl+l3T3ZuMSgURgcSGQtU0EEoFEIBGYbwSS/M43wpl/IpAIJAKJQCKQCCQCicDUCIwoRpLfEQGdxSQCiUAikAgkAolAIpAILDwCSX4Xvg+yBolAIrA2AumTCCQCiUAikAjMCwJJfucF1sw0EUgEEoFEIBFIBBKBmSKQ6eYTgSS/84lu5p0IJAKJQCKQCCQCiUAiMFYIJPkdq+7IyiQCayOQPolAIpAIJAKJQCIwdwgk+Z07LDOnRCARSAQSgUQgEZhbBDK3RGDOEUjyO+eQZoaJQCKQCCQCiUAikAgkAuOKQJLfce2ZrNfaCKRPIpAIJAKJQCKQCCQCs0Qgye8sAczkiUAikAgkAonAKBDIMhKBRGBuEEjyOzc4Zi6JQCKQCCQCiUAikAgkAosAgSS/i6CT1q5i+iQCiUAikAgkAolAIpAIzASBJL8zQS3TJAKJQCKQCCwcAllyIpAIJAKzQCDJ7yzAy6SJQCKQCCQCiUAikAgkAosLgcVOfhcX2lnbRCARSAQSgUQgEUgEEoEFRWBk5Pc///lP+eQnP1k+8pGPdO4Xv/hF+ctf/lJ++9vfzhsAn/rUp7qyjj/++PKb3/xmRuV8+tOf7vJQd22YUSYLkOhHP/pR+ehHP9rVHebqf95555V//etf5ac//elIa/THP/6xfOYzn5moyxe+8IXyj3/8o6vDhRdeWL7zne9MhP3f//1f+ec//9mF5Z+li4AxeNxxx030uzEa7ktf+lI3TqfT+j/96U9dHuTJdOIvvTilnH/++WXlypUdDjD84Q9/WP72t7+VX/7yl0uxudNukzHxzW9+cyI+bFpHPsLpr3/9ayF3TjzxxIm4s7n59a9/XU444YSuPz7/+c+Xv//97xPZWe/U4ayzzipk3ymnnFLOPffcifC8SQQSgdEgMBLya/Ifeuih5TnPeU75+c9/3rm3ve1t5clPfnL5+te/Pm8t/dWvflUe/OAHl0c/+tHl1FNPnVE53/rWt7o8dt11125BmVEmI0yEOL7rXe8q6vvKV76ywxrmNgLPfOYzy7777tttQkZYpfLvf/+7WBBe9KIXdVg+8pGPLB/84AfLBRdcUP773/92m6BPfOIT3WIhHv9R1i/LGj0CZIJx+fSnP70bE295y1vK2WefXd7xjneURz3qUeUpT3lKRw6mqhkCY44b8xdddNFU0ZdcOOL0nve8p+y///4Tc/0DH/hA2WuvvcqXv/zlJdfe6TYIwbS+2BxJY50xTlp3yCGHdBuHb3zjG2W33Xbr1glkWPyZujPOOKM87WlPKw996EO7cf3whz+8vPvd7+42c8YnOacORx55ZCf7KGZe/OIXlz/84Q8zLTLTJQLrNwIzbP1IyC8C+aY3vakTyAQDt/vuu5cf//jHM6z29JI94QlPmF7ESWLtvPPOk4SOXxCcEdwrX/nK5YgjjugEMbwJ2Ac84AHlve9978grfe1rX7ssX7683PWud+3KRnAt1l/96lfLpS51qXK3u92tPOYxjyn3u9/9yuMf//hyxStesYuXf5YuAre85S3LPvvsUzbbbLOukbe97W275+c973ml1loOO+ywcvjhh3dhk/255jWv2QVHPt3DevSHBhGJ23HHHSfm+p577lmC9K1HUEw01YkCGWgDda973avzt7lCPL/97W+Xb1/sbBqucpWrdOH+bLTRRuVyl7uc27Wc8bhixYq1/FsPGnhkFukm2/bee++O1BrHNL7IL22vNOQz2aev9CHFAP90iUAiMBoERkJ+7WodfdtVO3rXtOtc5zrlpS99aUd++J1zzjnlnIudo3kaQcdR/Dw7JnfPj7aD5sizPAmVCPcsrTJaR5sorTQ0kW0YgRT5iaM+bfhiuXdc/OxnP7s7Ztt1lab6Rje60UTVN95443Lve9+7CJ/wXHUDO/jChVvl1f2nQfYMD+GBD/8uwsV/YB3Yu17sPfBy6UtfutM6X+Yyl+kWBWQ48rvkJS+5Vhph6sC5V1eR9I968Vcv/eme089tuDBpwulr8Tjxwj+v44HA1a9+9cKpDY2c/tRXnLmt/1z1u/7bbrvtupOD5cuXd6SZv7icsenKGcPyjHBXz62Tr7jGsbTCmDrx48idqE/kJ85COnVm3qDO6qouNgLmOXLlOZy42iFutC/CtE0YZzcBV1IAABAASURBVM54lp944vPnYG4uuufgEXm4RhnSe+6nl6d0HLIoTuv4h2v99Xv4R95teNz/5Cc/Ka997WuLzcAOO+zQefNDRu9///uXe9zjHoUy5sY3vnG51rWu1YXf8IY3LJtsskl54AMfWAbJIZHkRZlgY26TzqyMDNI+4Zy0Ti3ufve7F5s7J46XuMQlyq1vfetypStdqWywwQblJje5SbnqVa9a7nCHO0jS1YF2+JhjjlkQxURXifyTCKyHCIyE/BIyNDT77bdfWbFiRbEzZ4913/vet9znPvcpbEC33nrrTkCI99a3vrWzBX7Sk55UbnrTm3bHRgcddFCnJRJuR29XfY1rXKNsueWWheAg7G9wgxsUgoydVduXhCybrj322KMTPo647MTFIYzf//73l3ve856d9lF9nv/853flC19MjnZCfbfYYosCG/d9R+N+vetdb8L7a1/7WiF8CX7+n/vc57qw173udeVqV7tahzkiTZvhGYG2mekirfrzsY99rOhH6WlwTzvttFW+w/8j5MwxLAZiwf1nP/uZ2zWcellIHvvYxxZawbvc5S7lu9/9bhcHyUd6aE9ucYtblJe//OXdYuKZhvsNb3hDp0XedNNNi2P1ICquwixMyn3Ws55Vfv/733d55p/xQIAtMDK34YYbFiQCcbnjHe/YyYZb3epWnR27cYZQIDSuxtJLXvKSYnOkT5008GNatcsuu5TrX//6xQmOo3Da5pvf/OYdIWF2Ea1mIkDWOB2RJuw/2fxvs802XfnPeMYzipMVBMacUF6kX6irMU/mmVPkG9mKmMHI8XrUy5xRb+0z31euXBlB5fTTTy9MkcxtaWD0iEc8orzxjW8slBZOachw84n29MADD+xIm+cvfvGLE/l8+MMfLvqIPHCCo++khxt5JP4LX/jCgkhutkrjT8sfiRHuD33oQ538Fq686AMkk2yjMNGP1oWzzz47kk5c9QcbWm0Ncok0M42JvqYR13fMEtRPYvne/va37+rleZBTf++oOK005qw3W6ySs8wXPvvZz3bvMNRau42b90u8Y2HcSkc2Is3yNRaf+MQnFv3mmbvNbW5TaKBhM2hTJk66RGBqBDLGuiAwEvKL2D73uc8ttI+On0KAWUhUFhGhBb7CFa5QuG233bYguUgqwYvsEnjS22lb2F72spd1AgR5YeMmHmFDuCFk8g1HKBJCdu7yJKQd/yPFbIHZIiNl8kGyDz744GLhRIwjj8VwtYlQT/jAyn3f0TrQXPD/85//3Jmi/O53vyv6xaLDVISAR5L1Bexiobr85S9fvExD2EvvRTWbEIIctrQ+Njf6QPggR+v7uMc9rrP3ppnyQoq+bwm1e4sLDc3rX//6zn7OPY2OPJGiZcuWdZo+9aPRUV/5aQfyc8ABBxSaZi/anXXWWZJ19qT8jYHXvOY13bMFvgvMPwuKgDGFJL3gBS/oZIB5iIjZ3NKgqZxxZczS6tGy3fnOd16LsJAVZIf4SDQbYvLHOEB+9T/tH+Lz8Y9/XLSO/Nncme/IDOKlLmQNEkk+ifiVr3ylM8khv2hA+S20Q9y0ydUcVDfkzryJun3/+98v5of2aJ9NrE2hcPLTZh8ZJhPf+c53dnM8ZKi4iLCr+JtsslpDGs/8OAoHhHKrrbYqkQeZLR7CLZ25qp7kLRmljoii9IivTYe40iOW+lKYfqM4kb93R8gg8llY6xBdSg6mC+S5MH1qDNkc2dBHX3ohjcmBOJx7BNj9ZM6phLzUSX5klXaoq7EpLc349773PbfdS4fiB1knf8k/+XQRVv2xKbDZYw7GNnmVV/5PBBKBeUZgJOSXMCJALR6EmDbRqBCCyIpngs+iRIAQ0LR0J598cie0hYdDnmgXEBzHSByNzeabb96RawKetkH6SIME0d5IS+tn4TrppJO6t2wJS4skYYs8XW+VVlQ44djmEXmN8xX5VD+kPo4jHVnS1MCsdUiB/kA+LfYWKRjCv9XmwITGyGIlb9jQgLg/9thjuxfZ9MV1r3vdjhggzj/4wQ8ED3XyoqFDauRn02EcqLdE+pRzOqA/9R9/LzC6tg7xvt3tbtfZ6tVau+NLz8h/rbVY/CxQ0rz61a8uCLzF0OJTa+3eyqZZEr5U3GJsB4JKW2gc0JrZRBkDbVvYZBoziBvbyn54G9f98uXLi2NnxMLzDquOwfV7jCdjlT9NoQ0SjTDibNOHvLANFR7OPEAszScnCLXWCFqwK5mGMJJjTkBUhEacVjWebYrNJe2jBNA+G1ftQyThTSPJFMD8QNDksy6OzK61FnNLGTCGq01G5KO/mDrFs6sNjSviXmvtlB7kOXLJkWnqz+TB+qAvzW1kWbrWkRW03pe97GW7fhcmvn43njj1pGGFgfqdeeaZonWnS/LtHqb5J5QoThFtOuQpKU28sqxfiC1Ca7wKUx8YWRM9c/xgRmZLwy9dIpAIzC8CIyG/hAwN681udrPyqle9qjjm9iKCHTJtHo0AIU7r456woxUmrFshMV0oaAAIwsniI0XKCuFnkaz1f4uZY7IgY5PlM05hji3VB4G1aLgnkC3WnDeRLUaO3Whl9IM4BDOTkbe//e1FPwRZFDaZC5JLU4PQytuiZ/MxWTphjgEdZSIUxoHFGxEW5gjQgoyU0koZM/xn4vSx8WCBgglsEBjHorSHtdZiYZ1J3plm7hCw+bIxtUk1Fo2jfu78EBtjTZxa/zdf+3Gn8xzjzdhw3GzTt9mq43jj0vhkgtHmY1x6FsdG0f1CO+TVRh1xdVpCk8tciBkBAmz8a59NMO0ukuVFr2gfOWeTb5M7m7aQLcqyKUCkyQJkMjS7k+VtM82JIw+yWHuQRHUzZ4WRWzbwNuzaya/v+jIb4ZafeLXWzhyMLHRaxK8fn5/wTVZpuMMxu0KqySNmXdtvv32naEFqnSiwTX/Qgx7UnTRJz8HTJpzsqrVO+flGY1vb9Zv06WaFQCZOBKZEYCTkl4aFLWkIGlpWpNeu2KS38KgpWyrHjOLTBLjnP5cuhItjMWXTMgzK3+JKgA0KG1c/Jgi0uDTZ7Pim2gAgEdrCXIRZARJoMWRny38qF2SA1sOxnvS0SY6jp0ob4RZuBDieXS3UjhJpoByjOoLmPxtHu0Kr5couWF052huEYDZ5Z9rFjYBxQRYg4IiicYG8ITnj3jLk0mcMbSDV1Zw56KCDinmjXdriSpYZ59E+ZJV8jbCQi/KYiQtZwr7V5lm5tMs2NFPlJ60NBfJH9rcvtNmgmrPyoE2VL4d48mtdrbVE3NbfJp3db+uH/FLGINitv3vkVxnhlEUuiktrzOyKvIOlkwRpOOsYRYIwz7XW4iSRTGbKx2+Y0+Zaa6d8GBYn/ROBRGDuENhg7rKaPCc2WrQCEYswQYZpIGkIwp+2graScHLMGf6zuRLsjteQQVpldmfsBmk5veCG6Dr2VB+CkgC1k1eP2ZQ76rTbbbddYctK8wJv2tPJ6sD+jXDWZsdtNKS0X8O0Kv28vGDDT1p2he4Jf4Lcfd+FBqf1Zy7Bxlqdwx9xt/mhmVu2bNnMP9sUGa660uIg9epGA8a+Tn28VGNcrIqS/9dTBGz+jENkxlE7GJhlkRvux90xP2LSE/XUDmQYWUQsbUZpqhHlIIEIs9MvBJC2nZbbSZDTkb55kTzIxMifFtTciWdX5iiuNKPIonsmBmSq+6mc9MivE0IyWvxXvOIV3RcSnNJYC5g62JSYu1GGeOFqrd0L0trOhIC/+c6uf6eddipM7mBDm6ytTL/EmY6DAZnqhUKnUZ776ZjKsFOnoWZmQq44WeRHsdOPH8/q470LGmpa5vDPayKQCMwfAiMhvzQqXnhDMgkBzgsYbD2DQEUTvfnqWJptGHLKH3H1NrOjMVpBX2dwtEdgEHLeQKb1RGTEdxSF4Lhnw4d0v+997yvypoFGtpAqR2IEv+9kIoAWQQLSd3HVj7bEizDyUQffonU/zu4hD3lIsbBpi7rDmqM9ZWrCBIV9tTbQtjjitbBoN8FrU+ILChYeCyHBfPjhh3dfhKDZiGcaXtj50gZNB2KpHFoOmwv5h0NmH/awhxWLNPs7R4ERRuCz5WOf6Eibv4WaBsu9hYZ23tjxBQjxvIyjzy0uFm1jSB2RZfVnPuMlOuSFFsoxL0LvxaCnPvWpxThRJjt0i2K7sCsz3fwjYO4bbwiD0vwQgPlJI+k5nHH25je/uXtEysiG7mHVH/Pcj6Wsuu1esrLZo/U0BvjpWz9+wQbWMxlkrgfBRaakMW6dNJAHCKNn5MZYdjKB8Egvvnjux8UxXzCmaRbVmyOnaCi9BKyecCbzzBeyTRwymWy06WQuQQFBEYC4IcXShWOCgMDZHJOLMKEY8Gwum4OUBcgkUkk2K4NMkNdRRx3VbWDFp+xw6keekt36ixzXV/pFH4sjvXqTTcuXLy/mOCLpZV19RPZE/eJKlmiDeU+e86fRV3cySbm0tuLc6U536r74Ic50nLrQak8W1wmWl+a8vEnWukfEySPjaVhaZmJwsgGz7g2LNxP/TJMIJAKDEdhgsPfc+nohg/Ai4NhrcYQcIcSm146fkCC0LEA0kgRHratt+hyvW8RoEQhM2kUCFdkh0GkbLJB29fKilVCmVihH2QSSo30aXsf0QXgcq9Ey0zjIx+KL+FlUpEeW5als2iB+4+wQdiYdbKaRAFhzFg6Li2PIIJba4ejT4kWbAh9fe4CJRR7WFiykc+XKld0vssVzLD5ILXzkoZxlqzS18m0d8oqk2LwgO152a8PVGSG3aPJHypUnX1ogix2NDzLuJUmLkLYJZ4dHI4PgGAvGkLfWbQDU1RhB8C32tL82Qgg8Qo48eWFOmelGi4CNB+2dzax+NHeNP2O3rYlxZmyKo2/1ZYSzcbe5ESY9YmQzJh4/pxFIoDHiGTGzmUI2PEvrs1wIEhlgvCBixrG8lOMTYG18L9vxHxeHJNrUqb96c2SclweZNKin9jnp0mY4iUP2CjPXkUNykskB/BBiYeGkt5mUnny0iRXfs82D+DauiDM5CWdleBEWwbbRMQ/NT/PUnI5ncoHpgPTWB31gbktvHSAbyGKabemjjkwQon5xrbUWn0SkAPDpOvERYkTUBt0zUux0y+Y30s3VlSyRN1zYYltTfDbO+jWsDHGdnDGxsJGIdWlY/PRPBBKBuUFgJOR3qqo6IkJWLSwWJEKqJWhTpc/wqRDI8EQgEUgElj4CiPSee+5ZkGynQePeYpt6mwNKB9rzca9v1i8RWCoIjAX5pe21Q6fR9QIS29WlAnC2IxFIBBKBxYAALaQvt3hRTX3JY5pM94vJ0Rg74fE5S8Syq/sY/qH9dmrl9I1Z2hhWMauUCCxZBMaC/HoL2We2vIzh+Nux95JFPBuWCCQCicAYIlBrLUwGjj766OLzXUyf2LGOYVWnrBLzK/X3styUkRcoApOTvfbaq/uhoclsgheoellsIrCkERgL8kvry26M/RoYLZp0AAAQAElEQVSj/3lEPLNOBBKBRCARGIIAW1pymPNNXbbyQ6KOvTcb3HFeT5x4WvfYNY89mFnBRGCJITAW5HeJYZrNSQQSgURgTBHIaiUCiUAikAgk+c0xkAgkAolAIpAIJAKJQCKw9BG4uIVJfi8GIi+JQCKQCCQCiUAikAgkAksfgSS/S7+Ps4WJQCKwNgLpkwgkAolAIrCeIpDkdz3t+Gx2IpAIJAKJQCKQCKyvCKzf7U7yu373f7Y+EUgEEoFEIBFIBBKB9QqBJL/rVXdnYxOBtRFIn0QgEUgEEoFEYH1CIMnv+tTb2dZEIBFIBBKBRCARaBHI+/UQgTknv8cff3xJlxjkGMgxkGMgx0COgRwDOQZyDMzHGDjxxBNnRdnnnPyefvrpJV1isCjHQI7dnLs5BnIM5BjIMZBjYOzHwJlnnjle5He//fYr6RKDHAM5BnIM5BjIMbC4xkD2V/bXYhkDu++++3iR31nVJhMnAolAIpAIJAKJQCKQCCQC84jAnJs9zGNdM+uRIZAFJQKJQCKQCCQCiUAisDQRSPK7NPs1W5UIJAKJQCIwUwQyXSKQCCxpBJL8LunuzcYlAolAIpAIJAKJQCKQCLQIJPlt0Vj7Pn0SgUQgEUgEEoFEIBFIBJYQAkl+l1BnZlMSgUQgEZhbBDK3RCARSASWHgJJfpden2aLEoFEIBFIBBKBRCARSASGIDBt8jskfXonAolAIpAIJAKJQCKQCCQCiwaBJL+LpquyoolAIrCACGTRiUAikAgkAksEgSS/S6QjsxmJQCKQCCQCiUAikAjMDwJLK9ckv0urP7M1iUAikAgkAolAIpAIJAKTIJDkdxJwlmrQP//5z/LpT3+6POpRjyo/+tGPRtpM5e6///7lO9/5zpyU+/nPf7487WlPK7/4xS8G5ve9732vvPCFLyyuAyP0PD/1qU8V9Yv4f//738tBBx1U3vOe95QLL7ywF3tuH88888zyyEc+snzzm9+cUcZ/+9vfygc+8IFy4IEHll/96lczymM6idanOHe4wx0Kd8wxx6xPzc62JgKJQCKwpBFI8ruku3dw4/7973+Xk08+uRx77LEFER4ca+59//KXv5Sf/vSn5eijjy5I5UxKuOiii8p///vfiaTnn39+QRr/85//TPjFzdlnn10OO+ywjrhOpzz1kxei849//KPLRnm/+93vCtd5zOOfS1ziEt1mJMpWlPLb9vIb5v785z+Xr371q2XlypVlEB7SyfuQQw4p17rWtcqmm25aDj744MJP2EK4H/7wh+XmN7950fZaa7nsZS874TbccMNyy1vespy9qh8Xom7K/OAHP1iufOUrl0c84hHls5/9bNEf/MfNffe73y2XvOQlJ7C71KUuVWqtE8+Xucxlyitf+crS/3feeeeVQw89dOBm1Gbv9re/fdEP+kUeG2ywQXH1zP9e97pXOfzww8vVrna1rix+6sGJw6nLl770pfLyl798Ii3/WmsR5j6cdqgjnM8999xuI7flllt2eV/72tcuL37xi8txxx1Xfv/734s24dTV2H/4wx9eNttssy7+Xe5yl07GffGLXyx//OMfJ+K2N1/72tfKu9/97kKOtP7t/QUXXFBOOOGEsssuu3RYmDuPe9zjyve///1Olqnrdttt14VpB3z6ONlAkbVw2XDVuBZP2437eL7c5S5Xnv/855d3vvOd3dwUR5g0nGdOuq9//etdFZX917/+tbz+9a8v+kq4+j3lKU/p6kc50EW8+A9Zos177rlnV9+rXOUq5UEPelDRP6997WvXGN9kyIc//OGij694xSuWK1zhCt08EPejH/3oxTmueVEf4ynqt2bo6idx/vCHP3R1vulNb9r11a1udatCLunDU089dXXE3t//+7//K+9973t7voMftfMhD3lIgc3gGKVbRyh/9tprrwIH+G+77bblXe96V6dEkMegtGeccUZ5yUtesgZW/XjSfutb3yrGScyNhz70oZ0M+eQnP1msNS94wQuGzgf9Lt3pp5/ezzqf5wiBJL9zBORiysZE32abbbrFcpT1vtKVrlTudKc7zarIL3/5yx25i0zufe97l+OPP77c8IY3DK+J63Wve91CsN34xjee8JvsRv0sUm2cjTfeuCMHz3zmMzuC1obN9f0WW2zRaX3vete7TmRt4f7CF74w8TzZjfSI5GRxbHo22mijQgA/61nPKha8T3ziE5MlmdcwxEadLAy11oLwhCP49e+8VmCKzBGuV7/61eU+97lPRwIsvtPZSE2R7ZwHI3+vec1ryp/+9KcOwxUrVhQELLA86aSTBp5+nHXWWR15tSHtV6rWWhAIpyry+cxnPlNucIMbdPPPs43ibW9722Iz/f73v78r97e//W159KMfXXbffffu+ZxzzinveMc7Cn/kyXiWllO//fbbr4uHnMJZn2vLxz72sYKI2HQiYE411OMWt7hFedWrXlWER31t+g5adTqjXOPFqY38temMM84oBxxwwEDyqxxE+u1vf3uBQ+TXXtV7n332Ker2iFUboN/85jfdBl69bRy17ec//3m5xz3u0W3SlOt0CwFduXJl1zabN+MI6XMKFX3kHmmGo3SwMbZsSj/0oQ91aX/961+Xhz3sYWXvvffuns9ZhSdy+ctf/rIjX9/+9re70yKElr+85Yd0ORF74xvfONEcZbzuda/rTsq22Wabrr7wJQOceLnajErg5EgfvuENbyjaL54+QM6PPPLIwl+8vlNf8sRGa9CGwkaCPLvnPe/ZjUcnVQgqYnuTm9ykIO2nnHJKP9tujJFX+tRYWStCz0N+NizGJSLaCy7q8eY3v7k85jGP6dYOp23/+te/uhNRmxSnf4NIuDFDOaLvf/CDH/Sz7Z61B5bmjvUkxuPLXvayYv2yCaRwsEkj+/QLZ9P0ohe9qOtnbYT1T37yky7P/DP3CCT5nXtMM8d5QoAAtgjNU/brnu08p7DwWqzmspjrX//65bGPfWzZeuuti8WEdsHiPZdlrGteSBFS1U93vetdr1uoN9lkk37QSJ+32mqroh+YCT396U/vNHMWr5FWYorC9OMee+xRaK8GRUWyEIs2DBmzMCOvSD1i14bXWjtN31WvetXWe+IewUPKaAYt8hMBzY0Ffeeddy7KF+92t7tdE/q/28tf/vIdwTMuETqEVZ6Iws1udrNOQ1xrLQ984AMLgqBsqWknkbHDDz+8ILFPeMITOq2psGte85pdnk960pM67SK/1tE+0jRq9+c+97k2qLtHdOS9chWJpel78IMfXGyQtQkJjzEBR5rUq1/96l26/h/+iC6y/8QnPrFoaz+OZ4T0yU9+cmdmAy9+fadsGmibDkRYvjTByOZtbnObrv9pDeFMo33HO95xIguk/C1veUvZbbfdinqoF40yxQGi9exnP7uLi+Q+73nPKzBB8nbcccdOS6z++oL2HdnvIjd/4GA8IfAnnnhiQcib4O7WyZ860/gin052Ln3pS3d9ZuNCvtsQd5GbPzZelDY0yvqsCVrrFolFkikCbJz0bz/Sxz/+8U7LzswM5hQl4jjl8az9nvvOBsw6ZNy99KUv7Qd3pnHKNl6QXXmZmyJSwpAfxqg+sgGg8RbWd2SiMQWnflg+zw0CSX7nBsexysXOk0mDxdo9wUawOFbqV5SgoDG4+93v3h07teF2vvKgiaTp+NnPfjYR7LgIiVq2bFnZddddCyHjKIcmxaJA8BGmwuXfJwvKpT2QtyO7CLfrJhS33377bhGQl0WKRsLxHA2FRdwCaHd8xBFHFMdHhJLKWbAcy9ECEfwWBYu7sOc85zldnhZVApxmggaFRoffII2D3b2F2PG3vMWnYTv44IOLOllYLYraS/grx6L0tre9rdz5znfuNIbshS0owghNed33vvfttGrPfe5zO20vPGizaGtoIWjALDo0tAgGYUpbAae3vvWtnRZSn1rgIm/5T+WQX4uleNKFEPY8bs4iR2PmyHWh62YRMl532mmn7sjTuFvoOrXlI3qOvFu//r1j8dbPeKWlMy8cv9NituG11nKd61yn9VrrXv/YuAwj3RLoP/EGkRrh4WqtxYbH/KB5fMUrXlH6Gx8E7G53u1th0iCd+Uj2mAvIBL/WIRHmJ3xaf/fMC8gJeZlT/FpnvpFdiKQNUBvm3gkK8iTvzTffnNdQh4jB6BrXuMbQOPqA9hMpNS+HRQw8mTR84xvf6DS/g8rnh+RGPuQYAknWI73h74pU0/Ruser0ybwjZ52aDSJn6oeYSdc6sk19aIqVg/y14TSwJ510UmdyhxT226hvyUyuTWcNM98Qd+ODVpccbOO09zTLxtqBBx5YbKT6cl1+1gVji/zuzxvykUYY0W/zVX9y3gaNZh0h77+7gmgfd9xx3Ti26WvTu3eS6JQLyUaG+Q1z6mFjMiw8/WeHQJLf2eE3lqkdobA3QoARSZOWYHrGM57RabDaShMkhOkOO+xQkFVHRcIJP8dttIN2sj/+8Y8LzYIjPMIDSUOMxSf8EWPHj7SVjpqQNflaIJgM8EMe5a1+jvUQMUIGmfzKV74iqFj4PvKRjxS2usi6fJSx5ZZbFoTTkeCb3vSmjvAiyo6FLAJBcBF27SR47O5pIdRZ5tpCuKsHAUS4WJSRAm2hTRGvddpDmNJY8HesSJiq4/ve975OMyYfghbGSCxizh5QvZRp80GjYTGFIyy0wdGYfkJC1RPJ12eeg2zRGuhDmgiLowVFGkdvynE8JwwW6jcdZzywX7WoE/60L9NJN4o4MORsAGxiRlGmMmis4Ftr7Wxla137auyxBzTObcBoS6VdrO6oo44qjl4ReqTLeEI6F7I95sBpp51WzHcEqq2LcWtsuJpz7s1t2kAmEm1c9za34ogrjXnPn7NZtkFlI0yLedZZZ3V2nsLCIU7yR3xpfMO/vSJONJet36juyR8y1uaj1jpRrLay5ebUz5WywoYa+RZ/InJzs+HFtsjknX6ggGiCu1t5wZT22lVZXcCqP/Jns/2ABzygWE8QXXmtCur+6w/zGoFFzDvP3h9YqkfrbS2y9khHIUB+6r82TtwjxZQIlCo2NTZvnikkIo46Sk/+WZ/C39UY0S5E3LhRZ/4cGYv8WidsKshpG0abSOGcfJna2IjZUPDrO9iRNX3/fB4tAkl+R4v3SErbbLPNiglqd++Y0PGMFxSQPALf4h0VYctI+4EE0Uwge14mQdaQYUdxBCbSRSDwJyBMbESPoNhrr70KbQJiyaaXUEB4pXccSRgii+Iq18RHNvlzdsAEizALnuNRgocThiwL6ztHVQh0648oLluljbagywsxjIXLFRkVH2kmtGir4WQBVC9hrbP7ly78LBw0Hwi/vKW7//3vX2AhP5oAApJwVH9CmAYFaXJcJh/x9IE8EHxaAHlaZIUPcxYOpJfWRZ222267TgvpONPCMyxd3x+ppEWxyOgjWpVWyPfjj/KZdp/bd999i4VwVGXTENIedYadcQAAEABJREFUmg+TOf2vTsado133i9HZbLEHNe8RJJtZRMYxv/m9UG2yQSQnXPt1QFrIGfOa2YMNrHFr4zuozk6EaC6Nc+SYRk6e4pqPNt3Iz/3ud79yoxvdqJBxyJM4XHvvedwcJQRZot/ausHJqRwZwWTFPTvgNs5k9+QtjPp9AOenPvWpnVYTpk6vaInlJa7xFBrdFStWdJtIG0T9KY66kpPup+u00eZGWeS0OUohYOM2KA+bWMTa5gnJfPzjH18oeNr2awfM1LnNwzNFj7WTPKB1ZV8ecWzK4EJOG1fGn/Y7HYw4xmO0N/zyOp4IbDCe1cpazRUCjtvk5YgH0TQ5ET5+HCHmSlAit+4d3dBwEiKeOSTQpLeTR9YIOaQLeWLEjxQEeSRwEfBaa3dsadcuT4JDXkgNQebe1VGQe45wo4kkNGkOghQLm46jbVUX+YqvfYSle45QVIYjT0KQRpewolUQPl0n31pXa1sIWc/S0uzCgZ9nBJiGAcEgJLdYdayI3COfFmfCnZZX3KkcjTPs9FXEtdlAZGxYwm+qK0JOe+JlExsF2maaHlouR6PhmFRMlddch9PycLT2+mmu8x+Wn35hYqLsYc5CaCNoLljonU4My2/c/ZkKMBlgJ6u/mT8gJwgiLehC1d/4NnfUoU+UyAVjEv7mkDFi3iGw4vfrjMyaY/wdc3sByT0iZDPuZSNttwG1YWfGREsoDkeOuSKDiJH7oW4BAmye1dvcbYuHk00MeU3eUX7ATByaW3LP/TBHm0xu0Ra3cWDN9If2lT+iSP64t+EnRyhZYGpzQQ7aaCON4lhfrENkeqwF/Ic549GGnymFuSZfV/WnVKFlbdNa28xdm1ImM07crE3WMukirvFifdA+sj/8+VHKHHTQQQUGzPmYpQlXJhydcDrlUxfrqDGjPGWLR84jzuS653Tji0CS3/HtmzmvGZJZa51So0Yw2RkjQ20lEFoLEz+LDyHheAuJQgxaQSIORwASCFytq8ki/2FOXoSq/GlPaXiGxR3k73iLoBoUxk89aIW95GUBJCyRc/7CZ+sIQVrdVmsEM5pyZRC8hCoNtE0E+zjEYzrlwpeTf8S3SNGEEdzhN50rkmFhpMmxKMGMI7TDIdXTyWs+4lhAprspmI/y+3lagJkBWXCdfhif/TiL5dnpjjmGSCIynEVfu5jrMIlZqLYgnDRvxh7TqH49jFvzydwR5vTH5pK5FCLIL1ytqzffnmPDiMQiaeyAvTim7Rz5hdwxJUBqpKH5894AbWL48R8Xt3z58u4TjAgegt7Wi7xBNOFF7jDv0k5KCKdebdz+/bJVJ2cUBuyg+xsQ+TohlEZfuZIbtOjGD22ocjhEmRkN21hyi6yy8da3Tsikncwh6hQgtL5sq+XplJLiBYFWvza9cc1UxSmG0yPxnZJJT5lC0SG+lxX1rTWA5pZf68ho8hQG4W+dsCYyM5Mv5z0YY9WGEcEW14kQuUXbrJ380o0nAhuMslpIlQE9yBFKURf3/Tj8IrzNp/VHPIals4tsw8TlF3nO0XXssonjcBOXIDExaXSj7YOuBANbKcIM1hrlahdOmPicjcltAWJbi0DZARNw4tJCxmKBBNK0WEQIYuHK5Nxz7jn3NAZMAJhbENr81d211tp93sczJz4nzJUjfGghonzxhIcTh8Dygpt6e6O5tW2LeK7ihpvOszhMEZg30CrBTHoLjmNV5dK2KZONJQLMZCLsnaUXP67uOW3gYC0vGveIY8HXV/IXN/zdD3LGfdRLXAKalgM5UCdmI+EsIoPymA8/dZGvdrqOi1Mvn6BClGh9fFbPYmohH5c6DqpH4BjXiKM9FnKbVuYd5kE4tvK0hdpojkeauEZe8gi//lVYuH5Y+zwsL7j6agWCYvNrbkRc6ft509gz3UDSECQbQ3EGxeVnQ2+D7Dg82u3KPMwpFhlHTsqD/GGy5DQJgTNv+MvH1VwyNhA8fuGEuW/r7bnvhIvL9cPaZ+Hitn7u2bSqn1OIQfWTTjyOEoImXF0RfHI5wl21TVusF14ohA+Cyg68H1d8eYaTjgzSD7AMZ85QNDhlMJ5qrd1LwOQtjbx1om2X+yCk1mr3Nh7GQ+TpivyGrApCq0603eQpYioepywvDOtLmltlIO/M/5B2ml3rE/9oj7zi3pWM9NURpyTyDGeTEGOUhls6igjKDXnDDq785eNqk2LtNAb5hVO+cC788jq/CIyU/BqcjjINEDtIxzMWEsfUDMQdmZhojixo4wxSu3ykwc7cwDHYvZUqPWfXz8AcTMgEG0z+HBtXg1uYSe/lKos8AuiehkvYUnZso2gWvWCFOHlpzDGZHS/h7eiQtgf2JixtKEHF5lI/ETQ0JV4800d29vAiCPSTo2KC1yIU5Fb+8kPS2JP6KgMthf5Tnl2yNOK5ErgWXCYDhBo/R5HKENdiJI5+I8SYLPiCAg0GrYcFzSJJ0Dje1B5HU9IxFXAkSjhpl7pzPjlD4Lh65ggkbVVfdXdlb2bMyZ+QVEdXmkD1JKCRWYRbWpghEeroqI1WANnweSH4EPhwo3Wj2WKf5+1hhFmZxiTTElcmK9LIS3tpbxzlyQ9BhbF8jGUbGjioi/rKS/u0q3UWBgJbWkIYPuyzzYs23qjuaXDU1dEhwW/DpV9HVf5U5ehTY0Q/6HN9MlWahQy3UJuT5jw8vVRpPESdEBUmNzTY5kX4uyJ+NO7GNXkb4eQCeSEvc4hMQUqkaZ0N57HHHtu9za98pIc2ro3jXv/qZ+PTvNf/xoEwzgkT23bk3Ng0l8kN8RA3Cg/yRlwOwSIvyBOkjdyAgfVGGvOLNo8s0QZz1ZG3tOHIRoRLfWkxYVBrLTacbPvJT1o/89BYjTkofZxOwdsRvzqSHVEW/MQLh6Crn7z0jfjWpwiPq/qSEV7iZY5BNpMTEW6jYH1DDp3CIe9wgxVyZy11ukOjLY2NDeUCWUZGeVmLll9bxVdnMlZcx/oIH+LvdEqfw5IsImesydFefamu2i5tOLJRX9HGwlA4xYaxpS607770Q0aSecpQHwoMcW2+jQvlke2Rr7GrTdpnbhqbZK21zfg3DiKu9MalujL1Eke4d0msHXgCJYjxAwcYw5qsZeIkrvrCRp3kF3lrm36zJtECO00RzrxQ35trZK26GS/y19dO2vCayMfaZmNi/CkbBv0xE3HzOncIjJT82hEZgCG4vDFNCBg43mAnxAwEEwX5MsANEkLHZKRZJNiQZbs4GisDEpkw6EwkZCDgsbtEhjwbzAi1ySKOgepYSNhSdna+bE0JSosBrS6SR2D73qLFjnAxuX3yiIaW0JLOpLVwWKwIK0JAfzCf8K3CwJ89GWFKkMCS/SwyRujROCBshCphVWvtPjqvb2ktlWVcqBfBbiz4jIy8EW2fJPNGtvyUS8tB48ERagQlgk44WkwRdMTQcSgSbINlfBBIhJn6cTZWtLS0HJ45Y0j7jDVkR/0IdUdt4iKunpVNy0Qzog6wVG9tqbV23xaFLUxpCxF/mjZCTzscMVsAEWnaEraI8lEnC5jjRHWxYDs69Okf9XQUZ0GwybORQMK9zAInYfDTX7Tz8oOHdrXOvNKXNozKgJ00bZxR3msHjMx9c1o/WlBGWYfJykL2LNLmDm3kZHHHIYwGFGk1h2xYbXC1Qd0QAH3uSNipDaLHP5x5bv4Yu458jSdh5K65RDYgW2xNnWAIax3iIY3xbgwaY8ZhG8e9tPpZf5tH+t84EBbOvCajyRxk2CYROTfXkDLzJuKSOz5XiCjaZGqj9QShNW+sOciO+lkHtLHfdvPanFcnL8BFuDlJc4gcmjfqrh4wYQZDFkQ94Ctc3ck8BNGJmD6IOK7qYe6Se4glLMgKYa0jf/iTtQguPMnuNg4yqZ+ZAJCx8oUVWUN2cjbk0vCjGbX+ke/ytqHTdmY8iL54HDnopTVrALzMSXGNL9jb2GgvrGlAo2xpwxkL1gaYuqqbMLJeHchZ65L1iSzFC8h6bVIncg+++qKVZeSs+tKsUnAJVw9ru/WmxUg/GVtwxivIyQj3EiQzO2H6gKJDHY1zJB9WSKjxpw/MD8/awCG/0pH9MDHPItyaYdzBDs7GDCINd2NSu+XBWX/1DZycQJgPiLCwdPOHwLyR3+lWmcDx5qQFXxq7I9dBjjCwkzJpOMLHoEES5DMoDT9Ew0RGhJALg4//+uC8HW1yIjmEsjY7IjfhYUizQti3uNLOi2ext0FBtmCNfPJHmAkGmMKT5tGiVOtqm14LBg28TYtFkPaBUBJHmeFsYuLelaCzSMnXImZMOBlAPvSxstXVYqQthJd0nPgECuclDGTckaA2EOoWLmNFHhzhLZ/Il1+/fspAIuTPEcaErvtwxlTcuxLc8kS2YUMgO52wQCtDm/SFPhEfTjQHMIIXP/VGMMSHs2cLg2fY8pNeGwlqbRamHOnDEfT8W6f+0up3uCA5bfio730+zBiJOruO0/ykFVInY23U2MykPOPP+FLncMatvGqtxXwKfxs6/uEQyAhzRSyE2Yi28kGYzZyw1jn6FhbO+EeK2jjukaWI46r/jQNhrTPW1d34F8/m3DznN0hxoXxj2rwQX/mezX1zxBziz9EQt2XR6PMP14ZLa4OobOHmj80aedHmQb6Ym+KEIz/IgzaejTm5GnFcyZE2jntKBGHhyGEnY8JaR36Y+1E/8Z2OkuPthl8acpjctekQj8yxGbcmCG+d9sBOuLLFt/kmg2EtnPzhz5HTbXrynH+4Nhwm8ol1CB76FemWh/EV6cgsfcCf064Ic9XHMU/Vr22LTZ4+EI9TXoTXWgv5pyzyULir9jK7o6ChrLBOCDOuYiOhHsYn/3D9cP2HBAd21knY9+Wy8iMPV0Q61lrlpJsfBBac/GoWjaAdZa21EN78BjkTzq7RLtCxrZccCBI/hNBOjjat3ZYjIdoOgwxxa8OX4j187FLtTB1beR5FO5VjF09DYTdLMzCKcqdbBo2YcWLRcuSFeE43bcZLBNYRgYyeCCQCiUAiMKYILCj5dSTExsUxNaLkyIOGYTKsHDOwa3L8hjQ78mDa4Nijn45JBM0nWyVHT9L24yzF57Cj82YtUxDPo2invmRTRovBDoyt2ijKnW4Zdu20I7Qe6kdjMt20GS8RSAQSgUQgEUgEpovAeMdbUPJLO8g+yduThx56aPFy1FRwObZxPMc43nEG0stwn01NPy3tJ4Ls5QZkWXn9OEvxmd0Q8xEOIfU8inY6ClRmOEdxoyh3umU4amO754UWpwjTTZfxEoFEIBFIBBKBRGDpILCg5NfbvEgpx25nMrvdPuRssJAY9kCM4we9WOFFIi/IsQ9jzK6Mfj75nAgkAvOLQOaeCCQCiUAikAiMEwILSn7XFQhmC8wXmEhIy5jdCwbe4O0b9gvnvFzhxYdaa/FGqTds2aYKS5cIJAKJQCKQCCQCicA8IpBZjyECi4r8esvWC3F+rYW5AzzZ/HpxKerxTMIAABAASURBVN7g5Nc6Lzh589WnYtx7Sc53I70I18ZbSvdedNM+P7TgszrD2uZ7l+yuh4WnfyKwPiLgU0O+tckxm1ofMcg2JwKJQCKwlBEYKfn1UX0vp/n+H1C9iOU7jswWPHO+k+fbez4uT8vrGWH1kpvPQTFvoMn1vVLfUvSpEs9MG9j++oi1fDh2wT42bQFDmn1iBGlWpk+r+NyVeEvNIbxsfZl9+PLCsPbBsMVrWLz0nyMEMpu1EPDtZGPQXG6dOWoMG8trJZpnD99qJUt8UsmLkfFd0HkudlbZk6ktfoPu/dBLW4h3IHyL1NdwfA+WjOXH+T5qxCWH/RiKH8eQrw/y+9qOT0jaKES8/lV5sOz7D3qmxPDjEYPCWj9fbKH8UGffcfejBL5BO+hHN2zuo87aZ73xHVeKAXn6gQntGeZ879j3jcVtHTz8kETrN+je912tY77fqgw/aqF8mFmHVq5cWfgPc9pofkTeTj79iI01TRrfL/dDPt5p8S3biBdXuFhLo3wmgJQdvpMrvm/JetdGXsOcH2WI/Jy4Gmfa5Bu78BeuT3wD2ImqH6sYlhd/32n3PdzI09qsTUwYhZtv2qQcP2YR8VyVzy/GrPK9YK392iXOIAdzXyHqh6nvVH3gx018L7qfFi/xHeq+f/usvrDGQ/Sl+vq+se+7+z2DNm7ejx6BkZJf33D1QX6Tx6+fsPVlluB7edF0BNU39nwbjzCkffHtPV8Q8J086Qw692yGfQMyvonqu5Te5BeH880/3wxFjH1j0ItY/OXpu4C+ARvlLqWrz7n5Ru1UxMHCANul1PZsy+JCwFj1nW9z0sf6zWkywcLse7VOe3zfe5St8n1Si57yEQ1fTLFIj7IO61oWkoSo+VYqDL3si6S653w/1Vd1Il8y1IuwTMH8GIOP8QtD+BBFSgbPfmmQwsIPBNRaC2x8RxseNtdIkHh9h9z5Piry1Q8b9Hz44YcX30mVblA4P1/t8aMu6uwbxTYnSDrSpN7icJQpnpVfay3qq33IOqWHH4TwaU3ERPuMNxj5FS7j0D2nLoNIrvKVOYhwK5/zpRvfsVVn3zqWnxduEVB1QO5sKKxNwjg/3mGz4F4/yh+plB/S5/uvNorM/XzFyNd8vMviC0nKES+cH4/yeU+nf96Pkad10o83wEX5Tgi9cO7Hn4T7lUcbC58P9Wy9hgkSL1/18Q1c67LvInvfxrgyR3zPFpm0odBOYepnnFmH5KfOiKqvMMnPmPVNdwRW2eIYi3DxXXl9IV44n+30tR4vt+MD+p/yCybiR7z2aqOiTcZ06+/eZkJ9fcdafWFuvBob6uIHKZBUGInfOnj77rSNYuvf3sPJj2P4sSecRn195x1ew+rbps/7+UVgpOTXpDLAWueLAO0np5BfGt42jnsf90aS3bcufjwATCZ5G+beZDIZB4WZ3NItNce8gw20L2NM1jYCzHeTJ4uTYYnAfCJgTtugWmhqrcWcZcaEbN361rcufibXAj6fdRiUt0UKWfRjBxZOxMTiPijuOPhZsGmrLdow9B6EernnnI6Rhfxo1P34CgWAdnlvwhd3dtlll2JB94tWiBHNFWLiCym0Vn6MwgKuv5Ad31cftjHxE7rIEvKDZCp3mENW/Tob8k47OygexQUyqa5OBaKdvvyDQCFh0tk0Cdd3tGzq7Pvu2oc4a7d4iA/iTCtqvMEIAePcczSrg8zpkDIkzg8SwEh+rfPrcurlV81gR/EiP1/DUVc/HKF8stfpmzBOHr5C4177aAxpaG00kE7l+sVH762YL35gQh/6QR2aSOk5pAtRRuoRwx133LGbV+LZYFhL9a91Qj+qlzLV13xE1DwrE34wpVmVnx9uQN60yUZIWmMGiYeFtdsPSSC+MLe5lUZ+6kwh5RRWXBgZIwceeGBBEiMOxZcfY9Fu7eFsAmBiHTcflSu+tpEVfpBCvL6jtfXzx0itaxuuDtpsM6K+NlTGEcWRvJF1bSEL+ulsoGiwEes2LO5tSoxXWNog2ojIU33NpXV5uT/yzOvcIjBS8ju3Vc/cposAIW+3TCiuXLmy0C7QeDgKItTkE892uhYjixdnURXO0Zz4cQj5EEbS8EcK5Gln7qiTZiQWRULesZjjMMdZNBgXXnihZOkSgUkRsNAZuzHOJo08T4HIEe3zIYcc0h1Rh0Z0noqbcbZOvXzHerIMEC0aP1pWGjMaw358xAgRoTBA8BxHe2dC/v24FnbY9P2RFqTAMT+54FibjOjH88yflpJGD+n2uUsaV2HhaAgPXEWQ1Fldwj+uxglS5ZmpA/MNhKN/slfr6s0VrRvFADKlndINchQxtNttmCN0dUbCEXIkpw3XdpsQ48TGog1zTymBUGprP2/hrUNOkabDDjusIIkw9St1bRwkSj/CgL/+RfxpefUdv9bxhzVCJ3+ErA1v72FEa25cObZHguHcJ5ryZM4gPwSyH97maTNhU+tkxabWySO/No57pN6JkHtmKjYNSOkgoutHiwa1VT/pH/nYaNjMaIM8OdgZ15PV13yQXvxwtL42gMi9+7PPPjuCuqvxijQr0wapn7/62jh2kfPPgiGQ5HfBoB9dwY5+2HbR/rC7tiCxO1q+fHmhGaAFiGemKBYjO2U7f8d0akoAWegsTiY8DUqEWRAIXP7S2pkj1tIRPkceeWRBimlBCDXkWFi6RGAQAsiuscTW3+mE06FB8ebCD1miiRnmLFLqQyPGjMDYRjDmouyFyIMWFumgORtWPk2do2D2nEFoEIV+/FprQXb6/mQM4ulHi2gCae/j6Lwfl62vDTizBBpEssEmuY1n445gIK2O/NuwuHd0j+zYZNOCDiN1+lGdXCPtdK/ydySPDDKrQA4RU+VFHmQrxQCtIYIb/u3VT9xutNFGrdfQewoHeGy66aaFdnJQRPWhTRbG3MWYhr3nQU5c/TsobJAf+S5PJ6X6dVCcbbfddpD3QD+KEVptY0q/D4pEYxx9yITEuyv6n2Z+UHxktO9PcUPbG9ptyhnP/Xjr8kwWyIdmGrG1blIoUfJEPrDSD0w8h9V32bJlET2vC4RAkt8FAn5UxRLYtAYWByTUIo9MEIDsndRjgw02KO0zwX3gKk2LYyraBnG8POFoktaCbaY8CAFhBAAhbfFCnmk0LJrC7MgJpv3337+wnUSKaYTaBUO8dIkABCzwjtZpmtjvO17cfPPNBc2LszlDaCdzNm02fyqA7NjguV+Mjv0l8k6LOlX9kU4EBRFp4yKoTCT0FWeT7HRHHORAuCNxcoXZCDlh3gvvO7JJGcw0HIMjZfBVz4irvo7/mc2F36ArWaceG2+8cZFnGwcZUdfWteHTuadE8M6ItrHj9SNLiGnbNvVG9GlDp5PnVHEiP5raflyksG0P8w4aZ0f9g0gXhUUb3+ayn+egZ5pM8nqLLbYYFLzOftYKGBkfzCXaDGxe2jpSlrCZRWSZS7RxnQpZiyK+dYtyJuLYqFiHkHbrVq21OPVotb8Rd7pX49V8gAUzIlphmzvjPvKwwTRe+/WFITkT9WVuYy2MdHkdLQJJfkeLd7+0eX+mQWN3x96NwGYjTeiwbaq1duW3zxZFgpuGxfEUASPSNttsU7z1vsUWWxRHwbRCEUY7x1yCloxQo3VAtgkKhNkkV67jHoQbke4fbSojXSKA+Ngs0QYivxYu43O+kPHCLNu/yRwyxzZQPXwhZvny5fNVnXnPVzuRRNepCnMixFm027iO2L04xebRfHciRC6IQ0sob9pN5dAIIq1kkPDW0ZbRktHwkQ9Iq5fwyCzmURFXPvKM58mu4pItiGobzxE0cy19SPPIHrgNn86906sgU7XWYuzQSKovnOShnuE8z9ZFXmRvPy/YOrlDTr034+U07eeYX/TjU0TQRiLwXnqjSe3HGfQsv0H+M/WL/LQNuW/zQQyZJ1gn9KPTliDyLcGUxphhH2z8ILo2WpwweTPFYJtMQ298urfZHfQCmzRTOWPKhpApBAJsDaUBRn4pgCK9st3368v8gRKIuYT6kiPWZXHTjR6BJL+jx3ykJdpl07z4JJDFZqaFE740t2y+EFtCxkIgP4tbaIgRFtplx5R2v+y0aEYsDpxdt5doLHTSpksEWgQsKBZmJwvsHWe6ULV5zubey0u0TxZix/m0S7PJb6HT2tw69re5mKouXpBCrGjezN02vvmL4OovR/LCkBpfR0AGaHHZ1JIXsLNZ7pdpU3HUUUcVBEdcRIImVZk2zfLkbNoRIJrOfj2Eh0NsbdJpiuUd/nGttRZ1trlX7/CfzhWZ9IUH+aort2LFiuJUQpuZO8iHtk8ZXt5DlvjNxjFpYIbC/lqe/bxoypWpT4WxKWZDa6z2yw8SKJ44rtNx2qMeyteX00kzWRz4WzOsBYPMEJiLIIrWDjbZlCvGmxc1++XT8Ds1cEVKo1wvTlLIMPXTV5xPzFmXjM9Q3ET86Vxt9mzinEbJzwtttOdMaLwTEHmoL3JsXNBYh7+reqqvtdPGnl+6hUEgye/C4D6yUu30vc3KPrddUNa1AoSJIyaaBtoZkzvyoKUhrBxVEwzILgFjYSCsvPAS2iO7YdqeyRaxyDev6x8CFiULI/MZmyeLV580jQoVL3ciODSFPgu42Ikv3BAKb5ybn05m+LUOYUIQLNrMGXxSiv3/oLhtOvdMDpgX2Gj7MkE4Lzc53vbJNP0rrivTKlq+iOfq+8FeJmOihcSKqw5sQ0844YTiJVx+rVNnxAz5RUpsnJARdWnjzeYeyWEaRsurnuHIROXQpsofsVN/JBn54dc6CggkjFxs/Yfdw81Jmk0LrSECNiwufySYhlN9HKnDmf9snPnIbta4cPIxSHbDgIJkOuVYK8xr2movB1o/JktH2RIbYWYLU7WJwkc87Y9+cjV+jA9jf6oyB9WHSQVtuRcd5cc5qfLZVXnaJEpHI7/TTjsVn3STZqr6SpNu9AiMDfkdfdOXdomEr521Y0mLGPskb9ayWURE2YbR1LAP82yxc1wTz9BxhCmMwPViiklMM0Or4KUOpMQVSfAJHoufXS3hRsizUyMsvPVMa0L7wzTCbp7gU0a6RIBWzeIJidAE+YyTkwZkx7h15Gx8ijMKZ8PGptPYt+FDvkZR7lyU4SjZwgsvx7RtnjSevl/MjAOpQhJoWsXRViRPWkSStk8f+OqBT2chwb4EQw6QL8yXpOPIGTb/iBIZwC8cMkA7KW/kyWZCXgirY+6I50qLTFaRQ460lUdW2Fh7B4HdKkJBHomPxPjRA3aWnpF78ga5tPF3IkXWCXOKQKa57zv1dySuPCSvDbeZ90IvDBy1R1ittSDmPk/GFMwGgUbPqRiZa+Pks2DILnmoHsi9zb++iXxcabXJY7iaD/zCMRvTDwitLzsg4PpAfH0VmwTxyVb9S/OOYPo6giN55dNOy1+8Qc4JIRyYcojfxmEzS4vvKxo2M9okHNHUl4gfuc8vnPLkxfXbtGzZsiIf6ZxpJl5OAAAQAElEQVSsaJN0+k2brEeeOZpV/e8rJDYb7GblCUNl0IoHIbdeeZnOhsgmSPpw6mceO82hpDFPIsxVXuQQvNSDXzjYWDudTpgP4U8JRFbpd/1tHKovnJYvX17U1diQr/oar219I5+8jh6BJL+jx3zeS6SlNREJArtSiwGhZ3EjPE1gQsBOXVx2R444Ced4puViM8X2yiJJyDtW8o1JCyrtBoHjRSBHkkiKcNpeR2+Ev2M2R5rSEBzMIXw6x2I47yBkAYsCAUeTyA5SVGstFnobJWPHi5PIDu0KO2Caw1E0CtFxumEz5wd5ELR2wZunOsxJtogDTRn71lprMdcswG3mjtFpJeFMo22+wtsmg+ZS2xEFaXwKikywEbBxpYG0uCNXy1YRGFiZ8zvvvHNxskTWmPPScmQM0kRm0OSTNUiJ780iUF7SQjrE5WjT1IPsQlzJJeHqSKMsDGlCsvWJvNnz+pEC6bVDf5FHe+yxR7EpR6j5qzOzCHjYmIvPsf91BO1UCgFGoBFUYeSfkzOEyRg0TvlzCCiiKwwZg7Vw8lC5tNfscB2DO+7WFnWGL9IkDw7J99WMWmtxtA8f+QnjpFV3pEw8tsb6ABGHN0zNE3E52CB3PvmGyEsjD+YLNhz8mZKIy+kHGmO/rob8WjPgLCwc8wr96qtBxoPNE8JnjtpQaVOMGWmYLomDxBpj0vMPBwdtQirF01f6yIZL/2gPG+WIrw9p8ylQKF28aCYP/Wlds4nTTtpdY4Im1oYr0ruSI7BD7G0mjBv+HHLPlt0YNb6YWfDnjEmab8TWWoiw8+dsSNi8G+fuaaiFG1/WT9pnmFs/1dd7NDabxoa5JY90C4NAkt+FwX1eSyVkCQSTnEAnTCyGnjkCh8bWPUcjQ9Plnus/m9QWFwuY/By/IdEWJYuXoyTpLQZ2+IQWIaaRNBGONwlVWhfx+adLBCBgEbaYGXfhjFdhnA1T+NNk8ZtvZ+E2jpU77NNO812HmeZvkUWS1D0ckjsoP+TDizcRj8YMoe3HReZo5xDCiEsbiiQgauKTERHmE1D8OMf2iEmEuepvGlb3PrvWaorJGf7hnAREOG20dwpoSckam2+bE5o3xEJ54ZAtGtC2zjRutPg2B218372lwYwyXaNdiDDZxg8hR/aiDORTXYRxiFMbDkunZLSIwp2QIYk02ZGHqx/lEB4OtoNscpE584EcFZd2U/lMhLzQJa/WKR/RjbbBjJbcaVxbB5shfSLPcMhhm5d7+PsSCzKI1CrfWsKMBrkWJ5xNUeQV1whrr/qWZtRGUzwYW2cQSiS+jYscU8rYZMX8tK5Yr5BScZl8yIdrNeLCKGz4h2vDbRbCP67ScBRFsZ7a6CDc/Dlf/oi6S9eGq6/NvA1RjAHKIn3iaynSp2sRGO19kt/R4p2lJQKJQCKQCCQCiUAikAgsIAJJfhcQ/Cw6EUgE1kYgfRKBRCARSAQSgflEIMnvfKKbeScCiUAikAgkAolAIjB9BDLmCBBI8jsCkLOIRCARSAQSgUQgEUgEEoHxQCDJ73j0Q9YiEVgbgfRJBBKBRCARSAQSgTlHIMnvnEOaGSYCiUAikAgkAonAbBHI9InAfCGQ5He+kM18E4FEIBFIBBKBRCARSATGDoEkv2PXJVmhtRFIn0QgEUgEEoFEIBFIBOYGgSS/c4Nj5pIIJAKJQCKQCMwPAplrIpAIzCkCSX7nFM7MLBFIBBKBRCARSAQSgURgnBFI8jvOvbN23dInEUgEEoFEIBFIBBKBRGAWCCT5nQV4mTQRSAQSgURglAhkWYlAIpAIzB6BJL+zxzBzSAQSgUQgEUgEEoFEIBFYJAgsWvK7SPDNaiYCiUAikAgkAolAIpAIjBECSX7HqDNGVZW//e1v5aijjipbb711+da3vjWqYrtylPvgBz+4fOUrX+me5/vPBRdc0LXxUY96VHnZy14238WVH//4x+VhD3tY+fSnPz3vZWUB6zUC2fhEIBFIBBKBGSKQ5HeGwC3mZBdddFH59a9/Xc4444ziflRt+fvf/17++9//lm9+85sjK1f7/vnPf5Yf/OAHBRGeTlvV8Q9/+ENX1+nEb+N88pOfLN/4xjfKeeed13rnfSIwEgRsbI877rjy1Kc+tSxbtqzc7W53KzZ+r33ta8vRRx9dTjnllK4e97nPfcrtb3/7LnybbbYpl73sZYur+Py32267Ll7/z8qVK8srXvGK8q9//asf1Pnf4ha36PKUT621XOc61+me73SnO5XNN9+8fO1rX+vSmYvmyXOf+9zygAc8oNzlLncpu+yyS3nNa15T1PW73/1uF8+fP/3pT+V973tfefKTn9y16d73vnfZa6+9uva8+93vLmeeeaZonUz54Q9/WF71qleVnXbaqSv3/ve/f3n5y19eYPLKV76yi+fPueeeW77+9a+Xpz/96eW+971vV/6OO+7YxX39619ffvSjH4m2liM3DzrooE52rhV4sQdsvvjFL3b1lae2qQ/cDjvssPLlL3/54phrXsgpeHznO99ZM2DI05FHHlkOPvjgoXKK7PvpT3/a4fnIRz6y3PGOdyz3ute9yooVKzo84dJm/de//rVTijzxiU8sd73rXcv2229f9tlnn/L+97+/PPvZz26jTtz/8pe/LC984QvLz372swm//s0555xTPvjBDxb5GnfGxiMe8Yjylre8pevDb3/72xNJjAt5weoxj3lMucMd7lD09zOf+cwCu/e+971DZesRRxxR3vjGNw7Ew5jmlE3pc5nLXKbc6la36sbI7W53u64c7b/HPe7R3Yt385vfvFzhClcot7nNbbp4rubSJz7xiSKNONy1rnWtcsUrXrGLo6+33HLL8s53vnOiTXHzl7/8pTz/+c8vp59+enitdRXnmGOO6cZ3zN/HP/7xRT/zH7Wyaq0KjpXH7CqT5Hd2+C3K1CbqjW50o3KJS1xipPXfaKONOm3zKAu91KUuVa5//esXZU+3XAL92GOPnTZZbvN9+MMfXrbaaqvWK+8TgZEggOwgjhbKO9/5zuUDH/hAOemkk8rb3/72gpQKO+uss8r5559fkLJPfepTXfib3vSmco1rXKO8+c1v7p6dWgwivzaFx60i1kjGIHJ45StfuXz4wx/u8lBurbUgMO4/97nPlUMPPbRE+eLttIqgIheve93ryuc///ly+OGHdwQNOTz11FM7zP74xz92JzaIHpL8kY98pHzmM58pL3jBCzqSY57aqIqMND7jGc8oZ599dheuXOEPfOADy2c/+9kJ0qn9b33rW7tNAeLytre9rSv/Yx/7WEf6vvCFL5SvfvWrslzDwRf5gJP81gi8+AGpRtCe8pSndHJAm7VNXyB+H//4xyc2ABcnmbjYDCCE//d//zeU4EVkioQXvehFBR7kVfi315/85CfdJggu++67bznxxBOLPnePFB9yyCET0W0wEDP1hRcs9BnSi5TbfExEvvgGHjYQ6jwMjz//+c9F/xp797vf/Qps9IsxeeMb37gj1mev6i9Zyg8R3mOPPboTtOc85zkFFvrbZu63v/1t0Z/GhPits+mDx0c/+tHyq1/9qg3qxvtDH/rQ7jRO2epjrBoDnmFiflzykpcsj33sY7vxxR8Bt3Ygsp5hYgNhc6M9/DjjWHr3K1dtDm0WYFZ6/4xp+MLVGOwFF2PHhgQ2O+ywQ0F25YnQ3/CGN+yI/bC+7ueVz1MjsMHUUTJGIrD+IPCf//yn00ZYMNafVi/ulmbtVyNw/PHHd+TCAr7zzjuXzTbbrAtAMGnPkNxNNtmkbLDBBh0pvdKVrtSF9/+Ig/T0/Wk9ETT+CINr6x70oAcVhKb1i3uaNuSVNuv73/9+p2HddtttOy2kNIjH5S53uU7r9uIXv7jbsCJDtGw01jS0tLhRZ22jPUTYLn/5yxdkY/fddy+I8H777Vdufetbd0Ur92Y3u1lBjO55z3t2fggK4kejtvfee3faaeXXWjvyjVjTWHeRmz+0kjTn17zmNTtNXBPU3aovgkPz/LjHPa7sueeenbZb3htuuGHZZpWGXX2jbl2i5g+zLBpFBPn3v/99E7L2rc0DLeNpp53WbTb6MeDxtKc9rSCKtMm0lrT7FB408AgwvCLdO97xjkKzStOrnzbZZJNSay3aajwhohE3rk63nOJd/epX77TL4R9XmyX9Z7MEU+ZuV73qVbvgjTfeuNMsy1cZPP/97393mv9LX/rSnYZUvxm7wq573et2Wmh95rnvaJZt+IxPm6c23Hinddam1j/uEeEDDzywO/0wb4YpStTTZo5GnEY60rdX+NIoM31r/d07URBmA0nDy691iD2CrV9sFK52tat1wbDSJ8aV+84z/8wagSS/s4Zw/DKwi6fNcLxD+DmqIXQcHfVr+49//KMccMAB3bENgdiG0xTc8pa3LBYXu1umAxFOA2IiExSOk+z+CWxClNC1Y7fbFi5/R0qR1pXmwiJlwbOwteEEmMWaECRkTjjhhE4TQotgN04A0RJZoLTR4hKO5kD+BC2/2972th7XcjQm1772tYvFkdBkg2zxUv/999+/O5JTNxoniWnD7O5h8ehHP7rTLvHnaD8cJ6qvI9S+5kGcdInAfCNA63mVq1ylIInGYr88xMq8skAPIwKRBrmM+7iae0jx3e9+906LG/5xRYLiftC11los6Eg6Aqy+CFkbt9ZakB7HycimeW7ecW0897XWznRDGHljHmrfFltsIXgNh+DQHtKuIY5kCbk0qHxyDcFZI4NVDzSpNgCInKP8D33oQ6t8//f/wgsvLGQVwkZeOnX6X2jpyKS8mRSU3j/1d1z+rGc9qyC0SGUvysQj+U4rSItIEyltn0whgLSxD3nIQ7oNSa11Ir0bsu2ggw5y25nAkXPGBdKGLHYBF/8hI5mcXPw4cfF+g80GQq9OyNtE4KobZJYW1PrArfJa479y+IeMZrJCLjuVuN71rrdGXA9kL600Euo5nLJp1m3ujHGa4nY90a7YNEWa/lWe6uNUtB8Wz7XWYt5suummxdoS/oOu8mv9mcNZq1asWFFskPRxG+4ejsau+TUof2vhoLFTSpE83ToikOR3HQFbLNEJZza9hI8JQytDWFq8COlohwXDQnPwwQcXx0COjYRbdCwOdtQ///nPi8WQ3R5iStDKx/ER4ceeDHGkCSBUHdUQzoTYrrvuWt71rnd1R68WM+VagBwtqpNylBnHjOpjF46oE560Duz7LJie1U0d5GHxtMg4SlM/5WyzzTaKKLQd7LdoeTqP5g+7QwuYPH/zm98UgpbwdxS1fPnyQhtEc+N4C9GmiWHD5/iSRphgZeemvZ7hRCOhftoL96a4vE0E5h0BRAQ5s9k0F9oCHSsbv9yXvvSlzl7VPGrjTHX/ve99r3tJddmyZZ3dqLls/k+Vrh/ueJpNp/Q2nf3weK61dmYNNK20flNpvMxp5CXmf+TTXmutnWbYkT+CXeuahJDMhBEHJwQy+KpIeAAAEABJREFU0qsvskjWISE3uMENChloEx9xYCodUwpELfxdKQbky4nTkh9yhrKCdpIWnJwlj5UpbevIWfLIsThSigzamNtMtPEQX3g4Lm/9yU0yK+px8sknd2RMO8jLWtfEJNLWuqa/ujmiZ6etH8lQMpcyJdKQj+Qvk5oNV2m+w9/VWFUHDh76RJ2R1P74JVetCxHXlWZbPvBAmK0l1h5rjg3IILMc8RfC6V+mMtYIdsROPGhxYRj1oVgyL7QfuQ5/1z5W4tGqC0s3cwSS/M4cu7FNSfuz5ZZbdja9hIHjN8LUjpLWgzY4Kk9zwBbNjp8WxC6adsOLJHb7hLH8EDzClq2TXakJagdLqCOo8rDbtbu1CBDkCKQjTJoox6RMCpQrPeFNaDq6s5AQysIQWMIPYSe8HW1abNic0d4owwKPsNtFO86Uv/xoYghx+Vi4aJZpYTy3Tv4I9RZbbFEsGoQTAdvGiXtCNoQp7bZ6iEvLQbjTODhGdFymvq7DjjUjz3m7ZsbrLQIxt8w9m7gWCC+E0YqZU+Y04tUuvG3cQffyMy+9MIbUIcDs2pkOmP+D0gzzs2jLb1h43z/kQt+//2wO9/0GPQc+5nA/nDb4Pe95T3FqhKg4+Yo4TroQOzILmXOaRingmDriIGLqC1vtDH9XMpXsdZztWNtGmj+H/METMZK3UySylb/w1pHPNtfbb799Z75CDtocqHcbL/DoY61+Trl22223Yj0g59mR9uvb5jXongJCGdYPMtlp2FlnnVUoFCJ+4AFz9+Hv6qUvY8qYtIkgh8laceEnTjjlILxPetKTupcjKWn0hXDKF+QQcafhNTZtPihVhI+D09fWBH1srWB+ow9C4aOO+sVV2zn34WBl/dY2p6SIMpwiPK8zQyDJ78xwWzSpEDyVRQJNHpMmBAf/OB4VXuvq3T0BS7vJVEIcjomBIzsaIOSTwKIBpulkoE8zg/yJ6+jIDrbW2r1oRstiYQkB6ziQoBLX1Y7dPUcIWmBDU+CYSbl2uyEUkHB5iM+pjyNNiwUC/Itf/KL87ne/697IFd532uKoVBpEQN37ceKZULJwMf8g6Gll2KnRNtgsEGDaqs7SqFebHxs6wi4cIiJeukRgLhFgTiA/mz8vBrkPh+Qwaaq1FuMQuTOHInyqq8XWUfJLXvKSLj1zJvMUaXKyNFX6Nty8jvnh1KUN69/XWgvTJKSoT+L6cc1pfmSS6zBnzpJPCCQtdBvPy002zDb8ZKXTIeGIMntNONhowxBpJI98ujGUCeQf4oUEtjJWHjTSFAHkMQKNfPLXX06+EGP5UiIghTbWFA6In3gcAkkDKq7Nu/iINNnILlr9xOPgQdYjTp7DUSAgkUinMWAzRKZZB6bqj8gDHpQgvsgReCCbNmCUAdYPccl2xJg8btshjFLGeHLv9E4+SDztu/jqLoxTZ6SP3av8yG0mNvCg8WdD6yU5eFD0OOnQL/22y2vUzjqqvyhL1E//OqnUv044rS/qpF2uxm+MJ88crLSf0ghOTOusM8LSzRyBJL8zx27RpbTwENCuk1UeSSVY+kdHSGYsXMjjS1/60u4NcUeTFo2YyG3eyjNRkcNaV5PrNrx/T+iZ/ISGMOkJaQS51uHpkVNE3HESLchNb3rT7jM18ug7O3F2vY4xLSLa0o8Tz8q3MBC2tC00DBw/GBH42k2YRZr2asdP6IWbCvs2bd4nAtNFwPhEACz8tGT9dEgfP+PPmHY/HYd02lAiHDShTnA4pIMsQHbWRfurfHNBfacizhZ7GkVl2cxOVl/H72QH20rzcVhcpz5OkhBXBLYfD2Ezt8mrCCPfxGdaou2cOjG78o6BDbC46ssMQbxBfUAOypuLPkDQaq3dlwjky7H31R5X5mHy5vQF2YVEu4rr6kU1/eszceJxzDPUWZ/ZvPMLp43ie9YfZCetMzKrPvwnc7SW1gjrgzpwNNVO4miE3Usvb1+/UQebDX6tC4zJd3ggwU4ZmUpQlrRx3VPQwBh+npFwJnbML3xBQT2ktXkQh7ZUvIV0lCvqDQP145wiMNOwkYGl+tH4Oym1gdHv/Fqnv2qtBaawasPyfmYIJPmdGW6LJhVtqEXBjtoRnYWHdoIgRd4IEI1B4jzTRNzkJjfpvoHIps/RHiFKK0A4eiGDMPUGr0VPXJoE6cSTl527RVh+Fi02Zo77CDkkUTxXcQlRZVuwxEdgCQcLijBlONZie2viq7f4UW95hFM32pzjjz++0LSEkJAGBjRI4qqP9AQpIWu3TTPhCE0ai5R8hKmXvBz5ElzwoPVio6Z+jh0JK0Jf/cWXN+0H7Q8bYlrycLBXh3SrEYA77IzTQc5Y0N+rY4/+rxdoaPYt7LEhG30tpleiF9IcrVr0jVXjWkpzzfx333fGrLlrjhj//XAnPeYNjZX5Eo5m0eYR0XMKoow2bZQHM/O9DaOJps1CWpgUmSvmkjjqjDwhBuaieW/TrU1OTYwX8ZRHJiE75iINMdMOL03RQpI7yo32mbO0o9IyyaJFpFm1USZjonzz3vwVjyMDvL9ALkXb42rDL65w2nb11S75k0U0uvJWD07b1Fu+HPmArBtj6h/5urKfFZctsHZKDxeysN8XZDryaiOCdIvreN0zORz9pK7wUG7gqB611kKRQTbRRiLAEW5ceIHXVxvUQ1/B0UZLPVtnM2CswEO7EVBY+GyezROialzA2pyWl/LD2Uwh8GS/sUyjLZ5wddY37jlttAGx1mh/Ww/4UH7YXDF/I/ul4bRfPtJTsvAb5tRPH2iTdIPiqR+ZL0/3bRxtdWLCtM9mr60jcz1t8oK1sSp/mxrab/PCpkd6+ck37j2nmxsEkvzODY5jmwvhajKZbI5WHOXZhTsqMrHZT7HTYr9nkrNpRUIck3oT15cbpH31q19dHNV5m1pjLVoWD0dXXsCgTUFuhRGyiLZP9xBmjhlpIgjEN7zhDYVQYSpBkDs+Q0YtWISjHbEjR/a18vaNQ0e6tAJIkiMk9ZdOOcoLR6vDzMBnfRxh8tcmixBBqp2+2ehYk7+jJPUkuCyatDgINjMLi76yCX51p/VFgAhzdbfYIMlPeMITCoFl0YARwc3fYpsCSw9M7iwA3la3SbBIeHknnJcQLaiIxeS5zF+oupkL6uIlTGN4/kqbXc6bbLJJ92KpI23z2obLHDJ/HbU6LkW0ohQLrjlhnGsXwkFrZj6IgziRF15EMsf5hTM/kDHkkOxA0iIMIdVv8keazPmWWCM5ynJkbz6SIwiaOU0uIVnmqPyQGMSXrPI2fLSJ1haxMNcQPXEdeStX+UyTvJTqJAgJtHEnV8SjXUSUzWda7SgfVsgIOUL2IU3qbjw6tmbmIX04MqrWWsgQ5QlHcmCG7HnJ6XnPe14hG7SPMsGmARlEDBFccs8Y0+bIl9yweYGftiDZZI76kmOIETwiPnmNyJGHxirMzzvvvMI8BXZIt/LVxVhAdPWb9zy8uyEf72qQ/TSM4qibePpH+60dcJOf8QA3slHacOpYa+2+xQsP4ZQj1gBz2zoEG/nrS2PGuxpkqTxopJnkKMPYU1/10C/qYpOD2KozzLXVhkj7kXR5cNYRbSbjxbEGCkd4bdS8+G1TZgza/JBB0oWT1vjQ95QdTF70UYTHVf7qaMNhfGkbsh3h6s78wlqpf8Jf/vrIJkC+NkrCbYDMW+uPsQJDbYeXMWz+UsREPnmdHQJJfmeH39inRirYdPnkjcWFEKPJsvAQCgitF84smBY4hJVdGmFIABAeFg0vgBDoyCEhaGHhxw6W5pcj0AGCDBJqCLEXwJgY8HP842hHOUijxZowQywIAAsDwWYHTHCot2NEX1+wIDkashD4AgWNkJ208lqnDtIwk+BvcaSdJeQs0kwcPBP0TB68XKJsgpgtXmhvLEow8xkeRJrwpe0mkBBsR4W11u57pIQjIWjBVGfC20fuaVLUId1wBGxskEqCvdZa9H04C4BxOjz1/IfYuBn35gFiqF8RkPkveWYleNEV2XJS4aUsRIuZjvmJnLAZjJxrrcXXABBGi7GF1vwwh8RBUm1+9YO5zC+ceWr+W7yRBOVEGDKqbGHIk7CYjxEnZAgCYi4jR+YaWWODrWxxESLzkuwy981t+SHxZIz5Zo6La+7S0CK85IsNqzkpb+kQU/E48gRO5q404mgj+aS95n6ttTixQkgc39NASxtOvZASG3LmGRGu/eQEkkX+ahOZw5TBRgMphTEtMbINxzABkDclgnrDj6yThy8EII/a1v+BIrLVxttmgPw2BuDGUSboH+M38DIGyDOktH2vg4lW4KxugbN78k5+NOZkJeJMnqtvOGsIrTBiTGEgXBqEzdiCtXRw08cw037YRx7GCZxsHKLO6hF1ti4giU44jB2YWL/0feRh/dKHSLtybaSEc8i4DRRCag2TT4yfSG+NQ0DNISSbXbSvWUR4XKWzftkE6Sv3rcy39plz2mlTEenkb92UBl5eGNdu9YOzOmuXMaftNNs2djAxlyOfvM4OgSS/s8Nv3FMXgtIiYJGw4KiwhYugMqkIZtqVmGT8CAjxkE2CyjGlSWnS8iecCQ0C3WSkaUUShHEEu3KlM8kRHBNbHPmHU4+4d0Uya63FCw4WN3kT+p7la1ERL1yf/NpRs4ETLj6nLurOj7OoEDTqLH95ElgWPvWTBilXb4tBlKEN4sCSsA1/7YITAQ4PCxH8XGsdbqOsnHSrEYBlEIfVPqv/Wjx97cOmaLXPwvw1LpAZGygLu80cTenC1GbqUuFmDBqPxrwxi1ga5+Zum4O5L0448y7muU1y+JuHbTrkMcJc23Dzg184c6kld5GPuYmYmDviqi+SEeVHPFcyACERjzM3WzIrDmcsmXvknXjkGzJozgtvnfLJnIhLHpAP6oCsmNvwkA9nHLTp+bWuDYezjYW2RxwYKU/e8gl/1yD7/IWTNfzDSRv3+hapFJcjayPMtQ0XT1/BSxhHpnsJOdYDeXC11sJPXLIbHuQkggwLecFHHlzbXun5ta4NN7+Ntehr8ZBiRBaJlD6ccvSteojHRZ2NSXXRp/w5Y0iaSK/9/MNFuHTaHf6u2qevIq0r/GNMiMPBWFjrjB9h4WCGeEec8Hdtx/+g/NtwYwFWxq605q96UlwpM/LP6+wQSPI7O/zGMrUjNC8AOBqzS/c8iooqx06Z0b5dNzI63+U6vqIdJBhocWkGak3SOd+4z3f+bA5p1Gqt3Y8DjKI8L584thzkmLNY2GjhaOOcFrRH+fNdv8x/pghkukQgEUgE1kYgye/amCx6H3ZKdtoWaMb07KdG0Sif+GH8z5xAuaMgBzQ6jsnY7zpiopkYRVuzjPlBwHEsR9PKJnx+Slk7V/aEtFI0TJM5b9ZLzQSG3aL7dIlAIpAIJAKLC4H1hvwurm6ZXW0RQHZDHCN8Jg2zy3F6qR3VKTOc46bppZx5LEdmjobYAzomm3lOmXIcEHDMzDkudvQ5qjrR6v5PF5YAABAASURBVDqtYDc5mWOfqk7s+OLec7pEIBFIBBKBxYNAkt/F01dZ00RgySPAdIXzsoeXj0bVYHayNlHKHuaY8Xh724ssXjCiIR5V/eagnMwiEUgEEoFE4GIEkvxeDEReEoFEYLwQ8ILKONTId3B9tcDLd958R3y95DgOdcs6JAKJQCKQCEwHgTXjJPldE4/14slHs32H1mdmRv0NVeV6kck3MecCbL8y5HNDvls5KD/fmvSdX9dB4X0/3y9Vv4jv25k+zeaFJ99l7Mefy2c20uyWZ2qjzd7ah9+9dOhboXNZt/nMC8ZsxJXBbtx1XJxv3vpklS88MHXw2StfMxiX+mU9EoFEIBFIBNYdgUVBfn3w2zf0BjnfyPPGv6Z76akfx9FphCMxEe47esiCD1/7JZjwj6tvAfqINa2Pj0yHv49w+5yW8har0y7f9bSg+7W2UbUDlt6k933NmRK8fl29FOWD9344ox+GUPk8le85egmwH95/Vj8fLPdBeL8cJNxGwYf3fVjeR+f5zZczTn2LeKbfkfWhdMQdWRu2GYj2+N6mb5naGPCbrzZNla9+8Q1ahB2+voHqG8tTpZtN+Lqk9XUHml5mGL6X6rNR65J+IeLaQPhGN5lFU+1HHHyE31cqBtVH//tW6aCw1s/4hIFvuMrb93H1m7yFtXHj3g8IsKWe6tNw5FCbtx+T8R1ym2RjJPJrr2Q3jbz6t/79e3PZN1KNd/X23VY/7uHrNMPkEBlJbvTz6j9rl7ytC/L25Rn19sML/bg2z74v64Vg37X2DWG4+zqOH9npx/esjeRPbMb5hRPmW73KHeZ8wzlko/llbPiWr+8eqwNZ7EeGrLF+1MN3Z4flxV/7/Kpf1MHmUP18q1i47ysbE5QIfiku4rmqh2/9mkvi6mN4nHbaaQXevk7kO8bChjnvdshHfhxTJDKe/b00NqjWc/0amOofYcMc+aMf5dc6dfINevVq/fv32uo9F/n7HrGXdSkhfD2mHzee/VATpU08T3WFm/VpqngZPn0EFgX59UFsBMck9TKK71Uisr5J6McKfLPWz2GaOAaJOJzJ5cP0vjdISBDa/Dk/SuAj2b4L6WsBBIJfEBPm268Gs2/v+WqCj9x7scoLOAas71FOH+Lxi+mbmV5Gm2pSz3XNYellvNlMYkKj3XwQ4ASob2r26+vbvBYl/dYPG/Ssfl62autn7PhcnB9iaL8lOSj9bP1oFP3KnPEcedFs+8WieJ7s6puZ8qBJHRaPQDZX/Dyp71sa29PNf1ies/E318xtizsSo342tLPJcy7TIgOImR8n8I3Rucx7PvIyNyzCxg0ySV7ZUJBniIGTl365xgP5CPt+WDyfcsopxfdTxUFA/FKWH1Tw89NeThyUr7Tmq6/OIDieBzlE0ZhXhg052e5LGuaC79uefPLJg5IV6cht8QZFQPZsthESRMxnEOVtMwAnMkPbB6VF6Gx6ENNB4fykZR/u582tJ/JGJq0hfgyirbfNgR/+gAXbcv3iV+N8Q9f640RBnn2H+NsQy7MfhkzXWot+UDZ7dP2BgHomt6SJzTR82akHAdaHCLivq/gFTXjJ0w8qCFNHpFAb5WdjbfMXmFAeGVN+UMJVHHUxFnyb1i/BKZ/TD37JTf3gD19YUTaok6+tKNt6bF2WF+KpbxFuzzYK4iC28rSpYX8PRwRWHFjZMPmudZSPxBqzwo0XhJ0Jk2cbbZ9VHKQs8Ot62hbtVWbrbD58/QV2vjqEZMNN3n6oh7952KaJe/XQfpvD8Bt29St25rJfaRwWZx39M/oqBBYF+WX7h3yuqm/3HwnxsWcD04RFig3UWmtp45lIng1AH6m2YzQRVqxYUUwipEmG8hOv1uqxeJbWA82igeqXjkxG5aqPsMXsal3d1lG3odaZl0tI0Wrq86i3vtBXta6db62168ta1w4rQ/7VunZc+RsTQ5LMmXetq8evNslUey1gbXv5D3O11im/iesHBCzUFmCLJaF96qmnDsty3v1rXd1m8yrcKLCebsPIBfUapzoNq7tFGlFAdikCkFUbeF9BscFAsvpjyc++IkXMZGzw+hsn8SkFbEiQIMoE+Rk37J6RCs5zv17SyhtZYWLVD/eM3CgXCaPI8MMH8PZDFYgwImTMits6G3dzg0x3etGvt7g2VLSsiBiCp97ytuEi082DQf1q3sXaoAwYya91yLQ5hJzbOETevjfuVBGB1H5pEBy/DMh0CoGxIbep1i59hdhor7itk5720gbCr0aqVxuu7vFDFNrF1VoLeeXeF0xofo0BJ1cUBUiq+loPlW+zj7Qh67XW4hf+9IX+lEettZOh7r0UimT6gQnrKA2ucYMAUlCJQ9FA86kMc0cbbAIonpSB5PvRIT/0YLNuQ2IDri7aA1M/ZiIv6Wv9X3v8QIh1WHv0O00+hQX8KKSk8cNJxqM+kF7f+ZU7GzTh/MhXZXk2hvEB19L8M74QY2PIKannJri7xTmMIRjrX5speWof7bNNl/Z3kZs/yCzOgrz70ZwmaK1b2njf7Fdnc9ppxVqR0mNGCIyU/Nr1GZQcgUVL694ibGe8ri0wQSONQT5ooAknBEx8mi7x2Fby8xkw4cOc4xv1JAwI/mHxxs3fjhzWFhz32kB4nnjiiWtV1aQ2iS0G/XAaAseKNCAmqWP1yICtsIWQkCMkaD8IZ4uZn22kbbAjFy5/Wr1I66pc5clb/AjnTyujPsaGMJpY5dkpW8SNF8eWFhWaKP3k+Eu+NjU2LOpt4XScp8+FqY88OekJFunY2fJTZ/FaR9jYzSMA8hafFoZWQZ1siGgkYBXp4GCx8fOYcCf8Y3FWFxojGgrp5E27o920Y7QtnpVL+xXtJYCVDyeaWnOHo1HRx1H2VFefo7PQiHfuuecW2nG/HuQ53eJGAJGwSNI4IRdtaxAGJIGmsfWncTM2mYchqsZWG27umd9kqzErnzacDN5tt92KE4fW3z1tmjHtlMFcRxj5hzOeEV7lI0DhH9doCy1e+MVVPdXFcbv5iKREWFzhYY6rH7IZ/nHdZ599CqISz3GVH7LENIjsojyJMFfzjWlUrbUoo38iYH4h9E4oxUekySHyC7nn1zrfl7bGtH7uyQTabfXXPrKCfzgEblB+Ee6KYCL75CzZw+TDOigsHMJIYys/2tl+eMRzlRfya03RJmY1g/pev2iXdZ0shgnyZrzIJxxCbXzACjHdeuutI2jg1a892tQZizZ7yLA69SMzg6BRlqf4/XLb+DYw/fFBvjtZRvCtFThDm8a8QJoRdXGQ3jZcu2Aqj9ZfH9B6Gx/Isj61prRx2ntabCRZfGZ8cG/D837mCIyU/JpkSJljDza1Bi0CQ/Ahahb2dWmKxV98g9vPEdrxeu47k9ngNIjZOrF9M3j7A75Nh7jYvSMGBGEbNu73BA6yRHNyzDHHFDtHu3NmICZeW3/EzKJEw0AwhhYQmfVMeBA4FkACBQFG7kxsGBKECCjC6oiMdoOQI2zlDWvCDYm12CmbULcYmtSEuqNAi6Qw/hZpx2sEFgFjoXOvHupDa0FTYnFlT+hIWr2kZ49mN05YEcqEmDKEWfjZ4/mBAgtrrbUYG9quLgSWeK2Tn/y1DwGQl/RIrSNxGhACTJu1yxiGNaJNkFt8991330IT4B6xpf1QF/PBuEdA9BnsLaqetVcb2/bWWgvNkXJpQYTROFhclNvWe6p7dprG9w477FBoTaaKn+HjjQDTDHOe5o32blBtmTsJjzDj2ebJBsg8M+ZsBMnkiGMThhSTgdKHf3tFsGjgWj/3NsfIFK2x8U4jxz+c+UA+IRDCw7+9ktG0qa0fuUMjS77RJpI75nkbxzwle2j0/FRsGxb3yLOfMo5nVzLFPNRehINcIDOQFuGczbXNMOLZTy+cM7fJK7KS/JIvMi2s78xja1jfXzrlksPmu6P0yYhSP3080xwzRbCWUfqEf3tlBtc+T3Vvk25N9ZO+w+L6KWHyzmbfT/X2NwmRjpwz9uJ5qitZT+FBdlv3B8VHtpU/KKz1G3TPLILNvP63KTEGrEttXOuctcEY6BPciGfs9cc1ra/6qzfeM5ltsPXS/JQHjb0+Qn7N9SgjrzNHYIOZJ133lMhuCAuTwcByjGLHyYSBgJhOrsiFwe1FNAIGubPLrLVOmpz217E5kuLYxBGLnVU/EaGNoBP8iJvB2o8zzs8WOIuOdhJ2tKgWxlprQQwRtqi/hcdRKdKlnYz3TX4vezgupBFyXGrhQnKRNUTNsY3+s/g49rHg2r2b1IStIx9HjezE9BENaRA0i57+l7e0FqGY0EH85Es4yFMfbbzxxoUGiMCzU1ee8URw0hxFe9Rvk002KV5+dKRIu4/oCjdGjBnPFnJXBMDC4uUdQka81lmU1E+d+SuPECesjV3jw6IGN3EsjBYGwsp4c+xKA0GLq/0WL87YUz84y9diaZEO4mHR107j3FV/Oj6mobYJcZQnbzZvNiaEqnym4/QJu1AviiARtEvTSZdxxhcB8gxRQqSMmenU1NgnO51qGdcUEpQTyEqkR97IZXMg/OJqg6uscMZihJEhlBPmARJiLtCotnkrHzlCoszFSOtqQxz5uiLn/DmbanKL5owcYC9sgyosHFlGQ0shIv/wd6WtlWc4cgahFkbLK76Ns409ZYkNNoItnIOHuUwmeQ5n/lvHIl9X7TbfEZn+5tpcFyccuUIREfmRn2QkEoUAO5miRY7w6V6ZJlgLhpG06ebTxkMIa61l0Lho42mj8gfFY4cebXdlE92mHXZvbOmvWmvpn3BQ7sgrHHMM6/iwvAb5O7WzjhlfZC9nDW3jWpNqrcU60PobdzTBUb6rDaU4xoDxbzNH3jN/sfGkQW/XZHE5c89aqi7GmrXCxgxhFp5udgiMlPy2VQ1hh8wgawaoziYMCbdwNJAtuZEHosKPsEEIDKZaJye+0iHZBDESh9RYLAw8Ya3zwgCtgoUEGUMYTbg2zjjf11onbD9p9SwAtDMWIhg7ro/609jABYauFiSY0pzAqtbVecnHxoUGkzA2cR052SjIH5mFV621KIt21rO+tTiZxF4QUC6BxRkDhAOhrFxhtJEInoVy7733LjQW/Ae5WutEOyOcRsDCKN9aayE0kO8IF4aMIuMWBO08//zzu5d5Ik57rbW2jxPlEebarQ2IdAhBZh/hJ6F20o7ws4tHMmCijbRViDd8aq1FnDLJP5hYmPWVaOJbnI3R1uxC2GSOrZz+oI2Hjw0DTbkFJeada7sQT5Zfhi08AsaFWlhgY555nsw5QUAQyV79TSaSvzbK5IS0ZIJ57MjVc+tsiMU1htw7ARGuDsx1ED+nKjTHxrl6UVyYd+Jx5inZ2vrxd5JCu4pkOt1gwsQfSTDHlSlf2jfzz3hmmiUOZ11x1T6bdffhkEp1Jf9tJJlmkYPWE5sBV0fqiApZZnMIn0gPD3MPARQ3/GFlM29TKT/mCupKTogDW9dwyLDNMvn6iAqSAAAQAElEQVRgk0/u2gALNzfhor3KRpbUl8kX8i3OdF3UF5bTTTNVPHJNHH3qOszBRPnGWD+OTQjNP1lKOaBt/TiDnuUHf+OM3GrjMPlBXikyaLqPO+64Yr1q40x2D3Ondk4e4U6pYH0iC+UVaY0vfW9TpB7hr5/MK/NBHZ0QUi4JNw5pjLUXiaUwscEyh/S9OK2zlqiDDYQrzK2T4pPVbdy8X3cEFoz8DqoqgYvsUO2Hs+smAAbFn6kfu0fHvSYmIjIoH5pfRvomN00eYU74DIq7WPy0tdbVLxBMVmft1G7apDaexTEWWRpPmwL9BStHjNK08d0TVLSYrrWuSSSF952FxkJncsuXZqcfZ7JnxF79h8VRFxpXApIAYtNHOFpMh6VZF3+bMuPVNdJZnC10sCOUadnZGtLY2mTZ4EXcya7yhbFrxCMQkWFjOfymcyWAvSyDhMOao1nysko45hTTySvjLDwCNlU2drSM09kIIQ3mmXQha5E9G0OLayzGZMZWW21VvJVP29VvqbGN6BmDEaZ8x8TSRt5OdnzphX+YOCEHtM7G/6C8ad7kiXwgye4RBoTDWI285aMs84qWUTwnXttss02xuZWGX+uivkgOIiNMm21IrUPyRlzNiy222KI4BheHs7FHVhHkQXmb4+ornnnP/MnmeBC5408mqX/IICSJaRSFg3pwFDaIHazISPWYrnNcrr3qSjM5MN06erL1JYucAE6W1OYBDuSsMdDGrbUWfWv8iNeGTXYPK32LzMu3HxcBJWv1EXz74cOeEUv46n9kF+4cWameZKW+kZ5ZjL41lgcRe2OGTIa7+Jw1xyaNky9zGnkoj+27OOGsH05OnG6KJz7TO1po2n8nFBE3rzNDYMHJrwlEYBksjgBoCwyycBZiA29mzVudinZXvi2ZQ3hMDgJmdaw1/xqUjvVoyQx4mgiDcM1Y4//ULiqEHyFDeExWcxOWAEH423i0FI7baR6QXouZoyvmDyanCSu+/gyCxg8hRWJDAyLOMMfe12LkuM/EHxZvmL+Fh+AiUIbFcYykPkwjCBL1HxZ3Xf3t5Al5+BCm0iMkxhutjjFokfX9WEdp2soWXbypHIFLA2chjLjw1Z/Gcvity9XCw0zEeLdI6fNwbN7WJa+lEhdJIS84JGsxtAvZIqMsxDRXxkW/3sYkYsqfdhKZkiZkLTJMG0f7ZV6IRw6w2bV5dSJm7vMf5shzsgB2kW9cacGMX+Ra/ZADpzu0vuYCojksX/40bE5PEK+23uQ0swBa5tDOwYMsoTVz7Izsy2Myx2TBvOznzY85CEWM9GSMjblNtncfYM5/kNNGZmO0kEwzkOtB8Vo/GwSKBu0K7FzhZ77Cz5rUppns3roKZ/Xsa94jHS27vonnqa5Mvqwl5kq7xkQ6JFtf2Vx4+Y2WVB+RhRFnplfElqbYpsFLw+T9TPNq0xmHTAqYsXnnB+acfqDssX7GvHAKygYbbk4RnTC0efXvrYfGlzkmz3BOHZjAOUG1LkQ640RfI8ARV1pmfOR1KiYCqZlfF4z8OnYiZB3Z2p3TxOrsmTdlcEoCiuBCgBEEE9Ukt4vyQkHstgelJjQIULaiBAeNoSOzQXHH1Y+wM/EIVBpsE9nCYDEgkOCv7ibeBRdcUGhDCSxHdu6RWwseTQShQ4hauGhZLDQILbIrr1pXa3YdFTk+tBA6miQECUDl6muCGyFGDgkNfWInK766Eij6S9mONWmdvKDFDk9ZNDKO6u38aYG0RZ7aYVHSTlpVdSLM7OYtXOoljp084ktzw2ygHQMIvjKMEfUjEAnsOOKUp3LhZjFWZ2ODBoDAZypiLLPPshjzs6u3iUIkkA+CGxGBJ6IMF/Vyj7S7eqaxUbb2WsBp9mw+2OnCVb9465ldm7jSIGswMbc8953NngVbvuriBToLmQWdFplQD4ew99PP9TOMbESU3TrH5GwxYTvXZU6VH7LC/tCYcd8/rp4q/UKF0+SbM+Y4pYE5oy7GMuJlTtH0ml/GExtd81eccHCniJCPI1rkzRwhBx3F2yjKhwywkaPNMpYifcham8Dwi6vPgRlnZJJ0tdbivQIkgnyyuXMlE/Q72/RI60oOMG0wx9p6I+ZMh8xFZMFcFN8m1/xAJMw/9edP9pD/7sOZYzZ9NN80yeFvjjqloUH0wnAQLTjYtFKI+K6reksDC/VsiTyzKwTJemJTj8SQV+QHGWseSssZa2QbG3/t4hfOJlddEE71Df+4WlPJQn1AboW/K5M32mv9yrab/OFPVsCMNlL9+IUjL2HJkYnh70qWaTv5bHMUpJD8Mm9sIBB44weZNH7gZSzpD/VTNi02LOTZdzZR4jj+F78Np02HAbzgDxPh8pJniz9/jjy3zmiXMcyvddYsbYp6R5gxYJxqmw1IjAHmDGzTYYocW0+ksTbgFu45axria1xTMPALx87beFZvJn/aq37WCGVaqyJurbWYm8aTMRJtjvC8rhsCC0Z+2TmZkCaiHaEdDQEzqPoGuUVfx2+33XbFDozwIXzb+AYj4SIOZwIyPne0xh95MBHFQ2LlR2B6e9/u3jGbdI5yvLBFkCF9CDR/u3cT2KJMgLZlj+u9yeXlMTtkE953HQkHgtLRkRcAkSEaHwsWQUMLwxaOKQPhTENkQuoHx5yEAa2v3acJSqAgw9F/FlAkTBiBiwgyNdFvXjhTjh2vxZXgkh9Bpx+QV/koGwEhVEx2gt/CqZ6IOuGHfHPaaHEnXIwp9dc+xIV2QH9bCPV39BOBQxDRIoWf+lkI1M8ChTirL8KLMCCKdujiEcbaR1git/JWHuFFa6DuFkv1oQVD/i2qdu0EGjtGYwxh8eKaxZhgRWIt8ASpEw/zw+Jo7FoMLeZIC3/YWfS9DARD9tjy0WZzCkmPtsVVWrbDyAABSnibi7Wu3rhEvFFdYU2TDQv11YcWWuOVcGdbbuMxqvooB7GyoNOOKlv/xIInfFxdrbV4H4JmUB2Ne2TOJ7ZsMHx1BBG2MTRntAsREZdDxshF412/iC/cuDVeEA39YU4b++QK0kRGkhfSkSc2XuYpOSrfcMaqMe1kiUwQbtxKiwzD3BwwHpVD9rAJRQpsSo15BIYmzJyIfJE9hJ+MJju0gYxQb3PS0bi6ikMOmTf6k4bcJlh6xBexPOCAA9Z4z8A8NE9sGmg6yTl+tdZik0teInTqJG/1RtzlR1MedSQfmJWQL8q2fpFJNiFkhT6xFiHJxj95ZhMf6V2dxFAUIMZwbsOtX+YMuazPKCmkCQcLm1wbALKLfLDZ0A9eJNa/tf5PBpDPZK/1gnwnwyKvuFovzV3rA5mr/bDVfuQNQRZX2UwIrSHajQjahJNFNvrWEeuyuOGMBdhoj3WLvIywuKq38o0P40T5+ln74YxwR1xX5egnm3vrE14Q40gbyXqbOuOnXd+tA7jEdqu4h7aIJ9ypHbM148sppf5WB9hTIPkRDVpdGwNy2UbA+tbyFnW1rlqL9bv8jFMKEWuN8aDuHNlI8SGefjHv2nBx0k0fgQUjvxZogwMB8pHtybRMhIt4rTOgCNK2qSZLG8c9AWeSIFaeOUQLkZXWQoeY8G+dBcQCYaC3/u4JJoNb+nF3hIaJh1CZvOprQdQ+bbHbNSERO8+cRU88ggpWFiYTMjQDhJo0hBKtLqFLINe6WnjCzTc0aUCQWIsZ4iaO/MMRvnHvSnhYIOWrv5BowoDAirKVi9gSijY20nHih+bQQkDI0xxpg505QUf4aBdnE0NoItSeuX79lGHhkz9HG+6FFvfhkOe4d0Ve5Un4w4YWxIKA8CqD9lpf6BPx4UTzjEzAi58xy85SfHbVntl61VoLbBEL6ZFYbSUoxbXASB+OcObfOkSZsNbvhLwNQBu+EPdOfOAWZau3r1HoX4sU0hBho7zauFjUES/zCOkZZfkzKYs8szBa/I0DG13z0wup8iO3yC9hiIPFmT9nYUfahHH9cOTPZsncEc4hwgimPjS/aHX5czao8g3Hr3VtOCIl71gTxNMGpMiYF86PQ3SQycjXxtAmWBgnHFmNcGNc3uaLcOMfiaBQsYa06b2sZGMcac1D8086DuHkJ7zWWsgo8hKJEW5eIqZIsHktXjj4GNNkFdkgPhlFmWL86zvznT9HbkRa1+g3YVwbjjzzC0duSdO6WmsxNvQZma5/jQ0nm+RzGxfRjrzi2obHPVnnxWeab/EQOTLJeIg4cdVnTiSs3eLCwGYe8bZpiXiuNp7ihLO28+876wIiH/1PhiKE5q6x3sZH/iM/V2uEOolD28uPo0Ro08Is1st+ONzIZrJUGEchgiybc9plDJO3wvShMadMzvzjH878QWo96yNKO/E465vNlzCuHy5OuukjsGDkd/pVXJiYi71UO1OaGFrKUbaFZsJuvt3djrL8ycqi9aIZoGlCsGkeJoufYQuLAHtNRNhGeaFqYuFyCkBrRUNH07dQdclyE4FEIBFIBOYGgZGSX7tRRx6qbifnuMR9urlFgFaDNsERjuMlz3NbwuDcaBH0qaMkmkxaz8ExF8bXLt1xqKNsx520BgtTkyx1MgRsoGj4aWCcBNDATxZ/NmHmBu3MMMdMhYbNmGH76JQo7CVnU+4iSJtVTAQSgURgySIwUvLr2IH9LLstV8dQSxbZBWwYWy12XXCmgfU8iuo4fmK3yXYQ8XZ0PYpyp1sGe2UvAXkZoT1ynW76jDf/CDhCdMzoeJl5iOPPMBmZj9IdQRoLkzlH0jTQyq+1FmPIfbpEIBFIBBKBxYnA1OR3cbYra50IJAKLEAF2gMwMmOywtbWZms9m+IKLD8gPc2z12Puy32Qyg5DTBM9nnTLvRCARSAQSgflFIMnv/OKbuScCicA6IODtfy8LeYnKi4pMH/6fvXsBurWa/wC+nmMGYwbH/a7jWu7JbdyP+y061IgKp4SSS4wUoQvKRBSRIpJ7iqjRSFTkUkREKHQhl2bQDLk0Gv/zeTrr/J/22Xu/+z3n3fvd+z3fM2e9z7PXfX3XetbzXb/1W79nHskXPKoDUKwlUOOhIuHg1YIXkgyDQBAIAkFgogiE/E4U7hQWBILAMASYyfLBDafCqRuwmuGk9bA04wpjbYDKEHN0Tmkz0TWuspJvEAgCQWAAAvEeAwIhv2MAdbGzZDCbziTy4AMPg+rD9BlzX4PC4x8Exo0A25VUHZifoqPuIKwDb/Sz2U9m3sfhTab2WBEZd33kzyoI83pMWFF1UDbzfsLigkAQCAJBYPYRCPmd/T5crwXsRbIpORdZYFfX18HWyyAe04nAEqyVLzGxN8rWKFusvlBVrSlQMWAzlbkxX8pjiH4SEDBrxszaXnvtVdgg79r8nET5KSMIBIEgEATGi0DI73jxXZTcnY5niJ1pr2EVYH2DfuWwOAkLAuNEgLk5JNNX8arrWglhBL7607sdZ11q3j6+wmi9cod9fKfGzzUIBIHJIJBSgsBCIRDyu1BITlk+JL9TVqVUJwgEz2+MWQAAEABJREFUgSAQBIJAEAgCi45AyO+id8H4K2DL2McnSNl8e9wJdp9H9NloOr/s8tbfvqnu88E+Sewb6D4BqYbs9vrsou/R+xyobWrxhMmflIx9Vul8Oe2cc84RVHwcgBSN1I4tVVvbc6ljtAmH/klgEAgCQSAIBIEgEAQ2DIGQ3w3DbWZSOUTkBP3KlSvLEUccUXzWd9ttty0rVqwoZ599dtsO5Lf+/utf/1qQZLqWDswdeOCBbZyLL7647L333gWR9ZGIu9/97uVjH/tYG+ZQnXCn8pmmcpCuaZo27LDDDitItK+9iY94H3fcccVHTtoI+RMEgkAQCALzQyCxg0AQ2CgEQn43Cr7pT3zeeecVlh+e9axnFQb9b37zmxcH4ugyVtWI7m+He0h3t9hii8LE01VXXdU28q53vWtBXOkSn3zyycXhIyf1BcqLNBexlueuu+5apL/sssvKscceW5zgR7gPOOCA8re//a2cf/75xYEiaeOCQBAIAkEgCASBIDBJBEJ+J4n2wpc1Z45nnnlmOeaYY8qVV15Zli2bu7u7h+SYmaoFIM03utGNyjbbbFMQYtJdpFn4/e9//3LQQQeVk046qVCJOOWUU1qC/ac//alIx0YqiTCHJCPEy5cvlzQuCASBIBAEgkAQCAITRWBuNjTR6qSwhUZgt912K9tvv31529veVi688MINzv7UU08tq1ataqXIe+yxR+naPb366qvL5ptvXk488cRWFeLII48sxx9/fGGr9ZprrilXXHFFQaR9vAABv+CCCwpVjA2uTBIGgSAQBFoE8icIBIEgMH8EQn7nj9lMpEA4r7322vLf//63vOY1rym3uc1tyi677FLYVKVv+89//rPQB/ZBDL+R1O5vjWRzVdhVV11VfvrTn/IqdIJ/8YtfFCoNJLuXX355oQNMteIvf/lLIREm/RWPXjCpML1hZPjcc88tX/7ylwvbriTCbYb5EwSCQBAIAkEgCASBCSKwZMjvBDGb+qKQzLe//e0FwSXxPf3004tDbT/4wQ/KzjvvXPbdd9/i61Wkr9/4xjcKSw1vfetbW1Jaf7/5zW8un/jEJ1prDW95y1uKDw4g0K94xSvKaaedVu50pzsV5BcRRrC/8IUvtBLmHXfcsSXVT3nKU1o1C1/HcuBN/iw9kBI/7GEPm3oMU8EgEASCQBAIAkFgaSIQ8rsE+3WrrbYq9G4dLnMIzReyjj322EKC+8Mf/rDsv//+5dBDDy0ktb5mhQjvt99+hcmy+hvh/dnPftYS3EMOOaTV9f3JT35SvvnNb5bdd9+9JdC//vWv20NxDtOdcMIJhemzz3zmM+Xwww8vJL5N07RXVh4ccmMNYvXq1eWmN73pEkQ9TQoCi4pACg8CQSAIBIEREQj5HRGoWYrmIBr9WiTT1UG1m9zkJi3p5Oeec8/Rxx32W5g8fTlOfje84Q2Lg3Hu+XPiyIuf8pqmaSFrmqb4zV96cduA/AkCQSAIBIEgEASCwIIgML9Mls0vemIHgSAQBIJAEAgCQSAIBIHZRSDkd3b7LjUPAkGgDwLxCgJBIAgEgSAwDIGQ32HoJCwIBIEgEASCQBAIArODQGo6AgIhvyOAlChBIAgEgSAQBIJAEAgCSwOBkN+l0Y9pRRBYH4H4BIEgEASCQBAIAushEPK7HiTxCAJBIAgEgSAQBGYdgdQ/CAxCIOR3EDLxDwJBIAjMMALf+973yic/+cmhLfCFx49+9KPloQ99aFm+fHnxdcYPfehD5fvf/37ruokvueSS8upXv7qsWLGiiLvddtu1dr+/853vlCuvvLK8973vbf2F9XO3vvWt2zx9KKdfePXbdtttyx//+Md1RbM3zha58Dvc4Q5Fuep3xBFHrIvjxod8TjrppPK0pz2t3OpWtyq+MOkjP+Jy4vS6H//4x4Vtch/q6Q0766yzhrbHR398sbKm80VN9tQf97jHteWrwz777FPOOeecct5557XRXB/4wAcOzfcDH/hAG9cfX9g8+eSTy3Oe85w2zR3veMfy4he/uM2vW7a4nK9y+mjRAx7wgDb+fe5zn/Ke97ynXHjhhcVHjsTpdT6G9LrXva7Xe73f+sTXQu91r3u1eT/84Q8v2vvtb3+7tSG/XoI1Hr706WNJf/7zn9f8GvzfF0SNLR9P0sfPf/7z27Gi33xM6fWvf31bpjEwyHVzN64//OEPt+NZfF8e9Zu9efbqu3H5aZd4yt9+++3Lj370o3L00Ue30d7whjcMLfsRj3hE+f3vf9/G9ecPf/hD8ZGp2s/64rDDDmtt5hsL4vRzPkD1j3/8o1/QOj9YvPOd7yz3ve992zqxp/++972v/PKXv2zHxLqIuZkTgWVzxkiEIDDzCKQBQWDTQgBpQhiREx+z6df6yy+/vCAZPoTz/ve/vyUw3/rWt1p74Lvuums5++yz22T/+te/WpKDIN/2trctlex4CSOIe+65Z/uBnGXLlhVEzcd0OF+QfOUrX9nmixyoD+LxqEc9qiU24vhoDpL41a9+tY33q1/9qv0wjrTa4AM5L3rRi4pyf/7zn7dfnFTu5z73ueJDPPxUEvnWFkTvVa96VbniiiuKj/A88pGPbOOJL17XIasIJFL0u9/9rhvU3iPTiCjipq6+TPnoRz+6raffyHctH/l4xjOe0RLpAw88sC0fEfWlyze96U3l05/+dJsnG+mIHIKkX3xJ8zGPeUxLXnyU6POf/3xRLyRIGYgXMosYXXbZZYXj9453vKOsXr26zbP+sdh57nOfWyxSjjvuuLaeCO/tb3/78rznPa9YpNS43auveloAwKLrX++vueaatl3ILnvtxou6n3rqqW0ZyKJ61fjdq8XFAQccUHxcqetf740tGOu7FStWtAQdyYYhLIwZfXmLW9xi3bhDTI0ZBBNGl156adlhhx3aMQU7ZVmAfP3rX28JrLGhbbe73e3a8V5x+Pe//10s9HbZZZfiK6S+ViovH4GyIHTld4973KPIS1nIOOKp7/0WH/n9z3/+035F1fPgo1LqDFNxPC9ItX423mvbu9czzjij+ErqF7/4xfK///2vG7TuXt76EQH+7Gc/22Kvz2Gz9dZbt4usdZFzMycCIb9zQpQIQSAIBIHZQgBxvcENblBITb1Ye2vva44vfOELi5c2qRRCKo4P1SCbXu7uEUBfZkTgEBKfKb/LXe4iatl8880LIobg+lDOypUry2Mf+9g2rPcP0kQKSlL11Kc+tdz73vfujdL+RlBINhE2RBzRfclLXlJIJhEIH8lR7rve9a7WTyJEee+99y7if+QjHynPfvazy41vfOP28+oIKcmYcsXtOlJJxOS3v/1tOfPMM7tB7T0JJCLjIz2tR88fEmhft0TW4IDoHHnkkWXlGhxq+U960pNayesWW2zRpt5ss83KM5/5zOKjQK1H50/TNC1+pLwIkDYimojuNttsU25+85u3HxciUSSVhGNNjugjcT5Dr59I8IXBfaeddirvXSOV15/8ug7JJnU1VpBJWHbDkcmvfe1rBWHXz/vtt19BBsVHQHfbbbfyxje+sdzsZjfrJmvvkWbk2xg7/PDDW7/uH2NL+4w/EnpEl2RbHHgh/cq0YNAPJKjCeh2JrUWE/kJGjVVk1q6HBYv0xhUMlWVHQB5Iv90KbSB1lt7Hm0jLDz300LLHHnu0CxHjCebS9Dpli2e8KtsYh/0hhxzSLuLEF0f9LUjUg1/XWQAgysj/8ccf3+6idMPdX3TRRUU9jWN98KAHPYh3u1C1CNIOH5FqPfNnJARCfkeCKZGCQBAIArOBAEJD0njMMce0Eq3TTjut2Abu1v7EE08spEYkSXe+8527Qe09AkeahCCSRiFeO++8cxvW/YNQUVNATLfccstu0Hr3TdMUklMkg5R4vQhrPZAr5SExyBs1BgRmbXB7QS4Rzrve9a5tO7QRmUdc2gidP0gT4tPxam+REcRGnZDJ1rPzBwFTTsfrercIIBJCam2xgZSo+/UirfkhTi0fSSTFXuPd9z9SjJyRJJNK1vr14kXl4t3vfve6PJDLy9ZIhnffffdWSr4uYO0NCbSFwNqf6y4k1wifxYX7008/fV2YG/1PymiMvPa1r22/1sm/OvhYrOj/6levpOkkoFQKqHsYbzXMFSn+1Kc+VRDexz/+8byu5/S58YnI6ovrBfb8UD+LMtJRkl1lwrIbDYZPeMIT2gUGf4SbKg4/iyp+1VnM7bnnnq2Kj7wHLYDEtxhDPI899thCek8KrZ+FdR0Jsf7s+rn/7ne/W9TdwhJOJNf8u05f24F4+ctfXtS5G+Ze33YXQ/zihiMQ8jscn6UWmvYEgSCwxBFAxJARupm2k235kgx2m02a2zRN6Uc6ajx52GK94IIL2ni9ZKLG86JHVOrvhbiSxiIBJGrIRb88EQ7kWzwqBkh4v3j8kEXX6khrqXqQKpPOIpv0hWv4qNe///3vBXmxuHj605/eNxmS3E/i1zfyWk/qI8ghwoScrvW+3gXx56GPzjzzzILAkXTy63XyQDK7/tRKjBWEndQYwbPd35X+IrDnn39+sTMwqP9vectbln79/8EPfrBQP0Bgbc2T7HbL1z7SV5JS0tFuWL1HQi2A6u9hV+0xFhBJC4h+ceVn3HgeqGQY49rfL654o5YtPfUZGK9YscLP9Zxx0DsOr7766mIhirgah4itvjeuagZUZOze6J+tttqqel/vqn8tKq/nmR9DEQj5HQpPAoNAEAgCs4OAw0skbCvXbL03TdMe/iJpJL3ttsIWvd9eyK6DHEIhz7niDUq/of6k17b+m6Zp1RfKkH8IhHoif0OirQ0qrW4mKR1dVKSMhJt0kboAPdB1EUe4cVBuUBrb2fe73/1alQDkkM40Uj9Ctq0EsV88ZJQEnmScJBPZPeWUU4r294tPEkrFBHFCXpF9OrDiklIi/VRVqDDAg46rQ1mwFwdB1f/ue91RRx3V7izA3SKFJL7GIYVWNhUPizAqItQrkLgaRxkWDdpR/TbmSo3CuBllrCL4ylc2tzHl1rQWYPW+e6Vv/pCHPKQ0TdOOBRJ07RaHtB1ZJ9nWp6ToFqb08YVz8B/Uv9RG4Iv8Iuskw1SapIsbjkDI73B8EhoEgkAQmBkEkBkHgpAM+o1f+tKXWvJoe5kEqTYE6XOPoLkOckiblyqCOSjOOPxJdOWLXCJg7gc50jkvf8RnUJyuP2LmcBE8YETPkm4r/WgqDN24c90r19a/ePQyXasjZaRGYFuddJFO8iCJZE1Tr6SpyFyXBAlDlJBOusbKVX9b6Qif+A4Wilcdyb62knAi39Q7LIaEU4shAaVvKh8qCsg1DGp/V9L8m9/8RpLrOUSLtBNxRnwdNhQBUSNVR4jpYMvbYssiQVk1b6QT4dO/6i7txjj5GTfG9Fz5KVdZ6ko32f3GOlJdEttKbGt+L3jBC4rDgX4bCwcddFCrv61c+tSk1R//+McLSfnFF1/cHh6lryw+53F6B1UAABAASURBVPnTv54FY5ZfdXYbHAzUpw7UOTxIelzDcx2MwCZLfgdDkpAgEASCwOwhQJLlNDkpku1R268OyFAHID3ycq2t4ie+A1/Vr9/VNi6dVdvjXuz94ozDj56lrWBSSgRtWBksOiBa1DuGxROGQFBvePKTn9yai4IRRxqH6NED7iUv0g1ypKnMaCFTiGVvPHkiL7b1xe0NH/QboUXILWJYVujGsyBBOJumKRYxyA5ShUwhrt247qmlIOniqQ8/FiEQLARW+zkqFsaFXYJKsqhWkF4bJyxxSFtd0zTtgSskX32qv0WEuC972ctanVl5I2kIuwUAix/iqgvJMInwfDCXtp/TRuoZ8mLarV+c6qdd1Ayov1x66aXVe6OuL33pS4tFAjLbS76RU5nDynhwT12FSkndeYDTE5/4xFYnmZUTCxHxkGp19QzTbefXdfqX9F0fwLQblvvBCIT8DsYmIWsR8OKzqjRhrvVakEs9BNLNzAvWSXOr5Soh6IYvxr1tJOZ8HDpY6PLpUzqcQdeud8JUFimG7dg999xz4NameHHjQQC54EjbxlPCwuXqcBLy6/T9qlWryqq1zmGmLbfcsrXDW5/hHXbYoSC1CBBpcW8tvJhJjZE2B8kQK9vivfFIzhwwQlJ7wzbmNyLjxDxppANXXvy9+TE7ZZueXik1D6oM6tIbz/Y+CRt/+akr0lfxcV29enVrPQJxMd+JO4ojbaRXXQ/N2eIeJd1ccSxeDj744EIn1twA/0FpSAUd3rJQMXfCZVDc6k9CyEKCeVb7OcSXmThxYO6KULFmgGCJAzv+g5x6StuLr7SsNyCbtvVJgUmu4Y4wkk735mk+RA6F94b1+60vVq5cWSxGHD4kbe6Np/9ZoODPuoW8TzjhhNbqCb+ug2OvNL8b3ntPp5mKCcsLxmVveO9vOw4OIlJFgQ/nPWMhZmFaJemILYsfFi/6zc5Ob175PX8EQn7nj9kml8LEZ7vNQ7hQjafr5IUsPxOmLUsPPMJr68ek47fwxXYmaYcJSHcWui7IPkmJF3S/9nqxkRp4CTVNs9DFL3p+dCBJhIyxpmmKMVGdg050Gm1TL1ZFEUNSOAsUW5Ne2otVl2Hlemac9EcYSL5IDWt8z5Sxq+6IMKlY0zTtByoQJuTGNjWJnBcryShSTArVNE1rmovJMZJVL2oHdMRDInzAgK6j7eZaHuJMCmo800kUT9k1vF4RD5JKY4BaBglgDXNlXko+yBJTW0iKOHRSkTxzBOsOnpFqGUE/IQoOoWkPSZk6rlixotDzFI9/rzQVZlQNkCPkRRnqwNVFBRUGdYZPl4zD1tgwTpS91157tTZ1ETdSVISRJFte1VnUwtDhJoSbHVk41nBEDpliy5gUECGydc4agDy1XZvMTdKYIxBl5VgoI5raCQf9RVqsr6kYSI+gkkDCRHrO2NEuz6KFPoshwhFKCwtxEH1kXJ1JpfUfaeby5ctbs2AWTA7fSScvaaqDo3jao7+Fk1gjodQjmPKCrXojnsrx/GmTPIwnFg+0R//Jw6E1WArnqHcwU6ZPSJvlYUFkzFuYGMPys7hit5iVDCSU2gZcjC868/xJhJFZ+XKIuz4g1YaNeRuewjhztN0V+bP4YIzKi+6z9tmd8B7xvlOWeuoX7z7pOWHGMwcnDpYOTHqWSIEtUBBjOGmbvPSlvOURNxoCy8po8RJrE0bAS8hkiQAvFAy2v0yk8vMQe7F4YZsgRtWLk3YSjk4hvTXSjIUuz7a0AwuD8jU5eikwg7OQi49B5U3aX18jNYzzN01TEIbqvFz5T7pO3fK8dEg8SWSciPdS9OLtxpmGey935IbKA2JCIlvrhSB4ISNQCJMXvHASbYSAvxc7AudFilAh0ghEzcNBHC9fW+Ne6NQDbIUjEBYvCFCNi6Cpj3xJZUld+0nhEFO7KuIZ50hqzcO1aZpCGmbu2XLLLduPbiBh5557bmt9Qp3E4zyj9EmRAiQOoREXsSC5tpBCnJAVhAYplK46pBbZURem0brhiKa6CkOokDv1rmldkU/lIz/uETTlUx9gQ9b8IV51iCAC5SAU6SvMCARquKvnHZHWfvMEHC1K9J+2K8PzIy5Hr5j+p7nCYkN/igMP5dtBstimF2w8I07dhSWVEHnbYveRE+OmhlMT0W5zEZwQz6985SutmTILFKa2pKejTPXAGET61as6fcLyg7IRR+FIKPUI9YS7MtTbHECCC5u6la8d3hUWKCTG1F30S+/zaEFkHFsEqD+psrKNdzasK0lEMNVFP3huvJOMQe3WXmXXuruqk/zkYTxJ1x3XTdMU/an+FkH6WH9YECD6xoO+QWbt+CnbTkxVb1CG/EiNlU2NQrj+EqZd8mMCUD8qRzubpmn1hS2UxIsbDYFIfkfDKbEWEAEPv5eQE9YeYi9gL58FLGJmsiLh8QKYmQqPoaKIE4lNb9bInE/hknb0hi3k77nyMk4dUiGJIwH10QQvqbnSTTKcChGCw3lxIrC1fMSEf3XdcBJbH1WwxS7cSxvBQyZr+npFrnx0AMEUlzTWwrh3/CLDwqvzUraorfnUK4Jd47iShNWwekVQEBOkSxxSWWQSUapx6tWzRJJIBUBc/YRsWcCIY5HJnyMB51edw2D8q+uGI2E+OFHDXOmM1rT1qnwHzBBW9RRPXfRN7/j2W3jXkbDWvOrVooAKBOl8jbv//vuXHXfcscClxqtXCxb42/YXH6bUYBBakm1b56TjwvSj/qtplaXuwrjecDgaO8aIcPgaD6xFGG9Iajf9ijXS9pq3qzRd1w1H7Lpt1N/duklvLNAD7+bh4yYWdsK7bvkaSbS6kiqLDzP2oo33bjwLDDrNtWx4aRO/bjz3vc+RZwYmwroOxnYB9L2yOQsn5dc+qP2jft2y5GfsScMJ90W5mj8CL28LcuHqC3PjXliNl+vcCIT8zo3RTMawmrRF6WHyYFhx1u0hW0mkrh5SOke2mDSSyRxmdOjamXy87Bw2scqn8+uFbwXb/e0BJ30wudiykQ+pAymICc0k7+VnFSyMs7KVt8l23333LbaH1I/EhORBHI4qAGmbSdKKnGTY9qQHnjSHdIHeoodfvWyvkgZY2ZPS2n6VD2fV7nAFImUCJVUkSbAlTJpCZ4+USF21x6RDkqFM+cJSufzqby8425WkX15cpDzK4kg1vNxNdqRl4tm2EzbMkahJw5F62hLWPjiR1nV/w0pbSMVILWr+pEi2rOVBEkGqIq1ySY+oEugbOn4kIvw5OJiIkT2Tq7bzXyxnN8BW4nwOCo2rrl6wCLA+9XwYs+MqK/kGgSAQBCaIwCZZVMjvEu12L2oEx5YJ6RnygxzZtnF4hzQAsbUFiFzZTqLWQApryxJhQyytjOmiIazIUNM0pf62jWNbk76UPG0XiSMPW2PytC1Dj4vOE+KIcNpiRULljWySNLkijHW1TyKMmCHjtpmQbESVxBiptfWJ/ApHdhElbVYWIqgNVvMIue1ZdUQgbctZDCDPttiapim2Q7XPNhISa+WN3Ki/+sLONhWCXX8jnsqHL/069bIdZThpH4kEwiycJMbWHn/hgxzsbKdpL1062FlIWMjUtH5rgwULAo+kIr+kEOopHqzonNkORPj1DX/lCiMxgAmJnXorlx9ibStUX1ngkOxIM0kHV07b6UROqmzbsCRLTdO09jibZv0ryRbiC2eLNZhNqn4pJwgEgSAQBBYOgZDfhcNyqnKy7YM0VfLl2+LIJsKEOCE/pIwIEBKHUCG7TdO0JoCQSgSE1JWUVjwNtEVdf5Mc0o1CsEhLkS2OdBHZYgMSEVUXh1YQNyoPiC3CKr9BTnmkt7bUkGPbaogn6TBpctM07dYfvS7SOJJmkk46UQ4lkCojwvTo+JGAktCqP10q26YIN6miuisPRtqDrJJ+0uvjv9lmmxXx1FW+fjtgIA6ckGlbo0i5OAgkvTS6kQ5ckHojzvAU3s/BxOEGOl5OYcuvaZpie5X0s6axQPEbfrbnSLpJeZFYixvts41t8WIBYCsWWbPwkId6W/QgevQTbZ2qqwXLsmXLijGB9Etnq1eaSTq2Rznkv9/W+7jqAlPb6baEhzljSB1Iz41l93FBIAgEgSAwWwiE/M5Wf41cW1JP0lVSTNIqBBSRI+F0j8DZyiW5pbhPr0zmSNGopKNLCOUtPdUKJBA5kz+iTDeQWgQCS5UBqaTPKf4gJ27TNG2w+5p/67HmjxPJTXNdOFUIZFueyuSciHXQQVssAOhDNc118RFo0lLS4zVZrftf20PCZ6GwLmDAjXrVoG59lIeoqwcHYwsBxLrG771SRSE5hg1Vld7w3t9IePVDeOtvfYsMOyTjgFLTXNfmGpeuGVNOFhYk39QcSL5haBdAfZF6kmQHWWq6SV0dKONI3pH4SZVrvJLkKnuQoypkkWJhIg4p+qTql3KCQBAYPwIpYdNBIOR3ifY1EkOCyC4gdQKnd724ETakmDTVYRCOxHehYGiapt02doBE3tUhlggh3dsqaV2oMhF2Tr61PFd1oOah3EpIa5lIMaJYfy/kFVF3sAJ5U4/qnBofVA4yTqeYCgXd60Hx5vIn7d9nn33Kgx/84EIy7JBFNw0ijuQ5LW1RQrdZfeGnDrWurvy6aSd5b5waw5Msc1hZdkoceKEigviSrA+Ln7AgEASCQBCYXgRCfqe3bzaqZvQ8SWBJQ0lBSWTpv9omJ/Gj3+qAli1v+o70aYXbOhcXYVQBv22Zu1Jb4Ces+xsh4Gdr32ljJNSpeLqbiLf81UVa0lZ1cM/ZXidlVb4DdbaS5dfdqld+zV+5wm3zq688kHlqDMgclQtlIYFUDpy+dliPCgSVAmmVQSWANFp6eXK1fddee22p5Ql3Lxwufrvnanzh6qv98qdrTUeYrUykW32Y87rnPe8p+fWc+PKVHx1hW+50kx2Q027+4tS2Ssyv/navbHmIryy4k277TdIurjaLi+zKw+KH5JIUmGoF9RaE1+KEJJi6CB1gccfttK9i6dDhuMubT/6wZarI6XqYWphQfTBu55NP4gaBIBAEgsD0ILDo5Jd+IePcvZAgQsgLyWXXeSmLa/veVnINQ3rol9bfg670Kr3c5bGUHX1T5Ia5GpYSqDWQprHByEbkkUceWVyZrUE8bNuzXoBEedlLj9TZhqeripQgc7bn2XesvxmDp1OLWMKWpQFSRZJeOrTUL2zBs1MJdxJXJKJiTweV5Fn/sTZgqx7xo4NMwqaf1Z9ag7oqSzjTaOrj8J28kEaWK+htknLTV0XmSDW32267QmrHVBXpKsfskUNh0jM0rz3S0Iem/6p844/+MFuSyDQ9VL8drrNooMfsYJivanHaDgN6tNQKqJPYKkdm5U+tQF27TnvoYSuDAXbSagSWugopMOzlq17C6ROff/75Be50di0sWHNw2NABNYTXAkRa7UDy9aF0yC8s2fs0Ljx3sKEr7QAk6x877bRTsViidwufbl17xOgVAAAQAElEQVTHcU8fWln6X/2ofmjrOMrakDzNI3ZQ7na3uxVj07jZkHwmmcbcaS50cFH96cNbEFFtoZOvLsaMsEGOypB41VmcmhOMb2mE+7CG/vMM1XjmDM+GZ0o8i3DPi3jGaI3XvdqRGnS40XMon0HOM6oeNT/j32+HZaURrt7a7lmwCDS/CRvkCAcseuTZ2x6qQJ4bi0rPoTjViet9Zk6Qt6s2W0RaGMuT/yDnefZc1/w8DxVLH+cQbp6BtzbWeK4WuPrYHCp/c4/6iUsA4cp/mFN3ebVuzR8fc6jjxOFmux8EJvJbE7zuP0xhDBv1tNDXZnHhZI4yDoaVbY6qGcJJ+8xf0ihb/sagfGs8V/OqMGXDR9nONChTuHt5DHLmHu8ucTlCFTh6Z0njGfIsaWNv2fpHvb2PvE+V7x2h3ZWnyDNuOhFYdPKLcNiG7X2gSANvf/vbF5MHYuN0NULFgdLVw4BYSEunk1TGYCXtFMeBJmmRNX6c+CZU4UvZOQjGogFpI1IIP0QTyWX5wMLBtjgCbHteGHuCXhqIlwNA8KH/yuSWg1AOn9EvdSCs/ra9j7zqQwfJ9APJLp1NZJrlAv6kwfJCjOVbHULu0BirBWxR2k5GXr1kEWb9zN9LzNWYYDnB5I4oIrfyolusTBMQ4ol0V8sRykac2VqEx8qVK9tPmSJ90rOb6KXB3Bl8bP8j3sysiUOSi1jC0G8kFoH2G5FHGlnMgAldauRT+X7DmkTX4UBjWl27Tl5IvHFKgk0C6wUCDyRaftQYkFm6wNovT8+Nw28WE0gqSb56ap8XDYzopsIWJltvvXWrjiJvknDx4AEr9aEOAWP64frIGFCesHE6mGgjQu7FpW+MqXGWOZ+89YGx4yUJs/mkXYy4CIvxYVFlUWvOow+OxJgP7LyoF6xhL9yLXRtd/RbfeBePozpkPPgKHKIqDpJpMWCcVcKGDCBnFk+IkmfJPGLhat41D8iv1yEX6izv3jBlW/x5Nox9C0DE2jxDTxtpRkalQz4RfQs5Oy/q6VlERNTJM438q6/5TrgFuWdJer+RKYtaCwjtQWyktUOiPZ5/hNQirT470iK3LLwYx8LNC+Y7RFp6uMBM3albKcuHNDyb5kC/EVTPoPw4aXfbbbdicSyN9nt3KaPu4IiHYFqgwVh7YEVlSV97/vWTOllgw01ZSJp5xT0nX3OD/BBA/U+QYMGgbPMMQYTdNXOlsSUuqz7w9t5FWvWJnRzE3zyi79SD8EA5nm3EUl38Nj70P/OR8oOjd5NyCDjg7R1gXGuLOV48zntfPGUT2sBHHbwbvEMQaOTXWNAf5mnjXtnKNb9ZaBMMyE+7EV5zr3Gnftqu3fq64iMuBxv9Y1fRe0T5FnnehdouTtz0IrCo5BexNSlbaRlIXZg8bF7YiBV/EwnSRQLjt4nCROjhI+G0FWkyN+kjGuyfSiOuNPw4kzy/pe4QOPqe2uxqUmqapm22BxoBIhlFVBFakxISLD7npb9s2bJiwve7OkSl3rsizMi1e46uqYkXGdVf8kQ+bRObgFhqaCux9k/TNAWJExdZ7c2/9zdypJzqtGVtVkX+6qJM9VJmDTPRCdNm1hlMfsKkr3m5SmfsuOeQcdJy91zvb3FhLYzz24uVYxFAeYglfJXX67rtsbjojlX59YZ3yxLeG9+Lj2ULCxaqINrpoB0cm6YpxgKyL6226PtaJ1JNljv0hT6p/uO8Gl/qpD7VOYQ3zjLnkzc81GsWiC+yxqwhMoSQWNyquzGImJkDEFxEAREVVzjVIOO1jjWEFnmAE4KBWCERFlUWddLY0ZCHHSWEUVzzuLTGn4WhhbW4PjaAPCBA4nWdupD6n3XWWcWuRDfMvTqru69haYfFofnEhyPsWljkWySLSxrtQKe6EXwoW5xaDyQc0bRYRE6EW/B615gb/LaINR4RVe8n5MZz0m2PRYT2UGlSrjZYHCBtykfI1EF+cJQe7oQ13lV2V4QZ557POsb0mcOq8kR4tdcCnwqZMqWhfgNPJE88CwZk2A6RxYkFLayUrxxznTjKVj/p5WN+Rtjcc8rw/MsXIZcntTB1ko84FvHmFe1FcF3tppF+igsr4TDnb85RtnlIveRh7qHL73nyW7uUZfzJE3FFvMXXF/rKDqKrPlOWdpPkK8e4hHl3XBr35ntxzY/ygcmqVata6zna6R5GhA7MS8pTuRYf+kwfqp8ykG733fGrXPOk+ctChKBEHH3juUCk5Rk3vQgsKvk12ZEyWHVaEXuY5oLKw0mSKz5Jr5Whh1g6dl5Jv9wPcgYmUjIoPP5BIAgEgVlEwMKexIqkEfGwCNUOi0DSM7sVyIAFjwURf+G9jr+XOTJNqmquRTK96AkYxJe3PBEe86ltdyYHSRYRGou9pmna3Qbzs0UxaaC0XUeNyhxuUYdAIWndcCTJnN40Tdd73b2dEwt5W80IqbiEJlXw0TRN8Y5BduCCTFtka+O6TDo32gcbpBShus1tblMQJwS5aZp17UH6bcdLSqrrnYTwyVt7m6YpyoAD7BBAkkd1bZqm9PsHT9iRHlpwaIM66K+muS6NvBF2Ull5MFNZd+v0rfCmadqy5WfnDUmThuChaa7Lp/T8gxHpu0O5pOeIt0W/OjRN07bbPcECQqht1EeQdL+dGzAmZOuKzJKUGofGh0W5sF7XNE3RZwQEpPJ2Cy0+LBjUqWmuK9u9BYpFhjyQVG33PleGfuOvbGVRg1AHxBbZFdbP6Rf1I522MEFgqcgpT/ymuW78IOAWDvxwEHERcOTZ+IWHMPVQpgWE33HTi8CyxaoavUYSCoNKHWw1WSW6H+SE26KyOrS6MxkPihv/IBAEgsCmgoAXMckfgkNa2K/ddjnskPUL6+eH/JqXEUwEpV8cJJEUkX6pLWQ7J373i4vUdP2pKVAnILklYaRqYEu9G2fUe4IQxNnOC8LZL52wfv79/Eh9qUdoD1LZL478kH7kl4ScZLRfPKoXSFa/sH5+VAAsYhAxhLlfHMSUdJp6CVUBUvt+8eyGkZb3C+v109+k93R/LRZ6w/1GxO0KulIjgU3dnRXedYi4RULXb9g99QeCMGO0Xx/KzyJKP+MAyLiFSr88SX4tRPqF9fNDlrXJwqlfOD/E35XUl6qEtlu08et1dvF6/fJ7uhBYNPJL78e2g9U6Ekunx4AaBI/JnZTBiouuFD2uQXHH4Z88g0AQCALTigCdTtu8CEEvcaAKRshQne1vhGmutiBD9C4RMMRgWPx6aIgEcFi8bpg53WElUknWR5BgdSUY6cYb5Z7EsmmagvyPEn+uOA43Nc11KlnD4sIR8UXkkaFhcUcN05fyHSaxlJcykUVEjzoHv41x8qPKgVRTTZgrLwsE0lXkfq64o4TjAMYuSeqw+FQeqZGQ8mr7sLijhpEik97aueimoe9rR6M+O6Tj+Ae1BovCbtzczxYCi0J+DV4rZlsbVpH0ksBmUnbt5xBkul/CPKC2hWyT+B0XBIJAENiUEaAvqf22ZHvVx+hZ0pF12Ec8EsW5yKy8OETItZQy9DJqvG4mDoTSN3ZQCIGyle/sB2lwN94o9xtS/rB8R83PAgFpHJbXfMP0kXxHSSfuKPFGiaPM+eRnrI2S76hxRs0P3uo6ar6jxNPf8ux9dqjdUJdBjlktId13EE7cUfJNnOlFYFHILzMkBjAFcts7dcXsFKYtl35wNU1TbGPY3kKEbUlRRKcK0S9+/IJAEAgCmwoC5lASMwIBp9y77ab7aWu6aZpCWkUnsmmabpS+903TFNvWDorNNc+S2tqWJjEdhcQQYLBy4vS+eZ1agANHJGoOMtN77VupAZ70NhESkmrvlgHRRvZWH+0h0R7WHrrApN10PakMjFzAkIj6yDtOfsPaQvJJHYXEn8rIkCxHCpIfskeKTAo7VyLWd0hglT9X3FHCqTuQeJP8D2s3vW0SX4RV/4yS91xx6GITyFE5MY5qfGPAwUQSbvfGBYm8xRqMjNcaN9fZQmD+5Hcj22ewWN0zP8J8EMV1D48BLWs6va6DHCmx05vLly8vDmI4KWr7bFD8+AeBIBAE5oMAKY8DKxwyOZ+0ixUXCXOgzVxKdWAusjpKPW0DO8yEWLFv3S+Ncxt28pBrJ92ZtqLK0I+80JVEbpAWu3wOC3kPVIfs8aP7SUDSr7xBfnQ11cE7oR8hQmy8dwal7/Wny0ktz1hwGGpQexBj+rEsULCdjQz25sXPAqLXf9BvOtvImEUAU2698eAHS2SV3jHJOSl6NT/WjU8/FpkcRaLbNE2xiKA/7DB6vwWIMrRFu1nA8HyQiHqvd8t1b9yI634UZxdA2+mZO8DYm0YbCMy0mzUGfWKsq1NvXGOS2bde/0G/7TrgIHSJHcLsEuDeNBYI1CD0gbqqV28cQr2FeAZ7883vhUNg4uSXzUGSCZOfVT/nns6XQU36a0ANaiIpBh0cE62VmBcUszYkCYPSbMr+TMGxO2hFuynjkLZPJwKeW4QH0eg6ZrjY70SuJl1zc4wXmxcie64IZbcO03rPUoC5kXqDOZFOproiIQim+17nxa2tCFo/dQOmEG37srvNmg5igEgiseIjaAiWfIWTFCubP11Y/oiMDxbQn0RY6EyyImELWXh1yDZpovxIgWv6Gq6O8jCnua/+riRxBCnyYO/WITQm2BBF7xsWDHrzUxeEGEEzDuXTdcx/kfTJD6mu6enkOrMCC+U5hMXclgWCuOZaZBCBchCQHVht6uYNP7uXCKb8umGkjCx3wJKghxkzY1B+4rKyQMdZGpJfKoDeq/RRlSdvYYg7U6Keoab5f0k/UubMDGzgKW51dH3lg7wZS2wNV6xJ1Y0tBLRpmuLQnecUUfaOqXWSv7GBSCqj5u2KLGuDHV79w686qjhwFEcdHKCsgi31ZF1DPzVNU1iw0G72zZWtL+Sj7epshxg/4Fed50AdtaO3P0h3jTn9xswaO8jqh1hbiMm35uPKggfrDwcccEBrog8m/GFt7FsE4jP84qYTgYmSX7pnbAayF+neIDe4DDqTo4nY4EFsvXhsL5hIQefht7VmYHpwGB/3YNHRQZ5tnRmwVqROoXow5S+tE8QGt9Wg35uKMyGQrntATWabSrvTztlBgLSF+SIkgJ1Wi1kSTC9+/qtXry5eWpNskRP+dqAcrvVyZYKrEoBJ1mO+ZTnshhB45s2rq1atam1fmzcRQ2oGDhjXfJEp5MpcC3/zBHNSyEmNQ6JszvUFK8RKuENlSCpCwiyUbX/x+TNvZf4mBUXMmKFiyUE/s/aAeCDTiBnpqjlKWs7crc7eAcimOb2GI13eBwgjYiuMRFa66pBQhEv/WVCxcsDslLobT6SpNa4riak+RtboQTMLxr86tq6lJf32kZnaHgsiJJHUlYoAs1jMoZFYw5RNWuHqwYaxA30kwzVfB7t9gAZhRczlo9413JXFDG2Wrw/PeH/BUNm2/O2AiqfPLRoRbATNPcxhK38mymc2FAAACHJJREFUxKhRIOniI2X8kFPxvSe9j4VxCJu8EXl463t1Vx8WEYwlWFLzQC71rfetd7IzPKxL0C+3qLC4gYN8OQse7dZvsIKnd7YwTh1ZSWASTxvYjFauc0HGr7qSTCsXUSb91T/KVme4EKJRvWGZgRqQfDmLH3XXfm1Ttnth1dk9sKhZuXJlYbvX+DEGmP0zzi06alyqQ+Ypz5s06qV/Vq9e3UbRpxbR7Y+l92dJtGii5NeEZqJBWBk1ZybH5IHgehBNHFbj4vjSionBPX/EGAH2oDEETlohrjCTpsmbsXAPhgcW8ZVGuPKsdA3YJdFrIzbCpMFgt4cSDiMmS7QgMDEEvGzpq9L3a5qmeEa9cEiUPM9IEknhxCq0tiCkAun1crN49pKfNAlfW5V5XRAxJA+hseAlrSQtQxjZoUVkaoZN0xR2Vc2x5lJzMKECElvjuMoTKWFPloSLEIFqA3KCcCAt4nHmdGYoETxzsDogM7a05UM1gXqDOZvE1RwlHWfuRnzN2wQbtutrOAJq/udP8ixvY0a6rkPEEVrSOvGRbW1ElhG2blzYkACaG0kY9XM33D1LF8i6MahM7aFiAA/tEYczjhEqO5neWzCyOHBIG4HrYoQMVwy0x7giRZRP15Fms3BEAmoLHx7KRl6RvxpXu5Bj0nmSTWVbqHivInDqVuP6AIkdAfjrc3h71mq4q7oin6TPiGXF0QIAqVaeeBwMVq4hi8ilvGBqrFlkWTg1TSNa6yyYvKeVrd3Gkme8DVz7R9nmA2XZrVC2BRCiCd/esnEB5NNYghFSjawah2uzbC8WJPpPucaPfGHRBq79U8v2zMNcHFgi6xbl+m1t1PZCQm9x4pnRN+pA6ov8a3sbKX+mFoGJkt+pRSEVCwJBYP4IjDkFEuYF7YU15qIGZk8yikSQSpNEI5IDIycgCASBIBAEZgKBkN+Z6KZUMghsOgiQ0JAWkjTaYrWlOK7W25YmvRnkfEHKdi0CTueQJI9t1XHVJ/kGgSAwOwikprOLQMjv7PZdah4ElhwCttHpadJzdJrclibdzHE11LY4kj3MIchV59e2psM046pP8g0CQSAIBIHxIxDyO36MU8KSRyANXCgEnOSnXkB/DwmmO9c0/683uFDl1Hzop9L5HOToJ55xxhmFeSNqGMw6SVPT5xoEgkAQCAKzh0DI7+z1WWocBJYsAg7yOLDi9LaP2DiVvpiNdYDGQTGHj5z2dlp9MeuTsoPAVCKQSgWBGUMg5HfGOizVDQJLGQEWB1gnceLaSXLXXpNEk2o/6zRMZt30pjctTnuPU/d4Um1KOUEgCASBIFBKyO8SHwXMtThA5MqE0Jibm+yDwLwRYGaKaSqqB0wgysDHBZhuYu6ImgGbpcKFTcKxm1rNT7Fd28+s1iTqkTKCQBAIAkFg4REI+V14TKcmR4bTGW5n85DdZC/xqalcKhIE1iDgwJmPIrDbzY4nFQO2vJumKezVHnzwwYUdWgbtGbpfk2Ts/z03PvjAQL+va/lYQdM0Yy83BSwFBNKGIBAEZgGBkN9Z6KUNrCPJlS/tVOdLThuYVZIFgbEg4AMAvrhUx6irj9TUwkh9+XE+qlD9x3n13PgQgDJ9HWucZSXvIBAEgkAQmDwCIb9jwjzZBoEgEASCQBAIAkEgCEwfAiG/09cnqVEQCAJBYNYRSP2DQBAIAlOLQMjv1HZNKhYEgkAQCAJBIAgEgSCw0AiMn/wudI2TXxAIAkEgCASBIBAEgkAQ2EAEQn43ELgkCwJBIAiMgkDiBIEgEASCwHQhEPI7Xf2R2gSBIBAEgkAQCAJBYKkgMJXtCPmdym5JpYJAEAgCQSAIBIEgEATGgUDI7zhQTZ5BIAisj0B8gkAQCAJBIAhMAQIhv1PQCalCEAgCQSAIBIEgsLQRSOumB4GQ3+npi9QkCASBIBAEgkAQCAJBYMwILDj5veiii0pcMMgYGDYGEpbxkTGQMZAxkDGQMbChY+CSSy7ZKHq84OT30EMPLXHBIGMgYyBjIGMgYyBjoO8YCE8IT9rIMXD00UdPF/k96qijSlwwyBjIGMgYyBjIGMgYyBjIGBjHGDj44IOni/xuVG2SeFNDIO0NAkEgCASBIBAEgsBEEVhwtYeJ1j6FBYEgEASCQBCYWQRS8SAQBBYDgZDfxUA9ZQaBIBAEgkAQCAJBIAgsCgIhv4sC+/qFxicIBIEgEASCQBAIAkFg/AiE/I4f45QQBIJAEAgCwxFIaBAIAkFgYgiE/E4M6hQUBIJAEAgCQSAIBIEgsNgITB/5XWxEUn4QCAJBIAgEgSAQBILAkkUg5HfJdm0aFgSCwCwikDoHgSAQBILAeBEI+R0vvsk9CASBIBAEgkAQCAJBYDQEJhIr5HciMKeQIBAEgkAQCAJBIAgEgWlAIOR3GnohdQgCQWB9BOITBIJAEAgCQWAMCIT8jgHUZBkEgkAQCAJBIAgEgY1BIGnHh0DI7/iwTc5BIAgEgSAQBIJAEAgCU4ZAyO+UdUiqEwTWRyA+QSAIBIEgEASCwEIhEPK7UEgmnyAQBIJAEAgCQWDhEUiOQWCBEQj5XWBAk10QCAJBIAgEgSAQBILA9CIQ8ju9fZOarY9AfIJAEAgCQSAIBIEgsFEIhPxuFHxJHASCQBAIAkFgUgiknCAQBBYCgZDfhUAxeQSBIBAEgkAQCAJBIAjMBAIhvzPRTetXMj5BIAgEgSAQBIJAEAgC80cg5Hf+mCVFEAgCQSAILC4CKT0IBIEgsMEIhPxuMHRJGASCQBAIAkEgCASBIDBrCMw++Z01xFPfIBAEgkAQCAJBIAgEgUVDIOR30aBPwUEgCASBjUcgOQSBIBAEgsD8EAj5nR9eiR0EgkAQCAJBIAgEgSAwHQhsUC3+DwAA//8RCUSBAAAABklEQVQDAD2i/1p2RYeEAAAAAElFTkSuQmCC\" width=\"695\" height=\"281\"\u003e\u003c/p\u003e\n\u003ch3\u003eHistopathological Examination\u003c/h3\u003e\n\u003cp\u003eEctopic endometrial and uterine tissues were fixed in 10% formalin for 72 hours and processed using standard paraffin-embedding techniques. Sections measuring 5 \u0026micro;m in thickness were prepared, deparaffinized in xylene, rehydrated through graded ethanol, and rinsed in distilled water. The samples were subjected to a staining process involving hematoxylin for a duration of three minutes, followed by a thorough wash. To differentiate the samples, an acid alcohol solution was employed. Following the application of a bluing solution, the sections were counterstained with eosin for a period of three minutes. The process of dehydration was conducted through the use of graded ethanol, following which the sections were cleared in xylene and mounted with Entellan for microscopic evaluation.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eImmunohistochemical Staining Method\u003c/h2\u003e \u003cp\u003eImmunohistochemical staining was conducted to assess protein expressions of PI3K, p-PI3K, AKT, and p-AKT using specific primary antibodies (FineTest). Antigen retrieval was carried out using a trypsin solution (0.2 g in 200 mL PBS) at 37\u0026deg;C for 30 minutes. Endogenous peroxidase activity was blocked using a 3% hydrogen peroxide-methanol solution. Primary antibodies were incubated overnight at 4\u0026deg;C at appropriate dilutions: PI3K (1:50), p-PI3K (1:50), AKT (1:200), and p-AKT (1:200). Following secondary antibody application and streptavidin-HRP treatment, sections were visualized using 3,3\u0026prime;-diaminobenzidine as the chromogen. Counterstaining with hematoxylin enhanced visual clarity. The staining intensity was evaluated semi-quantitatively using an H-score method, categorizing reactions as strong (+++), moderate (++), weak (+), or absent (-).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eThe data were analysed using SPSS version 25.0 (IBM Corp., Armonk, NY, USA). Continuous variables were presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation, median with interquartile range (25th\u0026ndash;75th percentiles), and minimum\u0026ndash;maximum values, while categorical variables were expressed as frequencies and percentages. The normality of data distribution was assessed using the Shapiro\u0026ndash;Wilk test. In instances where a comparison between groups was required, a One-Way ANOVA followed by a Tukey's post hoc test was employed, provided that the parametric assumptions were met. In instances where the initial assumptions were not met, the Kruskal\u0026ndash;Wallis test was employed, with Mann\u0026ndash;Whitney U tests and Bonferroni correction for post hoc analysis. Paired comparisons were conducted using either the paired samples t-test (parametric) or the Wilcoxon signed-rank test (non-parametric). A p-value of less than 0.05 was considered to be statistically significant in all analyses.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eAnimal Weight Findings\u003c/h2\u003e \u003cp\u003eWhen the initial and final weights of the patients were compared, no significant difference was found between the groups (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eELISA Findings\u003c/h2\u003e \u003cp\u003eSerum estradiol (E2) concentrations were significantly elevated in the Endometriosis\u0026thinsp;+\u0026thinsp;Metformin group compared with all other groups. Despite the absence of statistical significance in the differences observed in progesterone concentrations among the groups, the highest values were recorded in the metformin-treated rats. Levels of vascular endothelial growth factor (VEGF) demonstrated no significant variation between groups; however, the lowest mean value was observed in the control group, whereas the highest was detected in the endometriosis\u0026thinsp;+\u0026thinsp;metformin group (44.53\u0026thinsp;\u0026plusmn;\u0026thinsp;30.2 vs. 86.93\u0026thinsp;\u0026plusmn;\u0026thinsp;42.2) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eStatistical comparison of blood hormone levels of groups\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEndometriosis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eEndometriosis+Dienogest\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eEndometriosis+Metformin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eE2(pg/mL)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA.O\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e S.S\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e91.1\u0026thinsp;\u0026plusmn;\u0026thinsp;52.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e139.77\u0026thinsp;\u0026plusmn;\u0026thinsp;178.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.35\u0026thinsp;\u0026plusmn;\u0026thinsp;19.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e308.65\u0026thinsp;\u0026plusmn;\u0026thinsp;152.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.05*\u003c/p\u003e \u003cp\u003e(kw\u0026thinsp;=\u0026thinsp;7.741)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMed(IQR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e95.83(40.89-141.62)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e95.83(40.89-141.62)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e81.25(54.67\u0026ndash;87.13)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e349.3(164.34-435.91) *\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eProgestron(pg/mL)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA.O\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e S.S\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e113.35\u0026thinsp;\u0026plusmn;\u0026thinsp;12.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e146.71\u0026thinsp;\u0026plusmn;\u0026thinsp;106.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e170.01\u0026thinsp;\u0026plusmn;\u0026thinsp;77.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e199.99\u0026thinsp;\u0026plusmn;\u0026thinsp;35.89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.139(kw\u0026thinsp;=\u0026thinsp;5.487)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMed(IQR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e108.51(101.38-124.15)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e113.55(74.11-222.93)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e167.36(99.8-242.88)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e210.48(195.06-220.03)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eVEGF(pg/mL)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eA.O\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\pm\\:\\)\u003c/span\u003e\u003c/span\u003e S.S\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e44.53\u0026thinsp;\u0026plusmn;\u0026thinsp;30.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e68.75\u0026thinsp;\u0026plusmn;\u0026thinsp;64.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e54.28\u0026thinsp;\u0026plusmn;\u0026thinsp;44.74\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e86.93\u0026thinsp;\u0026plusmn;\u0026thinsp;42.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e0.577(F\u0026thinsp;=\u0026thinsp;0.675)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMed(IQR)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e56.46(12.98\u0026ndash;64.15)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e54.16(18.01-111.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e56(11-95.83)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e70.02(61.01-109.54)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003e*p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 statistically significant difference; A.O: Arithmetic mean; SD: Standard Deviation; Med (IQR): Median (25th \u0026minus;\u0026thinsp;75th Percentiles); kw: Kruskal Wallis Variance Analysis\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eEndometriosis Macroscopy Findings\u003c/h2\u003e \u003cp\u003eOn the 28th day, a comparison of the lesion diameters among the endometriosis groups revealed significant differences. The Endometriosis\u0026thinsp;+\u0026thinsp;Metformin group demonstrated the most minimal lesions (3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.36), which were considerably lower than those observed in the untreated Endometriosis group (177.8\u0026thinsp;\u0026plusmn;\u0026thinsp;214.01) and the Endometriosis\u0026thinsp;+\u0026thinsp;Dienogest group (103\u0026thinsp;\u0026plusmn;\u0026thinsp;143.58) (p\u0026thinsp;=\u0026thinsp;0.002). Despite the observation that lesion size in the Dienogest group was reduced in comparison with the untreated group, this difference did not attain statistical significance (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eRT-PCR Findings\u003c/h2\u003e \u003cp\u003eWhen endometriosis foci were compared, p-PI3K expression showed a statistically significant increase in the Endometriosis+Dienogest group compared to the Endometriosis groups. It was determined that p-PI3K expression increased 2.96 times. When PI3K expression was compared to the Endometriosis group, a statistically significant increase was detected in the Endometriosis+Dienogest group, and the expression of PI3K was found to be 1.88 times higher. When endometriosis foci and the Endometriosis+Metformin group were compared, no statistically significant difference was detected in PI3K, AKT and p-PI3K expression. Although the expression of these antibodies was found to be higher in the Endometriosis+Metformin group (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eChanges in gene expression levels in endometriosis foci of the groups according to RT-PCR results\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGen Symbol\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eFold Regulation (compared with the control group)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eEndometriosis+Dienogest\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eEndometriosis+Metformin\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFold Regulation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ep-value\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eFold Regulation\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ep-value\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBeta-actin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003enan\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003enan\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePI3K\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.002*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ep-PI3K\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.01*\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAKT\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e*p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 statistically significant difference\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eWhen the expressions of the uterine tissues of the groups were compared, p-PI3K expression in the Endometriosis group decreased statistically significantly compared to the control group. PI3K expression was significantly decreased in the uterine tissue of the Endometriosis+Dienogest group compared to the control group. When the protein expressions of the Endometriosis+Metformin group were compared to the control group, it was found that AKT expression increased statistically significantly in the uterine tissue (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e\u003cstrong\u003eTable 4. Changes in gene expression levels in the uterine tissue of control and other groups according to RT-PCR results.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"570\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGen Symbol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" style=\"width: 503px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Regulation\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003e(compared with the control group)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" style=\"width: 144px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEndometriosis\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 176px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEndometriosis+Dienogest\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width: 182px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEndometriosis+Metformin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 82px;\"\u003e\n \u003cp\u003eFold Regulation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 62px;\"\u003e\n \u003cp\u003ep-value\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 102px;\"\u003e\n \u003cp\u003eFold Regulation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003ep-value\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003eFold Regulation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003ep-value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBeta-actin\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\n \u003cp\u003e1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 62px;\"\u003e\n \u003cp\u003enan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 102px;\"\u003e\n \u003cp\u003e1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003enan\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003enan\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e*PI3K\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\n \u003cp\u003e-2.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 62px;\"\u003e\n \u003cp\u003e0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 102px;\"\u003e\n \u003cp\u003e-21.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e0.01*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 106px;\"\u003e\n \u003cp\u003e-11.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-PI3K\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\n \u003cp\u003e-4.97\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 62px;\"\u003e\n \u003cp\u003e0.0002*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 102px;\"\u003e\n \u003cp\u003e-11.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e-9.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 66px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAKT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 82px;\"\u003e\n \u003cp\u003e-2.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 62px;\"\u003e\n \u003cp\u003e0.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 102px;\"\u003e\n \u003cp\u003e2.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75px;\"\u003e\n \u003cp\u003e0.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e73.86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 76px;\"\u003e\n \u003cp\u003e0.05*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e*p\u0026lt;0.05 statistically significant difference\u003c/p\u003e\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eHematoxylin \u0026amp; Eosin Staining Findings\u003c/h2\u003e \u003cp\u003eIn the Endometriosis group, where ectopic endometrial tissue was observed, stromal cells were widespread in the tissue, and there was an accumulation of inflammatory cells and numerous vascularizations. In some areas, there was epithelial tissue consisting of small cells and structures similar to a glandular structure surrounded by cells. The collagen fibers, in the form of thick bundles forming a network with each other in the cellular area, were remarkable. It was observed that ectopic endometrial tissue was relatively smaller in the Endometriosis+Dienogest and Endometriosis+Metformin groups, and the tissue consisting of stromal and inflammatory cells remained in a very small area (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the control group of uterus tissue, endometrium, myometrium, and perimetrium layers were observed to be normal structure. In the endometriosis group, it was observed that the surface epithelium lost its continuity and was interrupted in places. It was determined that there were openings in the stroma and that the glands were separated from it. Some of the glands had lost their normal structure and were curled. It was also noted that the muscle fibres in the myometrium layer had lost their integrity and had separated, forming gaps between them and the endometrium. It was observed that the degeneration decreased and the integrity was preserved in the uterine in the Endometriosis+Dienogest group. However, there were still interruptions in the epithelium, separation of the glands from the stroma, and gaps in the stromal tissue. The myometrium and perimetrium layers were observed to be normal. The Endometriosis+Metformin group was similar to the Endometriosis+Dienogest group. It was observed that degenerative changes in the endometriosis group decreased. In this group, there were still some interruptions in the epithelium lining the lumen and separation of the glands from the stromal tissue. Myometrium and perimetrium layers were evident (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eImmunohistochemical Findings\u003c/h2\u003e \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e \u003ch2\u003eEndometriosis Lesion Immunohistochemical Findings\u003c/h2\u003e \u003cp\u003ePI3K expression was determined to be higher in the Endometriosis+Metformin group compared to the other groups, while p-PI3K was lower. AKT expression was higher in the Endometriosis+Dienogest group compared to the Endometriosis (p\u0026thinsp;=\u0026thinsp;0.004) and Endometriosis+Metformin (p\u0026thinsp;=\u0026thinsp;0.025) groups. p-AKT expression was found to be higher in the Endometriosis+Dienogest and Endometriosis+Metformin group compared to the Endometriosis group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eUterus Immunohistochemical Findings\u003c/h2\u003e \u003cp\u003ePI3K expression was higher in the control group compared to the other groups (p\u0026thinsp;=\u0026thinsp;0.004). Although the expression was higher in the Endometriosis group compared to the Endometriosis+Dienogest group (p\u0026thinsp;=\u0026thinsp;0.004), it was not significant compared to the Endometriosis+Metformin group. Expression was strong in the epithelium and some areas of the stroma in the control group. p-PI3K expression was higher in the Control, Endometriosis+Dienogest, and Endometriosis+Metformin groups compared to the Endometriosis group (p\u0026thinsp;=\u0026thinsp;0.004). The expression distribution was strong in the epithelium, glands, stroma, outer layer of the myometrium, and perimetrium in both the Endometriosis+Dienogest and Endometriosis+Metformin groups. AKT expression was higher in the Endometriosis group compared to the other groups (0.004). Expression was higher in the Endometriosis+Metformin group than in the Endometriosis+Dienogest group (p\u0026thinsp;=\u0026thinsp;0.002).\u003c/p\u003e \u003cp\u003eThe p-AKT expression of the Control and Endometriosis groups was higher than the Endometriosis+Dienogest and Endometriosis+Metformin groups (p\u0026thinsp;=\u0026thinsp;0.004). Expression was weak in the epithelium in the control group, strong in the glands, and moderate in the stroma; however, the inner layer of the myometrium was weak, the outer layer and the perimetrium were dense. In the Endometriosis group, the epithelium layer was weak, the gland and stroma were moderate, and the myometrium and perimetrium layers showed moderate reaction. In the Endometriosis+Dienogest group, the epithelium layer, stroma, and myometrium layer showed weak to moderate staining, while the glands and perimetrium layer showed intense staining. In the Endometriosis+Metformin group, a moderate reaction was observed in the epithelium, stroma, and myometrium, and a weak reaction in the glands (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn our study, the treatments provided improvement in endometriosis foci. The most significant improvement was found in the Endometriosis+Metformin group. Interestingly, the Endometriosis+Metformin group exhibited significantly higher serum E2 levels despite demonstrating the greatest lesion regression, suggesting that its anti-endometriotic activity may not be directly mediated through estrogen suppression. Dienogest and metformin increased activation of the PI3K-AKT signaling pathway in endometriosis foci. While RT-PCR findings showed significant upregulation of PI3K and p-PI3K expression in the dienogest group, immunohistochemical analyses demonstrated differential regulation of PI3K, p-PI3K, AKT, and p-AKT proteins in both treatment groups. These findings suggest that metformin and dienogest may exert their beneficial effects through distinct mechanisms involving modulation of PI3K/AKT signaling.\u003c/p\u003e \u003cp\u003eDienogest has been shown in multiple clinical trials to reduce endometriotic lesions and relieve pain effectively. Furthermore, its safety profile is favourable, characterised by the absence of androgenic effects and only minimal suppression of ovarian function (9\u0026ndash;12). In one experimental study, rats with surgically induced endometriosis received oral dienogest at doses of 0.3 or 1.0 mg/kg/day for a period of 28 days. Treatment with 0.3 mg/kg/day led to a significant reduction in the total endometrial lesion area, demonstrating an efficacy level comparable to that of 100 mg/kg/day of danazol. In contrast to the higher dose, dienogest 0.3 mg/kg/day did not result in alterations to uterine horn weight, indicating the absence of estrogenic effects at this level in rodents. The model involved the transplantation of endometrial-like tissue at two peritoneal sites (peritoneum and mesenterium). Following exposure to 0.3 mg/kg/day dienogest, the expression of vascular endothelial growth factor (VEGF) mRNA was measured in lesion tissue, and no significant differences were observed in comparison with the control group (13). Katayama et al. demonstrated that the oral administration of dienogest for a period of up to 14 days was capable of suppressing angiogenesis in an autologous rat model of endometriosis. The authors of the study hypothesised that this effect may be attributable to antiestrogenic activity or direct progestin-mediated regulation of VEGF expression (14). In our study, E2 and VEGF levels were lower in the Endometriosis+Dienogest group than in the untreated Endometriosis group, although the differences were not statistically significant. This suggests that the beneficial effects of dienogest may be related to local anti-proliferative and anti-inflammatory actions rather than changes in circulating E2 or VEGF levels. In contrast, metformin treatment significantly increased serum E2 levels. However, circulating estrogen levels may not reflect the local hormonal environment of endometriotic lesions. Therefore, the effects of Metformin and Dienogest are likely mediated through local mechanisms independent of systemic E2 and VEGF changes.\u003c/p\u003e \u003cp\u003eAn in vitro investigation demonstrated that metformin inhibited the inflammatory response, suppressed aromatase activation, and reduced proliferation in cultured endometriotic stromal cells under sterile conditions (15). Moreover, metformin has been demonstrated to exert an antiproliferative effect on endometrial glands (16). Furthermore, the presence of aromatase expression in endometriotic tissues was accompanied by elevated local estrogen concentrations, indicating that estradiol was produced in situ through aromatase activity (17). In addition, metformin has been reported to suppress follicle-stimulating hormone (FSH) activity and to inhibit insulin-stimulated production of progesterone and estradiol in granulosa cells (18). Consequently, the hypothesis has been put forward that metformin may play a role in the prevention of endometriosis through the suppression of both ovarian and local estrogen production. Supporting evidence comes from an in vitro study in which stromal cells derived from human ovarian endometrioma were cultured and exposed to metformin for 24 hours. Metformin has been demonstrated to exert a substantial anti-inflammatory effect, whilst concomitantly attenuating estradiol production and the proliferation of endometriotic stromal cells. In addition, metformin has been shown to inhibit cAMP-stimulated aromatase activity in endometriotic lesions, with this effect being dose-dependent (Takemura et al., 2007; Oner et al., 2010). Beyond the scope of endometriosis, metformin has been advanced as a therapeutic agent for ERα-positive breast cancer, a proposition that can be attributed to its capacity to suppress ERα expression and function (19).\u003c/p\u003e \u003cp\u003eIn another study, 27-week-old female mice were divided into control and metformin-treated groups. The metformin group was fed a diet containing 100 mg/kg metformin for six months. Compared with controls, metformin mice had higher serum estradiol (E2) levels and regular estrous cycles, while follicle-stimulating hormone (FSH) levels remained unchanged. These findings suggest that metformin may enhance ovarian function in middle-aged animals (20). In the present study, serum estradiol (E2) levels were found to be significantly elevated in the Endometriosis\u0026thinsp;+\u0026thinsp;Metformin group. Although metformin has previously been reported to suppress local estrogen production within endometriotic lesions, the present findings suggest that this effect may not necessarily translate into reduced circulating estrogen concentrations. Conversely, the reduced E2 levels observed in the Dienogest-treated group are consistent with the established endocrine actions of Dienogest described in earlier studies.\u003c/p\u003e \u003cp\u003eA study assessing oral treatments with metformin (100 mg/kg/day or 200 mg/kg/day) and letrozole (0.1 mg/kg/day) in a rat model of endometriosis found that the lowest histopathological implant scores occurred with 100 mg/kg/day metformin. The administration of metformin in this dosage resulted in the regression of endometriotic implants and a reduction in the severity of adhesions (21). In accordance with the findings of earlier research, adhesions were also observed at the sites of endometrial implants in the present study. The lowest degree of adhesion formation was observed in the Endometriosis\u0026thinsp;+\u0026thinsp;Metformin group. This reduction may be attributed to the anti-inflammatory and antiproliferative properties of metformin, which likely contribute to limiting adhesion development.\u003c/p\u003e \u003cp\u003eA study was conducted in which tissue was excised from rats during the diestrus phase in order to examine endometriosis-induced tissue. The results of the histological evaluation revealed the presence of ectopic endometrium, which was characterised by pseudostratified epithelium and vacuolated columnar cells alternating with darker cells. The surrounding stroma appeared edematous and contained dilated, congested blood vessels (22). In a study evaluating the effects of cisplatin and letrozole on surgically induced endometriosis, both treatments significantly reduced implant growth and histological scores in comparison with the untreated control group (23). Another experimental study demonstrated that ornidazole treatment limited the progression of endometriosis in a rat model through its anti-inflammatory and anti-angiogenic properties. After 14 days of therapy, lesion size was significantly reduced in the ornidazole group compared with controls. Histological evaluation with hematoxylin and eosin staining revealed endometrial atrophy, thinning of the epithelial layer, and decreased glandular tissue in treated rats. Furthermore, both microvessel density and the expression of IL-1β, IL-6, TNF-α, and VEGF within endometriotic lesions were markedly lower following ornidazole administration (24). In a study by Ozdemir et al., a morphological evaluation of endometrial implants was conducted, which revealed increased cystic changes and vascularity, accompanied by stromal inflammation, capillarization, and vascular engorgement. The endometrial epithelium and glands were more distinctly identified in these lesions. A morphometric analysis was conducted to compare the endometriotic foci in the zoledronic acid- and ranibizumab-treated groups with those in the control and other treatment groups. The results showed that the foci in the zoledronic acid- and ranibizumab-treated groups appeared morphologically reduced and embedded within the tissue. The aforementioned groups also exhibited decreased epithelial integrity, reduced gland density, stromal degeneration, diminished inflammation, lower capillary activity, and reduced vascular filling. A statistically significant difference was observed among groups in the distribution of epithelial histological scores (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), with scores in the zoledronic acid and ranibizumab groups significantly lower than those in the control, dienogest, danazol, and buserelin acetate groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (25). In the present study, ectopic endometrial tissue in the endometriosis group was characterised by abundant stromal cells, prominent inflammatory cell infiltration, and numerous blood vessels. In specific regions, the presence of epithelial tissue, characterised by low-lying cells and gland-like structures, was identified, with these structures being surrounded by stromal elements. Additionally, the presence of thick bundles of collagen fibres, forming interconnected networks within the cellular space, was also notable. In the uterine tissue, the surface epithelium of the endometriosis group exhibited loss of continuity, with interruptions in several regions. Conversely, in the Endometriosis\u0026thinsp;+\u0026thinsp;Dienogest and Endometriosis\u0026thinsp;+\u0026thinsp;Metformin groups, ectopic endometrial tissue exhibited a significant reduction, with stromal and inflammatory cells restricted to a confined area. The histological improvements observed in response to metformin treatment may be attributable to its anti-inflammatory, antiproliferative, and anti-angiogenic properties, as reported in previous studies. In a similar manner, the advantageous effects of dienogest are presumably derived from its capacity to impede endometrial proliferation and elicit regression of endometriotic lesions. Collectively, these findings suggest that both metformin and dienogest have therapeutic potential in mitigating endometriosis-associated histopathological changes and limiting disease progression.\u003c/p\u003e \u003cp\u003eIn a study investigating PI3K/AKT pathway regulation in human endometrial biopsies, ectopic tissue samples from 20 women with untreated peritoneal endometriosis and endometrial biopsies from 9 controls were analysed. The results demonstrated increased PI3K expression, reduced PTEN expression, and elevated AKT phosphorylation in ectopic endometrium compared with controls. It is noteworthy that levels of PTEN were found to be significantly lower in both ectopic tissue and eutopic endometrium in patients with minimal to mild disease compared with those with moderate to severe endometriosis. Furthermore, the p-AKT/total AKT ratio indicated higher AKT phosphorylation in endometriotic tissue from patients with minimal to mild disease. Collectively, these findings confirm dysregulation of the PI3K/AKT signaling pathway in endometriosis (7). In our study, PCR and immunohistochemical findings revealed that PI3K/AKT signalling pathway expression did not decrease in endometriosis foci within the treatment groups compared to the endometriosis group. Notably, despite reduced lesion size and histopathological improvement, there was no decrease in PI3K/AKT pathway expression. PI3K expression was highest in the control group in uterine tissue. According to the PCR results, p-PI3K expression was also highest in the control group. However, according to the immunohistochemical results, the control group was similar to the treatment groups, with the lowest expression observed in the endometriosis group. AKT expression decreased in the Endometriosis+Dienogest group; however, it was statistically significantly higher in the Endometriosis+Metformin and Endometriosis groups. These findings suggest that the histopathological improvements observed after metformin and dienogest treatment may not be solely explained by modulation of the PI3K/AKT pathway.\u003c/p\u003e \u003cp\u003eOur study has certain limitations. Only a single dose and treatment duration were examined for metformin and dienogest; therefore, dose-dependent and long-term effects could not be assessed. Functional outcomes related to fertility and pain\u0026mdash;which are clinically significant symptoms of endometriosis\u0026mdash;were not evaluated in this experimental model.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn conclusion, Metformin and Dienogest caused regression of implants in the rat model of endometriosis. However, their effects on protein expression related to proliferation and angiogenesis, as well as on serum sex hormone levels, differed. Metformin was identified as the most effective treatment at macroscopic, microscopic, and molecular levels. Further experimental and clinical studies are needed to understand the mechanism of action of Metformin better.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions:\u003c/strong\u003e N.\u0026Ccedil;., S.T., K.B., and A.C. were involved in the design of the experiments and drafted the manuscript. \u0026nbsp; N.\u0026Ccedil;., S.T., H.Ş., and G.A.M. were involved in data analysis. G.A.M., and N.\u0026Ccedil;. reviewed and edited the manuscript. All authors read and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eThis study was supported by the Pamukkale University Scientific Research Projects Coordination Unit (PAUBAP) under project number PAUHADYEK-2023-07, approved by the Pamukkale University Animal Experiments Ethics Committee.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest:\u0026nbsp;\u003c/strong\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval\u003c/strong\u003e: All experimental procedures involving animals were reviewed and approved by the Pamukkale University Animal Experiments Ethics Committee (PAUHADYEK) under approval number PAUHADYEK-2023-07.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eZondervan KT, Becker CM, Missmer SA. Endometriosis. N Engl J Med. 2020;382(13):1244-56.\u003c/li\u003e\n\u003cli\u003eKvaskoff M, Mu F, Terry KL, Harris HR, Poole EM, Farland L, et al. Endometriosis: a high-risk population for major chronic diseases? Human reproduction update. 2015;21(4):500-16.\u003c/li\u003e\n\u003cli\u003eHorne AW, Saunders PT, Abokhrais IM, Hogg L. Top ten endometriosis research priorities in the UK and Ireland. The Lancet. 2017;389(10085):2191-2.\u003c/li\u003e\n\u003cli\u003eSaraswat L, Ayansina D, Cooper K, Bhattacharya S, Horne A, Bhattacharya S. Impact of endometriosis on risk of further gynaecological surgery and cancer: a national cohort study. BJOG: An International Journal of Obstetrics \u0026amp; Gynaecology. 2018;125(1):64-72.\u003c/li\u003e\n\u003cli\u003eHe Y, Sun MM, Zhang GG, Yang J, Chen KS, Xu WW, et al. Targeting PI3K/Akt signal transduction for cancer therapy. Signal transduction and targeted therapy. 2021;6(1):425.\u003c/li\u003e\n\u003cli\u003eGentilini D, Busacca M, Di Francesco S, Vignali M, Vigano P, Di Blasio A. PI3K/Akt and ERK1/2 signalling pathways are involved in endometrial cell migration induced by 17\u0026beta;-estradiol and growth factors. Molecular human reproduction. 2007;13(5):317-22.\u003c/li\u003e\n\u003cli\u003eMadanes D, Bilotas MA, Bast\u0026oacute;n JI, Singla JJ, Meresman GF, Bara\u0026ntilde;ao RI, et al. PI3K/AKT pathway is altered in the endometriosis patient\u0026rsquo;s endometrium and presents differences according to severity stage. Gynecological Endocrinology. 2020;36(5):436-40.\u003c/li\u003e\n\u003cli\u003eOner G, Ozcelik B, Ozgun MT, Serin IS, Ozturk F, Basbug M. The effects of metformin and letrozole on endometriosis and comparison of the two treatment agents in a rat model. Human reproduction. 2010;25(4):932-7.\u003c/li\u003e\n\u003cli\u003eK\u0026ouml;hler G, Faustmann TA, Gerlinger C, Seitz C, Mueck AO. A dose-ranging study to determine the efficacy and safety of 1, 2, and 4 mg of dienogest daily for endometriosis. International Journal of Gynecology \u0026amp; Obstetrics. 2010;108(1):21-5.\u003c/li\u003e\n\u003cli\u003eSeitz C, Gerlinger C, Faustmann T, Strowitzki T. Safety of dienogest in the long-term treatment of endometriosis: a one-year, open-label, follow-up study. Fertility and Sterility. 2009;92(3):S107.\u003c/li\u003e\n\u003cli\u003eStrowitzki T, Faustmann T, Gerlinger C, Seitz C. Dienogest in the treatment of endometriosis-associated pelvic pain: a 12-week, randomized, double-blind, placebo-controlled study. European Journal of Obstetrics \u0026amp; Gynecology and Reproductive Biology. 2010;151(2):193-8.\u003c/li\u003e\n\u003cli\u003eStrowitzki T, Marr J, Gerlinger C, Faustmann T, Seitz C. Dienogest is as effective as leuprolide acetate in treating the painful symptoms of endometriosis: a 24-week, randomized, multicentre, open-label trial. Human Reproduction. 2010;25(3):633-41.\u003c/li\u003e\n\u003cli\u003eFischer O, Kaufmann-Reiche U, Moeller C, Fuhrmann U. Effects of dienogest on surgically induced endometriosis in rats after repeated oral administration. Gynecologic and obstetric investigation. 2011;72(3):145-51.\u003c/li\u003e\n\u003cli\u003eKatayama H, Katayama T, Uematsu K, Hiratsuka M, Kiyomura M, Shimizu Y, et al. Effect of dienogest administration on angiogenesis and hemodynamics in a rat endometrial autograft model. Human reproduction. 2010;25(11):2851-8.\u003c/li\u003e\n\u003cli\u003eTakemura Y, Osuga Y, Yoshino O, Hasegawa A, Hirata T, Hirota Y, et al. Metformin suppresses interleukin (IL)-1\u0026beta;-induced IL-8 production, aromatase activation, and proliferation of endometriotic stromal cells. The Journal of Clinical Endocrinology \u0026amp; Metabolism. 2007;92(8):3213-8.\u003c/li\u003e\n\u003cli\u003eLi L, Mamputu J-C, Wiernsperger N, Renier G. Signaling pathways involved in human vascular smooth muscle cell proliferation and matrix metalloproteinase-2 expression induced by leptin: inhibitory effect of metformin. Diabetes. 2005;54(7):2227-34.\u003c/li\u003e\n\u003cli\u003eAttar E, Bulun S. Aromatase and other steroidogenic genes in endometriosis: translational aspects. Human reproduction update. 2006;12(1):49-56.\u003c/li\u003e\n\u003cli\u003eMansfield R, Galea R, Brincat M, Hole D, Mason H. Metformin has direct effects on human ovarian steroidogenesis. Fertility and sterility. 2003;79(4):956-62.\u003c/li\u003e\n\u003cli\u003eKim J, Lee J, Jang SY, Kim C, Choi Y, Kim A. Anticancer effect of metformin on estrogen receptor-positive and tamoxifen-resistant breast cancer cell lines. Oncology reports. 2016;35(5):2553-60.\u003c/li\u003e\n\u003cli\u003eQin X, Du D, Chen Q, Wu M, Wu T, Wen J, et al. Metformin prevents murine ovarian aging. Aging (Albany NY). 2019;11(11):3785.\u003c/li\u003e\n\u003cli\u003eCheng J, Li C, Ying Y, Lv J, Qu X, McGowan E, et al. Metformin alleviates endometriosis and potentiates endometrial receptivity via decreasing VEGF and MMP9 and increasing leukemia inhibitor factor and HOXA10. Frontiers in pharmacology. 2022;13:750208.\u003c/li\u003e\n\u003cli\u003eElgamal DA, Othman E-ER, Ahmed SF. Ultrastructural features of eutopic endometrium in a rat model of endometriosis. Journal of Microscopy and Ultrastructure. 2016;4(1):20-7.\u003c/li\u003e\n\u003cli\u003eLi Z, Liu H, He Z, Zhang G, Lang J. Effects of cisplatin and letrozole on surgically induced endometriosis and comparison of the two medications in a rat model. European Journal of Pharmaceutical Sciences. 2016;93:132-40.\u003c/li\u003e\n\u003cli\u003eQin X, Yang H, Qiao D, Liu X, Liu L-B, Liu S-F, et al. Ornidazole Reduces the progression of endometriosis in a rat model. Gynecologic and Obstetric Investigation. 2022;87(5):316-23.\u003c/li\u003e\n\u003cli\u003eUreyen Ozdemir E, Adali E, Islimye Taskin M, Yavasoglu A, Aktug H, Oltulu F, et al. Effects of ranibizumab and zoledronic acid on endometriosis in a rat model. Archives of gynecology and obstetrics. 2022;305(1):267-74.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Endometriosis, PI3K, AKT, Metformin, Dienogest","lastPublishedDoi":"10.21203/rs.3.rs-10557372/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-10557372/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study investigates the therapeutic effects of Metformin and Dienogest on the PI3K/AKT signaling pathway in a rat model of induced endometriosis, aiming to enhance treatment strategies. Experimental groups included untreated control rats, rats with surgically induced endometriosis, and treatment groups receiving Dienogest (0.3 mg/kg/day) and Metformin (100 mg/kg/day) for 28 days. Treatments began on the 21st day post-induction and continued until sacrifice on day 49. Uterine cycle was monitored via vaginal smears, and endometrial lesion areas were reassessed pre- and post-treatment. Blood estradiol (E2), progesterone, and VEGF levels were evaluated using enzyme-linked immunosorbent assay (ELISA). The endometriotic implants and uterus were assessed using histopathological analysis. The expression of PI3K/AKT pathway proteins was evaluated in endometriosis lesions and uterine tissue using immunohistochemical and RT-PCR methods. Metformin significantly reduced lesion diameter compared with untreated and dienogest-treated groups (p\u0026thinsp;=\u0026thinsp;0.002). Serum E2 levels were significantly higher in the metformin group (p\u0026thinsp;=\u0026thinsp;0.05). Histological examination showed reduced inflammatory infiltration and tissue degeneration in both treatment groups. RT-PCR demonstrated that PI3K, p-PI3K, and AKT expression were increased in the treatment groups within the endometriosis lesions. However, PI3K and p-PI3K expression were found to be reduced in uterine tissue. AKT expression was statistically significantly higher in the metformin group (p\u0026thinsp;=\u0026thinsp;0.05). These findings confirm Metformin and Dienogest as effective agents for endometriosis regression, with molecular-level differences suggesting distinct mechanisms of action. Metformin's comprehensive effects on macroscopic, biochemical, microscopic, and molecular levels underscore its therapeutic potential. Further research is essential to unravel its mechanisms fully and explore clinical applications.\u003c/p\u003e","manuscriptTitle":"Comparative Effects of Metformin and Dienogest on PI3K/AKT Signaling in Experimental Endometriosis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-08-11 09:35:02","doi":"10.21203/rs.3.rs-10557372/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"177e2fab-5e0c-4e99-98ec-5028f41edea8","owner":[],"postedDate":"August 11th, 2026","published":true,"recentEditorialEvents":[{"type":"editorAssigned","content":"","date":"2026-08-07T10:15:58+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-08-07T10:15:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"Bratislava Medical Journal","date":"2026-08-01T08:29:30+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-08-11T09:35:02+00:00","versionOfRecord":[],"versionCreatedAt":"2026-08-11 09:35:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-10557372","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-10557372","identity":"rs-10557372","version":["v1"]},"buildId":"0kSTLZv8kDMMghZHuPxFD","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}